KR102228272B1 - Cancer-specific drug nanocomplex for synergistic anticancer effect - Google Patents

Abstract

The present invention relates to a tumor cell-specific self-assembled nanodrug complex exhibiting a synergistic anticancer effect, and more specifically, to a nanodrug complex which is decomposed by an enzyme present in tumor cells, so as to release an active ingredient anticancer drug and drug resistance inhibitory peptide (SMAC) at the same time. The drug complex or self-assembled nanoparticle according to the present invention is a prodrug that is decomposed by cathepsin B enzyme specifically overexpressed in tumor tissue, and releases doxorubicin and SMAC. Being specifically activated in reaction with tumor cells, the drug complex or self-assembled nanoparticle can solve side effects such as severe normal tissue damage and toxicity induced in the course of cancer prevention or treatment. In addition, the drug complex or self-assembled nanoparticle according to the present invention forms stable nanoparticles without additional polymers and carriers in an aqueous state, thus making it easier to carry out higher drug encapsulation and delivery efficiency, mass synthesis and QC than conventional polymer and carrier-based drug delivery formulations. In addition, the drug complex or self-assembled nanoparticle according to the present invention releases doxorubicin and SMAC to cancer cells, thereby exhibiting excellent anticancer efficacy by a synergistic effect, as well as overcoming drug resistance, which has been a problem in existing anticancer chemotherapy.

Description

[Cancer-specific drug nanocomplex for synergistic anticancer effect]

The present invention relates to a tumor cell-specific self-assembled nano-drug complex that exhibits an anti-cancer synergistic effect, and more specifically, it is degraded by an enzyme expressed in tumor cells to release an active ingredient, an anti-cancer drug and a cell death inducing agent. It relates to anticancer drug complexes.

Cancer is the number one cause of death in most advanced countries, including Korea, and is one of the most important diseases that humanity must overcome. Recently, amphiphilic peptides that can be specifically cleaved by cathepsin B, an enzyme that is specifically overexpressed in tumor cells, are combined with doxorubicin to form nanoparticles without additional polymers and carriers. A self-assembled nanopharmaceutical complex has been developed (Patent Document 1). This technology exists in an inactive state and is selectively activated by cathepsin B, which is overexpressed in cancer cells, and can reduce side effects that indicate damage and toxicity to normal tissues, which have been a problem of existing anticancer treatments. However, after metabolism, glycine-bound doxorubicin (hereinafter, also referred to as G-DOX) is released, because the anticancer efficacy of G-DOX is reduced by 30% or more compared to clinically used Doxorubicin. The disadvantage is that the therapeutic efficacy is limited. In addition, since G-DOX cannot solve drug resistance of tumor cells, its use is limited to resistant cancer like other anticancer drugs.

In order to treat resistant cancer, combination therapy is mainly used. This is to reduce the probability of occurrence of various resistant cell populations due to heterogeneity in tumors by using a combination of two or more drugs. Despite its many advantages, combination therapy has limitations in finding an effective combination of anticancer drugs and drugs that inhibit them, and clinical use is limited due to problems accompanying side effects due to the increase in toxicity caused by the use of multiple drugs. Has become. Therefore, there is an urgent need to develop a new anticancer drug that has excellent targeting ability for tumor cells, has low side effects on normal tissues, and has high tumor treatment effect.

Patent Document 1. Korean Registered Patent Publication No. 10-1930399

In order to solve these problems, the present inventors chemically combined SMAC and Doxorubicin, which are peptides capable of inhibiting drug resistance, through a peptide capable of specific cleavage of cathepsin B (FRRG). In the present invention, doxorubicin is released after metabolism, so that treatment efficacy is remarkably increased, and by simultaneously releasing a peptide (SMAC) capable of suppressing drug resistance, drug resistance that occurs during anticancer treatment is suppressed, thereby failing treatment and remodeling cancer. We have developed a new anticancer drug that is capable of effective anticancer chemotherapy without.

The present invention provides a pharmaceutical composition for the prevention or treatment of cancer or resistant cancer containing self-assembled nanoparticles comprising a conjugate to which a hydrophobic anticancer agent is bound to one end of a peptide represented by the following general formula 1 do.

[General Formula 1]

Ala-Val-Pro-Ile-Ala-Gln-Xaa-Arg-Arg-Gly

In the general formula 1, Xaa is any one selected from phenylalanine (Phe), valine (Val), leucine (Leu), isoleucine (Ile), and proline (Pro).

The average diameter of the self-assembled nanoparticles may be 50 to 500 nm.

The hydrophobic anticancer agents doxorubicin, cyclophosphamide, mecholrethamine, uramustine, melphalan, chlorambucil, ifosfamide ), bendamustine, carmustine, lomustine, streptozocin, busulfan, dacarbazine, temozolomide, thiotepa ( thiotepa), altretamine, duocarmycin, cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, Triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, cloparabine ( clofarabine), cystarbine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, pemetrexed, pemetrexed, pento Statin, thioguanine, camptothecin, topotecan, irinotecan, etoposide, teniposide, mitoxantrone, paclitaxel ( paclitaxel), docetaxel, izabepilone, vinblastine, vincr istine), vindesine, vinorelbine, estramustine, maytansine, DM1 (mertansine), DM4, dolastatin, auristatin E (auristatin) E), auristatin F (auristatin F), monomethyl auristatin E (monomethyl auristatin E), monomethyl auristatin F (monomethyl auristatin F) and may be any one or more selected from the group consisting of derivatives thereof. .

The resistant cancer may be a cancer having resistance to anticancer agents.

The cancer or resistant cancer is brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, epistem cell tumor, brain stem tumor, head and neck tumor, brain tumor, laryngeal cancer, oropharyngeal cancer, nasal/sinus cancer , Nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, oral cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymus cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, gastric cancer, liver cancer, gallbladder cancer, biliary tract cancer, Any one selected from the group consisting of pancreatic cancer, small intestine cancer, colon cancer, anal cancer, bladder cancer, kidney cancer, prostate cancer, testicular cancer, uterine cancer, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, squamous cell carcinoma, and skin cancer It can be more than that.

The drug complex or self-assembled nanoparticle according to the present invention is a prodrug that releases doxorubicin and SMAC, which is degraded by cathepsin B enzyme, which is specifically expressed in tumor tissue, and reacts specifically to tumor cells. Since it is activated, side effects such as severe cell damage and death induced in the process of preventing or treating cancer can be solved.

In addition, although the self-assembled nanoparticles according to the present invention do not contain a separate carrier, they exhibit the above-described effects such as tumor tissue-specific activity and cytotoxicity stabilization, and thus are not limited in clinical use at all.

In addition, it does not show toxicity to normal cells that express less cathepsin B, and has a specific preventive or therapeutic effect on tumor cells that are relatively overexpressed, thereby overcoming the side effects of damage and toxicity to normal tissues of conventional chemotherapy. I can.

In addition, the drug complex or self-assembled nanoparticle according to the present invention has very excellent cancer cell killing effect, and simultaneously releases a drug resistance inhibitory peptide to prevent a decrease in anticancer treatment efficacy due to drug resistance occurring during the treatment process. Therefore, it is possible to induce apoptosis not only against tumor cells but also resistant tumor cells that are resistant to specific drugs.

1 shows the mechanism of action of tumor cell-specific self-assembled nanoparticles (SMAC-FDOX) capable of synergistic anti-cancer effect.
Figure 2 shows a method of chemical synthesis of SMAC-FDOX of Example 1.
3 is an analysis result of the synthesized SMAC-FDOX, and shows the purity analysis result (a) using reverse phase high performance liquid chromatography (RP-HPLC) and the molecular weight analysis result (b) using malditope mass spectrometry.
Figure 4 shows the results of chemical structure analysis of SMAC-FDOX formed by combining each compound with amphiphilic peptide (AVPIAQFRRG) and doxorubicin by nuclear magnetic resonance (NMR).
5 is an in vitro analysis result of SMAC-FDOX, showing the particle distribution analysis result (a) using a particle size analyzer (DLS) and the particle shape analysis result (b) using a transmission electron microscope (TEM). .
6 is a result of an in vitro cathepsin B-specific cleavage analysis result of SMAC-FDOX, and the analysis result (a) using reverse phase high performance liquid chromatography after incubation with cathepsin B for 0, 1, 2, 3, 4 and 6 hours and After incubation with cathepsin D, E, L, MMP-9, and caspase-3 for 24 hours, the analysis result (b) using reverse phase high performance liquid chromatography is shown.
7 shows the results of molecular weight analysis (a) and the molecular weight analysis results of Doxorubicin (b) of the newly generated material cut by cathepsin B of FIG. 6(a) using a malditope mass spectrometer.
Figure 8 is a result of observing the intracellular behavior of SMAC-FDOX and Doxorubicin in various cancer cells and normal cells using a fluorescence microscope (a), the anticancer effect using the CCK assay MCF7 cancer cells (b) and H9C2 myocardial cells The results analyzed in (c) are shown.
FIG. 9 is a result of comparing the anticancer efficacy of SMAC-FDOX in MCF7 cancer cells with Doxorubicin and Doxorubicin and SMAC-treated cells. When Doxorubicin or SMAC-FDOX is treated, apoptosis inhibitory protein (cIAP1/2), As a result of analyzing the amount of cleaved caspase-3 and cleaved PARP using electrophoresis (a), MCF7 cancer cells treated with Doxorubicin, Doxorubicin and SMAC and SMAC-FDOX were stained with Annexin-V and PI. The results of the fluorescence analysis (b) using a fluorescence microscope and fluorescence analysis (c) using a flow cytometer are shown for the degree of cell death.
10 is a result of comparing the anticancer efficacy of SMAC-FDOX in Doxorubicin-resistant MCF7 cancer cells (ADR-MCF7) with Doxorubicin and Doxorubicin and SMAC-treated cells, ADR treated with Doxorubicin or SMAC-FDOX -As a result of analyzing the amount of apoptosis inhibitory protein in MCF7 using electrophoresis (a), ADR-MCF7 cancer cells treated with SMAC-FDOX and SMAC-FDOX combined with Doxorubicin, Doxorubicin and SMAC were stained with Annexin-V and PI. Thus, the result of analyzing the degree of cell death by fluorescence analysis using a fluorescence microscope (b) and anticancer effect using the CCK analysis method (c) is shown.
FIG. 11 is a result of analyzing the cancer accumulation efficiency of SMAC-FDOX, and the result of analyzing the amount of cancer accumulation using an in vivo fluorescence analyzer (IVIS) 6 hours after intravenous administration of Doxorubicin or SMAC-FDOX to the MCF7 breast cancer mouse model (a), as a result of quantitative analysis of the fluorescence intensity of the cancer tissue portion of the in vivo fluorescence analysis result of Fig. 11(a) (b), each anticancer drug (Doxorubicin) accumulated in major organs and cancer tissues ex vivo And SMAC-FDOX) was analyzed using IVIS (c), and the degree of accumulation of each anticancer agent in cancer tissue was analyzed through tissue staining (d).
12 is a result of comparing the anticancer efficacy of SMAC-FDOX in the MCF7 breast cancer mouse model with a mouse model administered with Doxorubicin and Doxorubicin and SMAC in combination, and the results of measuring the cancer size of the breast cancer mouse model over time (a), FIG. In the results of measuring the size of the cancer in 12(a), the results of analyzing the size (b), weight (c) and degree of cell death in the cancer tissue using H&E tissue staining on the 15th day are shown (d).
13 shows the results of analysis of the drug resistance inhibitory effect of SMAC-FDOX in the MCF7 breast cancer mouse model, and apoptosis inhibitory protein in cancer tissues (cIAP1/2 ) Shows the analysis results (a) using immunohistochemistry (Immunohistochemistry) and the analysis results (b) using electrophoresis.
FIG. 14 shows the results of comparing/analyzing the toxicity of SMAC-FDOX in rats with the method of concurrent administration of Doxorubicin and Doxorubicin and SMAC in mice. The results of measuring the body weight of (a), blood analysis results (b) and tissue staining results (c) 15 days after administration are shown.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention relates to a pharmaceutical composition for the prevention or treatment of cancer or resistant cancer containing self-assembled nanoparticles comprising a conjugate to which a hydrophobic anticancer agent is bound to one end of an amphiphilic peptide represented by the following general formula 1. .

[General Formula 1]

Ala-Val-Pro-Ile-Ala-Gln-Xaa-Arg-Arg-Gly

Xaa is not particularly limited as long as it is a non-polar amino acid, but is preferably selected from phenylalanine (Phe) valine (Val), leucine (Leu), isoleucine (Ile), and proline (Proline; Pro). It may be any one, most preferably phenylalanine (Phenylalanine; Phe).

The average diameter of the self-assembled nanoparticles may be 10 nm or more, and preferably 50 to 500 nm.

The self-assembled nanoparticles according to the present invention make it possible to exhibit a preventive or therapeutic effect even for resistant cancer that could not be treated with conventional anticancer agents alone. Therefore, the composition according to the present invention can serve as a revolutionary opportunity for the development of a drug delivery system and anticancer treatment.

The amphiphilic peptide represented by the general formula 1 is composed of an SMAC peptide and a peptide cleaved by a linker and cathepsin-B. The SMAC peptide is a protein secreted from the mitochondria into the cytoplasm in response to an apoptosis signal, and is known to directly interact with XIAP and other IAPs, and promotes caspase activity by decomposing binding to caspase, It is an endogenous inhibitor that effectively inhibits IAP. SMAC peptide increases the apoptosis effect of tumor cells by drugs.

Recently, various SMAC peptides or mimetics thereof have been developed to solve the problem of resistance to tumor cells, but only a form of drug delivery system that is used in combination with a simple anticancer agent or fixed to a carrier or cell-penetrating peptide is provided, even for tumor cells and normal cells. Because SMAC peptide is stable, it causes new side effects to cancer patients who require long-term treatment, such as increasing the burden on the body due to the anticancer drug used in combination, even if it is stable. In addition, in vitro, the effect of these combination administration was excellent, but there was a disadvantage that the anticancer effect of the combination administration of an anticancer agent and SMAC peptide did not differ significantly from that of the general anticancer drug administration in actual application, so it was difficult to apply it in practice. .

Accordingly, in the present invention, the SMAC peptide and the hydrophobic anticancer agent are applied specifically to tumor cells, but a stable drug delivery system was designed so as not to exhibit toxicity to normal cells or normal tissues. In the case of SMAC peptides, at least 4 to 8 amino acid residues are required to induce apoptosis in actual tumor cells, and since these are present in a mixture of hydrophobic and hydrophilic residues, conjugation with drugs is difficult. In addition, since the SMAC peptide has no intracellular permeability and no specificity for tumor cells, the present invention has been completed in consideration of various variables as described above within a range where the properties and functions of the SMAC peptide are not limited.

In the present invention, the amphiphilic peptide is represented by the general formula 1. Specifically, it is composed of a SMAC peptide containing 5 amino acids, a linker containing one amino acid, and a peptide containing 4 amino acids cleavable with cathepsin-B. Can be.

When the length of the amphiphilic peptide increases, the cell permeation efficiency rapidly decreases, nanoparticles cannot be properly formed in the solution, and when the length of the amphiphilic peptide is short, the function of each component may not be properly exhibited.

In addition, if any one of the amino acid sequences constituting the amphiphilic peptide is changed, it cannot be successfully combined with a hydrophobic anticancer agent, or the conjugate cannot form nanoparticles through self-assembly in a solution, or in tumor cells. When degraded, SMAC peptides and hydrophobic anticancer drugs may not function properly. For example, in the self-assembled nanoparticles according to the present invention, a peptide cleavable into cathepsin-B by cathepsin-B ('FR','RR' is a cleavage site in FRRG) is decomposed, and a hydrophobic anticancer agent is released. (Hereinafter, it is also referred to as an active state). Unlike conventional, the hydrophobic anticancer agent is released immediately in a state in which a part of the cleaved peptide is not bound.If the sequence of the amphiphilic peptide represented by the general formula 1 is changed, the hydrophobic anticancer agent cannot be released immediately and binds with some amino acid residues. It is released in the state (for example, G-DOX, RG-DOX, etc.), so that it cannot penetrate into the cell nucleus, and in that case, the anticancer effect of the anticancer drug is remarkably decreased (at least 30% anticancer effect is reduced compared to doxorubicin), or the cell Problems such as being released to the outside and causing side effects may occur.

Since the conjugate in which a hydrophobic anticancer agent is bound to one end of the amphiphilic peptide represented by the general formula 1 forms a spherical nanoparticle form (prodrug form) through self-assembly in a solution, for normal cells and tissues It may not exhibit toxicity at all, and it is easy to transfer and accumulate in tumor cells, and thus has remarkably excellent anticancer efficacy.

If self-assembled nanoparticles cannot be formed, and SMAC peptides and hydrophobic anticancer agents are simply mixed, they exhibit cytotoxicity to both normal cells and tumor cells, causing side effects as well as suppressing tumor cells. Various problems may occur, such as significantly lower than the self-assembled nanoparticles according to the invention.

In addition, the amphiphilic peptide represented by the general formula 1 is decomposed and activated by cathepsin-B enzyme, and the cathepsin-B enzyme is overexpressed in tumor cells and is hardly secreted to other cells. Therefore, the self-assembled nanoparticle composed of the conjugate of the amphiphilic peptide and the hydrophobic anticancer agent according to the present invention has a very low probability of showing false positives in normal cells other than tumor cells or normal tissues other than tumor tissues. If designing targeting a protease other than cathepsin B enzyme as a target, not only is it difficult to design a peptide to detect the protein, but it is also characterized at one site, so a specific prostate cancer As such, it has a therapeutic effect only for specific cancers, does not correspond to metastatic cancer, and has a problem that it may exhibit false positives as an extracellular secretion enzyme.

Therefore, the amphiphilic peptide is preferably any one selected from SEQ ID NOs: 1 to 5, and more preferably, it is more preferably represented by SEQ ID NO: 1, which has the best specificity of tumor cells.

