CN116492462B - Application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation - Google Patents
Application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation Download PDFInfo
- Publication number
- CN116492462B CN116492462B CN202310088670.2A CN202310088670A CN116492462B CN 116492462 B CN116492462 B CN 116492462B CN 202310088670 A CN202310088670 A CN 202310088670A CN 116492462 B CN116492462 B CN 116492462B
- Authority
- CN
- China
- Prior art keywords
- cornea
- immune rejection
- transplantation
- bms
- cornea transplantation
- 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.)
- Active
Links
- 238000002054 transplantation Methods 0.000 title claims abstract description 35
- 210000004087 cornea Anatomy 0.000 title abstract description 60
- 229940122298 Peptidyl arginine deiminase IV inhibitor Drugs 0.000 title abstract description 12
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 17
- 239000003814 drug Substances 0.000 claims abstract description 16
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 238000000034 method Methods 0.000 claims description 6
- 239000007943 implant Substances 0.000 abstract description 22
- 230000015572 biosynthetic process Effects 0.000 abstract description 14
- 229940079593 drug Drugs 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 6
- 230000004083 survival effect Effects 0.000 abstract description 4
- 230000002035 prolonged effect Effects 0.000 abstract description 3
- 210000000795 conjunctiva Anatomy 0.000 abstract description 2
- 201000010099 disease Diseases 0.000 abstract description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 14
- 238000006243 chemical reaction Methods 0.000 description 12
- 239000007924 injection Substances 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 230000000735 allogeneic effect Effects 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 210000000440 neutrophil Anatomy 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 102100034540 Adenomatous polyposis coli protein Human genes 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 210000000265 leukocyte Anatomy 0.000 description 4
- 239000004033 plastic Substances 0.000 description 4
- 229920003023 plastic Polymers 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000011144 upstream manufacturing Methods 0.000 description 4
- 101100406797 Arabidopsis thaliana PAD4 gene Proteins 0.000 description 3
- 239000004475 Arginine Substances 0.000 description 3
- 238000011725 BALB/c mouse Methods 0.000 description 3
- 201000004569 Blindness Diseases 0.000 description 3
- 108010033040 Histones Proteins 0.000 description 3
- 102100037850 Interferon gamma Human genes 0.000 description 3
- 108010074328 Interferon-gamma Proteins 0.000 description 3
- 101150094373 Padi4 gene Proteins 0.000 description 3
- 102100035731 Protein-arginine deiminase type-4 Human genes 0.000 description 3
- 210000001744 T-lymphocyte Anatomy 0.000 description 3
- 210000002159 anterior chamber Anatomy 0.000 description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000003889 eye drop Substances 0.000 description 3
- 229940012356 eye drops Drugs 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 230000000144 pharmacologic effect Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000001356 surgical procedure Methods 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 102000006947 Histones Human genes 0.000 description 2
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 2
- 102000003777 Interleukin-1 beta Human genes 0.000 description 2
- 108090000193 Interleukin-1 beta Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 206010030113 Oedema Diseases 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 2
- 229940124599 anti-inflammatory drug Drugs 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 210000000612 antigen-presenting cell Anatomy 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 210000001508 eye Anatomy 0.000 description 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 2
- 230000001744 histochemical effect Effects 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 238000004020 luminiscence type Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000003801 milling Methods 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 229920002113 octoxynol Polymers 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 230000000149 penetrating effect Effects 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 238000003044 randomized block design Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- GSDSWSVVBLHKDQ-UHFFFAOYSA-N 9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij]quinoline-6-carboxylic acid Chemical compound FC1=CC(C(C(C(O)=O)=C2)=O)=C3N2C(C)COC3=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-UHFFFAOYSA-N 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- 239000012110 Alexa Fluor 594 Substances 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 208000002177 Cataract Diseases 0.000 description 1
- 206010011017 Corneal graft rejection Diseases 0.000 description 1
- 229930105110 Cyclosporin A Natural products 0.000 description 1
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 1
- 108010036949 Cyclosporine Proteins 0.000 description 1
- 241000283074 Equus asinus Species 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 206010022945 Iris adhesions Diseases 0.000 description 1
- 102000003896 Myeloperoxidases Human genes 0.000 description 1
- 108090000235 Myeloperoxidases Proteins 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 238000009004 PCR Kit Methods 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 102000016387 Pancreatic elastase Human genes 0.000 description 1
- 108010067372 Pancreatic elastase Proteins 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 239000006180 TBST buffer Substances 0.000 description 1
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 description 1
- 206010052779 Transplant rejections Diseases 0.000 description 1
- 208000024248 Vascular System injury Diseases 0.000 description 1
- 208000012339 Vascular injury Diseases 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000003409 anti-rejection Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 229960001265 ciclosporin Drugs 0.000 description 1
- 230000006329 citrullination Effects 0.000 description 1
- -1 compound topiramate Chemical class 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 208000030533 eye disease Diseases 0.000 description 1
- 239000003885 eye ointment Substances 0.000 description 1
- 210000000744 eyelid Anatomy 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 210000002443 helper t lymphocyte Anatomy 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 230000009773 lymphatic migration Effects 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 230000002911 mydriatic effect Effects 0.000 description 1
- 229960001699 ofloxacin Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 210000001747 pupil Anatomy 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000013077 scoring method Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229960001967 tacrolimus Drugs 0.000 description 1
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 229960004394 topiramate Drugs 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 208000008918 voyeurism Diseases 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0048—Eye, e.g. artificial tears
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- 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
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Transplantation (AREA)
- Ophthalmology & Optometry (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention provides application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation, belonging to the technical field of ophthalmic disease medicines. The invention provides an application of a preparation for inhibiting NETs formation in preparing medicines for delaying and/or inhibiting immune rejection after cornea transplantation, wherein PAD4 inhibitor is innovatively applied to preventing and treating the immune rejection after cornea transplantation, for example, BMS-P5 with a certain dosage is injected under conjunctiva, so that the immune rejection of cornea transplantation can be effectively inhibited, the survival time of cornea implants can be prolonged, the preparation can be used for treating the immune rejection after cornea transplantation, and good effects can be obtained.
Description
Technical Field
The invention belongs to the technical field of ophthalmic disease medicines, and particularly relates to application of a PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation.
Background
Cornea blindness is a second approximate blindness eye disease in China, at present, about 400 ten thousand of cornea blindness patients in China can be recovered through cornea transplantation operation, but immune rejection reaction after cornea transplantation operation is caused by transplantation lossThe most significant cause of failure. Immune rejection of cornea transplantations is mainly composed of CD4 + T cell mediated, its process mainly includes: antigen presenting cells (Antigenpresenting Cell, APC) recognize corneal antigens, transmit signals to cervical lymph nodes, proliferate T cells, activate aggressive helper T cells (Th), and then attack corneal implants via vascular or lymphatic migration to cause immune rejection. At present, medicaments such as glucocorticoid, cyclosporin A eye drops, tacrolimus eye drops and the like are mainly applied clinically to prevent and treat cornea transplantation immune rejection reaction, but the anti-rejection effect is not ideal. The main reason is that the mechanism of the antagonism rejection reaction of the medicine is mainly targeted to inhibit lymphocyte proliferation, namely, the mechanism plays a role in the middle link of the immune rejection reaction, but does not interfere with the source of the initial link of immune signal recognition. Therefore, how to perform effective intervention strategies in the initial stage of the initiation of the immune rejection reaction, thereby blocking the subsequent immune response, and having important clinical application value for preventing and treating the cornea transplantation immune rejection reaction and improving the prognosis of the cornea blind patient.
The initiation of immune rejection is initiated by antigen signaling and activation by APC followed by activation of primary CD4 either directly or indirectly + T cells. While it has been recognized that APC plays a key role in mediating graft immune rejection, the specific molecular mechanism of APC activation during immune rejection is not yet understood. Studies have shown that neutrophil extracellular traps (neutrophil extracellulartraps, nes) promote APC activation, initiate an immune response in the body, and thus cause tissue damage after organ transplantation. NETs are composed mainly of extracellular DNA and granular antimicrobial proteins, including citrullinated histones, elastase, myeloperoxidase, and the like. Protein arginine deaminase 4 (PAD 4) -driven histone citrullination is required in the process of forming the nes, and studies show that inhibiting PAD4 activity can block the formation of the nes, regulate innate immune responses and alleviate vascular injury in atherosclerosis mice. However, the effectiveness of PAD4 inhibitors for treating immune rejection following corneal transplantation has not been reported.
Disclosure of Invention
The invention aims to provide an application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation, and the PAD4 inhibitor is used for preparing a medicament for inhibiting the immune rejection after cornea transplantation, so that the effect is good.
The invention provides an application of a preparation for inhibiting the formation of Neutrophil Extracellular Traps (NETs) in preparing a medicament for delaying and/or inhibiting immune rejection after cornea transplantation.
Preferably, the agent that inhibits neutrophil extracellular trap formation comprises an arginine deaminase 4 (PAD 4) inhibitor.
Preferably, the agent that inhibits neutrophil extracellular trap formation comprises BMS-P5.
Preferably, the pharmacologically effective concentration of BMS-P5 is 4-60 mg/ml.
Preferably, the pharmacologically effective concentration of BMS-P5 is 5mg/mL.
The invention also provides application of BMS-P5 in preparing anti-inflammatory drugs for inhibiting I-type IFN related channels.
The invention also provides a medicament for inhibiting cornea transplant immune rejection reaction, which comprises BMS-P5 with pharmacological effective concentration.
Preferably, the medicament is in the form of injection.
The beneficial effects are that: the invention provides an application of a preparation for inhibiting NETs formation in preparing medicines for delaying and/or inhibiting immune rejection after cornea transplantation, wherein PAD4 inhibitor is innovatively applied to prevention and treatment of immune rejection after cornea transplantation, PAD4 inhibitor BMS-P5 is selected in examples to prevent and treat immune rejection after cornea transplantation, NETs expression is increased in cornea implants for generating immune rejection, and after allogeneic mice cornea transplantation, BMS-P5 (such as 5 mg/mL) is injected under conjunctiva at a certain dose, so that the immune rejection after cornea transplantation can be effectively inhibited, the survival time of the cornea implants can be prolonged, and the preparation can be used for treatment of immune rejection after cornea transplantation, and good effects can be obtained.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing immunoblotting and immunofluorescence staining results of a mouse corneal implant without rejection in normal, syngeneic, and allogenic corneal transplants;
FIG. 2 is a graph showing the effect of subconjunctival injection of 5mg/mL of BMS-P5, a drug that inhibits NETs formation, on immune rejection following a corneal graft.
Detailed Description
The invention provides an application of a preparation for inhibiting the formation of Neutrophil Extracellular Traps (NETs) in preparing a medicament for delaying and/or inhibiting immune rejection after cornea transplantation.
Formulations of the invention that inhibit the formation of NETs include an arginine deaminase 4 (PAD 4) inhibitor, and more preferably BMS-P5. The pharmacologically effective concentration of BMS-P5 of the present invention is preferably 4 to 60mg/mL, more preferably 5mg/mL. BMS-P5 according to the invention is preferably derived from Glpbio.
The invention also provides application of BMS-P5 in preparing anti-inflammatory drugs for inhibiting I-type IFN related channels.
In the embodiment of the invention, the intervention treatment of PAD4 inhibitor BMS-P5 is utilized to obviously inhibit the immune rejection of cornea transplantation, and the expression of a leukocyte surface active marker CD45 is obviously reduced after the BMS-P5 is injected, which proves that the subconjunctival injection of BMS-P5 of an allogeneic cornea transplantation rejection mouse can reduce the quantity of leukocytes in cornea implants and lighten the inflammatory reaction of cornea transplantation immune rejection; and the expression level of genes such as IFN-gamma, IL-1 beta, CXCL-10 and the like of the I-type IFN related inflammatory factors is reduced, namely BMS-P5 plays an anti-inflammatory role by inhibiting the I-type IFN related pathway.
The invention also provides a medicament for inhibiting cornea transplant immune rejection reaction, which comprises BMS-P5 with pharmacological effective concentration.
The medicament of the invention is preferably injection, and the pharmacological effective concentration is preferably 5mg/mL.
For further explanation of the present invention, the application of the PAD4 inhibitor provided in the present invention in preventing and treating immune rejection after cornea implantation is described in detail below with reference to the accompanying drawings and examples, which should not be construed as limiting the scope of the present invention.
EXAMPLE 1 increased expression of NETs in mouse corneal grafts following corneal transplantation with immune rejection
1. Male BALB/c mice of 8-10 weeks old were divided into the following 3 groups using a completely randomized block design:
A. mice not subjected to cornea transplantation served as a normal control group;
B. syngeneic allogeneic cornea transplant groups (donor is same week-old BALB/c mice);
C. allogeneic cornea transplant groups (donor is C57BL/6 mice of the same week), 6-8 of each group.
The penetrating cornea transplant model is established as follows: the mice are anesthetized by intraperitoneal injection of 0.6% pentobarbital sodium, all mice are subjected to right eye operation, conjunctival sac is flushed with sterile physiological saline before operation, and the compound topiramate eye drops are fully mydriatic, and the whole operation is carried out under a microscope. Taking the central cornea of a donor mouse as a implant by using a 2.5mm trephine, removing the central cornea of a recipient mouse by 2mm to prepare an implant bed, and intermittently suturing 8 needles by using 11-0 nylon suture lines to fix the implant on the implant bed. After the operation, air is injected into the anterior chamber to form the anterior chamber, ofloxacin eye ointment is coated in conjunctival sac to prevent infection, and the eyelid of the recipient mouse is sutured by using a 10-0 nylon suture. The corneal suture was removed on postoperative day 7. The slit lamp microscope was observed 3 times a week, and mice with complications such as cataract, iris adhesion, anterior chamber hemorrhage, infection, etc. were removed and supplemented with corresponding animals.
2. On the 19 th day after surgery, mice were sacrificed, eyeballs of group a and group C mice were taken, 3 samples each, and OCT gel was embedded and frozen into sections with a section thickness of 7 μm. The OCT gel was washed off with PBS, blotted around the specimen with a histochemical pen, the specimen was fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton-X, and blocked with 5% BSA for 1h at room temperature. The slides were spun dry and incubated for 4H at room temperature with rabbit anti-mouse H3cit antibody (1:200) and rat anti-mouse Ly6G antibody (1:100). The primary antibody was washed off with PBS and Alexa Fluor 594 donkey anti-rabbit antibody (1:1000) and Alexa Fluor 488 sheep anti-rat antibody (1:500) were co-incubated at room temperature for 1h. After washing off the antibodies, nuclei were stained with DAPI solution and observed under a positive inverted microscope (a in fig. 1). It was found that H3cit expression was significantly increased in the stromal and endothelial layers of the C-group implant cornea. H3cit is a marker for NETs formation, and an increase in expression level indicates an increase in NETs expression, and the above results indicate an increase in NETs formation in the corneal implant when the corneal graft rejection reaction occurs.
3. On the 19 th day after operation, mice were sacrificed by cervical removal, 3 mice per group were each obtained, the whole cornea was cut off along the limbus and placed in a 1.5ml centrifuge tube, pre-chilled RIPA lysate and milling beads were added, PMSF protease inhibitor was added at 1% concentration, sample proteins were extracted by tissue milling and ultrasound, and the proteins were denatured by a metal bath at 95 ℃ for 10 min. Proteins were separated by SDS-PAGE gel electrophoresis, transferred to PVDF membrane, blocked with 5% BSA at room temperature for 2H, and incubated overnight at 4℃with the addition of rabbit anti-H3 cit antibody (1:1000) and rabbit anti- β -actin antibody (1:1000). The following day was incubated with goat anti-rabbit IgG-HRP secondary antibody at room temperature for 2h after TBST washing, developed with ECL luminescence kit, and exposed with Bio-Rad rapid luminescence photographic apparatus (FIG. 1B). The results show that the protein expression level of the group C is obviously increased compared with that of the group A and the group B by taking the beta-actin as an internal reference. Citrullinated histones are markers of NETs formation, and increased expression levels indicate increased NETs expression, thus the results indicate increased NETs expression in implants following corneal transplantation where immune rejection occurs.
EXAMPLE 2PAD4 inhibitor BMS-P5 intervention treatment significantly inhibited corneal graft immune rejection
1. Using a completely randomized block design, 8-10 week old male BALB/c mice were randomly drawn into the following 3 groups:
A. mice not subjected to cornea transplantation served as a normal control group;
B. allogeneic cornea transplant groups;
C. allogeneic cornea transplant + BMS-P5 groups of 6-8 animals each.
A penetrating cornea implant model was established as described above. The drug was injected subconjunctival at 4 days post-surgery, with group B PBS solution, group C BMS-P5 at 5mg/kg and frequency of 1/3 d for 4 total injections. After operation, observation and photographing are carried out under a slit lamp every 1 day (figure 2), three indexes including transparency, edema degree and neovascularization of the cornea implant are scored by referring to a Larkin scoring method, the total observation time is 30 days, the occurrence time of each group of rejection reactions is recorded, and a cornea implant rejection survival curve is drawn.
As a result, on the 14 th day after the operation, the allograft group cornea implant gradually appears obvious edema, turbidity, peeping of the pupil and rejection reaction, while the BMS-P5 injection group cornea implant still keeps transparent. The time to rejection of the corneal implants was significantly prolonged in the BMS-P5 injected group compared to the allograft group (FIG. 2). Because PAD4 plays a key role in the NET formation process, the PAD4 inhibitor BMS-P5 can effectively inhibit NETs formation, so that the survival rate of the cornea implant is improved, and the immune rejection of cornea transplantation is inhibited.
2. On day 19 post-surgery, 3 eyes were taken for each group, the remaining mice were continuously observed, and OCT gel was embedded and frozen into sections. The OCT gel was washed off with PBS, blotted around the specimen with a histochemical pen, the specimen was fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton-X, and blocked with 5% BSA for 1h at room temperature. The slides were spun dry and the rabbit anti-CD 45 antibody incubated overnight at 4 ℃. The next day the primary antibody was washed off, the nuclei stained with DAPI solution and observed under a positive inverted microscope. As a result, the expression of the C-group leukocyte surface active marker CD45 was found to be significantly reduced as compared with the B-group, indicating that subconjunctival injection of BMS-P5 in allogeneic cornea transplant rejection mice can reduce the number of leukocytes in cornea implants and reduce the inflammatory response of cornea transplant immune rejection.
3. On the 19 th day after operation, 3 mice are taken from each group, the whole cornea is cut along the limbus, the total RNA of each group of cornea tissues is extracted by a Trizol method, the concentration of the RNA is measured on a Nanodrop, the mRNA is reversely transcribed into cDNA by a reverse transcription kit, a reaction solution is prepared by referring to the instruction book of the PCR kit, and each group of cDNA is added, and fluorescent quantitative PCR detection is carried out by taking GAPDH as an internal reference.
The primer sequences were as follows:
GAPDH upstream primer (SEQ ID No. 1): 5'-GCCACCCAGAAGACTGTGGAT-3' the number of the individual pieces of the plastic,
downstream primer (SEQ ID NO. 2): 5'-GGAAGGCCATGCCAGTGA-3';
IFN-gamma upstream primer (SEQ ID NO. 3): 5'-TCTTGGGTTCTTACGGCTGTTAC-3' the number of the individual pieces of the plastic,
downstream primer (SEQ ID NO. 4): 5'-TGTCACTCTCCTCTTTCCAATTCC-3';
IL-1. Beta. Upstream primer (SEQ ID NO. 5): 5'-CTTTCCCGTGGACCTTCCA-3' the number of the individual pieces of the plastic,
downstream primer (SEQ ID NO. 6): 5'-CTCGGAGCCTGTAGTGCAGTT-3';
CXCL-10 upstream primer (SEQ ID NO. 7): 5'-GGTCCGCTGCAACTGCAT-3' the number of the individual pieces of the plastic,
downstream primer (SEQ ID NO. 8): 5'-GGATTCAGACATCTCTGCTCATCA-3'.
By 2 —△△CT The method is used for relative quantitative standardization of data. The results show that the expression level of the I-type IFN related inflammatory factors IFN-gamma, IL-1 beta, CXCL-10 and the like is reduced. The results show that the expression level of inflammatory factor genes of the cornea implant in the BMS-P5 injection group is obviously lower than that in the allograft group, and the BMS-P5 plays an anti-inflammatory role by inhibiting the I-type IFN related pathway.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (3)
- Use of bms-P5 in the preparation of a medicament for delaying and/or inhibiting immune rejection following a corneal transplantation procedure.
- 2. The use according to claim 1, wherein the pharmacologically effective concentration of BMS-P5 is 4-60 mg/ml.
- 3. The use according to claim 2, characterized in that the pharmacologically effective concentration of BMS-P5 is 5mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310088670.2A CN116492462B (en) | 2023-02-03 | 2023-02-03 | Application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310088670.2A CN116492462B (en) | 2023-02-03 | 2023-02-03 | Application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116492462A CN116492462A (en) | 2023-07-28 |
| CN116492462B true CN116492462B (en) | 2023-09-22 |
Family
ID=87317262
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310088670.2A Active CN116492462B (en) | 2023-02-03 | 2023-02-03 | Application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116492462B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113244233A (en) * | 2021-06-23 | 2021-08-13 | 复旦大学附属眼耳鼻喉科医院 | Application of piperlongumine in preparation of medicine for preventing and treating corneal transplantation immunological rejection |
| CN113633642A (en) * | 2021-08-26 | 2021-11-12 | 山东第一医科大学附属青岛眼科医院(山东省眼科研究所、青岛眼科医院) | Application of ABT-263 in preparation of medicine for inhibiting corneal transplantation immune rejection |
| CN115066438A (en) * | 2020-01-31 | 2022-09-16 | 阿维拉治疗公司 | ASGPR binding compounds for degrading extracellular proteins |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210042932A (en) * | 2018-08-08 | 2021-04-20 | 브리스톨-마이어스 스큅 컴퍼니 | Substituted benzimidazole as a PAD4 inhibitor |
| WO2020055597A1 (en) * | 2018-09-13 | 2020-03-19 | The Children's Hospital Of Philadelphia | Net stabilization as a therapy for sepsis |
-
2023
- 2023-02-03 CN CN202310088670.2A patent/CN116492462B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115066438A (en) * | 2020-01-31 | 2022-09-16 | 阿维拉治疗公司 | ASGPR binding compounds for degrading extracellular proteins |
| CN113244233A (en) * | 2021-06-23 | 2021-08-13 | 复旦大学附属眼耳鼻喉科医院 | Application of piperlongumine in preparation of medicine for preventing and treating corneal transplantation immunological rejection |
| CN113633642A (en) * | 2021-08-26 | 2021-11-12 | 山东第一医科大学附属青岛眼科医院(山东省眼科研究所、青岛眼科医院) | Application of ABT-263 in preparation of medicine for inhibiting corneal transplantation immune rejection |
Non-Patent Citations (3)
| Title |
|---|
| A novel peptidylarginine deiminase 4 (PAD4) inhibitor BMS-P5 blocks formation of neutrophil extracellular traps and delays progression of multiple myeloma;Marina Li等;《Mol Cancer Ther.》;第19卷(第7期);第1530-1538页 * |
| PAD4 takes charge during neutrophil activation: Impact of PAD4 mediated NET formation on immune-mediated disease;Xiaosong Liu等;《J Thromb Haemost.》;第19卷;第1607-1617页 * |
| 角膜移植免疫排斥反应防治的研究进展;李琦等;《国际眼科杂志》;第6卷(第5期);第1126-1129页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116492462A (en) | 2023-07-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hos et al. | Immune reactions after modern lamellar (DALK, DSAEK, DMEK) versus conventional penetrating corneal transplantation | |
| Diamond et al. | Lensectomy and vitrectomy for complicated cataract secondary to uveitis | |
| Park et al. | Retroprosthetic membrane: a complication of keratoprosthesis with broad consequences | |
| Chen et al. | Glaucoma after ocular surgery or trauma: the role of infiltrating monocytes and their response to cytokine inhibitors | |
| EP3808352A1 (en) | Use of salidroside and derivative thereof in preparation of inhibitor medicament for diseases of ophthalmic fibrosis caused by abnormalities of extracellular matrix proteins | |
| Črnej et al. | Effect of penetrating keratoplasty and keratoprosthesis implantation on the posterior segment of the eye | |
| KR20180028890A (en) | Pharmaceutical Composition for Treating Macular Degeneration Containing mTOR Inhibitor | |
| RU2357743C1 (en) | Medication, method of obtaining medication, and method of treatment for cornea edema and other early bullous keratopathy manifestations | |
| CN116492462B (en) | Application of PAD4 inhibitor in preventing and treating immune rejection after cornea transplantation | |
| RU2465872C1 (en) | Method of prevention and treatment of excessive scarring after anti-glaucoma operation | |
| Fineman et al. | Topical Dorzolamide-lnduced Hypotony and Ciliochoroidal Detachment in Patients With Previous Filtration Surgery | |
| RU2369368C1 (en) | Method for prevention of inter-tissue adhesions after filtrating antiglaucomatous operation | |
| CN113559121B (en) | Medicine for treating cornea transplantation immune rejection | |
| RU2730975C1 (en) | Method of treating endothelial-epithelial dystrophy of cornea | |
| JPH08503968A (en) | Composition containing growth factor and antimetabolite | |
| US10568931B2 (en) | Pharmaceutical composition comprising ginseng extracts for prevention and treatment of ophthalmological diseases | |
| Langner-Wegscheider et al. | Surgical management of severe complications arising from uveitis in juvenile idiopathic arthritis | |
| Bao et al. | Effect of an MG132-sustained drug delivery capsular ring on the inhibition of posterior capsule opacification in a rabbit model | |
| RU2353341C2 (en) | Therapy of recurrent aching bullous keratopathy following phototherapeutic glaucoma eye keratectomy | |
| RU2421191C1 (en) | Method of treating dystrophic diseases of cornea | |
| RU2560390C1 (en) | Method of urgent treatment of destructive keratopathy | |
| RU2303457C1 (en) | Method for applying enzymotherapy | |
| RU2544306C2 (en) | Method of treating bullous keratopathy | |
| Doroodgar et al. | and Mohammad Ali Javadi | |
| Sheha | Update on Modulating Wound Healing in Trabeculectomy |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |