JPWO2019077810A1 - Periostin and PKM2 secretagogue - Google Patents
Periostin and PKM2 secretagogue Download PDFInfo
- Publication number
- JPWO2019077810A1 JPWO2019077810A1 JP2019549111A JP2019549111A JPWO2019077810A1 JP WO2019077810 A1 JPWO2019077810 A1 JP WO2019077810A1 JP 2019549111 A JP2019549111 A JP 2019549111A JP 2019549111 A JP2019549111 A JP 2019549111A JP WO2019077810 A1 JPWO2019077810 A1 JP WO2019077810A1
- Authority
- JP
- Japan
- Prior art keywords
- agent
- muscle
- component
- improvement
- composition according
- 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.)
- Granted
Links
- 102100037765 Periostin Human genes 0.000 title claims abstract description 42
- 101710199268 Periostin Proteins 0.000 title claims abstract description 42
- 101001091538 Homo sapiens Pyruvate kinase PKM Proteins 0.000 title claims abstract 3
- 102100034911 Pyruvate kinase PKM Human genes 0.000 title claims abstract 3
- 230000000580 secretagogue effect Effects 0.000 title 1
- 239000000203 mixture Substances 0.000 claims abstract description 79
- 229930185474 acteoside Natural products 0.000 claims abstract description 51
- 210000000663 muscle cell Anatomy 0.000 claims abstract description 33
- 150000003839 salts Chemical class 0.000 claims abstract description 18
- 230000004663 cell proliferation Effects 0.000 claims abstract description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 77
- 230000006872 improvement Effects 0.000 claims description 45
- 210000003205 muscle Anatomy 0.000 claims description 44
- 210000003050 axon Anatomy 0.000 claims description 42
- QFRYQWYZSQDFOS-UHFFFAOYSA-N verbascoside Natural products CC1OC(COC2C(O)C(COC3OC(C(O)C(O)C3O)C(=O)O)OC(Oc4cc(O)cc5OC(=CC(=O)c45)c6ccc(O)c(O)c6)C2O)C(O)C(O)C1O QFRYQWYZSQDFOS-UHFFFAOYSA-N 0.000 claims description 41
- FBSKJMQYURKNSU-ZLSOWSIRSA-N acteoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@@H](CO)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O FBSKJMQYURKNSU-ZLSOWSIRSA-N 0.000 claims description 39
- 230000007659 motor function Effects 0.000 claims description 39
- FBSKJMQYURKNSU-UKQWSTALSA-N acteoside I Natural products C[C@@H]1O[C@H](O[C@@H]2[C@@H](O)[C@H](OCCc3ccc(O)c(O)c3)O[C@H](CO)[C@H]2OC(=O)C=Cc4ccc(O)c(O)c4)[C@H](O)[C@H](O)[C@H]1O FBSKJMQYURKNSU-UKQWSTALSA-N 0.000 claims description 37
- 235000013305 food Nutrition 0.000 claims description 36
- 238000011282 treatment Methods 0.000 claims description 36
- 230000001684 chronic effect Effects 0.000 claims description 35
- 208000020431 spinal cord injury Diseases 0.000 claims description 35
- 208000024891 symptom Diseases 0.000 claims description 24
- 201000000585 muscular atrophy Diseases 0.000 claims description 23
- 230000028327 secretion Effects 0.000 claims description 19
- 206010033799 Paralysis Diseases 0.000 claims description 17
- 206010028289 Muscle atrophy Diseases 0.000 claims description 16
- 208000029578 Muscle disease Diseases 0.000 claims description 16
- 230000020763 muscle atrophy Effects 0.000 claims description 16
- 208000012902 Nervous system disease Diseases 0.000 claims description 15
- 208000025966 Neurological disease Diseases 0.000 claims description 15
- 230000006870 function Effects 0.000 claims description 15
- 230000035755 proliferation Effects 0.000 claims description 15
- 208000001076 sarcopenia Diseases 0.000 claims description 14
- 230000003137 locomotive effect Effects 0.000 claims description 12
- 208000011580 syndromic disease Diseases 0.000 claims description 12
- 238000010255 intramuscular injection Methods 0.000 claims description 11
- 239000007927 intramuscular injection Substances 0.000 claims description 11
- 102000013009 Pyruvate Kinase Human genes 0.000 claims description 8
- 108020005115 Pyruvate Kinase Proteins 0.000 claims description 8
- 230000036541 health Effects 0.000 claims description 7
- 230000008439 repair process Effects 0.000 claims description 7
- 208000021642 Muscular disease Diseases 0.000 claims description 6
- 230000001737 promoting effect Effects 0.000 claims description 6
- 235000013402 health food Nutrition 0.000 claims description 5
- 210000000107 myocyte Anatomy 0.000 claims description 5
- 235000016709 nutrition Nutrition 0.000 claims description 5
- 238000000338 in vitro Methods 0.000 claims description 4
- 238000012423 maintenance Methods 0.000 claims description 4
- 239000013589 supplement Substances 0.000 claims description 4
- 241001125046 Sardina pilchardus Species 0.000 claims 1
- 235000019512 sardine Nutrition 0.000 claims 1
- 239000000284 extract Substances 0.000 description 44
- 241000699666 Mus <mouse, genus> Species 0.000 description 37
- 210000003141 lower extremity Anatomy 0.000 description 37
- 241000699670 Mus sp. Species 0.000 description 32
- -1 alkali metal salt Chemical class 0.000 description 30
- 238000000034 method Methods 0.000 description 28
- 201000010099 disease Diseases 0.000 description 26
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 26
- 210000000278 spinal cord Anatomy 0.000 description 22
- 239000002609 medium Substances 0.000 description 20
- 210000002363 skeletal muscle cell Anatomy 0.000 description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 18
- 210000002569 neuron Anatomy 0.000 description 18
- 208000027418 Wounds and injury Diseases 0.000 description 16
- 208000014674 injury Diseases 0.000 description 16
- 210000002027 skeletal muscle Anatomy 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 239000003814 drug Substances 0.000 description 14
- 230000006378 damage Effects 0.000 description 13
- 229940079593 drug Drugs 0.000 description 13
- 230000000694 effects Effects 0.000 description 13
- 241000196324 Embryophyta Species 0.000 description 12
- 241000336315 Cistanche salsa Species 0.000 description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- 210000003414 extremity Anatomy 0.000 description 11
- 239000002904 solvent Substances 0.000 description 10
- 230000007704 transition Effects 0.000 description 10
- 239000003636 conditioned culture medium Substances 0.000 description 9
- 238000001647 drug administration Methods 0.000 description 9
- 238000011156 evaluation Methods 0.000 description 9
- 239000000843 powder Substances 0.000 description 9
- 230000037396 body weight Effects 0.000 description 8
- 210000003314 quadriceps muscle Anatomy 0.000 description 7
- 238000007796 conventional method Methods 0.000 description 6
- 235000014113 dietary fatty acids Nutrition 0.000 description 6
- 239000000194 fatty acid Substances 0.000 description 6
- 229930195729 fatty acid Natural products 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 235000019198 oils Nutrition 0.000 description 6
- 239000008223 sterile water Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 229920002261 Corn starch Polymers 0.000 description 5
- 229930006000 Sucrose Natural products 0.000 description 5
- 239000008120 corn starch Substances 0.000 description 5
- 229940099112 cornstarch Drugs 0.000 description 5
- 235000014134 echinacea Nutrition 0.000 description 5
- NJYVDFDTLLZVMG-UHFFFAOYSA-N echinacoside Natural products CC1OC(OC2C(O)C(OCCc3ccc(O)c(O)c3)OC(COC4OC(CO)C(O)C(O)C4O)C2OC(=O)C=Cc5cc(O)cc(O)c5)C(O)C(O)C1O NJYVDFDTLLZVMG-UHFFFAOYSA-N 0.000 description 5
- FSBUXLDOLNLABB-ISAKITKMSA-N echinacoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O FSBUXLDOLNLABB-ISAKITKMSA-N 0.000 description 5
- 235000013373 food additive Nutrition 0.000 description 5
- 239000002778 food additive Substances 0.000 description 5
- 230000003387 muscular Effects 0.000 description 5
- 210000005036 nerve Anatomy 0.000 description 5
- 239000000546 pharmaceutical excipient Substances 0.000 description 5
- 239000005720 sucrose Substances 0.000 description 5
- 238000001356 surgical procedure Methods 0.000 description 5
- 239000003826 tablet Substances 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 235000013361 beverage Nutrition 0.000 description 4
- 229920002678 cellulose Polymers 0.000 description 4
- 239000001913 cellulose Substances 0.000 description 4
- 235000010980 cellulose Nutrition 0.000 description 4
- 210000003710 cerebral cortex Anatomy 0.000 description 4
- 235000009508 confectionery Nutrition 0.000 description 4
- 239000012228 culture supernatant Substances 0.000 description 4
- 201000006938 muscular dystrophy Diseases 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 210000000130 stem cell Anatomy 0.000 description 4
- 239000006188 syrup Substances 0.000 description 4
- 235000020357 syrup Nutrition 0.000 description 4
- 241000283707 Capra Species 0.000 description 3
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 3
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 3
- 244000133098 Echinacea angustifolia Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000012981 Hank's balanced salt solution Substances 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 241001508464 Orobanche Species 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 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 3
- 239000011248 coating agent Substances 0.000 description 3
- 239000006071 cream Substances 0.000 description 3
- 239000007884 disintegrant Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- 235000019197 fats Nutrition 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 235000013376 functional food Nutrition 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 235000015110 jellies Nutrition 0.000 description 3
- 230000001114 myogenic effect Effects 0.000 description 3
- 210000002241 neurite Anatomy 0.000 description 3
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 3
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 3
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000002062 proliferating effect Effects 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- 239000013049 sediment Substances 0.000 description 3
- 235000012239 silicon dioxide Nutrition 0.000 description 3
- 239000007901 soft capsule Substances 0.000 description 3
- 239000000600 sorbitol Substances 0.000 description 3
- 235000010356 sorbitol Nutrition 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- 238000002054 transplantation Methods 0.000 description 3
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 208000017667 Chronic Disease Diseases 0.000 description 2
- 208000034656 Contusions Diseases 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 2
- 108010044467 Isoenzymes Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 229920000881 Modified starch Polymers 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 229920002125 Sokalan® Polymers 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 244000299461 Theobroma cacao Species 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 150000001298 alcohols Chemical class 0.000 description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 235000001465 calcium Nutrition 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 230000009519 contusion Effects 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 235000015872 dietary supplement Nutrition 0.000 description 2
- 235000021186 dishes Nutrition 0.000 description 2
- NOPFSRXAKWQILS-UHFFFAOYSA-N docosan-1-ol Chemical compound CCCCCCCCCCCCCCCCCCCCCCO NOPFSRXAKWQILS-UHFFFAOYSA-N 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 2
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 2
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 2
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 2
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 2
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 2
- 238000012744 immunostaining Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 238000001361 intraarterial administration Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000314 lubricant Substances 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
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 229920000609 methyl cellulose Polymers 0.000 description 2
- 239000001923 methylcellulose Substances 0.000 description 2
- 235000010981 methylcellulose Nutrition 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- 210000002161 motor neuron Anatomy 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 235000012149 noodles Nutrition 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 239000006191 orally-disintegrating tablet Substances 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 230000000737 periodic effect Effects 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000003518 presynaptic effect Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 235000015067 sauces Nutrition 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000008159 sesame oil Substances 0.000 description 2
- 235000011803 sesame oil Nutrition 0.000 description 2
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 2
- 235000014214 soft drink Nutrition 0.000 description 2
- 239000003549 soybean oil Substances 0.000 description 2
- 235000012424 soybean oil Nutrition 0.000 description 2
- 208000002320 spinal muscular atrophy Diseases 0.000 description 2
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N squalane Chemical compound CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- ZMANZCXQSJIPKH-UHFFFAOYSA-N triethylamine Natural products CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- KDSWDGKIENPKLB-QJDQKFITSA-N verbascoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)CCC=2C=C(O)C(O)=CC=2)[C@@H](CO)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O KDSWDGKIENPKLB-QJDQKFITSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- NOOLISFMXDJSKH-UTLUCORTSA-N (+)-Neomenthol Chemical compound CC(C)[C@@H]1CC[C@@H](C)C[C@@H]1O NOOLISFMXDJSKH-UTLUCORTSA-N 0.000 description 1
- BGRXBNZMPMGLQI-UHFFFAOYSA-N 2-octyldodecyl tetradecanoate Chemical compound CCCCCCCCCCCCCC(=O)OCC(CCCCCCCC)CCCCCCCCCC BGRXBNZMPMGLQI-UHFFFAOYSA-N 0.000 description 1
- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical compound O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 239000012103 Alexa Fluor 488 Substances 0.000 description 1
- 239000012110 Alexa Fluor 594 Substances 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 208000002109 Argyria Diseases 0.000 description 1
- 241000416162 Astragalus gummifer Species 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 239000012583 B-27 Supplement Substances 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 108010058699 Choline O-acetyltransferase Proteins 0.000 description 1
- 102100023460 Choline O-acetyltransferase Human genes 0.000 description 1
- 102000005598 Chondroitin Sulfate Proteoglycans Human genes 0.000 description 1
- 108010059480 Chondroitin Sulfate Proteoglycans Proteins 0.000 description 1
- 244000223760 Cinnamomum zeylanicum Species 0.000 description 1
- 241000336291 Cistanche deserticola Species 0.000 description 1
- 241000336316 Cistanche tubulosa Species 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 240000006766 Cornus mas Species 0.000 description 1
- 235000003363 Cornus mas Nutrition 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 229920002785 Croscarmellose sodium Polymers 0.000 description 1
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 description 1
- 238000000116 DAPI staining Methods 0.000 description 1
- NOOLISFMXDJSKH-UHFFFAOYSA-N DL-menthol Natural products CC(C)C1CCC(C)CC1O NOOLISFMXDJSKH-UHFFFAOYSA-N 0.000 description 1
- 102000007260 Deoxyribonuclease I Human genes 0.000 description 1
- 108010008532 Deoxyribonuclease I Proteins 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 238000001061 Dunnett's test Methods 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 241000521834 Echinacea pallida Species 0.000 description 1
- 240000004530 Echinacea purpurea Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 240000008620 Fagopyrum esculentum Species 0.000 description 1
- 235000009419 Fagopyrum esculentum Nutrition 0.000 description 1
- 206010018341 Gliosis Diseases 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000979001 Homo sapiens Methionine aminopeptidase 2 Proteins 0.000 description 1
- 101000969087 Homo sapiens Microtubule-associated protein 2 Proteins 0.000 description 1
- 239000004354 Hydroxyethyl cellulose Substances 0.000 description 1
- 229920000663 Hydroxyethyl cellulose Polymers 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 201000010743 Lambert-Eaton myasthenic syndrome Diseases 0.000 description 1
- 240000005856 Lyophyllum decastes Species 0.000 description 1
- 235000013194 Lyophyllum decastes Nutrition 0.000 description 1
- 102100023174 Methionine aminopeptidase 2 Human genes 0.000 description 1
- 206010058799 Mitochondrial encephalomyopathy Diseases 0.000 description 1
- 208000002740 Muscle Rigidity Diseases 0.000 description 1
- 201000009623 Myopathy Diseases 0.000 description 1
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 1
- 108010025020 Nerve Growth Factor Proteins 0.000 description 1
- 102000007072 Nerve Growth Factors Human genes 0.000 description 1
- 241000207836 Olea <angiosperm> Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 244000239204 Plantago lanceolata Species 0.000 description 1
- 235000010503 Plantago lanceolata Nutrition 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- 229920001800 Shellac Polymers 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 208000026214 Skeletal muscle atrophy Diseases 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 102000004874 Synaptophysin Human genes 0.000 description 1
- 108090001076 Synaptophysin Proteins 0.000 description 1
- 235000009470 Theobroma cacao Nutrition 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- YKTSYUJCYHOUJP-UHFFFAOYSA-N [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] Chemical compound [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] YKTSYUJCYHOUJP-UHFFFAOYSA-N 0.000 description 1
- 229940124532 absorption promoter Drugs 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 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
- 230000001154 acute effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 229940009868 aluminum magnesium silicate Drugs 0.000 description 1
- WMGSQTMJHBYJMQ-UHFFFAOYSA-N aluminum;magnesium;silicate Chemical compound [Mg+2].[Al+3].[O-][Si]([O-])([O-])[O-] WMGSQTMJHBYJMQ-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 238000002266 amputation Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000037444 atrophy Effects 0.000 description 1
- 230000003376 axonal effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 235000015895 biscuits Nutrition 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 239000006189 buccal tablet Substances 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 description 1
- 239000001354 calcium citrate Substances 0.000 description 1
- 229960004256 calcium citrate Drugs 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000012054 celltiter-glo Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000007910 chewable tablet Substances 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 235000017803 cinnamon Nutrition 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000003240 coconut oil Substances 0.000 description 1
- 235000019864 coconut oil Nutrition 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 235000014510 cooky Nutrition 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 210000003618 cortical neuron Anatomy 0.000 description 1
- 229960001681 croscarmellose sodium Drugs 0.000 description 1
- 229960000913 crospovidone Drugs 0.000 description 1
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 235000021438 curry Nutrition 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 210000001787 dendrite Anatomy 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- SZXQTJUDPRGNJN-UHFFFAOYSA-N dipropylene glycol Chemical compound OCCCOCCCO SZXQTJUDPRGNJN-UHFFFAOYSA-N 0.000 description 1
- 108010007093 dispase Proteins 0.000 description 1
- 229960000735 docosanol Drugs 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 239000003221 ear drop Substances 0.000 description 1
- 229940047652 ear drops Drugs 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000008393 encapsulating agent Substances 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000010696 ester oil Substances 0.000 description 1
- 150000002169 ethanolamines Chemical class 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 239000003885 eye ointment Substances 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 229910021482 group 13 metal Inorganic materials 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000007902 hard capsule Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 235000019447 hydroxyethyl cellulose Nutrition 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 235000008446 instant noodles Nutrition 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000007914 intraventricular administration Methods 0.000 description 1
- 235000015094 jam Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 239000000944 linseed oil Substances 0.000 description 1
- 235000021388 linseed oil Nutrition 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 229910003002 lithium salt Inorganic materials 0.000 description 1
- 159000000002 lithium salts Chemical class 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 229940031703 low substituted hydroxypropyl cellulose Drugs 0.000 description 1
- 210000005014 lower spinal cord Anatomy 0.000 description 1
- 210000005230 lumbar spinal cord Anatomy 0.000 description 1
- 210000004705 lumbosacral region Anatomy 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 239000000391 magnesium silicate Substances 0.000 description 1
- 229910052919 magnesium silicate Inorganic materials 0.000 description 1
- 235000019792 magnesium silicate Nutrition 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000003264 margarine Substances 0.000 description 1
- 235000013310 margarine Nutrition 0.000 description 1
- 235000010746 mayonnaise Nutrition 0.000 description 1
- 239000008268 mayonnaise Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000001525 mentha piperita l. herb oil Substances 0.000 description 1
- 229940041616 menthol Drugs 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000009525 mild injury Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- JXTPJDDICSTXJX-UHFFFAOYSA-N n-Triacontane Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC JXTPJDDICSTXJX-UHFFFAOYSA-N 0.000 description 1
- 239000007923 nasal drop Substances 0.000 description 1
- 229940100662 nasal drops Drugs 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000001178 neural stem cell Anatomy 0.000 description 1
- 230000001272 neurogenic effect Effects 0.000 description 1
- 210000000715 neuromuscular junction Anatomy 0.000 description 1
- 201000001119 neuropathy Diseases 0.000 description 1
- 230000007823 neuropathy Effects 0.000 description 1
- 239000003900 neurotrophic factor Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229940073665 octyldodecyl myristate Drugs 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 208000021090 palsy Diseases 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 235000019477 peppermint oil Nutrition 0.000 description 1
- 208000033808 peripheral neuropathy Diseases 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000004584 polyacrylic acid Substances 0.000 description 1
- 229920002503 polyoxyethylene-polyoxypropylene Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 235000019422 polyvinyl alcohol Nutrition 0.000 description 1
- 235000013809 polyvinylpolypyrrolidone Nutrition 0.000 description 1
- 229920000523 polyvinylpolypyrrolidone Polymers 0.000 description 1
- 235000021395 porridge Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 235000013606 potato chips Nutrition 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 229940088417 precipitated calcium carbonate Drugs 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 235000020991 processed meat Nutrition 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- JUJWROOIHBZHMG-UHFFFAOYSA-N pyridine Substances C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000035807 sensation Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 229920002545 silicone oil Polymers 0.000 description 1
- 230000025185 skeletal muscle atrophy Effects 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 229940032094 squalane Drugs 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 235000013547 stew Nutrition 0.000 description 1
- 239000006190 sub-lingual tablet Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000012353 t test Methods 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 125000005270 trialkylamine group Chemical group 0.000 description 1
- 235000013337 tricalcium citrate Nutrition 0.000 description 1
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical class CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 1
- 210000001364 upper extremity Anatomy 0.000 description 1
- 239000006216 vaginal suppository Substances 0.000 description 1
- 239000000003 vaginal tablet Substances 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229920003169 water-soluble polymer Polymers 0.000 description 1
- 229940100445 wheat starch Drugs 0.000 description 1
- 239000008256 whipped cream Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/64—Orobanchaceae (Broom-rape family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/45—Transferases (2)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Neurology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medical Informatics (AREA)
- Nutrition Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Neurosurgery (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Microbiology (AREA)
- Physical Education & Sports Medicine (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
筋細胞の増殖等に有用な剤又は組成物を提供する。剤又は組成物を、アクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)やペリオスチン及びPKM2から選択された少なくとも1種の成分(B)で構成する。Provided is an agent or composition useful for muscle cell proliferation and the like. The agent or composition is composed of at least one component (A) selected from acteosides, echinacosides and salts thereof, and at least one component (B) selected from periostin and PKM2.
Description
本発明は、ペリオスチンやPKM2の分泌(発現)を促進(増大)しうる剤又は組成物等に関する。 The present invention relates to an agent or composition capable of promoting (increasing) the secretion (expression) of periostin and PKM2.
脊髄損傷では、外傷性に挫滅あるいは離断した脊髄内で軸索が断裂し、損傷脊髄部位およびその下位脊髄が支配する体幹・上下肢の運動と感覚が機能不全に陥る。 In spinal cord injury, axons rupture in the traumatically crushed or transected spinal cord, resulting in dysfunction of the injured spinal cord and the movement and sensation of the trunk and upper and lower limbs controlled by the lower spinal cord.
脊髄損傷に対する臨床での対処法は、受傷直後のステロイド剤の大量投与であったが、機能回復への実効性は疑問視されており(非特許文献1)、副作用のリスクが大きいため推奨されていないのが現状である。比較的損傷が軽度な場合は、リハビリテーションによる残存機能の強化で機能改善が見られる場合もあるが、受傷後数か月―数年の時期を経た“慢性期”の患者に対しては、機能回復の可能性は極めて低い。 The clinical treatment for spinal cord injury was to administer a large amount of steroids immediately after the injury, but its effectiveness for functional recovery has been questioned (Non-Patent Document 1), and it is recommended because of the high risk of side effects. The current situation is that it has not. In relatively mild injuries, rehabilitation may improve residual function, but in "chronic" patients months-years after injury, function The chances of recovery are extremely low.
近年、神経幹細胞やiPS細胞を応用した再生医療が次世代の脊髄損傷の治療戦略として有望視され、精力的に基礎研究が進められているが(非特許文献2及び3)、マウスでの神経幹/前駆細胞の移植では、受傷後9日後(亜急性期)までの移植では損傷部での軸索伸展および運動機能の回復が認められるのに対し、受傷後42日後(慢性期)の移植では全く無効である(非特許文献4)。 In recent years, regenerative medicine applying neural stem cells and iPS cells has been seen as a promising next-generation treatment strategy for spinal cord injury, and basic research is being vigorously pursued (Non-Patent Documents 2 and 3), but nerves in mice. In stem / progenitor cell transplantation, axillary extension and recovery of motor function are observed in the injured area up to 9 days after injury (subacute phase), whereas transplantation 42 days after injury (chronic phase) Is completely invalid (Non-Patent Document 4).
慢性期脊髄損傷動物に対して、神経幹/前駆細胞の移植と神経栄養因子の同時処置(非特許文献5)、あるいはChABC*の同時処置(*グリア性瘢痕から分泌される軸索阻害因子のコンドロイチン硫酸プロテオグリカンを分解する酵素)(非特許文献6)、あるいはトレッドミル歩行による運動負荷の同時処置(非特許文献7)が試されているが、機能回復の程度は未だ限定的である。つまり、受傷後慢性期の脊髄損傷の機能回復は依然として極めて困難である。 Transplantation of nerve stem / progenitor cells and simultaneous treatment of neurotrophic factors (Non-Patent Document 5) or simultaneous treatment of ChABC * (* axon inhibitor secreted from glial scars) for animals with SCI An enzyme that decomposes chondroitin sulfate proteoglycan) (Non-Patent Document 6) or simultaneous treatment of exercise load by walking with a tread mill (Non-Patent Document 7) has been tried, but the degree of functional recovery is still limited. In other words, it is still extremely difficult to recover the function of spinal cord injury in the chronic phase after injury.
本発明の目的は、ペリオスチンやPKM2(ピルビン酸キナーゼM2、ピルビン酸キナーゼM2アイソザイム)の分泌を促進しうる剤又は組成物等を提供することにある。 An object of the present invention is to provide an agent or composition capable of promoting the secretion of periostin or PKM2 (pyruvate kinase M2, pyruvate kinase M2 isozyme).
本発明の他の目的は、軸索を伸展(伸長)させたり、筋細胞の増殖や筋萎縮を改善(向上)しうる剤又は組成物等を提供することにある。 Another object of the present invention is to provide an agent or composition capable of extending (elongating) axons and improving (improving) myocyte proliferation and muscular atrophy.
本発明の他の目的は、種々の症状・疾患の改善や、運動機能や体力の改善(向上)・維持に有用な剤又は組成物等を提供することにある。 Another object of the present invention is to provide an agent or composition useful for improving various symptoms / diseases and improving (improving) / maintaining motor function and physical strength.
本発明者らは、鋭意検討を重ねた結果、特定成分(アクテオシドやエキナコシド)が、ペリオスチンやPKM2の分泌を促進すること、また、意外なことに、このようなペリオスチンやPKM2の分泌を介して、全身性にシグナルが拡散され、特に、筋細胞が増殖されたり、筋肉量が増加したり、軸索(神経突起)の伸展(伸長)が促進されること等を見出し、本発明を完成した。 As a result of diligent studies, the present inventors have found that specific components (acteosides and echinacosides) promote the secretion of periostin and PKM2, and surprisingly, through the secretion of such periostin and PKM2. The present invention was completed by finding that signals are diffused systemically, muscle cells are proliferated, muscle mass is increased, and axon (neurite) extension (extension) is promoted. ..
すなわち、本発明は以下の発明等に関する。
[1]
アクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)を含み、ペリオスチン及び/又はピルビン酸キナーゼM2の分泌促進のための剤又は組成物。
[2]
アクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)を含み、筋細胞の増殖、筋肉(又は筋肉量、以下同じ)の増加及び/又は筋萎縮の改善(治療)のための剤又は組成物。
[3]
さらに、軸索を伸展及び/又は修復させるための、[2]記載の剤又は組成物。
[4]
ペリオスチン及びPKM2から選択された少なくとも1種の成分(B)[及びアクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)]を含み、軸索の伸展、軸索の修復、筋細胞の増殖、筋肉の増加、及び/又は筋萎縮の改善(治療)のための剤又は組成物。
[5]
成分(B)が生体外に存在するものである[4]記載の剤又は組成物。
[6]
下記の(1)〜(6)から選択された少なくとも1つの用途に使用するための、[1]〜[5]のいずれかに記載の剤又は組成物。
(1)神経疾患の治療及び/又は改善
(2)筋疾患の治療及び/又は改善
(3)サルコペニアの治療及び/又は改善
(4)ロコモーティブシンドロームの治療及び/又は改善
(5)麻痺の治療及び/又は改善
(6)運動機能及び/又は体力の改善(向上)及び/又は維持
[7]
神経疾患、筋疾患、サルコペニア、ロコモーティブシンドローム及び麻痺が、慢性期の症状である[6]記載の剤又は組成物。
[8]
慢性期の脊髄損傷を治療及び/又は改善するための[1]〜[7]のいずれかに記載の剤又は組成物。
[9]
成分(A)として、少なくともアクテオシドを含む[1]〜[3]及び[6]〜[8]のいずれかに記載の剤又は組成物。
[10]
成分(A)を含有する成分として、ニクジュウヨウ成分を含有する[1]〜[3]及び[6]〜[9]のいずれかに記載の剤又は組成物。
[11]
筋肉注射又は経口用である[1]〜[10]のいずれかに記載の剤又は組成物。
[12]
飲食品である[1]〜[11]のいずれかに記載の剤又は組成物。
[13]
サプリメント、健康食品、機能性表示食品、栄養機能食品又は特定保健用食品である[1]〜[12]のいずれかに記載の剤又は組成物。
[14]
アクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)[又は成分(A)を含む剤又は組成物]を用いて、ペリオスチン及び/又はピルビン酸キナーゼM2の分泌を促進させる方法。
[15]
アクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)[又は成分(A)を含む剤又は組成物]を用いて、筋細胞を増殖させる、筋肉を増加(増大)させる及び/又は筋萎縮を改善(治療)する方法。
[16]
さらに、軸索を伸展及び/又は修復させる、[15]記載の方法。
[17]
ペリオスチン及びPKM2から選択された少なくとも1種の成分(B)[及びアクテオシド、エキナコシド及びこれらの塩から選択された少なくとも1種の成分(A)を含む剤又は組成物]を用いて、軸索を伸展、軸索を修復、筋細胞を増殖、筋肉を増加、及び/又は筋萎縮を改善(治療)させる方法。
[18]
成分(B)が生体外に存在するものである[17]記載の方法。
[19]
下記の(1)〜(6)から選択された少なくとも1つを含む、[14]〜[18]のいずれかに記載の方法。
(1)神経疾患を治療及び/又は改善する
(2)筋疾患を治療及び/又は改善する
(3)サルコペニアを治療及び/又は改善する
(4)ロコモーティブシンドロームを治療及び/又は改善する
(5)麻痺を治療及び/又は改善する
(6)運動機能及び/又は体力を改善(向上)及び/又は維持する
[20]
神経疾患、筋疾患、サルコペニア、ロコモーティブシンドローム及び麻痺が、慢性期の症状である[19]記載の方法。
[21]
慢性期の脊髄損傷を治療及び/又は改善する、[14]〜[20]のいずれかに記載の方法。
[22]
成分(A)として、少なくともアクテオシドを含む[14]〜[21]のいずれかに記載の方法。
[23]
成分(A)を含有する成分として、ニクジュウヨウ成分を含有する[14]〜[22]のいずれかに記載の方法。
[24]
筋肉注射又は経口投与する、[14]〜[23]のいずれかに記載の方法。
[25]
剤又は組成物が飲食品である[14]〜[24]のいずれかに記載の方法。
[26]
剤又は組成物が、サプリメント、健康食品、機能性表示食品、栄養機能食品又は特定保健用食品である[14]〜[25]のいずれかに記載の方法。That is, the present invention relates to the following inventions and the like.
[1]
An agent or composition containing at least one component (A) selected from acteosides, echinacosides and salts thereof, and for promoting the secretion of periostin and / or pyruvate kinase M2.
[2]
Contains at least one component (A) selected from acteosides, echinacosides and salts thereof, for the growth of muscle cells, the increase of muscle (or muscle mass, the same applies hereinafter) and / or the improvement (treatment) of muscle atrophy. Agent or composition.
[3]
Further, the agent or composition according to [2] for extending and / or repairing an axon.
[4]
Containing at least one component (B) [and at least one component (A) selected from acteosides, echinacoside and salts thereof] selected from periostin and PKM2, axon extension, axon repair, An agent or composition for improving (treating) muscle cell proliferation, muscle gain, and / or muscle atrophy.
[5]
The agent or composition according to [4], wherein the component (B) is present in vitro.
[6]
The agent or composition according to any one of [1] to [5] for use in at least one application selected from the following (1) to (6).
(1) Treatment and / or improvement of neurological disorders (2) Treatment and / or improvement of muscle disorders (3) Treatment and / or improvement of sarcopenia (4) Treatment and / or improvement of locomotive syndrome (5) Treatment and improvement of paralysis / Or improvement (6) Improvement (improvement) and / or maintenance of motor function and / or physical strength [7]
The agent or composition according to [6], wherein neurological disease, muscle disease, sarcopenia, locomotive syndrome and paralysis are symptoms of the chronic phase.
