CN109419789B - Compound for treating and/or preventing immune disorder diseases and application thereof - Google Patents
Compound for treating and/or preventing immune disorder diseases and application thereof Download PDFInfo
- Publication number
- CN109419789B CN109419789B CN201710771752.1A CN201710771752A CN109419789B CN 109419789 B CN109419789 B CN 109419789B CN 201710771752 A CN201710771752 A CN 201710771752A CN 109419789 B CN109419789 B CN 109419789B
- Authority
- CN
- China
- Prior art keywords
- dendritic cells
- lymph nodes
- immune
- mice
- treating
- 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
- 150000001875 compounds Chemical class 0.000 title claims abstract description 14
- 208000026278 immune system disease Diseases 0.000 title abstract description 13
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title abstract description 12
- 210000001165 lymph node Anatomy 0.000 claims abstract description 58
- 210000004443 dendritic cell Anatomy 0.000 claims abstract description 42
- 102000018822 Retinal Dehydrogenase Human genes 0.000 claims abstract description 37
- 108010027691 Retinal dehydrogenase Proteins 0.000 claims abstract description 37
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 claims abstract description 26
- 102100022338 Integrin alpha-M Human genes 0.000 claims abstract description 26
- 101001046687 Homo sapiens Integrin alpha-E Proteins 0.000 claims abstract description 25
- 102100022341 Integrin alpha-E Human genes 0.000 claims abstract description 24
- 150000003839 salts Chemical class 0.000 claims abstract description 5
- 230000000968 intestinal effect Effects 0.000 claims description 15
- 230000005296 lymph node development Effects 0.000 claims description 11
- 230000001737 promoting effect Effects 0.000 claims description 3
- 208000007433 Lymphatic Metastasis Diseases 0.000 claims 1
- 230000002093 peripheral effect Effects 0.000 abstract description 13
- 201000010099 disease Diseases 0.000 abstract description 8
- 238000002474 experimental method Methods 0.000 abstract description 8
- 210000000987 immune system Anatomy 0.000 abstract description 7
- 208000004232 Enteritis Diseases 0.000 abstract description 3
- 230000036737 immune function Effects 0.000 abstract description 3
- 206010025135 lupus erythematosus Diseases 0.000 abstract description 3
- 201000006417 multiple sclerosis Diseases 0.000 abstract description 3
- 230000033228 biological regulation Effects 0.000 abstract description 2
- 230000002708 enhancing effect Effects 0.000 abstract description 2
- 230000002265 prevention Effects 0.000 abstract 1
- 230000001105 regulatory effect Effects 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 25
- 150000002632 lipids Chemical class 0.000 description 25
- 241000222178 Candida tropicalis Species 0.000 description 17
- 210000004027 cell Anatomy 0.000 description 16
- 210000001035 gastrointestinal tract Anatomy 0.000 description 16
- 239000000284 extract Substances 0.000 description 14
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 238000000034 method Methods 0.000 description 12
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 11
- 238000011740 C57BL/6 mouse Methods 0.000 description 11
- 241000233866 Fungi Species 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 10
- 239000002621 endocannabinoid Substances 0.000 description 9
- 210000004698 lymphocyte Anatomy 0.000 description 9
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 7
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 206010027476 Metastases Diseases 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 241000223230 Trichosporon Species 0.000 description 6
- 238000000684 flow cytometry Methods 0.000 description 6
- 230000028993 immune response Effects 0.000 description 6
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 6
- 239000007791 liquid phase Substances 0.000 description 6
- 230000009401 metastasis Effects 0.000 description 6
- 150000001200 N-acyl ethanolamides Chemical class 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000004811 liquid chromatography Methods 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 210000003462 vein Anatomy 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000003651 drinking water Substances 0.000 description 4
- 235000020188 drinking water Nutrition 0.000 description 4
- -1 endocannabinoid AEA analogs Chemical class 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000010992 reflux Methods 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 229940121375 antifungal agent Drugs 0.000 description 3
- 229930003827 cannabinoid Natural products 0.000 description 3
- 239000003557 cannabinoid Substances 0.000 description 3
- 230000002950 deficient Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- RFHAOTPXVQNOHP-UHFFFAOYSA-N fluconazole Chemical compound C1=NC=NN1CC(C=1C(=CC(F)=CC=1)F)(O)CN1C=NC=N1 RFHAOTPXVQNOHP-UHFFFAOYSA-N 0.000 description 3
- 229960004884 fluconazole Drugs 0.000 description 3
- 230000005012 migration Effects 0.000 description 3
- 238000013508 migration Methods 0.000 description 3
- 210000001986 peyer's patch Anatomy 0.000 description 3
- 238000005191 phase separation Methods 0.000 description 3
- 229920002307 Dextran Polymers 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 230000012292 cell migration Effects 0.000 description 2
