CN116891828A - Novel immunostimulating molecules, modified exosomes, and preparation methods and applications thereof - Google Patents
Novel immunostimulating molecules, modified exosomes, and preparation methods and applications thereof Download PDFInfo
- Publication number
- CN116891828A CN116891828A CN202310364327.6A CN202310364327A CN116891828A CN 116891828 A CN116891828 A CN 116891828A CN 202310364327 A CN202310364327 A CN 202310364327A CN 116891828 A CN116891828 A CN 116891828A
- Authority
- CN
- China
- Prior art keywords
- csk
- pam
- exosome
- optionally
- exosomes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000001808 exosome Anatomy 0.000 title claims abstract description 139
- 230000003308 immunostimulating effect Effects 0.000 title claims abstract description 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 56
- 229960005486 vaccine Drugs 0.000 claims abstract description 26
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 14
- 238000011282 treatment Methods 0.000 claims abstract description 8
- 239000000427 antigen Substances 0.000 claims description 35
- 102000036639 antigens Human genes 0.000 claims description 35
- 108091007433 antigens Proteins 0.000 claims description 35
- 239000003814 drug Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 16
- 208000026278 immune system disease Diseases 0.000 claims description 10
- 201000010099 disease Diseases 0.000 claims description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 9
- 229940079593 drug Drugs 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 229910052760 oxygen Inorganic materials 0.000 claims description 9
- 229910052717 sulfur Inorganic materials 0.000 claims description 9
- 229960001438 immunostimulant agent Drugs 0.000 claims description 8
- 239000003022 immunostimulating agent Substances 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 7
- 108091008036 Immune checkpoint proteins Proteins 0.000 claims description 6
- 102000037982 Immune checkpoint proteins Human genes 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- IPCSVZSSVZVIGE-UHFFFAOYSA-N palmitic acid group Chemical group C(CCCCCCCCCCCCCCC)(=O)O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 238000002560 therapeutic procedure Methods 0.000 claims description 6
- 238000011534 incubation Methods 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 201000001441 melanoma Diseases 0.000 claims description 4
- 208000023275 Autoimmune disease Diseases 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 3
- 208000026310 Breast neoplasm Diseases 0.000 claims description 3
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 3
- 108091007741 Chimeric antigen receptor T cells Proteins 0.000 claims description 3
- 206010009944 Colon cancer Diseases 0.000 claims description 3
- 208000001333 Colorectal Neoplasms Diseases 0.000 claims description 3
- 208000000461 Esophageal Neoplasms Diseases 0.000 claims description 3
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 3
- 206010025323 Lymphomas Diseases 0.000 claims description 3
- 208000001894 Nasopharyngeal Neoplasms Diseases 0.000 claims description 3
- 206010061306 Nasopharyngeal cancer Diseases 0.000 claims description 3
- 206010030155 Oesophageal carcinoma Diseases 0.000 claims description 3
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 3
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 3
- 241000700605 Viruses Species 0.000 claims description 3
- 230000000903 blocking effect Effects 0.000 claims description 3
- 125000004432 carbon atom Chemical group C* 0.000 claims description 3
- 201000010881 cervical cancer Diseases 0.000 claims description 3
- 238000002512 chemotherapy Methods 0.000 claims description 3
- 201000004101 esophageal cancer Diseases 0.000 claims description 3
- 206010017758 gastric cancer Diseases 0.000 claims description 3
- 208000032839 leukemia Diseases 0.000 claims description 3
- 201000007270 liver cancer Diseases 0.000 claims description 3
- 208000014018 liver neoplasm Diseases 0.000 claims description 3
- 201000005202 lung cancer Diseases 0.000 claims description 3
- 208000020816 lung neoplasm Diseases 0.000 claims description 3
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 3
- 201000002528 pancreatic cancer Diseases 0.000 claims description 3
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 3
- 238000001959 radiotherapy Methods 0.000 claims description 3
- 201000011549 stomach cancer Diseases 0.000 claims description 3
- 206010044412 transitional cell carcinoma Diseases 0.000 claims description 3
- 230000000840 anti-viral effect Effects 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- 230000006806 disease prevention Effects 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 102000039446 nucleic acids Human genes 0.000 claims description 2
- 108020004707 nucleic acids Proteins 0.000 claims description 2
- 150000007523 nucleic acids Chemical class 0.000 claims description 2
- 229920001184 polypeptide Polymers 0.000 claims description 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 2
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 2
- 150000003384 small molecules Chemical class 0.000 claims description 2
- 239000012646 vaccine adjuvant Substances 0.000 claims description 2
- 229940124931 vaccine adjuvant Drugs 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 5
- 230000005909 tumor killing Effects 0.000 abstract description 5
- 230000004936 stimulating effect Effects 0.000 abstract description 3
- 230000001976 improved effect Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 16
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 12
- 239000002953 phosphate buffered saline Substances 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 10
- 239000002671 adjuvant Substances 0.000 description 10
- 210000004881 tumor cell Anatomy 0.000 description 9
- 230000009286 beneficial effect Effects 0.000 description 6
- 230000028993 immune response Effects 0.000 description 6
- 101150054980 Rhob gene Proteins 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 230000002708 enhancing effect Effects 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 102000014150 Interferons Human genes 0.000 description 3
- 108010050904 Interferons Proteins 0.000 description 3
- 102100027010 Toll-like receptor 1 Human genes 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000002648 combination therapy Methods 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 229940079322 interferon Drugs 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 230000004614 tumor growth Effects 0.000 description 3
- 101150013553 CD40 gene Proteins 0.000 description 2
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 2
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 2
- 108010060889 Toll-like receptor 1 Proteins 0.000 description 2
- 102000008228 Toll-like receptor 2 Human genes 0.000 description 2
- 108010060888 Toll-like receptor 2 Proteins 0.000 description 2
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 101000763579 Homo sapiens Toll-like receptor 1 Proteins 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 238000011398 antitumor immunotherapy Methods 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000011284 combination treatment Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 210000003162 effector t lymphocyte Anatomy 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000001024 immunotherapeutic effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000003071 memory t lymphocyte Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- DAZSWUUAFHBCGE-KRWDZBQOSA-N n-[(2s)-3-methyl-1-oxo-1-pyrrolidin-1-ylbutan-2-yl]-3-phenylpropanamide Chemical compound N([C@@H](C(C)C)C(=O)N1CCCC1)C(=O)CCC1=CC=CC=C1 DAZSWUUAFHBCGE-KRWDZBQOSA-N 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 238000003921 particle size analysis Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- HIGSLXSBYYMVKI-UHFFFAOYSA-N pralidoxime chloride Chemical compound [Cl-].C[N+]1=CC=CC=C1\C=N\O HIGSLXSBYYMVKI-UHFFFAOYSA-N 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000001235 sensitizing effect Effects 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000012876 topography Methods 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
- 210000003954 umbilical cord Anatomy 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0693—Tumour cells; Cancer cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- 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
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
-
- 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
- A61K47/51—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 the non-active ingredient being a modifying agent
- A61K47/62—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 the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
- C12N5/0626—Melanocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55516—Proteins; Peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55588—Adjuvants of undefined constitution
Abstract
The invention provides a novel immunostimulatory molecule, an improved exosome, a preparation method and application thereof. The provided immunostimulatory molecules include fatty chains and derivatives thereof, and the fatty chains and derivatives thereof are loaded on exosomes to obtain engineered exosomes. The immune stimulating molecule is combined with exosomes through a fatty chain, and after the exosomes from tumor sources are loaded with the fatty chain, an anti-tumor exosome vaccine can be prepared, so that tumor killing is promoted, and the immune treatment effect of tumors is enhanced.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to the field of exosome delivery immunostimulant and antitumor vaccine, and especially relates to a novel immunostimulant molecule, an exosome modified by the immunostimulant molecule, and a preparation method and application thereof.
Background
The interferon stimulating gene (STING) is an important cellular receptor and is expressed in the endoplasmic reticulum of a variety of immune cells and other cells. After STING is combined with the stimulator, the expression of immune molecules such as I-type interferon can be induced, and the immune response of the organism is started. Cyclic Dinucleotides (CDNs) are natural agonists of STING, which can interact with STING and cause activation of immune cells. The activated immune cells can generate I-type interferon and the like, can promote the organism to rapidly remove pathogens, and is also beneficial to presenting antigen to APCs and sensitizing effector T cells, thereby killing tumor cells. Thus, STING stimulators have an important role in anti-tumor therapy. However, natural CDNs have poor stability and low ability to cross cell membranes, and thus their use is limited.
Further improvements are needed based on STING pathway and its stimulators to enhance its use in the anti-tumor field.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the related art to some extent. According to the examples, pam was synthesized 3 CSK 4 -CDG SF And Pam is taken 3 CSK 4 -CDG SF The anti-tumor exosome vaccine is obtained by high-efficiently loading the anti-tumor exosome vaccine. The provided anti-tumor exosome vaccine can be used for preventing or treating diseases.
We have developedCombined adjuvant Pam based on STING pathway and Toll-like receptor 1/2 pathway 3 CSK 4 -CDG SF The combined adjuvant can remarkably improve the stability and transmembrane efficiency of CDNs. Pam 3 CSK 4 -CDG SF Has three palmitic acid chains, can bind to exosomes, and can be used for delivering Pam 3 CSK 4 -CDG SF Immunostimulant. The tumor-derived exosomes express antigen information secreted by tumor cells, and direct use of exosomes can elicit an antigen-specific immune response in the body. Tumor-derived exosome load Pam 3 CSK 4 -CDG SF After the adjuvant, an anti-tumor exosome vaccine can be prepared, and the exosome vaccine can simultaneously deliver antigen and the adjuvant to cause more effective immune response, thereby promoting tumor killing and being beneficial to enhancing the immune treatment effect of tumors. Exosomes loaded Pam without tumor antigen 3 CSK 4 -CDG SF The adjuvant can enhance nonspecific immunity activation, regulate organism immunity microenvironment, and is beneficial to tumor immunity killing.
Specifically, the invention provides the following technical scheme:
in a first aspect the invention provides an engineered exosome comprising a fatty chain and derivatives thereof, the fatty chain and derivatives thereof being loaded onto the exosome.
According to an embodiment of the present invention, the modified exosome provided above may further include the following technical features:
the number of the fatty chains may be 1,2 or 3, and the manner of connection between the plurality of fatty chains is not limited. According to embodiments of the invention, the fatty chain is 1-18 carbon atoms in length, for example 1,2,3,4,5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms.
According to an embodiment of the invention, the fatty chain and its derivatives are Pam 3 CSK 4 And derivatives thereof or other compounds that link drug small molecules, polypeptides, proteins, nucleic acids to fatty chains in any manner.
According to the inventionIn the embodiment of (2), the Pam 3 CSK 4 Derivatives include Pam 3 CSK 4 -RhoB、Pam 3 CSK 4 At least one of the CDNs or positional isomers thereof,
the Pam 3 CSK 4 The CDN or its positional isomer is:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any one of natural base A, T, C, G, U and non-natural base;
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH and-F.
According to an embodiment of the invention, the Pam 3 CSK 4 CDN is Pam 3 CSK 4 -CDG SF 。
According to an embodiment of the invention, the Pam 3 CSK 4 -CDG SF At least one of the three palmitic acid chains of (2) is loaded on the exosome.
According to an embodiment of the invention, the exosomes contain tumor antigens or no antigens, e.g. the exosomes contain tumor-associated antigens or tumor-specific antigens. The exosomes free of tumor antigens may be derived from normal tissue or body fluid cells.
According to an embodiment of the present invention, the tumor antigen is selected from any one of a blood tumor or a solid tumor cell including a tumor such as lung cancer, liver cancer, stomach cancer, esophageal cancer, colorectal cancer, cervical cancer, breast cancer, nasopharyngeal cancer, pancreatic cancer, melanoma, urothelial cancer, leukemia, and lymphoma.
The exosomes mentioned herein may be tumor cells or cell lines from a patient. The exosomes without tumor antigens are derived from normal tissues or body fluid cells, including exosomes derived from stem cells isolated from umbilical cord blood, umbilical cord, placenta, bone marrow, fat, blood, etc., and exosomes derived from blood, saliva, urine, ascites, etc., or exosomes derived from corresponding cell lines.
According to an embodiment of the invention, the Pam 3 CSK 4 -CDG SF At least one of the three palmitic acid chains is loaded on the exosomes.
According to an embodiment of the invention, the particle size of the modified exosomes ranges from 120 nm to 250 nm, preferably from 130 nm to 240 nm.
In a second aspect the invention provides a method of preparing the engineered exosome of the first aspect comprising:
the solution containing the exosomes and the solution containing the fatty chains and their derivatives are mixed and incubated in order to obtain the engineered exosomes.
According to an embodiment of the present invention, the preparation method of the modified exosome described above may further include the following technical features:
according to an embodiment of the invention, the incubation temperature is 4 degrees celsius; preferably, the incubation time is 2 to 5 hours.
According to an embodiment of the invention, each of the solution containing exosomes or the solution containing fatty chains and derivatives thereof is independently a PBS solution.
In a third aspect, the present invention provides an immunostimulatory molecule having the chemical formula:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any one of natural base A, T, C, G, U and non-natural base;
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH and-F.
In a fourth aspect, the invention provides the use of an immunostimulatory molecule of the third aspect in the manufacture of a medicament for antiviral, antitumor therapy and vaccine adjuvants.
In a fifth aspect, the invention provides an engineered exosome comprising Pam 3 CSK 4 And derivatives thereof, the Pam 3 CSK 4 And derivatives thereof are loaded on exosomes.