The hydrophobic anticancer agent is not particularly limited as long as it is a hydrophobic anticancer agent, but preferably doxorubicin, cyclophosphamide, mecholrethamine, uramustine, melphalan. ), chlorambucil, ifosfamide, bendamustine, carmustine, lomustine, streptozocin, busulfan, dacarbazine (dacarbazine), temozolomide, thiotepa, altretamine, duocarmycin, cisplatin, carboplatin, nedaplatin, Oxaliplatin, satraplatin, triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine, capecitabine ( capecitabine), cladribine, clofarabine, cystarbine, floxuridine, fludarabine, gemcitabine, hydroxyurea , Methotrexate, pemetrexed, pentostatin, thioguanine, camptothecin, topotecan, irinotecan, etoposide, tenipo Seed (teniposide), mitoxantrone (mitoxantrone), paclitaxel (paclitaxel), docetaxel (docetaxel), isabepilone (izabepi lone), vinblastine, vincristine, vindesine, vinorelbine, estramustine, maytansine, DM1 (mertansine, mertansine), DM4 , Dolastatin, auristatin E, auristatin F, monomethyl auristatin E, monomethyl auristatin F and these It may be any one or more selected from the group consisting of derivatives, most preferably doxorubicin.

Since the self-assembled nanoparticles are formed into nanoparticles by self-assembly in a living body, a separate process for manufacturing in the form of nanoparticles can be omitted, and most of the problems that conventional SMAC peptides and hydrophobic anticancer drugs have (clinical Application, side effects, toxicity, etc.), and can exhibit remarkably excellent preventive or therapeutic effects for not only general cancer but also resistant cancer.

The conjugate in which a hydrophobic anticancer agent is bound to one end of the amphiphilic peptide represented by Formula 1 may be preferably represented by the following Structural Formula 1.

[Structural Formula 1]

The conjugate in which a hydrophobic anticancer agent is bound to one end of the amphiphilic peptide represented by the general formula 1 of the new structure of the present invention is a SMAC peptide (AVPIAQ; SEQ ID NO: 6), and a cathepsin-B specific peptide (XRRG); It was formed by conjugating the composed amphiphilic peptide and a hydrophobic anticancer agent. X of the peptide specific for cathepsin-B is any one selected from phenylalanine (Phe), valine (Val), leucine (Leu), isoleucine (Ile), and proline (Pro), and preferably phenylalanine (Phe ) Can be.

The amphiphilic peptide and the hydrophobic anticancer agent may be directly linked by a chemical or physical covalent bond or a non-covalent bond.

The composition according to the present invention (i) forms spherical nanoparticles by self-assembly without any kind of nanocarrier, exists in the form of a prodrug that does not show any toxicity to cells at all, so there is no side effect, and (ii) In terms of having specific activity on tumor cells, (iii) exhibiting very excellent anticancer effects even at low concentrations, (iv) exhibiting remarkably excellent anticancer effects against not only general cancer but also resistant cancer, etc., the prior art There is a difference between them.

In the present invention, the term "tumor cell" is used with the same meaning as "cancer cell", and is used interchangeably in the present invention, and specifically refers to a cell of a cancerous tissue, benign or malignant type.

In the present invention, the term "resistant cancer" is used with the same meaning as "resistant cancer", and is used interchangeably in the present invention.

In the present invention, the term "resistant cancer" refers to a cancer that exhibits extremely low sensitivity to anticancer treatment or the like, and thus the symptoms of cancer are not improved, alleviated, alleviated or treated by the treatment. The resistant cancer may be resistant to a specific anticancer agent from the beginning, or initially did not show resistance, but may occur because genes in cancer cells are mutated due to a long period of treatment and thus no longer show susceptibility to the same therapeutic agent.

In the present invention, the resistant cancer is not particularly limited thereto, but specifically, may be any cancer exhibiting resistance to anticancer drug treatment.

In the present invention, "drug-resistant cancer" is resistance to a specific drug, that is, an anticancer agent for each cancer type, that is, resistance, or when a specific anticancer agent is administered for a long time, the cancer cells acquire drug resistance to properly see the anticancer effect. It may be something that can happen if it becomes impossible.

The cancer or resistant cancer is brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, epistem cell tumor, brain stem tumor, head and neck tumor, brain tumor, laryngeal cancer, oropharyngeal cancer, nasal/sinus cancer , Nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, oral cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymus cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, gastric cancer, liver cancer, gallbladder cancer, biliary tract cancer, Any one selected from the group consisting of pancreatic cancer, small intestine cancer, colon cancer, anal cancer, bladder cancer, kidney cancer, prostate cancer, testicular cancer, uterine cancer, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, squamous cell carcinoma, and skin cancer It can be more than that.

The self-assembled nanoparticles, which are active ingredients of the present invention, each of the SMAC peptide and the hydrophobic anticancer agent have been used for the treatment of cancer in the prior art, but since they contain various problems, practical clinical application is difficult. However, the self-assembled nanoparticles of the present invention exhibit remarkably excellent effects not only against tumor cells but also resistant cancers that are resistant to specific anticancer agents/chemotherapy, so they are very stable and can selectively treat only tumor cells and resistant tumor cells. , It was confirmed that the anticancer effect, absorption and accumulation in cells, and specificity were remarkably better than when used alone.

In addition, the composition is characterized by having an effect of inhibiting the growth of cancer and resistant cancer and inducing the death of cancer and resistant cancer, and has a preventive or therapeutic effect on the cancer and resistant cancer.

In the composition of the present invention, the content of the active ingredient is not particularly limited, but may be 1 nM or more, and preferably 10 nM or more inhibits the growth of general cancer cells and resistant cancer cells, and is resistant to cancer cells as well as anticancer agents. By inducing the death of the resistant cancer cells possessed, it can exhibit an improvement, treatment or prevention effect of cancer or resistant cancer.

In the present specification, the term'including as an active ingredient' means that a self-assembled nanoparticle composed of a conjugate in which a hydrophobic anticancer agent is bound to one end of the amphiphilic peptide represented by the general formula 1 of the present invention is used to treat or prevent cancer or resistant cancer. It is meant to include an amount sufficient to achieve efficacy or activity.