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 230000002538 fungal effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 210000003563 lymphoid tissue Anatomy 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 208000029691 metastatic malignant neoplasm in the lymph nodes Diseases 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- NARAPOUIUKBGJL-GKFVBPDJSA-N (5z,8z,11z,14z)-n-propan-2-ylicosa-5,8,11,14-tetraenamide Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)NC(C)C NARAPOUIUKBGJL-GKFVBPDJSA-N 0.000 description 1
- 206010002556 Ankylosing Spondylitis Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- JVRTXNHWLKNPST-XVSDJDOKSA-N C(CC)N.C(CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC)(=O)O Chemical compound C(CC)N.C(CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC)(=O)O JVRTXNHWLKNPST-XVSDJDOKSA-N 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 102000008072 Lymphokines Human genes 0.000 description 1
- 108010074338 Lymphokines Proteins 0.000 description 1
- 101710139349 Mucosal addressin cell adhesion molecule 1 Proteins 0.000 description 1
- 102100028793 Mucosal addressin cell adhesion molecule 1 Human genes 0.000 description 1
- YLFLZSQBPHMIBS-GKFVBPDJSA-N N-propyl arachidonoyl amine Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)NCCC YLFLZSQBPHMIBS-GKFVBPDJSA-N 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 206010039438 Salmonella Infections Diseases 0.000 description 1
- 241000607762 Shigella flexneri Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241001502500 Trichomonadida Species 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000035605 chemotaxis Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 230000007412 host metabolism Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 210000005204 intestinal dendritic cell Anatomy 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000000207 lymphocyte subset Anatomy 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 229960000282 metronidazole Drugs 0.000 description 1
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 description 1
- 238000010232 migration assay Methods 0.000 description 1
- 230000001617 migratory effect Effects 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 210000001640 nerve ending Anatomy 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000005305 organ development Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 229930002330 retinoic acid Natural products 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 206010039447 salmonellosis Diseases 0.000 description 1
- 210000005212 secondary lymphoid organ Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 230000007847 structural defect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C([O-])=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 210000000264 venule Anatomy 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Images
Classifications
-
- 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
- A61K31/16—Amides, e.g. hydroxamic acids
-
- 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
- A61K31/16—Amides, e.g. hydroxamic acids
- A61K31/164—Amides, e.g. hydroxamic acids of a carboxylic acid with an aminoalcohol, e.g. ceramides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C233/00—Carboxylic acid amides
- C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C233/02—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals
- C07C233/09—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having nitrogen atoms of carboxamide groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals with carbon atoms of carboxamide groups bound to carbon atoms of an acyclic unsaturated carbon skeleton
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Physical Education & Sports Medicine (AREA)
- Rheumatology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Pain & Pain Management (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一种治疗和/或预防免疫紊乱疾病的化合物及其应用。本发明所提供的应用具体为式I或式II或式III所示化合物或其可药用盐在如下任一中的应用：(A)制备用于预防和/或治疗免疫紊乱相关疾病的产品；(B)制备用于调节外周免疫系统功能的产品；(C)预防和/或治疗免疫紊乱相关疾病；(D)调节外周免疫系统功能所示。实验证明，三个化合物均能够驱动肠道来源的树突状细胞(表达CD103、CD11b以及视黄醛脱氢酶)向淋巴结转移。本发明对于预防和/或治疗免疫紊乱相关疾病如风湿，红斑狼疮，肠炎，多发性硬化，并增强机体免疫功能具有重要意义。
The invention discloses a compound for treating and/or preventing immune disorders and its application. The application provided by the present invention is specifically the application of a compound represented by formula I or formula II or formula III or a pharmaceutically acceptable salt thereof in any of the following: (A) preparing a product for preventing and/or treating immune disorder-related diseases (B) preparation of products for regulating the function of peripheral immune system; (C) prevention and/or treatment of diseases related to immune disorders; (D) as shown in the regulation of peripheral immune system function. Experiments showed that all three compounds were able to drive gut-derived dendritic cells (expressing CD103, CD11b and retinal dehydrogenase) to metastasize to lymph nodes. The present invention is of great significance for preventing and/or treating immune disorder related diseases such as rheumatism, lupus erythematosus, enteritis, multiple sclerosis, and enhancing the immune function of the body. 
Description
Technical Field
The invention belongs to the technical field of biology, and relates to a compound for treating and/or preventing immune disorder diseases and application thereof.
Background
Microorganisms enter the animal body from skin, oral cavity, respiratory tract, etc., and they evolve together with the host to influence each other and inhibit and regulate the functions of the other. The research on the regulation and control mechanism between the symbiotic bacteria and the host has great significance to human health. Of the most concern to the immunologist is the effect of the gut flora on host metabolism and infection resistance. One of the issues is how the symbiotic bacteria regulate host immune function. Most literature studies focus on local immune regulation in the gut. This may be in the iceberg corner alone, and the effect of the commensal bacteria on the immune system may not be more than sufficient in the gut. For example, sterile animals have significant peripheral immunodeficiency, manifested primarily by hypoplasia of the systemic secondary lymphoid organs (i.e., lymph nodes). Since lymph nodes are the origin of immune responses, how symbionts regulate these widely distributed immune organs remotely has also become an important issue. However, to date, the association of these commensals with the development of the peripheral immune system has not been elucidated.
Lymph node development is a delicate and complex process. At the time of the embryo, the primordia of the lymph nodes appear in epithelial cell clusters. Upon stimulation with retinoic acid from nearby nerve endings, lymphoid tissue-inducing cells (LTi) begin to initiate the development of primitive lymphoid structures. After the mouse is born, LTi cells no longer remain, while peripheral lymph nodes continue to become larger and contain an increasing number of cells. One to two weeks after birth, the diafiltered lymphocytes form distinct T and B cell zones, almost identical to the lymph nodes of adult mice. In contrast, lymph node development in sterile animals was completely halted after birth. The developmental problems described above have led to a deficiency in the immune response. The immune response of the sterile mouse is weak after the sterile mouse is infected by intestinal pathogenic bacteria Shigella flexneri. Symptoms of salmonella infection are also more severe in germ-free mice. As a major site of immune response, it is conceivable that structural defects in lymph nodes may lead to a disturbance in the immune response. Then how do intestinal colonies initiate lymph node development after birth? Is the species of bacteria in the intestinal tract, which active ingredient acts?
Disclosure of Invention
The invention aims to provide a compound for treating and/or preventing immune disorder diseases and application thereof.
The invention also provides the application of the compound shown in the formula I, the formula II or the formula III or the pharmaceutically acceptable salt thereof in any one of the following (A) to (D), which belongs to the protection scope of the invention:
(A) preparing a product for preventing and/or treating diseases related to immune disorders;
(B) preparing a product for modulating peripheral immune system function;
(C) preventing and/or treating diseases related to immune disorder;
(D) modulating peripheral immune system function;
the invention also provides the use of a compound of formula I or formula II or formula III (supra) or a pharmaceutically acceptable salt thereof in any one of the following (E) to (H):
(E) preparing a product for driving the migration of gut-derived dendritic cells to lymph nodes;
(F) preparing a product for promoting lymph node development and/or lymph node function;
(G) driving gut-derived dendritic cells to metastasize to lymph nodes;
(H) promote lymph node development and/or promote lymph node function.
The invention also claims a product having at least one of the functions shown in the following (a) to (d), wherein the active ingredient is a compound shown in formula I, formula II or formula III (the same as above) or a pharmaceutically acceptable salt thereof; (a) preventing and/or treating diseases related to immune disorder; (b) modulating peripheral immune system function; (c) driving gut-derived dendritic cells to metastasize to lymph nodes; (d) promote lymph node development and/or promote lymph node function.
In the present invention, all the immune disorder-related diseases mentioned above can be autoimmune diseases such as rheumatism, lupus erythematosus, enteritis, multiple sclerosis, ankylosing spondylitis and the like, and excessive immune inflammatory responses.
In the present invention, all of the above-mentioned gut-derived dendritic cells are specifically gut-derived dendritic cells expressing CD103, CD11b and retinal dehydrogenase.
In one embodiment of the present invention, all of the aforementioned methods for driving metastasis of gut-derived dendritic cells to lymph nodes are specifically methods for driving metastasis of gut-derived dendritic cells to mesenteric lymph nodes and/or non-mesenteric peripheral lymph nodes.
Experiments prove that the compounds shown in the formula I and the formula II which are further extracted from the candida tropicalis lipid extract have similar structures with endogenous cannabinoid AEA (formula III), and all the compounds can drive intestinal dendritic cells (expressing CD103, CD11b and retinal dehydrogenase) to transfer to lymph nodes. Therefore, in practical application, it is expected that the intensity and type of immune response, and the number and function of each immune cell subtype can be adjusted by orally ingesting and injecting the compounds (endogenous cannabinoids derived from intestinal fungi can be chemically synthesized or used), so as to achieve the purposes of treating and/or preventing diseases related to immune disorders, such as rheumatism, lupus erythematosus, enteritis, multiple sclerosis and enhancing immune functions (such as immunotherapy and vaccination).
Drawings
FIG. 1 shows lymphocytes isolated from lymph nodes of adult normal mice, which are labeled and injected into normal (SPF) or sterile (GF) mice via the tail vein.
FIG. 2 shows lymph nodes of adult sterile mice lacking RALDH-positive DC cells.
FIG. 3 shows that the intestinal microorganisms are produced by the enzyme retinaldehyde dehydrogenase RALDH+Dendritic cells regulate lymph node development.
FIG. 4 shows CD103+CD11b+RALDH+Analysis of DC-like cell sources.
FIG. 5 shows intestinal CD103 driven by Candida tropicalis+CD11b+RALDH+The dendritic cells metastasize to lymph nodes. In the figure, RALDH at the ordinate+CD103+I.e. to CD103+CD11b+RALDH+A dendritic cell.
FIG. 6 shows that lipid extracts of Candida tropicalis are able to drive BMDC metastasis to lymph nodes.
FIG. 7 shows the results of liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of the ninth fragment obtained by liquid chromatography of lipid extract of Candida tropicalis.
Figure 8 shows that some molecular structures in the ninth paragraph (upper, middle) are very close to endocannabinoids (lower, AEA).
FIG. 9 shows that AEA and fungal fat extracts are able to strongly drive CD103 compared to other endocannabinoids+CD11b+RALDH+Dendritic cell migration.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 endocannabinoid AEA analogs from Candida tropicalis can drive intestinal DC cell metastasis to lymph nodes
First, lymphocyte to lymph node reflux in sterile mice were deficient
Lymphocytes isolated from lymph nodes of adult normal C57BL/6 mice, labeled with CFSE, were injected into normal (SPF) or sterile (GF) C57BL/6 mice (at a dose of 2X 10 per mouse) via tail vein6Cells) were isolated 24 hours later for various lymph nodes, and either sectioned for fluorescence imaging or cell suspension taken for analysis by flow cytometry.
The results are shown in FIG. 1. Left: reflux of injected lymphocytes to Inguinal Lymph Node (iLN), mesenteric lymph node (mLN) and spleen (spl), and analysis of lymphocyte subset reflux to iLN (bottom left). And (3) right: expression of MAdCAM-1 and PNAd on High Endothelial Venules (HEVs) within iLN in GF mice (converted) after caging with SPF, and SPF. This experiment showed that lymphocyte reflux to the lymph nodes was deficient in germ free mice due to the absence of expression of PNAd.
II, adult sterile mouse lymph node deletion RALDH positive DC cells
Lymph nodes from 5-week normal (SPF) and sterile (GF) C57BL/6 mice were stained and compared to see the presence or absence of RALDH-positive DC cells in both.
The results are shown in FIG. 2. The method comprises the following steps: adult sterile mice have a depletion in their lymph nodes of a class of dendritic cells of retinal dehydrogenase (RALDH) compared to normal mice. After the SPF mice are bred in cages, RALDH positive cells appear in peripheral lymph nodes of the sterile mice. The following: and (4) comprehensive data analysis.
Thirdly, the intestinal microorganisms are obtained by RALDH+Dendritic cells regulate lymph node development
Isolation of RALDH from lymph nodes of SPF grade C57BL/6 mice+Dendritic cells and RALDH-Dendritic cells were injected via tail vein into sterile (GF) C57BL/6 mice (injection dose 2X 10)5Cells), the lymph nodes were sectioned seven days later and analyzed by flow cytometry.
The results are shown in FIG. 3. The method comprises the following steps: RALDH isolated from lymph nodes of SPF mice+Dendritic cells or RALDH-Dendritic cells, injected into sterile mice via the tail vein, cause changes in lymph node volume and imaging of mature T, B regions. The following: lymph node size, and statistics of the areas of zone B and zone T. This experiment demonstrated that intestinal microorganisms were produced by RALDH+This class of dendritic cells regulates lymph node development.
IV, CD103+CD11b+RALDH+Analysis of DC-like cell origin
The effect of gut symbionts on immune organ development may not be direct. Within the lamina propria of the intestinal tract is a group of unconventional dendritic cells which are capable of expressing CD103, CD11b and retinal dehydrogenase (RALDH). They can come into contact with lymphocytes, and this process has a great influence on the fate of these lymphocytes. Apparently, there appears to be no connection between lymph node development and the above processes. Earlier stage work in our laboratory found that intestinal colonies could be timely released while the lymphoid tissue of newborn mice induced cell disappearanceNow, a CD103 gene appears in the lymph nodes+CD11b+DC-like cells. These cells isolated from normal mice, when injected into sterile mice via tail vein, promote the development of the latter lymph nodes and the entry of a large number of lymphocytes into the lymph nodes. These dendritic cells highly express retinal dehydrogenase and are capable of expressing a substance called peripheral lymphokine on the vascular epithelium at the entrance to the lymph node, which can also be considered as a marker for address localization. T and B cells develop lymph nodes by recognizing peripheral lymphokines entering the lymph nodes. This process is particularly evident at the time of birth of the mice. However, in adult mice, the lymph nodes still have a small number of such dendritic cells, which maintain long-term lymph node homeostasis. In vitamin a deficient mice, dendritic cells such as lymph nodes disappear, causing structural destruction. We have shown by means of a marker that such dendritic cells originate from the gut, and that the unconventional dendritic cells in the lamina propria of the gut (which express CD103, CD11b and retinal dehydrogenase) are of the same type as we have said above.
The specific method comprises the following steps: neonatal C57BL/6 mice were gavaged with FITC Dextran (FITC-Dextran 2000KD (Sigma)0.3 mg per g body weight) and after 6 hours the proportion of CD11b and CD103 single positive, Double Negative (DN) and Double Positive (DP) dendritic cells in peripheral lymph nodes was determined by flow cytometry.
As shown in FIG. 4, the FITC signal was strongest in CD11b and CD103 double positive dendritic cells, which were found to originate from the intestinal tract.
Fifth, candida tropicalis drives intestinal CD103+CD11b+RALDH+Metastasis of dendritic cells to lymph nodes
Adult SPF grade C57BL/6 mice were mixed with either mixed antibiotics (antibacterial only and antifungal, 1g/L ampicillin, 1g/L neomycin, 1g/L metronidazole and 0.5g/L vancomycin, concentrations expressed as final concentrations in drinking water, Sigma) or the antifungal agent fluconazole (Sigma) (final concentrations in drinking water of 0.5g/L, Sigma) in drinking water and three weeks later lymph nodes were taken and subjected to flow cytometry analysis of mouse mesenteric lymph nodes (mLN) and peyer's patchesCD103 in Baker (PP)+CD11b+RALDH+Percentage of dendritic cells.
Adult normal C57BL/6 mice were individually gavaged with three major intestinal fungi cultured, Candida tropicalis, Saccharomyces cerevisiae and Trichosporon trichotomum (10 per strain per mouse)8CFU)24 hours later, the lymph nodes were taken for flow cytometry analysis of CD103 in the mouse Inguinal (iLN) and mesenteric (mLN) lymph nodes+CD11b+RALDH+Percentage of dendritic cells. In the same manner, the experiment was performed on newborn C57BL/6 mice and sterile C57BL/6 mice.
The results are shown in FIG. 5. Upper left: adult SPF mice were mixed with a mixture of antibiotic or antifungal fluconazole (fluconazole) in drinking water, and three weeks later, mesenteric lymph nodes (mLN) and Peyer's Patch (PP) with CD103+CD11b+RALDH+Percentage of dendritic cells. Upper right: adult normal mice were CD103 in Inguinal (iLN) and mesenteric (mLN) lymph nodes after 24 hours of gavage with cultured Candida tropicalis (Candida tropicalis), Saccharomyces cerevisiae (Saccharomyces cerevisiae), and Trichosporon (Trichosporon)+CD11b+RALDH+Percentage of dendritic cells. Left lower: similar to the upper right, results for newborn mice. Right lower: similar to the results for the upper right and lower left, adult germ-free mice. It can be seen that Candida tropicalis (C tropicalis) in intestinal microorganisms drives CD103+CD11b+RALDH+Metastasis of dendritic cells to lymph nodes.
Lipid extract of Candida tropicalis capable of driving intestinal CD103+CD11b+RALDH+Metastasis of dendritic cells to lymph nodes
Ribonucleic acid, protein and lipid of fungi and bacteria (the fungi include Candida tropicalis, Saccharomyces cerevisiae and Trichosporon trichomonad, and the bacteria are Escherichia coli) are obtained by separation and extraction. After reinjection of these isolates into mice, only the Candida tropicalis lipids were able to drive the transfer of dendritic cells to lymph nodes. The method comprises the following specific steps:
the chloroform-methanol extraction method is used for extracting lipids of fungi and bacteria, wherein the fungi include Candida tropicalis (Candida tropicalis), Saccharomyces cerevisiae (Saccharomyces cerevisiae) and Trichosporon (Trichosporon), and the bacteria include Escherichia coli (E coli). Ultrasonically crushing fungi or bacteria in chloroform-methanol (volume ratio of 2:1), centrifuging, washing with water to remove methanol, and blowing dry chloroform with nitrogen to obtain lipid extract.
The lipid extracts of the fungi and bacteria obtained above were subjected to liquid chromatography analysis, as follows: a chromatographic column: the diameter is 1.5cm, and the length is 30 cm; the filler is silica gel (200-300 mesh, particle size 45-75 μm, pore diameter 40-70A, specific surface area 400-600 m)2Per gram, pore volume 0.60-0.85 ml/gram), filled at atmospheric pressure. Lipids extracted from 5g (wet weight) of fungi or bacteria were loaded with 100% chloroform (30ml) and then treated with chloroform: methanol was eluted at 10:0, 8:2, 6:4, 5:5, 3:7, 0:10 volumes, 30ml each for the first 5 gradients, each divided equally into two front and back sections, and 45ml for the last gradient (i.e. gradient No. 6), divided equally into three sections. Thereby forming 1 to 13 paragraphs. At room temperature, the samples were allowed to fall freely and all samples were blown dry with nitrogen.
The lipid extract was separated into fractions by liquid phase separation, each fraction was dissolved in the same volume of DMSO as the total non-fractionated lipids, and subjected to Transwell assay (lipid was added to the lower chamber, and the ratio of lipid to culture medium was 1:1000 to determine the activity of each fraction to attract migration of mouse bone marrow-derived dendritic cells (BMDC) (the chemotaxis of BMDC and CD103 was clear to those skilled in the art)+CD11b+RALDH+Dendritic cells were similar, and BMDC was used in place of CD103 in this experiment+CD11b+RALDH+The reason for dendritic cells is that: the latter is difficult to obtain sufficient cell number for mass analysis, and moreover, the latter is not very active in culture and is not suitable for long-term experiments).
The results are shown in FIG. 6, A: liquid chromatography spectra of various fungal lipids; b: several segments of tropical yeast induce the migration of dendritic cells in vitro, the ninth segment being the strongest. The relevant activities in the lipids of visible fungi can be concentrated and purified by the liquid phase.
Seventhly, structure determination of endocannabinoid AEA analogue from candida tropicalis
And (3) separating the C tropicalis lipid extract in a liquid phase for a ninth section, and performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, and performing automatic analysis (Full Scan mode) on data of a Waters QTOF mass spectrometer by Thermal X-caliber software to obtain the molecular composition of the lipid.
The results are shown in FIGS. 7 and 8.
In fig. 7, upper: c tropicalis lipid extract liquid phase separation ninth section liquid chromatography analysis; liquid chromatography results of N-arachidonic acid aminoethanol (AEA) standard. The following: AEA mass spectrometry results; c tropicalis lipid extract liquid phase separation of AEA homologues extracted from the LC-MS/MS results in the ninth stage.
In fig. 8, upper, middle: some molecular structures in the ninth paragraph (shown as formula I and formula II, respectively). The following: endocannabinoids (AEA is shown in formula III)) molecular structure.
The results show that: c tropicalis lipid extract the ninth stage contained the endogenous cannabinoid AEA homologue by liquid phase isolation. According to the LC-MS/MS results, the AEA homologue was closest to arachidonic acid N-propylamine (N-propyl arachidonoyl amine) or arachidonic acid isopropylamine (N-isopropyl arachidonoyl amine).
Eight, endocannabinoid AEA analogs from Candida tropicalis capable of driving intestinal CD103+CD11b+RALDH+Metastasis of dendritic cells to lymph nodes
Different endocannabinoids and C tropicalis lipid extracts were injected intra-abdominally into 4-week-old SPF grade C57BL/6 mice. DMSO stocks of C tropicalis lipid and cannabinoid were dissolved in PBS buffer for injection. DMSO at the same dose served as a blank. The treatment time was one hour. Flow cytometry detection of migratory DCs (i.e., CD 103) in mesenteric lymph nodes (mLNs)+CD11b+RALDH+Dendritic cells). The amount of C tropicalis lipid was 250. mu.g per mouse dissolved in DMSO fraction.
The results are shown in FIG. 9, where it can be seen that: lipid extracts of AEA and C tropicalis are able to drive CD103 more strongly than other endocannabinoids+CD11b+RALDH+Dendritic cell migration.
Claims (2)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710771752.1A CN109419789B (en) | 2017-08-31 | 2017-08-31 | Compound for treating and/or preventing immune disorder diseases and application thereof |
| PCT/CN2017/113538 WO2019041596A1 (en) | 2017-08-31 | 2017-11-29 | Compound for treating and/or preventing immune disorder disease and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710771752.1A CN109419789B (en) | 2017-08-31 | 2017-08-31 | Compound for treating and/or preventing immune disorder diseases and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109419789A CN109419789A (en) | 2019-03-05 |
| CN109419789B true CN109419789B (en) | 2021-09-21 |
Family
ID=65505340
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710771752.1A Active CN109419789B (en) | 2017-08-31 | 2017-08-31 | Compound for treating and/or preventing immune disorder diseases and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN109419789B (en) |
| WO (1) | WO2019041596A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114350616A (en) * | 2022-01-24 | 2022-04-15 | 深圳市先康达生命科学有限公司 | Immune cell and preparation method and application thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2192965C (en) * | 1994-07-11 | 2007-12-04 | David W. Pate | Anandamide analogue compositions and method of treating intraocular hypertension using same |
| WO2005023232A2 (en) * | 2003-09-04 | 2005-03-17 | Affibody Ab | Therapeutic, screening and diagnostic methods based on the effect of cb1 receptor modulation on lymphoproliferative disorders. |
| US20170112787A1 (en) * | 2015-05-08 | 2017-04-27 | The University Of Connecticut | Methods of treatment of inflammation of the gut |
| EP3095444A1 (en) * | 2015-05-20 | 2016-11-23 | Dublin City University | A method of treating peripheral inflammatory disease |
| WO2017066747A1 (en) * | 2015-10-16 | 2017-04-20 | Scythian Biosciences Inc. | Methods and compositions for treating gastrointestinal inflammation |
-
2017
- 2017-08-31 CN CN201710771752.1A patent/CN109419789B/en active Active
- 2017-11-29 WO PCT/CN2017/113538 patent/WO2019041596A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2019041596A1 (en) | 2019-03-07 |
| CN109419789A (en) | 2019-03-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Diling et al. | Immunomodulatory activities of a fungal protein extracted from Hericium erinaceus through regulating the gut microbiota | |
| US11980660B2 (en) | Edible vaccination against microbial pathogens | |
| Huang et al. | Myricetin possesses anthelmintic activity and attenuates hepatic fibrosis via modulating TGFβ1 and Akt signaling and shifting Th1/Th2 balance in Schistosoma japonicum-infected mice | |
| JP2021120383A (en) | Use of beta-1,3-glucan for modulating immune function and treating intestinal inflammation | |
| Lin et al. | Immunopathological effects of Agaricus blazei Murill polysaccharides against Schistosoma mansoni infection by Th1 and NK1 cells differentiation | |
| CN117165465A (en) | Lactobacillus plantarum strain for improving intestinal barrier function and application thereof | |
| WO2005040375A9 (en) | A fungal immuromodulatory protein produced by microorganisme and uses thereof | |
| Wang et al. | Litchi thaumatin-like protein induced the liver inflammation and altered the gut microbiota community structure in mice | |
| CN109419789B (en) | Compound for treating and/or preventing immune disorder diseases and application thereof | |
| JP4160532B2 (en) | Pharmaceutical composition for enhancing immunity and polya extract | |
| WO2018066676A1 (en) | Composition comprising organic selenium compound | |
| CN109420173B (en) | Application of GPR55 and its regulators in the prevention and treatment of immune system diseases | |
| CN109420174B (en) | Application of GPR18 and its regulators in the prevention and treatment of immune system diseases | |
| CN109419820B (en) | Application of candida tropicalis in preparation of products for preventing and treating immune system diseases | |
| JP6158541B2 (en) | Intestinal immunity regulator | |
| US12161662B2 (en) | Structural and functional characteristics of yeast-derived polysaccharide inducing Treg cell | |
| US11554144B2 (en) | Nanovesicles derived from enhydrobacter bacteria, and use thereof | |
| US12018337B2 (en) | Nano-vesicle derived from catenibacterium bacteria and use thereof | |
| Wu et al. | Major royal-jelly proteins intake modulates immune functions and gut microbiota in mice | |
| JP2004099613A (en) | Ganoderma lucidum spores for treatment of autoimmune diseases | |
| US20240408053A1 (en) | Enteritis-suppressing composition | |
| JPH03218466A (en) | Plant glycolipid positive in limulus test, immune function activator, immune function activator for animal, immune function inspecting drug, immune function inspecting drug for animal, non-pharmaceuticals, cosmetics, food, functional food, drinks, feed containing such glycolypid | |
| KR20130103468A (en) | Expanding bone marrow-derived immune regulatory cells and immune regulatory b cells by activating gpcr19 pathway in vivo or in vitro | |
| CN118021917A (en) | Pharmaceutical compound composition for preventing or treating Alzheimer disease and application thereof | |
| JP6436971B2 (en) | Composition for reducing CD4 and CD8 memory cells comprising ginseng fruit extract |
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 |