The Pam 3 CSK 4 Derivatives include Pam 3 CSK 4 -RhoB、Pam 3 CSK 4 At least one of the CDNs or positional isomers thereof,
the Pam 3 CSK 4 The CDN or its positional isomer is:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any one of natural base A, T, C, G, U and non-natural base;
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH and-F.
According to an embodiment of the invention, the Pam 3 CSK 4 CDN is Pam 3 CSK 4 -CDG SF 。
A sixth aspect of the invention provides an exosome vaccine comprising the engineered exosome of the first aspect or comprising the engineered exosome obtained according to the method of preparation of any of the second aspect or comprising the engineered exosome of the fifth aspect.
A seventh aspect of the invention provides the use of an altered exosome according to the first aspect of the invention or obtained according to the method of preparation according to the second aspect of the invention or comprising an altered exosome according to the fifth aspect in the field of disease prevention and/or treatment.
An eighth aspect of the present invention provides a pharmaceutical combination composition comprising: the modified exosome of the first or fifth aspect; and at least one drug for preventing and/or treating immune diseases. The modified exosomes and the medicament for preventing and/or treating immune diseases may be formulated into a composition for administration together to a subject in need thereof for the treatment of diseases such as tumors, viruses, etc. The modified exosomes and the medicament for preventing and/or treating immune diseases may be administered together or sequentially, for example, the modified exosomes may be administered first followed by the medicament for preventing and/or treating immune diseases or the medicament for preventing and/or treating immune diseases may be administered first followed by the modified exosomes.
According to a specific embodiment, the at least one drug for preventing and/or treating an immune disease is selected from at least one of immune checkpoint blocking antibodies, immunostimulants, vaccines, chimeric antigen receptor T cells, radiation therapy and chemotherapy.
In a ninth aspect the present invention provides a method of preventing and/or treating a disease comprising administering to a subject in need thereof an effective amount of a medicament which is an altered exosome according to the first or sixth aspect of the invention or a pharmaceutical combination composition according to the eighth aspect.
According to a specific embodiment, the disease is at least one of a tumor, a virus or an autoimmune disease.
The beneficial effects obtained by the invention are as follows:
1) The antigen carried by exosomes is used, so that identification, analysis and preparation of tumor antigens are not needed, and the preparation process of the anti-tumor vaccine is greatly simplified;
2) With Pam 3 CSK 4 -CDG SF For example, pam on exosomes 3 CSK 4 -CDG SF The loading efficiency of the vaccine is up to more than 99%, so that the preparation process of the exosome vaccine is greatly simplified, and the utilization rate of the adjuvant molecules is also improved;
3) The exosome vaccine can simultaneously deliver Pam 3 CSK 4 Or derivatives thereof (e.g. Pam 3 CSK 4 -CDG SF Adjuvant) and antigen, enhancing the immune activating ability, and improving the immune therapeutic effect.
Drawings
FIG. 1 is a Pam-based provided in accordance with an embodiment of the present invention 3 CSK 4 -CDG SF Is a schematic of an exosome vaccine.
FIG. 2 is a Pam provided in accordance with an embodiment of the present invention 3 CSK 4 -CDG SF Particle size analysis of (3).
FIG. 3 is a Pam provided in accordance with an embodiment of the present invention 3 CSK 4 -CDG SF Is a topography analysis chart of (3).
Fig. 4 is a graph of standard curve results provided in accordance with an embodiment of the present invention.
FIG. 5 is a Pam provided in accordance with an embodiment of the present invention 3 CSK 4 -CDG SF Pam 3 CSK 4 -CDG SF The @ EXO stimulated RAW 264.7 cells to express CD40 (A) and CD80 (B).
Fig. 6 is a graph of statistics of cd4+ and cd8+cd44+cd62l in spleen provided according to an embodiment of the present invention.
Fig. 7 is a graph showing the statistical result of survival rate of tumor-bearing mice according to an embodiment of the present invention.
Fig. 8 is a graph of survival statistics of tumor-bearing mice treated with combination therapy provided according to an embodiment of the present invention.
Fig. 9 is a graph of tumor growth curve statistics of tumor-bearing mice treated by combination therapy provided according to an embodiment of the present invention.
Detailed Description
The following detailed description of embodiments of the invention, examples of which are illustrated in the accompanying drawings, it being noted that the embodiments described with reference to the drawings are illustrative and intended to be used for explaining the invention and are not to be construed as limiting the invention.
Herein, the tumor antigens mentioned include tumor-associated antigens and tumor-specific antigens.
Tumor Associated Antigen (TAA) refers to a protein, glycoprotein or lipoprotein that is immunogenic on the surface of tumor cells. The tumor antigen can be used as a tumor marker for detecting and diagnosing tumors, and can also be used for distinguishing tumor-derived exosomes or normal cell-derived exosomes.
Tumor specific antigens (tumor specific antigen, TSA) are antigens that are all of the tumor cells but not present in normal cells, with higher tumor specificity.
The invention provides an immunostimulatory molecule, which has a chemical structural formula as follows:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any of natural base A, T, C, G, U and non-natural base:
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH and-F.
The provided immunostimulatory molecule can be used as a combined adjuvant of STING channel and Toll-like receptor 1/2 channel, can play a role in immunostimulatory, can remarkably activate APC cells and activate immune response, and can be applied to immunotherapy. According to a specific embodiment, n is 1,2,3,4,5,6 or 7. According to a specific embodiment, Y 1 Is O, Y 2 Is O; or Y 1 Is O, Y 2 S is the same as the original formula; or Y 1 Is S, Y 2 Is O; or Y 1 Is S, Y 2 S. According to a specific embodiment, X 1 Is H, X 2 Is H, or X 1 is-OH, X 2 is-H, or X 1 is-F, X 2 is-H, or X 1 is-OH, X 2 Is F, or X 1 is-F, X 2 is-F. The immune stimulating molecule is combined with exosomes through a fatty chain, and after the exosomes from tumor sources are loaded with the fatty chain, an anti-tumor exosome vaccine can be prepared, so that tumor killing is promoted, and the immune treatment effect of tumors is enhanced.
The invention provides a modified exosome comprising: pam 3 CSK 4 -CDN or its positional isomer, said Pam 3 CSK 4 The CDN or its positional isomer is loaded on the exosomes. Pam mentioned 3 CSK 4 The chemical structural formula of the CDN or its positional isomer is shown above.
The invention provides a modified exosome comprising: pam 3 CSK 4 -CDG SF The Pam 3 CSK 4 -CDG SF Loaded on the exosome. The exosomes are tumor-derived exosomes, antigen information secreted by tumor cells is expressed on the tumor-derived exosomes, and direct use of the exosomes can causeAntigen-specific immune response of the body. Tumor-derived exosome load Pam 3 CSK 4 -CDG SF After the adjuvant, an anti-tumor exosome vaccine can be prepared, and the exosome vaccine can simultaneously deliver antigen and the adjuvant to cause more effective immune response, thereby promoting tumor killing and being beneficial to enhancing the immune treatment effect of tumors. According to a specific embodiment of the invention, the exosomes contain tumor antigens thereon. Such tumor antigens include, but are not limited to, B16-OVA. Exosomes free of tumor antigens may also be used as carriers for the delivery of immunostimulants, such as Pam 3 CSK 4 -CDG SF And derivatives thereof.
According to a specific embodiment of the invention, the Pam 3 CSK 4 -CDG SF At least one of the three palmitic acid chains is loaded on the exosomes. For example, one, two or three of the three palmitic acid chains are loaded on the exosomes. The loading mentioned may be on exosome lipid bilayer membranes.
Pam mentioned 3 CSK 4 -CDG SF The chemical structural formula is shown below, which can be synthesized according to a method commonly used in the art. For example, reference may be made to Hu H G, wu J J, zhang B D, li W H, li Y M.Pam 3 CSK 4 -CDG SF Augments Antitumor Immunotherapy by Synergistically Activating TLR1/2and STING[J]Pam is prepared by the method described in Bioconjugate Chemistry,2020,31,2499-2499 3 CSK 4 -CDG SF . Other Pam 3 CSK 4 The derivatives may also be synthesized by the methods described in the literature. Pam mentioned 3 CSK 4 Derivatives include Pam 3 CSK 4 、Pam 3 CSK 4 -RhoB、Pam 3 CSK 4 -CDG SF At least one of them.
Wherein Pam is 3 CSK 4- CDG SF Wherein n is 1,2,3,4,5,6 or 7.
According to a specific embodiment of the invention, the exosome vaccine has an average particle size of 120 nm to 250 nm, preferably 130 nm to 240 nm. For example, the engineered exosomes have an average particle size of about 120 nm, 130 nm, 140 nm, 150 nm, 160 nm, 170 nm, 180 nm, 190 nm, 200 nm, 210 nm, 220 nm, 230 nm, 240 nm, 250 nm, etc.
The invention also provides a method for preparing the modified exosome, which comprises the following steps: mixing exosome-containing solution with Pam-containing solution 3 CSK 4 -CDG SF To obtain an engineered exosome.
According to a specific embodiment of the invention, the incubation is performed at a low temperature, for example at 4 degrees celsius for 2 to 5 hours.
The solution containing exosomes is PBS solution containing exosomes, and the solution contains Pam 3 CSK 4 -CDG SF Is a solution containing Pam 3 CSK 4 -CDG SF Is a solution of PBS.
The modified exosome can be used as exosome vaccine, applied to patients, can promote tumor killing, and is beneficial to enhancing the immunotherapeutic effect of tumors. Can also be combined with other medicines for preventing and/or treating immune diseases to form pharmaceutical combination composition. To this end the present invention provides a pharmaceutical combination composition comprising: the modified exosomes; and at least one drug for preventing and/or treating a disease. According to a specific embodiment, the at least one drug for preventing and/or treating an immune disease is selected from at least one of immune checkpoint blocking antibodies, immunostimulants, vaccines, chimeric antigen receptor T cells, radiation therapy and chemotherapy. The exosomes or exosome vaccines or pharmaceutical combination compositions provided may be applied in a variety of disease fields including, but not limited to, tumors, autoimmune diseases. The tumor includes, but is not limited to, any one of blood tumor or solid tumor including lung cancer, liver cancer, stomach cancer, esophageal cancer, colorectal cancer, cervical cancer, breast cancer, nasopharyngeal cancer, pancreatic cancer, melanoma, urothelial cancer, leukemia, lymphoma, etc.
To this end, the present invention also provides a method for preventing and/or treating a disease, comprising: administering to a subject in need thereof a prophylactically and/or therapeutically effective amount of the exosome vaccine described above or the exosome or the pharmaceutical combination composition described above. The therapeutically effective amounts mentioned may be 3 to 8 nanomolar Pam of exosome load containing 10 to 40 micrograms of protein 3 CSK 4 -CDG SF . For example, an exosome containing 20. Mu.g of protein may be loaded with 5 nanomolar Pam 3 CSK 4 -CDG SF 。
It will be appreciated by those skilled in the art that the following examples are illustrative of the present invention and should not be construed as limiting the scope of the invention. The examples are not to be construed as limiting the specific techniques or conditions described in the literature in this field or as per the specifications of the product. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1
We synthesized Pam 3 CSK 4 -CDG SF Extracting exosomes derived from B16-OVA cells (mouse melanoma cells), and mixing and incubating to obtain Pam 3 CSK 4 -CDG SF Is efficiently loaded on exosomes to prepare antitumor exosome vaccine (Pam 3 CSK 4 -CDG SF @ EXO), as shown in fig. 1. The method specifically comprises the following steps:
diluting the extracted exosomes into a suitable PBS solution, adding a suitable amount of Pam 3 CSK 4 -CDG SF Incubating the solution in PBS for 3 hours at 4 ℃ to obtain Pam 3 CSK 4 -CDG SF @EXO。
Example 2
Example 2 Pam prepared for example 1 3 CSK 4 -CDG SF @ EXO is characterized as follows:
1. particle size and morphology
Particle size characterization was performed by Nanoparticle Tracking Analysis (NTA) method.
The results show that: pam 3 CSK 4 -CDG SF The particle size of @ EXO is 189.9.+ -. 50.2nm (see FIG. 2).
Morphology characterization was performed by Transmission Electron Microscopy (TEM).
The results show that: pam 3 CSK 4 -CDG SF The @ EXO shape is in the form of an irregular pie (as shown in fig. 3).
2. Load amount
To verify Pam 3 CSK 4 Interaction with exosomes, measurement of exosomes versus Pam 3 CSK 4 We synthesized Pam with rhodamine fluorophores 3 CSK 4 -RhoB。Pam 3 CSK 4 The PBS solution of RhoB was mixed with the exosome solution, incubated at 4℃for 3 hours, and the supernatant was centrifuged at 13000rpm for 10 minutes. According to Pam 3 CSK 4 Characteristic absorbance of RhoB at 488nm, standard curve was prepared and Pam was measured in the supernatant 3 CSK 4 -RhoB content, calculation of exosomes for Pam of different concentrations 3 CSK 4 The loading rate of RhoB is greater than 99%.
Pam 3 CSK 4 RhoB molecular Structure
The standard curve is shown in fig. 4.
B16-OVA exosome pair Pam 3 CSK 4 The loading results of RhoB are shown in Table 1 below.
TABLE 1
Example 3
Example 3 Pam prepared for example 1 3 CSK 4 -CDG SF The @ EXO performs functional verification.
1、Pam 3 CSK 4 -CDG SF EXO activates immune cells.
