CN104055759B - A kind of pharmaceutical composition and application thereof - Google Patents
A kind of pharmaceutical composition and application thereof Download PDFInfo
- Publication number
- CN104055759B CN104055759B CN201410267131.6A CN201410267131A CN104055759B CN 104055759 B CN104055759 B CN 104055759B CN 201410267131 A CN201410267131 A CN 201410267131A CN 104055759 B CN104055759 B CN 104055759B
- Authority
- CN
- China
- Prior art keywords
- pharmaceutical composition
- rat
- present
- medicine
- group
- 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.)
- Expired - Fee Related
Links
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 65
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 claims abstract description 84
- 208000006096 Attention Deficit Disorder with Hyperactivity Diseases 0.000 claims abstract description 72
- 208000036864 Attention deficit/hyperactivity disease Diseases 0.000 claims abstract description 72
- 229960002430 atomoxetine Drugs 0.000 claims abstract description 63
- 239000003814 drug Substances 0.000 claims abstract description 47
- 210000004556 brain Anatomy 0.000 claims abstract description 42
- 229960003080 taurine Drugs 0.000 claims abstract description 41
- VHGCDTVCOLNTBX-QGZVFWFLSA-N atomoxetine Chemical compound O([C@H](CCNC)C=1C=CC=CC=1)C1=CC=CC=C1C VHGCDTVCOLNTBX-QGZVFWFLSA-N 0.000 claims abstract 6
- 238000002360 preparation method Methods 0.000 claims description 11
- 239000000203 mixture Substances 0.000 claims description 10
- 229940079593 drug Drugs 0.000 claims description 8
- 239000002671 adjuvant Substances 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 3
- 239000003112 inhibitor Substances 0.000 claims description 3
- 239000000314 lubricant Substances 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 3
- 239000012190 activator Substances 0.000 claims description 2
- 230000002490 cerebral effect Effects 0.000 abstract description 24
- 101150078861 fos gene Proteins 0.000 abstract description 24
- 238000002474 experimental method Methods 0.000 abstract description 22
- 230000000366 juvenile effect Effects 0.000 abstract description 22
- 230000000694 effects Effects 0.000 abstract description 19
- 238000011552 rat model Methods 0.000 abstract description 10
- 210000002442 prefrontal cortex Anatomy 0.000 abstract description 6
- 230000002195 synergetic effect Effects 0.000 abstract description 5
- 230000003078 antioxidant effect Effects 0.000 abstract description 3
- 229940000406 drug candidate Drugs 0.000 abstract 1
- 241000700159 Rattus Species 0.000 description 85
- LUCXVPAZUDVVBT-UNTBIKODSA-N atomoxetine hydrochloride Chemical compound Cl.O([C@H](CCNC)C=1C=CC=CC=1)C1=CC=CC=C1C LUCXVPAZUDVVBT-UNTBIKODSA-N 0.000 description 60
- 238000012360 testing method Methods 0.000 description 30
- 210000001519 tissue Anatomy 0.000 description 29
- 239000013641 positive control Substances 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 108010015720 Dopamine beta-Hydroxylase Proteins 0.000 description 15
- 102100033156 Dopamine beta-hydroxylase Human genes 0.000 description 15
- DIVDFFZHCJEHGG-UHFFFAOYSA-N oxidopamine Chemical compound NCCC1=CC(O)=C(O)C=C1O DIVDFFZHCJEHGG-UHFFFAOYSA-N 0.000 description 13
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 12
- 229960003180 glutathione Drugs 0.000 description 12
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical group [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 12
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 12
- 102000019197 Superoxide Dismutase Human genes 0.000 description 11
- 108010012715 Superoxide dismutase Proteins 0.000 description 11
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 101100120538 Rattus norvegicus Fos gene Proteins 0.000 description 10
- 239000012153 distilled water Substances 0.000 description 10
- 230000037396 body weight Effects 0.000 description 9
- 210000002569 neuron Anatomy 0.000 description 9
- 229960002748 norepinephrine Drugs 0.000 description 9
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 230000006399 behavior Effects 0.000 description 7
- 230000001660 hyperkinetic effect Effects 0.000 description 7
- 239000013642 negative control Substances 0.000 description 7
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 6
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 6
- 101100028791 Caenorhabditis elegans pbs-5 gene Proteins 0.000 description 6
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical group OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 6
- 229930195725 Mannitol Natural products 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 229920002472 Starch Polymers 0.000 description 6
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 235000019359 magnesium stearate Nutrition 0.000 description 6
- 239000000594 mannitol Substances 0.000 description 6
- 235000010355 mannitol Nutrition 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 239000000741 silica gel Substances 0.000 description 6
- 229910002027 silica gel Inorganic materials 0.000 description 6
- 235000019698 starch Nutrition 0.000 description 6
- 239000008107 starch Substances 0.000 description 6
- 101100499815 Rattus norvegicus Dbh gene Proteins 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 210000005064 dopaminergic neuron Anatomy 0.000 description 5
- 239000000890 drug combination Substances 0.000 description 5
- 238000012346 open field test Methods 0.000 description 5
- 210000002381 plasma Anatomy 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 101000606117 Rattus norvegicus Tyrosine 3-monooxygenase Proteins 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 230000036772 blood pressure Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 230000003203 everyday effect Effects 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 239000012188 paraffin wax Substances 0.000 description 4
- 230000001681 protective effect Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 102000012440 Acetylcholinesterase Human genes 0.000 description 3
- 108010022752 Acetylcholinesterase Proteins 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 229940022698 acetylcholinesterase Drugs 0.000 description 3
- 230000037005 anaesthesia Effects 0.000 description 3
- 229960002828 atomoxetine hydrochloride Drugs 0.000 description 3
- 230000006287 biotinylation Effects 0.000 description 3
- 238000007413 biotinylation Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 3
- 229960002327 chloral hydrate Drugs 0.000 description 3
- 230000005611 electricity Effects 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 238000003304 gavage Methods 0.000 description 3
- 238000001802 infusion Methods 0.000 description 3
- 230000036542 oxidative stress Effects 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000007619 statistical method Methods 0.000 description 3
- 239000003826 tablet Substances 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 230000000007 visual effect Effects 0.000 description 3
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 241001062009 Indigofera Species 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 101000959274 Tenebrio molitor Antidiuretic factor A Proteins 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000003064 anti-oxidating effect Effects 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000000747 cardiac effect Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- -1 decoction Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 210000001652 frontal lobe Anatomy 0.000 description 2
- 238000013115 immunohistochemical detection Methods 0.000 description 2
- 230000002055 immunohistochemical effect Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000000415 inactivating effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 230000002474 noradrenergic effect Effects 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 230000010412 perfusion Effects 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 230000003518 presynaptic effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108700017511 rat Y acceptor Proteins 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 230000000862 serotonergic effect Effects 0.000 description 2
- 230000002269 spontaneous effect Effects 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 210000000115 thoracic cavity Anatomy 0.000 description 2
