WO2016080633A1 - Peptides pour le ciblage de cancer gastrique, et leur utilisation médicale - Google Patents

Peptides pour le ciblage de cancer gastrique, et leur utilisation médicale Download PDF

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WO2016080633A1
WO2016080633A1 PCT/KR2015/007943 KR2015007943W WO2016080633A1 WO 2016080633 A1 WO2016080633 A1 WO 2016080633A1 KR 2015007943 W KR2015007943 W KR 2015007943W WO 2016080633 A1 WO2016080633 A1 WO 2016080633A1
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peptide
gastric cancer
cancer
targeting
peptides
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PCT/KR2015/007943
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English (en)
Korean (ko)
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최은경
정성윤
송시열
이경진
신설화
주은진
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울산대학교 산학협력단
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Priority claimed from KR1020150106581A external-priority patent/KR101693431B1/ko
Application filed by 울산대학교 산학협력단 filed Critical 울산대학교 산학협력단
Priority to US15/527,717 priority Critical patent/US20170322213A1/en
Priority to EP15860565.9A priority patent/EP3223016B1/fr
Publication of WO2016080633A1 publication Critical patent/WO2016080633A1/fr
Priority to US16/149,142 priority patent/US10627402B2/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer

Definitions

  • the present invention relates to a stomach cancer target peptide and gastric cancer diagnostic composition and drug delivery according to the radiation irradiation using the same.
  • cancer The smallest unit of the human body, the cell, normally divides, grows and dies by intracellular control and maintains cell balance. If a cell is damaged for some reason, it is treated and recovered to act as a normal cell, but if it is not recovered, it dies on its own.
  • cancer is defined as a condition in which abnormal cells, which are not controlled for such proliferation and suppression, proliferate excessively, invade surrounding tissues and organs, and cause mass formation and destruction of normal tissues for various reasons. Cancer is the proliferation of non-suppressive cells, which destroys the structure and function of normal cells and organs, so the diagnosis and treatment are of great importance.
  • drug delivery systems or targeted therapies that selectively deliver drugs to cancer cells and tissues are techniques of great interest. Because using the same amount of anticancer drugs can increase the efficacy of the drug and at the same time significantly reduce side effects on normal tissues.
  • the virus when applied to gene therapy, the virus can be selectively delivered to cancer cells to increase treatment efficiency and reduce serious side effects. To this end, it has been mainly developed antigens specific to tumor cells and antibodies targeting them. However, in the case of antibodies, there are problems such as fear of immune response and low efficiency of penetration into tissues. Peptides, on the other hand, have a low molecular weight, which is less likely to cause an immune response and is easier to penetrate into tissues. Therefore, when the cancer target peptide is connected with an existing anticancer agent, it can be utilized as an intelligent drug delivery agent that selectively delivers drugs to tumors.
  • the present invention constructs a mouse model in which actual human cancer tissue is transplanted, and divides it into a group of irradiated population and a group not irradiated with the control group, and is specific to each population.
  • a mouse model in which actual human cancer tissue is transplanted, and divides it into a group of irradiated population and a group not irradiated with the control group, and is specific to each population.
  • the present invention provides a peptide for gastric cancer target consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 6 and a polynucleotide encoding the same.
  • the present invention provides a peptide for gastric cancer targeting consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 3 specific to the gastric cancer tissues irradiated.
  • the present invention provides a drug delivery composition comprising the peptide for gastric cancer targeting.
  • the present invention relates to a stomach cancer target peptide and gastric cancer diagnostic composition and drug delivery according to the irradiation using the same, the treatment responsiveness due to the genetic differences of individual cancer patients with different treatment cases according to different treatment cases
  • functional peptides that can be targeted to implement customized diagnosis and treatment for each patient.
  • An animal model similar to a cancer microenvironment of an actual patient was constructed and divided into a group irradiated and a group unirradiated with its control group, and screened peptides specifically binding to each population to target each of the selected peptides. The efficiency was verified. Through this, it can be used to develop an image diagnosis technology for predicting the responsiveness of radiotherapy and to develop a customized target therapy accordingly.
  • A Gastric cancer tissue from BRC, B: Xenotransplantation into NOD / SCID mouse flank; C: B cancer tissue grows to 400-500 mm 3 , then cancer tissue 3x3 mm for passage Amputation, D: Mouse model in which anesthesia is injected into the nude mouse and one xenograft is transplanted to both subcutaneous subcutaneous tissues. E: A wide area where cancer tissue is formed when cancer tissue grows to a size of 150-200 mm 3 . The radiation dose of 10Gy was irradiated only on the leg part. The control group was not irradiated.
  • FIG. 2 is a biopanning scheme for identifying peptide sequences targeting in vivo patient gastric cancer tissues irradiated using the M13 phage display method according to an embodiment of the present invention.
  • Figure 3 is a result of comparing the phage concentration by extracting the phage after extraction of the heart, lung, liver, spleen, kidney, tumor during the five biopanning process.
  • FIG. 5 shows the results of confirming that the peptide phages connected to Cy5.5 fluorescence were respectively injected into the mouse model, and then specifically confirmed by imaging on the second day.
  • FIG. 6 shows the results of clarifying the fluorescence intensity of only cancer tissues and equally dividing the cancer tissues by extracting only cancer tissues and ending each day in vivo imaging.
  • Figure 7 is a plan for observing the change in target capacity of the peptide phage selected in the present invention, with or without irradiation.
  • Mouse model grown to 150-200 mm 3 after gastric cancer tissue transplantation 1)
  • Non-irradiated mouse model 2 10Gy irradiated mouse model, divided into two groups, 10Gy irradiated mice within 24 hours recovery time After having, the selected peptide phage sample was injected and confirmed in vivo imaging until the second day.
  • FIG. 10 is the result of connecting the peptide to the drug-encapsulated liposomes.
  • FIG. 10A Schematic diagram of connecting peptides to drug-encapsulated liposomes and chemical structural formulas expressing actual linking residues.
  • FIG. 10B Reduction test results for calculating how much -SH residues are in liposomes before peptide linkage and size distribution after peptide linkage. This is the result of checking stability through verification.
  • FIG. 13 shows the results of confirming the selection peptide targeting ability upon irradiation to another patient gastric cancer tissue transplant mouse according to Example 8.
  • the present invention provides a peptide for gastric cancer targeting consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 6.
  • the amino acid sequence of this SEQ ID NO is listed in Table 1.
  • the peptide of the present invention is a low molecular peptide consisting of seven amino acids.
  • Small molecule peptides have the advantage of being small in size, stabilized in three dimensions, easily passing through mucous membranes, and recognizing molecular targets deep in tissues.
  • the small molecule peptide according to the present invention has the advantage of early diagnosis of cancer because stability can be secured through local injection and minimize the immune response.
  • the low molecular weight peptides according to the present invention are relatively simple in mass production and weak in toxicity.
  • the low molecular weight peptide according to the present invention has an advantage of having a higher binding strength to the target material than the antibody, and does not occur degeneration during thermal / chemical treatment.
  • the small size of the molecule can be used as a fusion protein by attaching to other proteins. Specifically, since it can be used by attaching to a polymer protein chain, it can be used as a diagnostic kit and drug delivery material.
  • Peptides of the present invention can be readily prepared by chemical synthesis known in the art (Creighton, Proteins; Structures and Molecular Principles, W. H. Freeman and Co., NY, 1983). Representative methods include liquid or solid phase synthesis, fragment condensation, F-MOC or T-BOC chemistry (Chemical Approaches to the Synthesis of Peptides and Proteins, Williams et al., Eds., CRC Press, Boca Raton Florida, 1997 A Practical Approach, Athert on & Sheppard, Eds., IRL Press, Oxford, England, 1989).
  • the peptide of the present invention can be prepared by genetic engineering methods.
  • a DNA sequence encoding the peptide is constructed according to a conventional method.
  • DNA sequences can be constructed by PCR amplification using appropriate primers.
  • DNA sequences may be synthesized by standard methods known in the art, such as using automated DNA synthesizers (such as those sold by Biosearch or AppliedBiosystems).
  • the constructed DNA sequence is a vector comprising one or more expression control sequences (e.g., promoters, enhancers, etc.) that are operatively linked to this DNA sequence to regulate expression of the DNA sequence.
  • the host cell is transformed with the recombinant expression vector formed therefrom.
  • substantially pure peptide means that the peptide according to the invention is substantially free of any other protein derived from the host.
  • the present invention provides a peptide for gastric cancer targeting consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 3 specific to the gastric cancer tissues irradiated.
  • the peptide consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 3 of the present invention specifically binds to gastric cancer tissues, especially gastric cancer tissues irradiated.
  • target or “specific” means the ability to specifically bind to stomach cancer tissues, in particular radiation-treated stomach cancer tissues without binding to other normal tissues.
  • the gastric cancer specific peptide may specifically bind inside or outside gastric cancer tissue.
  • the present invention also provides a polynucleotide encoding any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 3.
  • polynucleotide is a polymer of deoxyribonucleotides or ribonucleotides present in single- or double-stranded form. It encompasses RNA genomic sequences, DNA (gDNA and cDNA) and RNA sequences transcribed therefrom and includes analogs of natural polynucleotides unless specifically stated otherwise.
  • the polynucleotide includes not only the nucleotide sequence encoding the gastric cancer target peptide, but also a sequence complementary to the sequence.
  • Such complementary sequences include sequences that are substantially complementary, as well as sequences that are substantially complementary.
  • the polynucleotide may also be modified. Such modifications include addition, deletion or non-conservative substitutions or conservative substitutions of nucleotides.
  • the polynucleotide encoding the amino acid sequence is to be interpreted to also include a nucleotide sequence showing substantial identity to the nucleotide sequence. The substantial identity is at least 80% homology when the nucleotide sequence is aligned with any other sequence as closely as possible and the aligned sequence is analyzed using algorithms commonly used in the art. A sequence exhibiting at least 90% homology or at least 95% homology.
  • the present invention also provides a gastric cancer diagnostic composition
  • a gastric cancer diagnostic composition comprising a peptide for gastric cancer target consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 6.
  • the present invention also provides a radiation reactive gastric cancer diagnostic composition consisting of any one amino acid sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 3 comprising a gastric cancer targeting peptide specific to the radiation-treated gastric cancer tissue. .
  • diagnosis in the present invention is meant identifying the presence or characteristic of a pathological condition.
  • diagnosis is to identify the presence or characteristics of gastric cancer.
  • Diagnosis of gastric cancer using the peptide of the present invention can be diagnosed by reacting the peptide of the present invention with a tissue or cell obtained directly by blood, urine or biopsy to detect their binding.
  • the peptide of the present invention may be provided in a labeled state.
  • the detectable label may be provided in a link (eg, covalently bonded or crosslinked).
  • the detectable label may be a colorase (e.g. peroxidase, alkaline phosphatase), radioisotope (e.g. 124 I, 125 I, 111 In, 99 mTc, 32 P, 35 S), Chromophores, luminescent or fluorescent materials (e.g. FITC, RITC, rhodamine, cyanine, Texas red, fluorescein, phycoerythrin, And quantum dots.
  • the detectable label can be an antibody epitope, substrate, cofactor, inhibitor or affinity ligand. Such labeling may be carried out in the course of synthesizing the peptide of the present invention, or may be performed in addition to the peptide already synthesized.
  • fluorescent material is used as a detectable label
  • cancer can be diagnosed by fluorescence mediated tomography (FMT).
  • FMT fluorescence mediated tomography
  • the peptide of the present invention labeled with a fluorescent material can be circulated into the blood and fluorescence by the peptide can be observed by fluorescence tomography. If fluorescence is observed, it is diagnosed as cancer.
  • the present invention provides a drug delivery composition comprising the peptide for gastric cancer targeting.
  • Peptides according to the present invention can be used as an intelligent drug carrier for selectively delivering drugs to cancer tissue. If the peptide of the present invention is used in the treatment of cancer in connection with a conventionally known drug, since the drug is selectively delivered only to cancer tissue and cancer cells by the peptide of the present invention, the effect of the drug can be increased and at the same time, the drug affects the normal tissue. Side effects can be significantly reduced.
  • the drug is an anticancer agent, and any anticancer agent that may be linked to the peptide of the present invention may be used without limitation as long as it is used in the conventional treatment of cancer.
  • anticancer agent examples include cisplatin, 5-fluorouracil, adriamycin, methotrexate, vinblastine, busulfan, chlorambucil, cyclophosphamide, melphalan, nitrogen mustard, nitrosourea, taxol, paclitaxel, docetaxel, 6 Mercaptopurine, 6-thioguanine, bleomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitomycin-C, hydroxyurea and the like.
  • Linkage between the anticancer agent and the peptide of the present invention can be carried out through methods known in the art, such as covalent bonds, crosslinking and the like.
  • the peptide of the present invention may be chemically modified if necessary so long as its activity is not lost.
  • anesthesia was intraperitoneally injected into NOD / SCID mice, followed by anesthesia, cutting the cancer tissue to a size of 3x3 mm, and transplanting one tissue to each side of the mouse subcutaneously, 100 ⁇ l of 100 IU / ml penicillin, 100 ⁇ g / After treatment with ml streptomycin, 100 ⁇ g / ml gentamicin and 2.5 ⁇ g / ml amphotericin B antibiotic solution, suture and recover on a warm pad (approximately 2 hours). Thereafter, cancer tissue formation and growth is observed every week, and if the cancer tissue grows to a size of 400-500 mm 3 , the cancer tissue is separated and cut into 3 ⁇ 3 mm sizes for passage culture.
  • the library is a loop-type peptide library [library fused to M13 phage gp3 minor coat protein] -Ph.D TM phage display peptide library, which has about 2.7 billion diverse amino acid sequences through a random arrangement of 7 amino acids.
  • kit, New England Biolabs (NEB) was purchased and used.
  • M13 phage peptide library (a loop peptide consisting of 7 amino acids with approximately 2.7 billion different amino acid sequences) was passed through the tail vein of the mouse.
  • M13 phage peptide library was passed through the tail vein of the mouse.
  • Peptide expression phage that specifically binds was selected.
  • a screening plan in this regard is shown in FIG. 2.
  • Figure 2 is a plan of M13 phage screening for peptide screening targeting cancer tissue, (1) to build a mouse model with cancer tissue formed on the right hind limbs, (2) 7 amino acids on the surface of M13 phage After circulating the phage library in which the constructed loop peptide library was expressed and circulated through the tail vein, (3) it was washed under various conditions, and (4) the targeted phage was finally eluted to obtain phage.
  • the phage obtained as described above was infected with E. coli, amplified, injected again through the mouse tail vein, circulated, and then repeated at a higher intensity washing condition, thereby repeating the process of finding a phage having a more specific and stronger binding force. (This is called biopanning).
  • Peptide Sequence Irradiation group (10 Gy) P1 TVRTSAD (SEQ ID NO: 1) P2 RYVGTLF (SEQ ID NO: 2) P3 NRGDRIL (SEQ ID NO: 3) Unirradiated controls W1 NWGDRIL (SEQ ID NO: 4) W2 QRSLPSL (SEQ ID NO: 5) W3 DVWHSAY (SEQ ID NO: 6)
  • the phages in which each loop-type peptide was obtained were subjected to amplification, and then, in order to confirm specific binding and target efficiency of peptide phage through in vivo imaging, a fluorescent probe was first attached. Specifically, 1 ⁇ g / ⁇ l of N-hydroxysuccinimide esters of Cy5.5 (Amersham) was added to 10 11 plaque forming units (pfu) in 1 mL of bicarbonate buffer (pH 8.3) and maintained in a dark environment for 3 hours. Phage surface proteins and Cy5.5 fluorescent probes were linked at room temperature.