Using Pam 3 CSK 4 -CDG SF The @ EXO stimulated mouse macrophages, and the experimental groups were:
blank control group: PBS (phosphate buffered saline)
Exosome group: EXO (EXO)
Exosome mixing Pam 3 CSK 4 Group: pam 3 CSK 4 +EXO
Exosome load Pam 3 CSK 4 Group: pam 3 CSK 4 @EXO
Exosome mixing Pam 3 CSK 4 -CDG SF Group: pam 3 CSK 4- CDG SF +EXO
Exosome load Pam 3 CSK 4 -CDG SF Group: pam 3 CSK 4- CDG SF @EXO。
As a result, it was found that it can increase the expression of activated molecules such as CD40 and CD80 and enhance the secretion of cytokines (as shown in FIG. 5).
2、Pam 3 CSK 4 -CDG SF EXO activates memory T cells in mice:
using Pam 3 CSK 4 -CDG SF The @ EXO immunized C57/BL6 mice were three times. The experimental groups are respectively:
blank control group: PBS (phosphate buffered saline)
Tumor cell lysate antigen group: lysate
Exosome group: EXO (EXO)
Tumor cell lysate antigen mixed Pam 3 CSK 4 -CDG SF Group: pam 3 CSK 4- CDG SF +lysate
Exosome mixing Pam 3 CSK 4 -CDG SF Group: pam 3 CSK 4- CDG SF +EXO
Exosome load Pam 3 CSK 4 -CDG SF Group: pam 3 CSK 4- CDG SF @EXO。
It was found that memory T in its spleen was significantly activated (as shown in fig. 6).
3、Pam 3 CSK 4 -CDG SF @ EXO treatment of tumors.
Using Pam 3 CSK 4 -CDG SF The @ EXO immune tumor-bearing mice can effectively improve the survival rate of the tumor-bearing mice (shown in figure 7).
4、Pam 3 CSK 4 -CDG SF EXO is used in combination with immune checkpoint therapy to treat tumors.
Using Pam 3 CSK 4 -CDG SF Exosome vaccines can better inhibit tumor growth by combining with immune checkpoint therapy by immunizing tumor-bearing mice twice with EXO-PD 1 and prolonging mice survival (as shown in fig. 8 and 9).
Blank control group: PBS (phosphate buffered saline)
Immune checkpoint therapy group: anti-PD1
Exosome load Pam 3 CSK 4 -CDG SF Group: pam 3 CSK 4 -CDG SF @EXO
Combination therapy group Pam 3 CSK 4 -CDG SF @EXO+anti-PD1。
The experimental results show that: exosome load Pam 3 CSK 4 -CDG SF The mice had higher survival rates, smaller tumor volumes, and slower tumor growth than the anti-PD-1 and placebo groups. The combination treatment group showed more excellent effects.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.
While embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and not to be construed as limiting the invention, and that variations, modifications, alternatives and variations may be made to the above embodiments by one of ordinary skill in the art within the scope of the invention.
Claims (10)
1. An engineered exosome comprising a fatty chain and derivatives thereof,
the fatty chain and the derivative thereof are loaded on an exosome;
optionally, the fatty chain is 1 to 18 carbon atoms in length;
optionally, the fatty chain may be at least one, preferably 1,2 or 3.
2. The engineered exosome of claim 1, wherein the fatty chain and derivatives thereof are Pam 3 CSK 4 And derivatives thereof or other compounds that link drug small molecules, polypeptides, proteins, nucleic acids to fatty chains in any manner;
optionally, the Pam 3 CSK 4 Derivatives include Pam 3 CSK 4 -RhoB、Pam 3 CSK 4 At least one of the CDNs or positional isomers thereof,
the Pam 3 CSK 4 The CDN or its positional isomer is:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any one of natural base A, T, C, G, U and non-natural base;
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH, and-F;
optionally, the Pam 3 CSK 4 CDN is Pam 3 CSK 4 -CDG SF ；
Optionally, the Pam 3 CSK 4 -CDG SF At least one of the three palmitic acid chains of (2) is loaded on the exosome;
optionally, the exosomes contain or are free of tumor antigens;
optionally, the tumor antigen contained on the exosomes comprises a tumor-associated antigen and a tumor-specific antigen;
optionally, the tumor antigen is derived from any one selected from lung cancer, liver cancer, gastric cancer, esophageal cancer, colorectal cancer, cervical cancer, breast cancer, nasopharyngeal cancer, pancreatic cancer, melanoma, urothelial cancer, leukemia, and lymphoma;
optionally, the particle size of the engineered exosomes ranges from 120 nm to 250 nm, preferably 130 nm to 240 nm.