In addition, in the pharmaceutical composition for the prevention or treatment of cancer or resistant cancer using the self-assembled nanoparticles as an active ingredient, the self-assembled nanoparticles are, for example, 0.001 mg/kg or more, preferably 0.1 mg/kg or more, more preferably Preferably, 10 mg/kg or more, even more preferably 100 mg/kg or more, even more preferably 250 mg/kg or more, and most preferably 0.1 g/kg or more are included. Self-assembled nanoparticles are formed as nanoparticles in the form of prodrugs in a solution, and exist in a very stable state that does not show any toxicity to cells. The upper limit of the quantity of the particles can be selected and carried out by a person skilled in the art within an appropriate range.

The pharmaceutical composition may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may be an excipient, a disintegrant, a sweetener, a binder, a coating agent, an expanding agent, a lubricant, a lubricant, or Flavoring agents and the like can be used.

The pharmaceutical composition may be formulated for administration by including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

Acceptable pharmaceutical carriers for compositions formulated as liquid solutions are sterilized and suitable for living organisms, such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and these One or more of the components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injection formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Further, it can be preferably formulated according to each disease or ingredient by using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, local injection into a tumor, etc., preferably oral It is administration.

The suitable dosage of the pharmaceutical composition varies depending on factors such as formulation method, mode of administration, patient's age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity, and is usually skilled. The qualified physician can easily determine and prescribe an effective dosage for the desired treatment or prophylaxis. According to a preferred embodiment, the daily dosage of the pharmaceutical composition is 0.01-100 mg/kg, preferably 0.5 to 10 mg/kg, and it is more preferable to administer once to several times a day.

The pharmaceutical composition is prepared in a unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the technical field to which the present invention pertains, or It can be manufactured by enclosing it in a dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention relates to the use of the pharmaceutical composition for the prevention or treatment of cancer or resistant cancer, the use of the self-assembled nanoparticles for the manufacture of a medicament for the prevention or treatment of cancer or resistant cancer, It provides a method for the prevention or treatment of cancer or resistant cancer comprising administering an acceptable amount of self-assembled nanoparticles.

In the method for preventing or treating cancer or resistant cancer of the present invention, the pharmaceutical composition itself has excellent specificity for resistant tumor cells already resistant to tumor cells and anticancer agents, so the route of administration is the target tissue. It can be administered via any conventional route as long as it can reach. The pharmaceutical composition of the present invention is not particularly limited thereto, but may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermal, oral, intranasal, pulmonary, or rectal, as desired. .

Hereinafter, the present invention will be described in more detail with reference to a preferred embodiment. However, these examples are intended to illustrate the present invention in more detail, and it will be apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereto.

Experimental material

Two kinds of peptides (Ac-AVPIAQFRRG and Ac-AVPIAQ) used in the present invention were purchased from Peptron (Daejeon, Korea). Doxorubicin was purchased from Futurechem (Seoul, Korea). 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), N-hydroxysuccinimide (NHS), deuterium oxide (D2O), NMR test tube (Norell Standard Series), proteolysis inhibitor, acetonitrile (ACN), trifluoroacetic acid (TFA ), N,N-Dimethylformamide (DMF, anhydrous), N,N-diisopropylethylamine (DIPEA), dimethyl sulfoxide (DMSO), methanol (MeOH), annexin V staining kit and hematoxylin-eosin staining solution from Sigma aldrich (USA). I bought it. Cathepsin B,D,E,L, MMP-9 and caspase-3 enzymes were purchased from R&D systems (Minnesota, USA). The TEM test plate was purchased from Electron Microscopy Sciences (USA). Cell lysing agent, Streptividin-HRP, and BCA protein quantification kit were purchased from Thermo Fisher Scientific (USA). Apoptosis inhibitory protein (cIAP1/2) antibody, cleaved caspase-3 antibody, and cleaved PARP antibody were purchased from Abcam (Hanam, Korea). RPMI 1640 medium, DMEM medium, antibiotics, and Fetal bovine serum were purchased from WELGENE (Daegu, Korea). MDA-MB231, MCF7, HT-29, 4T1, HDF, HUVEC and H9C2 were purchased from the American Type Culture Collection (USA). Balb/c mice and Balb/c nu/nu mice were purchased from Narabio (Daejeon, Korea).

Statistical analysis

In the experimental example of the present invention, a one-way ANOVA test was used to statistically analyze the significant difference between each group. If the P value was less than 0.05, it was considered statistically significant (indicated by an asterisk in the drawing).

Example 1. Synthesis of drug complex (SMAC-FDOX)

2 is a reaction scheme schematically showing a method for synthesizing a drug complex (SMAC-FDOX).

First, amphiphilic peptides (Ac-AVPIAQFRRG, 12 mg), EDC (1 mg), NHS (1.2 mg) and Doxorubicin (6 mg) containing peptides capable of inhibiting drug resistance are dissolved in 2 mL of Dimethylformamide (DMF). After that, it was reacted at room temperature for 3 hours to obtain SMAC-FDOX. The purity (a) and mass analysis (b) results of the obtained SMAC-FDOX are shown in FIG. 3. Since the drug complex (SMAC-FDOX) exists in the form of spherical nanoparticles through self-assembly in a solution, it is also referred to as self-assembled nanoparticles in the present invention.

In addition, the chemical structure analysis results of the amphiphilic peptide (AVPIAQFRRG), Doxorubicin, and SMAC-FDOX including the peptide capable of inhibiting drug resistance are shown in FIG. 4.

Experimental Example 1. Characterization of SMAC-FDOX prepared from Example 1 in vitro

The purpose of this study was to analyze the particle size, shape and characteristics of a tumor cell-specific self-assembled nano-drug complex (SMAC-FDOX) that exhibits synergistic anticancer effects.

To this end, after dispersing SMAC-FDOX (3.0 mg) in physiological saline (1 mL, Saline), the formed self-assembled nanoparticles were measured for diameter and size distribution using Zetasizer (NanoZS, Malvern, UK), The results are shown in Fig. 5A.

Figure 5a shows the result of analyzing the size distribution of the formed self-assembled nanoparticles after dissolving SMAC-FDOX in saline (Saline). As shown in FIG. 5A, it can be seen that the average diameter of the self-assembled nanoparticles formed by dissolving the drug complex of Example 1 in a solvent was 200 nm.

In order to analyze the morphology of the nanoparticles formed by the drug complex of Example 1, the results of analyzing the morphology of the nanoparticles using a transmission electron microscope (TEM) are shown in FIG. 5B.

Figure 5b shows the result of dissolving SMAC-FDOX in saline and then dispersing it in a TEM grid and analyzing it with a transmission electron microscope. As shown in FIG. 5B, it can be seen that the drug complex of Example 1 forms spherical nanoparticles in a solution.

To confirm the activation of SMAC-FDOX by cathepsin B, SMAC-FDOX (10 μmol) was added to 2-(N-morpholine)-ethanesulphonic acid (MES) buffer (200) containing cathepsin B enzyme (10 μg). μL), stored in an incubator at 37°C, and evaluated for cathepsin B specific cleavage using reverse phase high performance liquid chromatography (RT-HPLC) at 0, 1, 2, 3, 4 and 6 hours.

6A shows the drug complex (SMAC-FDOX) of Example 1 with cathepsin B and various enzymes (cathepsin D, E, L, MMP-9 and caspase-3) and 0, 1, 2, 3, 4, respectively. And after reacting for 6 hours, the result of analyzing the cleavage of the drug complex by enzymatic reaction using reverse phase high performance liquid chromatography (RP-HPLC). In the figure, the drug complex (SMAC-FDOX) of Example 1 is indicated as DDP-NPs.

6A, it was specifically confirmed that when SMAC-FDOX was reacted with cathepsin B, the SMAC-FDOX was cleaved to release doxorubicin.

6B, it was confirmed that SMAC-FDOX was not cleaved when SMAC-FDOX was reacted with cathepsins D, E, L, MMP-9, and caspase-3 other than cathepsin B for 24 hours.

6B is a mass spectrometry to confirm whether doxorubicin is the substance released after the drug complex (SMAC-FDOX) of Example 1 was specifically cleaved by cathepsin B enzyme.

To this end, after the drug complex (SMAC-FDOX) of Example 1 was specifically cleaved by cathepsin B enzyme, the released material (10 nmol) was dispersed in methanol, and then a high-resolution giant mass spectrometer (MALDI-TOF) The molecular weight was analyzed using and is shown in FIG. 7A. To confirm the released material, doxorubicin was prepared as a control material, which was also dispersed in methanol, and then the molecular weight was analyzed using a high-resolution macromass spectrometer (MALDI-TOF) and shown in FIG. 7B.

As a result of comparing and analyzing the MALDI-TOF result of Doxorubicin of FIG. 7B and the MALDI-TOF result of FIG. 7A, it was specifically confirmed that the substance released after reacting SMAC-FDOX with cathepsin B was doxorubicin (DOX).

Experimental Example 2. In vitro, cancer cell specific cytotoxicity of SMAC-FDOX prepared from Example 1

In vitro, to evaluate the intracellular behavior and toxicity of doxorubicin and SMAC-FDOX cancer cells of Example 1 and normal cells.

First, in order to evaluate the intracellular behavior, 2 x 10 ⁴ various cancer cells (MCF7, HT-29 and MDA-MB231) and normal cells (H9C2, HUVEC and HDF) were dispensed into a cell culture plate for a confocal microscope. . After stabilization for 24 hours, each of the cells was treated with doxorubicin and SMAC-FDOX and incubated in an incubator at 37° C. for 48 hours. Then, after incubation with the cell fixative for 10 minutes and additional incubation with DAPI for 20 minutes, intracellular behavior was evaluated using a confocal fluorescence microscope.

8A, it was confirmed that doxorubicin was observed in the nucleus of cells without distinction between cancer cells and normal cells, whereas SMAC-FDOX was observed in the nucleus of cells when cultured with cancer cells, and in the cytoplasm when cultured with normal cells. I did.

These results in FIG. 8A indicate that doxorubicin is a drug that exhibits cytotoxicity through a mechanism that enters the nucleus and binds to DNA, indicating cancer cell-specific toxicity of SMAC-FDOX.

In addition, in order to evaluate cytotoxicity, 5 x 10 ³ cells of MCF7 (human breast adenocarcinoma) cells per well were dispensed into a 96-well plate. After stabilization for 24 hours, each of the cells was treated with doxorubicin and SMAC-FDOX at various concentrations (0.01-100 μM) and incubated in an incubator at 37° C. for 48 hours. Then, after adding the cell culture solution containing 10 μg of CCK solution to each well, the cells were further cultured for 20 minutes, and the absorbance of the 96-well plate was 450 using VERSAmaxTM (Molecular Devices Corp., Sunnyvale, CA). Analyze at nm. As a control experiment, H9C2 (Rat BDIX heart myoblast) cells were used instead of tumor cells to proceed in the same manner as the above procedure, and cytotoxicity thereof was evaluated.

8B is a graph showing tumor cells (MCF7) after treatment with Doxorubicin or SMAC-FDOX, respectively, and then measuring the survival rate (%) of tumor cells.

Referring to FIG. 8B, it was confirmed that Doxorubicin and SMAC-FDOX exhibited similar anticancer efficacy in cancer cells.

Figure 8c is a graph showing the measurement of the survival rate (%) of tumor cells after treatment with Doxorubicin or SMAC-FDOX respectively on normal cells (H9C2).

Referring to FIG. 8C, it was confirmed that Doxorubicin exhibited apoptosis effect similar to that of cancer cells on normal cells, whereas SMAC-FDOX did not show significant cytotoxicity.

Experimental Example 3. In vitro, evaluation of the anticancer efficacy and drug resistance inhibitory efficacy of SMAC-FDOX prepared from Example 1

In vitro, the drug resistance inhibitory efficacy and apoptosis efficacy of SMAC-FDOX were compared and evaluated with doxorubicin.

^{First, 2 x 10 4} cells of MCF7 (human breast adenocarcinoma) cells per well were dispensed into a 6-well plate. After stabilization for 24 hours, each of the cells was treated with doxorubicin and SMAC-FDOX, and cultured in an incubator at 37° C. for 48 hours. Then, each well was treated with trypsin-EDTA to obtain a cell sample, and then a protein was extracted using a cell lysing agent, and the protein sample was analyzed using an electrophoresis method.

9A, it was confirmed that, when Doxorubicin was treated, cIAP1/2 was increased in the protein that induces drug resistance, whereas it was not increased in the cells treated with SMAC-FDOX. It was confirmed that the amount of caspase-3 and cleaved PARP was remarkably high in the cells treated with SMAC-FDOX.

In addition, in order to evaluate the anticancer efficacy of SMAC-FDOX, 2 x 10 ⁴ MCF7 cancer cells were dispensed into a cell culture plate for confocal microscopy. After stabilization for 24 hours, each of the cells was treated with doxorubicin, doxorubicin and SMAC combined and SMAC-FDOX, and cultured in an incubator at 37° C. for 48 hours. After culture, Annexin V and PI staining, which can evaluate the degree of cell death, were performed for 5 minutes. Then, after incubation with the cell fixative solution for 10 minutes and additional incubation with DAPI for 20 minutes, anticancer efficacy was evaluated using a confocal fluorescence microscope and a flow cytometer.

9B and 9C, it was confirmed that SMAC-FDOX exhibited apoptosis similar to doxorubicin and doxorubicin and SMAC-treated cells.

Additionally, the anticancer efficacy and drug resistance inhibitory efficacy of SMAC-FDOX were evaluated in MCF7 (ADR-MCF7) cells exhibiting resistance to doxorubicin.

The efficacy of inhibiting drug resistance using the electrophoresis method proceeded in the same manner as in FIG. 9A.

Referring to Figure 10a, ADR-MCF7 cells naturally overexpress cIAP1/2, a protein that exhibits drug resistance, significantly increased when treated with doxorubicin, but it was confirmed that the expression was suppressed when treated with SMAC-FDOX.

Anticancer efficacy using confocal fluorescence microscopy and flow cytometry was performed in the same manner as in FIGS. 9B and 9C.

Referring to Figures 10b and 10c, different from the results of Figures 9b and 9c, ADR-MCF7 significantly reduced the anticancer efficacy of doxorubicin, whereas the anticancer efficacy of SMAC-FDOX was significantly shown, and doxorubicin and SMAC combined treatment It was confirmed that it was similar to one result.

Experimental Example 4. In vivo, evaluation of tumor accumulation efficiency of SMAC-FDOX prepared from Example 1

The tumor accumulation efficiency of SMAC-FDOX in the MCF7 breast cancer mouse model was evaluated by comparison with doxorubicin.

To prepare the MCF7 breast cancer mouse model, 1 x 10 ⁷ MCF7 cells were injected subcutaneously into the left thigh of Balb/c nu/nu mice. After tumor formation, 4 mg/kg of doxorubicin and SMAC-FDOX were administered intravenously, and 6 hours later, the efficiency of accumulation in the tumor was evaluated through an in vivo fluorescence analyzer (IVIS).

In Figure 11a, compared to doxorubicin, a strong intensity of fluorescence was measured in the tumor site of the mice administered with SMAC-FDOX, and in Figure 11b, it was confirmed that about 2.73 times as much as quantitatively accumulated.

In addition, as a result of confirming the accumulation of doxorubicin and SMAC-FDOX in major organs (liver, lung, spleen, kidney and heart) and tumor tissues ex vivo, SMAC-FDOX is mainly accumulated in tumor tissues, whereas , Doxorubicin was confirmed to be distributed throughout the body.

In addition, after vascular staining (CD 31 staining) through immunohistochemistry, SMAC-FDOX was widely distributed along the vascular distribution in the tumor, whereas doxorubicin was significantly lower in the tumor. It was confirmed that the amount was accumulated.

Experimental Example 5. In vivo, evaluation of the anticancer efficacy and drug resistance inhibitory efficacy of SMAC-FDOX prepared from Example 1

The anticancer efficacy and drug resistance inhibitory efficacy of SMAC-FDOX in vivo (in vivo) were evaluated in a breast cancer mouse model made in the same manner as in Experimental Example 4.

First, in order to evaluate the anticancer efficacy of SMAC-FDOX, doxorubicin, doxorubicin and SMAC combination or SMAC-FDOX were administered intravenously once every 3 days to a breast cancer mouse model made in the same manner as in Experimental Example 4, and then once every 2 days. The size of the tumor was measured.

12A, it was confirmed that SMAC-FDOX significantly inhibited tumor size growth compared to the control group, doxorubicin, or doxorubicin and SMAC co-administered individuals.

12B and 12C, after the experiment of FIG. 12A was proceeded, the tumor was excised from the breast cancer mouse model on the 15th day, and observed and weighed. As a result, it was confirmed that SMAC-FDOX has excellent tumor growth inhibitory effect.

12D is a result of evaluating the degree of apoptosis by tissue staining the tumor excised on the 15th day, it was confirmed that the SMAC-FDOX-treated group had significantly more apoptosis than the other groups.

In addition, in order to evaluate the drug resistance inhibitory effect of SMAC-FDOX in vivo, the expression level of the drug resistance-inducing protein (cIAP1/2) in the tissue was evaluated by staining and electrophoresis of the tumor tissue extracted in FIG. 12B.

13A and 13B, the drug resistance-inducing protein was overexpressed in mice administered doxorubicin or doxorubicin and SMAC in combination, whereas the expression level of the drug resistance-inducing protein in cancer tissues extracted from the mice treated with SMAC-FDOX. It was confirmed through tissue staining (FIG. 13A) and electrophoresis (FIG. 13B) that is remarkably low.

Experimental Example 6. In vivo, evaluation of the toxicity of SMAC-FDOX prepared from Example 1

In vivo (in vivo) reduced toxicity of SMAC-FDOX was determined by using doxorubicin, doxorubicin and SMAC combined or intravenous administration of SMAC-FDOX in the same manner as in FIG. 11A, followed by weight measurement, blood analysis, and tissue staining. Evaluated.

Referring to FIG. 14A, it was confirmed that the mice administered doxorubicin or doxorubicin and SMAC continued to lose weight, whereas the mice administered SMAC-FDOX did not show a significant change in body weight.

14B, the mice administered doxorubicin or doxorubicin and SMAC co-administered significantly increased/decreased levels of Creatinine, BUN, AST, ALT, ALP, and Alb, which are indicators of liver and kidney toxicity, compared to the control group, whereas SMAC-FDOX It was confirmed that there was no significant level of change.

14C, it was confirmed that the mice administered with doxorubicin and SMAC co-administered in heart and liver tissues showed changes in tissue structure and necrosis, whereas SMAC-FDOX did not show any changes in structure and necrosis compared to the control group.

According to the results of the above experiment, the drug complex of the present invention is decomposed and activated by cathepsin B, thereby minimizing damage and toxicity to normal tissues, and simultaneously releasing doxorubicin and SMAC to exhibit excellent anticancer treatment efficacy by synergistic effect. .

Although the present invention has been described as a preferred embodiment mentioned above, it is possible to make various modifications or variations without departing from the gist and scope of the invention. In addition, the appended claims include such modifications or variations that fall within the gist of the present invention.

<Conclusion>

In order to improve the problems and limitations in which anticancer drugs were difficult to apply due to drug resistance, self-assembled nanoparticles that specifically act on tumor cells were developed. The self-assembled nanoparticles are formed by conjugating an amphiphilic peptide represented by General Formula 1 with a hydrophobic anticancer agent, and exist as spherical nanoparticles through self-assembly without any polymer or carrier in an aqueous solution. The nanoparticles have an average diameter of 50 to 500 nm, and have the advantage of being successfully cleaved and activated by cathepsin-B present in tumor cells.

That is, since the nanoparticles according to the present invention exhibit a wide range of anticancer effects even on drug-resistant tumor cells, the cost and time required for the development of new drugs in anticancer therapy can be greatly reduced.

In addition, through animal experiments, it was confirmed that it has excellent specific activity on tumor tissue, and it is successfully accumulated in tumor tissue, and FRRG peptide is cleaved by cathepsin-B, and SMAC peptide and hydrophobic anticancer agent are released from prodrug form. , It was confirmed that it works effectively on tumor tissues.

In normal cells, it remains inactive, but when introduced into tumor cells, it is activated by cathepsin-B, which is very stable and efficient. Furthermore, even if tumor cells are resistant to drugs, as long as cathepsin-B is expressed, apoptosis is primarily induced by anticancer agents, and drug resistance is secondarily blocked by SMAC peptides, so released anticancer agents By this, the death of tumor cells is induced double. Therefore, even if the tumor cells are resistant to drugs, it is possible to induce the anticancer effect of the tumor cells against the drug, and since it is possible to induce the death of the tumor cells reliably, side effects related to recurrence or resistance cancer can be greatly reduced. Bar, it is possible to overcome the limitations of conventional anticancer agents and their delivery systems.

<110> KOREA INSTITUTE OF SCIENCE AND TECHNOLOGY <120> Cancer-specific drug nanocomplex for synergistic anticancer effect <130> HPC9021 <160> 5 <170> KoPatentIn 3.0 <210> 1 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Amphiphilic Peptides <400> 1 Ala Val Pro Ile Ala Gln Phe Arg Arg Gly 1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Amphiphilic Peptides <400> 2 Ala Val Pro Ile Ala Gln Val Arg Arg Gly 1 5 10 <210> 3 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Amphiphilic Peptides <400> 3 Ala Val Pro Ile Ala Gln Leu Arg Arg Gly 1 5 10 <210> 4 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Amphiphilic Peptides <400> 4 Ala Val Pro Ile Ala Gln Ile Arg Arg Gly 1 5 10 <210> 5 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Amphiphilic Peptides <400> 5 Ala Val Pro Ile Ala Gln Pro Arg Arg Gly 1 5 10

Claims (5)

A pharmaceutical composition for the prevention or treatment of cancer or resistant cancer containing self-assembled nanoparticles comprising a conjugate to which a hydrophobic anticancer agent is bound to one end of the amphiphilic peptide represented by the following general formula 1.
[General Formula 1]
Ala-Val-Pro-Ile-Ala-Gln-Xaa-Arg-Arg-Gly
In the general formula 1, Xaa is any one selected from phenylalanine (Phe), valine (Val), leucine (Leu), isoleucine (Ile), and proline (Pro). The method of claim 1,
The average diameter of the self-assembled nanoparticles is a pharmaceutical composition for the prevention or treatment of cancer or resistant cancer, characterized in that 50 to 500 nm. The method of claim 1,
The hydrophobic anticancer agents doxorubicin, cyclophosphamide, mecholrethamine, uramustine, melphalan, chlorambucil, ifosfamide ), bendamustine, carmustine, lomustine, streptozocin, busulfan, dacarbazine, temozolomide, thiotepa ( thiotepa), altretamine, duocarmycin, cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, Triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, cloparabine ( clofarabine), cystarbine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, pemetrexed, pemetrexed, pento Statin (pentostatin), thioguanine (thioguanine), camptothecin (camptothecin), topotecan (topotecan), irinotecan (irinotecan), etoposide (etoposide), teniposide (teniposide), mitosantrone (mitoxantrone), paclitaxel ( paclitaxel), docetaxel, izabepilone, vinblastine, vincr istine), vindesine, vinorelbine, estramustine, maytansine, DM1 (mertansine), DM4, dolastatin, auristatin E (auristatin) E), auristatin F (auristatin F), monomethyl auristatin E (monomethyl auristatin E), monomethyl auristatin F (monomethyl auristatin F), characterized in that any one or more selected from the group consisting of derivatives thereof A pharmaceutical composition for the prevention or treatment of cancer or resistant cancer. The method of claim 1,
The resistant cancer is a pharmaceutical composition for preventing or treating cancer or resistant cancer, characterized in that the cancer has resistance to anticancer agents. The method of claim 1 or 4,
The cancer or resistant cancer is a brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, epistemoma, brain stem tumor, head and neck tumor, brain tumor, laryngeal cancer, oropharyngeal cancer, nasal/sinus cancer , Nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, oral cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymus cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, gastric cancer, liver cancer, gallbladder cancer, biliary tract cancer, Any one selected from the group consisting of pancreatic cancer, small intestine cancer, colon cancer, anal cancer, bladder cancer, kidney cancer, prostate cancer, testicular cancer, uterine cancer, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, squamous cell carcinoma, and skin cancer A pharmaceutical composition for the prevention or treatment of cancer or resistant cancer, characterized in that the above.

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