- 238000011706 wistar kyoto rat Methods 0.000 description 2
- KWTSXDURSIMDCE-QMMMGPOBSA-N (S)-amphetamine Chemical compound C[C@H](N)CC1=CC=CC=C1 KWTSXDURSIMDCE-QMMMGPOBSA-N 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- 101100426754 Caenorhabditis elegans try-5 gene Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- HCYAFALTSJYZDH-UHFFFAOYSA-N Desimpramine Chemical compound C1CC2=CC=CC=C2N(CCCNC)C2=CC=CC=C21 HCYAFALTSJYZDH-UHFFFAOYSA-N 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 206010020651 Hyperkinesia Diseases 0.000 description 1
- 208000000269 Hyperkinesis Diseases 0.000 description 1
- 206010021403 Illusion Diseases 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 1
- 241001417092 Macrouridae Species 0.000 description 1
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 1
- DUGOZIWVEXMGBE-UHFFFAOYSA-N Methylphenidate Chemical compound C=1C=CC=CC=1C(C(=O)OC)C1CCCCN1 DUGOZIWVEXMGBE-UHFFFAOYSA-N 0.000 description 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 description 1
- 102000008109 Mixed Function Oxygenases Human genes 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 102000008092 Norepinephrine Plasma Membrane Transport Proteins Human genes 0.000 description 1
- 108010049586 Norepinephrine Plasma Membrane Transport Proteins Proteins 0.000 description 1
- 208000028017 Psychotic disease Diseases 0.000 description 1
- 235000009233 Stachytarpheta cayennensis Nutrition 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 101710164184 Synaptic vesicular amine transporter Proteins 0.000 description 1
- 102100034333 Synaptic vesicular amine transporter Human genes 0.000 description 1
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 238000001949 anaesthesia Methods 0.000 description 1
- 239000002269 analeptic agent Substances 0.000 description 1
- 230000003556 anti-epileptic effect Effects 0.000 description 1
- 239000001961 anticonvulsive agent Substances 0.000 description 1
- 229960003965 antiepileptics Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 229940076134 benzene Drugs 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000008499 blood brain barrier function Effects 0.000 description 1
- 210000001218 blood-brain barrier Anatomy 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 210000003792 cranial nerve Anatomy 0.000 description 1
- 229960003914 desipramine Drugs 0.000 description 1
- 229960000632 dexamfetamine Drugs 0.000 description 1
- MHUWZNTUIIFHAS-CLFAGFIQSA-N dioleoyl phosphatidic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(COP(O)(O)=O)OC(=O)CCCCCCC\C=C/CCCCCCCC MHUWZNTUIIFHAS-CLFAGFIQSA-N 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 208000024732 dysthymic disease Diseases 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 235000003969 glutathione Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000007952 growth promoter Substances 0.000 description 1
- 230000000971 hippocampal effect Effects 0.000 description 1
- 210000001320 hippocampus Anatomy 0.000 description 1
- 230000002631 hypothermal effect Effects 0.000 description 1
- BCGWQEUPMDMJNV-UHFFFAOYSA-N imipramine Chemical compound C1CC2=CC=CC=C2N(CCCN(C)C)C2=CC=CC=C21 BCGWQEUPMDMJNV-UHFFFAOYSA-N 0.000 description 1
- 229960004801 imipramine Drugs 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 229960004502 levodopa Drugs 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229960001344 methylphenidate Drugs 0.000 description 1
- 230000005709 nerve cell growth Effects 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000007472 neurodevelopment Effects 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The present invention relates to technical field of pharmaceuticals, particularly relate to a kind of pharmaceutical composition and application thereof.The pharmaceutical composition that the present invention provides includes: tomoxetine hydrochloride and taurine.The two combination can produce synergistic function.The present invention is found through experiments; contrast individually gives tomoxetine hydrochloride; taurine is used to be combined with tomoxetine hydrochloride; can more dramatically increase SOD activity and GSH content in juvenile stage ADHD rat model cerebral tissue, lower MDA content (P < 0.01); serve antioxidative effect; improve DBH and TH in juvenile stage ADHD rat model brain to express, raise Fos protein expression (P < 0.01) in Prefrontal Cortex cortex.Compared with individually dosed, the effect of pharmaceutical composition becomes apparent from.Therefore, the pharmaceutical composition of the present invention can be as the drug candidate for the treatment of hyperkinetic syndrome.
Description
Technical field
The present invention relates to technical field of pharmaceuticals, particularly relate to a kind of pharmaceutical composition and application thereof.
Background technology
Hyperkinetic syndrome, also known as hyperkinetic syndrome (ADHD) or minimal brain dysfunciton syndrome, is a kind of common
Child be abnormal diseases, main impact is to cause children's's dystropy.The main clinical manifestation of hyperkinetic syndrome is: note barrier
Hinder, hyperkinesia, perceptual disturbance, dysthymic disorder, maladjustment, learning difficulty etc..Its pathogenic factor probably has following several: brain
Neurodevelopment is bad, brain neurotransmitter lazy weight, cerebral tissue organic lesion, inherited genetic factors, other factors etc..It is mainly in
Virgin school age, the age bracket that prevalence is the highest be 6-12 year.
The most a lot of researchs show that children's attention defect hyperkinetic syndrome (ADHD) is mainly due to DOPA in central nervous system
Slight cerebral function defect caused by amine (DA) and norepinephrine (NA) dysbolismus.Research in recent years shows: oxidative stress is
One of child's ADHA pathogenesis, and SOD (superoxide dismutase), GSH (glutathion), MDA (malonaldehyde), BDNF (brain
Derived neurotrophic factor), AChE (acetylcholinesterase), DBH (dopamine-β-hydroxylase), TH (TYR hydroxylase),
Fos (oligofructose) is the biochemical indicator relevant to oxidative stress.Improve DBH and TH and Fos protein expression on nor-kidney
Parathyrine serotonergic neuron and dopaminergic neuron have protective effect.
And the medicine of hyperkinetic syndrome substantially can be divided into non-central stimulant, central nervous excitation agent, resist melancholy agent,
A few class such as psychosis and Anti-epileptics, concrete clinical application has tomoxetine hydrochloride, methylphenidate, dextro-amphetamine, benzene
Different appropriate English, caffeine, imipramine etc..
Tomoxetine hydrochloride (trade name is selected think of and reached), is strong presynaptic norepinephrine transporter inhibitor,
The energy Selective depression presynaptic amine pump reuptake to norepinephrine, thus improve the symptom of ADHD, indirectly promote understanding
Complete and the concentration of attention.But tomoxetine hydrochloride is the most weak to other monoamine transporters or receptor affinity, it is impossible to solve
Certainly hyperkinetic syndrome children's's brain neuroblastoma grows problem, the problem that there is " curing the symptoms, not the disease ".
Taurine (Taurine), also known as 2-aminoethyl sulfonic acid, is the non-protein of a kind of sulfur-bearing, deposits with free state in vivo
, it is not involved in the biosynthesis of vivo protein.Taurine rich content in brain, widely distributed, nerveous system can be obviously promoted
The growth promoter of system and cell proliferation, differentiation, and in dose dependent, play an important role in cranial nerve cell growth course.
Finding in the taurine animal experiment study with brain development relation, taurine can promote the learning and m emaory of rat.
Supplement appropriate taurine to be possible not only to improve learning and memory speed, but also the accuracy of learning and memory can be improved, and right
Neural defying age also has certain effect.
But, it is not used for treating grinding of hyperkinetic syndrome about by tomoxetine hydrochloride and taurine
Study carefully, and study the two combination, for treating, hyperkinetic syndrome is had good prospect and using value.
Summary of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of pharmaceutical composition and application, this medicine
Compositions includes tomoxetine hydrochloride and taurine, and the two combination can produce synergistic function.By increasing in cerebral tissue
SOD activity and GSH content, lower MDA content, raises DBH, TH or Fos and expresses, thus reaches to treat the mesh of hyperkinetic syndrome
's.
The invention provides a kind of pharmaceutical composition, including: tomoxetine hydrochloride and taurine.
As preferably, in the pharmaceutical composition that the present invention provides, tomoxetine hydrochloride is 1:0.5 with the mass ratio of taurine
～50.
Preferably, in the pharmaceutical composition that the present invention provides, tomoxetine hydrochloride is 1:5～45 with the mass ratio of taurine.
The pharmaceutical composition that the present invention provides is in the application prepared in brain in SOD accelerative activator.
The pharmaceutical composition that the present invention provides is in the application prepared in brain in MDA inhibitor.
The pharmaceutical composition that the present invention provides is the application in the medicine of GSH content in preparation increases brain.
The present invention is experimentally confirmed, and individually gives tomoxetine hydrochloride, can increase juvenile stage ADHD rat model brain
SOD activity and GSH content, downward MDA content (P < 0.05) in tissue, and use taurine and tomoxetine hydrochloride to be combined, can
More dramatically increase SOD activity and GSH content in juvenile stage ADHD rat model cerebral tissue, lower MDA content (P < 0.01), rise
Arrive antioxidative effect,
The pharmaceutical composition that the present invention provides application in the medicine that preparation promotes DBH, TH or Fos to express.
The present invention is experimentally confirmed, and individually gives tomoxetine hydrochloride, can improve juvenile stage ADHD rat model big
In brain, DBH and TH expresses, and raises Fos protein expression (P < 0.05) in Prefrontal Cortex cortex, and uses taurine and hydrochloric acid torr
Moses spit of fland is combined, and can more significantly improve DBH and TH in juvenile stage ADHD rat model brain and express, raise Prefrontal Cortex cortex
Middle Fos protein expression (P < 0.01).Show taurine and the tomoxetine combination norepinephrine energy to ADHD rat model
Neuron and dopaminergic neuron have protective effect.
The pharmaceutical composition that the present invention provides application in the medicine of preparation treatment hyperkinetic syndrome.
Owing to SOD activity and GSH content, MDA level, BDH, TH or Fos express closely related with hyperkinetic syndrome, at this
The pharmaceutical composition that invention provides can promote SOD activity, raising GSH content, suppression MDA level, promote BDH, TH or Fos table
Reaching, therefore, the pharmaceutical composition that the present invention provides can be used in the medicine of preparation treatment hyperkinetic syndrome.And the present invention is by real
Checking is real, and compared with individually giving tomoxetine hydrochloride, taurine can significantly (P < 0.05) reduce with tomoxetine hydrochloride combination
The rat spontaneous hyperkinetic behavior in new environment.
A kind of medicine treating hyperkinetic syndrome, the pharmaceutical composition provided including the present invention and medicine are provided
Acceptable adjuvant on.
As preferably, the medicine of the treatment hyperkinetic syndrome that the present invention provides is oral formulations.
Preferably, oral formulations is tablet, capsule, pill, granule, decoction, unguentum, distillate medicinal water, oral solutions, drop pill
Agent or syrup.
It is furthermore preferred that oral formulations is tablet.
As preferably, pharmaceutically acceptable adjuvant is excipient, filler and lubricant.
Preferably, excipient is mannitol；Filler is starch；Lubricant is magnesium stearate and micropowder silica gel.
As preferably, in the medicine of the treatment hyperkinetic syndrome that the present invention provides, the mass fraction of tomoxetine hydrochloride is
2%～50%；The mass fraction of described taurine is 2%～50%.
Preferably, the mass fraction of tomoxetine hydrochloride is 16.5%～18.9%；The mass fraction of described taurine is
14.2%～16.5%.
As preferably, in the medicine of the treatment hyperkinetic syndrome that the present invention provides, the mass parts of the most each component is:
Tomoxetine hydrochloride 100 parts；
Taurine 75 parts～100 parts；
275 parts～300 parts of mannitol；
Starch 75 parts～100 parts；
Magnesium stearate 2.5 parts～3 parts；
Micropowder silica gel 1.5 parts～2 parts.
As preferably, the dosage of the medicine of the treatment hyperkinetic syndrome that the present invention provides is 10mg/kg every day.
The invention provides a kind of pharmaceutical composition, including: tomoxetine hydrochloride and taurine.The two combination can produce association
Same potentiation.The present invention is experimentally confirmed, and compared with individually giving tomoxetine hydrochloride, uses taurine and atomoxetine hydrochloride
Xi Ting is combined, and significantly (P < 0.01) can increase SOD activity and GSH content energy in juvenile stage ADHD rat model cerebral tissue
Enough notable (P < 0.01) lowers MDA level, serves antioxidative effect, and significantly (P < 0.01) can improve juvenile stage ADHD mould
In type rat brain, DBH and TH expresses, and notable (P < 0.01) raises Fos protein expression in Prefrontal Cortex cortex.Show cattle
Sulfonic acid has guarantor with tomoxetine combination to noradrenergic neuron and the dopaminergic neuron of ADHD rat model
Protect effect.Therefore, present invention also offers this pharmaceutical composition for preparing the medicine for the treatment of hyperkinetic syndrome.
Accompanying drawing explanation
Fig. 1 shows the DBH SABC testing result of different disposal SHR rat ADHD model；Wherein: Fig. 1-a shows negative right
According to group rat DBH SABC testing result；Fig. 1-b shows blank group rat DBH SABC testing result；Fig. 1-c shows
Positive controls 1 rat DBH SABC testing result；Fig. 1-d shows positive controls 2 rat DBH SABC testing result；
Fig. 1-e shows experimental group 1 rat DBH SABC testing result；
Fig. 2 shows the Fos SABC testing result of different disposal SHR rat ADHD model；Wherein: Fig. 2-a shows negative right
According to group rat Fos SABC testing result；Fig. 2-b shows blank group rat Fos SABC testing result；Fig. 2-c shows
Positive controls 1 rat Fos SABC testing result；Fig. 2-d shows positive controls 2 rat Fos SABC testing result；
Fig. 2-e shows experimental group 1 rat Fos SABC testing result；
Fig. 3 shows different disposal 6-OHDA neonate rat ADHD model TH SABC testing result；Wherein: Fig. 3-a shows the moon
Property control rats TH SABC testing result；Fig. 3-b shows blank group rat TH SABC testing result；Fig. 3-c
Show positive controls 1 rat TH SABC testing result；Fig. 3-d shows positive controls 2 rat TH SABC testing result；
Fig. 3-e shows experimental group 1 rat TH SABC testing result；
Fig. 4 shows the Fos SABC testing result of different disposal 6-OHDA neonate rat ADHD model: wherein: Fig. 4-a shows
Negative control group rat Fos SABC testing result；Fig. 4-b shows blank group rat Fos SABC testing result；Figure
4-c shows positive controls 1 rat Fos SABC testing result；Fig. 4-d shows that positive controls 2 rat Fos SABC detects
Result；Fig. 4-e shows experimental group 1 rat Fos SABC testing result.
Detailed description of the invention
The invention provides a kind of pharmaceutical composition and application thereof, those skilled in the art can use for reference present disclosure, suitable
Realize when improving technological parameter.Special needs to be pointed out is, all similar replacements and change are for a person skilled in the art
Being apparent from, they are considered as being included in the present invention.Method and the application of the present invention are entered by preferred embodiment
Having gone description, methods herein and application substantially can be carried out in without departing from present invention, spirit and scope by related personnel
Change or suitably change and combination, realize and apply the technology of the present invention.
The instrument that the present invention uses is all common commercially available product, all can buy in market.
Below in conjunction with embodiment, the present invention it is expanded on further:
The pharmaceutical composition that embodiment 1 present invention provides impact on juvenile stage SHR rat ADHD model
1), experimental design
Take 54, newborn 21 days SHR rat ADHD models and WKY rat 9, after adaptability is fed 3 days, select 52 SHR
Rat is classified into 6 groups at random by body weight, 8～9/group.
Experiment is with WKY rat as negative control；
With the SHR rat ADHD model that is not administered as blank；
Be given only tomoxetine hydrochloride rat as positive controls；
And set three experimental grouies of different dosing dosage: wherein, experimental group 1 gives the pharmaceutical composition of high dose, experiment
Group 2 gives the pharmaceutical composition of middle dosage, experimental group 3 gives the pharmaceutical composition of low dosage.Dosage is as shown in table 1:
According to table 1 dosage, every day gastric infusion once, successive administration 2 weeks (PD23～PD37).Administration volume is all
10mL/kg, negative control group and blank group give equal-volume distilled water.Within every three days, weigh, adjust by body weight and be administered body
Long-pending.
The packet of table 1 animal and dosage regimen
It is grouped and dosage regimen according to table 1, carries out the open field test of juvenile stage SHR rat, Y maze experiment, and detect SHR
The physiology of rat, biochemical indicator, analyze the pharmaceutical composition providing the present invention impact on SHR rat ADHD model.
2), the pharmaceutical composition that the present invention the provides impact on juvenile stage SHR rat ADHD model open field test
Open-field system (Panlab company, Spain) is used to test the hyperkinetic behavior of each group of juvenile stage SHR rat.
The experiment of spacious field is carried out in the room of 30 DEG C of constant temperature, sound insulation and low-light level.Rat is put into the spacious field experimental box (120cm of uncovered
× 120cm × 50cm) center after start timing, photographic head records the motion path of rat automatically, and every animal is surveyed continuously
Try 5 minutes.By every animal in Smart3.0 computed in software 5 minutes enter middle section (60cm × 60cm) number of times and
Percentage ratio in the middle section time of staying.Result represents with mean ± standard deviation, and SPSS16.0 software one-way ANOVA divides
Significant difference between analysis group, result is as shown in table 2.
The pharmaceutical composition that table 2 present invention provides impact on juvenile stage SHR rat ADHD model open field test
| Group | Enter the number of times (secondary) of central area | In central area time of staying percentage ratio (%) |
| Negative control | 12.3±1.5** | 2.1±0.8** |
| Blank | 101.8±3.0 | 18.4±3.1 |
| Positive controls | 66.6±1.8* | 12.8±2.4* |
| Experimental group 1 | 23.2±1.5** | 5.1±1.2** |
| Experimental group 2 | 25.1±1.8** | 5.9±1.0** |
| Experimental group 3 | 26.3±1.1** | 7.4±1.5** |
Note: * shows that P < 0.05, * * shows P < 0.01 compared with blank group compared with blank group
As shown in table 2, compared with blank group, it is used alone tomoxetine hydrochloride, can notable (P < 0.05) reduce
Rat enters spacious field experimental box central area number of times and at central area time of staying percentage ratio.The pharmaceutical composition energy that the present invention provides
More notable (P < 0.01) reduces rat and enters spacious field experimental box central area number of times and at central area time of staying percentage ratio.Show this
The pharmaceutical composition that invention provides can significantly inhibit the hyperkinetic behavior of SHR rat ADHD model, the effect that two medicine combinations produce
More independent medication becomes apparent from, and illustrates that compositions can produce synergistic function.
3), the pharmaceutical composition that the present invention the provides impact on juvenile stage SHR rat ADHD model Y maze experiment
Use the ability of learning and memory of Y maze experiment fixed number of times random not rest method test rat.MG-2C/3C type Y fan
Palace (Biomedical Instruments factory of Zhangjagang City) is made up of controller and Y type case (I, II, III 3 isometric labyrinth arms) of getting lost.Case
The end, is laid by electricity grid and forms, and every arm top is equipped with stimulus signal lamp.Controller has the button of regulation output voltage and for selecting
Four buttons (I, II, III, 0) of labyrinth arm and junctional area.Press I, II at random, during III button, the stimulus signal lamp of respective arms
Bright, this arm is cold place of safety, and the junctional area being energized and other two-arm are non-security district.When pressing 0, junctional area leads to
Electricity, three arms all no powers.When training experiment starts, randomly choosing an arm is place of safety, rat is put into adaptation 2 minutes, subsequently
Press other two-arm button at random, can escape after rat irriate, rat is directly gone to place of safety and is defined as " correct response ",
Other situations are " wrong reaction " (such as: run to without light district or inversely run).Follow-on test 10 times.After 24 hours, with advancing
The formal experiment of row, measures rat correct response number of times, evaluates rat with accuracy and escapes the ability of learning and memory of electricity irritation:
Accuracy=correct response number of times/10 × 100%
Result represents with mean ± standard deviation, the significant difference between SPSS16.0 software one-way ANOVA analysis group,
Result is as shown in table 3:
The pharmaceutical composition that table 3 present invention provides impact on juvenile stage SHR rat ADHD model Y maze experiment
| Group | Accuracy (%) |
| Negative control | 93.5±2.2** |
| Blank | 36.7±1.9 |
| Positive controls | 52.3±2.5* |
| Experimental group 1 | 75.9±2.5** |
| Experimental group 2 | 76.5±3.1** |
| Experimental group 3 | 78.8±2.4** |
Note: * shows that P < 0.05, * * shows P < 0.01 compared with blank group compared with blank group
As shown in table 3, compared with blank group, it is used alone tomoxetine hydrochloride, it is possible to notable (P < 0.05) carries
The accuracy of high rat Y maze experiment, the pharmaceutical composition that the present invention provides, it is possible to more notable (P < 0.01) improves rat Y fan
The accuracy of palace experiment, the more independent medicine of effect of compositions becomes apparent from.Show that two medicine combinations produce obvious Synergistic and make
With.
4), the pharmaceutical composition that the present invention the provides impact on juvenile stage SHR rat ADHD model physiological index
After last is administered 24 hours, weigh.Use RBP-1B type rat blood pressure instrument (Sino-Japan close friend's clinic study
Institute) noinvasive tail sleeve method measure Conscious Rat blood pressure and heart rate.Rat is put in blood pressure instrument preheating 5 minutes, by blood-pressure measuring appliance
Pressure arteries and veins set set to rat tails proximal part, transducer be placed in tail at upper 1/3, regulate amplifier sensitivity and make pulse wave to 2-
3cm is advisable, and after regular pulse wave occurs, makes the interior pressure of pressure arteries and veins set be increased to pulse wave by inflatable ball and is wholly absent, then
Pressurization raises 30mmHg, is then slowly exitted by balloon valve, is 0 holding 5-6 second from starting to be deflated to manifold pressure.Young
Carefully observing and read pulse wave from without to force value corresponding when just starting to occur first ripple, this, for shrinking pressure, reads simultaneously
Heart rate value on monitor.Result represents with mean ± standard deviation, the system between SPSS16.0 software one-wayANOVA analysis group
Difference learned by meter, the results are shown in Table 4.