3. A method of preparing the engineered exosome of claims 1 or 2, comprising:
mixing and incubating a solution containing exosomes with a solution containing fatty chains and derivatives thereof in order to obtain engineered exosomes;
optionally, the incubation temperature is 4 degrees celsius; preferably, the incubation time is 2 to 5 hours;
optionally, each of the exosome-containing solution or the fatty chain and derivative-containing solution is independently a PBS solution.
4. An immunostimulatory molecule, wherein the immunostimulatory molecule has a chemical formula of:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any one of natural base A, T, C, G, U and non-natural base;
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH and-F.
5. Use of an immunostimulatory molecule according to claim 4 in the field of manufacture of a medicament, wherein the medicament is capable of use in antiviral, antitumor therapy and vaccine adjuvants.
6. An altered exosome comprising Pam 3 CSK 4 And the derivatives thereof, and the use thereof,
the Pam 3 CSK 4 And derivatives thereof are loaded on exosomes;
optionally, the Pam 3 CSK 4 Derivatives include Pam 3 CSK 4 -RhoB、Pam 3 CSK 4 At least one of the CDNs or positional isomers thereof,
the Pam 3 CSK 4 The CDN or its positional isomer is:
or a positional isomer thereof
Wherein B is 1 And B 2 Each independently selected from any one of natural base A, T, C, G, U and non-natural base;
n=any of 1 to 7;
Y 1 and Y 2 Each independently selected from any one of O and S;
X 1 and X 2 Each independently selected from any one of-H, -OH, and-F;
optionally, the Pam 3 CSK 4 CDN is Pam 3 CSK 4 -CDG SF 。
7. An exosome vaccine comprising the engineered exosome of claim 1 or 2 or comprising the engineered exosome obtained according to the method of preparation of claim 3 or comprising the engineered exosome of claim 6.
8. Use of an engineered exosome in the field of disease prevention and/or treatment, said engineered exosome being an engineered exosome according to claim 1 or 2 or an engineered exosome obtained according to the method of preparation according to claim 3 or an engineered exosome according to claim 6.
9. A pharmaceutical combination composition comprising:
the engineered exosome of claims 1 or 2 or 6; and at least one drug for preventing and/or treating immune diseases;
optionally, the at least one drug for preventing and/or treating an immune disease is selected from at least one of immune checkpoint blocking antibodies, immunostimulants, vaccines, chimeric antigen receptor T cells, radiation therapy and chemotherapy.
10. A method of preventing and/or treating a disease comprising administering to a subject in need thereof an effective amount of the modified exosome of claim 1 or 2 or 6, or the pharmaceutical combination composition of claim 9;
optionally, the disease is at least one of a tumor, a virus, or an autoimmune disease.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210357610 | 2022-04-06 | ||
| CN2022103576101 | 2022-04-06 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116891828A true CN116891828A (en) | 2023-10-17 |
Family
ID=88312620
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310364327.6A Pending CN116891828A (en) | 2022-04-06 | 2023-04-06 | Novel immunostimulating molecules, modified exosomes, and preparation methods and applications thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116891828A (en) |
-
2023
- 2023-04-06 CN CN202310364327.6A patent/CN116891828A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Li et al. | Nanovaccines integrating endogenous antigens and pathogenic adjuvants elicit potent antitumor immunity | |
| US20030138413A1 (en) | Methods for treating cancer | |
| JP2024016203A (en) | Antigenic peptides for cancer prevention and treatment | |
| US20230201322A1 (en) | Immunogenic compounds for cancer therapy | |
| US20230080443A1 (en) | Immunogenic compounds for cancer therapy | |
| JP2004538000A (en) | Methods for maturation of dendritic cells | |
| TWI719351B (en) | Nano-particles that contain synthetic variants of gm3 ganglioside as adjuvants in vaccines | |
| WO2016163489A1 (en) | Medicine | |
| KR20090047290A (en) | Exosome and composition of cancer vaccine containing it | |
| EP3536340A1 (en) | Autologous cancer cell vaccine | |
| KR20090047289A (en) | Exosome and composition of cancer vaccine containing it | |
| CN116891828A (en) | Novel immunostimulating molecules, modified exosomes, and preparation methods and applications thereof | |
| Hearnden et al. | Adjuvant strategies for vaccines: the use of adjuvants within the cancer vaccine setting | |
| CN114177282B (en) | Use of fluorinated polyethylenimine for preparing vaccine or preparation for preventing/treating diseases caused by virus/bacteria | |
| US20180055920A1 (en) | Vaccine, therapeutic composition and methods for treating or inhibiting cancer | |
| AU2006200116B2 (en) | Methods for treating cancer using a combination of a tumor-derived dendritic cell inhibitory factor antagonist and a toll-like receptor agonist |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |