CN117503782A - Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis - Google Patents
Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis Download PDFInfo
- Publication number
- CN117503782A CN117503782A CN202311533828.9A CN202311533828A CN117503782A CN 117503782 A CN117503782 A CN 117503782A CN 202311533828 A CN202311533828 A CN 202311533828A CN 117503782 A CN117503782 A CN 117503782A
- Authority
- CN
- China
- Prior art keywords
- ginsenoside
- liver
- immune hepatitis
- use according
- hepatitis
- 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
- FVIZARNDLVOMSU-IRFFNABBSA-N ginsenoside C-K Chemical compound O([C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(CC[C@H]4C(C)(C)[C@@H](O)CC[C@]4(C)[C@H]3C[C@H]2O)C)(C)CC1)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O FVIZARNDLVOMSU-IRFFNABBSA-N 0.000 title claims abstract description 42
- 239000003814 drug Substances 0.000 title claims abstract description 22
- 208000006454 hepatitis Diseases 0.000 title claims abstract description 18
- 231100000283 hepatitis Toxicity 0.000 title claims abstract description 17
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 claims abstract description 11
- 102100039360 Toll-like receptor 4 Human genes 0.000 claims abstract description 11
- 206010003827 Autoimmune hepatitis Diseases 0.000 claims description 11
- 239000003826 tablet Substances 0.000 claims description 7
- 235000003140 Panax quinquefolius Nutrition 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 241000208340 Araliaceae Species 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 claims description 3
- 240000005373 Panax quinquefolius Species 0.000 claims description 3
- 235000008434 ginseng Nutrition 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 108010062580 Concanavalin A Proteins 0.000 claims description 2
- 241000180649 Panax notoginseng Species 0.000 claims description 2
- 235000003143 Panax notoginseng Nutrition 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000007910 chewable tablet Substances 0.000 claims description 2
- 239000007919 dispersible tablet Substances 0.000 claims description 2
- 239000002552 dosage form Substances 0.000 claims description 2
- 239000002662 enteric coated tablet Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 229940124531 pharmaceutical excipient Drugs 0.000 claims description 2
- 239000000725 suspension Substances 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims 1
- 229940068682 chewable tablet Drugs 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 239000004615 ingredient Substances 0.000 claims 1
- 230000011664 signaling Effects 0.000 claims 1
- 210000004185 liver Anatomy 0.000 abstract description 30
- 230000000694 effects Effects 0.000 abstract description 16
- 238000011282 treatment Methods 0.000 abstract description 10
- 230000014509 gene expression Effects 0.000 abstract description 9
- 210000005228 liver tissue Anatomy 0.000 abstract description 9
- 229940079593 drug Drugs 0.000 abstract description 7
- 229940089161 ginsenoside Drugs 0.000 abstract description 5
- 229930182494 ginsenoside Natural products 0.000 abstract description 5
- 206010067125 Liver injury Diseases 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 4
- 206010061218 Inflammation Diseases 0.000 abstract description 3
- 238000011161 development Methods 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 3
- 231100000234 hepatic damage Toxicity 0.000 abstract description 3
- 230000004054 inflammatory process Effects 0.000 abstract description 3
- 230000008818 liver damage Effects 0.000 abstract description 3
- 230000003908 liver function Effects 0.000 abstract description 3
- 230000006907 apoptotic process Effects 0.000 abstract description 2
- 210000005229 liver cell Anatomy 0.000 abstract description 2
- 230000002265 prevention Effects 0.000 abstract description 2
- 230000004792 oxidative damage Effects 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 14
- 230000002757 inflammatory effect Effects 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 210000000952 spleen Anatomy 0.000 description 6
- 238000002156 mixing Methods 0.000 description 5
- 230000001105 regulatory effect Effects 0.000 description 5
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 4
- 102000018745 NF-KappaB Inhibitor alpha Human genes 0.000 description 4
- 108010052419 NF-KappaB Inhibitor alpha Proteins 0.000 description 4
- 102000003929 Transaminases Human genes 0.000 description 4
- 108090000340 Transaminases Proteins 0.000 description 4
- 229940009098 aspartate Drugs 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 239000002934 diuretic Substances 0.000 description 4
- 230000026731 phosphorylation Effects 0.000 description 4
- 238000006366 phosphorylation reaction Methods 0.000 description 4
- 230000019491 signal transduction Effects 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 229940030606 diuretics Drugs 0.000 description 3
- 230000008595 infiltration Effects 0.000 description 3
- 238000001764 infiltration Methods 0.000 description 3
- 210000004969 inflammatory cell Anatomy 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 239000004375 Dextrin Substances 0.000 description 2
- 229920001353 Dextrin Polymers 0.000 description 2
- 206010016654 Fibrosis Diseases 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 102000002689 Toll-like receptor Human genes 0.000 description 2
- 108020000411 Toll-like receptor Proteins 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 235000019425 dextrin Nutrition 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000007901 soft capsule Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- RAQNTCRNSXYLAH-RFCGZQMISA-N (20S)-ginsenoside Rh1 Chemical compound O([C@@H]1[C@H]2C(C)(C)[C@@H](O)CC[C@]2(C)[C@H]2C[C@@H](O)[C@H]3[C@@]([C@@]2(C1)C)(C)CC[C@@H]3[C@@](C)(O)CCC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RAQNTCRNSXYLAH-RFCGZQMISA-N 0.000 description 1
- KWDWBAISZWOAHD-MHOSXIPRSA-N (2s,3r,4s,5s,6r)-2-[(2r,3r,4s,5s,6r)-4,5-dihydroxy-6-(hydroxymethyl)-2-[[(3s,5r,8r,9r,10r,12r,13r,14r,17s)-12-hydroxy-4,4,8,10,14-pentamethyl-17-(6-methylhepta-1,5-dien-2-yl)-2,3,5,6,7,9,11,12,13,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-3-yl]oxy]o Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1CC[C@]2(C)[C@H]3C[C@@H](O)[C@H]4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@@H]4C(=C)CCC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O KWDWBAISZWOAHD-MHOSXIPRSA-N 0.000 description 1
- WSWCOQWTEOXDQX-MQQKCMAXSA-M (E,E)-sorbate Chemical compound C\C=C\C=C\C([O-])=O WSWCOQWTEOXDQX-MQQKCMAXSA-M 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- 208000007848 Alcoholism Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- NJUXRKMKOFXMRX-RNCAKNGISA-N Ginsenoside Rg5 Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1CC[C@]2(C)[C@H]3C[C@@H](O)[C@H]4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@@H]4C(/C)=C/CC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O NJUXRKMKOFXMRX-RNCAKNGISA-N 0.000 description 1
- KWDWBAISZWOAHD-UHFFFAOYSA-N Ginsenoside Rk1 Natural products CC(C)=CCCC(=C)C1CCC(C2(CCC3C4(C)C)C)(C)C1C(O)CC2C3(C)CCC4OC1OC(CO)C(O)C(O)C1OC1OC(CO)C(O)C(O)C1O KWDWBAISZWOAHD-UHFFFAOYSA-N 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 206010022489 Insulin Resistance Diseases 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 206010027951 Mood swings Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 1
- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 108700012920 TNF Proteins 0.000 description 1
- 101150082427 Tlr4 gene Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 208000021017 Weight Gain Diseases 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- LMEKQMALGUDUQG-UHFFFAOYSA-N azathioprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC=NC2=C1NC=N2 LMEKQMALGUDUQG-UHFFFAOYSA-N 0.000 description 1
- 229960002170 azathioprine Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000003560 cancer drug Substances 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 230000007012 clinical effect Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000001882 diuretic effect Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- NJUXRKMKOFXMRX-AXUZFSSLSA-N ginsenoside Rg5 Natural products CC(=CCC=C(C)[C@H]1CC[C@]2(C)[C@@H]1[C@H](O)C[C@@H]3[C@@]4(C)CC[C@@H](O[C@H]5O[C@H](CO)[C@@H](O)[C@H](O)[C@H]5O[C@H]6O[C@H](CO)[C@@H](O)[C@H](O)[C@H]6O)C(C)(C)[C@@H]4CC[C@@]23C)C NJUXRKMKOFXMRX-AXUZFSSLSA-N 0.000 description 1
- NJUXRKMKOFXMRX-UHFFFAOYSA-N ginsenoside Rz1 Natural products CC(C)=CCC=C(C)C1CCC(C2(CCC3C4(C)C)C)(C)C1C(O)CC2C3(C)CCC4OC1OC(CO)C(O)C(O)C1OC1OC(CO)C(O)C(O)C1O NJUXRKMKOFXMRX-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 206010019692 hepatic necrosis Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 208000018191 liver inflammation Diseases 0.000 description 1
- 231100000149 liver necrosis Toxicity 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000989 no adverse effect Toxicity 0.000 description 1
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 1
- 235000006286 nutrient intake Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000004798 organs belonging to the digestive system Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000003617 peroxidasic effect Effects 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229950008882 polysorbate Drugs 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 229940069328 povidone Drugs 0.000 description 1
- 229960005205 prednisolone Drugs 0.000 description 1
- OIGNJSKKLXVSLS-VWUMJDOOSA-N prednisolone Chemical compound O=C1C=C[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 OIGNJSKKLXVSLS-VWUMJDOOSA-N 0.000 description 1
- 229960004618 prednisone Drugs 0.000 description 1
- XOFYZVNMUHMLCC-ZPOLXVRWSA-N prednisone Chemical compound O=C1C=C[C@]2(C)[C@H]3C(=O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 XOFYZVNMUHMLCC-ZPOLXVRWSA-N 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229940075554 sorbate Drugs 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 description 1
- 229940013618 stevioside Drugs 0.000 description 1
- 235000019202 steviosides Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 201000009032 substance abuse Diseases 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7032—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a polyol, i.e. compounds having two or more free or esterified hydroxy groups, including the hydroxy group involved in the glycosidic linkage, e.g. monoglucosyldiacylglycerides, lactobionic acid, gangliosides
-
- 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
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Epidemiology (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses application of ginsenoside CK in preparing medicines for treating immune hepatitis. Experiments prove that the ginsenoside CK can regulate the apoptosis, inflammatory reaction, oxidative damage and the like of liver cells of immune hepatitis for the first time, so that the liver damage is relieved, the normal operation of liver functions is effectively protected, and the CK also has a regulation and control effect on the expression of TLR4 isogenes in liver tissues, so that the protection effect of the ginsenoside CK on the liver is related to a TLR4/NF- κB signal path. The prevention and treatment effect and mechanism of the ginsenoside CK for the immune hepatitis are clear, and the ginsenoside has smaller side effect and higher safety. The invention provides a wider treatment range for the development of ginsenoside-action medicaments, provides a new and more effective treatment method for the treatment of immune hepatitis, and has good economic benefit and social influence.
Description
Technical Field
The invention relates to the technical field of biological pharmacy, in particular to application of ginsenoside CK in preparing medicines for preventing and treating immune hepatitis.
Background
The liver is a necessary digestive organ of the human body, is responsible for metabolism of various nutrient intakes, and is the place where various chemical reactions are most active. However, in recent years, the incidence of liver damage has increased due to drugs of abuse, alcoholism, and viral infection. Autoimmune hepatitis (AIH) is a severe immune-mediated liver disease of unknown etiology and pathogenesis, which can occur in children and adults of all ages. If not properly treated, AIH may develop cirrhosis or even hepatocellular carcinoma. The clinical symptoms of AIH are significant elevation of serum Alanine (ALT) or Aspartate (AST) aminotransferase levels, abnormally abundant inflammatory cell infiltration and liver necrosis in the liver, etc.
Currently, immunosuppressants-hormones are the most widely used therapeutic methods in the drug treatment of autoimmune hepatitis, including prednisone or prednisolone combined with azathioprine, and have good clinical effects. However, the side effects associated with these treatments are also more prevalent, such as weight gain, mood swings, and osteoporosis, which alert people to the urgent need to develop safer and more effective AIH therapies.
Among the many inflammatory regulatory signaling pathways, the NF- κB signaling pathway has a regulatory effect on the production of transcription factors, inflammatory cytokines, and even cell survival, whereas overexpression of Toll-like receptors (TLRs) activates NF- κB, thereby initiating an immune response. Thus, the TLR4/NF- κB pathway plays a critical role in the development and progression of immune related diseases.
In recent years, various plant chemicals extracted from natural products have been explored to demonstrate various biological activities such as antioxidant, anti-inflammatory, etc. Among them, ginsenoside is used as the main bioactive component in the Araliaceae plants such as ginseng, american ginseng, notoginseng, etc., and its biological activity has been widely verified in experimental level and clinical study in the past decades. Researches report that the ginsenoside CK has a strong protection effect on liver injury due to good anti-inflammatory and antioxidant activities. Based on the above, the technical proposal of the invention proposes that the ginsenoside CK improves the occurrence and development of autoimmune hepatitis by regulating inflammatory reaction mediated by TLR4/NF- κB channel.
Chinese patent No. CN202010525824.6 discloses application of ginsenoside CK in anti-colon cancer drugs, chinese patent No. CN202010525824.6 discloses application of ginsenoside CK in lipid-lowering and weight-losing drugs, chinese patent No. CN 201510493410.9 discloses application of ginsenoside CK and Rh1 monomers and compositions thereof in drugs for improving nonalcoholic fatty liver fibrosis and insulin resistance, and Chinese patent No. CN 201810482539.3 discloses a total ginsenoside composition, ginsenoside Rg5, ginsenoside Rk1 and ginsenoside CK which can be used for preparing diuretics or diuretics health care products, so that the technical problems of poor diuretic reaction and diuretics resistance in the prior art are solved. At present, reports of ginsenoside CK for treating immune hepatitis are not yet seen.
Disclosure of Invention
In view of the above, the invention discloses application of ginsenoside CK in preparing medicines for preventing and treating immune hepatitis.
Further, the ginsenoside CK is the main or only effective component in the medicine.
Further, the immune hepatitis is an autoimmune hepatitis induced by concanavalin A.
Further, the ginsenoside CK is obtained by converting Rb1 extracted from ginseng, pseudo-ginseng or American ginseng by snailase, and the purity of the ginsenoside CK is more than or equal to 95%.
Further, the medicine is a composition composed of ginsenoside CK, pharmaceutical excipients and other auxiliary components.
Further, the medicament is an oral preparation, and the dosage form comprises one of tablets, capsules, granules, suspensions and powder, wherein the tablets comprise common tablets, dispersible tablets, chewable tablets and enteric-coated tablets.
Further, the single dosage of the ginsenoside CK is 10-100mg/kg.
Further, the medicine is a medicine for improving immune hepatitis through a TLR4/NF- κB signaling pathway.
In the present invention, the preventive and/or therapeutic effect is achieved by one or more of improving the level of trans-hepatic amino acids, maintaining the normal structure of the liver, reducing inflammatory factors and inflammatory cell infiltration of the liver, and improving liver peroxidative damage.
Compared with the prior art, the invention has the following beneficial effects:
1. experiments prove that the ginsenoside CK pretreatment can improve ConA-induced mouse immune hepatitis, reduce the transaminase level and the expression of inflammatory factors, improve liver inflammatory cell infiltration, and relieve liver peroxidation damage and apoptosis, and the ginsenoside has definite effect on treating immune hepatitis and has no adverse effect.
2. The invention provides a wider choice for the application of the ginsenoside, also provides a new, safe and effective prevention and treatment mode for the treatment of immune hepatitis, and has good social and economic benefits.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the relative liver and spleen masses of mice after the end of the experiment and the body weight of the mice during the administration period;
FIG. 2 is a liver, spleen tissue morphology and H & E staining;
FIG. 3 shows ALT, AST and ALP levels in serum from mice of each group;
FIG. 4 is a graph showing the effect of ginsenoside CK on liver inflammatory factor levels;
FIG. 5 is the effect of ginsenoside CK on liver TLR4 mRNA levels;
FIG. 6 shows the effect of ginsenoside CK on IκBα phosphorylation.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
EXAMPLE 1 preparation of ginsenoside CK preparation
(1) Tablet: mixing ginsenoside CK 72g, dextrin 13g and microcrystalline cellulose 14g, spraying ethanol uniformly with a spray can to obtain powder, granulating, adding magnesium stearate 1g, mixing uniformly, and pressing.
(2) The granule comprises the following components: mixing ginsenoside CK 100g, dextrin 300g and sucrose 400g, spraying alcohol uniformly with spray can to obtain powder, granulating, and packaging into bags with 2g each.
(3) Soft capsule: taking 100g of ginsenoside CK, 600mL of water, 100mL of lecithin, 40050mL of PEG, 25mL of povidone and 10g of ascorbic acid, heating, stirring and mixing uniformly, and pouring the liquid medicine into a gelatin shell by a dripping method after the temperature is reduced to normal temperature to prepare the ginsenoside CK soft capsule.
(4) Oral liquid: taking 50g of ginsenoside CK, 20g of polysorbate, 2g of stevioside, 1g of sorbate, 5g of sodium carboxymethylcellulose and 20g of glycerin, adding purified water to 1L, stirring to uniformly mix, and filling into polyester bottles with 10mL of each bottle.
EXAMPLE 2 therapeutic Effect of ginsenoside CK on autoimmune hepatitis
(1) Establishment of autoimmune hepatitis model: after the ICR male mice are adaptively fed for one week, the weight of the ICR male mice is 18-20g, the ICR male mice are divided into a normal group, a model group and a low-dose and high-dose group of ginsenoside CK, 10 animals are in each group, normal group and model group are subjected to gastric lavage treatment every day, the mice are subjected to gastric lavage treatment according to doses of 20mg/kg and 40mg/kg respectively, conA is dissolved in normal saline after one week of administration, filtering and sterilizing are carried out, and other mice except the normal group are subjected to tail vein injection according to 15mg/kg ConA, and the normal group is injected with equal dose of normal saline. After 18 hours, mice were sacrificed and blood was collected and liver and spleen tissues were collected and saved for subsequent testing.
Results: within one week of administration of ginsenoside CK, the body weight of mice is not obviously reduced, which indicates that the ginsenoside CK has no obvious toxicity to the mice, and after ConA induction, the body weight of mice in a model group is obviously reduced, and the CK group is relatively relieved, which indicates that the CK can partially counteract the toxicity of ConA. In addition, the liver of the model group is dark red, the volume is reduced, the liver is in a damaged state, and the liver morphology in the CK group is recovered, which shows that the ginsenoside CK has a relieving effect on ConA-induced liver damage. Furthermore, the spleen of the model group is engorged with blood, has a volume expansion and a dark color, while the spleen of the CK group is in a normal form, which suggests that the ginsenoside CK has an effect of regulating the immune function. The results are shown in FIGS. 1, 2 and Table 1.
Table 1 weight and absolute liver and spleen masses of mice in each group
(2) Liver tissue H & E staining: the fixed liver was cut into tissue pieces of 10mm×3mm, then embedded and sectioned, and the sections were fixed on a glass slide, subjected to conventional H & E staining, observed under an optical microscope, and the pathological changes of the liver were recorded.
Results: the liver cells of the normal group are arranged tightly and orderly, the liver of the model group is obviously necrotized, the cell nuclei are contracted, the radial arrangement of the hepatic ropes is disappeared, the liver necrotic area of the mice of the CK low-dose group is obviously reduced, and the liver tissue morphology of the high-dose group is restored to be normal, which indicates that CK can inhibit ConA-induced liver tissue necrosis. The results are shown in FIG. 2.
(3) Detection of serum biochemical indexes: and centrifuging the collected blood, separating to obtain serum, analyzing a serum sample by adopting AST, ALT and ALP kits, and detecting the enzyme level in the serum.
Results: after ConA induction, the level of aminotransferase in serum rises sharply, which indicates that ConA causes serious damage to liver function, while the level of serum aminotransferase in CK pretreatment group is obviously reduced compared with model group, which indicates that CK has obvious relieving effect on liver function damage caused by ConA. The results are shown in FIG. 3.
(4) Expression of liver inflammatory factor: after freezing and grinding the collected liver tissue, adding normal saline and centrifuging, detecting the obtained supernatant by using a TNFa and IL-6ELISA kit, and analyzing the expression condition of several inflammatory factors in the liver.
Results: the data show that compared with the normal group, the liver inflammatory factor level of the model group treated by ConA alone is obviously increased, which indicates that the liver has serious inflammatory response, and the liver inflammatory factor expression level of the CK pretreatment group is returned, which indicates that the CK has an improvement effect on liver inflammation. The results are shown in FIG. 4.
(5) RT-qPCR (reverse transcription-quantitative polymerase chain reaction) detection of expression of inflammation-related genes and TLR4 in liver tissues: weighing 50mg of frozen liver tissue, adding a Trizol reagent to extract total RNA of the liver after freezing and grinding by liquid nitrogen, synthesizing cDNA by using a reverse transcription kit, amplifying by taking beta-action as an internal reference, reading a CT value after PCR reaction is finished, and calculating the expression condition of the TLR4 gene in each group.
Results: the mRNA level of TLR4 in the liver of ConA-induced model mice is obviously increased, and the expression level of TLR4 in a CK-treated group is reduced, which indicates that CK can inhibit the activity of TLR4 signals in the liver. The results are shown in FIG. 5.
(6) WB detection of phosphorylation of iκbα in liver tissue: weighing 50mg of frozen liver tissue, adding RIPA lysate for cracking, centrifuging, and taking supernatant to obtain total protein solution, wherein one part adopts BCA protein quantitative kit for total protein quantification, and the other part adopts the following steps: 1, adding a loading buffer solution in proportion, uniformly mixing, boiling for 5min, performing SDS-PAGE and membrane transfer, incubating an antibody, developing and recording the result.
Results: the level of IκBα phosphorylation was significantly up-regulated in the liver of ConA-induced model mice, while the expression level of p-IκBα was relatively reduced in the CK-treated group, indicating that CK can inhibit NF- κB signaling pathway activity by inhibiting IκBα phosphorylation. The results are shown in FIG. 6.
In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant points refer to the description of the method section.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (8)
1. Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis is provided.
2. The use according to claim 1, wherein the ginsenoside CK is the main or sole active ingredient in the medicament.
3. The use according to claim 1, wherein the immune hepatitis is concanavalin a-induced autoimmune hepatitis.
4. The use according to claim 1, wherein the ginsenoside CK is obtained by converting Rb1 extracted from ginseng, notoginseng or American ginseng by snailase, and the purity of the ginsenoside CK is not less than 95%.
5. The use according to claim 1, wherein the medicament is a composition of ginsenoside CK with pharmaceutical excipients and other auxiliary ingredients.
6. The use according to claim 1, wherein the medicament is an oral formulation, the dosage form comprising one of a tablet, a capsule, a granule, a suspension, a powder; wherein the tablet comprises a common tablet, a dispersible tablet, a chewable tablet or an enteric-coated tablet.
7. The use according to claim 1, wherein the single human dosage of ginsenoside CK is 10-100mg/kg.
8. The use according to claim 1, wherein the medicament is a medicament for ameliorating immune hepatitis through TLR4/NF- κb signaling.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311533828.9A CN117503782A (en) | 2023-11-17 | 2023-11-17 | Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311533828.9A CN117503782A (en) | 2023-11-17 | 2023-11-17 | Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117503782A true CN117503782A (en) | 2024-02-06 |
Family
ID=89754603
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311533828.9A Pending CN117503782A (en) | 2023-11-17 | 2023-11-17 | Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117503782A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116836818A (en) * | 2023-07-28 | 2023-10-03 | 陕西省微生物研究所 | Penicillium strain F8816 and application thereof |
-
2023
- 2023-11-17 CN CN202311533828.9A patent/CN117503782A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116836818A (en) * | 2023-07-28 | 2023-10-03 | 陕西省微生物研究所 | Penicillium strain F8816 and application thereof |
| CN116836818B (en) * | 2023-07-28 | 2024-03-22 | 陕西省微生物研究所 | Penicillium strain F8816 and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN117503782A (en) | Application of ginsenoside CK in preparing medicine for preventing and treating immune hepatitis | |
| CN115154481B (en) | A Chinese medicinal composition for preventing and treating atherosclerosis and coronary heart disease | |
| CN115364170B (en) | Sugar metabolism regulator and preparation method and application thereof | |
| CN114869928B (en) | Traditional Chinese medicine composition for treating cerebrovascular diseases and application thereof | |
| CN110123974A (en) | A kind of composition that treating hyperuricemia and preparation method | |
| CN100348228C (en) | New use of powder for regulating liver and spleen and its active part | |
| CN103784683B (en) | A kind of Chinese medicine composition treating obesity and its preparation method and application | |
| CN109223992B (en) | Traditional Chinese medicine composition for preventing and treating proteinuria and application thereof | |
| CN110613792B (en) | Traditional Chinese medicine composition with blood sugar reducing effect and preparation method and application thereof | |
| CN111297887B (en) | Preparation method and application of liver-protecting active component of Yunnan ginseng | |
| CN106822152B (en) | Pharmaceutical composition and application thereof | |
| CN111000852B (en) | Application of withanolide extract in physalis angulata in preparation of drugs for preventing or treating non-alcoholic fatty liver diseases | |
| CN118286310B (en) | Application of nux Prinsepiae or extract and composition thereof in treating and/or preventing liver injury | |
| CN107213323B (en) | Chinese medicinal compound preparation for nourishing yin, eliminating phlegm, resolving masses and detoxifying and application thereof | |
| CN114159516B (en) | Traditional Chinese medicine composition for preventing and treating hashimoto thyroiditis and application thereof | |
| CN116327826B (en) | A Chinese medicinal composition with antiinflammatory, analgesic or uric acid reducing effects and its preparation | |
| CN103565956B (en) | Traditional Chinese medicine composition for treating diabetes and preparation method thereof | |
| CN108815342B (en) | Traditional Chinese medicine composition for treating male infertility | |
| CN100446791C (en) | Chinese medicinal composition for treating hemorrhagic brain contusion and laceration | |
| CN1919277B (en) | Application of pharmaceutical composition in the process for preparing medicine of enhancing insulin sensitivity | |
| CN117982557A (en) | Use of fermented soya bean ginger or its extract and its composition | |
| CN118662553A (en) | Rhizoma anemones Altaicae or its extract, composition and application thereof | |
| CN116459311A (en) | Traditional Chinese medicine composition for treating type 2 diabetes and extract, preparation method and application thereof | |
| CN113769036A (en) | Traditional Chinese medicine compound composition for treating thyroid nodules and preparation method and application thereof | |
| CN117045739A (en) | Composition for treating type 2 diabetes and preparation method and application thereof |
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 |