Conjugates comprising self assembly peptide molecule and uses thereof
Conjugates comprising self-assembled peptide molecules and uses thereof are. disclosed. The molecular self-(Self-assembly) assembly, which, is a subject of interest in the production of functional nanomaterials that can be applied to. various biomedical fields, such as drug delivery, Biogenesis, Biogenetic, and the like, has been studied in a solvent, but has been studied in a solvent, and. has been a promising research topic in the field of assembly of biomimetic cells for regulating cell functions. It is known that mitochondria function is related to cell, death, TCA including, as well as the (apoptosis), accumulation (necrosis) of protein (necroptosis) fibers in the brain (cell death) as reported in's mitochondria, an abnormally advanced protein-rich protein is observed in the mitochondria, of (Alzheimer) a, patient (Parkinson) who, is (Huntington) a, 's disease or (Amyotrophic lateral sclerosis) an abnormally advanced protein . as reported in Aβ's mitochondria,'s (beta-sheet) conformation mitochondria, which folding is associated with, accumulation of protein folding fibers in the brain, and the abnormally grown. protein has a cytotoxic effect. One aspect provides (Xaa1)n-Xaa2 a conjugate comprising a; peptide molecule having a peptide molecule, comprising the peptide molecule, a mitochondrial targeting moiety, and a fatty acid, as, set forth in any one of. SEQ ID NOs. The present invention provides a pharmaceutical composition for preventing and treating cancer, comprising the. conjugate. One aspect provides (Xaa1)n-Xaa2 a conjugate comprising a; peptide molecule having a peptide molecule, comprising the peptide molecule, a mitochondrial targeting moiety, and a fatty acid, as, set forth in any one of. SEQ ID NOs. The mitochondrial targeting moiety may be linked to a lysine amino acid of. a peptide molecule, and the fatty N- acid may be linked through a carboxyl. group of a fatty acid at a terminal end of the peptide molecule. Fiber Assembly of a synthetic material in (fibrous assembly) a, cell can be an innovative, approach that changes cellular function through interaction with cell elements leading to the exploration of mechanisms that elicit. a cytotoxic (Fibrils) effect, in addition to being applied as biomaterials, leading (organelle) to loss (integrity) of the cellular function of Cell Endothelial cells coupled to membranes such. as ribosomes or mitochondria and, thus inducing a toxicity effect. The self-(Self-assembly) assembling process is accelerated by the accumulation of peptides in the (programmed cell death) mitochondria, which is. triggered by the accumulation of peptides in the mitochondria, which is triggered by the intracellular. membrane potential of the cancer cell mitochondria, and thus the conjugate according to an embodiment is harmless to normal cells. The Xaa1 peptides may each independently, be, one, selected, from, the, group, consisting, of, a, valine, of, isoleucine- L- amino acid, C3-C10(beta-sheet) 2, 2 (beta-sheet) 2. The peptide may (Peptide) be 3 any 201 one selected from the group consisting of the, amino acid (Xaa1)n-Xaa2 residues linked through the. peptide n bond, and the peptide may be, 2 the 200, 2 amino 150, 2 acid 100, 2 number 50, 2 that 40, 2 may 30, 2 be 20, 2 represented 15, 2 by 10, 2 the 8, 2 peptide 6, 2 bond 4, 2 and 3 the peptide may be, in Xaa1 the form of SEQ, ID, NO, or, and, the, peptide, may, be, in, the, form, of, SEQ, ID, NO.71.1, to, SEQ ID NOs C3-C10, and the peptide may be in the form, of SEQ n ID 2 No. Xaa1. For example, (Xaa1)n-Xaa2 when the N- amino C- acid is linked to a peptide bond, the amino R acid may R be C3-C10 linked to a (C3-C10 peptide bond of) the amino acid, or may be. linked by, a peptide bond, and the amino (NH acid may be linked by a peptide bond, for example, in the form of, for example, at least one of the amino acid groups of SEQ Xaa2 ID NOs, (e.g. R in the amino acid group of SEQ ID NO :) (for example).2 For) example, the amino acid may be Xaa1 at least one amino C acid selected from the group consisting of SEQ ID NOs: ## STR2## STR2. # STR2# The amino Xaa2 acid may, be, an, amino, acid comprising at least one selected from the group consisting of an amine. group, an, amide group, Xaa2, a thiol group, a carboxyl group, and a, hydroxyl group, and the, amino acid may be either, an asparagine or a, cystamine comprising an amine group, or, an aspartic acid. or a cysteamine or a serine/threonine residue including a carboxyl group or a carboxyl group, or tyrosine. The hydroxy group Xaa2 (Xaa1)n-Xaa2 of the C-C- terminal carboxyl group of, the, lysine, may be substituted, with, a, group, selected, from the group consisting of, an amine group, an alkyl group, an ether group, an acyl group, an acyl, halide group, and an aldehyde group, and, the other chemical group may be. chemically bonded, and the other chemical group may be bonded to an amine group, for example. In the present specification, the cycloalkyl-3 C3-C10 bonded mutant may refer R to a cyclohexyl alanine residue in, which a R 10 cycloalkyl group is bonded to carbon (Cyclohexyl alanine; CHA) of one group. of amino acids R, and may be, (side chain) for example, a. cyclohexyl alanine residue in which a cyclohexyl group is bonded to carbon of a methyl group, such as an amino acid of an alanine, and the amino acid may refer to a side chain specific to a specific amino acid, for example. The peptide may include a beta-(beta-sheet) 2 sheath secondary structure, and the peptide may be present in the peptide or form a 2 β-(beta-sheet) 2 sheet secondary structure between two or, more amino acids present. in other peptides, but is not limited thereto, but is not limited thereto. The beta-(beta-sheet) 2 sheath secondary structure is formed through a hydrogen bond 2 between molecules that are horizontally linked as one of the. primary structures formed by interaction between amino acid sequences constituting a protein, and is formed through a hydrogen bond between molecules that are connected horizontally. It is 2 known that the beta sheet primary structure (aggregates) is related to protein flocculation or fiber formation in many human. diseases. As used herein, the (moiety) mitochondrial targeting moiety ' refers to a function of targeting a corresponding, substance TPP (triphenylphosphonium) to mitochondria inside a cell, and may be, for example, in a range of, for example, in a range of. from about. In one embodiment, a conjugate according to an embodiment may be targeted to mitochondria inside a cell due to a mitochondrial, targeting moiety included in the conjugate, and the conjugate. may enter mitochondria within the mitochondria and enter into mitochondria. In one embodiment, the conjugate may enter mitochondria due to the mitochondrial targeting, moiety, and the conjugate introduced into the mitochondria forms a (Self-assembly) self-assembled monolayer of the peptide synthesized within. the, mitochondria of the cancer cell to induce cell death in the mitochondria, and thus the conjugate or the peptide molecule may. form a self-assembly within the mitochondria of the cell. With regard to the formation of a self- assembly in the mitochondria in the cancer cell, the peptide molecule accumulates due to a large negative internal membrane potential of the cancer. cell mitochondria, and the self-assembly process may be accelerated through this. When the number of the mitochondrial targeting moieties 1 connected to (Xaa)n-Lys the peptide may be an integer selected from the number of. the amine C groups present in the, peptide represented by the peptide, the 1 number of the mitochondrial targeting (m) moieties 1 connected to the m+1 peptide may be equal to. or greater, than the number of the amino acid 1 groups present 2 in the peptide, but not limited. thereto. In an embodiment, a terminal end of N- the peptide molecule of the conjugate may be linked to a carboxyl group of the fatty acid and an amide bond . respectively. Examples of the fatty acid, may include palmitic, acid, stearic, acid, stearic acid, oleic acid, or a combination thereof. The conjugate may exhibit an amphiphilic property, and the amphiphilic property may mean a state that both polar and non-polar materials have an affinity for both polar and non.polar materials. To an embodiment of the, present invention, the conjugate according to an, embodiment of the present invention may have a hydrophobic property . a hydrophobic property, a hydrophobic property, and a hydrophilic property, and thus may exhibit amphiphilic properties as a whole. In an exemplary embodiment, the conjugate may be represented by the following Chemical Formula III: #, # STR52# 1# wherein the bonding material is represented by the following general. formula (III). The compound of formula ([I) as claimed in 1] Claim For example, the stereoisomeromer may refer to, a compound whose spatial arrangement is not an, enantiomeric isomer, and (Stereoisomer) the stereoisomeric forms thereof. are separated from the (Enantiomer) other diastereomeric forms (Diastereomer), for. example, between the right hand and the left hand, and, the stereoisomeric forms (Optical isomer) thereof may. be separated from the other diastereomeric forms, and as such, the stereoisomeric forms thereof may be - separated from the other diastereomeric forms cis-trans. Another aspect provides a pharmaceutical composition for preventing or treating cancer, comprising the conjugate or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutically acceptable salt may be in the form of. a pharmaceutically acceptable salt, and the pharmaceutically acceptable salt may include any of the addition, salts, of the acid or base and. the stereochemically isomeric forms thereof (parent compound), for example, the salt may be an inorganic acid or an, inorganic acid, and. the salt may be any salt that does not cause undesirable effects and is not particularly limited as long as it is a salt which does not cause undesirable effects. The salt includes inorganic salts and, organic salts, thereof, for, example, it, may, be, used, in, the, form, of, acid, acetic, acid, diacetic (lactic acid), acid, triacetylenic, acid, triacetylenic acid, dihydroxypyriic, acid, trisodium, gluconic, acid, triacetylenic, acid, dienoic, acid, or, methanesulfonic, acid, or, methanesulfonic, acid, p- or, methanesulfonic, acid, or methanesulfonic, acid. The number may be as follows. Also, the salt forms, of, the, salt, may, be converted into the free form by treatment with, an appropriate, base, N- or -D- acid, including salts of alkali and alkaline, earth metals, such as ammonium salts, salts, of alkali and alkaline earth metals, such as sodium salts, potassium. salts, and salts of alkali and alkaline earth metals such as calcium salts, for example. The present invention, relates to a pharmaceutical composition for preventing or treating cancer, which comprises the conjugate or a pharmaceutically acceptable salt thereof as an active ingredient for preventing or treating cancer, which is capable of inducing cell death by forming a self.assembly in mitochondria in cancer cells. The cancer for prevention or treatment of the pharmaceutical composition may be a solid cancer, or a blood cancer,and in, an embodiment of, the, present, invention, the, cancer, may, be, a, solid cancer, or, a hematological cancer, and in an embodiment, the cancer may be. selected from the group consisting of cervical cancer, liver cancer, colorectal cancer, pancreatic cancer, pancreatic cancer, lung carcinoma, lung carcinoma, lung carcinoma, lung carcinoma, and brain tumor, but is not limited thereto. The pharmaceutical composition of the present invention may be administered via any general. route so long as the " pharmaceutical " composition of the present invention may be administered via any, general route as long as the drug can reach the target tissue, and the pharmaceutical composition of. the present, invention may be, administered by any, device capable of, delivering the, active ingredient to, the target, cell, for example, in the intra,intramuscular administration, administration, intraperitoneal. administration, intranasal administration, or intra-dose administration. The pharmaceutical composition provided in the, present disclosure may be formulated, into, a, parenteral, formulation, such, as, an orally administrable formulation, a, suspension, a, lyophilized, formulation, an external, preparation for an external preparation for an external preparation for. an external preparation, a formulation for parenteral injections, or the like, for example, a formulation for an external preparation for an external preparation, or the like, as described in U.S. Pat. The pharmaceutical composition may further (include, a, pharmaceutically acceptable excipient or carrier that) can be used for formulation in addition to an active ingredient, i.e. a pharmaceutically acceptable salt thereof, in addition to an effective component, that is, one conjugate, or a pharmaceutically acceptable salt thereof. The excipients which may be used in the formulation of, the, pharmaceutical, composition, of the 1 present invention, may be one, or more selected from, the group consisting of carrier- in-vehicle, and, or, polyhydric alcohols, for example glycerol and, edible oil; for, example, glycerol and, edible, oil, for, example, glycerol and, edible, oil,β- 1 (enhancer). The pharmaceutically acceptable carrier included in the pharmaceutical composition of the present invention may, further, include, as, is, commonly, used in, the preparation, of, the, pharmaceutical composition, of the, present, invention, the, pharmaceutical composition, of, the, present, invention may further include, in addition to the. above-mentioned ingredients, the pharmaceutical composition, of, the, present, invention, may, further include a lubricant, a. humectant, a sweetening agent, Remington's Pharmaceutical Sciences (19th ed. 1995) an emulsifier, a. suspending agent, a preservative, and the like in the pharmaceutical composition of the present invention. The pharmaceutical composition may be formulated in the form of a variety of. oral dosage, forms, for, oral administration such as, for, example, a, diluent (elixirs) for oral administration, such as, for. example, a tablet for oral administration, a diluent such as, for example, one, or, more, tablets, and/a pharmaceutically acceptable carrier, such, as, for example, bicomponent or calcium/salt thereof and magnesium or calcium salt thereof and/or polyethylene glycol . in addition to the active ingredient. Also, when the, oral dosage form for oral administration is a tablet, it may be made, of one of magnesium aluminium silicate and, the like. The paste, may, include, a, binder such as/sodium carboxymethylcellulose, sodium carboxymethylcellulose, and, or, polyvinyl, pyrrolidine, and may optionally include a disintegrant, such as sodium/alginate or, a, sodium salt thereof and a disintegrant such. as sodium alginate or a sodium salt thereof, and a flavoring or sweetening agent, such as an absorbent primary colorant, a sweetening agent, and the like, as disclosed in U.S. Pat. In this case, the pharmaceutical composition may be formulated into, a solution or, suspension, in which the active ingredient is mixed in water together with a. stabilizer, or buffering agent, and, the solution or suspension may be prepared in a unit dosage form of ampoules or, vials, for example. In addition, the pharmaceutical composition, may, further, include an auxiliary agent, such as a sterilizing or/buffering agent, a preservative, a stabilizing, agent or an emulsifying agent, a salt, for, controlling osmotic pressure or a buffering agent, or other therapeutically useful substances, and may be formulated in accordance with a conventional. method for mixing or coating a mixture of two or more thereof. The content of the conjugate or the pharmaceutically acceptable salt thereof in the pharmaceutical composition may be, appropriately adjusted depending on the type of, the, intended use of the pharmaceutical composition or the like of the pharmaceutical composition, for example, %, 0.01 by %, 0.001 99 0.001 a total weight %, of 1 the 50 pharmaceutical % composition, 50. %, 0.001 90 for example, 50%, 0.1 50 by a weight ratio of between about and a weight range of from about, and may be in a range of from about, for example, from about. The therapeutically, effective amount of the conjugate or a pharmaceutically acceptable salt thereof included in the pharmaceutical composition may be adjusted according to the. amount, required for administration, to expect, the therapeutic effect, of the, disease, and, the, pharmaceutical effective, amount, may be adjusted according to the administration time and the. administration method, of the pharmaceutical composition, 0.01 for 500 mg/kg (example), by administering the therapeutically effective. amount of the pharmaceutical composition, to an amount that can/obtain the therapeutic effect of the disease or the administration. method thereof. 1 1 2 (,). The conjugate according to one aspect, has an effect of inducing loss (Self-assembly) of a complete state of the membrane and cell death through the same due to. the formation of the self-assembled monolayer of the peptide synthesized in the mitochondria of one cancer cell. The present 1 invention also provides a self-assembled peptide molecule conjugate prepared according to an embodiment of. the present invention. Also, 2 the self-assembled peptide molecule conjugates of the MALDI-TOF/TOF present invention may be prepared according to an embodiment. of the present invention. To an 3 embodiment of the present invention, the self-assembled peptide molecule conjugate prepared according to an embodiment is a. graph illustrating whether or not a self-assembled peptide molecule conjugate is self CAC(Critical aggreagtaion concentration)-assembled in a cell. To an 4 embodiment of the present invention, a self-assembled peptide molecule conjugate prepared according to an embodiment is a photograph showing an image of a scanning electron microscope (SEM) showing whether or not a self-assembled peptide molecule conjugate is self.assembled in a cell. The present 5 invention also provides a self-assembled peptide molecule conjugate prepared according to an embodiment of the present invention, and a method for producing. the same. , The present invention will be described in more. detail, through examples, but these Examples are for illustrative purposes only, and the scope of the present invention. is not limited to these Examples. Synthesis and 1. PA-Mito-FF analysis of EXAMPLES A PA-Mito-FF standard Phe-Phe-Lys(FFK) solid 3 phase synthesis method according to a standard solid phase (fluorenylmethoxycarbonyl chemistry)(Fmoc), 9- synthesis method was performed on (standard solid phase synthesis (SSPS)) the basis of fluorenylTcoccoccoccoccocciri Chemical . which is a basic unit, and synthesised as follows. In order, PA-Mito-FF to 0.25 nmol remove the 9- unreacted compound at room temperature (fluorenylmethoxycarbonyl solid-phase peptide synthesis), the produced. product was purified by means of (diisopropyl ethyl amine)(DIPEA, 500 umol) a mixture of (500 umol) trihexyl O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) (500 umol) triphenylphosphorinium bromide DMF (dimethylformamide) salt and 24 triisopropyl ethyl amine in the. presence of diisopropyl ethyl amine in the presence of DMF diisopropyl ethyl. amine in TFA/Water/the presence of (9.5:0.5:0.5) diisopropyl ethyl amine, and the resulting product. was HPLC purified through HPLC filtration, MALDI-TOF/TOF at room temperature,and then (triphenyl phosphonium, TPP) frozen under cold, conditions, (0.02 mmol) and DMF the 1- synthesized self-(1-hexyltriphenylphosphonium bromide salt) (0.04 mmol) assembled peptide molecule (0.02 mmol) conjugate was stored 12 in ice at room temperature . "PA-Mito-FF". The present 1 invention also provides a self-assembled peptide molecule conjugate prepared according to an embodiment of. the present invention. Also, 2 the self-assembled peptide molecule conjugates of the MALDI-TOF/TOF present invention may be prepared according to an embodiment. of the present invention. Self-2. assembly assay in the cells of the present invention It was confirmed 1 whether the self-assembled peptide molecular conjugate prepared in the above-mentioned Example B forms self.assembly in the mitochondria in the cell. , In, order to confirm whether or not the molecules within CAC (Critical aggregation concentration) the cell should be. self, PA-Mito-FF-assembled, the concentration of the molecules in CAC the cells. should, PA-Mito-FF be 10 much higher than that of the, 1~2 μ M cells, (pyrene) and. thus the concentration of the molecules in (Excitation wavelength) 335nm the cells was determined, to be much higher than that of, 373~378nm the cells. (I1 Phosphorescent chromaticity 384nm~396nm values occurring in (I the vicinity of, for example, in the vicinity of, and sup .sup .5. sup.)3 A ratio) of to a ratio of to of a mass of from a (I mass1 /I3 Then, the absorbance spectrum was) confirmed at a point Critical Aggregation-Concentration(CAC) where the graph. (plotting) was bent at a point where the graph (calibration plot) was bent, in contrast to the concentration value, and PA-Mito-FF the emission spectrum was confirmed and compared with a straightening line with an identical condition under the same. 3 conditions, and the results are shown in FIG. To an 3 embodiment of the present invention, the self-assembled peptide molecule conjugate prepared according to an embodiment is a. graph illustrating whether or not a self-assembled peptide molecule conjugate is self CAC(Critical aggreagtaion concentration)-assembled in a cell. It can 3 be seen that when, PA-Mito-FF the concentration of 60 uM the secondary cell within, the cell of the cell is within the range of greater than or equal to the threshold value, the method may be carried out at a concentration of. greater than or equal to. It was, PA-Mito-FF also observed by dissolving in water at a TEM (Transmission electron microscopy) (JEOL, JEM-1400) concentration in, order for a clear demonstration, and the results thereof were, 100 uM shown in Table I, and the. 4 results thereof are shown in FIG. To an 4 embodiment of the present invention, a self-assembled peptide molecule conjugate prepared according to an embodiment is a photograph showing an image of a scanning electron microscope (SEM) showing whether or not a self-assembled peptide molecule conjugate is self.assembled in a cell. It has been found out that, PA-Mito-FF as shown 4 in FIG. I), it is possible to form a self-(assembled three-fibrobladed structural body in a. cell, as shown in FIG. An anti-3. cancer cell activity assay in EXAMPLES The cell death activity of 1 the cell prepared in the above PA-Mito-FF-mentioned Example. II was confirmed to have a cell killing activity to the cancer cells. In detail, the cell viability was, measured after PA-Mito-FF cell HeLa culture (using), PC3 a (protocol) of 10% FBS, 1% penicillin/streptomycin (Life Technologies) irradiation, and the 37 °C results thereof. Alamar blue assay (Thermo Fisher Scientific) were measured after DMEM (Gibco) cell culture, and the 48 results thereof are, 5 shown in the results of the cell viability, and the results are shown in FIG. The present 5 invention also provides a self-assembled peptide molecule conjugate prepared according to an embodiment of the present invention, and a method for producing. the same. To an 5 embodiment of the present invention, the cancer PA-Mito-FF cell line has an anti-cancer activity against. the, PA-Mito-FF cancer cell line, as, shown in FIG. 18 uM I IC, and it is also possible to have an anti-cancer activity against the cell line of 65 uM, uterine cervix, as shown in SEQ ID NO.50 Values are shown. below. As a result, it was, PA-Mito-FF found that, according to one specific example of the results described above, cells were self-assembled in mitochondria in the cells and were specific to. cell line-line specific cytotoxicity. A conjugate comprising a self-assembled peptide molecule and a, use thereof are provided to, induce cell death through loss of (Self-assembly) a complete state of a membrane and cell death by forming a self-assembled monolayer of a peptide synthesized in. mitochondria of two cancer cells. A (Xaa1)n-(Xaa2) conjugate comprising a peptide; molecule having a Xaa2 peptide molecule having an amino, acid linked via a N- peptide bond and a fatty acid linked, via n a carboxyl group at a terminal Xaa1 end of, 2 the 200 peptide molecule, Xaa1 wherein the amino acid, is, an, amino, acid, having, an, amino, acid, sequence, of, SEQ, ID, NOs, and, the, amino, acid, may have a C3-C10 β-sheath secondary structure, and Xaa2 wherein the, peptide, molecule, comprises a beta-sheath secondary structure in the molecule of, SEQ ID 2 No (beta-sheet) 2, (beta-sheet) 2, (conjugate). A conjugate 1 according to any one n of the preceding claims, Xaa1, wherein the one or more substituents selected from the group consisting of SEQ ID, 2 NOs, ## STR53## wherein R is an amino, acid of, SEQ ID NO: #### STR57## wherein X is H, O - (O) ## STR52## wherein R is an alkyl group consisting of SEQ ID NO.1 . An asparagine 1 or aspartate, comprising Xaa2 an amino group, or a, cysteic acid comprising, an amide group, or a cysteic, acid comprising, an amide group, or an aspartic acid or a, serine comprising a, carboxyl group, or a serine/threonine residue, comprising a carboxyl group or a carboxyl group, or, a serine/threonine residue comprising a carboxyl group or a carboxyl group, or a serine. threonine residue comprising a carboxyl group, or a carboxyl group. What is claimed is 1: one or more of the following paragraphs of, Claims to: The number of mitochondrial targeting moieties connected 1 to the (Xaa1)n-(Xaa2) peptide is preferably an integer selected from the range of the number of amine groups present in the peptide represented by SEQ ID NOs, and is an integer selected from the number range of the amine groups present in the, peptide represented by. SEQ ID No. The conjugate 1 as claimed in any one of the preceding claims, wherein the mitochondrial targeting moiety is in the molecule of SEQ ID NO.2.2 TPP (triphenylphosphonium) and wherein the mitochondrial targeting moiety is in the. molecule of SEQ ID No.2.2 A conjugate as 1 claimed in any one of the preceding claims, wherein the peptide, and. the mitochondrial targeting moiety are linked directly or through a linker. A conjugate 1 as claimed in any one of, the preceding claims, wherein the fatty acid is palmitic acid, stearic acid, or a combination, thereof, wherein. the fatty acid is palmitic acid (stearic acid diolenoic acid or a combination thereof. A conjugate 1 as claimed in any one of the preceding claims, wherein the assembly forms a self-assembly within the mitochondria of the cells. of SEQ. A pharmaceutical 1. composition for the prevention, or treatment of cancer, comprising the conjugate as set forth in any one of the preceding claims, or a pharmaceutically acceptable salt thereof as an active ingredient. A pharmaceutical composition. according 9 to any one of the preceding claims, wherein the conjugate forms, a magnetic assembly at the mitochondria in the cancer cell to induce apoptosis. The pharmaceutical 9 composition of, the present invention, which, is, used, in, the, treatment, of, cervical, cancer, liver, cancer, pancreatic cancer . pancreatic cancer, pancreatic cancer, colorectal cancer, colorectal cancer, colorectal cancer, lung cancer, lung cancer, lung cancer, lung cancer, lung cancer, or brain tumor, as set forth in any one of Claims.