  • Phage expressing each looped peptide linked to the Cy5.5 fluorescent probe was obtained by precipitating 170 ⁇ l of 20% (w / v) PEG 8000 / 2.5 M NaCl solution. To determine the rate at which Cy5.5 fluorescent probes were connected to each of the resulting phage samples, measured by the IVIS Spectrum Imaging System (Xenogen), and using the instrument's software program to determine the ROI values, It was confirmed that the ratio of Cy5.5 connections between phage samples was constant. The results are shown in FIG.
  • Each prepared loop-expressing phage was injected into the irradiated reactive xenograft mouse model and the control group constructed in Example 1 through the tail vein, respectively, and immediately after the injection, the image was measured up to 2 days in vivo. In the circulating pathway of the peptide and in other organs and tissues, the image was gradually removed and only targeted to cancer tissue was confirmed, thereby demonstrating complete targeting to gastric cancer tissue in vivo. 5 shows that the peptide sequence identified through the biopanning process in Example 2 exhibits excellent targeting ability in imaging results in vivo .
  • Example 4> selectively combined during irradiation Peptide To identify sequences in vivo Imaging
  • the target efficiency according to the irradiation was confirmed to prove that the sequence is a sequence reacting according to the cancer microenvironment upon irradiation.
  • the difference in irradiation was intended to determine whether the peptide was targeted according to the difference in the cancer microenvironment.
  • tumor-forming mice were transplanted by transplanting patient gastric cancer tissues, and only a part of them was irradiated to establish a control group and an experimental group.
  • Peptide expression phage selected in the same manner as in Example 3 was amplified and fluorescent labeling, and the same sample was injected into the irradiated mouse and its control mice to confirm the image on the last 2 days.
  • FIG. 7 shows that there is a difference in targeting efficiency when targeting in a mouse model irradiated with radiation, compared with targeting in a control mouse model, and the image measurement result of the embodiment is shown in FIG. 7.
  • the target efficiency was lowered in the mouse gastric tissue transplant mouse model used as a control, it was confirmed that the specific binding in the mouse model irradiated with the same patient gastric cancer tissue transplanted mouse.
  • each population was injected through the tail vein, and after 24 hours, cancer tissue was extracted and paraffin blocks and sections were prepared. Specifically, (1) the extracted cancer tissues were immersed in paraformaldehyde solution to fix the tissues at room temperature for 24 hours, then dehydrated and infiltrated with paraffin to form a paraffin block of cancer tissues, followed by 3 ⁇ m thickness using a microtome. Sectional sections were prepared by sectioning slides. In order to perform full-scale immunohistostaining, (2) deparaffinization was performed on the slice slide, and (3) the structure of various proteins immobilized in the tissue slice was restored so that the antibody-binding site could be restored normally. Unmasking process was performed.
  • DPPC dipalmitoylphosphatidylcholine, concentration 50 mM
  • DPPG-Na dipalmitoylphosphatidylglycerol, concentration 50 mM
  • lipids constituting liposomes DPPE-PDP (N- [3- (2Pyridinyldithio) -1-oxopropyl] -L- ⁇ -dipalmitoylphosphatidylcholine , 50mM)
  • cholesterol concentration 200mM
  • cholesterol-PEG concentration 200mM
  • DPPG-Na did not melt at room temperature, so it was completely dissolved at 55 ° C. for at least 30 minutes.
  • Each of the five dissolved lipids were mixed in a round bottom flask with DPPC: DPPE-PDP: DPPG-Na: Cholesterol-PEG: Cholesterol at a molar concentration of 15: 15: 30: 4: 36 and at 55 ° C.
  • HEPES (10 nM, pH4) buffer was added to prepare liposomes, the flask was rotated in a thermostat (55 ° C), dissolved for 1 hour, and shaken vigorously with a vortex to completely dissolve.
  • the 200 nm and 100 nm filters were extruded twice or several times.
  • the liposome was dissolved in PBS (pH 7.0) using a Sephadex column.
  • the inside of the liposome completed up to this process was pH 4.0, and the outside was pH 7.0.
  • After dissolving in the buffer mix the liposomes with the buffer substitution and the doxorubicin drug dissolved in the pH 7.0 buffer in a round flask and insert the dissolved drug in pH 7.0 into the liposome at pH 4.0.
  • the drug delivery system was optimized by measuring the size and stability of the final drug-encapsulated liposomes using the DLS method. As described above, the drug delivery process and the optimization process and the results are shown in FIG. 9.
  • Example 5 a peptide having a 'TVRTSAD' sequence was synthesized and subjected to a linking process with a liposome encapsulated with the drug in Example 6.
  • Cy7 fluorescent probe is connected to the C-terminus of the 'TVRTSAD' peptide, and the N-terminus was prepared by synthesizing a peptide which has not undergone any process in order to connect to liposomes.
  • the N-terminal residue of the prepared peptide was processed to be connected to a disulfide bond when the liposome was linked with a thiol group (-SH).
  • a liposome reduction experiment was conducted to identify the number of thiol groups in liposomes and to link them with peptides according to their ratios.
  • In vivo imaging was performed to determine whether the drug transporter linked to the drug-encapsulated liposome with the 'TVRTSAD' sequence processed in Example 6 actually works in vivo .
  • the injection-reactive xenograft mouse model and the control group constructed in Example 1 were injected through the tail vein, respectively, and immediately after the injection, the image was measured until the last 2 days, thereby circulating the pathways of peptides and other organs in vivo. And the image was gradually removed from the tissue and confirmed only target the cancer tissue, thereby demonstrating the complete target to the gastric cancer tissue in vivo.
  • In vivo imaging results performed according to the above embodiment are shown in FIG. 11.
  • in vivo tumor growth delay was performed for validation as a target drug carrier as well as in vivo imaging.
  • a new concept anticancer agent capable of diagnosis and treatment at the same time by verifying that the material made by linking the peptide with the drug-encapsulated liposome can be applied to in vivo imaging and in vivo tumor growth delay.
  • the results demonstrate the potential for development.
  • the discovery peptide selectively binds when irradiated in other patient gastric cancer tissues having the same characteristics as the same gastric cancer in addition to the gastric cancer case.
  • a mouse model was constructed by securing gastric cancer tissues extracted from other patients. Specifically, two kinds of gastric cancer tissues of other patients were obtained in addition to the patient gastric cancer tissues used in the above example, and it was confirmed that all the gastric cancer tissues used were carcinomas having the characteristics of adenocarcinoma. As in Example 1, a mouse model was constructed for each of the gastric cancer tissues, and some of them were irradiated to establish a control group and an experimental group.
  • Target efficiency according to the irradiation was verified.
  • Selection peptide expression phage amplification and fluorescence labeling was carried out in the same manner as in Example 3, and the final sample was injected into irradiated experimental mice and control mice, respectively, to confirm imaging on the last 2 days.
  • peptide phage was selectively accumulated only in tumors of the experimental group mice irradiated with radiation as in the other cases of gastric cancer tissues of the other patients, and the image measurement results of the example are shown in FIG. 13. .
  • FIG. 13 it was confirmed that the peptide expression phage does not target cancer tissue when not irradiated with radiation, but selectively targets cancer tissue when irradiated with radiation.
  • the peptide sequence selected in this technique was clearly verified as a peptide sequence that selectively targets gastric cancer tissues upon irradiation, and also the radiation activity of gastric cancer tissues of other patients was not investigated. Only if the peptide sequence exhibits a target ability selectively, it proved that the scope of application in clinical applications is not limited.

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Abstract

La présente invention concerne des peptides pour le ciblage du cancer gastrique, une composition pour le diagnostic du cancer gastrique basé sur une irradiation ou non-irradiation au moyen desdits peptides, et l'utilisation de ceux-ci pour l'administration d'un médicament. Les peptides fonctionnels capables de cibler le cancer ont été découverts en vue de la mise en œuvre de diagnostic et de traitement personnalisé pour des patients individuels en tenant compte de problèmes pendant le traitement dans quels cas des traitement de patients respectifs diffèrent en raison de différentes réponses thérapeutiques résultant de différences génétiques chez des patients individuels atteints de cancer. Après l'établissement de modèles animaux analogues aux micro-environnements de cancer de patients réels et la division en une population irradiée et une population non-irradiée en tant qu'un groupe témoin, l'efficacité de ciblage a été testée pour des peptides respectifs sélectionnés par le criblage de peptides de liaison spécifique aux populations respectives. En tant que telle, la présente invention peut enfin être utilisée dans le développement technique d'imagerie diagnostique pour la prédiction de la sensibilité à la radiothérapie et, par conséquent, le développement d'agents thérapeutiques ciblés personnalisés.
PCT/KR2015/007943 2014-11-18 2015-07-29 Peptides pour le ciblage de cancer gastrique, et leur utilisation médicale WO2016080633A1 (fr)

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US16/149,142 US10627402B2 (en) 2014-11-18 2018-10-02 Peptides for targeting gastric cancer, and medical use tehreof

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CN106699848A (zh) * 2017-03-10 2017-05-24 贵州医科大学 与激活型肝星状细胞特异性结合的多肽及制备方法和应用
IT201600090052A1 (it) * 2016-09-06 2018-03-06 Univ Degli Studi Padova Peptide per la diagnosi differenziale della Malattia di Crohn
US10124073B2 (en) 2014-08-04 2018-11-13 Case Western Reserve University Targeting peptides and methods of use
US10925980B2 (en) 2014-08-04 2021-02-23 Case Western Reserve University Molecular probes and methods of use

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080305212A1 (en) * 2007-04-16 2008-12-11 Solae, Llc Protein Hydrolysate Compositions Having Improved Sensory Characteristics and Physical Properties
CN102329374A (zh) * 2011-09-08 2012-01-25 西安交通大学 一种胃癌靶向性多肽及其应用
KR20140123311A (ko) * 2013-04-12 2014-10-22 울산대학교 산학협력단 암 표적용 펩타이드 및 이의 의학적 용도

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20080305212A1 (en) * 2007-04-16 2008-12-11 Solae, Llc Protein Hydrolysate Compositions Having Improved Sensory Characteristics and Physical Properties
CN102329374A (zh) * 2011-09-08 2012-01-25 西安交通大学 一种胃癌靶向性多肽及其应用
KR20140123311A (ko) * 2013-04-12 2014-10-22 울산대학교 산학협력단 암 표적용 펩타이드 및 이의 의학적 용도

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PASTOR, C. ET AL.: "Identification of major allergens in watermelon", INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY, vol. 149, 2009, pages 291 - 298, XP009503052 *
ZHANG, W. -J. ET AL.: "Affinity peptide developed by phage display selection for targeting gastric cancer", WORLD JOURNAL OF GASTROENTEROLOGY, vol. 18, no. 17, 2012, pages 2053 - 2060, XP055443272 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10124073B2 (en) 2014-08-04 2018-11-13 Case Western Reserve University Targeting peptides and methods of use
US10653801B2 (en) 2014-08-04 2020-05-19 Case Western Reserve University Targeting peptides and methods of use
US10925980B2 (en) 2014-08-04 2021-02-23 Case Western Reserve University Molecular probes and methods of use
US11738099B2 (en) 2014-08-04 2023-08-29 Case Western Reserve University Molecular probes and methods of use
IT201600090052A1 (it) * 2016-09-06 2018-03-06 Univ Degli Studi Padova Peptide per la diagnosi differenziale della Malattia di Crohn
WO2018047069A1 (fr) * 2016-09-06 2018-03-15 Universita' Degli Studi Di Padova Peptide pour le diagnostic différentiel de la maladie de crohn
CN106699848A (zh) * 2017-03-10 2017-05-24 贵州医科大学 与激活型肝星状细胞特异性结合的多肽及制备方法和应用
CN106699848B (zh) * 2017-03-10 2020-08-21 贵州医科大学 与激活型肝星状细胞特异性结合的多肽及制备方法和应用

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