cell-penetrating peptides, conjugates comprising the peptides, and compositions comprising the conjugates

Abstract

This invention discloses cell-penetrating peptides, conjugates as active ingredients, and their applications. Specifically, this invention discloses cell-penetrating peptides having a sequence from SEQ ID NO:2 to SEQ ID NO:178, conjugates of peptides comprising fragments of a sequence from SEQ ID NO:2 to SEQ ID NO:178, or conjugates comprising peptides having at least 80% homology to the peptide sequence, and compositions comprising the conjugates.

Description

[0001] This application is a divisional application. The international application number of the original application is PCT / KR2013 / 008459, the international application date is September 17, 2013, the Chinese national application number is 201380057894.1, the entry date into the Chinese national phase is May 5, 2015, and the invention title is "Cell-penetrating peptide, conjugate containing the peptide, and composition containing the conjugate". Technical Field

[0002] This invention relates to cell-penetrating peptides derived from human telomerase reverse transcriptase (hTERT), conjugates of cell-penetrating peptides and active ingredients, and compositions comprising such conjugates. Background Technology

[0003] Although low molecular weight substances, nucleic acids, proteins, and nanoparticles possess immense potential as therapeutic agents at the molecular level, their application is limited by their inability to penetrate tissues and cell membranes. The development of systems for delivering such substances into cells has been an active research area over the past two decades. Intracellular transport of substances has become a topic in molecular treatment therapies. Low molecular weight substances, nucleic acids, or nanoparticles are transported into cells via various reagents, electroporation, or heat shock. However, finding a suitable method to deliver proteins into cells without destroying their activity and integrity remains challenging. In the 1980s, studies on the cell penetration ability of human immunodeficiency virus (HIV) revealed that the HIV-TAT protein, composed of 11 specific amino acids, plays a crucial role in intracellular transport. Therefore, in the 1990s, exploring appropriate methods for transporting proteins into cells became a key research area.

[0004] Telomeres are known to be repetitive sequences of genetic material found at the ends of chromosomes. They prevent chromosome damage or merging with other chromosomes. Telomeres shorten with each cell division, and after a certain number of cell divisions, their length becomes extremely short, to the point where the cell stops dividing and dies. On the other hand, telomere elongation is known to extend cell lifespan. As an example, cancer cells secrete an enzyme called telomerase, which prevents telomere shortening, thus leading to cancer cell proliferation.

[0005] The objective of this invention is to provide a novel peptide.

[0006] Another objective of this invention is to provide a polynucleotide encoding the novel peptide.

[0007] Another objective of this invention is to provide cell-penetrating peptides.

[0008] Another objective of the present invention is to provide useful peptides as carriers of intracellular active ingredients.

[0009] Another objective of the present invention is to provide useful peptides as carriers of intracellular active ingredients, particularly useful peptides for locally delivering active ingredients to mitochondria.

[0010] Another object of the present invention is to provide useful peptides for delivering active ingredients to mitochondria to improve, prevent or treat mitochondrial diseases or conditions.

[0011] Another objective of this invention is to provide conjugates of active ingredients and cell-penetrating peptides.

[0012] Another objective of the present invention is to provide a composition comprising an active ingredient and a cell-penetrating peptide conjugate.

[0013] Another objective of the present invention is to provide a pharmaceutical composition comprising a conjugate of an active ingredient and a cell-penetrating peptide.

[0014] Another objective of the present invention is to provide a functional cosmetic composition comprising a conjugate of an active ingredient and a cell-penetrating peptide.

[0015] Another objective of the present invention is to provide a health food composition comprising a conjugate of an active ingredient and a cell-penetrating peptide.

[0016] Another objective of the present invention is to provide a contrast agent comprising a conjugate of an active ingredient and a cell-penetrating peptide. Summary of the Invention

[0017] According to one embodiment of the present invention, the conjugate may be a conjugate of a cell-penetrating carrier peptide and an active ingredient, wherein the carrier peptide is a peptide comprising at least one amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, a peptide having at least 80% homology with the above sequence, or a fragment of the above peptide, and wherein the peptide and the fragment having at least 80% homology retain the cell-penetrating ability of any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178.

[0018] In another embodiment of the conjugate according to the invention, the fragment may consist of three or more amino acids.

[0019] In another embodiment of the conjugate according to the invention, the carrier peptide may consist of 30 or fewer amino acids.

[0020] In another embodiment of the conjugate according to the present invention, the carrier peptide may be a peptide consisting of any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178 or a peptide having at least 80% homology with the above sequences.

[0021] A contrast agent according to one embodiment of the present invention may comprise any of the above-described conjugates.

[0022] A contrast agent according to one embodiment of the present invention can be used for cell contrast imaging.

[0023] In another embodiment of the contrast agent according to the present invention, the cells may be stem cells.

[0024] The composition according to one embodiment of the present invention may comprise any of the above-described conjugates.

[0025] According to another embodiment of the composition of the present invention, the active ingredient can be used to treat or prevent diseases, and the composition can be a pharmaceutical composition.

[0026] According to another embodiment of the composition of the present invention, the active ingredient may be an active ingredient used in functional cosmetics, and the composition may be a cosmetic composition.

[0027] According to another embodiment of the composition of the present invention, the active ingredient may be an active ingredient used in functional health foods, and the composition may be a health food composition.

[0028] One embodiment of the method according to the invention is a method for delivering an active ingredient into cells, wherein the method includes applying the conjugate of any one of claims 1 to 12 to a desired object, and wherein the carrier peptide is a cell-penetrating peptide for delivering the active ingredient into cells, and wherein the peptide having at least 80% homology and fragments of the peptide retain the cell-penetrating ability of the peptide consisting of any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178.

[0029] According to another embodiment of the method of the present invention, the method can be used to locally deliver the active ingredient into the mitochondria inside the cell.

[0030] According to another embodiment of the cell-penetrating peptide of the present invention, the above-mentioned carrier peptide may be a peptide having any one or more amino acid sequences from SEQ ID NO:2 to SEQ ID NO:178.

[0031] The polynucleotides according to the present invention can encode the above-mentioned cell-penetrating peptides.

[0032] The carrier according to the present invention may contain the above-mentioned polynucleotides.

[0033] The transformed cells according to the present invention may contain the above-described carrier.

[0034] Industrial utilization

[0035] Active ingredients that are difficult to transport into cells can be easily transported into cells by using the peptides or conjugates of peptides and active ingredients disclosed in this invention. This means that the efficacy of the active ingredient can be increased and therefore the dosage of the active ingredient can be reduced. Therefore, side effects due to drug administration can be minimized and the effectiveness of treatment can be increased. In particular, when the drug is locally delivered into the mitochondria, mitochondrial-related diseases or conditions can be improved, and the effectiveness of disease prevention and treatment can be increased. In the case of cosmetics, significant effects can be produced with small amounts of active ingredients. By conjugating the peptide with a contrast material, the peptide can be used as a contrast material to monitor the process of cell transplantation or transplanted cells in cell therapy. In particular, the peptide can be practically used as a contrast material for stem cells injected into the body. Attached Figure Description

[0036] Figure 1 The number of cells taken up by cells in HeLa cells treated with FITC fusion with the peptide Pep1 of SEQ ID No:1 is depicted by FACS analysis. Control cells were treated with FITC only.

[0037] Figure 2 to Figure 29 The number of cells taken up by cells in HeLa cells treated with FITC fusion with peptides SEQ ID NO:2 to SEQ ID NO:178 is depicted by FACS analysis. Control cells were treated with FITC only.

[0038] Figure 30 to Figure 51 The number of cells taken up by Huh7 cells after treatment with FITC fusion with peptides SEQ ID NO:2 to SEQ ID NO:178 is depicted by FACS analysis. Control cells were treated with FITC only.

[0039] Figure 52 to Figure 69 The number of cells taken up by cells in human T lymphocyte cell lines (Jurkat cells) treated with FITC fusion with peptides of SEQ ID NO:2 to SEQ ID NO:178 is depicted by FACS analysis. Control cells were treated with FITC only.

[0040] Figure 70 The results of cytotoxicity and cell viability of HeLa cells treated with FITC fusion with the peptide Pep1 of SEQ ID No:1, as analyzed by FACS (flow cytometry), are presented. Control cells were treated with FITC only.

[0041] Figure 71 to Figure 86 The results of cytotoxicity and cell viability of HeLa cells treated with FITC fusion of peptides SEQ ID NO:2 to SEQ ID NO:178 are depicted by flow cytometry (FACS). Control cells were treated with FITC only. Detailed Implementation

[0042] While proteins, nucleic acids, peptides, and viruses possess immense potential as therapeutic agents, their use is limited by their inability to penetrate tissues and cell membranes. Even small molecules cannot penetrate lipid bilayers due to their structure or properties. Therefore, attempts to transport proteins, nucleic acids, peptides, or viruses into cells via electroporation, heat shock, or similar methods are difficult to make without damaging the cell membrane and maintaining the active state of these molecules. Numerous studies have shown that trans-activating transcriptional activator (TAT) proteins derived from human immunodeficiency virus (HIV) can serve as cell-penetrating peptides that can transport extremely large active substances into cells. Specifically, studies have been conducted on substances that, unlike TAT proteins that produce toxicity within cells, can transport extremely large molecules such as proteins, nucleic acids, peptides, or viruses into cells without producing any toxicity. Therefore, this invention was made by the inventors through the discovery that peptides derived from telomerase have significant efficacy as cell-penetrating peptides without significant toxicity.

[0043] Peptides are disclosed in SEQ ID NO:1 to SEQ ID NO:178 as shown in Tables 1 to 5 below. SEQ ID NO:179 is the full-length sequence of a human telomerase protein. The peptide of SEQ ID NO:1 is derived from telomerase and consists of a 16-amino acid sequence. Peptides of SEQ ID NO:2 to SEQ ID NO:77 comprise the peptide of SEQ ID NO:1. Peptides of SEQ ID NO:78 to SEQ ID NO:178 are fragments of the peptide of SEQ ID NO:1. The “name” in Table 1 below is used to distinguish peptides. In various specific embodiments of the invention, more than one peptide in SEQ ID NO:1 to SEQ ID NO:178 comprises a “synthetic peptide,” that is, a synthetic peptide of a selected region of telomerase. In this specification, the term “pep” refers herein to a peptide having the sequence of SEQ ID NO:1, or any amino sequence in SEQ ID NO:2 to SEQ ID NO:178, a peptide comprising an amino acid sequence having more than 80% homology with the above sequences, or a fragment of the above peptides.

[0044] [Table 1]

[0045]

[0046] [Table 2]

[0047] 41. pep-RIA-40 [605-627] EVRQHREARPALLTSRLRFIPKP 23 a.m. 42. pep-RIA-41 [605-628] EVRQHREARPALLTSRLRFIPKPD 24 hours 43. pep-RIA-42 [605-629] EVRQHREARPALLTSRLRFIPKPDG 25 years 44. pep-RIA-43 [605-630] EVRQHREARPALLTSRLRFIPKPDGL 26 years 45. pep-RIA-44 [605-631] EVRQHREARPALLTSRLRFIPKPDGLR 27 years 46. pep-RIA-45 [605-632] EVRQHREARPALLTSRLRFIPKPDGLRP 28 years 47. pep-RIA-46 [606-632] VRQHREARPALLTSRLRFIPKPDGLRP 27 a.m. 48. pep-RIA-47 [607-632] RQHREARPALLTSRLRFIPKPDGLRP 26 years 49. pep-RIA-48 [608-632] QHREARPALLTSRLRFIPKPDGLRP 25 years 50. pep-RIA-49 [609-632] HREARPALLTSRLRFIPKPDGLRP 24 hours 51. pep-RIA-50 [610-632] REARPALLTSRLRFIPKPDGLRP 23 a.m. 52. pep-RIA-51 [604-627] AEVRQHREARPALLTSRLRFIPKP 24 hours 53. pep-RIA-52 [604-628] AEVRQHREARPALLTSRLRFIPKPD 25 years 54. pep-RIA-53 [604-629] AEVRQHREARPALLTSRLRFIPKPDG 26 years 55. pep-RIA-54 [604-630] AEVRQHREARPALLTSRLRFIPKPDGL 27 a.m. 56. pep-RIA-55 [604-631] AEVRQHREARPALLTSRLRFIPKPDGLR 28 years 57. pep-RIA-56 [604-632] AEVRQHREARPALLTSRLRFIPKPDGLRP 29 a.m. 58. pep-RIA-57 [604-633] AEVRQHREARPALLTSRLRFIPKPDGLRPI 30 years 59. pep-RIA-58 [605-633] EVRQHREARPALLTSRLRFIPKPDGLRPI 29 a.m. 60. pep-RIA-59 [606-633] VRQHREARPALLTSRLRFIPKPDGLRPI 28 years 61. pep-RIA-60 [607-633] RQHREARPALLTSRLRFIPKPDGLRPI 27 a.m. 62. pep-RIA-61 [608-633] QHREARPALLTSRLRFIPKPDGLRPI 26 years 63. pep-RIA-62 [609-633] HREARPALLTSRLRFIPKPDGLRPI 25 years 64. pep-RIA-63 [610-633] REARPALLTSRLRFIPKPDGLRPI 24 hours 65. pep-RIA-64 [611-627] EARPALLTSRLRFIPKP 17 a.m. 66. pep-RIA-65 [611-628] EARPLLTSRRFIPKPD 18 months 67. pep-RIA-66 [611-629] EARPALLTSRLRFIPKPDG 19 a.m. 68. pep-RIA-68 [611-631] EARPALLTSRLRFIPKPDGLR 21 a.m. 69. pep-RIA-69 [611-632] EARPALLTSRLRFIPKPDGLRP 22 years 70. pep-RIA-70 [611-633] EARPALLTSRLRFIPKPDGLRPI 23 a.m. 71. pep-RIA-71 [611-634] EARPALLTSRLRFIPKPDGLRPIV 24 hours 72. pep-RIA-72 [611-635] EARPALLTSRLRFIPKPDGLRPIVN 25 years 73. pep-RIA-73 [611-636] EARPALLTSRLRFIPKPDGLRPIVNM 26 months 74. pep-RIA-74 [611-637] EARPALLTSRLRFIPKPDGLRPIVNMD 27 a.m. 75. pep-RIA-75 [611-638] EARPALLTSRLRFIPKPDGLRPIVNMDY 28 months 76. pep-RIA-76 [611-639] EARPALLTSRLRFIPKPDGLRPIVNMDYV 29 a.m. 77. pep-RIA-77 [611-640] EARPALLTSRLRFIPKPDGLRPIVNMDYVV 30 years 78. pep-RIA-78 [611-625] EARPLATSRLRFIP 15 years 79. pep-RIA-79 [611-624] EARPLLTSRLRFI 14 months 80. pep-RIA-80 [611-623] EARPLATSRLRF 13 months

[0048] [Table 3]

[0049] 81. pep-RIA-81 [611-622] EARPLASTRL 12 months 82. pep-RIA-82 [611-621] EARPLASTRL 11 a.m. 83. pep-RIA-83 [611-620] EARPALLTSR 10 years 84. pep-RIA-84 [611-619] EARPHONES 9 a.m. 85. pep-RIA-85 [611-618] EARLY 8 months 86. pep-RIA-86 [611-617] EAR PALL 7 months 87. pep-RIA-87 [611-616] EARPLUS 6 months 88. pep-RIA-88 [611-615] EARPA 5 years 89. pep-RIA-89 [611-614] EARP 4 months 90. pep-RIA-90 [611-613] EAR 3 years 91. pep-RIA-91 [612-626] ARPALLTSRLRFIPK 15 years 92. pep-RIA-92 [613-626] RPALLTSRLRFIPK 14 months 93. pep-RIA-93 [614-626] PALLTSRLRFIPK 13 months 94. pep-RIA-94 [615-626] ALLTSRLRFIPK 12 months 95. pep-RIA-95 [616-626] LLTSRLRFIPK 11 a.m. 96. pep-RIA-96 [617-626] LTSRLRFIPK 10 aa 97. pep-RIA-97 [618-626] TSRLRFIPK 9 aa 98. pep-RIA-98 [619-626] SRLRFIPK 8 aa 99. pep-RIA-99 [620-626] RLRFIPK 7 aa 100. pep-RIA-100 [621-626] LRFIPK 6 aa 101. pep-RIA-101 [622-626] RFIPK 5 aa 102. pep-RIA-102 [623-626] FIPK 4 aa 103. pep-RIA-103 [624-626] IPK 3 aa 104. pep-RIA-104 [612-625] ARPALLTSRLRFIP 14 aa 105. pep-RIA-105 [613-624] RPALLTSRLRFI 12 aa 106. pep-RIA-106 [614-623] PALLTSRLRF 10 aa 107. pep-RIA-107 [615-622] ALLTSRLR 8 aa 108. pep-RIA-108 [616-621] LLTSRL 6 aa 109. pep-RIA-109 [617-620] LTSR 4 aa 110. pep-RIA-110 [612-624] ARPALLTSRLRFI 13 aa 111. pep-RIA-111 [612-623] ARPALLTSRLRF 12 aa 112. pep-RIA-112 [612-622] ARPALLTSRLR 11 aa 113. pep-RIA-113 [612-621] ARPALLTSRL 10 aa 114. pep-RIA-114 [612-620] ARPALLTSR 9 aa 115. pep-RIA-115 [612-619] ARPALLTS 8 aa 116. pep-RIA-116 [612-618] ARPALLT 7 aa 117. pep-RIA-117 [612-617] ARPALL 6 aa 118. pep-RIA-118 [612-616] ARPAL 3 aa 119. pep-RIA-119 [612-615] ARPA 4 aa 120. pep-RIA-120 [612-614] ARP 3 aa

[0050] [Table 4]

[0051] 121 pep-RIA-121 [613-625] RPALLTSRLRFIP 13 aa 122 pep-RIA-122 [613-623] RPALLTSRLRF 11 aa 123 pep-RIA-123 [613-622] RPALLTSRLR 10 aa 124 pep-RIA-124 [613-620] RPALLTSR 8 aa 125 pep-RIA-125 [613-619] RPALLTS 7 aa 126 pep-RIA-126 [613-618] RPALLT 6 aa 127 pep-RIA-127 [613-617] RPALL 5 aa 128 pep-RIA-128 [613-616] RPAL 4 aa 129 pep-RIA-129 [613-615] RPA 3 aa 130 pep-RIA-130 [614-625] PALLTSRLRFIP 12 aa 131 pep-RIA-131 [614-624] PALLTSRLRFI 11 aa 132 pep-RIA-132 [614-622] PALLTSRLR 9 aa 133 pep-RIA-133 [614-621] PALLTSRL 8 aa 134 pep-RIA-134 [614-620] PALLTSR 7 aa 135 pep-RIA-135 [614-619] PALLTS 6 aa 136 pep-RIA-136 [614-618] PALLT 5 aa 137 pep-RIA-137 [614-617] PALL 4 aa 138 pep-RIA-138 [614-616] PAL 3 aa 139 pep-RIA-139 [615-625] ALLTSRLRFIP 11 aa 140 pep-RIA-140 [615-623] ALLTSRLRF 9 aa 141 pep-RIA-141 [615-621] ALLTSRL 7 aa 142 pep-RIA-142 [615-620] ALLTSR 6 aa 143 pep-RIA-143 [615-619] ALLTS 5 aa 144 pep-RIA-144 [615-618] ALLT 4 aa 145 pep-RIA-145 [615-617] ALL 3 aa 146 pep-RIA-146 [616-625] LLTSRLRFIP 10 aa 147 pep-RIA-147 [616-624] LLTSRLRFI 9 aa 148 pep-RIA-149 [616-622] LLTSRLR 7 aa 149 pep-RIA-150 [616-620] LLTSR 5 aa 150 pep-RIA-151 [616-619] LLTS 4 aa 151 pep-RIA-152 [616-618] LLT 3 aa 152 pep-RIA-153 [617-625] LTSRLRFIP 9 aa 153 pep-RIA-154 [617-624] LTSRLRFI 8 aa 154 pep-RIA-155 [617-623] LTSRLRF 7 aa 155 pep-RIA-156 [617-622] LTSRLR 6 aa 156 pep-RIA-157 [617-621] LTSRL 5 aa 157 pep-RIA-158 [617-619] LTS 3 aa 158 pep-RIA-159 [618-625] TSRLRFIP 8 aa 159 pep-RIA-160 [618-624] TSRLRFI 7 aa 160 pep-RIA-161 [618-623] TSRLRF 6 aa

[0052] [Table 5]

[0053]

[0054] In one embodiment of the invention, a polynucleotide encodes a peptide comprising at least one amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, a peptide having at least 80% homology to the aforementioned sequence, or a peptide as a fragment of the aforementioned peptide. The polynucleotide described above enables the production of the peptide in large quantities. For example, culturing a vector comprising a polynucleotide encoding the peptide results in the production of the peptide in large quantities.

[0055] The peptides disclosed herein may include peptides comprising amino acid sequences having homology of more than 80%, more than 85%, more than 90%, more than 95%, more than 96%, more than 97%, more than 98%, or more than 99%. Furthermore, the peptides disclosed in this invention may include: peptides or fragments of peptides comprising any of the amino acid sequences of SEQ ID NO:2 to SEQ ID NO:178, and peptides having more than 1 transformed amino acid, more than 2 transformed amino acids, more than 3 transformed amino acids, more than 4 transformed amino acids, more than 5 transformed amino acids, more than 6 transformed amino acids, or more than 7 transformed amino acids.

[0056] In one embodiment of the invention, changes in the amino acid sequence constitute a change in the physical and chemical properties of the peptide. For example, amino acid conversion can be performed to improve the peptide's thermal stability, alter substrate specificity, and change its optimal pH.

[0057] The term "amino acid" herein includes not only the 22 standard amino acids naturally introduced into peptides, but also D-isomers and transformed amino acids. Therefore, in certain embodiments of the invention, peptides herein include peptides having D-amino acids. On the other hand, peptides may include non-standard amino acids, such as those that have undergone post-translational modifications. Examples of post-translational modifications include phosphorylation, glycosylation, acylation (including acetylation, myristylation, palmitoylation), alkylation, carboxylation, hydroxylation, glycosylation, biotinylation, ubiquitination, transformation of chemical properties (e.g., β-removal of deimide, deamidation), and structural transformation (e.g., formation of disulfide bonds). Similarly, modifications of amino acids are included, which are altered due to chemical reactions during combination with cross-linking agents to form peptide conjugates.

[0058] The peptides disclosed herein may be wild-type peptides that have been identified and isolated from natural sources. Alternatively, when compared to peptide fragments containing any of the amino acid sequences in SEQ ID NO:2 to SEQ ID NO:178, the peptides disclosed herein may be artificial mutant strains comprising one or more amino acids that have been substituted, deleted, and / or inserted. Amino acid variations in wild-type peptides (not only in artificial mutants) include conserved substitutions of amino acids that do not significantly affect the folding and / or activity of the active protein. Examples of conserved substitutions belong to the group consisting of basic amino acids (arginine, lysine, and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine, valine, and methionine), aromatic amino acids (phenylalanine, tryptophan, and tyrosine), and small amino acids (glycine, alanine, serine, and threonine). Amino acid substitutions that do not typically alter specific activity are known in the art. The most common substitutions are alanine / serine, valine / isoleucine, aspartic acid / glutamic acid, threonine / serine, alanine / glycine, alanine / threonine, serine / asparagine, alanine / valine, serine / glycine, tyrosine / phenylalanine, alanine / proline, lysine / arginine, aspartic acid / asparagine, leucine / isoleucine, leucine / valine, alanine / glutamic acid, aspartic acid / glycine, and the reverse. Another example of conserved substitutions is shown in Table 6 below.

[0059] [Table 6]

[0060]

[0061] The essential transformation of the biological properties of peptides is performed by selecting substitutions with significantly different functions: (a) maintaining the polypeptide backbone structure in the substitution region, such as sheet-like or three-dimensional helical structures; (b) maintaining the charge or hydrophobicity of the molecule in the target region; or (c) maintaining the integrity of the side chain. Natural residues are grouped according to general side chain properties as follows:

[0062] (1) Hydrophobic: Leucine, methionine, alanine, valine, leucine, isoleucine;

[0063] (2) Neutral hydrophilic: cysteine, serine, threonine;

[0064] (3) Acids: Aspartic acid, glutamic acid;

[0065] (4) Alkaline: Asparagine, glutamine, histidine, lysine, arginine;

[0066] (5) Residues affecting chain orientation: glycine, proline; and

[0067] (6) Aromatic: tryptophan, tyrosine, phenylalanine.

[0068] Non-conservative substitutions can be performed by replacing the aforementioned types of members with different types. Any cysteine ​​residues that are not related to maintaining the proper three-dimensional structure of the peptide can usually be substituted with serine, thus increasing the oxidative stability of the molecule and preventing inappropriate cross-linking. Conversely, improvements in stability can be achieved by adding one or more cysteine ​​bonds to the peptide.

[0069] The modified type of an amino acid variant of a peptide refers to those amino acids whose antibody glycosylation has been altered. The term "modification" in this document refers to the deletion of at least one sugar residue found in the peptide and / or the addition of at least one glycosylated residue not present in the peptide.

[0070] Glycosylation in peptides is typically N-linked or O-linked. The term "N-linked" in this context refers to attaching a sugar residue to the side chain of an asparagine residue. As tripeptide sequences, asparagine-X-serine and asparagine-X-threonine (where X is any amino acid other than proline) are recognition sequences for enzymatically attaching sugar residues to the side chain of asparagine. Therefore, when one of these tripeptide sequences is present in the polypeptide, a potential glycosylation site is created. "O-linked glycosylation" means attaching one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyl amino acid. The hydroxyl amino acid is usually serine or threonine, but 5-hydroxyproline or 5-hydroxylysine can also be used.

[0071] Adding a glycosylation site to a peptide can be conveniently performed by altering the amino acid sequence to include the aforementioned tripeptide sequence (the glycosylation site for N-linkage). This alteration can be made by adding at least one serine or threonine residue to the first antibody sequence or by substituting those residues (the glycosylation site for O-linkage).

[0072] In one embodiment of the invention, a cell-penetrating peptide comprising a peptide is provided, wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, the peptide having an amino acid sequence having more than 80% homology to the above sequences, or the peptide is a fragment of the above peptide.

[0073] In one embodiment of the invention, a pharmaceutical composition is provided comprising a peptide as a drug delivery system for delivering one or more active ingredients, wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, the peptide having more than 80% homology to the above sequences, or the peptide is a fragment of the above peptide.

[0074] Peptides containing any of the amino acid sequences in SEQ ID NO:2 to SEQ ID NO:178, fragments of the above peptides, or peptides having more than 80% homology with the above sequences are safe and have significant efficacy as cell-penetrating peptides. Therefore, peptides can be conjugated to drugs to deliver drugs inside cells.

[0075] In one embodiment of the invention, a conjugate of a peptide with an active ingredient to be delivered is provided, wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, and the peptide is a fragment of the aforementioned peptide or has more than 80% homology with the aforementioned peptide. In one embodiment of the invention, the active ingredient may be at least one selected from the group consisting of proteins, nucleic acids, peptides, lipids, glycolipids, minerals, sugars, contrast materials, pharmaceuticals, and compounds. In one embodiment of the invention, the active ingredient may be a peptide. In one embodiment of the invention, the active ingredient may be a cytokine, antibody, antibody fragment, therapeutic enzyme, soluble receptor, or ligand.

[0076] The term "cell-penetrating peptide" as used herein refers to peptides capable of delivering cargo into cells both in vitro and / or in vivo. The term "cargo" as used herein encompasses all substances that can be delivered into cells through conjugation with cell-penetrating peptides, such as any substance intended to enhance cell-penetrating efficacy. Specifically, this includes active ingredients in pharmaceuticals, cosmetics, or health foods. More specifically, it includes substances that cannot be delivered into cells via conventional routes, such as sugars, nanoparticles, biologics, viruses, contrast agents, or other compounds, which may include, for example, proteins, nucleic acids, peptides, minerals, glucose, but are not limited to those substances. The term "pharmaceutical" as used herein is a broad concept, encompassing substances intended to be delivered for the purpose of alleviating, preventing, treating, or diagnosing diseases, injuries, or specific symptoms.

[0077] The “carrier peptide” revealed in this article is a peptide that can deliver active ingredients to target sites through conjugation with active ingredients.

[0078] In one embodiment of the invention, the protein or peptide as cargo comprises one or more of the following: hormones, hormone analogs, enzymes, enzyme inhibitors, signal transfer proteins (or peptides), antibodies, and vaccines, but is not limited to those substances. In one embodiment of the invention, the nucleic acid is a molecule that can be spontaneous or artificial, single-stranded or double-stranded DNA or RNA. The nucleic acid molecule can be one or more nucleic acids of the same type (e.g., having the same nucleotide sequence) or different types of nucleic acids. The nucleic acid molecule comprises one or more of the following: DNA, complementary DNA (cDNA), decoy DNA, RNA, small interfering RNA (siRNA), microRNA (miRNA), small hairpin RNA (shRNA), hourly sequence RNA (stRNA), small nucleolar RNA (snoRNA), small nuclear RNA (snRNA), pentose nucleic acid (PNA), antisense oligomers, plasmids, and other modified nucleic acids, but is not limited to those substances. In one embodiment of the invention, the virus comprises the whole virus or a viral core comprising the nucleic acid of the virus. In one embodiment of the invention, the chemical substance is a broad indicator comprising a natural or synthetic substance that can act as a drug.

[0079] The phenomenon where specific DNA expression is controlled by double-stranded RNA (dsRNA) during DNA expression is called RNA interference (RNAi). This phenomenon was first discovered in *C. nematode* in 1998 and has been found to be common in plants, fruit flies, and mammals (Fire et al., *Nature*, 391:806-811, 1998; Novina & Sharp, *Nature*, 430:161-164, 2004).

[0080] RNA interference is regulated by 19-25 bp dsRNAs that enter the cell and then bind to the RNA-induced silencing complex (RISC). The binding of the dsRNA to the antisense strand of the complementary messenger RNA (mRNA) sequence triggers the degradation of the target messenger RNA via endonucleases found within the RISC complex (Rana, TM, Nat. Rev. Mol. Cell Biol., 8:23-36, 2007; Tomari, Y. and Zamore, PD, Genes Dev., 19:517-529, 2005). In other words, small interfering RNAs are included in RNA interference by inhibiting the production of specific proteins and thus interfering with DNA expression. Small interfering RNAs, consisting of 19-23 nucleotides, form base pairs according to the complementary sequence of the messenger RNA to the specific mRNA that forms the double-stranded RNA. Subsequently, the double-stranded RNA is specifically degraded as the messenger RNA is removed from the cell. Small interfering RNA (sRNA) has attracted attention as a substance for gene therapy because it has shown significant effects in inhibiting the expression of specific DNA molecules in recent animal studies. Over the past 20 years, sRNAs with high DNA activation and precise selectivity have been studied, with the aim of replacing antisense oligonucleotides currently used as therapeutics. Therefore, many pharmaceutical companies are now developing sRNA-based therapeutics. Compared to existing antisense oligonucleotides, sRNAs are known to inhibit gene expression by up to 10 times less and to inhibit target genes only with significant gene selectivity. sRNA technology, particularly for therapeutic purposes, offers significant benefits because it is easier to design than other drugs and possesses properties such as high target selectivity and inhibition of specific gene expression. Similarly, because gene expression inhibition via RNA interference utilizes naturally occurring mechanisms in vivo, toxicity is very low. However, sRNAs have the disadvantage of not being easily delivered into cells because they are anionic and cannot penetrate cell membranes, and are easily degraded within a short timeframe due to their low in vivo stability. This disadvantage of sRNAs can be overcome by conjugation with the carrier peptides disclosed in this paper.

[0081] In one embodiment of the invention, the active ingredient (goods) is effective as cancer cells, immune cells, or fibroblasts. Specifically, the cancer cells include any one of the following cell groups: liver cancer cells, breast cancer cells, and leukemia cells; the immune cells include any one of the following cell groups: T lymphocytes, B cells, and monocytes.

[0082] In one embodiment of the present invention, the active ingredient is located in the cytoplasm, and the carrier peptide locally delivers the active ingredient to the cytoplasm.

[0083] In one embodiment of the present invention, the active ingredient is located in the mitochondria, and the carrier peptide locally delivers the active ingredient to the mitochondria.

[0084] In one embodiment of the invention, a drug delivered inside a cell via a cell-penetrating peptide may include one or more drug delivery carriers, such as lipid particles, micelles, nanoparticles, magnetic particles, or quantum dots.

[0085] The term "contrast material" as used herein is a broad designation encompassing all substances used to contrast structures or fluids within the body in medical imaging. Appropriate contrast materials include, but are not limited to, radiopaque contrast agents, paramagnetic contrast agents, superparamagnetic contrast agents, computed tomography (CT) contrast materials, and other contrast materials. For example, radiopaque contrast agents (used in X-ray imaging) may include inorganic and organic iodine compounds (e.g., diatrizoate), radiopaque metals and salts of such metals (e.g., silver, gold, platinum, etc.), and other radiopaque compounds (e.g., calcium salts, barium salts such as barium sulfate, tantalum, and tantalum oxide). Suitable paramagnetic contrast materials (for MR imaging) include gadolinium diethylene triaminepentaacetic acid (Gd-DTPA) and its derivatives, as well as other gadolinium, manganese, iron, dysprosium, copper, europium, erbium, chromium, nickel, and cobalt complexes, such as 1,4,7,10-tetraazacyclododecan-N,N',N”,N”'-tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA), and 1,4,7,10-tetraazacyclododecan-N,-N',N”'-triacetic acid (TTA). (DO3A), 1,4,7-triazacyclononane-N,N',N”-TRIACETIC ACID (NOTA), 1,4,8,10-tetraazacyclotetradecane-N,N',N”,N”'-tetraacetic acid (TETA), and hydroxybenzylethylene-diamine diacetic acid (HBED). Suitable superparamagnetic contrast materials (for MR imaging) include magnetite, superparamagnetic iron oxide (SPIO), ultrasmall superparamagnetic iron oxide (USPIO), and single-crystal iron oxide.Other suitable contrast agents are iodinated, non-iodinated, ionic and non-ionic CT contrast agents, contrast agents similar to rotational markers or diagnostically effective agents.

[0086] Other examples of contrast agents include β-galactosidase, green fluorescent protein, cyan fluorescent protein, luciferase (but not limited to those substances), and marker genes encoding proteins that are readily detectable when expressed in cells. Various markers can be used, such as radioactive isotopes, flours, enzymes, enzyme substrates, enzyme cofactors, enzyme inhibitors, and ligands (especially heptans).

[0087] In one embodiment of the present invention, the contrast agent is ferrocene carboxylic acid, hereinafter referred to as Formula 2. The structure of ferrocene is represented in Formula 1.

[0088] [Chemical Formula 1]

[0089]

[0090] [Chemical Formula 2]

[0091]

[0092] In one embodiment of the invention, the conjugate of the cell-penetrating peptide and the contrast contrast material is ferrocene carboxyl-pep1, represented by chemical formula 3 below.

[0093] [Chemical Formula 3]

[0094]

[0095] In one embodiment of the invention, the peptide or composition may be fused with one or more detectable tags. The tags may be compounds detectable in chemical, physical, or enzymatic reactions, or compounds that generate a signal directly or indirectly in a reaction. Tagging and detection may then be performed according to methods well known in the art (e.g., Sambrook, J. and Russell, DW (2001); and Lottspeich, F. and Zorbas H. (1998) Bioanalytik, Spektrum Akademischer Verlag, Heidelberg / Berlin, Germany). Tags include, but are not limited to, fluorescent tags, enzyme tags, chromogenic tags, luminescent tags, radiation tags, haptens, biotin, metal complexes, metals, and colloidal gold. All forms of such tags are well known in the art and are available from various suppliers.

[0096] In one embodiment of the invention, the cargo can be directly bound to the peptide. In another embodiment of the invention, the cargo can be bound to the peptide via various types of bonds, such as covalent or non-covalent bonds. For example, in one embodiment of the invention, the cargo can be bound to the N-terminus or C-terminus of the peptide. For instance, the cargo can be bound to the peptide via disulfide bonds or covalent bonds. A covalent bond is a bond that can bind the cargo to an α-amine of the N-terminal glutamic acid residue or an amine of the C-terminal lysine residue. Similarly, the peptide and the cargo can be bound via non-covalent bonds, which allows the peptide or the cargo to be encapsulated in a capsule form.

[0097] In another embodiment of the invention, the peptide can bind to the cargo via a linker. For example, the peptide can bind to the cargo by introducing a linker, such as a hydrazinopyridine-3-carboxylic acid linker, into an α-amine of an N-terminal glutamic acid residue or an amine of a C-terminal lysine residue.

[0098] In another embodiment of the invention, when the cargo is DNA or RNA, an SH group (thiol group) is introduced into the peptide and a maleimide group is introduced into the DNA or RNA. Subsequently, the SH group of the peptide and the maleimide group of the DNA or RNA bind, thus creating a binding between the cargo and the peptide.

[0099] In another embodiment of the invention, when the cargo is a peptide or protein, the DNA expressing the cargo binds to the DNA expressing the vector peptide, and by expressing this DNA binding, the cargo and peptide can be combined into a fusion protein. A specific example of fusion protein binding is as follows: when primers are prepared to generate the fusion protein, the nucleotide encoding the vector peptide is attached to the front of the nucleotide expressing the cargo, and the resulting nucleotide is inserted into a vector such as a polyethylene terephthalate (pET) vector using a restriction enzyme, and the nucleotide is expressed by transformation into cells such as BL-21(DE3). In this case, the fusion protein will be effectively expressed by treating the fusion protein with an expression-inducing agent similar to isopropyl-1-thio-β-D-galactopyranoside (IPTG). Subsequently, the expressed fusion protein line was purified by His-tagged purification, dialyzed with PBS, and then concentrated by centrifugation at 2000 rpm to 4000 rpm for 5 to 20 minutes by adding it to a kit.

[0100] In one embodiment of the invention, the carrier peptide binds to a staining substance, a fluorescent substance, particularly fluorescein isothiocyanate (FITC) or green fluorescent protein (GFP). In one embodiment of the invention, FITC binds to the amino group (NH3+) of a lysine residue at the N-terminus or C-terminus of the carrier peptide. In the case of peptides in which lysine residues are not present at the peptide terminus, the peptide can bind to FITC via a linker including a lysine residue.

[0101] The carrier peptide disclosed herein can bind to the cargo in a 1:1 molar ratio, but it can also bind to the cargo in a different molar ratio. The carrier peptide is a peptide containing any of the amino acid sequences in SEQ ID NO:2 to SEQ ID NO:178, or a peptide having an amino acid sequence with more than 80% homology to the aforementioned peptides, or a fragment of the aforementioned peptides. For example, the molar ratio of CPP to cargo can be greater than 2:1, specifically greater than 2:1, greater than 3:1, greater than 4:1, greater than 5:1, greater than 6:1, greater than 7:1, greater than 8:1, greater than 9:1, or greater than 10:1. This means that a large number of carrier peptide molecules can bind to cargo molecules. A large number of carrier peptide molecules can bind sequentially or in parallel. "Sequential binding" means that the carrier peptide and cargo molecule will bind at the terminal amino acid. "Parallel binding" means that the carrier peptide and cargo molecule will bind at a site different from the terminal amino acid. On the other hand, the molar ratio of carrier peptide to cargo can be greater than 1:2. This means that the carrier peptide molecule can bind to a large number of cargo molecules. For example, the molar fraction of the carrier peptide to the cargo can be 1:2, specifically, greater than 1:2, greater than 1:3, greater than 1:4, greater than 1:5, greater than 1:6, greater than 1:7, greater than 1:8, greater than 1:9 or greater than 1:10.

[0102] The movement pathways of peptides bound to fluorescein isothiocyanate can be easily identified. Therefore, the carrier peptide in one embodiment of the present invention will be used for cell imaging or to detect drug delivery pathways within cells.

[0103] In one embodiment of the invention, the use of a peptide as a drug delivery carrier for delivering more than one active ingredient is provided, wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, or the peptide is a fragment of the aforementioned peptide, or the peptide has an amino acid sequence having more than 80% homology with the aforementioned peptide. The use may refer to therapeutic or non-therapeutic purposes.

[0104] In one embodiment of the invention, a method for delivering a drug into the cells of a test subject is provided, the method comprising the step of administering a composition comprising the drug and a peptide; wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, or the peptide is a fragment of the aforementioned peptide, or the peptide has an amino acid sequence having more than 80% homology with the aforementioned peptide.

[0105] In one embodiment of the present invention, a method for detecting a drug delivery pathway is provided, the method comprising the steps of applying a peptide and a contrast material to a test subject; wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, or the peptide is a fragment of the aforementioned peptide, or the peptide has an amino acid sequence having more than 80% homology with the aforementioned peptide.

[0106] In one embodiment of the present invention, a method for detecting a drug delivery pathway is provided, the method comprising applying a conjugate of a peptide and a contrast material to a test subject; wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, or the peptide is a fragment of the aforementioned peptide, or the peptide has an amino acid sequence having more than 80% homology with the aforementioned peptide.

[0107] In one embodiment of the invention, a kit is provided for delivering a drug into the cells of a test subject. The kit contains a composition and instructions for use, wherein the composition comprises a conjugate of the peptide of the invention with the drug for delivery, wherein the peptide comprises any amino acid sequence of SEQ ID NO:2 to SEQ ID NO:178 or the peptide is a fragment of the aforementioned peptide, or the peptide has an amino acid sequence having more than 80% homology with the aforementioned peptide, and wherein the instructions for use include at least one of the following: dosage, route of administration, frequency of administration, and instructions for the composition.

[0108] In one embodiment of the present invention, a cosmetic or food composition comprising an active ingredient and a peptide is provided; wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, the peptide has an amino acid sequence having more than 80% homology to the aforementioned sequences, or the peptide is a fragment of the aforementioned peptide. In another embodiment of the present invention, a cosmetic or food composition comprising a conjugate of a peptide and an active ingredient is provided; wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, the peptide has an amino acid sequence having more than 80% homology to the aforementioned sequences, or the peptide is a fragment of the aforementioned peptide.

[0109] In one embodiment of the invention, a pharmaceutical, cosmetic, or food composition is provided having a significant ability to deliver an active ingredient into cells, the pharmaceutical, cosmetic, or food composition comprising a conjugate of a peptide and an active ingredient; wherein the peptide comprises any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, the peptide having an amino acid sequence having more than 80% homology to the above sequences, or the peptide is a fragment of the above peptide.

[0110] Mitochondria, as the central organelle in energy metabolism in eukaryotic cells, are the first intracellular organelles known to be associated with human diseases (Luft R, Ikkos D, Palmieri G, Ernster L, Afzelius B: A case of severe hypermetabolism of nonthyroid origin with a defect in the maintenance of mitochondrial respiratory control: a correlated clinical, biochemical, and morphological study), J Clin Invest 41: 1776-804, 1962).

[0111] Because mitochondria play a crucial role in controlling cellular energy metabolism and apoptosis, they serve as primary targets for various therapeutic drugs. Similarly, this organelle is involved in controlling intracellular calcium concentration, the mitochondrial respiratory chain acts as a vital electron transport system in energy production, and it leads to the production of reactive oxygen species. Therefore, abnormal mitochondrial function is closely related to adult diseases such as diabetes insipidus, cardiomyopathy, infertility, blindness, kidney / liver disease, and stroke (Modica-Napolitano KS, Singh KK: Mitochondria as targets for detection and treatment of cancer. Expert Rev Mol Med 11:1-19, 2002). Likewise, it is being proposed to include mitochondrial genetic mutations in the development of aging, degenerative neuronal diseases, and cancer.

[0112] The mitochondrial targeted delivery system provided according to one embodiment of the present invention may include any of the above-described conjugates, wherein the carrier peptide locally moves into the mitochondria and performs the function of locally delivering the active ingredient into the intracellular mitochondria, wherein the peptide and fragment having an amino acid sequence having at least 80% homology with the above-described sequence are peptides that maintain the mitochondrial targeted delivery system, and the mitochondrial targeted peptide may be a peptide having any of the amino acid sequences in SEQ ID NO:2 to SEQ ID NO:178.

[0113] A mitochondrial activity modulating composition may be provided, wherein the composition comprises a conjugate of the peptide of the present invention and a delivery carrier peptide, wherein the carrier peptide locally moves into the intracellular mitochondria and performs the function of locally delivering the active ingredient into the intracellular mitochondria, wherein the peptide and fragment of the peptide having an amino acid sequence having at least 80% homology with the above sequence are peptides that maintain a mitochondrial targeted delivery system, and the mitochondrial targeted peptide may be a composition having any of the amino acid sequences of SEQ ID NO:2 to SEQ ID NO:178.

[0114] One embodiment of the present invention provides a mitochondrial activity regulating composition, which is used as a pharmaceutical composition to treat mitochondrial-related diseases or conditions, prevent or inhibit disease progression, or relieve symptoms; wherein the active ingredient is used to treat mitochondrial-related diseases or conditions, prevent or inhibit disease progression, or relieve symptoms.

[0115] This article reveals that "mitochondrial-related diseases" include Huntington's disease, amyotrophic lateral sclerosis (ALS), mitochondrial encephalomyopathy with lactic acidemia and stroke-like episodes (MELAS); myoclonus, epilepsy, and myopathy with ragged red fibers (MERRF); neuromuscular weakness, ataxia, retinitis pigmentosa / Maternally inherited leigh syndrome (NARP / MILS); Leber's hereditary optic neuropathy (LHON); Kearns-Sayre syndrome (KSS); and Pearson's myelopancreatitis. Marrow-Pancreas Syndrome (PMPS); Chronic progressive external ophthalmoplegia (CPEO); Reye's syndrome; Alpert syndrome; Multiple mitochondrial DNA deletion syndrome; Mitochondrial DNA depletion syndrome; Complex I deficiency; Complex II (succinate dehydrogenase (SDH)) deficiency; Complex III deficiency; Cytochrome c oxidase (COX, Complex IV) deficiency; Complex V deficiency; Adenine nucleotide transporter (Adenine nucleotide) Translocator (ANT) deficiency; pyruvate dehydrogenase (PDH) deficiency; ethylmalonic aciduria with lactic acidemia; 3-methylpentenic aciduria with lactic acidemia; irritable epilepsy manifested as attenuation during infection; Asperger's syndrome manifested as attenuation during infection; autism manifested as attenuation during infection; attention deficit hyperactivity disorder (ADHD); cerebral palsy manifested as attenuation during infection; alexia manifested as attenuation during infection; maternally inherited thrombocytopenia; leukemia; MNGIE (mitochondrial myopathy, peripheral and autonomic neuropathy, gastrointestinal dysfunction, and epilepsy);MARIAHS syndrome (mitochondrial dysregulation, recurrent infections, aphasia, hypouricemia / myelin deficiency, seizures, and dicarboxylic aciduria); ND6 dystonia; cyclic vomiting manifested as a decrease in intensity during infection; 3-hydroxyisobutyric aciduria with lactic acidosis; diabetes insipidus with lactic acidosis; uridine reactive neural syndrome (URNS); familial bilateral striatum necrosis (FBSN); hearing loss associated with aminoglycosides; relaxed cardiomyopathy; splenic lymphoma; tungsten symptoms; symptoms of multiple mitochondrial DNA deletions; and symptoms of renal tubular acidosis / diabetes insipidus / dysregulation, but not limited to those diseases.

[0116] In another embodiment of the invention, a nucleic acid molecule encoding the aforementioned polypeptide is provided. For example, the nucleic acid molecule has the base sequence GAA GCG CGC CCG GCG CTG CTG ACC AGC CGC CTG CGC TTT ATT CCG AAA. The nucleic acid can be introduced into a host cell using methods well known to those skilled in the art. For example, well-known methods include transformation methods such as calcium phosphate methods, liposomes, electroporation, contact with viruses and cells, or direct microinjection into cells. The host cell can be a higher eukaryotic cell (e.g., mammalian cell), a lower eukaryotic cell (such as yeast cell), or a prokaryotic cell (such as bacterial cell). Suitable prokaryotic host cells for transformation can be species belonging to, for example, *Escherichia coli*, *Bacillus subtilis*, *Salmonella*, *Pseudomonas*, *Streptomyces*, and microbacterial species.

[0117] The vectors containing the aforementioned nucleic acid molecules are typically recombinant expression vectors, and these vectors contain a replication origin that enables host cell transformation and optional markers (e.g., dihydrofolate reductase for eukaryotic cell culture, or tolerance of neomycin, tetracycline or ampicillin in Escherichia coli, yeast TRP1 gene), and a promoter for transcription that controls the protein spreading sequence. For example, the expression vectors that can be used include: well-known bacterial plasmids, such as SV40, derivatives of pcDNA; and well-known bacterial plasmids, such as colE1, pCR1, pBR322, pMal-C2, pET, pGEX (Smith et al., Gene 67:31-40 (1988)); plasmids, such as pMB9 and pMB9 derivative RP4; phage DNA identical to numerous derivatives of phage I, such as NM989; phage DNA such as M13 and filamentous single-stranded phage DNA; yeast plasmids, for example, phage DNA or vectors induced by a combination of modified plasmids with expression repressor sequences. Mammalian expression vectors contain an origin of replication, a suitable promoter, and an enhancer. Similarly, the vector may contain a forced ribosome binding site, a polyadenylation site, splicing donor and acceptor portions, a transcription termination sequence, and a 5' planking non-transcriptional sequence. Mammalian expression vectors may contain inducible promoters, such as vectors containing dihydrofolate reductase promoters, or any expression vector containing a DHFR expression cassette or a DHFR / methotrexate co-amplification vector, such as pED (Randal J. Kaufman, 1991, Randal J. Kaufman, Current Protocols in Molycular Biology, 16, 12 (1991)). Alternatively, vectors may be used that are glutamine synthetase / methionine sulfonamide co-amplification vectors, such as pEE14 (Celltech), human herpesvirus type 4 (Epstein-Barr-Virus; EBV); or vectors that guide epitype expression under nuclear antigen (EBNA) control, such as pREP4 (Invitrogen), pCEP4 (Invitrogen), pMEP4 (Invitrogen), pREP8 (Invitrogen), pREP9 (Invitrogen), and pEBVHis (Invitrogen). Mammalian expression vectors such as Rc / CMV (Invitrogen) and pRc / RSV (Invitrogen) can be selected. Vaccinia virus mammalian expression vectors that can be used in this invention include pSC11, pMJ601, and pTKgptF1S.

[0118] The yeast expression vector system used in this invention includes non-fusion pYES2 vector (Invitrogen), fusion pYESHisA, B, C (Invitrogen), pRS vector, etc.

[0119] The aforementioned vectors can be introduced into various cell types, such as mammalian cells (especially human-derived cells) or bacterial, yeast, fungal, insect, nematode, and plant cells. Examples of suitable cell types include: VERO cells; HELA cells, such as ATCC No. CCL2; CHO cell lines, such as ATCC No. CCL61; COS cells, such as COS-7 cells and ATCC No. CRL 1650 cells; W138, BHK, HepG2, and 3T3, such as ATCC No. CRL6361; A549, PC12, and K562 cells; 293 cells; Sf9 cells, such as ATCC No. CRL1711; and Cv1 cells, such as ATCC No. CCL70, etc.

[0120] Other suitable cells used in this invention are prokaryotic host cell strains, such as strains belonging to Escherichia coli (e.g., DH5-α strain), Bacillus subtilis, Salmonella, Pseudomonas, Streptomyces and Staphylococcus.

[0121] In one embodiment of the invention, the composition may contain 0.1 μg / mg to 1 mg / mg, specifically 1 μg / mg to 0.5 mg / mg, more specifically 10 μg / mg to 0.1 mg / mg of the following peptides: peptides comprising any amino acid sequence from SEQ ID NO:2 to SEQ ID NO:178, peptides comprising amino acid sequences having more than 80% homology to the above sequences, or fragments of the above peptides. When the contained peptides are within the above ranges, all safety and stability requirements of the composition are met and it is suitable in terms of cost-effectiveness.

[0122] In one embodiment of the invention, the composition can be applied to all animals, including humans, dogs, chickens, pigs, cows, sheep, guinea pigs, and monkeys.

[0123] In one embodiment of the invention, the pharmaceutical composition may be administered via bone marrow, epidural or subcutaneous means, orally, rectally, transdermally, intravenously, intramuscularly or intraperitoneally.

[0124] Oral administration may take the form of, but is not limited to, tablets, pills, soft capsules or hard capsules, granules, powders, solutions or emulsions. Non-oral administration may take the form of, but is not limited to, injections, drops, lotions, ointments, gels, creams, suspensions, aqueous emulsions, suppositories, patches or sprays.

[0125] In one embodiment of the invention, the pharmaceutical composition may, if necessary, contain additives such as diluents, excipients, lubricants, binders, disintegrants, buffers, dispersants, surfactants, colorants, flavorings, or sweeteners. In one embodiment of the invention, the pharmaceutical composition may be manufactured using conventional industrial methods in this art.

[0126] In one embodiment of the invention, the active ingredient of the medical composition may vary depending on factors such as the patient's age, sex, weight, pathology and condition, route of administration, or the prescribing physician's judgment. The dosage based on these factors is determined within the skill of a person skilled in the art, and the daily dose may be, for example, but not limited to, 0.1 μg / kg / day to 1 g / kg / day, specifically 1 μg / kg / day to 10 mg / kg / day, more specifically 10 μg / kg / day to 1 mg / kg / day, and more specifically 50 μg / kg / day to 100 μg / kg / day. In one embodiment of the invention, the pharmaceutical composition may be administered once to three times daily, but is not limited thereto.

[0127] In one embodiment of the invention, the cosmetic composition can be provided in all forms suitable for topical application. For example, this form can be provided as a solution, an aqueous emulsion obtained by dispersion of an oil phase in water, an aqueous emulsion obtained by dispersion of water in an oil phase, a suspension, a solid, a gel, a powder, a paste, a foam, or an aerosol. This form can be manufactured by industrial methods known in the art.

[0128] In one embodiment of the invention, the cosmetic composition may include other ingredients that may desirably enhance the main effect, within a level that does not impair the main effect. In another embodiment, the cosmetic composition may further include emollients, emollients, surfactants, UV absorbers, preservatives, fungicides, antioxidants, pH adjusters, organic or inorganic pigments, fragrances, coolants, or antiperspirants. The formulation ratios of the above ingredients may be determined by those skilled in the art without impairing the purpose and effect of the invention, and the formulation ratios based on the total weight of the cosmetic composition may be from 0.01% to 5% by weight, specifically from 0.01% to 3% by weight.

[0129] In one embodiment of the invention, the food composition is not limited to any particular form, but may be, for example, in granular, powder, liquid, or solid form. Each form may consist of ingredients commonly used in industry (other than the active ingredient), appropriately selected by those skilled in the art, and the effects of having other ingredients may be enhanced.

[0130] The determination of the dosage of the above-mentioned active ingredients is within the level of a person skilled in this technique, and the daily dosage may be, for example, 1 μg / kg / day to 10 mg / kg / day, more specifically 10 μg / kg / day to 1 mg / kg / day, more specifically 50 μg / kg / day to 100 μg / kg / day, but is not limited to this amount and may vary according to age, health status, comorbidities and other various factors.

[0131] The terminology used herein is intended to describe embodiments and not to limit the invention. The numerous terms used above are not intended to limit quantity but rather to indicate that more than one of the terms may exist. The terms “comprising,” “having,” “forming,” and “including” should be interpreted openly (i.e., “including but not limited to”).

[0132] The reference to a range of quantities is used instead of describing separate quantities within a range; therefore, unless explicitly stated otherwise, each quantity can be read as a separate quantity integrated in this document. End values ​​of all ranges are included within the range and can be independently combined.

[0133] Unless otherwise stated or clearly contradicted by the context, all methods mentioned herein may be performed in the appropriate order. Unless included within the scope of the claims, the use of any implementation and all embodiments or exemplary language (e.g., the use of language such as "like...") is intended to more clearly describe the invention and not to limit its scope. Any language used herein outside the scope of the claims shall not be construed as essential to the invention. Unless otherwise defined, the technical and scientific terms used herein have the meaning commonly understood by one skilled in the art to which this invention pertains.

[0134] The preferred embodiments of the present invention are the best mode known to the inventors who practice the invention. This will be clear to those skilled in the art upon reading the statements prior to the variations in the preferred embodiments. The inventors intend that those skilled in the art will fully utilize these variations and carry out the invention in ways other than those listed herein. Therefore, as permitted by patent law, the invention includes equivalents and variations of the key points described in the appended claims. Furthermore, all possible variations within any combination of the foregoing components are included in the invention unless otherwise expressly stated or contradicted by the context. Although the invention has been described and illustrated by exemplary embodiments, those skilled in the art will readily understand that various variations in form and detail defined by the hereinafter claimed claims are possible without departing from the spirit and scope of the invention.

[0135] Example 1: Peptide Synthesis

[0136] The peptides having SEQ ID NO:2 to SEQ ID NO:178 are synthesized according to existing methods of solid-phase peptide synthesis. Specifically, the peptides are synthesized by coupling each amino acid from the C-terminus using ASP48S (Peptron Corporation, Daejeon, South Korea) via Fmoc solid-phase peptide synthesis (SPPS). The peptides are then used such that the first amino acid at the C-terminus of the peptide is attached to the resin as follows:

[0137] NH2-Lysine (Boc)-2-chloro-triphenylmethyl resin

[0138] NH2-alanine-2-chloro-triphenylmethyl resin

[0139] NH2-arginine (Pbf)-2-chloro-triphenylmethyl resin

[0140] All amino acid components of the synthetic peptide are protected at the N-terminus by Fmoc, and the amino acid residues are protected by acid-soluble Trt, Boc, t-Bu (t-butyl ester), and Pbf (2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl). Examples include:

[0141] Fmoc-alanine-OH, Fmoc-arginine (Pbf)-OH, Fmoc-glutamic acid (OtBu)-OH, Fmoc-proline-OH, Fmoc-leucine-OH, Fmoc-isoleucine-OH, Fmoc-phenylalanine-OH, Fmoc-serine (tBu)-OH, Fmoc-threonine (tBu)-OH, Fmoc-lysine (Boc)-OH, Fmoc-glutamine (Trt)-OH, Fmoc-tryptophan (Boc)-OH, Fmoc-methionine-OH, Fmoc-asparagine (Trt)-OH, Fmoc-tyrosine (tBu)-OH, Fmoc-aminocaproic acid-OH, Trt-thioacetic acid.

[0142] HBTU [2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylammonium hexafluorophosphate] / HOBt [N-hydroxybenzotriazine] / NMM [4-methylmorpholine] was used as the coupling agent. Fmoc was removed using dimethylformamide (DMF) containing 20% ​​piperidine. To remove the residue protection or to separate the synthesized peptide from the resin, a splitting mixture [trifluoroacetic acid (TFA) / triisopropylsilane (TIS) / ethanedithiol (EDT) / H2O = 92.5 / 2.5 / 2.5 / 2.5] was used.

[0143] Peptides were synthesized using a solid-phase molecular framework by adding each amino acid in the following sequence: amino acid protection, coupling reaction, washing, and deprotection. After the synthesized peptide was cleaved from the resin, it was purified by high-performance liquid chromatography (HPLC) and the synthesis was verified by mass spectrometry (MS), followed by lyophilization.

[0144] The specific peptide synthesis process is described below using an example of pep1 (EARPALLTSRLRFIPK).

[0145] 1) Coupling

[0146] The amino acid (8 equivalents) protected with NH2-lysine (Boc)-2-chloro-triphenylmethyl resin was melted in the coupling agent HBTU (8 equivalents) / HOBt (8 equivalents) / NMM (16 equivalents), and after the addition of DMF, the reaction mixture was incubated at room temperature for 2 hours, followed by washing with DMF, MeOH and DMF in sequence.

[0147] 2) Fmoc protection

[0148] After adding 20% ​​piperidine to DMF, the reaction mixture was incubated at room temperature for 5 minutes twice, followed by washing with DMF, MeOH and DMF in sequence.

[0149] 3) The basic framework of the peptide is formed by repeatedly performing reactions 1 and 2.

[0150] 4) Cutting: Add the splitting mixture to the fully synthesized peptide and separate the peptide from the resin.

[0151] 5) Add pre-cooled diethyl ether to the mixture, and then centrifuge the reaction mixture to precipitate the peptide.

[0152] 6) After purification by Prep-HPLC, the molecular weight was checked by LC / MS and the peptide was freeze-dried to obtain a powder form.

[0153] Example 2: pep(CPP)-FITC Preparation of conjugates

[0154] (1) Preparation of FITC-CPP conjugates

[0155] The following are examples of conjugates of peptides having SEQ ID NO:2 to SEQ ID NO:178 that are bound to FITC, for example, conjugates of pep1 and FITC, in other words, FITC-connector-pep1 are manufactured as follows.

[0156] The basic framework of the peptide obtained according to the manufacturing method described in Example 1, NH2-linker-E(OtBu)-AR(Pbf)-PALLT(tBu)-S(tBu)-R(Pbf)LR(Pbf)-FIPK(Boc)-2-chloro-triphenylmethyl resin, was reacted with FITC. Specifically, fluorescein-5-isothiocyanate (FITC) (8 equivalents) and N,N-diisopropylethylamine (DIPEA) (16 equivalents) were melted in DMF. A DMF solution was added and the reaction was carried out at room temperature for 2 hours, followed by washing with DMF, MeOH, and DMF in sequence. Thus, FITC-linker-E(OtBu)-AR(Pbf)-PALLT(tBu)-S(tBu)-R(Pbf)LR(Pbf)-FIPK(Boc)-2-chloro-triphenylmethyl resin was obtained. The linker used herein is 6-aminohexanoic acid (Ahx). A TFA / TIS / H2O ratio of 95 / 2.5 / 2.5 was added to the peptide formed on the resin, and the conjugate system was separated from the resin. Pre-cooled diethyl ether was added to the resulting mixture, and the peptide conjugate was precipitated by centrifugation. After purification by Prep-HPLC, the purity was determined by analytical HPLC, and the molecular weight was determined by LC / MS. The peptide system synthesized as described above was verified as FITC-pep1 by molecular weight determination by LC / MS. The conjugate was then freeze-dried. Conjugates of peptides fused to FITC in SEQ ID NO:2 to SEQ ID NO:178 were also prepared in the same manner as described for pep1.

[0157] (2) Preparation of CPP-FITC conjugates

[0158] The basic framework of the peptide (NH2-E(OtBu)-AR(Pbf)-PALLT(tBu)-S(tBu)-R(Pbf)LR(Pbf)-FIPK(Dde)-2-chloro-triphenylmethyl resin) was produced according to the manufacturing method described in Example 21.(1). To selectively introduce FITC to the C-terminus of the peptide, the N-terminus of the peptide was protected from Boc. Subsequently, di-tert-butyl dicarbonate (30 equivalents) and DIPEA (30 equivalents) were melted in DMF. A DMF solution was added to the peptide and incubated at room temperature for 2 hours, and the peptide was washed sequentially with DMF, MeOH, and DMF. Thus, Boc-E(OtBu)-AR(Pbf)-PALLT(tBu)-S(tBu)-R(Pbf)LR(Pbf)-FIPK(Dde)-2-chloro-triphenylmethyl resin was obtained. Dde was removed using DMF containing 2% hydrazine to add FITC to the C-terminus of lysine residues, with the Dde serving as a protecting group for the C-terminal lysine. Subsequently, FITC (8 equivalents) and DIPEA (16 equivalents) were melted in DMF, which was added to the peptide reaction mixture, and the mixture was incubated at room temperature for 2 hours, followed by washing with DMF, MeOH, and DMF in that order. This yielded Boc-E(OtBu)-AR(Pbf)-PALLT(tBu)-S(tBu)-R(Pbf)LR(Pbf)-FIPK(FITC)-2-chloro-triphenylmethyl resin. TFA / TIS / H2O = 95 / 2.5 / 2.5 was added to separate the peptide from the resin. Pre-cooled diethyl ether was added to the mixture, and centrifugation was used to precipitate the peptide. After purification by Prep-HPLC, purity was determined by analytical HPLC, and molecular weight was determined by LC / MS. The obtained substance was confirmed to be pep1-FITC by LC / MS to verify its molecular weight. The conjugate was then freeze-dried. Peptide-FITC conjugates of SEQ ID NO:2 to SEQ ID NO:178 were also prepared in the same manner as described above.

[0159] Example 3: Penetration of ferrocene carboxyl-CPP conjugates

[0160] An amino acid (8 equivalents) protected with NH2-lysine (Boc)-2-chloro-triphenylmethyl resin and coupling agents HBTU (8 equivalents) / HoBt (8 equivalents) / NMM (16 equivalents) were fused in DMF for coupling. DMF solution was added and the reaction was carried out at room temperature for 2 hours, followed by washing with DMF, MeOH, DMF in that order. Subsequently, DMF containing 20% ​​piperidine was added for Fmoc deprotection and the reaction was carried out at room temperature for 5 minutes twice, followed by washing with DMF, MeOH, DMF in that order. By repeating the above reaction, the basic framework of the peptide (NH2-E(OtBu)-AR(Pbf)-PALLT(tBu)-S(tBu)-R(Pbf)LR(Pbf)-FIPK(Dde)-2-chloro-triphenylmethyl resin) was constructed. DMF containing 2% hydrazine was added to remove Dde, which is the protecting group of the C-terminal lysine residue. Subsequently, ferrocene carboxylic acid (Sigma Aldrich cat.#_46264, 16 equivalents) and coupling agents HBTU (16 equivalents) / HoBt (16 equivalents) / NMM (32 equivalents) were melted in DMF. The DMF solution was added and the reaction was carried out at room temperature for 2 hours, followed by washing with DMF, MeOH, and DMF in that order. TFA / TIS / H2O = 95 / 2.5 / 2.5 was added to the synthesized peptide resin to separate the peptide from the resin. Cooling ether was added to the resulting mixture, and the aggregated peptides were precipitated by centrifugation. The precipitate was purified by HPLC and identified by MS. The peptides were then lyophilized.

[0161] Example 4: Cell penetration assay of the pep-FITC conjugate

[0162] (1) Cell penetration experiment in HeLa cell line

[0163] Cell culture

[0164] HeLa cell line, human cervical adenocarcinoma cells, was purchased from ATCC. Cells were cultured at 37°C in a 5% CO2 incubator in minimum essential medium (MEM) containing 10% fetal bovine serum (Invitrogen, USA), Ellis salts, non-essential amino acids, sodium pyruvate, 100 μg / mL penicillin, and 10 units / mL streptomycin.

[0165] Flow cytometry and confocal microscopy analysis that penetrates cells

[0166] Flow cytometry and confocal microscopy were performed to compare the extent of cellular uptake of the peptides, pep (CPP), and control cells as specified in SEQ ID NO:2 to SEQ ID NO:178.

[0167] Cell lines were divided in 6-well plates and cultured at 37°C in a 5% CO2 incubator for 12 hours in medium containing 10% fetal bovine serum (Invitrogen, USA), 100 μg / ml penicillin, and 100 units / ml streptomycin. After washing the cell lines with PBS, they were starved in minimum essential medium for one hour. Each carrier peptide was treated with 20 μM and cultured at 37°C for one hour. After repeating the PBS washing step three times, trypsin-EDTA was treated at 37°C for 10 minutes to separate the carrier peptides outside the cells. Cells were collected with chilled PBS and centrifuged, repeating the cell washing step three times. Subsequently, cells were resuspended in 0.5 ml PBS containing 4% paraformaldehyde, and cell fluorescence was analyzed using FACS Calibur (BD Biosciences, USA). Cell uptake patterns of controls and various peptides bound to FITC were compared and analyzed by mean fluorescence intensity (MFI).

[0168] The result is Figures 2 to 29 It is displayed in the middle. Figure 1 The results of pep 1, with reference to SEQ ID NO:1, are shown. Figures 2 to 29 The analytical results shown are presented in detail in Table 7 below.

[0169] [Table 7]

[0170]

[0171]

[0172]

[0173] (2) Cell penetration in the Huh 7 cell line

[0174] Cell culture

[0175] The Huh7 (human hepatocellular carcinoma) cell line was purchased from the American Type Cell Collection (ATCC) and used as a suspension cell line. The cells were cultured at 37°C in a 5% CO2 incubator in MEM medium containing 10% fetal bovine serum (Invitrogen, USA), Ellis salts, non-essential amino acids, sodium pyruvate, 100 μg / ml penicillin, and 10 units / ml streptomycin.

[0176] Cell penetration screening analysis using flow cytometry

[0177] To confirm the cell penetration of the peptides, Huh7 cell lines were treated with SEQ ID NO:2 to SEQ ID NO:178 and analyzed by flow cytometry. The analytical method was also confirmed using the method described in Example (1) for HeLa cells. The results of the analysis were... Figures 30 to 51 It is displayed in the middle.

[0178] (3) Cell penetration assay in human T lymphocyte lines

[0179] Cell culture

[0180] The Jurkat cell line (human T-cell leukemia cell line) was purchased from ATCC and used as a suspension cell line. The cells were cultured at 37°C in a 5% CO2 incubator in RPMI 1640 medium supplemented with 10% fetal bovine serum (Invitrogen, USA), Ellis salts, non-essential amino acids, sodium pyruvate, 100 μg / ml penicillin, and 10 units / ml streptomycin. Human-derived lymphocytes were isolated from healthy human blood (50 ml), and peripheral blood mononuclear cells (PBMCs) and lymphocytes were collected using Biocoll separation solution (Biochrom AG, Berlin, Germany).

[0181] Cell penetration screening analysis using flow cytometry

[0182] To confirm the cell penetration of the peptides, human T-cell lymphocyte lines were treated with SEQ ID NO:2 to SEQ ID NO:178 and analyzed by flow cytometry. The analytical method was also confirmed using the method described in Example (1) for HeLa cells. The results of the analysis were... Figures 52 to 69 It is displayed in the middle.

[0183] (4) Analysis of cell viability and cytotoxicity

[0184] HeLa cells were cultured in the same manner as in Example 4(1) above, aliquoted into 96-well plates, and cultured for 12 hours at 37°C in a 5% CO2 incubator in medium containing 10% fetal bovine serum (Invitrogen, USA), 100 μg / ml penicillin, and 10 units / ml streptomycin. After washing the cells with PBS, starvation was induced in minimum essential medium. Each carrier peptide was treated with 20 μM at 37°C and cultured for 1 hour. Cell viability and cytotoxicity were analyzed by MTT assay after cell culture. Results were presented in... Figures 70 to 86 It is displayed in the middle. <110> Jim Vickers & Kyle Ltd. Gold merchants <120> Cell-penetrating peptide, conjugates containing the peptide, and compositions containing the conjugates. <130> OF13P178 / CN <150> KR10‑2012‑0104173 <151> 2012‑09‑19 <150> KR10‑2012‑0104144 <151> 2012‑09‑19 <150> KR10‑2012‑0109207 <151> 2012‑09‑28 <150> KR10‑2012‑0109216 <151> 2012‑09‑28 <150> KR10‑2013‑0017046 <151> 2013‑02‑18 <160> 186 <170> PatentIn version 3.2 <210> 1 <211> 16 <212> PRT <213> Homo sapiens <400> 1 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 <210> 2 <211> 17 <212> PRT <213> Homo sapiens <400> 2 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys <210> 3 <211> 18 <212> PRT <213> Homo sapiens <400> 3 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys <210> 4 <211> 19 <212> PRT <213> Homo sapiens <400> 4 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys <210> 5 <211> 20 <212> PRT <213> Homo sapiens <400> 5 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys 20 <210> 6 <211> 21 <212> PRT <213> Homo sapiens <400> 6 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys 20 <210> 7 <211> 22 <212> PRT <213> Homo sapiens <400> 7 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys 20 <210> 8 <211> 23 <212> PRT <213> Homo sapiens <400> 8 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys 20 <210> 9 <211> 24 <212> PRT <213> Homo sapiens <400> 9 Glu Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr 1 5 10 15 Serum Arg Leu Arg Phe Ile Pro Lys 20 <210> 10 <211> 25 <212> PRT <213> Homo sapiens <400> 10 Ser Glu Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu 1 5 10 15 Thr Ser Arg Leu Arg Phe Ile Pro Lys 20 25 <210> 11 <211> 26 <212> PRT <213> Homo sapiens <400> 11 Leu Ser Glu Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu 1 5 10 15 Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 20 25 <210> 12 <211> 27 <212> PRT <213> Homo sapiens <400> 12 Glu Leu Ser Glu Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala 1 5 10 15 Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 20 25 <210> 13 <211> 28 <212> PRT <213> Homo sapiens <400> 13 Arg Glu Leu Ser Glu Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro 1 5 10 15 Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 20 25 <210> 14 <211> 29 <212> PRT <213> Homo sapiens <400> 14 Leu Arg Glu Leu Ser Glu Ala Glu Val Arg Gln His Arg Glu Ala Arg 1 5 10 15 Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 20 25 <210> 15 <211> 30 <212> PRT <213> Homo sapiens <400> 15 Gln Leu Arg Glu Leu Ser Glu Ala Glu Val Arg Gln His Arg Glu Ala 1 5 10 15 Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 20 25 30 <210> 16 <211> 18 <212> PRT <213> Homo sapiens <400> 16 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro <210> 17 <211> 19 <212> PRT <213> Homo sapiens <400> 17 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro <210> 18 <211> 20 <212> PRT <213> Homo sapiens <400> 18 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro Asp 20 <210> 19 <211> 19 <212> PRT <213> Homo sapiens <400> 19 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro Asp <210> 20 <211> 20 <212> PRT <213> Homo sapiens <400> 20 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Island Pro Lys Pro 20 <210> 21 <211> 21 <212> PRT <213> Homo sapiens <400> 21 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys Pro Asp 20 <210> 22 <211> 22 <212> PRT <213> Homo sapiens <400> 22 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys Pro Asp Gly 20 <210> 23 <211> 21 <212> PRT <213> Homo sapiens <400> 23 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro Asp Gly 20 <210> 24 <211> 20 <212> PRT <213> Homo sapiens <400> 24 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro Asp Gly 20 <210> 25 <211> 21 <212> PRT <213> Homo sapiens <400> 25 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro 20 <210> 26 <211> 22 <212> PRT <213> Homo sapiens <400> 26 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro Asp 20 <210> 27 <211> 23 <212> PRT <213> Homo sapiens <400> 27 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro Asp Gly 20 <210> 28 <211> 24 <212> PRT <213> Homo sapiens <400> 28 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro Asp Gly Leu 20 <210> 29 <211> 23 <212> PRT <213> Homo sapiens <400> 29 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys Pro Asp Gly Leu 20 <210> 30 <211> 22 <212> PRT <213> Homo sapiens <400> 30 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro Asp Gly Leu 20 <210> 31 <211> 21 <212> PRT <213> Homo sapiens <400> 31 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro Asp Gly Leu 20 <210> 32 <211> 22 <212> PRT <213> Homo sapiens <400> 32 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro 20 <210> 33 <211> 23 <212> PRT <213> Homo sapiens <400> 33 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro Asp 20 <210> 34 <211> 24 <212> PRT <213> Homo sapiens <400> 34 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro Asp Gly 20 <210> 35 <211> 25 <212> PRT <213> Homo sapiens <400> 35 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro Asp Gly Leu 20 25 <210> 36 <211> 26 <212> PRT <213> Homo sapiens <400> 36 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg 20 25 <210> 37 <211> 25 <212> PRT <213> Homo sapiens <400> 37 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro Asp Gly Leu Arg 20 25 <210> 38 <211> 24 <212> PRT <213> Homo sapiens <400> 38 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys Pro Asp Gly Leu Arg 20 <210> 39 <211> 23 <212> PRT <213> Homo sapiens <400> 39 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro Asp Gly Leu Arg 20 <210> 40 <211> 22 <212> PRT <213> Homo sapiens <400> 40 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro Asp Gly Leu Arg 20 <210> 41 <211> 23 <212> PRT <213> Homo sapiens <400> 41 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys Pro 20 <210> 42 <211> 24 <212> PRT <213> Homo sapiens <400> 42 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leo Arg Phe Ile Pro Lys Pro Asp 20 <210> 43 <211> 25 <212> PRT <213> Homo sapiens <400> 43 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys Pro Asp Gly 20 25 <210> 44 <211> 26 <212> PRT <213> Homo sapiens <400> 44 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu 20 25 <210> 45 <211> 27 <212> PRT <213> Homo sapiens <400> 45 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg 20 25 <210> 46 <211> 28 <212> PRT <213> Homo sapiens <400> 46 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro 20 25 <210> 47 <211> 27 <212> PRT <213> Homo sapiens <400> 47 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro 20 25 <210> 48 <211> 26 <212> PRT <213> Homo sapiens <400> 48 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro 20 25 <210> 49 <211> 25 <212> PRT <213> Homo sapiens <400> 49 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys Pro Asp Gly Leu Arg Pro 20 25 <210> 50 <211> 24 <212> PRT <213> Homo sapiens <400> 50 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro Asp Gly Leu Arg Pro 20 <210> 51 <211> 23 <212> PRT <213> Homo sapiens <400> 51 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro Asp Gly Leu Arg Pro 20 <210> 52 <211> 24 <212> PRT <213> Homo sapiens <400> 52 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro 20 <210> 53 <211> 25 <212> PRT <213> Homo sapiens <400> 53 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro Asp 20 25 <210> 54 <211> 26 <212> PRT <213> Homo sapiens <400> 54 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro Asp Gly 20 25 <210> 55 <211> 27 <212> PRT <213> Homo sapiens <400> 55 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu 20 25 <210> 56 <211> 28 <212> PRT <213> Homo sapiens <400> 56 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg 20 25 <210> 57 <211> 29 <212> PRT <213> Homo sapiens <400> 57 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro 20 25 <210> 58 <211> 30 <212> PRT <213> Homo sapiens <400> 58 Ala Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 10 15 Arg Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro Ile 20 25 30 <210> 59 <211> 29 <212> PRT <213> Homo sapiens <400> 59 Glu Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 15 Leu Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro Ile 20 25 <210> 60 <211> 28 <212> PRT <213> Homo sapiens <400> 60 Val Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 15 Arg Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro Ile 20 25 <210> 61 <211> 27 <212> PRT <213> Homo sapiens <400> 61 Arg Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 15 Phe Ile Pro Lys Pro Asp Gly Leu Arg Pro Ile 20 25 <210> 62 <211> 26 <212> PRT <213> Homo sapiens <400> 62 Gln His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 15 Ile Pro Lys Pro Asp Gly Leu Arg Pro Ile 20 25 <210> 63 <211> 25 <212> PRT <213> Homo sapiens <400> 63 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 15 Pro Lys Pro Asp Gly Leu Arg Pro Ile 20 25 <210> 64 <211> 24 <212> PRT <213> Homo sapiens <400> 64 Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 Lys Pro Asp Gly Leu Arg Pro Ile 20 <210> 65 <211> 17 <212> PRT <213> Homo sapiens <400> 65 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro <210> 66 <211> 18 <212> PRT <213> Homo sapiens <400> 66 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp <210> 67 <211> 19 <212> PRT <213> Homo sapiens <400> 67 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly <210> 68 <211> 21 <212> PRT <213> Homo sapiens <400> 68 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg 20 <210> 69 <211> 22 <212> PRT <213> Homo sapiens <400> 69 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro 20 <210> 70 <211> 23 <212> PRT <213> Homo sapiens <400> 70 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile 20 <210> 71 <211> 24 <212> PRT <213> Homo sapiens <400> 71 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val 20 <210> 72 <211> 25 <212> PRT <213> Homo sapiens <400> 72 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val Asn 20 25 <210> 73 <211> 26 <212> PRT <213> Homo sapiens <400> 73 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val Asn Met 20 25 <210> 74 <211> 27 <212> PRT <213> Homo sapiens <400> 74 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val Asn Met Asp 20 25 <210> 75 <211> 28 <212> PRT <213> Homo sapiens <400> 75 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val Asn Met Asp Tyr 20 25 <210> 76 <211> 29 <212> PRT <213> Homo sapiens <400> 76 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val Asn Met Asp Tyr Val 20 25 <210> 77 <211> 30 <212> PRT <213> Homo sapiens <400> 77 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 Pro Asp Gly Leu Arg Pro Ile Val Asn Met Asp Tyr Val Val 20 25 30 <210> 78 <211> 15 <212> PRT <213> Homo sapiens <400> 78 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 15 <210> 79 <211> 14 <212> PRT <213> Homo sapiens <400> 79 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 <210> 80 <211> 13 <212> PRT <213> Homo sapiens <400> 80 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 <210> 81 <211> 12 <212> PRT <213> Homo sapiens <400> 81 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 <210> 82 <211> 11 <212> PRT <213> Homo sapiens <400> 82 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 <210> 83 <211> 10 <212> PRT <213> Homo sapiens <400> 83 Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 10 <210> 84 <211> 9 <212> PRT <213> Homo sapiens <400> 84 Glu Ala Arg Pro Ala Leu Leu Thr Ser 1 5 <210> 85 <211> 8 <212> PRT <213> Homo sapiens <400> 85 Glu Ala Arg Pro Ala Leu Leu Thr 1 5 <210> 86 <211> 7 <212> PRT <213> Homo sapiens <400> 86 Glu Ala Arg Pro Ala Leu Leu 1 5 <210> 87 <211> 6 <212> PRT <213> Homo sapiens <400> 87 Glu Ala Arg Pro Ala Leu 1 5 <210> 88 <211> 5 <212> PRT <213> Homo sapiens <400> 88 Glu Ala Arg Pro Ala 1 5 <210> 89 <211> 4 <212> PRT <213> Homo sapiens <400> 89 Glu Ala Arg Pro 1 <210> 90 <211> 3 <212> PRT <213> Homo sapiens <400> 90 Glu Ala Arg 1 <210> 91 <211> 15 <212> PRT <213> Homo sapiens <400> 91 Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 15 <210> 92 <211> 14 <212> PRT <213> Homo sapiens <400> 92 Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 <210> 93 <211> 13 <212> PRT <213> Homo sapiens <400> 93 Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 <210> 94 <211> 12 <212> PRT <213> Homo sapiens <400> 94 Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 <210> 95 <211> 11 <212> PRT <213> Homo sapiens <400> 95 Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 <210> 96 <211> 10 <212> PRT <213> Homo sapiens <400> 96 Leu Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 10 <210> 97 <211> 9 <212> PRT <213> Homo sapiens <400> 97 Thr Ser Arg Leu Arg Phe Ile Pro Lys 1 5 <210> 98 <211> 8 <212> PRT <213> Homo sapiens <400> 98 Ser Arg Leu Arg Phe Ile Pro Lys 1 5 <210> 99 <211> 7 <212> PRT <213> Homo sapiens <400> 99 Arg Leu Arg Phe Ile Pro Lys 1 5 <210> 100 <211> 6 <212> PRT <213> Homo sapiens <400> 100 Leu Arg Phe Ile Pro Lys 1 5 <210> 101 <211> 5 <212> PRT <213> Homo sapiens <400> 101 Arg Phe Ile Pro Lys 1 5 <210> 102 <211> 4 <212> PRT <213> Homo sapiens <400> 102 Phe Ile Pro Lys 1 <210> 103 <211> 3 <212> PRT <213> Homo sapiens <400> 103 Ile Pro Lys 1 <210> 104 <211> 14 <212> PRT <213> Homo sapiens <400> 104 Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 <210> 105 <211> 12 <212> PRT <213> Homo sapiens <400> 105 Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 <210> 106 <211> 10 <212> PRT <213> Homo sapiens <400> 106 Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 <210> 107 <211> 8 <212> PRT <213> Homo sapiens <400> 107 Ala Leu Leu Thr Ser Arg Leu Arg 1 5 <210> 108 <211> 6 <212> PRT <213> Homo sapiens <400> 108 Leu Leu Thr Ser Arg Leu 1 5 <210> 109 <211> 4 <212> PRT <213> Homo sapiens <400> 109 Leu Thr Ser Arg 1 <210> 110 <211> 13 <212> PRT <213> Homo sapiens <400> 110 Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 <210> 111 <211> 12 <212> PRT <213> Homo sapiens <400> 111 Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 <210> 112 <211> 11 <212> PRT <213> Homo sapiens <400> 112 Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 <210> 113 <211> 10 <212> PRT <213> Homo sapiens <400> 113 Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu 1 5 10 <210> 114 <211> 9 <212> PRT <213> Homo sapiens <400> 114 Ala Arg Pro Ala Leu Leu Thr Ser Arg 1 5 <210> 115 <211> 8 <212> PRT <213> Homo sapiens <400> 115 Ala Arg Pro Ala Leu Leu Thr Ser 1 5 <210> 116 <211> 7 <212> PRT <213> Homo sapiens <400> 116 Ala Arg Pro Ala Leu Leu Thr 1 5 <210> 117 <211> 6 <212> PRT <213> Homo sapiens <400> 117 Arg Wing Pro Leu Wing Leu 1 5 <210> 118 <211> 5 <212> PRT <213> Homo sapiens <400> 118 Arg Wing Pro Leo Wing 1 5 <210> 119 <211> 4 <212> PRT <213> Homo sapiens <400> 119 Ala Arg Pro Ala 1 <210> 120 <211> 3 <212> PRT <213> Homo sapiens <400> 120 Arg Pro Wing 1 <210> 121 <211> 13 <212> PRT <213> Homo sapiens <400> 121 Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 <210> 122 <211> 11 <212> PRT <213> Homo sapiens <400> 122 Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 10 <210> 123 <211> 10 <212> PRT <213> Homo sapiens <400> 123 Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 10 <210> 124 <211> 8 <212> PRT <213> Homo sapiens <400> 124 Arg Pro Ala Leu Leu Thr Ser Arg 1 5 <210> 125 <211> 7 <212> PRT <213> Homo sapiens <400> 125 Arg Pro Ala Leu Leu Thr Ser 1 5 <210> 126 <211> 6 <212> PRT <213> Homo sapiens <400> 126 Arg Pro Ala Leu Leu Thr 1 5 <210> 127 <211> 5 <212> PRT <213> Homo sapiens <400> 127 Arg Pro Ala Leo Leo 1 5 <210> 128 <211> 4 <212> PRT <213> Homo sapiens <400> 128 Arg Pro Wing Leo 1 <210> 129 <211> 3 <212> PRT <213> Homo sapiens <400> 129 Arg Pro Ala 1 <210> 130 <211> 12 <212> PRT <213> Homo sapiens <400> 130 Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 <210> 131 <211> 11 <212> PRT <213> Homo sapiens <400> 131 Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 10 <210> 132 <211> 9 <212> PRT <213> Homo sapiens <400> 132 Pro Ala Leu Leu Thr Ser Arg Leu Arg 1 5 <210> 133 <211> 8 <212> PRT <213> Homo sapiens <400> 133 Pro Ala Leu Leu Thr Ser Arg Leu 1 5 <210> 134 <211> 7 <212> PRT <213> Homo sapiens <400> 134 Pro Ala Leu Leu Thr Ser Arg 1 5 <210> 135 <211> 6 <212> PRT <213> Homo sapiens <400> 135 Pro Ala Leu Leu Thr Ser 1 5 <210> 136 <211> 5 <212> PRT <213> Homo sapiens <400> 136 Pro Ala Leo Leo Thr 1 5 <210> 137 <211> 4 <212> PRT <213> Homo sapiens <400> 137 Pro Wing Leo Leo 1 <210> 138 <211> 3 <212> PRT <213> Homo sapiens <400> 138 Pro Wing Leo 1 <210> 139 <211> 11 <212> PRT <213> Homo sapiens <400> 139 Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 <210> 140 <211> 9 <212> PRT <213> Homo sapiens <400> 140 Ala Leu Leu Thr Ser Arg Leu Arg Phe 1 5 <210> 141 <211> 7 <212> PRT <213> Homo sapiens <400> 141 Ala Leu Leu Thr Ser Arg Leu 1 5 <210> 142 <211> 6 <212> PRT <213> Homo sapiens <400> 142 Ala Leu Leu Thr Ser Arg 1 5 <210> 143 <211> 5 <212> PRT <213> Homo sapiens <400> 143 Ala Leu Leu Thr Ser 1 5 <210> 144 <211> 4 <212> PRT <213> Homo sapiens <400> 144 Wing Leo Leo Thr 1 <210> 145 <211> 3 <212> PRT <213> Homo sapiens <400> 145 Ala Leo Leo 1 <210> 146 <211> 10 <212> PRT <213> Homo sapiens <400> 146 Leu Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 10 <210> 147 <211> 9 <212> PRT <213> Homo sapiens <400> 147 Leu Leu Thr Ser Arg Leu Arg Phe Ile 1 5 <210> 148 <211> 7 <212> PRT <213> Homo sapiens <400> 148 Leu Leu Thr Ser Arg Leu Arg 1 5 <210> 149 <211> 5 <212> PRT <213> Homo sapiens <400> 149 Leu Leu Thr Ser Arg 1 5 <210> 150 <211> 4 <212> PRT <213> Homo sapiens <400> 150 Leo Leo Thr Ser 1 <210> 151 <211> 3 <212> PRT <213> Homo sapiens <400> 151 Leo Leo Thr 1 <210> 152 <211> 9 <212> PRT <213> Homo sapiens <400> 152 Leu Thr Ser Arg Leu Arg Phe Ile Pro 1 5 <210> 153 <211> 8 <212> PRT <213> Homo sapiens <400> 153 Leu Thr Ser Arg Leu Arg Phe Ile 1 5 <210> 154 <211> 7 <212> PRT <213> Homo sapiens <400> 154 Leu Thr Ser Arg Leu Arg Phe 1 5 <210> 155 <211> 6 <212> PRT <213> Homo sapiens <400> 155 Leu Thr Ser Arg Leu Arg 1 5 <210> 156 <211> 5 <212> PRT <213> Homo sapiens <400> 156 Leu Thr Ser Arg Leu 1 5 <210> 157 <211> 3 <212> PRT <213> Homo sapiens <400> 157 Leu Thr Ser 1 <210> 158 <211> 8 <212> PRT <213> Homo sapiens <400> 158 Thr Ser Arg Leu Arg Phe Ile Pro 1 5 <210> 159 <211> 7 <212> PRT <213> Homo sapiens <400> 159 Thr Ser Arg Leu Arg Phe Ile 1 5 <210> 160 <211> 6 <212> PRT <213> Homo sapiens <400> 160 Thr Ser Arg Leu Arg Phe 1 5 <210> 161 <211> 5 <212> PRT <213> Homo sapiens <400> 161 Thr Ser Arg Leu Arg 1 5 <210> 162 <211> 4 <212> PRT <213> Homo sapiens <400> 162 Thr Ser Arg Leu 1 <210> 163 <211> 3 <212> PRT <213> Homo sapiens <400> 163 Thr Ser Arg 1 <210> 164 <211> 7 <212> PRT <213> Homo sapiens <400> 164 Ser Arg Leu Arg Phe Ile Pro 1 5 <210> 165 <211> 6 <212> PRT <213> Homo sapiens <400> 165 Ser Arg Leu Arg Phe Ile 1 5 <210> 166 <211> 5 <212> PRT <213> Homo sapiens <400> 166 Ser Arg Leu Arg Phe 1 5 <210> 167 <211> 4 <212> PRT <213> Homo sapiens <400> 167 Ser Arg Leu Arg 1 <210> 168 <211> 3 <212> PRT <213> Homo sapiens <400> 168 Ser Arg Leu 1 <210> 169 <211> 6 <212> PRT <213> Homo sapiens <400> 169 Arg Leu Arg Phe Ile Pro 1 5 <210> 170 <211> 5 <212> PRT <213> Homo sapiens <400> 170 Arg Leu Arg Phe Ile 1 5 <210> 171 <211> 4 <212> PRT <213> Homo sapiens <400> 171 Arg Leu Arg Phe 1 <210> 172 <211> 3 <212> PRT <213> Homo sapiens <400> 172 Arg Leu Arg 1 <210> 173 <211> 5 <212> PRT <213> Homo sapiens <400> 173 Leu Arg Phe Ile Pro 1 5 <210> 174 <211> 4 <212> PRT <213> Homo sapiens <400> 174 Leu Arg Phe Ile 1 <210> 175 <211> 3 <212> PRT <213> Homo sapiens <400> 175 Leu Arg Phe 1 <210> 176 <211> 4 <212> PRT <213> Homo sapiens <400> 176 Arg Phe Ile Pro 1 <210> 177 <211> 3 <212> PRT <213> Homo sapiens <400> 177 Arg Phe Ile 1 <210> 178 <211> 3 <212> PRT <213> Homo sapiens <400> 178 Phe Ile Pro 1 <210> 179 <211> 1132 <212> PRT <213> Homo sapiens <400> 179 Met Pro Arg Ala Pro Arg Cys Arg Ala Val Arg Ser Leu Leu Arg Ser 1 5 10 15 His Tyr Arg Glu Val Leu Pro Leu Ala Thr Phe Val Arg Arg Leu Gly 20 25 30 Pro Gln Gly Trp Arg Leu Val Gln Arg Gly Asp Pro Ala Ala Phe Arg 35 40 45 Ala Leu Val Ala Gln Cys Leu Val Cys Val Pro Trp Asp Ala Arg Pro 50 55 60 Pro Pro Ala Ala Pro Ser Phe Arg Gln Val Ser Cys Leu Lys Glu Leu 65 70 75 80 Val Ala Arg Val Leu Gln Arg Leu Cys Glu Arg Gly Ala Lys Asn Val 85 90 95 Leu Ala Phe Gly Phe Ala Leu Leu Asp Gly Ala Arg Gly Gly Pro Pro 100 105 110 Glu Ala Phe Thr Thr Ser Val Arg Ser Tyr Leu Pro Asn Thr Val Thr 115 120 125 Asp Ala Leu Arg Gly Ser Gly Ala Trp Gly Leu Leu Leu Arg Arg Val 130 135 140 Gly Asp Asp Val Leu Val His Leu Leu Ala Arg Cys Ala Leu Phe Val 145 150 155 160 Leu Val Ala Pro Ser Cys Ala Tyr Gln Val Cys Gly Pro Pro Leu Tyr 165 170 175 Gln Leu Gly Ala Ala Thr Gln Ala Arg Pro Pro Pro His Ala Ser Gly 180 185 190 Pro Arg Arg Arg Leu Gly Cys Glu Arg Ala Trp Asn His Ser Val Arg 195 200 205 Glu Ala Gly Val Pro Leu Gly Leu Pro Ala Pro Gly Ala Arg Arg Arg 210 215 220 Gly Gly Ser Ala Ser Arg Ser Leu Pro Leu Pro Lys Arg Pro Arg Arg 225 230 235 240 Gly Ala Ala Pro Glu Pro Glu Arg Thr Pro Val Gly Gln Gly Ser Trp 245 250 255 Ala His Pro Gly Arg Thr Arg Gly Pro Ser Asp Arg Gly Phe Cys Val 260 265 270 Val Ser Pro Ala Arg Pro Ala Glu Glu Ala Thr Ser Leu Glu Gly Ala 275 280 285 Leu Ser Gly Thr Arg His Ser His Pro Ser Val Gly Arg Gln His His 290 295 300 Ala Gly Pro Pro Ser Thr Ser Arg Pro Pro Arg Pro Trp Asp Thr Pro 305 310 315 320 Cys Pro Pro Val Tyr Ala Glu Thr Lys His Phe Leu Tyr Ser Ser Gly 325 330 335 Asp Lys Glu Gln Leu Arg Pro Ser Phe Leu Leu Ser Ser Leu Arg Pro 340 345 350 Ser Leu Thr Gly Ala Arg Arg Leu Val Glu Thr Ile Phe Leu Gly Ser 355 360 365 Arg Pro Trp Met Pro Gly Thr Pro Arg Arg Leu Pro Arg Leu Pro Gln 370 375 380 Arg Tyr Trp Gln Met Arg Pro Leu Phe Leu Glu Leu Leu Gly Asn His 385 390 395 400 Ala Gln Cys Pro Tyr Gly Val Leu Leu Lys Thr His Cys Pro Leu Arg 405 410 415 Ala Ala Val Thr Pro Ala Ala Gly Val Cys Ala Arg Glu Lys Pro Gln 420 425 430 Gly Ser Val Ala Ala Pro Glu Glu Glu Asp Thr Asp Pro Arg Arg Leu 435 440 445 Val Gln Leu Leu Arg Gln His Ser Ser Pro Trp Gln Val Tyr Gly Phe 450 455 460 Val Arg Ala Cys Leu Arg Arg Leu Val Pro Pro Gly Leu Trp Gly Ser 465 470 475 480 Arg His Asn Glu Arg Arg Phe Leu Arg Asn Thr Lys Lys Phe Ile Ser 485 490 495 Leu Gly Lys His Ala Lys Leu Ser Leu Gln Glu Leu Thr Trp Lys Met 500 505 510 Ser Val Arg Asp Cys Ala Trp Leu Arg Arg Ser Pro Gly Val Gly Cys 515 520 525 Val Pro Ala Ala Glu His Arg Leu Arg Glu Glu Ile Leu Ala Lys Phe 530 535 540 Leu His Trp Leu Met Ser Val Tyr Val Val Glu Leu Leu Arg Ser Phe 545 550 555 560 Phe Tyr Val Thr Glu Thr Thr Phe Gln Lys Asn Arg Leu Phe Phe Tyr 565 570 575 Arg Lys Ser Val Trp Ser Lys Leu Gln Ser Ile Gly Ile Arg Gln His 580 585 590 Leu Lys Arg Val Gln Leu Arg Glu Leu Ser Glu Ala Glu Val Arg Gln 595 600 605 His Arg Glu Ala Arg Pro Ala Leu Leu Thr Ser Arg Leu Arg Phe Ile 610 615 620 Pro Lys Pro Asp Gly Leu Arg Pro Ile Val Asn Met Asp Tyr Val Val 625 630 635 640 Gly Ala Arg Thr Phe Arg Arg Glu Lys Arg Ala Glu Arg Leu Thr Ser 645 650 655 Arg Val Lys Ala Leu Phe Ser Val Leu Asn Tyr Glu Arg Ala Arg Arg 660 665 670 Pro Gly Leu Leu Gly Ala Ser Val Leu Gly Leu Asp Asp Ile His Arg 675 680 685 Ala Trp Arg Thr Phe Val Leu Arg Val Arg Ala Gln Asp Pro Pro Pro 690 695 700 Glu Leu Tyr Phe Val Lys Val Asp Val Thr Gly Ala Tyr Asp Thr Ile 705 710 715 720 Pro Gln Asp Arg Leu Thr Glu Val Ile Ala Ser Ile Ile Lys Pro Gln 725 730 735 Asn Thr Tyr Cys Val Arg Arg Tyr Ala Val Val Gln Lys Ala Ala His 740 745 750 Gly His Val Arg Lys Ala Phe Lys Ser His Val Ser Thr Leu Thr Asp 755 760 765 Leu Gln Pro Tyr Met Arg Gln Phe Val Ala His Leu Gln Glu Thr Ser 770 775 780 Pro Leu Arg Asp Ala Val Val Ile Glu Gln Ser Ser Ser Leu Asn Glu 785 790 795 800 Ala Ser Ser Gly Leu Phe Asp Val Phe Leu Arg Phe Met Cys His His 805 810 815 Ala Val Arg Ile Arg Gly Lys Ser Tyr Val Gln Cys Gln Gly Ile Pro 820 825 830 Gln Gly Ser Ile Leu Ser Thr Leu Leu Cys Ser Leu Cys Tyr Gly Asp 835 840 845 Met Glu Asn Lys Leu Phe Ala Gly Ile Arg Arg Asp Gly Leu Leu Leu 850 855 860 Arg Leu Val Asp Asp Phe Leu Leu Val Thr Pro His Leu Thr His Ala 865 870 875 880 Lys Thr Phe Leu Arg Thr Leu Val Arg Gly Val Pro Glu Tyr Gly Cys 885 890 895 Val Val Asn Leu Arg Lys Thr Val Val Asn Phe Pro Val Glu Asp Glu 900 905 910 Ala Leu Gly Gly Thr Ala Phe Val Gln Met Pro Ala His Gly Leu Phe 915 920 925 Pro Trp Cys Gly Leu Leu Leu Asp Thr Arg Thr Leu Glu Val Gln Ser 930 935 940 Asp Tyr Ser Ser Tyr Ala Arg Thr Ser Ile Arg Ala Ser Leu Thr Phe 945 950 955 960 Asn Arg Gly Phe Lys Ala Gly Arg Asn Met Arg Arg Lys Leu Phe Gly 965 970 975 Val Leu Arg Leu Lys Cys His Ser Leu Phe Leu Asp Leu Gln Val Asn 980 985 990 Ser Leu Gln Thr Val Cys Thr Asn Ile Tyr Lys Ile Leu Leu Leu Gln 995 1000 1005 Ala Tyr Arg Phe His Ala Cys Val Leu Gln Leu Pro Phe His Gln Gln 1010 1015 1020 Val Trp Lys Asn Pro Thr Phe Phe Leu Arg Val Ile Ser Asp Thr Ala 1025 1030 1035 1040 Ser Leu Cys Tyr Ser Ile Leu Lys Ala Lys Asn Ala Gly Met Ser Leu 1045 1050 1055 Gly Ala Lys Gly Ala Ala Gly Pro Leu Pro Ser Glu Ala Val Gln Trp 1060 1065 1070 Leu Cys His Gln Ala Phe Leu Leu Lys Leu Thr Arg His Arg Val Thr 1075 1080 1085 Tyr Val Pro Leu Leu Gly Ser Leu Arg Thr Ala Gln Thr Gln Leu Ser 1090 1095 1100 Arg Lys Leu Pro Gly Thr Thr Leu Thr Ala Leu Glu Ala Ala Ala Asn 1105 1110 1115 1120 Pro Ala Leu Pro Ser Asp Phe Lys Thr Ile Leu Asp 1125 1130 <210> 180 <211> 48 <212> DNA <213> Homo sapiens <400> 180 gaagcgcgcc cggcgctgct gaccagccgc ctgcgcttta ttccgaaa 48 <210> 181 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> siRNA sense <220> <221> variation <222> (20)..(21) <223> n is deoxythymidine (dT) <400> 181 cuuacgcuga guacuucgan n 21 <210> 182 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> siRNA antisense <220> <221> variation <222> (20)..(21) <223> n is deoxythymidine (dT) <400> 182 ucgaaguacu cagcguaagn n 21 <210> 183 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> Green Fluorescent Protein <400> 183 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 184 <211> 720 <212> DNA <213> Artificial Sequence <220> <223> Green Fluorescent Protein <400> 184 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 60 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac 120 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 180 cctgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 240 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctttc 300 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 360 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 420 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 480 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 540 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 600 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 660 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa 720 720 <210> 185 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> siCont sense <220> <221> variation <222> (20)..(21) <223> n is deoxythymidine (dT) <400> 185 gcaccuauaa caacgguagn n 21 <210> 186 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> siCont antisense <220> <221> variation <222> (20)..(21) <223> n is deoxythymidine (dT) <400> 186 cuaccguugu uauaggugcn n 21

Claims

1. The use of a cell-penetrating carrier peptide in the preparation of a composition comprising the cell-penetrating carrier peptide and a cargo to be transported, wherein, The cell-penetrating carrier peptide delivers the cargo into the cell, and The cell-penetrating carrier peptide is a peptide composed of any amino acid sequence selected from the group consisting of SEQ ID NO:111 to SEQ ID NO:113 and SEQ ID NO:115 to SEQ ID NO:

120. The goods in question are contrast agents.

2. The application as described in claim 1, wherein, The carrier peptide is directly coupled to the cargo via covalent bonds or indirectly coupled to the cargo via a connector via covalent bonds.

3. The application as described in claim 1, wherein, The carrier peptide and the cargo are coupled by non-covalent bonds.

4. The application as described in claim 1, wherein, The contrast agent is selected from the group consisting of radiopaque contrast agents, paramagnetic contrast agents, superparamagnetic contrast agents, and CT contrast agents.

5. The application as described in claim 1, wherein, The contrast agent is iron-based.

6. The application as described in claim 5, wherein, The contrast agent is ferrocene carboxylate.

7. The application as described in claim 1, wherein the composition is a contrast agent.

8. The application as described in claim 7, wherein, The contrast agent is used to contrast the cells.

9. The application as described in claim 8, wherein, The cells in question are stem cells.

10. The application as described in claim 1, wherein, The composition is a pharmaceutical composition used to treat or prevent disease.

11. The application as described in claim 1, wherein, The composition is a cosmetic composition.

12. The application as described in claim 1, wherein, The composition is a health food composition.

13. A cell-penetrating peptide, wherein, The cell-penetrating peptide is composed of any amino acid sequence selected from the group consisting of SEQ ID NO:111 to SEQ ID NO:113 and SEQ ID NO:115 to SEQ ID NO:

120.

14. A polynucleotide encoding the cell-penetrating peptide of claim 13.

15. A vector comprising the polynucleotide of claim 14.

16. A transformed cell comprising the vector of claim 15.

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