Novel use of cathepsin l inhibitor

Abstract

The present invention relates to novel use of a cathepsin L inhibitor Z-Phe-Tyr-CHO (C26H26N2O5), wherein the cathepsin L inhibitor Z-Phe-Tyr-CHO inhibits cancer progression and cachexia and muscle damage that are caused by cancer, and thus can be effectively used for preventing, ameliorating or treating cachexia and muscle loss, while having anticancer activity.

Description

New Uses of Cathepsin L Inhibitors

[0001] The present invention relates to new uses of cathepsin L inhibitors, and in particular to the use of cathepsin L inhibitors not only for anticancer purposes but also for the prevention and improvement of cachexia and muscle loss.

[0002]

[0003] Cachexia is a malignant wasting syndrome that causes weight loss and persistent muscle loss and is known not to improve with nutritional supplementation. Furthermore, the presence of cachexia lowers responsiveness to anticancer treatment, consequently significantly reducing survival rates. Additionally, the patient's quality of life deteriorates greatly due to the persistent deterioration of nutritional status and muscle loss.

[0004] Meanwhile, cachexia is caused by changes in neuroendocrine secretion and activity, as well as increased secretion of inflammatory cytokines, including IL-6 and TNF-α, and factors that promote catabolic metabolism. This leads to reduced appetite, alterations in metabolic processes, and the induction of fat, myocardial, and skeletal muscle loss. Among these, muscle loss is one of the defining characteristics of cachexia and shares similar mechanisms with age-related sarcopenia. Awareness of the health impact and importance of muscle loss is increasing, evidenced by the assignment of a disease code to sarcopenia in 2021. However, current treatments for sarcopenia and cachexia are based on symptomatic therapies using steroids, and there are no drugs available to actively and fundamentally treat cachexia and muscle loss. Therefore, there is an urgent need to develop therapeutic agents to prevent, improve, and treat cachexia and muscle loss occurring in cancer and other wasting diseases.

[0005]

[0006] One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer, cachexia, or muscle disease comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients.

[0007] Another object of the present invention is to provide a composition, diagnostic kit, or diagnostic device for diagnosing cancer, cachexia, or muscle disease, comprising a preparation for measuring the expression level of cathepsin L or a fragment thereof; or a gene encoding it.

[0008]

[0009] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below.

[0010]

[0011] Various embodiments of the present invention are described with reference to the drawings. In the following description, for a complete understanding of the present invention, various specific details, such as specific forms, compositions, and processes, are described. However, specific embodiments may be practiced without one or more of these specific details, or in combination with other known methods and forms. In other examples, known processes and manufacturing techniques are not described as specific details so as not to make the present invention unnecessary or obscure. Reference throughout this specification to one embodiment implies that a particular feature, form, composition, or characteristic described in association with the embodiment is included in one or more embodiments of the present invention. Accordingly, the circumstances of an embodiment expressed at various locations throughout this specification do not necessarily represent the same embodiment of the present invention. Additionally, a particular feature, form, composition, or characteristic may be combined in any suitable way in one or more embodiments.

[0012]

[0013] The inventors of the present invention confirmed the occurrence of cancer cachexia while conducting research on a lung cancer model using mouse allogeneic orthotopic transplantation and performed transcriptome analysis to discover molecular markers capable of simultaneously targeting cancer and cancer cachexia. Through transcriptome analysis of lung cancer and muscle tissues in cancer cachexia mice compared to normal mice, they searched for genes whose expression increases simultaneously in both tissues when cancer cachexia occurs. As a result, cathepsin L was found to be significantly increased and was confirmed as an effective molecular marker. The present invention was completed by confirming that administering Z-Phe-Tyr-CHO, a cathepsin L selective inhibitor, to cancer cachexia mice resulted in the improvement of cancer cachexia accompanied by an anticancer effect.

[0014]

[0015] In one embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of cancer is provided, comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients.

[0016] In this invention, “cathepsin L” is a cysteine ​​protease primarily found in lysosomes and is an enzyme that plays a crucial role in protein degradation. This enzyme regulates intracellular protein metabolism and is essential for the physiological function and survival of cells. Cathepsin L breaks down proteins into amino acids to supply nutrients required by cells and contributes to the removal of damaged proteins. Furthermore, cathepsin L is involved in various physiological processes and plays a particularly important role in immune responses. It contributes to the activation and regulation of immune cells to control inflammatory responses, supports defense mechanisms against infection, plays a significant role in the process of apoptosis, and contributes to maintaining normal cell growth and development. However, excessive activation of cathepsin L can be associated with various diseases. In this invention, increased expression of cathepsin L was observed in cancer cells.

[0017] In the present invention, a “cathepsin L inhibitor” is a substance that inhibits the activity of cathepsin L, a proteolytic enzyme, and plays an important role in various physiological and pathological processes. By inhibiting the activity of cathepsin L, an anticancer effect can be expected, which can contribute to inhibiting the growth and metastasis of cancer cells. Furthermore, cathepsin L inhibitors can help treat various inflammatory diseases by regulating inflammatory responses, and can exhibit a protective effect on nerve cells in neurodegenerative diseases. The cathepsin L inhibitor provided in the present invention is not particularly limited as long as it is a substance capable of inhibiting the expression of the cathepsin L protein or gene.

[0018] As used in the present invention, the term "pharmaceuticalally acceptable salt" refers to a salt prepared according to methods conventional in the art, and such methods are known to those skilled in the art. Specifically, the pharmaceutically acceptable salt includes, but is not limited to, salts derived from the following inorganic acids and organic acids and bases that are pharmacologically or physiologically acceptable. Examples of suitable acids may include hydrochloric acid, bromic acid, hydrobromide, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, etc. Salts derived from suitable bases may include, but are not limited to, alkali metals, e.g., sodium or potassium, and alkaline earth metals, e.g., magnesium.

[0019] In another embodiment of the present invention, the cathepsin L inhibitor is Z-Phe-Tyr-CHO(C 26 H 26 Provides a pharmaceutical composition containing N2O5.

[0020] In another embodiment of the present invention, the Z-Phe-Tyr-CHO(C 26 H 26 The present invention provides a pharmaceutical composition in which N2O5) is represented by the following chemical formula 1:

[0021] [Chemical Formula 1]

[0022]

[0023] In another embodiment of the present invention, a pharmaceutical composition is provided in which the cancer is gastric cancer, liver cancer, glioblastoma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer.

[0024]

[0025] In one embodiment of the present invention, a pharmaceutical composition for the prevention or treatment of cachexia is provided, comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients.

[0026] In the present invention, the term “cachexia” refers to a syndrome of severe systemic debilitation that appears in the terminal stages of diseases such as cancer, tuberculosis, diabetes, and AIDS. The main symptoms include a decrease in appetite and a decline in body weight and physical strength due to muscle and fat loss; the condition progresses to a state where weight decreases despite normal food intake. Cachexia can reduce the effectiveness of disease treatment and shorten the patient's life expectancy. Cancer is a disease in which abnormal cells grow uncontrollably; cancer cells divide more rapidly than normal cells and possess the characteristic of being able to infiltrate surrounding tissues or metastasize to distant sites. On the other hand, cachexia is a syndrome that appears in the terminal stages of various diseases such as cancer, tuberculosis, diabetes, and AIDS, and is caused by systemic inflammatory responses and metabolic changes. Cachexia is characterized by weight loss, decreased muscle mass, fatigue, and a general state of debilitation. Cachexia is clearly distinguished from cancer as it involves more than simple weight loss; it has a severe impact on the body's nutritional status and metabolism and can lower the patient's quality of life.

[0027] Cachexia is driven by various conditions such as altered energy balance, increased production of pro-cachexia cytokines and factors, and reduced adipose tissue. The most well-known cachexia-inducing factors are myostatin, activin, GDF15 (growth differentiation factor 15), TWEAK (tumor necrosis factor-like weak inducer of apoptosis), and inflammatory cytokines such as interferon γ (IFNγ), tumor necrosis factor α (TNFα), interleukin 1α and β (IL-1α and IL-1β), and interleukin (IL)-6. These factors induce catabolism by inducing proteolysis through the activation of the ubiquitin-proteasome system (UPS) and the autophagy-lysosome system (ALS). Under physiological conditions, serine / threonine-protein kinase (AKT) phosphorylates FoxO3, leading to cytoplasmic localization. Under cachexia conditions, AKT activity is inhibited by the influence of inflammatory cytokines or reduced levels of insulin-like growth factor 1 (IGF1). Reduced AKT activity causes the nuclear translocation of FoxO3a protein following dephosphorylation, enabling the transcription of Murf-1 and Atrogin-1. A common convergent step in regulating inflammatory cytokines is associated with nuclear factor kappa B (NF-κB), a common transcription factor that mediates cellular responses to various stimuli such as lipopolysaccharides, reactive oxygen species (ROS), and various cytokines. It has been demonstrated that TNF-α acts as an upstream element of a general pathway that upregulates inflammatory cytokines through NF-κB to produce catabolic cytokines.

[0028] In the present invention, the cachexia may include, for example, cancer cachexia, AIDS cachexia, chronic obstructive pulmonary disease cachexia, multiple sclerosis cachexia, or congestive heart failure cachexia, but is not limited thereto; specifically, it may be cancer cachexia.

[0029] In the present invention, cancers that may cause cachexia are not limited to the types thereof and may include melanoma, leukemia, lymphoma, multiple myeloma, myelodysplastic syndrome, breast cancer, head and neck cancer, esophageal cancer, stomach cancer, colorectal cancer (=colon cancer), rectal cancer, anal cancer, hepatocellular carcinoma, cholangiocarcinoma, gallbladder cancer, pancreatic cancer, lung cancer (non-small cell lung cancer, small cell lung cancer), thymic cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, ovarian cancer, cervical cancer, sarcoma, gastrointestinal stromal tumor, cancer of unknown primary site, mesothelioma, neuroendocrine tumor, skin cancer, blood cancer, etc., and more preferably may be digestive organ cancers such as stomach cancer, colorectal cancer, rectal cancer, liver cancer, gallbladder cancer, pancreatic cancer, or lung cancer or breast cancer.

[0030] In another embodiment of the present invention, the cathepsin L inhibitor is Z-Phe-Tyr-CHO(C 26 H 26 Provides a pharmaceutical composition containing N2O5.

[0031] In another embodiment of the present invention, the Z-Phe-Tyr-CHO(C 26 H 26 The present invention provides a pharmaceutical composition in which N2O5) is represented by the following chemical formula 1:

[0032] [Chemical Formula 1]

[0033]

[0034] In another embodiment of the present invention, a pharmaceutical composition is provided that improves and treats one or more symptoms of cachexia selected from the group consisting of decreased appetite, weight loss, increased fatigue, muscle weakness, decreased mobility, muscle loss, fat loss, and hematopoietic toxicity.

[0035]

[0036] In one embodiment of the present invention, a pharmaceutical composition for improving or treating muscle diseases is provided, comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients.

[0037] In another embodiment of the present invention, a pharmaceutical composition for improving or treating a muscle disease is provided, wherein the muscle disease is selected from the group comprising atony, muscular atrophy, muscular dystrophy, muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia gravis, muscle loss, and sarcopenia.

[0038] The "muscle disease" of the present invention may be one or more muscle diseases selected from the group consisting of atony, muscular atrophy, muscular dystrophy, muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia gravis, cachexia, and senile sarcopenia, but is not limited thereto. Specifically, it may be a disease such as senile muscle atrophy, muscle disease caused by cancer and chronic disease, or muscle atrophy caused by muscle disuse. More specifically, it may include senile muscle atrophy or muscular atrophy caused by cancer, muscular dystrophy, muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia gravis, cachexia, senile sarcopenia, and muscle loss. In particular, in the present invention, the muscle disease may be caused by cancer or a chronic disease.

[0039] In another embodiment of the present invention, a pharmaceutical composition for improving or treating a muscle disease is provided, wherein the muscle disease is caused by aging, decreased muscle function, muscle wasting, muscle degeneration, disused or damaged muscle.

[0040] Muscle disorders, particularly muscle loss, are one of the most significant characteristics of cachexia. This is known to be caused by increased protein catabolism and decreased protein synthesis resulting from the excessive activity of various cytokines. Cachexia involves symptoms that include muscle loss (sarcopenia), and there is significant overlap between the two. While the majority of patients with cachexia also have muscle loss (sarcopenia), not all patients exhibiting muscle loss display symptoms of cachexia. Clinically speaking, muscle loss (sarcopenia) can be described as a prodromal symptom of cachexia. Among the causes of weight loss in surgical patients, sarcopenia commonly occurs in elderly patients, whereas cachexia shows a higher disease association. In sarcopenia, basal metabolic rate decreases because fat increases while muscle decreases; however, in cachexia, both muscle and fat decrease, yet basal metabolic rate is often observed to increase.

[0041] In another embodiment of the present invention, the cathepsin L inhibitor is Z-Phe-Tyr-CHO(C 26 H 26 Provides a pharmaceutical composition containing N2O5.

[0042] In another embodiment of the present invention, the Z-Phe-Tyr-CHO(C 26 H 26 The present invention provides a pharmaceutical composition in which N2O5) is represented by the following chemical formula 1:

[0043] [Chemical Formula 1]

[0044]

[0045]

[0046] In one embodiment of the present invention, a composition for diagnosing cancer is provided, comprising cathepsin L or a fragment thereof; or a preparation for measuring the expression level of a gene encoding it.

[0047] In the present invention, the composition is intended to be applied to a biological sample isolated from a target individual, and the biological sample may include, but is not limited to, solid tissue samples, tissue culture media, liquid tissue samples, cells, or cell fragments. In addition, as non-limiting examples of biological samples, whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate It may include one or more selected from the group consisting of aspirate, organ secretions, cell, cell extract and cerebrospinal fluid, but is not limited thereto.

[0048] In the present invention, the term "desired individual" refers to an individual that has developed cancer or is highly likely to develop cancer, and may be a mammal including humans, and may be selected from the group consisting of, for example, humans, rats, mice, guinea pigs, hamsters, rabbits, monkeys, dogs, cats, cattle, horses, pigs, sheep, and goats, and preferably may be a human, but is not limited thereto.

[0049] When measuring the expression level of the marker according to the present invention from a biological sample isolated from the target individual in the present invention, the occurrence or possibility of a disease can be confirmed very quickly and easily.

[0050] In the present invention, the term "diagnosis" refers to confirming the existence or characteristics of a pathological condition. For the purposes of the present invention, the diagnosis may be to predict the possibility of the onset, growth, progression, or metastasis of cancer, or to distinguish cancer types, particularly distinguishing lung cancer or breast cancer from other cancer types.

[0051] In the present invention, the preparation for measuring the expression level of the protein may include one or more selected from the group consisting of antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to the protein, but is not limited thereto.

[0052] In the present invention, the term "antibody" refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, an antibody means an antibody that specifically binds to the protein. The antibodies of the present invention include polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. The antibodies can be easily manufactured using techniques widely known in the art. For example, polyclonal antibodies can be produced by a method widely known in the art that includes the process of injecting an antigen of the protein into an animal and collecting blood from the animal to obtain serum containing antibodies. Such polyclonal antibodies can be produced from any animal, such as a goat, rabbit, sheep, monkey, horse, pig, cattle, or dog. In addition, monoclonal antibodies may be prepared using the hybridoma method (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), which is widely known in the industry, or phage antibody library technology (see Clackson et al, Nature, 352:624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). Antibodies prepared by the above methods may be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. Furthermore, the antibodies of the present invention comprise not only a complete form having two full-length light chains and two full-length heavy chains, but also functional fragments of the antibody molecule. A functional fragment of an antibody molecule refers to a fragment that possesses at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.

[0053] In the present invention, the "oligopeptide" is a peptide composed of 2 to 20 amino acids and may include dipeptides, tripeptides, tetrapeptides, and pentapeptides, but is not limited thereto.

[0054] In the present invention, the "PNA (Peptide Nucleic Acid)" refers to an artificially synthesized polymer similar to DNA or RNA, which was first introduced in 1991 by Professors Nielsen, Egholm, Berg, and Buchardt of the University of Copenhagen, Denmark. While DNA has a phosphate-ribose sugar backbone, PNA has a repeating N-(2-aminoethyl)-glycine backbone connected by peptide bonds, which significantly increases its binding affinity and stability to DNA or RNA, and is therefore used in molecular biology, diagnostic analysis, and antisense therapy. PNA is disclosed in detail in the literature [Nielsen PE, Egholm M, Berg RH, Buchardt O (December 1991). "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide". Science 254 (5037): 1497-1500].

[0055] In the present invention, the "aptamer" is an oligonucleotide or peptide molecule, and general information regarding aptamers is disclosed in detail in the literature [Bock LC et al., Nature 355(6360):5646(1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630(2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727(1998)].

[0056] In the present invention, the agent for measuring the expression level of the gene encoding the protein may include one or more selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the gene encoding the protein, but is not limited thereto.

[0057] In the present invention, the "primer" is a fragment that recognizes a target gene sequence and includes a forward and reverse primer pair, but preferably is a primer pair that provides analysis results having specificity and sensitivity. High specificity can be conferred when the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, so that it amplifies only the target gene sequence containing the complementary primer binding site and does not induce non-specific amplification.

[0058] In the present invention, the term "probe" refers to a substance capable of specifically binding to a target substance to be detected within a sample, and means a substance capable of specifically confirming the presence of the target substance within the sample through said binding. The type of probe is not limited to substances commonly used in the industry, but preferably may be PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA, or DNA, and most preferably PNA. More specifically, the probe may be a biomaterial derived from an organism or similar, or manufactured in vitro, and may be, for example, enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, nerve cells, DNA, and RNA; DNA may include cDNA, genomic DNA, and oligonucleotides; RNA may include genomic RNA, mRNA, and oligonucleotides; and examples of proteins may include antibodies, antigens, enzymes, peptides, etc.

[0059] In the present invention, "LNA (Locked nucleic acids)" refers to nucleic acid analogs containing a 2'-O, 4'-C methylene bridge [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502]. LNA nucleosides contain common nucleic acid bases of DNA and RNA and can form base pairs according to the Watson-Crick base pairing rule. However, due to the 'locking' of the molecule caused by the methylene bridge, LNAs are unable to form an ideal shape in Watson-Crick bonding. When LNAs are included in DNA or RNA oligonucleotides, LNAs can pair more quickly with complementary nucleotide chains, thereby increasing the stability of the double helix.

[0060] In the present invention, "antisense" refers to an oligomer having a backbone between nucleotide base sequences and subunits, wherein the antisense oligomer hybridizes with a target sequence within RNA by Watson-Crick base pairing, thereby allowing the formation of a mRNA and RNA:oligomer heterodimer within the target sequence. The oligomer may have exact sequence complementarity or approximate complementarity with respect to the target sequence.

[0061] Since the information regarding the cathepsin L protein according to the present invention and the gene encoding them is known, a person skilled in the art can easily design a primer, probe, or antisense nucleotide that specifically binds to the gene encoding the protein based on this.

[0062]

[0063] According to one embodiment of the present invention, the present invention relates to a diagnostic kit for cancer comprising the diagnostic composition of the present invention.

[0064] In the present invention, the "kit" refers to a tool capable of evaluating the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. It includes not only direct labeling of a detectable substance related to a probe or antibody through reaction with a substrate, but also indirect labeling in which a label that develops color through reactivity with another directly labeled reagent is conjugated. It may include a color-developing substrate solution, washing solution, and other solutions that react with the label for color development, and may be manufactured to include the reagent components used. In the present invention, the kit may be a kit containing essential elements necessary for performing RT-PCR, and in addition to each primer pair specific to the marker gene, it may include test tubes, reaction buffer, deoxyribonucleotides (dNTPs), Taq polymerase, reverse transcriptase, DNase, RNase inhibitor, sterile water, etc. Furthermore, the kit may be a kit for detecting genes for cancer diagnosis that includes essential elements necessary for performing DNA chip analysis. A DNA chip kit comprises a substrate to which cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may comprise cDNA corresponding to a quantitative control gene or a fragment thereof. The kit of the present invention is not limited thereto, provided that it is known in the art.

[0065] In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

[0066] The kit of the present invention may further include one or more other component compositions, solutions, or devices suitable for the analysis method. For example, the kit of the present invention may further include essential elements necessary to perform a reverse transcription polymerase chain reaction. The reverse transcription polymerase chain reaction kit includes a primer pair specific to a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. It may also include a primer specific to the nucleic acid sequence of a control gene. Furthermore, the reverse transcription polymerase chain reaction kit may include a test tube or other suitable container, a reaction buffer (with varying pH and magnesium concentration), deoxyribonucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC-water, sterile water, etc.

[0067] In addition, the cancer diagnostic kit of the present invention may include essential elements necessary for performing DNA chip operations. The DNA chip kit may include a substrate to which cDNA or oligonucleotides corresponding to a gene or a fragment thereof are attached, and reagents, preparations, enzymes, etc., for producing fluorescently labeled probes. Additionally, the substrate may include cDNA or oligonucleotides corresponding to a control gene or a fragment thereof.

[0068] In addition, the cancer diagnostic kit of the present invention may include essential elements necessary for performing ELISA. The ELISA kit includes an antibody specific to the protein. The antibody is an antibody that has high specificity and affinity for the marker protein and has little cross-reactivity with other proteins, and is a monoclonal antibody, a polyclonal antibody, or a recombinant antibody. Additionally, the ELISA kit may include an antibody specific to a control protein. Furthermore, the ELISA kit may include reagents capable of detecting the bound antibody, such as a labeled secondary antibody, chromophores, an enzyme (e.g., conjugated with the antibody) and its substrate, or other substances capable of binding to the antibody.

[0069] In the present invention, the immobilizer for the antigen-antibody binding reaction may be a nitrocellulose membrane, a PVDF membrane, a well plate synthesized from polyvinyl resin or polystyrene resin, a glass slide glass, etc., but is not limited thereto.

[0070] In addition, in the present invention, the label of the secondary antibody is preferably a conventional chromogenic agent that produces a color reaction, and labels such as fluorescein and dyes such as HRP (horseradish peroxidase), alkaline phosphatase, colloid gold, FITC (poly L-lysine-fluorescein isothiocyanate), and RITC (rhodamine-B-isothiocyanate) may be used, but are not limited thereto.

[0071] In addition, in the present invention, it is preferable to use a chromogenic substrate to induce color development depending on the label that performs the color reaction, and TMB (3,3',5,5'-tetramethylbezidine), ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD (o-phenylenediamine), etc. may be used. At this time, it is more preferable that the chromogenic substrate be provided in a state dissolved in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is degraded by HRP used as a label for the secondary antibody conjugate to produce a chromogenic precipitate, and the presence or absence of the marker proteins is detected by visually confirming the degree of precipitation of this chromogenic precipitate.

[0072] In the present invention, the washing solution preferably comprises a phosphate buffer solution, NaCl, and Tween 20, and a buffer solution (PBST) composed of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20 is more preferably used. After the antigen-antibody binding reaction, a secondary antibody is reacted with the antigen-antibody conjugate, and then an appropriate amount of the washing solution is added to the immobilizer to wash 3 to 6 times. A sulfuric acid solution (H2SO4) may preferably be used as the reaction stopping solution.

[0073]

[0074] According to one embodiment of the present invention, the invention relates to a method for providing information for the diagnosis of cancer.

[0075] The method of the present invention may include the step of measuring the expression level of cathepsin L protein or a gene encoding said protein in a biological sample isolated from a target individual.

[0076] In the present invention, the object of the above invention is an object that has developed cancer or is highly likely to develop cancer, and may be a mammal including humans, and may be selected from the group consisting of, for example, humans, rats, mice, guinea pigs, hamsters, rabbits, monkeys, dogs, cats, cattle, horses, pigs, sheep, and goats, and preferably may be a human, but is not limited thereto.

[0077] In the present invention, the biological sample refers to any substance, biological body fluid, tissue, saliva, feces, microorganisms in feces, metabolites of microorganisms, or cells obtained from or derived from an individual, including whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, and nipple aspirate. It may be one or more selected from the group consisting of aspirate, bronchial aspirate, synovial fluid, joint aspirate, organ secretions, cell, cell extract and cerebrospinal fluid, but is not limited thereto.

[0078] In the present invention, the preparation for measuring the expression level of the protein may include one or more selected from the group consisting of antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to the protein.

[0079] In the present invention, the measurement of the expression level of the protein may be performed by protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Surface Enhanced Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion, Ouchteroni immunodiffusion, Rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry (LC-MS), LC-MS / MS (liquid chromatography-mass spectrometry / mass spectrometry), Western blotting, or ELISA (enzyme-linked immunosorbent assay).

[0080] In addition, in the present invention, the expression level of the protein may be measured by a multiple reaction monitoring (MRM) method.

[0081] In the present invention, for the multiple reaction monitoring method, the internal standard substance may be a synthetic peptide in which a specific amino acid constituting the target peptide is substituted with an isotope, or E. coli beta-galactosidase.

[0082] In the present invention, a preparation for measuring the expression level of a gene encoding the protein may include one or more selected from the group consisting of a primer, a probe, and an antisense nucleotide that specifically binds to the gene encoding the protein.

[0083] In the present invention, the measurement of the expression level of the gene encoding the protein may be performed by reverse transcription polymerase chain reaction (RT-PCR), competitive reverse transcription polymerase chain reaction (Competitive RT-PCR), real-time reverse transcription polymerase chain reaction (Real-time RT-PCR), RNase protection assay (RPA), Northern blotting, or a DNA chip.

[0084] In the above method of the present invention, the descriptions regarding 29 biomarkers, cancer, antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), aptamers, etc., and the descriptions regarding primers, probes, etc., overlap with those previously described, so in order to avoid excessive complexity in the specification, the detailed descriptions thereof are omitted below.

[0085] In one embodiment of the present invention, the method can predict that cancer has developed or is highly likely to develop in the target individual if the expression level of cathepsin L protein measured on a biological sample of the target individual; or the gene encoding it, is higher than that of a control group.

[0086] In the present invention, the term "control group" may refer to the expression level of the relevant biomarker protein or the gene encoding said protein in a healthy normal control group, the average or median value of the expression level of the relevant marker protein or the gene encoding said protein in a biological sample derived from a patient with pancreatic disease, or the average or median value of the expression level of the relevant marker protein or the gene encoding said protein in a biological sample derived from a cancer patient, preferably a patient with a cancer of a type other than pancreatic cancer, but is not limited thereto.

[0087] In the method of the present invention, predicting that the cancer has developed or is highly likely to develop includes not only predicting the possibility of the onset, growth, progression, or metastasis of the cancer, but also, in particular, distinguishing the disease that has developed or is suspected to have developed in the target individual from other diseases and predicting that the cancer that has developed or is suspected to have developed in the target individual is lung cancer or breast cancer.

[0088] In the present invention, the cancer may be pancreatic cancer, thyroid cancer, parathyroid cancer, stomach cancer, ovarian cancer, colorectal cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, bile duct cancer, non-Hodgkin lymphoma, Hodgkin lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, pro-anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine gland cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renopelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma or pituitary adenoma.

[0089]

[0090] In one embodiment of the present invention, a cancer diagnostic device is provided comprising: a measuring unit for measuring the expression level of cathepsin L or a fragment thereof; or a gene encoding it, with respect to a biological sample obtained from a target individual; and a detection unit for outputting the presence or absence of cancer from the level measured by the measuring unit.

[0091] In the present invention, the term "desired individual" refers to an individual that has developed cancer or is highly likely to develop cancer, and may be a mammal including humans, and may be selected from the group consisting of, for example, humans, rats, mice, guinea pigs, hamsters, rabbits, monkeys, dogs, cats, cattle, horses, pigs, sheep, and goats, and preferably may be a human, but is not limited thereto.

[0092] In the method of the present invention, descriptions regarding cathepsin L, cancer, prognosis, target individual, biological sample, control group, etc., overlap with previously described information; therefore, to avoid excessive complexity in the specification, detailed descriptions thereof are omitted below.

[0093]

[0094] The pharmaceutical composition of the present invention can be prepared by methods commonly used in the art to which the present invention belongs. The pharmaceutical composition of the present invention can be prepared as an oral or parenteral formulation, preferably as an injectable formulation which is a parenteral formulation, and can be administered via the dermal, intramuscular, peritoneal, intravenous, subcutaneous, nasal, or epidural routes.

[0095] The pharmaceutical composition of the present invention may be administered to an individual in an immunologically effective amount. The “immunologically effective amount” refers to a sufficient amount to produce a preventive effect against tuberculosis and an amount that does not cause side effects or severe or excessive immune responses. The precise dosage concentration varies depending on the specific immunogen to be administered and can be easily determined by a person skilled in the art based on factors well known in the medical field, such as the age, weight, health, gender, sensitivity of the individual to drugs, route of administration, and method of administration of the vaccinated person, and may be administered one to several times.

[0096] In addition, the composition provided in the present invention may be used as a pharmaceutical composition or a food composition, but is not limited thereto.

[0097] The "prevention" of the present invention may include, without limitation, any act that can block, suppress, or delay symptoms caused by cancer, cachexia, or muscle disease using the composition of the present invention.

[0098] The "treatment" and "improvement" of the present invention may include, without limitation, any act that enables the improvement or benefit of symptoms caused by acid-fast bacteria, particularly non-tuberculous acid-fast bacteria, by using the composition of the present invention.

[0099] In the present invention, the pharmaceutical composition may be characterized in that it is in the form of a capsule, tablet, granule, injection, ointment, powder, or beverage, and the pharmaceutical composition may be characterized in that it is intended for humans.

[0100] The pharmaceutical composition of the present invention is not limited to these, but may be formulated and used in the form of oral formulations such as powders, granules, capsules, tablets, and aqueous suspensions, as well as topical preparations, suppositories, and sterile injectable solutions, according to conventional methods. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. For oral administration, the pharmaceutically acceptable carrier may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavorings, etc. For injectable preparations, it may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc., in combination; and for topical administration, it may include bases, excipients, lubricants, preservatives, etc. The formulations of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injectables, it can be manufactured in the form of unit dosing ampoules or multiple dosing ampoules. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release formulation, etc.

[0101] Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. Additionally, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc. may be additionally included.

[0102] The routes of administration of the pharmaceutical composition according to the present invention are not limited to but include oral, intravenous, intramuscular, intra-arterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal. Oral or parenteral administration is preferred.

[0103] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intradural, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0104] The pharmaceutical composition of the present invention may vary depending on several factors including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, elimination rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art, depending on the patient's condition, body weight, degree of disease, drug form, route of administration, and duration, and may be administered at a dose of 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. The administration may be administered once a day or divided into several doses. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, coated tablet, capsule, liquid, gel, syrup, slurry, or suspension.

[0105] A food composition containing the composition of the present invention as an active ingredient can be manufactured in the form of various food products, such as beverages, chewing gum, tea, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, bread, etc. Since the food composition of the present invention is composed of plant extracts that have almost no toxicity or side effects, it can be used safely even when taken for a long period for preventive purposes.

[0106] When the composition of the present invention is included in a food composition, the amount may be added in a ratio of 0.1 to 50% of the total weight.

[0107] Here, when the above food composition is prepared in the form of a beverage, there are no special limitations other than containing the above food composition in the indicated proportions, and it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in ordinary beverages. That is, as natural carbohydrates, it may include monosaccharides such as glucose, disaccharides such as fructose, polysaccharides such as sucrose, conventional sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. Examples of the above flavoring agents include natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.).

[0108] In addition, the food composition of the present invention may contain various nutritional agents, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc.

[0109] These components may be used independently or in combination. The proportion of these additives is not particularly important, but is generally selected in the range of 0.1 to about 50 parts by weight per 100 parts by weight of the composition of the present invention.

[0110]

[0111] The cathepsin L inhibitor Z-Phe-Tyr-CHO of the present invention demonstrated an improvement in body weight, fat, and muscle loss accompanied by a significant anticancer effect in a cancer cachexia mouse model induced by orthotopic transplantation of mouse lung cancer cell lines. Furthermore, it inhibited the increase in expression of MuRF1, a muscle protein degradation factor, in the muscle tissue of cancer cachexia mice. In addition, when C2C12 myotube cells were cultured in a conditioned medium containing factors derived from lung cancer cell lines, the phenomenon of reduced myotube thickness was effectively inhibited by the cathepsin L inhibitor Z-Phe-Tyr-CHO. Based on the above animal and cell line experiments, it may be useful as a composition for the prevention, improvement, or treatment of cancer cachexia or muscle loss. Moreover, since it is accompanied by anticancer activity, it can be utilized as a pharmaceutical composition for anticancer treatment.

[0112]

[0113] Figure 1 shows the increased expression of cathepsin L in the cancer tissue and muscle tissue of cancer cachexia mice induced with lung cancer.

[0114] Figure 2 shows the lung cancer inhibitory effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO.

[0115] Figure 3 shows the inhibitory effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO on weight loss caused by cancer cachexia.

[0116] Figure 4 shows the inhibitory effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO on the reduction of adipose tissue weight caused by cancer cachexia.

[0117] Figure 5 shows the inhibitory effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO on muscle tissue weight loss caused by cancer cachexia.

[0118] Figure 6 shows the results of confirming histological changes in muscle tissue of a cancer cachexia model of the cathepsin L inhibitor Z-Phe-Tyr-CHO through hematoxylin & eosin (H&E) staining.

[0119] Figure 7 shows the results of confirming the change in expression levels of muscle atrophy indicators in muscle tissue of a cancer cachexia model of the cathepsin L inhibitor Z-Phe-Tyr-CHO through Western blot and the results of quantifying them.

[0120] Figure 8 shows the cancer-inhibiting effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO in human lung cancer cell lines.

[0121] Figure 9 confirms the migration inhibitory effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO in human breast cancer cell lines. It shows that the cathepsin L inhibitor Z-Phe-Tyr-CHO does not exhibit a relatively superior inhibitory effect on migration ability in breast cancer cell lines compared to lung cancer cell lines.

[0122] Figure 10 shows the results confirming that the cathepsin L inhibitor Z-Phe-Tyr-CHO inhibits the atrophy of C2C12 myotube cells promoted by a cancer cell-derived conditioned medium, and the results of quantifying this.

[0123]

[0124] The present invention will be described in more detail below through examples. These examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the invention.

[0125]

[0126] [Example]

[0127] [Example 1] Confirmation of the cancer treatment effect of cathepsin L inhibitor and the prevention and improvement effect of cachexia and muscle loss

[0128] [Example 1-1] Materials and Method

[0129] Six-week-old male C57BL / 6 mice (Orient Bio, Seongnam, Korea) were individually housed in a temperature-controlled room under a 12-hour light / 12-hour dark cycle and a temperature of 24±1℃. The mice were divided into three groups, with six mice in each group.

[0130] Mouse-derived lung cancer cell lines were transplanted into the lungs of the remaining two groups of mice (hereinafter referred to as “cancer cachexia mice”) via intratracheal injection, excluding the healthy normal mouse group (hereinafter referred to as the “normal group”), and body weight and feeding were observed for 7 weeks thereafter.

[0131] Starting two days after cancer cell transplantation, 200 µl of the cathepsin L inhibitor Z-Phe-Tyr-CHO (Cayman, Ann Arbor, Michigan, USA), dissolved in PBS (phosphate buffered saline), was injected intraperitoneally (ip) three times a week at a dose of 0.5 mg per kg of body weight in cancer cachexic mice (hereinafter, “treatment group”).

[0132] The control group of mice (hereinafter, “control group”) was injected with 200 µl of PBS without drug into the abdominal cavity of cancer cachexia mice.

[0133] Seven weeks after cancer cell transplantation, mice were sacrificed, and the body weight, lung weight, body weight excluding lungs, fat weight, and muscle weight of the mice were measured, and the muscles were isolated and histological and molecular biological analyses were performed.

[0134]

[0135] [Examples 1-2] Experimental Results

[0136] The experimental results of Example 1-1 are disclosed in FIGS. 1 to 7. Specifically, as disclosed in FIG. 1, the expression of cathepsin L in the cancer tissue and muscle tissue of the control group was increased at the RNA and protein levels compared to the normal group.

[0137] In addition, as shown in Figure 2, compared to the normal group, the lung weight increased due to the development of lung cancer in the control group, but decreased in the group administered cathepsin L inhibitor. This was similar when observed visually. As a result of measuring the lung cancer area through lung tissue analysis, it was confirmed that the lung cancer area decreased in the group administered cathepsin L inhibitor compared to the control group.

[0138] Meanwhile, as shown in Figure 3, compared to the normal group, the control group showed a decrease in body weight due to lung cancer-induced cachexia, but the group administered cathepsin L inhibitor showed improved body weight loss compared to the control group. When the weight of the cadavers excluding the lungs was measured, it was confirmed that the group administered cathepsin L inhibitor was increased compared to the control group.

[0139] In addition, as disclosed in Figure 4, compared to the normal group, the control group showed a decrease in the weight of epididymal white adipose tissue (eWAT) and brown adipose tissue (BAT) due to cancer cachexia, but it was confirmed that the reduction in adipose tissue weight was improved in the group administered with a cathepsin L inhibitor compared to the control group.

[0140] Meanwhile, as shown in Figure 5, compared to the normal group, the weight of the gastrocnemius (GA) and tibialis anterior (TA) muscles decreased in the control group due to cancer cachexia, but in the group administered cathepsin L inhibitor, the weight reduction of the gastrocnemius muscle was improved compared to the control group.

[0141] In addition, as shown in Figure 6, compared to the normal group, the control group showed a phenomenon where the size of the muscle tissue contracted due to cancer cachexia, but in the group administered cathepsin L inhibitor, the size of the muscle tissue improved compared to the control group.

[0142] Finally, as disclosed in Figure 7, Western blot analysis of muscle tissue confirmed that the expression of MuRF-1, a marker protein of muscle loss, was increased in the control group compared to the normal group, but decreased in the group administered with cathepsin L inhibitor compared to the control group.

[0143]

[0144] [Example 2] Confirmation of Reduction in Cancer Cell Metastasis Potential by Cathepsin L Inhibitor

[0145] [Example 2-1] Materials and Method

[0146] To compare the metastatic ability of cathepsin L inhibitors in lung cancer cells and breast cancer cells, A549 human lung cancer cells and MDA-MB-231 human breast cancer cells were purchased from ATCC (Manassas, Virginia, USA) and cultured in RPMI 1640 (Roswell Park Memorial Institute Medium) or DMEM (Dulbecco's Modified Eagle Medium) supplemented with 10% FBS (fetal bovine serum) and 1% Antibiotic-Antimycotic under 37°C and 5% CO2 conditions.

[0147] To confirm the invasive ability of cancer cells, lung cancer and breast cancer cells were treated with the cathepsin L inhibitor Z-Phe-Tyr-CHO (Cayman) at concentrations of 0.1, 1, 5, and 10 µg / ml under 5% FBS conditions using a Transwell chamber (Corning Coaster, Lowell, MA, USA). After 24 hours, invaded cells were identified by hematoxylin staining.

[0148] Finally, to evaluate the migration ability of cancer cells, a 2-well silicone insert was attached to a plate, and lung cancer cells were treated with the cathepsin L inhibitor Z-Phe-Tyr-CHO (Cayman) at a concentration of 10 μg / ml under 2% FBS conditions. After 24 hours, the migration of cancer cells was confirmed by microscopic observation.

[0149]

[0150] [Example 2-2] Experimental Results

[0151] The experimental results of Example 2-1 are disclosed in FIGS. 8 and 9. Specifically, it was confirmed that the invasive ability of cancer cells was reduced by the administration of a cathepsin L inhibitor to the A549 human lung cancer cell line, and although the cathepsin L inhibitor also reduced the invasive ability of cancer cells to the MDA-MB-231 human breast cancer cell line, the effect was not relatively superior compared to the lung cancer cell line.

[0152]

[0153] [Example 3] Confirmation of muscle atrophy with cathepsin L inhibitor

[0154] [Example 3-1] Materials and Method

[0155] C2C12 myoblasts were purchased from ATCC (Manassas, Virginia, USA) and cultured in DMEM (Dulbecco's Modified Eagle's Medium) supplemented with 10% FBS (fetal bovine serum) and 1% penicillin-streptomycin (P / S) at 37°C under 5% CO2 conditions. To induce differentiation into myotubes, when the C2C12 myoblasts became 80% confluent, the medium was replaced daily for 4 days with DMEM containing 2% horse serum and 1% P / S (hereinafter referred to as “differentiation medium”). Subsequently, to induce muscle atrophy, a medium prepared by mixing a conditioned medium obtained by culturing mouse-derived lung cancer cell lines with the differentiation medium in a 1:4 ratio (hereinafter referred to as “conditioned medium”) was applied to the differentiated myotubes. At this time, the cathepsin L inhibitor Z-Phe-Tyr-CHO was treated at a concentration of 1 μg / ml, and as a control, an equal amount of DMSO (dimethyl sulfoxide), the solvent used to dissolve the cathepsin L inhibitor, was treated. After 96 hours, muscle atrophy was measured through morphological and molecular biological analysis of myotube cells.

[0156]

[0157] [Example 3-2] Experimental Results

[0158] The experimental results of Example 3-1 are disclosed in Fig. 10. Specifically, it was confirmed that muscle atrophy, characterized by a decrease in the diameter of the myotube, occurred in myotube cells treated with conditioned medium compared to C2C12 myotube cells grown in pure differentiation medium without mixing with conditioned medium. In other words, it was confirmed that muscle atrophy was improved by treatment with the cathepsin L inhibitor Z-Phe-Tyr-CHO.

[0159] In addition, Western blot analysis was performed to confirm the expression of Atrogin1 and MuRF-1, indicators of muscle atrophy, and it was confirmed that conditioned medium-induced muscle atrophy was improved by treatment with the cathepsin L inhibitor Z-Phe-Tyr-CHO.

[0160]

[0161] From the animal experiments of the above examples, it can be seen that the cathepsin L inhibitor Z-Phe-Tyr-CHO can treat cancer and improve symptoms of cachexia such as weight loss and muscle loss. Furthermore, from myotube cell experiments, it was confirmed that this has a direct effect of improving muscle atrophy by acting directly on muscle tissue, in addition to the indirect improvement effect accompanying the cancer-inhibiting effect of the cathepsin L inhibitor Z-Phe-Tyr-CHO. Therefore, the cathepsin L inhibitor Z-Phe-Tyr-CHO can be used to prevent, improve, or treat cachexia and muscle loss while possessing anticancer activity.

[0162]

[0163] Foregoing, specific parts of the present invention have been described in detail. It is evident to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents.

[0164]

Claims

A pharmaceutical composition for the prevention or treatment of cancer comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients. In paragraph 1, The above cathepsin L inhibitor is Z-Phe-Tyr-CHO(C 26 H 26 Pharmaceutical composition containing N2O5. In paragraph 2, The above Z-Phe-Tyr-CHO(C 26 H 26 A pharmaceutical composition in which N2O5) is represented by the following chemical formula 1: [Chemical Formula 1] In paragraph 3, A pharmaceutical composition in which the above cancer is gastric cancer, liver cancer, glioblastoma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer. A pharmaceutical composition for the prevention or treatment of cachexia comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients. In paragraph 5, The above cathepsin L inhibitor is Z-Phe-Tyr-CHO(C 26 H 26 Pharmaceutical composition containing N2O5. In paragraph 6, The above Z-Phe-Tyr-CHO(C 26 H 26 A pharmaceutical composition in which N2O5) is represented by the following chemical formula 1: [Chemical Formula 1] In Paragraph 7, A pharmaceutical composition wherein the above cachexia is one or more selected from the group consisting of cancer cachexia, AIDS cachexia, chronic obstructive pulmonary disease cachexia, multiple sclerosis cachexia, or congestive heart failure cachexia. In paragraph 8, The above pharmaceutical composition is a pharmaceutical composition that improves and treats one or more symptoms of cachexia selected from the group consisting of decreased appetite, weight loss, increased fatigue, muscle weakness, decreased mobility, muscle loss, fat loss, and hematopoietic toxicity. A pharmaceutical composition for the improvement or treatment of muscle diseases comprising a cathepsin L inhibitor and a pharmaceutically acceptable salt as active ingredients. In Paragraph 10, The above cathepsin L inhibitor is Z-Phe-Tyr-CHO(C 26 H 26 Pharmaceutical composition containing N2O5. In Paragraph 11, The above Z-Phe-Tyr-CHO(C 26 H 26 A pharmaceutical composition in which N2O5) is represented by the following chemical formula 1: [Chemical Formula 1] In Paragraph 12, A pharmaceutical composition for improving or treating a muscle disease, wherein the muscle disease is selected from the group comprising atony, muscular atrophy, muscular dystrophy, muscle degeneration, muscle rigidity, amyotrophic lateral sclerosis, myasthenia gravis, muscle loss, and sarcopenia. In Paragraph 12, A pharmaceutical composition for improving or treating a muscle disease, wherein the above muscle disease is caused by aging, decreased muscle function, muscle wasting, muscle degeneration, disused or damaged muscle. A cancer diagnostic composition comprising cathepsin L or a fragment thereof; or a preparation for measuring the expression level of a gene encoding it. In paragraph 15, A diagnostic composition comprising one or more selected from the group consisting of antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to the protein, for measuring the expression level of the protein. In paragraph 15, A diagnostic composition comprising one or more selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the gene, for measuring the expression level of the gene. A diagnostic kit comprising a diagnostic composition according to any one of claims 15 to 17. In biological samples isolated from the target individual, A method for providing information for cancer diagnosis comprising the step of measuring the expression level of cathepsin L or a fragment thereof; or a gene encoding it. Regarding biological samples obtained from the target individual, Cathepsin L or a fragment thereof; or a measuring unit for measuring the expression level of a gene encoding it; and A cancer diagnostic device comprising: a detection unit that outputs the presence or absence of cancer from a level measured by the above-mentioned measuring unit. A method for the prevention or treatment of cancer by administering a cathepsin L inhibitor and a pharmaceutically acceptable salt to a subject. A method for the prevention or treatment of cachexia by administering a cathepsin L inhibitor and a pharmaceutically acceptable salt to a subject. A method for improving or treating muscle diseases by administering a cathepsin L inhibitor and a pharmaceutically acceptable salt to a subject. A cancer diagnostic method comprising a preparation for measuring the expression level of cathepsin L or a fragment thereof; or a gene encoding it.

Images