The pharmaceutical composition that table 4 present invention provides impact on juvenile stage SHR rat ADHD model physiological index
As shown in table 4, the physiology of juvenile stage SHR rat ADHD model is not referred to by the pharmaceutical composition that giving the present invention provides
Mark makes a significant impact.
5), the pharmaceutical composition that the present invention the provides impact on juvenile stage SHR rat ADHD model biochemical indicator
A) cerebral tissue specimen collection: take each group through open field test and the rat of Y maze experiment, deep with 3.5% chloral hydrate
Degree anesthesia, opens thoracic cavity, 4 DEG C of normal saline of cardiac perfusion, takes cerebral tissue, and left side cerebral tissue is frozen in-80 DEG C, for biochemistry
Indexs measure；Right side brain is fixed on formalin, and routine paraffin wax embedding, section, for Immunohistochemical detection.
B) biochemical indicator detection: oxidative stress is one of child's ADHA pathogenesis, and therefore, the present invention have detected and aoxidizes
The biochemical indicator that stress be correlated with.The test kit description that building up Bioengineering Research Institute according to Nanjing provides measures cerebral tissue
MDA Yu GSH assay and SOD Yu AChE enzymatic activity；Measure big according to Xi Tang bio tech ltd, Shanghai description
Cerebral tissue BDNF content, and calculate each index relative amount compared with blank group.Statistical result is as shown in table 5:
The pharmaceutical composition that table 5 present invention provides impact on juvenile stage SHR rat ADHD model biochemical indicator
Note: * shows that P < 0.05, * * shows P < 0.01 compared with blank group compared with blank group
As shown in table 5, compared with blank group, it is used alone tomoxetine hydrochloride, SOD activity can be dramatically increased,
GSH content, minimizing MDA content (P < 0.05), and the pharmaceutical composition that the present invention provides more notable (P < 0.01) improves rat brain
SOD activity in tissue, GSH content, reduce MDA content.Therefore illustrate, and be used alone tomoxetine hydrochloride or taurine phase
Ratio, the antioxidation that the pharmaceutical composition that the present invention provides produces becomes apparent from, and two medicine combinations produce synergism.
6), immunohistochemical experiment identifies that the pharmaceutical composition of present invention offer is to juvenile stage SHR rat ADHD modeling brain
The impact of BDH, TH and Fos in cortex
Each group of rat brain paraffin section is positioned in 55 DEG C of baking boxs roasting 2 hours, is placed in dimethylbenzene stock solution (I, II, III
Three parts) in dewax respectively 2 minutes, then with each 1 minute of gradient concentration ethanol from high to low (100%～70%) rehydration, distilled water
Clean 5 minutes × 3 times.H2O2 (fresh configuration) the at room temperature lucifuge of 3% is hatched 20 minutes, in order to inactivating endogenous peroxidating
Thing enzyme, distilled water cleans 5 minutes × 3 times.Put antigen retrieval buffers mesohigh to repair 3 minutes, place to natural cooling after taking-up.
PBS 5 minutes × 3 times, drips 5%BSA confining liquid, is placed in 37 DEG C of incubators in wet box and hatches 1 minute.Suck around tissue many
Remaining liquid, dropping PBS liquid dilutes an anti-DBH (1:100) or Fos (1:50) or TH (1:50), 37 DEG C hatch 1 hour after dislocation in
4 DEG C of refrigerator overnight.PBS liquid cleans 5 minutes × 3 times.Dropping biotinylation two resists, and hatches 1 hour in 37 DEG C.PBS 5 minutes
× 3 times.Blotting PBS, dropping dropping biotinylation goat anti-rabbit igg two resists, and hatches 1 hour for 37 DEG C.PBS 5 minutes × 3 times.
Dropping DAB developer, examines under a microscope and controls chromogenic reaction, terminates reaction with distilled water in good time.Haematoxylin carries out multiple
Dye；Hydrochloride alcohol breaks up, and tap water returns indigo plant；Conventional gradients ethanol (75%～100%) is dehydrated, dimethylbenzene is transparent, neutral gum
Mounting.Under microscope, blind observes ImmunohistochemistryResults Results, according to experimenter's early stage semi-quantitative method, in lower point of high power lens (× 40)
Do not count TH-positive neuronal cell number and 3, Prefrontal Cortex cortex brain district under 3 visuals field, each Brain striatal brain district to regard
Yezhong DBH-positive neuronal cell number and Fos-positive cell number (Yu, et al., 2013).Result is with mean ± standard deviation
Representing, the significant difference between SPSS16.0 software one-way ANOVA analysis group, result is shown in Fig. 1～2.Statistical result such as table 6
Shown in:
The pharmaceutical composition that table 6 present invention provides is to BDH, TH and Fos in juvenile stage SHR rat ADHD modeling brain cortex
Impact
| Group | DBH positive cell number | TH positive cell number | Fos positive cell number |
| Negative control | 52.3±1.0** | 17.9±0.8** | 28.6±1.2** |
| Blank | 16.7±1.5 | 2.7±0.6 | 7.5±0.9 |
| Positive controls | 31.2±1.8* | 6.6±1.0* | 17.6±1.1* |
| Experimental group 1 | 45.1±1.4** | 12.8±0.5** | 27.3±1.1** |
| Experimental group 2 | 43.2±1.1** | 12.5±0.6** | 25.5±1.3** |
| Experimental group 3 | 48.3±1.0** | 13.5±0.8** | 26.3±0.9** |
Note: * shows that P < 0.05, * * shows P < 0.01 compared with blank group compared with blank group
As shown in table 5, compared with blank group, being used alone tomoxetine hydrochloride can significantly (P < 0.05) raise
Brain DBH, TH and Fos express, and the pharmaceutical composition that the present invention provides more notable (P < 0.01) can raise brain DBH, TH
Express with Fos, prompting tomoxetine hydrochloride, taurine or taurine and the tomoxetine combination noradrenaline to ADHD rat
Element serotonergic neuron and dopaminergic neuron have protective effect, and two medicine combinations are closed medicine and compared with independent medicine, its effect
More notable.The pharmaceutical composition that embodiment 2 present invention provides affects 1 to 6-OHDA neonate rat ADHD model), animal model
And packet is administered
When neonate rat 5 days, be randomly divided into 7 groups, be followed successively by: Normal group, model control group, positive controls,
Experimental group 1～3.
In addition to Normal group, remaining respectively organizes subcutaneous injection 0.1ml/10g hydrochloric acid desipramine solution (2.5mg/ respectively
Ml), after 40 minutes, modeling rat is in hypothermic anesthesia on ice 5 minutes, l/ 6-OHDA solution (5mg/ml) of intracisternal injection 20 μ；
Rats in normal control group is after anaesthetizing 5 minutes on ice, and equal-volume solvent is injected in brain pond.
Each group administrations is as shown in table 7, every day gastric infusion once, successive administration 21 days (PD5～PD26).It is administered body
Amass and be all 0.05mL/10g.Within every three days, weigh, adjust by body weight and be administered volume.
The packet of table 7 animal and dosage regimen
2), the pharmaceutical composition that the present invention the provides impact on 6-OHDA neonate rat ADHD model body weight
After being administered 21 days, weigh.Result represents with mean ± standard deviation, and SPSS16.0 software one-way ANOVA divides
Significant difference between analysis group, the results are shown in Table 8.
The pharmaceutical composition that table 8 present invention provides impact on 6-OHDA neonate rat ADHD model body weight
Note: * * shows and P < 0.01 compared with Normal group
As shown in table 8, after being administered 21 days, the body weight of 6-OHDA model group rats substantially alleviates (P < 0.01)；PD26, single
Give tomoxetine (1mg/kg) and body weight is had no impact；PD5～PD26 give respectively 33mg/kg/d, 100mg/kg/d or
The 6-OHDA-rat of 300mg/kg/d taurine, body weight all has > growth of 10%.
3), the pharmaceutical composition that the present invention the provides impact on 6-OHDA neonate rat ADHD model hyperkinetic behavior
After tomoxetine is administered 1 hour, open field test is used to evaluate the rat spontaneous hyperkinetic behavior in new environment movable
Property.Prologue proof box (100cm × 100cm × 50cm) is uncovered black experimental box, by white line divide 25 grids (20cm ×
20cm) composition.When experiment starts, rat being put into case, record rat wore lattice number of times in 5 minutes.Result is with mean ± standard
Difference represents, the significant difference between SPSS16.0 software one-way ANOVA analysis group.Result is as shown in table 9.
The pharmaceutical composition that table 9 present invention provides impact on 6-OHDA neonate rat ADHD model hyperkinetic behavior
| Group | Wear lattice number of times (secondary) |
| Normal control | 48.5±1.3 |
| Model comparison | 100.2±1.9## |
| Positive controls | 55.6±1.5* |
| Experimental group 1 | 33.1±1.1** |
| Experimental group 2 | 34.3±1.0** |
| Experimental group 3 | 31.8±1.3** |
##Show P < 0.01 compared with Normal group,*Show P < 0.05 compared with model group,**Show compared with model group
P<0.01
As shown in table 9, compared with Normal group, model group rats is worn lattice number of times and is dramatically increased (P < 0.01).With mould
Type matched group compares, and being used alone tomoxetine hydrochloride significantly (P < 0.05) can reduce rat and wear lattice number of times, and the present invention
The pharmaceutical composition provided more notable (P < 0.01) reduces rat and wears lattice number of times.Show that the pharmaceutical composition that the present invention provides can
Significantly inhibit the hyperkinetic behavior of SHR rat ADHD model.
4), the present invention provides the pharmaceutical composition impact on 6-OHDA neonate rat ADHD model biochemical indicator
A) cerebral tissue specimen collection: take each group wear long lattice experiment rat, 3.5% chloral hydrate deep anaesthesia, open thoracic cavity,
4 DEG C of normal saline of cardiac perfusion, take cerebral tissue, and left side cerebral tissue is frozen in-80 DEG C, detect for biochemical indicator；Right side
Brain is fixed on formalin, and routine paraffin wax embedding, section, for Immunohistochemical detection.
B) biochemical indicator detection: the test kit description that building up Bioengineering Research Institute according to Nanjing provides measures brain group
Knit MDA content；Cerebral tissue BDNF content, result such as table 10 institute is measured according to Xi Tang bio tech ltd, Shanghai description
Show.
The pharmaceutical composition that table 10 present invention provides impact on 6-OHDA neonate rat ADHD model biochemical indicator
##Show P < 0.01 compared with Normal group,*Show P < 0.05 compared with model group,**Show compared with model group
P<0.01
As shown in table 10, compared with model group, it is used alone tomoxetine hydrochloride and can reduce MDA content (P > 0.05),
Raise BDNF content (P < 0.05).And the pharmaceutical composition that the present invention provides significantly reduces MDA content in rat cerebral tissue, raise
BDNF content.Result shows, compared with being used alone tomoxetine hydrochloride, the pharmaceutical composition that the present invention provides can produce more
Significantly antioxidation, two medicines share and compare with independent medicine, and its effect is more notable.
5), immunohistochemical experiment identifies that the pharmaceutical composition of present invention offer is to 6-OHDA neonate rat ADHD
The impact of BDH, TH and Fos in modeling brain cortex
The brain paraffin section of each group of rat is positioned in 55 DEG C of baking boxs roasting 2 hours, be placed in dimethylbenzene stock solution (I, II,
III 3 parts) in dewax respectively 2 minutes, then with each 1 minute of gradient concentration ethanol from high to low (100%～70%) rehydration, double steamings
Water cleans 5 minutes × 3 times.The H of 3%2O2(fresh configuration) at room temperature lucifuge is hatched 20 minutes, in order to inactivating endogenous peroxide
Compound enzyme, distilled water cleans 5 minutes × 3 times.Put antigen retrieval buffers mesohigh to repair 3 minutes, place to natural cooling after taking-up.
PBS 5 minutes × 3 times, drips 5%BSA confining liquid, is placed in 37 DEG C of incubators in wet box and hatches 1 minute.Suck around tissue many
Remaining liquid, dropping PBS liquid dilutes an anti-Fos (1:50) or TH (1:50) or an anti-DBH (1:100), 37 DEG C hatch 1 hour after move
It is placed in 4 DEG C of refrigerator overnight.PBS liquid cleans 5 minutes × 3 times.Dropping biotinylation two resists, and hatches 1 hour in 37 DEG C.PBS 5
Minute × 3 times.Blot PBS, drip SABC, hatch 1 hour for 37 DEG C.PBS 5 minutes × 3 times.Dropping DAB developer, aobvious
Micro-Microscopic observation also controls chromogenic reaction, terminates reaction with distilled water in good time.Haematoxylin is redyed；Hydrochloride alcohol breaks up, from
Water returns indigo plant；Conventional gradients ethanol (75%～100%) is dehydrated, dimethylbenzene is transparent, neutral gum mounting.Under microscope, blind is seen
Examine ImmunohistochemistryResults Results, under high power lens (× 40), count TH-positive neurons under 3 visuals field, each Brain striatal brain district respectively
Fos-positive cell number and DBH-positive neuronal cell number (Yu, et under unit's cell number and the visual field, 3, prefrontal cortex brain district
al.,2013).Result represents with mean ± standard deviation, and the statistics between SPSS16.0 software one-way ANOVA analysis group is poor
Different, result is shown in Fig. 3～4, and statistical result is as shown in table 11:
The pharmaceutical composition that table 11 present invention provides to BDH, TH in 6-OHDA neonate rat ADHD modeling brain cortex and
The impact of Fos
* show that P < 0.05, * * shows P < 0.01 compared with blank group compared with blank group
As shown in table 11, compared with blank group, negative control group BDH, TH, Fos express dramatically increase (P <
0.01), and being used alone tomoxetine hydrochloride, the expression to appeal three has no significant effect, and the medicine group that the present invention provides
Compound significantly (P < 0.01) can raise brain DBH, TH and Fos expression, prompting taurine or taurine and tomoxetine combination
Noradrenergic neuron and dopaminergic neuron to ADHD rat have protective effect, and two medicine combinations produce substantially
Synergistic function.
Pharmaceutical composition pharmacokinetics and brain distribution that embodiment 3 present invention provides are studied
Inside the Pass " drug registration management method " and " chemical drugs Non-clinical Pharmacokinetics research guideline " phase
Hold requirement, with SD rat as study subject, alone to tomoxetine hydrochloride and combination taurine after pharmacokinetics and brain distribution at the beginning of
Step research.
Take SD rat 10 and be only randomly divided into A, B bis-groups, fasting 12 hours before experiment.A group single oral gives atomoxetine hydrochloride
Xi Ting, dosage is 5mg/kg；B group single oral is combined and is given tomoxetine hydrochloride and taurine, dosage be respectively 5mg/kg,
0.9g/kg.In administration process, ensure that SD rat is taken medicine completely.Gather blank blood before being administered, after administration in 0.033,0.083,
0.25,0.5,1,2,3,5,7,9,12 and 24h tail vein blood 0.3ml, blood sample is put in heparinization blood taking tube, and 8000rpm is centrifuged
5min, separated plasma, to be measured in-40 DEG C of Refrigerator stores.
Separately take SHR rat model 18 and be only randomly divided into the big group of G, H two, fasting 12 hours before experiment.G group single oral gives
Tomoxetine hydrochloride, dosage is 5mg/kg；H group single oral is combined and is given tomoxetine hydrochloride and taurine, and dosage is respectively
5mg/kg and 0.9g/kg.0.5,3 and 7h sacrificed by decapitation rats after being administered, three rats of each time point.Divided in 1 minute
Taking cerebral tissue, with normal saline drip washing surface, filter paper blots, and weighs, and puts in homogenizer, by cerebral tissue: normal saline=1:1
(m/m) normal saline is added, homogenate.It is to be measured that homogenate puts-40 DEG C of Refrigerator stores.
LC/MS/MS method is used to measure the tomoxetine hydrochloride in each sample and content of taurine, plasma sample room temperature solution
Freezing, accurate 100 μ l blood plasma of drawing, in 1.5ml centrifuge tube, add inner mark solution (768ng mL-1) 10 μ l, flowing 10 μ l mutually,
Formic acid 20 μ l, vortex 1min mix.Adding ethyl acetate: dichloromethane (4:1) 1ml, vortex 3min mix, 1200rpm is centrifuged
5min, in Aspirate supernatant to tool plug centrifuge tube, Nitrogen evaporator 37.5 DEG C dries up, and residue redissolves mutually with 100 μ l flowings, vortex
1min, 1200rpm are centrifuged 5min, take supernatant 20 μ l sample introduction.
Brain tissue sample's same treatment.
Detection method is:
Chromatographic condition
Flowing phase: methanol (0.2% formic acid): water (containing 0.02mol/l ammonium acetate) (80:20, V/V)；
Chromatographic column: Inertsil ODS-3 (100 × 2.1mm, 5 μm)；
Flow velocity: 300 μ l/min；
Column temperature: 30 DEG C；
Sample size: 20 μ l；
Mass Spectrometry Conditions
Ion detection mode (Scan Type): MRM；
Ion polarity (Polarity): positive；
Ionization mode: ESI；
Medicine distribution testing result in cerebral tissue and blood plasma is as shown in table 12:
Distribution in table 12 tomoxetine hydrochloride cerebral tissue and plasma sample
Table 12 result shows: SD rat single oral tomoxetine hydrochloride gives tomoxetine hydrochloride and taurine with combining
After, measure tomoxetine hydrochloride concentration in cerebral tissue by LC/MS/MS method.Atomoxetine hydrochloride in Figure 12 is obtained after being arranged by the data obtained
Xi Ting scattergram in SD rat brain.From chart, after drug combination, at 0.25h, when 0.5h, 2h, 3h, 5h, SD rat
The concentration of tomoxetine hydrochloride during the concentration of tomoxetine hydrochloride is all higher than individually dosed rear SD rat cerebral tissue in cerebral tissue.Should
During result explanation drug combination, taurine may promote that tomoxetine hydrochloride passes through blood brain barrier and enters cerebral tissue.
Medicine dynamic experiment result shows: single oral tomoxetine hydrochloride gives tomoxetine hydrochloride and cattle sulphur with combining
After acid, the AUC0-T of tomoxetine hydrochloride is respectively 129.17 ± 39.637 μ g h L-1 and 128.96 ± 46.422 μ g
H L-1AUC0-∞ is respectively 140.608 ± 41.383 μ g h L-1 and 142.661 ± 44.445 μ g h L-1, Cmax
Being respectively 65.537 ± 34.847ng mL-1 and 128.96 ± 46.422ng mL-1, Tmax is respectively 0.307 ± 0.197 He
0.363 ± 0.405h, T1/2 are respectively 2.942 ± 0.706h and 3.378 ± 1.666h.
Through statistical analysis, AUC0-T, AUC0-∞, T of tomoxetine hydrochloride during individually dosed and drug combinationmaxWithout system
Meter learns difference (P > 0.05), CmaxThere are significant difference (P < 0.05), the C of tomoxetine hydrochloride after administering drug combinationsmaxIt is significantly less than
C after individually dosedmax.Result show drug combination compared with individually dosed, tomoxetine hydrochloride absorption in SD rat body
Degree is without significant difference.
The preparation of the medicine of the treatment hyperkinetic syndrome that embodiment 4 present invention provides
Weigh tomoxetine hydrochloride 100 grams, taurine 100 grams, 300 grams of mannitol, starch 100 grams, magnesium stearate 3 grams,
Micropowder silica gel 2 grams, is sufficiently mixed, and adds the stirring of appropriate distilled water and pelletizes, crosses 20 mesh sieves, be dried, tabletting, every agreement that contracts a film or TV play to an actor or actress 0.3g.
The preparation of the medicine of the treatment hyperkinetic syndrome that embodiment 5 present invention provides
Weigh tomoxetine hydrochloride 200 grams, taurine 150 grams, 550 grams of mannitol, starch 150 grams, magnesium stearate 5 grams,
Micropowder silica gel 3 grams, is sufficiently mixed, and adds the stirring of appropriate distilled water and pelletizes, crosses 20 mesh sieves, be dried, tabletting, every agreement that contracts a film or TV play to an actor or actress 0.3g.
The preparation of the medicine of the treatment hyperkinetic syndrome that embodiment 6 present invention provides
Weigh tomoxetine hydrochloride 200 grams, taurine 100 grams, 325 grams of mannitol, starch 125 grams, magnesium stearate 5 grams,
Micropowder silica gel 3 grams, is sufficiently mixed, and adds the stirring of appropriate distilled water and pelletizes, crosses 20 mesh sieves, be dried, pack, every bag of about 0.3g.
The preparation of the medicine of the treatment hyperkinetic syndrome that embodiment 7 present invention provides
Weigh tomoxetine hydrochloride 4 grams, taurine 200 grams, 315 grams of mannitol, starch 130 grams, magnesium stearate 4 grams, micro-
2.5 grams of powder silica gel, is sufficiently mixed, and adds the stirring of appropriate distilled water and pelletizes, crosses 20 mesh sieves, be dried, fill capsule about 0.3g.
The medicine effect detection of the treatment hyperkinetic syndrome that embodiment 8 present invention provides
Laboratory animal: selecting healthy male SHR rat 18, week old is 10 weeks, and body weight 265～295g, purchased from Sichuan
Province Academy of Medical Sciences institute of lab animals.
Packet is with administration: 18 male SHR rats are randomly divided into 3 groups, often group 6, and gastric infusion is respectively experimental group,
Positive controls and blank group, wherein, experimental group gives the medicine that embodiment 2 provides, and positive controls gives hydrochloric acid torr
Moses spit of fland, blank group gives isopyknic normal saline.Dosage is all 10mg/kg, and every morning gavages, 1 times/day,
Gavage 14 days continuously.During experiment, every day gives every rat 15～20g feedstuff, and water can freely obtain.
Detection method: unified after continuous gavage 14 days draw materials for above-mentioned each group, first numb with 10% chloral hydrate lumbar injection
Liquor-saturated rat, then by rat sacrificed by decapitation, takes rapidly brain on ice platform and isolates Hippocampus and cortex of frontal lobe tissue, being distinguished
Wrap with tinfoil after weighing, be stored in-70 DEG C of refrigerators, then high-efficient liquid phase technique detection norepinephrine (NE), 5-hydroxy tryptamine
(5-HT) content.
Statistical method: use SPSS11.5 software kit to carry out data compilation and statistical analysis.The neat person of measurement data variance
Check with t, heterogeneity of variance person's rank test, compare between group and use one factor analysis of variance.Result is as shown in table 13:
The medicine effect detection of the treatment hyperkinetic syndrome that table 13 present invention provides
| Group | Methylepinephrine (NE) | 5-hydroxy tryptamine (5-HT) |
| Experimental group | 23.65±8.69** | 12.24±5.63** |
| Positive controls | 45.24±3.21** | 9.24±13.92* |
| Blank group | 34.84±8.63 | 6.47±12.65 |
* show that P < 0.05, * * shows P < 0.01 compared with blank group compared with blank group
Result shows: compared with blank group, embodiment 2 provide tablet can significantly (P < 0.01) reduction SHR big
The content of NE in hippocampal tissue in Mus brain, raises the content of 5-HT in cortex of frontal lobe tissue, and positive control medicine can significantly raise
The content of the two, wherein NE (P < 0.01), 5-HT (P < 0.05).
Below it is only the preferred embodiment of the present invention, it is noted that those skilled in the art are come
Saying, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (10)
1. a pharmaceutical composition, it is characterised in that be made up of tomoxetine hydrochloride and taurine.
Pharmaceutical composition the most according to claim 1, it is characterised in that described tomoxetine hydrochloride and the quality of taurine
Ratio is 1:0.5～50.
3. pharmaceutical composition as claimed in claim 1 is in the application prepared in brain in SOD accelerative activator.
4. pharmaceutical composition as claimed in claim 1 is in the application prepared in brain in MDA inhibitor.
5. pharmaceutical composition as claimed in claim 1 application in the medicine of GSH content in preparation increases brain.
6. the application in the pharmaceutical composition medicine that DBH, TH or Fos express in preparation promotes brain as claimed in claim 1.
7. the pharmaceutical composition as claimed in claim 1 application in the medicine of preparation treatment hyperkinetic syndrome.
8. the medicine treating hyperkinetic syndrome, it is characterised in that include the medicine as described in any one of claim 1～2
Compositions and pharmaceutically acceptable adjuvant.
Medicine the most according to claim 8, it is characterised in that described pharmaceutically acceptable adjuvant is excipient, filling
Agent and lubricant.
Medicine the most according to claim 8, it is characterised in that the mass fraction of wherein said tomoxetine hydrochloride is 2%
～50%；The mass fraction of described taurine is 2%～50%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410267131.6A CN104055759B (en) | 2014-06-16 | 2014-06-16 | A kind of pharmaceutical composition and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410267131.6A CN104055759B (en) | 2014-06-16 | 2014-06-16 | A kind of pharmaceutical composition and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104055759A CN104055759A (en) | 2014-09-24 |
| CN104055759B true CN104055759B (en) | 2016-08-31 |
Family
ID=51543881
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410267131.6A Expired - Fee Related CN104055759B (en) | 2014-06-16 | 2014-06-16 | A kind of pharmaceutical composition and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104055759B (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001026642A2 (en) * | 1999-10-08 | 2001-04-19 | Joyce Corinne Bechthold | Methods and compositions for treating neurobehavioral disorders |
| US20050209303A1 (en) * | 2004-03-22 | 2005-09-22 | Gottfried Kellermann | Method of treatment for high neurotransmitter pattern |
| CA2840363A1 (en) * | 2011-06-24 | 2012-12-27 | K-Pax Pharmaceuticals, Inc. | Compositions comprising a central nervous system stimulant and select micronutrients useful in the treatment of chronic fatigue |
-
2014
- 2014-06-16 CN CN201410267131.6A patent/CN104055759B/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| 治疗ADHD的新药-托莫西汀;杜亚松;《中国儿童保健杂志》;20080229;第16卷(第1期);第65-66和69页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104055759A (en) | 2014-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sullivan et al. | Norepinephrine-induced plasticity and one-trial olfactory learning in neonatal rats | |
| Gatto et al. | Ethanol self-infusion into the ventral tegmental area by alcohol-preferring rats | |
| Segawa | Hereditary progressive dystonia with marked diurnal fluctuation | |
| Liu et al. | Environment-contact administration of rotenone: A new rodent model of Parkinson’s disease | |
| ES2573733T3 (en) | Pharmacological treatment of cognitive impairment | |
| Broadbent et al. | Rats depend on habit memory for discrimination learning and retention | |
| US8461389B2 (en) | Psycho-pharmaceuticals | |
| Ji et al. | Sigma-1 receptor activation ameliorates anxiety-like behavior through NR2A-CREB-BDNF signaling pathway in a rat model submitted to single-prolonged stress | |
| Yamamoto et al. | Differential activity of drugs to induce emesis and pica behavior in Suncus murinus (house musk shrew) and rats | |
| JP2018516906A (en) | Sustained release pharmaceutical composition of levetiracetam | |
| Chen et al. | Efficacy of apraclonidine 0.5% in the diagnosis of Horner syndrome in pediatric patients under low or high illumination | |
| Yamazaki et al. | FK960 N-(4-acetyl-1-piperazinyl)-p-fluorobenzamide monohydrate ameliorates the memory deficits in rats through a novel mechanism of action. | |
| WO2015073833A1 (en) | Methods for delaying or preventing the onset of type 1 diabetes | |
| CN109498643A (en) | A kind of folate composition and its application in the old mistake intelligence drug of preparation improvement | |
| US20210161886A1 (en) | Methods of treating depressive disorders | |
| CN104055759B (en) | A kind of pharmaceutical composition and application thereof | |
| Pautassi et al. | Acute ethanol counteracts the acquisition of aversive olfactory learning in infant rats | |
| BRPI0706865A2 (en) | Method for preparing shinyleaf yellowhorn extract and shinyleaf yellowhorn extract | |
| CN109718236B (en) | Application of bulleyaconitine A in preparing medicine for treating irritable bowel syndrome | |
| Patterson et al. | Toxicity assessment of pramipexole in juvenile rhesus monkeys | |
| Vasilev et al. | Age-related differences in MK-801-and amphetamine-induced locomotor and stereotypic activities of rats | |
| CN101627984A (en) | Application of 2-(alpha- hydroxyl amyl) potassium benzoate in preventing and/or treating senile dementia | |
| Pappas et al. | Neonatal brain dopamine depletion and the cortical and behavioral consequences of enriched postweaning environment | |
| CN105078952A (en) | Levodopa preparation and application thereof | |
| ES2747993T3 (en) | Levetiracetam prolonged-release pharmaceutical compositions |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160831 |