[8]
The agent or composition according to any one of [1] to [7] for treating and / or ameliorating spinal cord injury in the chronic phase.
[9]
The agent or composition according to any one of [1] to [3] and [6] to [8], which contains at least an acteoside as the component (A).
[10]
The agent or composition according to any one of [1] to [3] and [6] to [9], which contains a syrup component as a component containing the component (A).
[11]
The agent or composition according to any one of [1] to [10] for intramuscular injection or oral use.
[12]
The agent or composition according to any one of [1] to [11], which is a food or drink.
[13]
The agent or composition according to any one of [1] to [12], which is a supplement, a health food, a food with a functional claim, a food with a nutritional function, or a food for specified health use.
[14]
A method for promoting the secretion of periostin and / or pyruvate kinase M2 by using at least one component (A) [or an agent or composition containing the component (A)] selected from acteosides, echinacosides and salts thereof. ..
[15]
Using at least one component (A) [or an agent or composition containing component (A)] selected from acteosides, echinacosides and salts thereof, muscle cells are proliferated, muscles are increased (increased) and / Or a method of improving (treating) muscle atrophy.
[16]
[15] The method of [15], wherein the axon is extended and / or repaired.
[17]
Axons are squeezed using at least one component (B) selected from periostin and PKM2 [and an agent or composition containing at least one component (A) selected from acteosides, echinacoside and salts thereof]. A method of extending, repairing axons, proliferating muscle cells, gaining muscle, and / or improving (treating) muscle atrophy.
[18]
The method according to [17], wherein the component (B) is present outside the living body.
[19]
The method according to any one of [14] to [18], which comprises at least one selected from the following (1) to (6).
(1) Treat and / or improve neurological disorders (2) Treat and / or improve muscle disorders (3) Treat and / or improve sarcopenia (4) Treat and / or improve locomotive syndrome (5) Treat and / or improve paralysis (6) Improve (improve) and / or maintain motor function and / or physical fitness [20]
The method according to [19], wherein neurological disease, muscle disease, sarcopenia, locomotive syndrome and paralysis are symptoms of the chronic phase.
[21]
The method according to any of [14] to [20], which treats and / or improves spinal cord injury in the chronic phase.
[22]
The method according to any one of [14] to [21], which comprises at least an acteoside as the component (A).
[23]
The method according to any one of [14] to [22], which contains a Nikujuyo component as a component containing the component (A).
[24]
The method according to any one of [14] to [23], which is administered intramuscularly or orally.
[25]
The method according to any one of [14] to [24], wherein the agent or composition is a food or drink.
[26]
The method according to any one of [14] to [25], wherein the agent or composition is a supplement, a health food, a food with a functional claim, a food with a nutritional function, or a food for specified health use.
本発明では、ペリオスチンやPKM2の分泌を促進しうる。そして、本発明者らは、ペリオスチンやPKM2が、筋細胞に対しては増殖作用、神経細胞に対しては軸索伸展作用を有することを確認している。そのため、これらの機能に関連する種々の疾患・症状等の治療及び/又は改善(向上)に有用である。 In the present invention, the secretion of periostin and PKM2 can be promoted. Then, the present inventors have confirmed that periostin and PKM2 have a proliferative action on muscle cells and an axon extension action on nerve cells. Therefore, it is useful for the treatment and / or improvement (improvement) of various diseases / symptoms related to these functions.
本発明の他の態様では、筋細胞の増殖(又は筋細胞の増殖の促進又は筋肉量の増加)や筋萎縮を改善(向上)しうる。 In another aspect of the present invention, muscle cell proliferation (or promotion of muscle cell proliferation or increase in muscle mass) and muscle atrophy can be improved (improved).
そのため、本発明は、筋細胞の低下や筋萎縮などに起因又は関連する疾患・症状[例えば、神経疾患(脊髄損傷など)、筋疾患、サルコペニア、ロコモーティブシンドローム、麻痺(例えば、手及び/又は足の麻痺、四肢の麻痺)など、特に慢性期におけるこれらの疾患・症状]などの治療ないし改善のために好適に適用しうる。 Therefore, the present invention relates to diseases / symptoms caused or related to muscle cell depletion, muscle atrophy, etc. [for example, neurological disease (spinal cord injury, etc.), muscle disease, sarcopenia, locomotive syndrome, paralysis (eg, hand and / or foot). It can be suitably applied for the treatment or improvement of these diseases / symptoms, such as paralysis of limbs, paralysis of limbs), especially in the chronic phase.
また、筋細胞の増殖を促すため、何らかの症状・疾患に至っているか否かを問わず、運動機能や体力を改善ないし維持しうる。 In addition, in order to promote the proliferation of muscle cells, it is possible to improve or maintain motor function and physical strength regardless of whether or not any symptom or disease has occurred.
なお、このような筋細胞の増殖等は、ペリオスチンやPKM2(ピルビン酸キナーゼM2)の分泌促進を介して、行われてもよい。 In addition, such proliferation of muscle cells may be carried out through promotion of secretion of periostin and PKM2 (pyruvate kinase M2).
<剤・組成物>
本発明の剤又は組成物は、特定の成分(A)[及び/又は成分(B)]を含み、種々の用途において使用可能である。<Agent / Composition>
The agent or composition of the present invention contains a specific component (A) [and / or component (B)] and can be used in various applications.
[用途]
本発明の剤又は組成物(第1の態様の剤又は組成物)は、ペリオスチン及び/又はピルビン酸キナーゼM2(PKM2、ピルビン酸キナーゼM2アイソザイム)の分泌促進に有用である。なお、ペリオスチン及び/又はPKM2は、特に限定されないが、特に、筋肉(骨格筋など)又は筋細胞から分泌させてもよい。[Use]
The agent or composition of the present invention (the agent or composition of the first aspect) is useful for promoting the secretion of periostin and / or pyruvate kinase M2 (PKM2, pyruvate kinase M2 isozyme). The periostin and / or PKM2 is not particularly limited, but may be secreted from muscles (skeletal muscles and the like) or muscle cells.
そして、本発明者らの検討によれば、ペリオスチンやPKM2の分泌により(分泌を介して)、筋細胞が増殖すること、筋肉量が増加することや、軸索が伸展又は修復することを確認している。 Then, according to the study by the present inventors, it was confirmed that the secretion of periostin and PKM2 (via secretion) causes the proliferation of muscle cells, the increase in muscle mass, and the extension or repair of axons. doing.
そのため、第1の態様の剤又は組成物は、後述のように、筋細胞の増殖、筋肉量の増加(増大)や軸索の伸展に関与する症状・疾患等の改善・向上に適用しうる。 Therefore, the agent or composition of the first aspect can be applied to the improvement / improvement of symptoms / diseases related to muscle cell proliferation, muscle mass increase (increase) and axon extension, as described later. ..
第2の態様の剤又は組成物は、筋細胞の増殖(増殖の促進)、筋肉量の増加及び/又は筋萎縮の改善(治療)のために有用である。 The agent or composition of the second aspect is useful for myocyte proliferation (promotion of proliferation), increase in muscle mass and / or improvement of muscle atrophy (treatment).
筋細胞や筋萎縮において、対象となる筋肉は、特に限定されないが、特に骨格筋、中でも、運動機能に直結しやすい骨格筋、例えば、手や足の筋肉を少なくとも含んでいてもよい。 In muscle cells and muscular atrophy, the target muscles are not particularly limited, but may include at least skeletal muscles, particularly skeletal muscles that are easily directly linked to motor function, for example, muscles of hands and feet.
このような第2の態様の剤又は組成物は、軸索の伸展及び/又は修復機能を有していてもよい。具体的には、第2の態様の剤又は組成物は、神経突起[特に萎縮又は欠損した神経突起(軸索や樹状突起)]の修復や伸展(伸長)のために用いてもよい。 The agent or composition of such a second aspect may have an axon extension and / or repair function. Specifically, the agent or composition of the second aspect may be used for repair or extension (elongation) of neurites [particularly atrophied or defective neurites (axons and dendrites)].
第2の態様の剤又は組成物は、上記のように、筋細胞の増殖機能、筋肉量の増大機能、や筋萎縮の改善機能(さらには、軸索の伸展・修復機能)を有しているため、このような機能に関連して、種々の用途に適用可能である。 As described above, the agent or composition of the second aspect has a muscle cell proliferation function, a muscle mass increasing function, and a muscular atrophy improving function (further, an axon extension / repair function). Therefore, it can be applied to various uses in relation to such a function.
例えば、第2の態様の剤又は組成物は、下記の(1)〜(5)から選択された少なくとも1つの用途に使用してもよい。 For example, the agent or composition of the second aspect may be used for at least one application selected from the following (1) to (5).
(1)神経疾患の治療及び/又は改善
(2)筋疾患の治療及び/又は改善
(3)サルコペニアの治療及び/又は改善
(4)ロコモーティブシンドロームの治療及び/又は改善
(5)麻痺(手及び/又は足の麻痺、四肢の麻痺など)の治療及び/又は改善(1) Treatment and / or improvement of neurological disorders (2) Treatment and / or improvement of muscle disorders (3) Treatment and / or improvement of sarcopenia (4) Treatment and / or improvement of locomotive syndrome (5) Paralysis (hands and hands and / Or treatment and / or improvement of foot paralysis, limb paralysis, etc.)
神経疾患としては、例えば、脊髄損傷、筋萎縮性脊索硬化症(ALS)などが挙げられる。 Neurological disorders include, for example, spinal cord injury, amyotrophic lateral sclerosis (ALS), and the like.
筋疾患は、筋原性疾患(ミオパチー)、神経原性疾患(ニューロパチー)などであってもよく、特に、筋原性疾患であってもよい。 The myopathic disease may be a myopathic disease (myopathy), a neurogenic disease (neuropathy), or the like, and may be a myogenic disease in particular.
具体的な筋疾患としては、例えば、筋萎縮[例えば、筋原性の筋萎縮(例えば、筋ジストロフィーなど)、神経原性の筋萎縮(例えば、脊髄性筋萎縮症(SMA)、球脊髄性筋萎縮症など)など]、筋細胞の異常を伴う疾患(例えば、筋ジストロフィー、筋硬直性疾患)、エネルギー代謝の異常を伴う疾患(例えば、ミトコンドリア脳筋症、周期性四肢麻痺など)、神経筋接合部疾患(例えば、重症筋無力症、ランバートイートン(Lambert−Eaton)症候群など)などが挙げられる。 Specific muscle diseases include, for example, muscular atrophy [eg, myogenic muscular dystrophy (eg, muscular dystrophy), neurogenic muscular atrophy (eg, spinal muscular atrophy (SMA), bulbar muscular muscle). Atrophy, etc.)], diseases with abnormal muscle cells (eg, muscular dystrophy, muscular rigidity), diseases with abnormal energy metabolism (eg, mitochondrial encephalomyopathy, periodic limb palsy, etc.), neuromuscular junction Partial diseases (eg, severe muscular dystrophy, Lambert-Eaton syndrome, etc.) and the like can be mentioned.
また、筋疾患(筋萎縮)には、神経疾患に伴うもの(例えば、脊髄損傷による筋萎縮などの前記神経疾患に伴う筋疾患又は筋萎縮)も含まれる。 The muscle disease (muscle atrophy) also includes those associated with a neurological disease (for example, muscle disease or muscle atrophy associated with the neurological disease such as muscle atrophy due to spinal cord injury).
サルコペニア、ロコモーティブシンドローム及び麻痺の要因(原因)は、特に限定されず、上記神経疾患(脊髄損傷など)や筋疾患(筋原性疾患など)によるものであってもよく、これらによらないものであってもよい。 The factors (causes) of sarcopenia, locomotive syndrome and paralysis are not particularly limited, and may be due to the above-mentioned neurological diseases (spinal cord injury, etc.) and muscle diseases (myogenic diseases, etc.), and are not based on these. There may be.
さらに、第2の態様の剤又は組成物は、筋細胞の増殖や筋肉量の増加(さらには軸索の伸展)を促すため、何らかの症状・疾患に至っているか否かを問わず、運動機能や体力を改善ないし維持しうる。そのため、第2の態様の剤又は組成物は、(6)運動機能及び/又は体力の改善(向上)及び/又は維持のために使用してもよい。 Furthermore, the agent or composition of the second aspect promotes the proliferation of muscle cells and the increase of muscle mass (furthermore, the extension of axons), so that the motor function and the motor function may be affected regardless of whether or not any symptom / disease is reached. Can improve or maintain physical fitness. Therefore, the agent or composition of the second aspect may be used for (6) improvement (improvement) and / or maintenance of motor function and / or physical strength.
例えば、第2の態様の剤又は組成物は、(i)上記(1)〜(4)のような具体的疾患・症状にある対象(ヒト)の運動機能及び/又は体力を改善(向上)及び/又は維持するために使用してもよく、(ii)上記(1)〜(4)以外の疾患・症状にある対象(ヒト)の運動機能及び/又は体力を改善(向上)及び/又は維持するために使用してもよく、(iii)何らかの疾患・症状にない対象(ヒト)の運動機能及び/又は体力を改善(向上)及び/又は維持するため(例えば、老化等に伴う体力の減少の改善及び/向上など)に使用してもよい。 For example, the agent or composition of the second aspect improves (improves) the motor function and / or physical strength of a subject (human) having a specific disease / symptom as described in (i) above (1) to (4). And / or may be used to maintain (ii) improve (improve) and / or physical fitness of a subject (human) with a disease / symptom other than (1) to (4) above. It may be used to maintain (iii) to improve (improve) and / or maintain the motor function and / or physical strength of a subject (human) who does not have any disease / symptom (for example, physical strength associated with aging, etc.) It may be used to improve and / improve the decrease.
なお、上記の第1の態様で記載したように、成分(A)は、ペリオスチンやPKM2の分泌により(分泌を介して)、筋細胞の増殖、筋肉量の増加や、軸索の伸展を実現できる。 As described in the first aspect above, the component (A) realizes the proliferation of muscle cells, the increase in muscle mass, and the extension of axons by secreting periostin and PKM2 (via secretion). it can.
そのため、第2の態様において、筋細胞の増殖(増殖の促進)、筋肉量の増加及び/又は筋萎縮の改善[さらには、このような機能に関連した上記種々の用途((1)〜(6)など)]は、ペリオスチン及び/又はPKM2の分泌を通じて(介して)行われてもよい。 Therefore, in the second aspect, muscle cell proliferation (promotion of proliferation), increase in muscle mass and / or improvement of muscular atrophy [Furthermore, the above-mentioned various uses related to such functions ((1) to (1) to ( 6) etc.)] may be performed through (via) the secretion of periostin and / or PKM2.
本発明の剤又は組成物は、上記のように、種々の症状又は疾患を改善しうるが、このような症状又は疾患(例えば、神経疾患、筋疾患、サルコペニア、ロコモーティブシンドローム、麻痺など)は、慢性期のもの(慢性疾患、慢性症状)であってもよい。なお、慢性期とは、特に限定されず、症状又は疾患等によるが、通常、変動期(急性期、症状が激しい時期)を経過後の状態を言う場合が多い。例えば、脊髄損傷(ヒトの場合)であれば、重傷してから数ヶ月後には、慢性期になる場合が多い。 The agents or compositions of the present invention may ameliorate various symptoms or diseases as described above, but such symptoms or diseases (eg, neurological disorders, muscle disorders, sarcopenia, locomotive syndrome, paralysis, etc.) It may be in the chronic phase (chronic disease, chronic symptom). The chronic phase is not particularly limited and depends on the symptom or disease, but usually refers to the state after the lapse of the variable phase (acute phase, severe symptom). For example, in the case of spinal cord injury (in the case of humans), it is often the chronic phase several months after the serious injury.
通常、慢性期の症状の治療や改善は困難であるが、本発明の剤又は組成物によれば、慢性期の疾患又は症状(例えば、脊髄損傷、脊髄損傷に伴う筋疾患)であっても、効率よく治療及び/又は改善しうる。 It is usually difficult to treat or ameliorate chronic symptoms, but according to the agents or compositions of the invention, even chronic diseases or symptoms (eg, spinal cord injury, muscle disease associated with spinal cord injury) Can be treated and / or improved efficiently.
[成分(A)]
本発明の剤又は組成物(第1の態様及び第2の態様)は、アクテオシド、エキナコシド及びこれらの塩(特に薬学的に許容される塩)から選択された少なくとも1種の成分(A)を含む。[Component (A)]
The agent or composition of the present invention (first aspect and second aspect) contains at least one component (A) selected from acteosides, echinacosides and salts thereof (particularly pharmaceutically acceptable salts). Including.
塩としては、特に限定されず、例えば、金属塩[例えば、アルカリ金属塩(例えば、リチウム塩、ナトリウム塩、カリウム塩など)、アルカリ土類金属塩(例えば、マグネシウム塩、カルシウム塩)、周期表第13族金属塩(例えば、アルミニウム塩)、遷移金属塩(例えば、亜鉛などの塩)など]、アンモニウム塩、アミン塩[例えば、アルキルアミン塩(例えば、トリメチルアミン塩、トリエチルアミン塩のトリアルキルアミン塩)、アルカノールアミン塩(例えば、モノエタノールアミン塩、トリエタノールアミン塩)、環式アミン塩(例えば、ピリジン塩)]などが挙げられる。 The salt is not particularly limited, and for example, a metal salt [for example, an alkali metal salt (for example, lithium salt, sodium salt, potassium salt, etc.), an alkaline earth metal salt (for example, magnesium salt, calcium salt, etc.), a periodic table. Group 13 metal salts (eg aluminum salts), transition metal salts (eg salts such as zinc)], ammonium salts, amine salts [eg alkylamine salts (eg trimethylamine salts, trialkylamine salts of triethylamine salts) ), Alkanolamine salt (for example, monoethanolamine salt, triethanolamine salt), cyclic amine salt (for example, pyridine salt)] and the like.
特に、成分(A)は、少なくともアクテオシド[Acteoside、ベルバスコシド(Verbascoside)、クサギニン(Kusaginin)]を含んでいてもよい。 In particular, the component (A) may contain at least acteoside [Acteoside, Verbascoside, Kusaginin].
なお、成分(A)が、アクテオシド及びエキナコシドを含む場合、これらの割合は、特に限定されないが、例えば、アクテオシド/エキナコシド(質量比)=100/1〜0.01/1、好ましくは50/1〜0.02/1(例えば、30/1〜0.03/1)、さらに好ましくは20/1〜0.05/1(例えば、15/1〜0.07/1)程度であってもよく、10/1〜0.1/1(例えば、1/1〜0.1/1、0.9/1〜0.2/1、0.8/1〜0.25/1、0.7/1〜0.3/1など)であってもよい。 When the component (A) contains an acteoside and an echinacoside, the ratio thereof is not particularly limited, but for example, acteoside / echinakoside (mass ratio) = 100/1 to 0.01/1, preferably 50/1. Even if it is about 0.02 / 1 (for example, 30/1 to 0.03 / 1), more preferably 20/1 to 0.05 / 1 (for example, 15/1 to 0.07 / 1). Often, 10 / 1-0.1 / 1 (eg, 1/1-0.1 / 1, 0.9 / 1-0.2 / 1, 0.8 / 1-0.25 / 1, 0. 7 / 1-0.3 / 1 etc.).
なお、成分(A)は、合成したものを用いてもよく、天然物由来であってもよい。なお、合成方法は、慣用の手法を利用できる。 The component (A) may be a synthetic product or may be derived from a natural product. As the synthesis method, a conventional method can be used.
例えば、アクテオシドは、ハマウツボ科の植物(例えば、ニクジュウヨウ(肉従蓉)など)、ジオウ、オリーブ、ヘラオオバコなどに含まれており、このような植物から得てもよい。 For example, acteosides are found in plants of the family Orobanche coeruleus (eg, Nikjuyo (meat servant)), dioe, olives, ribwort plantain, etc., and may be obtained from such plants.
また、エキナコシド(echinacoside)は、ハマウツボ科の植物(例えば、ニクジュウヨウ(肉従蓉)など)、エキナセア アングスティフォリア(Echinacea angustifolia)、エキナセア パリダ(Echinacea pallida)、エキナセア プルプレア(Echinacea purpurea)などに含まれている。 Echinacoside is a plant of the Echinacea family (for example, Echinacea angustifolia), Echinacea angustifolia, Echinacea pallida, Echinacea purpurea, etc. ing.
ハマウツボ科の植物、特に、ニクジュウヨウには、アクテオシド(さらにはエキナコシド)に加えて、その他の成分を含んでいる場合が多く、これらを組み合わせて有することと相まってか、本発明の効果を有効に発現しやすい。また、安全性の点においても、ニクジュウヨウ等を好適に使用しうる。 Plants of the family Broomrape, especially Nikujuyou, often contain other components in addition to acteosides (and echinacosides), and in combination with having these components, the effects of the present invention are effectively effective. Easy to develop. Further, in terms of safety, Nikujuyo and the like can be preferably used.
そのため、本発明の剤又は組成物は、成分(A)を含有する成分として、特に、ニクジュウヨウ成分などのハマウツボ科植物の成分を含んでいてもよい。 Therefore, the agent or composition of the present invention may contain, as a component containing the component (A), a component of a broomrape family plant such as a component of Orobanche coeruleus.
ハマウツボ科の植物の成分としては、ハマウツボ科植物、その抽出物(エキス)などが挙げられ、これらを組み合わせてもよい。具体的には、ニクジュウヨウ成分としては、ニクジュウヨウ、ニクジュウヨウの抽出物(ニクジュウヨウエキス)などが挙げられ、これらを組み合わせてもよい。 Examples of the components of the broomrape family plant include broomrape family plants and their extracts (extracts), and these may be combined. Specifically, examples of the Nikujuyo component include Nikujuyo, an extract of Nikujuyo (Nikujuyo extract), and the like, and these may be combined.
ハマウツボ科の植物は、根、茎、幹、樹皮、葉、花、果実、果皮、果穂、種子、種皮などの部分を使用してもよく、これらを組み合わせて使用してもよく、全草を使用してもよい。特に、少なくとも茎(肉質茎)を好適に使用してもよい。 Plants of the family Broomrape may use parts such as roots, stems, trunks, bark, leaves, flowers, fruits, pericarps, spikes, seeds, seed coats, etc., or may be used in combination thereof, and the whole plant may be used. You may use it. In particular, at least stems (fleshy stems) may be preferably used.
なお、ニクジュウヨウとしては、特に限定されないが、ホンオニク(Cistanche salsa)、ジュウヨウ(Cistanche deserticola)及び/又はカンカニクジュウヨウ(Cistanche tubulosa)の茎(肉質茎)が挙げられる。
これらの中でも、カンカニクジュウヨウを好適に使用してもよい。The cistanche salsa is not particularly limited, and examples thereof include cistanche salsa, cistanche deserticola and / or cistanche tubulosa stalks (flesh stalks).
Among these, Kankanikujuyo may be preferably used.
エキスは、市販品を利用してもよく、ハマウツボ科の植物(ニクジュウヨウなど)を慣用の方法で抽出処理{例えば、抽出溶媒[例えば、水、アルコール(メタノール、エタノールなど)、これらの混合物など]を用い、常温または加熱下で常法またはそれに準じた方法で抽出処理}することで得ることもできる。 As the extract, a commercially available product may be used, and a plant of the family Broomrape (Nikujuyou, etc.) is extracted by a conventional method {for example, an extraction solvent [for example, water, alcohol (methanol, ethanol, etc.), a mixture thereof, etc. ], It can also be obtained by extraction treatment at room temperature or heating by a conventional method or a method similar thereto}.
抽出物の中でも、特に、成分(A)の含量や濃度が高い部分を分離又は分画して用いてもよい。 Among the extracts, a portion having a particularly high content or concentration of the component (A) may be separated or fractionated and used.
抽出に際しては、ハマウツボ科の植物(ニクジュウヨウなど)をそのまま又は粉砕(破砕)して供してもよい。また、抽出物は、さらに、濃縮や乾燥(常温乾燥、凍結乾燥など)に供してもよい。 At the time of extraction, a plant of the family Broomrape (such as Broomrape) may be used as it is or crushed (crushed). In addition, the extract may be further subjected to concentration and drying (normal temperature drying, freeze drying, etc.).
ハマウツボ科の植物の成分(ニクジュウヨウ成分など)の形態は、特に限定されないが、例えば、粉状(又は粉粒状)であってもよい。 The form of the component of the plant of the family Broomrape (such as the component of Orobanche coeruleus) is not particularly limited, but may be powdery (or powdery or granular), for example.
[形態・投与]
本発明の剤又は組成物の形態としては、特に限定されず、成分(A)[及び/又は後述の成分(B)]をそのまま剤(例えば、粉剤)又は組成物としてもよく、他の成分(担体、賦形剤など)とともに製剤化してもよい。[Form / administration]
The form of the agent or composition of the present invention is not particularly limited, and the component (A) [and / or the component (B) described later] may be used as it is as an agent (for example, a powder) or a composition, and other components. It may be formulated together with (carrier, excipient, etc.).
他の成分としては、一般に製剤の原料として用いられる成分等が挙げられ、特に限定されず、例えば、担体、賦形剤、結合剤、崩壊剤、滑沢剤、コーティング剤、着色剤、矯味矯臭剤、安定化剤、乳化剤、吸収促進剤、界面活性剤、pH調製剤、防腐剤、抗酸化剤などが挙げられる。これらの他の成分は、単独で又は2種以上組み合わせて使用してもよい。 Examples of other components include components generally used as raw materials for preparations, and are not particularly limited. For example, carriers, excipients, binders, disintegrants, lubricants, coating agents, colorants, and flavoring agents. Examples thereof include agents, stabilizers, emulsifiers, absorption promoters, surfactants, pH adjusters, preservatives, antioxidants and the like. These other components may be used alone or in combination of two or more.
担体としては、特に限定されないが、例えば、大豆油、牛脂、合成グリセライドなどの動植物油;流動パラフィン、スクワラン、固形パラフィンなどの炭化水素;ミリスチン酸オクチルドデシル、ミリスチン酸イソプロピルなどのエステル油;セトステアリルアルコール、ベヘニルアルコールなどの高級アルコール;シリコン樹脂;シリコーンオイル;ポリオキシエチレン脂肪酸エステル、ソルビタン脂肪酸エステル、グリセリン脂肪酸エステル、ポリオキシエチレンソルビタン脂肪酸エステル、ポリオキシエチレン硬化ひまし油、ポリオキシエチレンポリオキシプロピレンブロックコポリマーなどの界面活性剤;ヒドロキシエチルセルロース、ポリアクリル酸、カルボキシビニルポリマー、ポリエチレングリコール、ポリビニルピロリドン、メチルセルロースなどの水溶性高分子;エタノール、イソプロパノールなどの低級アルコール;グリセリン、プロピレングリコール、ジプロピレングリコール、ソルビトールなどの多価アルコール;グルコース、ショ糖などの糖;無水ケイ酸、ケイ酸アルミニウムマグネシウム、ケイ酸アルミニウムなどの無機粉体;精製水などが挙げられる。 The carrier is not particularly limited, and is, for example, animal and vegetable oils such as soybean oil, beef fat, and synthetic glyceride; hydrocarbons such as liquid paraffin, squalane, and solid paraffin; ester oils such as octyldodecyl myristate and isopropyl myristate; and cetostearyl. Higher alcohols such as alcohol and behenyl alcohol; Silicon resin; Silicone oil; Polyoxyethylene fatty acid ester, sorbitan fatty acid ester, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene block copolymer, etc. Surfactants; water-soluble polymers such as hydroxyethyl cellulose, polyacrylic acid, carboxyvinyl polymers, polyethylene glycol, polyvinylpyrrolidone, methyl cellulose; lower alcohols such as ethanol, isopropanol; glycerin, propylene glycol, dipropylene glycol, sorbitol, etc. Polyhydric alcohols; sugars such as glucose and sucrose; inorganic powders such as silicic anhydride, aluminum magnesium silicate, and aluminum silicate; purified water and the like.
賦形剤としては、例えば、乳糖、コーンスターチ、白糖、ブドウ糖、マンニトール、ソルビット、結晶セルロース、二酸化ケイ素などが挙げられる。 Examples of excipients include lactose, cornstarch, sucrose, glucose, mannitol, sorbitol, crystalline cellulose, silicon dioxide and the like.
結合剤としては、例えば、ポリビニルアルコール、ゼラチン、メチルセルロース、エチルセルロース、アラビアゴム、トラガント、ゼラチン、シェラック、ヒドロキシプロピルメチルセルロース、ヒドロキシプロピルセルロース、ポリビニルピロリドン、ポリビニルアセタールジエチルアミノアセテート、トウモロコシデンプンなどが挙げられる。 Examples of the binder include polyvinyl alcohol, gelatin, methyl cellulose, ethyl cellulose, gum arabic, tragant, gelatin, shelac, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, polyvinyl acetal diethylaminoacetate, corn starch and the like.
崩壊剤としては、例えば、トウモロコシデンプン、低置換度ヒドロキシプロピルセルロース、クロスポビドン、結晶セルロース、沈降炭酸カルシウム、クロスカルメロースナトリウム、クエン酸カルシウム、デキストリン、ペクチン、カルボキシメチルセルロースカルシウムなどが挙げられる。 Examples of the disintegrant include corn starch, low-substituted hydroxypropyl cellulose, crospovidone, crystalline cellulose, precipitated calcium carbonate, croscarmellose sodium, calcium citrate, dextrin, pectin, calcium carboxymethyl cellulose and the like.
滑沢剤としては、例えば、ステアリン酸マグネシウム、タルク、ポリエチレングリコール、軽質無水ケイ酸、ショ糖脂肪酸エステルなどが挙げられ、矯味矯臭剤としては、ココア末、メントール、芳香散、ハッカ油、竜脳、桂皮末などが用いられる。 Examples of the lubricant include magnesium stearate, talc, polyethylene glycol, light anhydrous silicic acid, sucrose fatty acid ester, and the like, and examples of the flavoring agent include cocoa powder, menthol, aromatic powder, peppermint oil, and dragon brain. Cinnamon powder is used.
剤又は組成物の形態(剤形)としては、特に限定されず、例えば、錠剤、散剤、細粒剤、顆粒剤、ドライシロップ剤、被覆錠剤、口腔内崩壊錠、チュアブル錠、カプセル剤、ソフトカプセル剤、シロップ剤、経口液剤、トローチ剤、ゼリー剤、吸入剤、坐剤、注射剤、軟膏剤、点眼剤、眼軟膏剤、点鼻剤、点耳剤、パップ剤、ローション剤、外用液剤、スプレー剤、外用エアゾール剤、クリーム剤、ゲル剤、テープ剤、バッカル錠、舌下錠、膣坐剤、膣錠、直腸ソフトカプセル剤等が挙げられる。 The form (dosage form) of the agent or composition is not particularly limited, and for example, tablets, powders, fine granules, granules, dry syrups, coated tablets, orally disintegrating tablets, chewable tablets, capsules, soft capsules. , Syrups, oral solutions, troches, jellies, inhalants, suppositories, injections, ointments, eye drops, eye ointments, nasal drops, ear drops, poultices, lotions, topical solutions, sprays Examples thereof include agents, topical aerosols, creams, gels, tapes, buccal tablets, sublingual tablets, vaginal suppositories, vaginal tablets, and rectal soft capsules.
剤又は組成物の投与(服用)形態は、特に限定されず、経口投与であっても、非経口投与であってもよい。非経口投与としては、例えば、直腸投与、経鼻投与、経肺投与、注射投与(例えば、静脈内投与、脊椎腔内投与、硬膜外腔内投与、筋肉内投与、皮下投与、腹腔内投与、動脈内投与、関節内投与、心臓内投与、嚢内投与、皮内投与、病巣内投与、眼内投与、胸腔内投与、くも膜下投与、子宮内投与、脳室内投与)などが挙げられる。 The form of administration (administration) of the agent or composition is not particularly limited, and may be oral administration or parenteral administration. Parenteral administration includes, for example, rectal administration, nasal administration, pulmonary administration, injection administration (for example, intravenous administration, intravertebral administration, intraepidural administration, intramuscular administration, subcutaneous administration, intraperitoneal administration). , Intra-arterial administration, intra-arterial administration, intracardiac administration, intracapsular administration, intradermal administration, intralesional administration, intraocular administration, intrathoracic administration, epidural administration, intrauterine administration, intraventricular administration) and the like.
特に、成分(A)[及び/又は後述の成分(B)]は、前記のように、筋細胞の増殖や筋肉量の増加を伴う場合が多いため、本発明の剤又は組成物は、筋肉内投与を好適に適用してもよい。一方で、簡便な投与という観点から、経口投与を好適に使用してもよい。 In particular, since the component (A) [and / or the component (B) described later] often accompanies the proliferation of muscle cells and the increase in muscle mass as described above, the agent or composition of the present invention is a muscle. Oral administration may be preferably applied. On the other hand, oral administration may be preferably used from the viewpoint of simple administration.
本発明の剤又は組成物において、成分(A)[及び/又は後述の成分(B)]の量は、特に限定されず、剤形、投与量等に応じて適宜選択できる。 In the agent or composition of the present invention, the amount of the component (A) [and / or the component (B) described later] is not particularly limited and can be appropriately selected depending on the dosage form, dosage and the like.
本発明の剤又は組成物の投与量(摂取量、服用量)は、疾患・症状の程度、年齢、性別、体重、投与形態、塩の種類、疾患の具体的な種類等に応じて異なるが、例えば、1日あたり、成分(A)の量として、0.4〜40mg/kg、好ましくは2〜20mg/kg、さらに好ましくは4〜8mg/kg程度であってもよい。 The dose (intake, dose) of the agent or composition of the present invention varies depending on the degree of disease / symptom, age, sex, body weight, administration form, type of salt, specific type of disease, and the like. For example, the amount of the component (A) per day may be about 0.4 to 40 mg / kg, preferably about 2 to 20 mg / kg, and more preferably about 4 to 8 mg / kg.
特に、成分(A)を含む成分としてニクジュウヨウエキスを使用する場合、ニクジュウヨウエキスの投与量は、例えば、1〜6000mg/kg、好ましくは5〜3000mg/kg、さらに好ましくは10〜1200mg/kg程度であってもよい。 In particular, when Nikujuyo extract is used as a component containing the component (A), the dose of Nikujuyo extract is, for example, 1 to 6000 mg / kg, preferably 5 to 3000 mg / kg, and more preferably 10 to 1200 mg / kg. It may be about.
投与は、1回又は複数回に分けてもよい。 Administration may be divided into single doses or multiple doses.
本発明の剤又は組成物は、医薬又は医薬組成物であってもよく、医薬部外品であってもよい。 The agent or composition of the present invention may be a drug or a pharmaceutical composition, or may be a quasi drug.
また、本発明の剤又は組成物は、飲食品の形態であってもよい。そのため、本発明には、成分(A)[及び/又は後述の成分(B)](又は前記剤又は組成物)を含む飲食品も含まれる。また、成分(A)[及び/又は後述の成分(B)](又は前記剤又は組成物)という観点からは、成分(A)[及び/又は後述の成分(B)]は、飲食品の添加剤であってもよい。 Moreover, the agent or composition of this invention may be in the form of food and drink. Therefore, the present invention also includes foods and drinks containing the component (A) [and / or the component (B) described later] (or the agent or composition). Further, from the viewpoint of the component (A) [and / or the component (B) described later] (or the agent or composition), the component (A) [and / or the component (B) described later] is a food or drink. It may be an additive.
なお、このような飲食品において、成分(A)の好ましい態様などは、前記と同様であってもよい。 In such foods and drinks, the preferred embodiment of the component (A) may be the same as described above.
飲食品としては、いわゆる健康食品を含む一般食品の他、厚生労働省の保健機能食品制度に規定された特定保健用食品や栄養機能食品などの保健機能食品をも含むものであり、さらに、サプリメント(栄養補助食品)、飼料、食品添加物なども本発明の飲食品に包含される。 Foods and drinks include general foods including so-called health foods, as well as health functional foods such as foods for specified health use and nutritional functional foods stipulated in the health functional food system of the Ministry of Health, Labor and Welfare, and supplements ( Dietary supplements), feeds, food additives and the like are also included in the foods and drinks of the present invention.
また、飲食品は、特定の対象者用[例えば、高齢者用、患者又は病者用(例えば、神経疾患、筋疾患、サルコペニア、ロコモーティブシンドローム、麻痺などの(前記例示の疾患又は症状など)を有する患者用)など]の飲食品であってもよい。 In addition, foods and drinks are intended for specific subjects [for example, for elderly people, patients or sick people (for example, neurological diseases, muscle diseases, sarcopenia, locomotive syndrome, paralysis, etc. (such as the above-exemplified diseases or symptoms)). (For patients who have), etc.] may be food and drink.
成分(A)を飲食品に使用するには、そのまま、または種々の栄養成分等とともに加工肉、清涼飲料などの飲食品の原料に混ぜて飲食品を製造することができる。 In order to use the component (A) in a food or drink, the food or drink can be produced as it is or mixed with various nutritional components and the like as raw materials for the food and drink such as processed meat and soft drinks.
また、成分(A)を、健康食品、栄養補助食品などとして使用する場合、例えば、慣用の手段を用いて、錠剤、カプセル(ソフトカプセル、ハードカプセルなど)、散剤、顆粒、液剤(懸濁剤、シロップ剤など)、乳剤、ゼリー、スティック状などの形態に調製することができる。また、錠剤には、崩壊錠(口腔内崩壊錠)も含まれる。 When the ingredient (A) is used as a health food, dietary supplement, etc., for example, tablets, capsules (soft capsules, hard capsules, etc.), powders, granules, liquids (suspensions, syrups) are used by conventional means. Agents, etc.), emulsions, jellies, sticks, etc. can be prepared. The tablets also include disintegrating tablets (orally disintegrating tablets).
飲食品は、食品添加剤(食品用添加剤)を含んでいてもよい。食品添加剤としては、特に限定されないが、例えば、賦形剤(例えば、コムギデンプン、トウモロコシデンプン、セルロース、乳糖、ショ糖、マンニトール、ソルビトール、キシリトール、アルファー化デンプン、カゼイン、ケイ酸アルミン酸マグネシウム、ケイ酸カルシウムなど)、結合剤(例えば、アルファー化デンプン、ヒドロキシプロピルメチルセルロース、ポリビニルピロリドンなど)、崩壊剤(例えば、セルロース、ヒドロキシプロピルセルロース、トウモロコシデンプンなど)、流動化剤(例えば、軽質無水ケイ酸、ショ糖脂肪酸エステルなど)、油(例えば、大豆油、ゴマ油、オリーブ油、亜麻仁油、エゴマ油、ナタネ油、ココナッツ油、トウモロコシ油などの植物油又は動物・魚由来の油)、栄養素(例えば、各種ミネラル、各種ビタミン、アミノ酸)、香料、甘味料、矯味剤、着色料、溶媒(エタノール)、塩類、界面活性剤、pH調節剤、緩衝剤、抗酸化剤、安定化剤、ゲル化剤、増粘剤、滑沢剤、カプセル化剤、懸濁剤、コーティング剤、防腐剤などが挙げられる。 Foods and drinks may contain food additives (food additives). The food additive is not particularly limited, but for example, excipients (for example, wheat starch, corn starch, cellulose, lactose, sucrose, mannitol, sorbitol, xylitol, pregelatinized starch, casein, magnesium silicate aluminate, etc. Calcium silicate, etc.), excipients (eg, pregelatinized starch, hydroxypropylmethylcellulose, polyvinylpyrrolidone, etc.), disintegrants (eg, cellulose, hydroxypropylcellulose, corn starch, etc.), fluidizers (eg, light anhydrous silicic acid) , Sucrose fatty acid esters, etc.), oils (eg, soybean oil, sesame oil, olive oil, flaxseed oil, sesame oil, rapeseed oil, coconut oil, corn oil and other vegetable oils or animal / fish-derived oils), nutrients (eg, various Minerals, various vitamins, amino acids), fragrances, sweeteners, flavoring agents, coloring agents, solvents (ethanol), salts, surfactants, pH adjusters, buffers, antioxidants, stabilizers, gelling agents, increase Examples include adhesives, starches, encapsulants, suspending agents, coating agents, preservatives and the like.
食品添加剤は、単独で又は2種以上組み合わせて使用してもよい。 The food additive may be used alone or in combination of two or more.
成分(A)を飲食品添加剤として用いる場合、飲食品としては、特に限定されないが、例えば、食品[例えば、麺類(そば、うどん、中華麺、即席麺など)、菓子類(飴、キャンディー、ガム、チョコレート、スナック菓子(ポテトチップなど)、ビスケット、クッキー、グミ、ゼリー、ジャム、バター、クリーム(シュークリームなど)、ケーキなど)、パン類、水産又は畜産加工食品(かまぼこ、ハム、ソーセージなど)、乳製品(加工乳、発酵乳など)、油脂および油脂加工食品(サラダ油、てんぷら油、マーガリン、マヨネーズ、ショートニング、ホイップクリーム、ドレッシングなど)、調味料(ソース、たれなど)、レトルト食品(カレー、シチュー、丼、お粥、雑炊など)、冷菓(アイスクリーム、シャーベット、かき氷など)、揚げ物(コロッケ、フライドポテト、フライドチキンなど)など]、飲料(茶飲料、清涼飲料、炭酸飲料、栄養飲料、果実飲料、乳酸飲料など)などが挙げられる。 When the component (A) is used as a food and drink additive, the food and drink are not particularly limited, but for example, foods [for example, noodles (soba, udon, Chinese noodles, instant noodles, etc.), confectionery (candy, candy, etc.) Gum, chocolate, snacks (potato chips, etc.), biscuits, cookies, gummy, jelly, jam, butter, cream (cream puffs, etc.), cakes, breads, marine or processed livestock foods (kamaboko, ham, sausage, etc.), Dairy products (processed milk, fermented milk, etc.), fats and oils and processed fats and oils (salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressings, etc.), seasonings (sauce, sauce, etc.), retort foods (curry, stew) , Bowls, porridge, miscellaneous dishes, etc.), cold confectionery (ice cream, sorbet, shaved ice, etc.), fried foods (croquette, fried potatoes, fried chicken, etc.)], beverages (tea beverages, soft drinks, carbonated beverages, nutritional beverages, fruits Beverages, lactic acid beverages, etc.).
上記の飲食品における成分(A)の配合量は、添加形態及び投与形態によっても異なり、広い範囲から選択できるが、例えば、0.0001重量%以上(例えば、0.001〜50重量%)、0.003重量%以上(例えば、0.005〜30重量%)、0.01重量%以上(例えば、0.05〜10重量%)などであってもよい。 The blending amount of the component (A) in the above food and drink varies depending on the addition form and the administration form and can be selected from a wide range. For example, 0.0001% by weight or more (for example, 0.001 to 50% by weight). It may be 0.003% by weight or more (for example, 0.005 to 30% by weight), 0.01% by weight or more (for example, 0.05 to 10% by weight), and the like.
なお、飲食品において、成分(A)の摂取量(又は投与量又は服用量)は、前記と同様の範囲から選択できる。 In the food and drink, the intake (or dose or dose) of the component (A) can be selected from the same range as described above.
<方法、その他の剤又は組成物>
本発明の剤又は組成物(又は成分(A))は、上記の通り、種々の用途に適用可能である。そのため、本発明には、当該用途に対応した方法も包含する。<Method, other agent or composition>
As described above, the agent or composition (or component (A)) of the present invention can be applied to various uses. Therefore, the present invention also includes a method corresponding to the said use.
まず、第1の方法では、成分(A)(又は前記剤又は組成物)により、ペリオスチン及び/又はPKM2を分泌させる(ペリオスチン及び/又はPKM2の分泌を促進させる)。 First, in the first method, the component (A) (or the agent or composition) causes the secretion of periostin and / or PKM2 (promotes the secretion of periostin and / or PKM2).
なお、前記のように、ペリオスチン及び/又はPKM2により、筋細胞が増殖したり、筋肉が増加したり、軸索が伸展又は修復する。そのため、本発明には、下記発明も包含する。 As described above, periostin and / or PKM2 proliferates muscle cells, increases muscles, and stretches or repairs axons. Therefore, the present invention also includes the following inventions.
(i)ペリオスチン及びPKM2から選択された少なくとも1種の成分(B)を含み、筋細胞の増殖、筋肉の増加(増大)、及び/又は筋萎縮の改善(治療)のための剤又は組成物。 (I) An agent or composition containing at least one component (B) selected from periostin and PKM2 for improving (treating) muscle cell proliferation, muscle gain (growth), and / or muscular atrophy. ..
(ii)ペリオスチン及びPKM2から選択された少なくとも1種の成分(B)を含み、軸索を伸展及び/又は修復するための剤又は組成物。 (Ii) An agent or composition containing at least one component (B) selected from periostin and PKM2 for extending and / or repairing axons.
(iii)ペリオスチン及びPKM2から選択された少なくとも1種の成分(B)を用いて(又は成分(B)を分泌させ)、筋細胞を増殖させる、筋肉を増加(増大)させる及び/又は筋萎縮を改善(治療)する方法。 (Iii) Using at least one component (B) selected from periostin and PKM2 (or secreting component (B)), muscle cells proliferate, muscle gain (growth) and / or muscle atrophy How to improve (treat).
(iv)ペリオスチン及びPKM2から選択された少なくとも1種の成分(B)を用いて(又は成分(B)を分泌させ)、軸索を伸展及び/又は修復する方法。 (Iv) A method of extending and / or repairing an axon using (or secreting) a component (B) of at least one selected from periostin and PKM2.
これらの剤又は組成物や方法では、前記のように、これらに対応する疾患や症状の改善(例えば、下記(1)〜(6))などに利用できる。 As described above, these agents, compositions and methods can be used for improving diseases and symptoms corresponding to them (for example, (1) to (6) below).
(1)神経疾患の治療及び/又は改善
(2)筋疾患の治療及び/又は改善
(3)サルコペニアの治療及び/又は改善
(4)ロコモーティブシンドロームの治療及び/又は改善
(5)麻痺の治療及び/又は改善
(6)運動機能及び/又は体力の改善(向上)及び/又は維持(1) Treatment and / or improvement of neurological disorders (2) Treatment and / or improvement of muscle diseases (3) Treatment and / or improvement of sarcopenia (4) Treatment and / or improvement of locomotive syndrome (5) Treatment and improvement of paralysis / Or improvement (6) Improvement (improvement) and / or maintenance of motor function and / or physical strength
このような剤又は組成物及び方法において、ペリオスチンやPKM2は、生体内に分泌させたものであってもよく、生体外に存在するもの(生体外にて合成したものなど)であってもよい。例えば、ペリオスチンやPKM2は、リコンビナント(組換え体)であってもよく、有機化学的手法により合成されたものであってもよい。
また、上記剤又は組成物及び方法では、成分(B)を少なくとも使用すればよく、成分(A)と成分(A)とを併用してもよい。In such agents, compositions and methods, periostin and PKM2 may be secreted in vivo or may be in vitro (such as those synthesized in vitro). .. For example, periostin and PKM2 may be recombinants or may be synthesized by an organic chemical method.
Further, in the above-mentioned agent, composition and method, at least the component (B) may be used, and the component (A) and the component (A) may be used in combination.
第2の方法では、成分(A)(又は前記剤又は組成物)を用いて(例えば、投与し)、筋細胞を増殖させる、筋肉を増加(増大)させる及び/又は筋萎縮を改善(治療)する。この方法では、さらに、軸索を伸展及び/又は修復させてもよい。そして、この方法は、例えば、これらに対応する疾患や症状の改善(例えば、前記(1)〜(6))などに利用できる。 In the second method, component (A) (or the agent or composition) is used (eg, administered) to proliferate muscle cells, increase (grow) muscle and / or improve (treat) muscle atrophy. ). In this method, the axons may also be extended and / or repaired. Then, this method can be used, for example, for improving diseases and symptoms corresponding to these (for example, (1) to (6) above).
なお、第2の方法では、ペリオスチン及び/又はPKM2の分泌を通じて(介して)、筋細胞を増殖させて、筋肉を増加(増大)させて及び/又は筋萎縮を改善(治療)してもよい。 In the second method, muscle cells may be proliferated through (via) secretion of periostin and / or PKM2 to increase (increase) muscle and / or improve (treat) muscle atrophy. ..
なお、上記剤又は組成物、方法等において、投与方法等の態様は、前記と同様であってもよい。
例えば、成分(B)の量(投与量、服用量)は、0.1〜50mg/kg、0.5〜10mg/kg、1〜5mg/kgなどであってもよい。より具体的な成分(B)の投与量として、0.1〜0.5mg/kg、1〜5mg/kg、10〜50mg/kgなどを選択することもできる。In the above-mentioned agent, composition, method, etc., the mode of administration method, etc. may be the same as described above.
For example, the amount (dose, dose) of the component (B) may be 0.1 to 50 mg / kg, 0.5 to 10 mg / kg, 1 to 5 mg / kg, or the like. As a more specific dose of the component (B), 0.1 to 0.5 mg / kg, 1 to 5 mg / kg, 10 to 50 mg / kg and the like can be selected.
以下に、実施例を挙げて本発明をさらに具体的に説明するが、本発明はこれらの実施例により何ら限定されるものではなく、多くの変形が本発明の技術的思想内で当分野において通常の知識を有する者により可能である。 Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples, and many modifications are made in the art within the technical idea of the present invention. It is possible by a person with ordinary knowledge.
<ニクジュウヨウエキス>
ニクジュウヨウエキスはニクジュウヨウ(栃本天海堂、大阪)に20倍量の水を加え、煎じ器(ウチダ和漢薬、東京)を用いて抽出を行った。生薬に加えた水が沸騰するまで強火で加熱し(10〜15分)、その後、とろ火での加熱を総加熱時間1時間になるまで続けた。ろ液を濾した後、熱時綿栓濾過し、ろ液を室温で放冷後、液体窒素で凍結させた。凍結乾燥器にて乾燥させた粉末をエキス粉末とし(収率40.8%)、生理食塩水に溶解してニクジュウヨウエキスを得た。
なお、ニクジュウヨウエキスにおいて、アクテオシドの含有量は0.21重量%、エキナコシドの含有量は0.35重量%であった。<Nikujuyo extract>
The Nikujuyo extract was extracted by adding 20 times the amount of water to Nikujuyo (Tochimoto Tenkaido, Osaka) and using a decoction machine (Uchida Wakanyaku, Tokyo). The water added to the crude drug was heated on high heat (10 to 15 minutes) until it boiled, and then heating on a low heat was continued until the total heating time reached 1 hour. After filtering the filtrate, it was filtered with a cotton swab when heated, and the filtrate was allowed to cool at room temperature and then frozen in liquid nitrogen. The powder dried in a freeze-dryer was used as an extract powder (yield 40.8%) and dissolved in physiological saline to obtain an extract of Nikujuyo.
In the Nikujuyo extract, the content of acteoside was 0.21% by weight, and the content of echinacoside was 0.35% by weight.
<アクテオシド>
アクテオシド(Acteoside、別名:verbascoside、kusaginin)は、市販品(Extrasynthesis社、仏国)を生理食塩水に溶解して使用した。<Acteoside>
Acteoside (also known as verbascoside, kusaginin) was used by dissolving a commercially available product (Extrasynthesis, France) in physiological saline.
<供試動物>
(1)脊髄損傷モデルマウス:ddYマウスを、日本SLC(浜松、日本)から得た。本実施例においては、雌性かつ8週齢のddYマウスを用いた。
(2)初代培養骨格筋細胞用のマウス:生後3日目のddYマウスを用いた。
(3)初代培養神経細胞用のマウス:胎生14日齢のddYマウスを用いた。
全てのマウスは、固形飼料と水を自由に摂取させ、22±2℃、50±5%の湿度、午前7時から始まる12時間の明暗サイクルで制御された環境で飼育した。<Test animal>
(1) Spinal cord injury model mouse: A ddY mouse was obtained from Japan SLC (Hamamatsu, Japan). In this example, female and 8-week-old ddY mice were used.
(2) Mice for primary cultured skeletal muscle cells: ddY mice 3 days after birth were used.
(3) Mice for primary cultured neurons: 14-day-old embryonic ddY mice were used.
All mice were fed free feed and water and bred in a controlled environment at 22 ± 2 ° C., 50 ± 5% humidity, and a 12-hour light-dark cycle starting at 7 am.
<マウス骨格筋細胞の初代培養>
生後3日目のddYマウスを麻酔下におき左右後肢の骨格筋全体を摘出した。細かく切断した筋組織を分解液(HBSS緩衝液中に、1mg/mL collagenase、0.02mg/mL dispase I、2.5mM塩酸カルシウム)に加え、37℃で1時間インキュベートした。遠心分離後の沈渣に、筋細胞増殖培地(Myocyte Growth Medium、Promo cell、独国)を加えて、パスツールピペットによるピペッティングにより細胞塊がほぼ見えなくなるまで細胞を穏やかに懸濁した後、70μmのナイロン製セルストレーナー(nylon cell strainer、Falcon、米国)で濾過した。細胞の培養は8ウェルのチャンバースライド(Falcon)、白色96ウェルプレート、6cm培養皿(Falcon)で行った。
細胞増殖活性を評価する場合には、白色96ウェルプレートに筋細胞を1,000cells/wellの密度で播種し、3日間の薬物処置後、CellTiter−Glo試薬(Promega、米国)によって生存細胞を測定した。<Primary culture of mouse skeletal muscle cells>
Three-day-old ddY mice were placed under anesthesia and the entire skeletal muscles of the left and right hind limbs were removed. Finely chopped muscle tissue was added to a degradation solution (1 mg / mL collagenase, 0.02 mg / mL dispase I, 2.5 mM calcium chloride in HBSS buffer) and incubated at 37 ° C. for 1 hour. Myocyte Growth Medium (Promo cell, Germany) was added to the sediment after centrifugation, and the cells were gently suspended by pipetting with a Pasteur pipette until the cell mass became almost invisible, and then 70 μm. Filtered with a nylon cell strainer (Falcon, USA). Cells were cultured on 8-well chamber slides (Falcon), white 96-well plates, and 6 cm culture dishes (Falcon).
To evaluate cell proliferation activity, muscle cells were seeded on a white 96-well plate at a density of 1,000 cells / well, and after 3 days of drug treatment, viable cells were measured with CellTiter-Glo reagent (Promega, USA). did.
<マウス大脳皮質神経細胞の初代培養>
胎生14日齢のddYマウス胎児の大脳皮質を安全キャビネット内で約1mm角に切断した。700rpmで3分間遠心した後、上清を除去し、沈殿物に0.05%トリプシン−0.53mM EDTA溶液(ライフテクノロジーズ)を2mL加え、懸濁した。37℃で15分間、5分置きに攪拌しながらインキュベーションし、培地を4mL加え、700rpmで3分間遠心して上清を除去し、沈殿に600U/mL DNase I(ライフテクノロジーズ)−0.03%トリプシンインヒビター(ライフテクノロジーズ)−PBS溶液を2mL加え、懸濁した。さらに、37℃で15分間、5分置きに攪拌しながらインキュベーションし、培地を4mL加え、700rpmで3分間遠心した。沈渣にカルシウム・マグネシウム不含HBSS(Hank’s balanced salt solution)を4mL加え700rpmで3分間遠心した。沈渣に培地を4mL加えた。先端を炙りなめしたパスツールピペットで細胞塊が見えなくなるまで穏やかに懸濁した後、70μmのナイロン製セルストレーナー(nylon cell strainer、Falcon)で濾過した。細胞の培養は8ウェルチャンバースライド(Falcon)で行った。前日にコーティング[Poly−D−リシン(Lysine)(PDL,Sigma-Aldrich)をPBSで0.005mg/mLに希釈したものをまき、37℃でインキュベーションしたもの。培養当日に滅菌精製水で2回洗浄。]したものに細胞を播種した。37℃、10%CO2、飽和水蒸気下で培養を開始し、5時間後に培地を全量、馬血清の代わりにB−27サプリメント(Invitrogen)を含む新しい培地で交換した。ならし培地(Conditioned medium)あるいは薬物処置後に、軸索伸展の評価においては免疫染色を実施した。<Primary culture of mouse cerebral cortex neurons>
The cerebral cortex of a 14-day-old ddY mouse fetus was cut into approximately 1 mm squares in a safety cabinet. After centrifuging at 700 rpm for 3 minutes, the supernatant was removed, and 2 mL of 0.05% trypsin-0.53 mM EDTA solution (Life Technologies) was added to the precipitate and suspended. Incubate at 37 ° C. for 15 minutes with stirring every 5 minutes, add 4 mL of medium, centrifuge at 700 rpm for 3 minutes to remove the supernatant, and deposit 600 U / mL DNase I (Life Technologies) -0.03% trypsin. 2 mL of Inhibitor (Life Technologies) -PBS solution was added and suspended. Further, the mixture was incubated at 37 ° C. for 15 minutes with stirring every 5 minutes, 4 mL of the medium was added, and the medium was centrifuged at 700 rpm for 3 minutes. 4 mL of calcium / magnesium-free HBSS (Hank's balanced salt solution) was added to the sediment and centrifuged at 700 rpm for 3 minutes. 4 mL of medium was added to the sediment. The cells were gently suspended in a pasteur pipette with a roasted tip until the cell mass disappeared, and then filtered through a 70 μm nylon cell strainer (Falcon). Cell culture was performed on 8-well chamber slides (Falcon). The day before, the coating [Poly-D-lysine (PDL, Sigma-Aldrich) was diluted with PBS to 0.005 mg / mL, sprinkled, and incubated at 37 ° C. Wash twice with sterile purified water on the day of culture. ], The cells were seeded. Culturing was started at 37 ° C., 10% CO 2 , saturated steam, and after 5 hours, the whole medium was replaced with a new medium containing B-27 supplement (Invitrogen) instead of horse serum. After conditioning medium or drug treatment, immunostaining was performed in the evaluation of axonal extension.
免疫染色は、薬物処置期間終了後、培地を除いてPBSで洗浄し、4%パラホルムアルデヒド(PFA,和光純薬)−PBS溶液を加え、2時間室温で静置し、細胞を固定した。0.3%トリトン(triton)X−100(和光純薬)−PBS溶液で5分間の洗浄を2回行った。その後、1次抗体溶液[0.3%トリトンX−100−PBS溶液、 1%正常ヤギ血清(normal goat serum、NGS、和光純薬)、1次抗体]を加え、4℃で一晩反応させた。 For immunostaining, after the end of the drug treatment period, the medium was removed, the cells were washed with PBS, 4% paraformaldehyde (PFA, Wako Pure Drug) -PBS solution was added, and the cells were allowed to stand at room temperature for 2 hours to fix the cells. Washing was performed twice for 5 minutes with 0.3% triton X-100 (Wako Pure Drug) -PBS solution. Then, the primary antibody solution [0.3% Triton X-100-PBS solution, 1% normal goat serum (NGS, Wako Pure Drug), primary antibody] was added, and the mixture was reacted overnight at 4 ° C. It was.
1次抗体には、マウス抗リン酸化型NF−Hモノクローナル抗体(1:250)(COVANCE, Emeryville, CA, USA)、ウサギ抗MAP2ポリクローナル抗体(1:2000)(abcam, Cambridge、英国)を用いた。翌日、1次抗体溶液を除き、0.3%トリトンX−100−PBS溶液で5分間の洗浄を2回行った後、2次抗体溶液[0.3%トリトンX−100−PBS溶液,Alexa Fluor 594標識ヤギ抗マウスIgG抗体(1:200)(Molecular Probes, Eugene, OR, USA)、Alexa Fluor 488 標識 ヤギ抗ウサギIgG抗体(1:200)(Molecular Probes, Eugene, OR, USA)]を加え、遮光下の室温で2時間反応させた。反応後、2次抗体溶液を除き、PBSで5分間、2回洗浄し、DAPI染色で核を染めた後、Aqua Poly Mount(Polysciences, Warrington, USA)で封入した。 As the primary antibody, mouse anti-phosphorylated NF-H monoclonal antibody (1: 250) (COVANCE, Emeryville, CA, USA) and rabbit anti-MAP2 polyclonal antibody (1: 2000) (abcam, Cambridge, UK) were used. There was. The next day, the primary antibody solution was removed, and after washing twice with 0.3% Triton X-100-PBS solution for 5 minutes, the secondary antibody solution [0.3% Triton X-100-PBS solution, Alexa Fluor 594-labeled goat anti-mouse IgG antibody (1: 200) (Molecular Probes, Eugene, OR, USA), Alexa Fluor 488-labeled goat anti-rabbit IgG antibody (1: 200) (Molecular Probes, Eugene, OR, USA)] In addition, the reaction was carried out at room temperature under shading for 2 hours. After the reaction, the secondary antibody solution was removed, the cells were washed twice with PBS for 5 minutes, the nuclei were stained with DAPI staining, and then encapsulated with Aqua Poly Mount (Polysciences, Warrington, USA).
蛍光顕微鏡(Cell Observer、Carl Zeiss、独国)を用いて、各薬物処置を施したウェル(well)から倍率10倍−20倍の対物レンズで画像を取得した。それらの画像について、画像解析ソフトMetaMorph(モレキュラーデバイス,東京)を用いて、画面全体のpNF−H陽性の軸索の長さと、各画像内のMAP2陽性神経細胞を計測し、1神経細胞あたりの軸索の長さを算出した。 Images were acquired from wells treated with each drug using a fluorescence microscope (Cell Observer, Carl Zeiss, Germany) with a 10-20x objective lens. For those images, the length of pNF-H-positive axons on the entire screen and the MAP2-positive nerve cells in each image were measured using the image analysis software MetaMorph (Molecular Device, Tokyo), and per nerve cell. The length of the axon was calculated.
<脊髄損傷(SCI)マウス:圧挫モデル>
8−10週齢の雌のddYマウスに挫傷を施した。すなわち、常法に従ってマウスの第8〜9腰椎を切除し、第11〜12胸髄へ定位固定装置(ナリシゲ社製)を用いて高さ3cmから6.5gの重りを一度落下させることにより挫傷を与えた。その後、常法に従って縫合などの外科的処置を施した。以下、このマウスをSCI圧挫モデルと称する。なおマウスは、餌と水を自由に摂取できる状態で、一定の環境条件(22±2℃、50±5%湿度、午前7時から開始される12時間の明暗サイクル)下で維持した。<Spinal Cord Injury (SCI) Mouse: Contusion Model>
8-10 week old female ddY mice were bruised. That is, the 8th to 9th lumbar vertebrae of the mouse were excised according to a conventional method, and a weight of 3 cm to 6.5 g in height was once dropped onto the 11th to 12th thoracic spines using a stereotaxic fixation device (manufactured by Narishige) to cause contusion. Gave. After that, surgical procedures such as suturing were performed according to a conventional method. Hereinafter, this mouse is referred to as an SCI crush model. Mice were maintained under constant environmental conditions (22 ± 2 ° C., 50 ± 5% humidity, 12-hour light-dark cycle starting at 7:00 am) in a state where food and water could be freely ingested.
<脊髄損傷(SCI)マウス:完全切断モデル>
8週齢の雌のddYマウスの第10腰椎を切除し、第1腰髄を露出させマイクロシーザーズで完全に垂直切断した。その後、常法に従って縫合などの外科的処置を施した。以下、このマウスをSCI完全切断モデルと称する。なおマウスは、餌と水を自由に摂取できる状態で、一定の環境条件(22±2℃、50±5%湿度、午前7時から開始される12時間の明暗サイクル)下で維持した。<Spinal cord injury (SCI) mouse: complete amputation model>
The 10th lumbar vertebrae of 8-week-old female ddY mice were excised to expose the 1st lumbar spine and completely vertically cut with Micro Caesars. After that, surgical procedures such as suturing were performed according to a conventional method. Hereinafter, this mouse is referred to as a SCI complete cleavage model. Mice were maintained under constant environmental conditions (22 ± 2 ° C., 50 ± 5% humidity, 12-hour light-dark cycle starting at 7:00 am) in a state where food and water could be freely ingested.
<TMSによる後肢運動機能の評価>
エキス等の投与後のマウスを個別にオープンフィールド(42cm×48cm×15cm)に移動させ、5分間観察し、後肢運動機能を評価した。脊髄損傷のモデル試験における後肢の運動機能を評価する基準として一般的に用いられるバッソマウススケール(Basso Mouse Scale:BMS、Engesser-Cesar C, Anderson AJ, Basso DM et al. (2005).)と、試験の精度を高めるために本発明者らがBMSに改変を加えた0〜30ポイントのトヤママウススケール(Toyama Mouse Scale、TMS)を用いて、オープンフィールドにおける移動行動を評価とした。<Evaluation of hind limb motor function by TMS>
Mice after administration of the extract and the like were individually moved to an open field (42 cm × 48 cm × 15 cm) and observed for 5 minutes to evaluate hindlimb motor function. Basso Mouse Scale (BMS, Engesser-Cesar C, Anderson AJ, Basso DM et al. (2005).), Which is commonly used as a criterion for evaluating the motor function of the hind limbs in a model test of spinal cord injury. In order to improve the accuracy of the test, we evaluated the movement behavior in the open field using the Toyama Mouse Scale (TMS) of 0 to 30 points, which was modified from BMS.
<統計処理>
本実施例において、得られた結果は以下の統計処理を行った:
一元配置分散分析(one-way ANOVA)、事後ダネット(Dunnett)検定、及び対応t−検定は、グラフパッド5(Graphpad Prism 6) (グラフパッドソフトウエア(Graphpad Software)社、ラホヤ(La Jolla)、カリフォルニア州、米国)を用いて行った。*P<0.05,**P<0.01,***P<0.001は統計学的に有意とし、平均値は標準誤差とともに示す。<Statistical processing>
In this example, the results obtained were subjected to the following statistical processing:
One-way ANOVA, post-Dunnett test, and corresponding t-test are available at Graphpad Prism 6 (Graphpad Software, La Jolla, La Jolla, Inc.) California, USA). * P <0.05, ** P <0.01, *** P <0.001 are statistically significant, and the average value is shown together with the standard error.
1.軸索伸展作用
(1−1)初代培養マウス骨格筋細胞を薬物処置して得た、培養上清(conditioned medium)を、初代培養マウス大脳皮質神経細胞に処置し、軸索伸展作用を調べた結果を図1に示す。なお、図中の白カラムは溶媒(滅菌水)を骨格筋細胞に処置して得たconditioned mediumを神経細胞に処置したものである。 1. 1. Axon extension action (1-1) The culture supernatant (conditioned medium) obtained by drug-treating primary cultured mouse skeletal muscle cells was treated with primary cultured mouse cerebral cortex neurons, and the axon extension action was examined. The results are shown in FIG. The white column in the figure is a conditioned medium obtained by treating skeletal muscle cells with a solvent (sterilized water) and treating nerve cells.
図1のAは、ニクジュウヨウエキス(培地中の最終濃度:10μg/mL、100μg/mL)を骨格筋細胞に処置した結果であり、そのconditioned mediumによって、ニクジュウヨウエキス処置群において有意に軸索密度が増加したことがわかる。 A in FIG. 1 shows the results of treating skeletal muscle cells with Nikujuyo extract (final concentration in medium: 10 μg / mL, 100 μg / mL), and the conditioned medium significantly axons in the Nikujuyo extract treatment group. It can be seen that the density has increased.
また、図1のBは、アクテオシド(培地中の最終濃度:1nM、100nM、1000nM)を骨格筋細胞に処置した結果であり、そのconditioned mediumによって有意に軸索密度が増加したことがわかる。 In addition, B in FIG. 1 shows the results of treating skeletal muscle cells with acteosides (final concentrations in the medium: 1 nM, 100 nM, 1000 nM), and it can be seen that the axon density was significantly increased by the conditioned medium.
(1−2)初代培養マウス大脳皮質神経細胞に、直接、薬物を処置し、軸索伸展作用を調べた結果を図2に示す。なお、図中の白カラムは薬物の代わりに溶媒(滅菌水)を培地に加えて処置した。 (1-2) Fig. 2 shows the results of directly treating primary cultured mouse cerebral cortical neurons with a drug and examining the axon extension action. The white column in the figure was treated by adding a solvent (sterilized water) to the medium instead of the drug.
図2のAは、ニクジュウヨウエキス(培地中の最終濃度:10μg/mL)を神経細胞に処置した結果であり、軸索伸展増加傾向が認められた。 FIG. 2A shows the results of treatment of nerve cells with Nikujuyo extract (final concentration in medium: 10 μg / mL), and an increasing tendency of axon extension was observed.
図2Bは、アクテオシド(培地中の最終濃度:1μM)を神経細胞に処置した結果であり、有意な軸索伸展作用が認められた。 FIG. 2B shows the results of treatment of nerve cells with acteoside (final concentration in medium: 1 μM), and a significant axon extension effect was observed.
2.筋細胞の増殖
初代培養マウス骨格筋細胞に薬物を処置し、筋細胞の増殖作用を調べた結果を図3に示す。 2. Proliferation of muscle cells FIG. 3 shows the results of treating the skeletal muscle cells of the primary cultured mouse with a drug and examining the proliferative effect of the muscle cells.
図3の結果から明らかなように、ニクジュウヨウエキス(培地中の最終濃度:1、100、100μg/mL)処置、アクテオシド(培地中の最終濃度:1、100、1000nM)処置いずれにおいても、骨格筋細胞の増殖が有意に促進した。なお、IGF−1は陽性コントロールとして用いた。 As is clear from the results of FIG. 3, the skeletal skeletal structure was treated with Nikujuyo extract (final concentration in medium: 1,100,100 μg / mL) and acteoside (final concentration in medium: 1,100,1000 nM). The proliferation of muscle cells was significantly promoted. IGF-1 was used as a positive control.
3.脊髄損傷により低下した慢性期における運動機能の改善
(3−1)マウスに圧挫による脊髄損傷を与え、45日間経過後、いわゆる受傷後慢性期になってから後肢大腿二頭筋にニクジュウヨウエキスを筋肉注射し(2.0mg/肢、左右後肢へ筋肉注射、3回/週)、35日間にわたる薬物投与期間中の後肢運動機能の推移を調べた。結果を図4に示す。 3. 3. Improvement of motor function in the chronic phase decreased due to spinal cord injury (3-1) After 45 days have passed since the mouse was injured by spinal cord injury and the so-called post-injury chronic phase was reached, the biceps femoris extract was applied to the hind limb biceps femoris. Was intramuscularly injected (2.0 mg / limb, intramuscular injection into the left and right hind limbs, 3 times / week), and the transition of hind limb motor function during the drug administration period over 35 days was examined. The results are shown in FIG.
図4Aは、バッソマウススケール(Basso Mouse Scale、BMS)で評価した結果であり、ニクジュウヨウエキス注射群で運動機能が改善する傾向が示された。 FIG. 4A shows the results of evaluation on the Basso Mouse Scale (BMS), and showed a tendency for the motor function to improve in the Nikujuyo extract injection group.
図4Bは、トヤママウススケール(Toyama Mouse Scale、TMS)で評価した結果であり、ニクジュウヨウエキス投与群が有意に後肢運動機能の改善を示した。 FIG. 4B shows the results of evaluation on the Toyama Mouse Scale (TMS), and the Nikujuyo extract-administered group showed a significant improvement in hindlimb motor function.
(3−2)マウスに圧挫による脊髄損傷を与え、45日間経過後、いわゆる受傷後慢性期になってから後肢大腿二頭筋にアクテオシドを筋肉注射し(0.1mg/肢、左右後肢へ筋肉注射、3回/週)、35日間にわたる薬物投与期間中の後肢運動機能の推移を調べた。結果を図5に示す。 (3-2) Mice were injured by spinal cord injury due to crush, and after 45 days had passed, after the so-called chronic phase after injury, acteoside was intramuscularly injected into the biceps femoris of the hind limbs (0.1 mg / limb, to the left and right hind limbs). The transition of hindlimb motor function during the drug administration period of 35 days with intramuscular injection (3 times / week) was examined. The results are shown in FIG.
図5AはBMS、図5BはTMSで評価した結果であり、いずれにおいても、アクテオシド注射群で有意な後肢運動機能改善効果が認められた。 FIG. 5A shows the results evaluated by BMS and FIG. 5B shows the results evaluated by TMS, and in both cases, a significant hindlimb motor function improving effect was observed in the acteoside injection group.
(3−3)マウスに完全脊髄切断による損傷を与え、35日間経過後、いわゆる受傷後慢性期になってから後肢大腿二頭筋にニクジュウヨウエキスを筋肉注射し(2.0mg/肢、左右後肢へ筋肉注射、3回/週)、56日間にわたる薬物投与期間中の後肢運動機能の推移を調べた。結果を図6に示す。 (3-3) Mice were injured by complete spinal cord transection, and 35 days later, after the so-called chronic phase after injury, Nikujuyo extract was intramuscularly injected into the biceps femoris of the hind limbs (2.0 mg / limb, left and right). Intramuscular injection into the hind limbs (3 times / week), the transition of hind limb motor function during the drug administration period over 56 days was examined. The results are shown in FIG.
図6AはBMS、図6BはTMSで評価した結果であり、いずれにおいても、ニクジュウヨウエキス注射群で有意な後肢運動機能改善効果が認められた。 FIG. 6A shows the results evaluated by BMS and FIG. 6B shows the results evaluated by TMS, and in both cases, a significant hindlimb motor function improving effect was observed in the Nikujuyo extract injection group.
4.筋重量の増加
マウスに完全脊髄切断による損傷を与え、35日間経過後、いわゆる受傷後慢性期になってから後肢大腿二頭筋にニクジュウヨウエキスを筋肉注射し(2.0mg/肢、左右後肢へ筋肉注射、3回/週)、56日間にわたる薬物投与後の腓腹筋の湿重量を調べた。結果を図7に示す。 4. Increased muscle weight Mice were injured by complete spinal cord transection, and 35 days later, after the so-called chronic phase after injury, Nikujuyo extract was intramuscularly injected into the biceps femoris of the hind limbs (2.0 mg / limb, left and right hind limbs). Wet weight of the gastrocnemius muscle was examined after intramuscular injection, 3 times / week) and drug administration for 56 days. The results are shown in FIG.
図7の結果から明らかなように、未切断群と比べて、切断群の生理食塩水投与マウスでは著しく骨格筋重量が減少していた。しかし、ニクジュウヨウエキス注射により、腓腹筋重量の増加が認められた。 As is clear from the results of FIG. 7, the skeletal muscle weight was significantly reduced in the saline-administered mice in the amputated group as compared with the uncut group. However, an increase in gastrocnemius muscle weight was observed by injection of Nikujuyo extract.
5.経口投与
(5−1)マウスに圧挫による脊髄損傷を与え、51日間経過後、いわゆる受傷後慢性期になってからニクジュウヨウエキスを経口投与し、脊髄損傷受傷後から薬物投与終了までの後肢運動機能の推移を調べた。なお、投与量は500mg/kg/日とし、138日間の投与を行った。結果を図8に示す。 5. Oral administration (5-1) Mice are injured by spinal cord injury due to crushing, and after 51 days, the so-called post-injury chronic phase is followed by oral administration of Nikujuyo extract, and the hind limbs from the time of spinal cord injury to the end of drug administration. The transition of motor function was investigated. The dose was 500 mg / kg / day, and administration was performed for 138 days. The results are shown in FIG.
図8AはBMS、図8BはTMSで評価した結果である。いずれにおいても、ニクジュウヨウエキス経口投与群で有意な後肢運動機能改善効果が認められた。 FIG. 8A is the result of evaluation by BMS, and FIG. 8B is the result of evaluation by TMS. In all cases, a significant hindlimb motor function improving effect was observed in the oral administration group of Nikujuyo extract.
(5−2)マウスに圧挫による脊髄損傷を与え、51日間経過後、いわゆる受傷後慢性期になってからニクジュウヨウエキスを胃ゾンデにて経口投与し、薬物投与終了後の後肢骨格筋(大腿二頭筋、前脛骨筋、外側広筋、腓腹筋)の湿重量を調べ、体重に対する割合(筋湿重量/体重)を算出した。なお、投与量は500mg/kg/日とし、138日間行った。
結果を図9に示す。図9において、Shamは、偽手術群、SCI/Vehは脊髄損傷群に溶媒投与、SCI/NJは脊髄損傷群にニクジュウヨウエキス投与を示す。溶媒は滅菌水であり、ニクジュウヨウエキスを滅菌水に5質量%を溶解したものを投与した。(5-2) Spinal cord injury due to crushing was given to the mouse, and 51 days later, after the so-called chronic phase after the injury, the biceps femoris extract was orally administered by gastrocnemius, and the tibialis anterior muscle after the drug administration was completed. The wet weight of the biceps femoris, tibialis anterior, vastus lateralis, and gastrocnemius muscles) was examined, and the ratio to body weight (muscle wet weight / body weight) was calculated. The dose was 500 mg / kg / day, and the dose was 138 days.
The results are shown in FIG. In FIG. 9, Sham indicates the sham surgery group, SCI / Veh indicates the solvent administration to the spinal cord injury group, and SCI / NJ indicates the Nikujuyo extract administration to the spinal cord injury group. The solvent was sterilized water, and a solution of Nikujuyo extract in sterilized water in an amount of 5% by mass was administered.
図9の結果から明らかなように、腓腹筋、前脛骨筋において、有意な筋重量増加が認められた。また、外側広筋、大腿二頭筋においてもニクジュウヨウエキス投与による筋重量増加傾向が見られた。 As is clear from the results of FIG. 9, a significant increase in muscle weight was observed in the gastrocnemius muscle and the tibialis anterior muscle. In addition, the vastus lateralis muscle and the biceps femoris also tended to increase muscle weight due to the administration of Nikujuyo extract.
6.ペリオスチン及びPKM2の分泌促進
初代培養マウス骨格筋細胞をアクテオシド処置して得たconditioned mediumに増加するタンパク質を同定した。結果を図10に示す。 6. We identified proteins that increase to the conditioned medium obtained by acteoside treatment of skeletal muscle cells of primary cultured mice that promote the secretion of periostin and PKM2 . The results are shown in FIG.
図10Aは、アクテオシドを骨格筋細胞に処置して得たconditioned mediumを電気泳動し、銀染色した結果である。2か所のバンドがアクテオシドの用量依存的に増加した。
このバンドをそれぞれゲルごと切り出し、ゲル内トリプシン消化の後、nano LC-MS/MS解析を実施した。ペプチド断片の質量パターンの結果より、2本のバンドは、ペリオスチンとPKM2である可能性が示された。FIG. 10A shows the results of electrophoresing and silver staining of a conditioned medium obtained by treating skeletal muscle cells with acteoside. Two bands increased in a dose-dependent manner with acteoside.
Each of these bands was excised together with the gel, and after in-gel trypsin digestion, nano LC-MS / MS analysis was performed. The results of the mass pattern of the peptide fragments indicated that the two bands could be periostin and PKM2.
図10Bは、アクテオシドを骨格筋細胞に処置して得たconditioned mediumをペリオスチンの抗体でウェスタンブロッティングした結果、図10Cは、アクテオシドを滅菌水に溶解し培地に加えて骨格筋細胞に処置して得たconditioned mediumをPKM2の抗体でウェスタンブロッティングした結果である。アクテオシドを骨格筋細胞に処置して得たconditioned medium中に、確かにペリオスチンとPKM2が増加していることが確認できた。 FIG. 10B shows the results of Western blotting of the conditioned medium obtained by treating skeletal muscle cells with acteoside with an antibody of periostin. FIG. 10C shows the result of dissolving acteoside in sterile water and adding it to the medium to treat skeletal muscle cells. This is the result of Western blotting of the conditioned medium with an antibody of PKM2. It was confirmed that periostin and PKM2 were certainly increased in the conditioned medium obtained by treating skeletal muscle cells with acteoside.
7.ペリオスチン・PKM2による機能
(7−1)初代培養マウス大脳皮質神経細胞にリコンビナントペリオスチン又はリコンビナントPKM2(いずれも市販品)処置し、軸索伸展作用を調べた。図11に結果を示す。 7. Function by periostin / PKM2 (7-1) Primary cultured mouse Cerebral cortex neurons were treated with recombinant periostin or recombinant PKM2 (both commercially available), and the axon extension effect was examined. The results are shown in FIG.
図11Aは、リコンビナントペリオスチンを滅菌水に溶解し培地に加えて神経細胞に処置した結果であり、軸索伸展作用が有意に見られた。 FIG. 11A shows the results of treating nerve cells by dissolving recombinant periostin in sterile water and adding it to a medium, and a significant axon extension effect was observed.
図11Bは、リコンビナントPKM2を滅菌水に溶解し培地に加えて神経細胞に処置した結果であり、10,100,1000ng/mlの用量において有意な軸索伸展作用が認められた。 FIG. 11B shows the results of treating the nerve cells by dissolving the recombinant PKM2 in sterile water and adding it to the medium, and a significant axon stretching effect was observed at a dose of 10,100,1000 ng / ml.
(7−2)初代培養マウス骨格筋細胞にリコンビナントペリオスチンまたはリコンビナントPKM2(いずれも市販品)を処置し、細胞増殖作用を調べた。結果を図12に示す。 (7-2) Primary cultured mouse skeletal muscle cells were treated with recombinant periostin or recombinant PKM2 (both commercially available), and the cell proliferation effect was examined. The results are shown in FIG.
図12Aは、リコンビナントペリオスチンを滅菌水に溶解し培地に加えて骨格筋細胞に処置した結果であり、有意に増殖が促進されていた。 FIG. 12A shows the results of treating skeletal muscle cells by dissolving recombinant periostin in sterile water and adding it to the medium, and the proliferation was significantly promoted.
図12Bは、リコンビナントPKM2を滅菌水に溶解し培地に加えて骨格筋細胞に処置した結果であり、有意に増殖が促進されていた。 FIG. 12B shows the results of treating the skeletal muscle cells by dissolving the recombinant PKM2 in sterile water and adding it to the medium, and the proliferation was significantly promoted.
8.アクテオシドによる慢性期脊髄損傷マウスの運動機能改善、筋萎縮改善および運動神経への前シナプス投射の増加
(8−1)マウスに圧挫による脊髄損傷を与え、30日間経過後、いわゆる受傷後慢性期になってからアクテオシドを筋肉注射し(0.2mg/肢、左右後肢へ筋肉注射、3回/週)、脊髄損傷受傷後から薬物投与終了までの後肢運動機能の推移を調べた。なお、投与期間は62日間とした。結果を図13に示す。 8. Chronic spinal cord injury due to acteoside Improvement of motor function, improvement of muscle atrophy and increase of presynaptic projection to motor nerves (8-1) Spinal cord injury due to crushing in mice, 30 days later, so-called post-injury chronic phase After that, acteoside was injected intramuscularly (0.2 mg / limb, intramuscular injection into the left and right hind limbs, 3 times / week), and the transition of hind limb motor function from the injury of SCI to the end of drug administration was investigated. The administration period was 62 days. The results are shown in FIG.
図13AはBMS、図13BはTMSで評価した結果である。いずれにおいても、アクテオシド筋肉注射群で有意な後肢運動機能改善効果が認められた。 FIG. 13A is the result of evaluation by BMS, and FIG. 13B is the result of evaluation by TMS. In all cases, a significant hindlimb motor function improving effect was observed in the acteoside intramuscular injection group.
(8−2)マウスに圧挫による脊髄損傷を与え、30日間経過後、いわゆる受傷後慢性期になってからアクテオシドを筋肉注射し(0.2mg/肢、左右後肢へ筋肉注射、3回/週)、薬物投与終了後の後肢骨格筋(大腿二頭筋、前脛骨筋、外側広筋、腓腹筋)の湿重量を調べ、体重に対する割合(筋湿重量/体重)を算出した。なお、投与期間は62日間とした。
結果を図14に示す。図14において、Shamは偽手術群、SCI/Vehは脊髄損傷群に溶媒投与、SCI/ACは脊髄損傷群にアクテオシド投与を示す。溶媒は生理食塩水とした。(8-2) Mice were injured by spinal cord injury due to crush, and 30 days later, after the so-called chronic phase after injury, acteoside was intramuscularly injected (0.2 mg / limb, intramuscular injection into the left and right hind limbs, 3 times / Week), the wet weight of the hindlimb skeletal muscles (biceps femoris, tibialis anterior muscle, vastus lateralis muscle, gastrocnemius muscle) after the end of drug administration was examined, and the ratio to the body weight (muscular wet weight / body weight) was calculated. The administration period was 62 days.
The results are shown in FIG. In FIG. 14, Sham indicates sham surgery group, SCI / Veh indicates solvent administration to the spinal cord injury group, and SCI / AC indicates acteoside administration to the spinal cord injury group. The solvent was physiological saline.
図14の結果から明らかなように、大腿二頭筋、前脛骨筋において、有意な筋重量増加が認められた。また、外側広筋、腓腹筋においてもアクテオシド投与による筋重量増加傾向が見られた。 As is clear from the results of FIG. 14, a significant increase in muscle weight was observed in the biceps femoris and tibialis anterior muscles. In addition, the vastus lateralis muscle and gastrocnemius muscle also tended to increase muscle weight due to the administration of acteoside.
(8−3)マウスに圧挫による脊髄損傷を与え、30日間経過後、いわゆる受傷後慢性期になってからアクテオシドを筋肉注射し(0.2mg/肢、左右後肢へ筋肉注射、3回/週)、薬物投与終了後の脊髄組織のうち大腿二頭筋、前脛骨筋、外側広筋、腓腹筋へ投射する運動神経が局在する腰髄部を摘出した。14μm厚の連続矢状断切片を作製し、運動神経マーカーのcholine acetyltransferaseに対する抗体、前シナプスマーカーのsynaptophysinに対する抗体による免疫染色を行った。運動神経細胞上に投射する前シナプスの面積を定量した。結果を図15に示す。図15において、Shamは偽手術群、SCI/Vehは脊髄損傷群に溶媒投与、SCI/ACは脊髄損傷群にアクテオシド投与を示す。溶媒は生理食塩水とした。 (8-3) Mice were injured by spinal cord injury due to crush, and after 30 days, acteoside was intramuscularly injected (0.2 mg / limb, intramuscular injection into left and right hind limbs, 3 times /) in the so-called chronic phase after injury. Week), the lumbar spinal cord was removed from the spinal cord tissue after drug administration, where the motor nerves projecting to the biceps femoris, tibialis anterior, vastus lateralis, and gastrocnemius muscles were localized. A 14 μm-thick continuous sagittal section was prepared and immunostained with an antibody against the motor nerve marker choline acetyltransferase and an antibody against the presynaptic marker synaptophysin. The area of presynapses projected onto motor neurons was quantified. The results are shown in FIG. In FIG. 15, Sham indicates sham surgery group, SCI / Veh indicates solvent administration to the spinal cord injury group, and SCI / AC indicates acteoside administration to the spinal cord injury group. The solvent was physiological saline.
図15の結果から明らかなように、アクテオシド投与により、運動神経細胞へ投射する前シナプスが増加した。 As is clear from the results of FIG. 15, administration of acteoside increased presynapses that project to motor neurons.
9.ニクジュウヨウエキスによる筋萎縮マウスの運動機能改善
(9−1)マウスの両後肢の関節を伸展位で固定するキャストを装着し、12日後にキャストを外した。これは、キャスト未装着群(偽手術群)と比較してキャスト装着群では有意な骨格筋の萎縮が認められる、サルコペニアのマウスモデルである。キャスト装着期間中、ニクジュウヨウエキスを毎日経口投与した。投与量は50mg/kg/日、および100mg/kg/日とした。キャストを外した翌日、ビームテストによる運動機能の評価を行った。結果(ニクジュウヨウエキスを滅菌水に溶解し、筋萎縮マウスに経口投与した13日目の運動機能の結果)を図16に示す。ビームテストは幅5mm、9mm、12mmのバーの上をマウスが歩行する様子を観察し、左右の後肢が滑り落ちる頻度をスコア化して総得点を算出するもので、点数が高いほど失敗なく歩行できることを示す。 9. Improvement of motor function in muscularly atrophied mice by Nikujuyo extract (9-1) A cast was attached to fix the joints of both hind limbs of the mouse in the extended position, and the cast was removed 12 days later. This is a mouse model of sarcopenia in which significant skeletal muscle atrophy is observed in the cast-attached group compared to the non-cast-attached group (sham-operated group). During the cast wearing period, Nikujuyo extract was orally administered daily. The dose was 50 mg / kg / day and 100 mg / kg / day. The day after the cast was removed, the motor function was evaluated by a beam test. The results (results of motor function on the 13th day when the Nikujuyo extract was dissolved in sterile water and orally administered to muscular atrophied mice) are shown in FIG. The beam test observes the mouse walking on bars with widths of 5 mm, 9 mm, and 12 mm, scores the frequency with which the left and right hind limbs slide down, and calculates the total score.The higher the score, the more you can walk without failure. Shown.
図16の結果から明らかなように、ニクジュウヨウエキスにより、筋萎縮マウスの運動機能に、有意な改善作用が見られた。 As is clear from the results of FIG. 16, the Nikujuyo extract had a significant improving effect on the motor function of the muscular atrophied mice.
本発明では、筋細胞の増殖、筋肉の増加や、ペリオスチンやPKM2の分泌に有用な剤又は組成物等を提供できる。 INDUSTRIAL APPLICABILITY The present invention can provide an agent or composition useful for muscle cell proliferation, muscle gain, and secretion of periostin and PKM2.
Claims (13)
(1)神経疾患の治療及び/又は改善
(2)筋疾患の治療及び/又は改善
(3)サルコペニアの治療及び/又は改善
(4)ロコモーティブシンドロームの治療及び/又は改善
(5)麻痺の治療及び/又は改善
(6)運動機能及び/又は体力の改善及び/又は維持The agent or composition according to any one of claims 1 to 5, for use in at least one application selected from the following (1) to (6).
(1) Treatment and / or improvement of neurological disorders (2) Treatment and / or improvement of muscle disorders (3) Treatment and / or improvement of sarcopenia (4) Treatment and / or improvement of locomotive syndrome (5) Treatment and / or improvement of paralysis / Or improvement (6) Improvement and / or maintenance of motor function and / or physical strength
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2017201435 | 2017-10-17 | ||
JP2017201435 | 2017-10-17 | ||
PCT/JP2018/025119 WO2019077810A1 (en) | 2017-10-17 | 2018-07-02 | Periostin and pkm2 secretion promoter |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2019077810A1 true JPWO2019077810A1 (en) | 2020-11-05 |
JP7356710B2 JP7356710B2 (en) | 2023-10-05 |
Family
ID=66173931
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2019549111A Active JP7356710B2 (en) | 2017-10-17 | 2018-07-02 | Periostin and PKM2 secretion promoter |
Country Status (2)
Country | Link |
---|---|
JP (1) | JP7356710B2 (en) |
WO (1) | WO2019077810A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112587487A (en) * | 2020-11-30 | 2021-04-02 | 建昌帮药业有限公司 | Echinacoside orally disintegrating tablet and preparation method thereof |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100166827A1 (en) * | 2007-01-22 | 2010-07-01 | Bernhard Kuhn | Periostin induces proliferation of cardiomyocytes and promotes cardiac regeneration |
US20110144040A1 (en) * | 2009-12-10 | 2011-06-16 | MN Intellectual Property Corp, LLC | Acteoside and acteoside-rich plant extracts for increasing athletic performance in humans |
JP2016521717A (en) * | 2013-06-13 | 2016-07-25 | ナタク バイオテク、エセ. エレNatac Biotech,S.L | Combinations of pentacyclic triterpenes with hydroxytyrosol and its derivatives |
WO2017121333A1 (en) * | 2016-01-12 | 2017-07-20 | 杏辉天力(杭州)药业有限公司 | Use of cistanche tubulosa extract and isoacteoside in protection of muscles |
-
2018
- 2018-07-02 JP JP2019549111A patent/JP7356710B2/en active Active
- 2018-07-02 WO PCT/JP2018/025119 patent/WO2019077810A1/en active Application Filing
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100166827A1 (en) * | 2007-01-22 | 2010-07-01 | Bernhard Kuhn | Periostin induces proliferation of cardiomyocytes and promotes cardiac regeneration |
US20110144040A1 (en) * | 2009-12-10 | 2011-06-16 | MN Intellectual Property Corp, LLC | Acteoside and acteoside-rich plant extracts for increasing athletic performance in humans |
JP2016521717A (en) * | 2013-06-13 | 2016-07-25 | ナタク バイオテク、エセ. エレNatac Biotech,S.L | Combinations of pentacyclic triterpenes with hydroxytyrosol and its derivatives |
WO2017121333A1 (en) * | 2016-01-12 | 2017-07-20 | 杏辉天力(杭州)药业有限公司 | Use of cistanche tubulosa extract and isoacteoside in protection of muscles |
Non-Patent Citations (5)
Title |
---|
BRAIN RESEARCH, vol. 1346, JPN6023003114, 2010, pages 224 - 236, ISSN: 0005118559 * |
DEVELOPMENT,GROWTH & DIFFERENTIATION, vol. 57, no. 3, JPN6022022750, 2015, pages 200 - 208, ISSN: 0005118557 * |
FOOD STYLE 21, vol. 12, no. 7, JPN6022022747, 2008, pages 21 - 23, ISSN: 0005118555 * |
JOURNAL OF NEUROSCIENCE, vol. 34, no. 7, JPN6022022751, 2014, pages 2438 - 2443, ISSN: 0005118558 * |
NAUNYN-SCHMIEDEBERG'S ARCHIVES OF PHARMACOLOGY, vol. 382, no. 4, JPN6022022748, 2010, pages 331 - 345, ISSN: 0005118556 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112587487A (en) * | 2020-11-30 | 2021-04-02 | 建昌帮药业有限公司 | Echinacoside orally disintegrating tablet and preparation method thereof |
Also Published As
Publication number | Publication date |
---|---|
JP7356710B2 (en) | 2023-10-05 |
WO2019077810A1 (en) | 2019-04-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7379152B2 (en) | Composition for inhibiting muscle fibrosis | |
JP2015523333A (en) | Cosmetic, pharmaceutical and food composition containing a crushed or extracted fish eyeball | |
US20220193147A1 (en) | Anti-aging Composition Containing Akkermansia Muciniphila as Active Ingredient and a Method for Preventing Aging Using Thereof | |
KR102175468B1 (en) | Cosmetic composition for promoting a differentiation of adipocyte comprising hydroxy pyranone derivative | |
KR102091620B1 (en) | Pharmaceutical composition for prevention or treatment of blood brain barrier disorder comprising l-serine as an effective component and health functional food comprising the same | |
CN107530391A (en) | Cooperative compositions for osteoarthritis | |
KR101746388B1 (en) | Phamaceutial composition comprising of dichloromethane fraction of Aster yomena for immune enhancing | |
JP2016027031A (en) | Medicine or food for neural network reconstruction/activation | |
JP7356710B2 (en) | Periostin and PKM2 secretion promoter | |
JP2008266156A (en) | Heparin-binding epidermal growth factor-like growth factor gene expression promoter containing plant extract | |
EP2765869B1 (en) | Egg preparation with regenerating, analgesic and/or anti-inflammatory properties | |
KR101636345B1 (en) | Composition for preventing or treating thyroid disorders comprising aloe extracts or fraction thereof | |
KR100945960B1 (en) | A composition containing Arazyme for the prevention and treatment of arthritis | |
JP6360357B2 (en) | Method for inhibiting adipocyte formation | |
KR20170055614A (en) | A composition for preventing or treating menopausal cardiovascular disease comprising Cuscuta japonica Chois extract | |
KR20200022789A (en) | Pharmaceutical composition for preventing or treating muscle weakness diseases comprising deoxycholic acid or pharmaceutically acceptable salt thereof | |
JP2004083416A (en) | Skin care preparation for external use and food and drink product | |
JP7382032B2 (en) | Composition for preventing or treating rheumatoid arthritis containing snake venom | |
WO2017030167A1 (en) | Agent for reconstructing/activating neural network | |
US11679134B2 (en) | Pharmaceutical composition, food composition and food additive for preventing, alleviating or treating muscle loss, weakness and atrophy, containing, as active ingredient, Enterococcus faecalis, culture liquid thereof or dead cells thereof | |
KR20190136543A (en) | Compostion for preventing or treating the Developmental Disability comprising Humulus japonicus extract as active ingredient | |
JP6547183B2 (en) | Composition for prevention or treatment of osteoarthritis comprising triterpene as an active ingredient | |
JP2020094015A (en) | Hyaluronic acid production promoter, joint function improving agent, and oral composition | |
JP7319611B2 (en) | Proliferation promoter for stem cells | |
KR101606645B1 (en) | Composition for Preventing and Treating Immune diseases Comprising Daurinol |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A80 | Written request to apply exceptions to lack of novelty of invention |
Free format text: JAPANESE INTERMEDIATE CODE: A80 Effective date: 20200305 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200408 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20210628 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220607 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220804 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20221004 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20221201 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230131 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230327 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230509 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230627 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230801 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230822 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20230905 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20230915 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7356710 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |