CN118236369A - Application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitor - Google Patents
Application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitor Download PDFInfo
- Publication number
- CN118236369A CN118236369A CN202410649279.XA CN202410649279A CN118236369A CN 118236369 A CN118236369 A CN 118236369A CN 202410649279 A CN202410649279 A CN 202410649279A CN 118236369 A CN118236369 A CN 118236369A
- Authority
- CN
- China
- Prior art keywords
- pyrroltinib
- iii
- tyrosine kinase
- kinase inhibitor
- atractylenolide
- 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
- FBMORZZOJSDNRQ-GLQYFDAESA-N Atractylenolide III Chemical compound C=C([C@@H]1C2)CCC[C@]1(C)C[C@@]1(O)C2=C(C)C(=O)O1 FBMORZZOJSDNRQ-GLQYFDAESA-N 0.000 title claims abstract description 198
- FBMORZZOJSDNRQ-UHFFFAOYSA-N Demethoxy,B,HCl-Adriamycin Natural products C1C2C(=C)CCCC2(C)CC2(O)C1=C(C)C(=O)O2 FBMORZZOJSDNRQ-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 206010012735 Diarrhoea Diseases 0.000 title claims abstract description 90
- OAXKIRPCKWQWOQ-UHFFFAOYSA-N atractylenolide III Natural products CC1=C2CC3C(CCCC3=C)CC2(O)OC1=O OAXKIRPCKWQWOQ-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 229940121358 tyrosine kinase inhibitor Drugs 0.000 title claims abstract description 47
- 239000005483 tyrosine kinase inhibitor Substances 0.000 title claims abstract description 42
- 150000004917 tyrosine kinase inhibitor derivatives Chemical class 0.000 title claims abstract description 31
- 230000000968 intestinal effect Effects 0.000 claims abstract description 33
- 239000003814 drug Substances 0.000 claims abstract description 32
- 108010079245 Cystic Fibrosis Transmembrane Conductance Regulator Proteins 0.000 claims abstract description 23
- 102100036009 5'-AMP-activated protein kinase catalytic subunit alpha-2 Human genes 0.000 claims abstract description 19
- 101000783681 Homo sapiens 5'-AMP-activated protein kinase catalytic subunit alpha-2 Proteins 0.000 claims abstract description 19
- 150000002596 lactones Chemical class 0.000 claims abstract description 15
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims abstract description 14
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 13
- SMEROWZSTRWXGI-HVATVPOCSA-N lithocholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 SMEROWZSTRWXGI-HVATVPOCSA-N 0.000 claims abstract description 13
- SMEROWZSTRWXGI-UHFFFAOYSA-N Lithocholsaeure Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 SMEROWZSTRWXGI-UHFFFAOYSA-N 0.000 claims abstract description 12
- 230000028327 secretion Effects 0.000 claims abstract description 8
- 230000026731 phosphorylation Effects 0.000 claims abstract description 6
- 238000006366 phosphorylation reaction Methods 0.000 claims abstract description 6
- 230000000694 effects Effects 0.000 claims description 29
- 102000012605 Cystic Fibrosis Transmembrane Conductance Regulator Human genes 0.000 claims description 20
- 239000002207 metabolite Substances 0.000 claims description 17
- 230000002829 reductive effect Effects 0.000 claims description 11
- 239000002246 antineoplastic agent Substances 0.000 claims description 10
- 229940041181 antineoplastic drug Drugs 0.000 claims description 10
- 241000193403 Clostridium Species 0.000 claims description 9
- 241000132012 Atractylodes Species 0.000 claims description 8
- 230000006378 damage Effects 0.000 claims description 8
- 230000001105 regulatory effect Effects 0.000 claims description 7
- 229960004641 rituximab Drugs 0.000 claims description 7
- 239000005411 L01XE02 - Gefitinib Substances 0.000 claims description 6
- 239000005551 L01XE03 - Erlotinib Substances 0.000 claims description 6
- 239000002136 L01XE07 - Lapatinib Substances 0.000 claims description 6
- 229960001686 afatinib Drugs 0.000 claims description 6
- ULXXDDBFHOBEHA-CWDCEQMOSA-N afatinib Chemical compound N1=CN=C2C=C(O[C@@H]3COCC3)C(NC(=O)/C=C/CN(C)C)=CC2=C1NC1=CC=C(F)C(Cl)=C1 ULXXDDBFHOBEHA-CWDCEQMOSA-N 0.000 claims description 6
- 230000009286 beneficial effect Effects 0.000 claims description 6
- 229960001433 erlotinib Drugs 0.000 claims description 6
- AAKJLRGGTJKAMG-UHFFFAOYSA-N erlotinib Chemical compound C=12C=C(OCCOC)C(OCCOC)=CC2=NC=NC=1NC1=CC=CC(C#C)=C1 AAKJLRGGTJKAMG-UHFFFAOYSA-N 0.000 claims description 6
- 229960002584 gefitinib Drugs 0.000 claims description 6
- XGALLCVXEZPNRQ-UHFFFAOYSA-N gefitinib Chemical compound C=12C=C(OCCCN3CCOCC3)C(OC)=CC2=NC=NC=1NC1=CC=C(F)C(Cl)=C1 XGALLCVXEZPNRQ-UHFFFAOYSA-N 0.000 claims description 6
- 229960004891 lapatinib Drugs 0.000 claims description 6
- BCFGMOOMADDAQU-UHFFFAOYSA-N lapatinib Chemical compound O1C(CNCCS(=O)(=O)C)=CC=C1C1=CC=C(N=CN=C2NC=3C=C(Cl)C(OCC=4C=C(F)C=CC=4)=CC=3)C2=C1 BCFGMOOMADDAQU-UHFFFAOYSA-N 0.000 claims description 6
- 229960000575 trastuzumab Drugs 0.000 claims description 6
- 230000002265 prevention Effects 0.000 claims description 5
- 208000000260 Warts Diseases 0.000 claims description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- 201000010153 skin papilloma Diseases 0.000 claims description 4
- 210000000813 small intestine Anatomy 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 230000002411 adverse Effects 0.000 claims description 3
- 230000007423 decrease Effects 0.000 claims description 3
- 230000001404 mediated effect Effects 0.000 claims description 3
- 210000004400 mucous membrane Anatomy 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 24
- 229940079593 drug Drugs 0.000 abstract description 20
- 102000004169 proteins and genes Human genes 0.000 abstract description 15
- 206010028980 Neoplasm Diseases 0.000 abstract description 6
- 230000007246 mechanism Effects 0.000 abstract description 5
- 241000092665 Atractylodes macrocephala Species 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- 230000003334 potential effect Effects 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 2
- 102000008371 intracellularly ATP-gated chloride channel activity proteins Human genes 0.000 abstract 1
- 241000700159 Rattus Species 0.000 description 48
- 241000894007 species Species 0.000 description 20
- 210000001519 tissue Anatomy 0.000 description 20
- 238000004458 analytical method Methods 0.000 description 19
- 239000000203 mixture Substances 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- 238000000034 method Methods 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 8
- 210000003734 kidney Anatomy 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 210000000952 spleen Anatomy 0.000 description 6
- 230000008485 antagonism Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 239000007850 fluorescent dye Substances 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 230000002195 synergetic effect Effects 0.000 description 5
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 4
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 4
- 210000001072 colon Anatomy 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 4
- DSLLHVISNOIYHR-UHFFFAOYSA-M ethyl 2-(6-methoxyquinolin-1-ium-1-yl)acetate;bromide Chemical compound [Br-].COC1=CC=C2[N+](CC(=O)OCC)=CC=CC2=C1 DSLLHVISNOIYHR-UHFFFAOYSA-M 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 238000004393 prognosis Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241001430149 Clostridiaceae Species 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 3
- 239000006180 TBST buffer Substances 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 230000030833 cell death Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 231100000915 pathological change Toxicity 0.000 description 3
- 230000036285 pathological change Effects 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- PNRKIXSZTUCIJN-UHFFFAOYSA-N 3-Hydroxybenzyl alcohol glucoside Chemical compound OC1C(O)C(O)C(CO)OC1OCC1=CC=CC(O)=C1 PNRKIXSZTUCIJN-UHFFFAOYSA-N 0.000 description 2
- 241000702460 Akkermansia Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 208000031648 Body Weight Changes Diseases 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 239000007995 HEPES buffer Substances 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 239000002033 PVDF binder Substances 0.000 description 2
- 229940122907 Phosphatase inhibitor Drugs 0.000 description 2
- 241000700157 Rattus norvegicus Species 0.000 description 2
- 241001261005 Verrucomicrobia Species 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 239000007640 basal medium Substances 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000004579 body weight change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 239000005414 inactive ingredient Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 210000004347 intestinal mucosa Anatomy 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- RDOIQAHITMMDAJ-UHFFFAOYSA-N loperamide Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(C(=O)N(C)C)CCN(CC1)CCC1(O)C1=CC=C(Cl)C=C1 RDOIQAHITMMDAJ-UHFFFAOYSA-N 0.000 description 2
- 229960001571 loperamide Drugs 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 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
- 238000013507 mapping Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000004877 mucosa Anatomy 0.000 description 2
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- 238000000513 principal component analysis Methods 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- FBVVGPMIPAZFAW-RZVRUWJTSA-N (2S)-pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1.OC(=O)[C@@H]1CCCN1 FBVVGPMIPAZFAW-RZVRUWJTSA-N 0.000 description 1
- SADXACCFNXBCFY-IYNHSRRRSA-N (e)-n-[4-[3-chloro-4-(pyridin-2-ylmethoxy)anilino]-3-cyano-7-ethoxyquinolin-6-yl]-3-[(2r)-1-methylpyrrolidin-2-yl]prop-2-enamide Chemical compound C=12C=C(NC(=O)\C=C\[C@@H]3N(CCC3)C)C(OCC)=CC2=NC=C(C#N)C=1NC(C=C1Cl)=CC=C1OCC1=CC=CC=N1 SADXACCFNXBCFY-IYNHSRRRSA-N 0.000 description 1
- HZYZHUZIWFRBNU-UHFFFAOYSA-N 4-hydroxybutanoic acid Chemical compound OCCCC(O)=O.OCCCC(O)=O HZYZHUZIWFRBNU-UHFFFAOYSA-N 0.000 description 1
- LOLRKBVIGZVBHY-UHFFFAOYSA-N 5-hydroxypentanoic acid Chemical compound OCCCCC(=O)O.OCCCCC(=O)O LOLRKBVIGZVBHY-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 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 1
- 241000606125 Bacteroides Species 0.000 description 1
- 241000605059 Bacteroidetes Species 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 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 1
- 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 1
- 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 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 206010014418 Electrolyte imbalance Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000609971 Erysipelotrichaceae Species 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 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 1
- 208000017891 HER2 positive breast carcinoma Diseases 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 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 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 241000605861 Prevotella Species 0.000 description 1
- 241000692844 Prevotellaceae Species 0.000 description 1
- 244000061632 Prinsepia uniflora Species 0.000 description 1
- 235000008997 Prinsepia uniflora Nutrition 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241000934136 Verruca Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 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 1
- 230000002457 bidirectional effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 235000012241 calcium silicate Nutrition 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
- 238000004364 calculation method Methods 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000000973 chemotherapeutic effect Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 230000007012 clinical effect Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 231100001159 controllable toxicity Toxicity 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000002790 cross-validation Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000002716 delivery method Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000010201 enrichment analysis Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 235000013861 fat-free Nutrition 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 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
- 230000037353 metabolic pathway Effects 0.000 description 1
- 238000002705 metabolomic analysis Methods 0.000 description 1
- 230000001431 metabolomic effect Effects 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 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
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 229940075576 pyrotinib Drugs 0.000 description 1
- 238000012207 quantitative assay Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 210000002460 smooth muscle Anatomy 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000000528 statistical test Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 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
- 238000012385 systemic delivery Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 108700043108 vasectrin III Proteins 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/12—Antidiarrhoeals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of biological medicines, and particularly relates to application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitors. Specifically, the research shows that the atractylenolide III can relieve diarrhea caused by the pyrroltinib, and the anti-tumor effect of the pyrroltinib is not affected. Atractylodes macrocephala lactone III can inhibit chloride ion secretion caused by pyrroltinib, and a potential action mechanism of the atractylenolide III is probably to increase AMPK phosphorylation and reduce protein expression of CFTR. Meanwhile, the atractylenolide III can relieve diarrhea caused by the pyrroltinib by adjusting the structure of intestinal flora and increasing the content of the lithocholic acid. The invention provides a new traditional Chinese medicine target for treating the drug diarrhea of the tyrosine kinase inhibitor such as the pyrroltinib, widens the clinical application and development of the atractylenolide III, further carries forward and plays the attenuation function of the traditional Chinese medicine in tumor treatment, thereby having good practical application value.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitors.
Background
Tyrosine kinase inhibitors (tyrosine kinase inhibitor, TKI) are the most widely used small molecule oral targeted drugs in clinic, and achieve ideal clinical effects in various solid tumors. Compared with the traditional chemotherapy scheme, the TKI targeting drug has the advantages of oral administration route, high target selectivity, obviously prolonged survival prognosis of sensitive patients and the like. TKI is typically one of the medicines, namely pyrroltinib, is a new medicine of 1.1 class which is originally researched in China, is widely used for clinical treatment of sensitive tumors such as lung cancer, breast cancer and the like, and has better curative effect than similar medicines. Pyrroltinib can significantly extend median progression-free survival (median progression-free survival, mPFS) and median total survival (median overall survival, mOS) of HER2 positive breast and lung cancer patients.
Although the clinical efficacy of pyrroltinib has been widely accepted, its diarrhea side effects are not negligible. The overall diarrhea incidence of the pyrroltinib is 95%, and the incidence of diarrhea (affecting life self-care or requiring hospitalization) of grade 3 or more is 40%. Diarrhea severely affects the quality of life of the patient, reduces drug compliance, induces electrolyte imbalance, and may be life threatening to serious. In addition, it places an economic burden on the patient. At present, no specific medicine for treating diarrhea caused by the pyrroltinib exists clinically, medicines such as loperamide and the like for treating diarrhea related to a chemotherapeutic medicine are still used, but the dosage and the administration time of the loperamide are not easy to grasp, and secondary side reactions such as constipation and bellyache are easy to cause due to the characteristics of inhibiting the contraction of intestinal smooth muscle and reducing intestinal peristalsis, so that the life quality of patients is further reduced. In order to solve the clinical problem, it is important to find a safe and effective drug which is suitable for long-term administration.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention provides application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitors. Specifically, the research shows that the atractylenolide III can relieve diarrhea caused by the tyrosine kinase inhibitor pyrroltinib, and the anti-tumor effect of the pyrroltinib is not affected. Provides a new traditional Chinese medicine target for the treatment of the diarrhea of TKI medicines such as pyrroltinib, widens the clinical application and development of the atractylenolide III, and further develops and plays the attenuation function of the traditional Chinese medicine in tumor treatment. Based on the above-described results, the present invention has been completed.
Specifically, the invention relates to the following technical scheme:
in a first aspect, the invention provides application of atractylenolide III in preparing a medicament for preventing and treating diarrhea caused by tyrosine kinase inhibitors.
Wherein the tyrosine kinase inhibitor includes, but is not limited to, trastuzumab, rituximab, lapatinib, lenatinib, pyrroltinib, afatinib, gefitinib, and erlotinib, and in one embodiment of the invention, the tyrosine kinase inhibitor is pyrroltinib.
Specifically, the prevention and treatment of diarrhea caused by tyrosine kinase inhibitors is specifically prevention and/or treatment of diarrhea caused by pyrroltinib, and the anti-tumor effect of pyrroltinib is not affected.
More specifically, the prevention and treatment of diarrhea caused by tyrosine kinase inhibitors is shown by:
(a) The damage of the mucous membrane of the small intestine caused by the pyrroltinib is reduced;
(b) Inhibiting chloride ion secretion caused by pyrroltinib;
(c) Regulating the intestinal flora disorder caused by pyrroltinib;
(d) Increasing the content of the beneficial metabolite lithocholic acid.
Wherein (b) specifically comprises: bighead atractylodes lactone III promotes AMPK phosphorylation and/or inhibits CFTR protein expression.
The specific steps in (c) are as follows: bighead lactone III increases the levels of the beneficial flora wart and Ackermansia, decreases the levels of the adverse flora Clostridium (Clostridiaceae/Clostridium) to alleviate pyrrole-induced diarrhea.
In a second aspect of the invention, the application of the tyrosine kinase inhibitor combined with the atractylenolide III in preparing anti-tumor drugs is provided.
Specifically, the antitumor drug is specifically expressed as follows: exerts the anti-tumor effect of the tyrosine kinase inhibitor and prevents and/or treats diarrhea side effects mediated by the tyrosine kinase inhibitor.
The mass ratio of the tyrosine kinase inhibitor to the atractylenolide III is 16:1-3, such as 16:1, 16:2 and 16:3; or the mol ratio of the tyrosine kinase inhibitor to the atractylenolide III is 1-4:1000-25000, such as 1:1000, 1:2500, 1:6250.
Wherein the tyrosine kinase inhibitor includes, but is not limited to, trastuzumab, rituximab, lapatinib, lenatinib, pyrroltinib, afatinib, gefitinib, and erlotinib, and in one embodiment of the invention, the tyrosine kinase inhibitor is pyrroltinib.
In a third aspect of the present invention, there is provided an antitumor drug comprising at least a tyrosine kinase inhibitor and atractylenolide III as active ingredients.
Wherein the mass ratio of the tyrosine kinase inhibitor to the atractylenolide III is 16:1-3, such as 16:1, 16:2 and 16:3; or the mol ratio of the tyrosine kinase inhibitor to the atractylenolide III is 1-4:1000-25000, such as 1:1000, 1:2500, 1:6250.
The tyrosine kinase inhibitors include, but are not limited to, trastuzumab, rituximab, lapatinib, lenatinib, pyrroltinib, afatinib, gefitinib, and erlotinib, and in one embodiment of the invention, the tyrosine kinase inhibitor is pyrroltinib.
Specifically, the antitumor drug can effectively relieve diarrhea side effects mediated by the pyrroltinib besides effectively playing the antitumor effect of the pyrroltinib.
According to the invention, the antitumor drug may further comprise at least one pharmaceutically inactive ingredient.
The pharmaceutically inactive ingredients may be carriers, excipients, diluents and the like which are generally used in pharmacy. Further, the composition can be formulated into various dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, sprays, etc., for oral administration, external use, suppositories, and sterile injectable solutions according to a usual method.
The non-pharmaceutically active ingredients, such as carriers, excipients and diluents, which may be included, are well known in the art and can be determined by one of ordinary skill in the art to meet clinical criteria.
In yet another embodiment of the present invention, the carriers, excipients and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. The present invention is not particularly limited herein.
In yet another embodiment of the invention, the medicament of the invention may be administered to the body in a known manner. Such as systemic delivery via veins. Administration may optionally be via intravenous, transdermal, intranasal, oral, mucosal, or other delivery methods. Such administration may be via single or multiple doses. It will be appreciated by those skilled in the art that the actual dosage to be administered in the present invention may vary greatly depending on a variety of factors, such as the target cell, the type of organism or tissue thereof, the general condition of the subject to be treated, the route of administration, the mode of administration, and the like.
In a fourth aspect of the invention, there is provided a method of tumour therapy, the method comprising administering to a subject an effective amount of an anti-tumour agent as described above.
An "effective amount" as used herein refers to an amount of an active compound or agent, including a compound of the present invention, that is capable of eliciting a biological or medical response in a tissue system, animal or human that is sought by a researcher, veterinarian, medical doctor or other medical personnel, which includes alleviation or partial alleviation of the symptoms of the disease, syndrome, condition or disorder being treated.
In a further specific embodiment of the invention, the mass ratio of the tyrosine kinase inhibitor to the atractylenolide III in the anti-tumor drug is 16:1-3, such as 16:1, 16:2 and 16:3; or the mol ratio of the tyrosine kinase inhibitor to the atractylenolide III is 1-4:1000-25000, such as 1:1000, 1:2500, 1:6250.
The beneficial technical effects of one or more of the technical schemes are as follows:
experiments prove that the technical scheme can relieve diarrhea caused by the pyrroltinib and does not influence the anti-tumor effect of the pyrroltinib. Atractylodes macrocephala lactone III can inhibit chloride ion secretion caused by pyrroltinib, and a potential action mechanism of the atractylenolide III is probably to increase AMPK phosphorylation and reduce protein expression of CFTR. Meanwhile, the atractylenolide III can relieve diarrhea caused by the pyrroltinib by adjusting the structure of intestinal flora and increasing the content of the lithocholic acid.
In a word, the technical scheme provides a new traditional Chinese medicine target for treating the diarrhea of TKI medicines such as pyrroltinib, widens the clinical application and development of the atractylenolide III, further carries forward and plays the attenuation function of the traditional Chinese medicine in tumor treatment, and therefore has good practical application value.
Drawings
FIG. 1 is a diagram showing the overall experimental design of an animal in an embodiment of the present invention.
FIG. 2 is a graph showing the evaluation result of diarrhea in rats according to the embodiment of the present invention; (A) - (B) respectively showing the diarrhea incidence rate and diarrhea scoring result of rats with diarrhea caused by the intervention of different doses of atractylenolide III; (B) The result : C vs P,p＜0.0001;C vs P+Ⅲ (H),p＜0.0001;C vs P+Ⅲ (M),p＜0.0001;C vs P+Ⅲ (L),p＜0.0001;P vs P+Ⅲ (H),p＜0.0001;P vs P+Ⅲ (M),p＜0.0001;P vs P+Ⅲ (L),p=0.0034;P+Ⅲ (H) vs P+Ⅲ (M),p=0.2561;P+Ⅲ (H) vs P+Ⅲ (L),p＜0.0001;P+Ⅲ (M) vs P+Ⅲ (L),p=0.0153;（C）-（D） of the medium-significant difference analysis shows the incidence rate of diarrhea and the result of diarrhea scoring of rats with diarrhea caused by the intervention of high-dose atractylenolide III in pyrroltinib respectively; (D) The medium significance analysis result ：C vs Ⅲ (H),p=0.999;C vs P+Ⅲ (H),p＜0.0001;C vs P,p＜0.0001;Ⅲ (H)vs P+Ⅲ (H),p＜0.0001;Ⅲ (H)vs P,p＜0.0001;P+Ⅲ (H)vs P,p＜0.0001;C represents a control group; p represents a pyrroltinib group; III (H, M, L) represents the group of atractylenolides III (high, medium, low doses); III (H, M, L) +P represents the combination of atractylenolide III (high, medium, low dose) with pyrroltinib group.
FIG. 3 shows the results of body weight change, diarrhea index, spleen index and kidney index of rats in the examples of the present invention; (A) represents a change in body weight of the rat; (B) represents diarrhea index results for rats; (C) shows the spleen index results of rats. (D) represents kidney index results of rats; c represents a control group; p represents a pyrroltinib group; III (H) represents the high dose group of atractylenolide III; III (H) +P represents the high dose combination of atractylenolide III with the pyrroltinib group; * Represents p < 0.05; * Represents p < 0.01; * Represents p < 0.001; * P < 0.0001; ns represents p.gtoreq.0.05.
FIG. 4 shows the H & E staining results of rat intestinal tissue in the examples of the present invention; (A) - (D) sequentially represents the H & E staining results of the colon tissue of the rat treated by the control group, the high-dose atractylenolide group III, the pyrroltinib group and the high-dose atractylenolide group III combined with the pyrroltinib group; (E) - (H) shows the H & E staining results of the rat small intestine tissue treated by the control group, the high-dose atractylenolide III group, the pyrroltinib group and the high-dose atractylenolide III group combined with the pyrroltinib group in sequence.
FIG. 5 shows the results of MAQE chloride fluorescent probes after interfering T84 cells with pyrroltinib and atractylenolide III in the examples of the present invention; (A) Is a fluorescence diagram of the dry prognosis of the control group, the pyrroltinib group, the atractylenolide III group and the pyrroltinib combined atractylenolide III group; (B) The fluorescence result statistical analysis of the dry prognosis of the control group, the pyrroltinib group, the atractylenolide III group and the pyrroltinib combined atractylenolide III group; * Represents p < 0.05; * Represents p < 0.01.
FIG. 6 shows the effect of atractylenolide III on the AMPK/CFTR signal pathway in an embodiment of the present invention; (A) - (B) mRNA expression of AMPK and CFTR genes, respectively; (C) Gel electrophoresis patterns of AMPK (alpha), p-AMPK (alpha) and CFTR proteins; (D) p-AMPK/AMPK protein expression; (E) CFTR protein expression; * Represents p < 0.05; * Represents p < 0.01; * Represents p < 0.001; * P < 0.0001; ns represents p.gtoreq.0.05.
FIG. 7 shows the results of analysis of Alpha diversity and Beta diversity of intestinal flora in examples of the present invention; (A) is a box plot of Alpha diversity index; (B) a two-dimensional ordering diagram of Beta diversity PCoA analysis; each dot in the figure represents one sample, with the different colored dots indicating different samples; c represents a control group; p represents a pyrroltinib group; III represents the group of atractylenolides III and P_III represents the group of atractylenolides III in combination with pyrroltinib.
FIG. 8 is a heat map of the composition of a genus level species of dual clusters in an embodiment of the present invention; red represents that the abundance of the genus in this sample is higher than in other samples; blue represents the opposite meaning.
FIG. 9 is a bar graph of LDA effect values of marker species according to an embodiment of the present invention; the ordinate is the class units with significant differences between the groups, and the abscissa is the LDA effect value for each class unit.
FIG. 10 is a graph of Zi-Pi scattergrams of an intestinal flora species-associated network in accordance with an embodiment of the present invention.
FIG. 11 shows the results of rat metabolites in examples of the present invention. (A) - (B) PCA score plot in metabolite positive and negative ion mode, respectively; the abscissa PC1 represents the first principal component score value, and the ordinate PC2 represents the second principal component score value; points represent samples, circles represent 95% confidence intervals; (C) a violin diagram of the secondary differential metabolite lithocholic acid; the abscissa represents the group, and the ordinate represents the metabolite quantitative value; the stars between the two groups represent the significance of the difference between the two groups; * Represents p < 0.05; ns represents that p is greater than or equal to 0.05; c is a control group; p is a pyrroltinib group; III is the atractylenolide III group, and P_III is the atractylenolide III combined with the pyrroltinib group.
FIG. 12 shows the results of CompuSyn software analysis of atractylenolide III and pyrroltinib in the examples of the present invention; (A) Dose-inhibition curves for different concentrations of atractylenolide iii in combination with pyrroltinib; (B) is an isobologram of atractylenolide III and pyrroltinib; fa is the inhibition; p is a pyrroltinib group; III is the group III of atractylenolide; P+III is the group III of pyrroltinib combined with atractylenolide.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
The invention will be further illustrated with reference to specific examples, which are given for the purpose of illustration only and are not to be construed as limiting the invention. If experimental details are not specified in the examples, it is usually the case that the conditions are conventional or recommended by the sales company; materials, reagents and the like used in the examples were commercially available unless otherwise specified.
Examples
1. Materials and methods
1.1 General design of animal experiments
The study was conducted in strict compliance with the declaration of helsinki and had passed the laboratory animal ethics committee of the affiliated province of the first medical science, shandong (ethics lot number: no. 2023-146). Pyrroltinib was purchased from Jiangsu Hengrui medicine Co., ltd (6124230401 cs), and atractylenolide III was purchased from Dou Desai Tex Biotechnology Co., ltd (DSTDB 001602). SPF grade Wistar rats of about 5 weeks of age for 10 days of experiment （Lai J, Zhuo X, Yin K, et al. Potential mechanism of pyrotinib-induced diarrhea was explored by gut microbiome and ileum metabolomics.Anti-cancer Drugs. 2022）. were randomly divided into a control group (equal dose physiological saline), a pyrroltinib group (80 mg/kg/d), a largehead atractylodes rhizome lactone III low, medium and high doses (5 mg/kg/d, 10mg/kg/d, 15 mg/kg/d) in combination with a pyrroltinib group (80 mg/kg/d) and the like, each group being 6, in total, 54, all purchased from Vetong rituximab Co., ltd, by using the method of the pre-established pyrroltinib (80 mg/kg/d) Wistar rat diarrhea model of the subject group. Raising in Shandong university of first medical science animal research center, environmental condition: the temperature was 22.+ -. 1 ℃, the relative humidity was 50.+ -. 1% and the light/dark period was 12h/12h. Adaptive feeding was performed for 1 week before the experimental study was formally started. The mode of the combined group administration of the atractylenolide III with different dosages is that the atractylenolide III is given to the rat after half an hour. Finally, the efficacy was verified using the optimal dose, and the overall study design is shown in fig. 1. The diarrhea condition of the rats was observed daily (the diarrhea grade of the rats is shown in Table 1), the occurrence time and frequency of diarrhea was recorded, the general conditions of body weight, hair, etc. of the rats were recorded every two days, and diarrhea scoring was performed to evaluate the diarrhea index and the organ index. Diarrhea score = average of diarrhea scores for all animals of the group. Diarrhea index = runny bowel rate x average runny bowel grade. (loose stool rate=number of loose stools per animal (grade ii diarrhea and above)/total number of loose stools per animal; average loose stool grade=number of loose stools (grade ii diarrhea and above)/number of loose stools; number of loose stools=number of diarrhea grade x number of diarrhea grade). Organ index = weight of organ at time of sacrifice (g)/weight of rat (g). After the end of the experiment, the rats were sacrificed and rat spleen, kidney, intestinal tissue and rat feces were collected for subsequent experiments.
TABLE 1 grading of diarrhea in animals
1.2 Hematoxylin and eosin (hematoxylin and eosin, H & E) staining to assess pathological changes in intestinal tissue
The intestinal tissues of rats fixed for more than 24 hours are dehydrated. And after dehydration, adding paraffin, taking out after cooling and solidifying, cutting into slices, flattening the slices, floating on water at 40 ℃ of a tissue slice spreading machine, fishing out by using a glass slide, baking the slices in a 60 ℃ oven, taking out after the paraffin is melted, and carrying out H & E dyeing on the tissue slices. The prepared intestinal tissue H & E stained sections are placed under an integrated fluorescence microscopy imaging system microscope (BZ-X800E) to observe the damage degree, a tissue damage scoring system is used for scoring (see table 2), the scoring is carried out by two scoring staff, and finally the obtained scoring is added to represent pathological changes of intestinal tissues, wherein the higher the scoring is, the more obvious the pathological changes are.
Table 2 tissue injury scoring system
1.3 MQAE chloride ion fluorescent probe for detecting green fluorescence intensity in T84 cells
The effect of the drug on chloride ion secretion was evaluated using MQAE chloride ion fluorescent probes. Human colon cancer cell line T84 (Wahanprinomi Life technologies Co., ltd., CL-0229) in logarithmic growth phase was taken for digestion, and cells were resuspended in complete medium (DMEM/F-12:1 basal medium+10% FBS+1% penicillin-streptomycin double antibody) and plated in 24 well plates at a density of 5X 10 4 per well. After the cell growth state is good, adding the pyrroltinib and the atractylenolide III medicines respectively for intervention for 24 hours, washing the cell for 1-2 times by PBS, and incubating the cell in a Krebs-HEPES buffer solution containing 5mM at 37 ℃ for 30-60min in a dark place. After the incubation, cells were washed 5-6 times with Krebs-HEPES buffer, green fluorescence was captured using an integrated fluorescence microscopy imaging system microscope (BZ-X800E), and analyzed for fluorescence intensity using imageJ software.
1.4 RT-qPCR (reverse transcription-quantitative polymerase chain reaction) detection of mRNA (messenger ribonucleic acid) expression level of targeted gene
The experiment designs primers of rat-derived internal reference genes GADPH and target genes CFTR and AMPK, synthesized by the company of the department of Optimus of the family of the Prinsepia, the specific primer sequences are shown in Table 3. RNA from rat intestinal tissue was extracted using a high purity RNA kit (Magen, md. 022). cDNA was obtained using a reverse transcription kit (Mona Biol, MR 05001S/M). The reverse transcription solution system was 20 μl: 10 μl of RNA reagent for removing genomic DNA contamination, monScript ™ 5 × RTIII All-in-One Mix 4 μl, and DEPC water 6 μl. qPCR experiments were performed using qPCR kit (Ebolac, RK 21203). The qPCR single reaction system was 10. Mu.l: 2X MonAmp ™ SYBR cube GREEN QPCR Mix 5. Mu.l, target gene forward primer (F) 0.3. Mu.l, target gene reverse primer (R) 0.3. Mu.l, cDNA 1. Mu.l, DEPC water 3.4. Mu.l. The whole experimental procedure was performed on ice. Setting a reaction program according to the steps of the specification, confirming amplification and melting curves after finishing, respectively normalizing the relative level of mRNA expression of a target gene and a reference gene GADPH, and analyzing the relative expression quantity of the target gene by adopting a 2 （-ΔΔCT） method.
TABLE 3 primer sequences used in this example
1.5 Western Blot detection of key protein expression level of targeted gene
Lysates were prepared at the ratio RIPA: PMSF: phosphatase inhibitor a solution: phosphatase inhibitor B solution=98:1:0.5:0.5 to extract proteins from rat intestinal tissue, and the whole process was performed on ice. Protein concentration was determined using BCA protein quantitative assay kit (Thermo FISHER SCIENTIFIC, 23227) to assess electrophoretic loading of tissue proteins. Electrophoresis was performed by SDS polyacrylamide gel, and after the completion of electrophoresis, the gel was transferred to a PVDF membrane having a pore size of 0.45. Mu.m. The membranes were blocked in 5% nonfat dry milk blocking solution/1% BSA blocking solution for 1 hour on a low speed shaker. After the end, the shaker was washed 3 times with TBST solution for 8 min/time. After the end, the destination bands are respectively set at 1:1000 CFTR antibody (SANTA CRUZ Co., sc-376683), 1: AMPK alpha antibody (CST, 2532) 1000, 1:1000 p-AMPK alpha antibody (CST company, 5759) and 1:1000 beta-actin antibodies (Abways, no. AB 0035) were incubated overnight at 4℃and washed 3 times with TBST solution for 8 min/time. After the end, the PVDF film was put into 1:5000 horseradish peroxidase-conjugated goat anti-mouse or goat anti-rabbit IgG secondary antibodies (Abways, anti-mouse No. AB0102, anti-rabbit No. AB 0101) were incubated at room temperature for 1 hour, and after incubation, the TBST solution was washed 3 times for 8 minutes/time. After completion, the bands were developed using ECL developer (Tanon, no: 1805001W/B) and the grey values of the bands were quantified using imageJ software.
1.6 Evaluation of intestinal flora by 16S rRNA
Sequencing library was prepared using TruSeq nano DNA LT library preparation kit from Illumina, inc., and the sequence of the V3-V4 region of the 16S rRNA gene was detected. The operational taxa occurring in 50% or more of the stool samples were identified as core operational taxa. Seven indices, chao1, observed species, shannon, simpson, faith's PD, pielou ' S EVENNESS and Good's coverage, were used to assess Alpha diversity of the intestinal flora. The Chao1 and Observed species indices represent richness, shannon and Simpson indices represent diversity, faith's PD index represents evolutionarily based diversity, pielou ' S EVENNESS index represents uniformity, and Good's coverage index represents coverage. Specific calculation methods are detailed in :http://scikit-bio.org/docs/latest/generated/skbio.diversity.alpha.html#module-skbio.diversity.alpha. for evaluation of Beta diversity of intestinal flora using the principle coordinate analysis (PRINCIPAL COORDINATES ANALYSIS, PCoA) method. Analysis was performed using the class-level taxonomic composition as a target and heat maps and LEfSe (LDA EFFECT size) analyses were drawn with the abundance data of the first 50 genera of average abundance, resulting in species differences and marker species. Nodes in the network are divided into four parts, namely peripheral nodes, connection nodes, module center points and network center points, by using Zi and Pi score values (the Zi value refers to intra-module connectivity, and the Pi refers to inter-module connectivity), and the nodes are used for searching key species. The prediction of functional potential of the flora was performed using KEGG (KEGG PATHWAY database, https:// www.kegg.jp /).
1.7 Metabonomics assessment of Metabolic product Condition
The present study uses principal component analysis (PRINCIPAL COMPONENT ANALYSIS, PCA) to assess the status of metabolites. PCA refers to that metabolite variables are linearly combined according to certain weights to generate new characteristic variables, and each group of data is classified through main new variables (main components). The closer the sample distribution points are, the closer the composition and concentration of the samples are; conversely, the greater the difference. Cross-validation of the model refers primarily to the R2X parameter, indicating the model's intelligibility (R2 is better than 0.5). Performing data scaling on the secondary differential metabolite matrix data by adopting R PHEATMAP program package, performing bidirectional clustering on samples and differential metabolites, drawing a clustering heat map, performing statistical test analysis on the correlation significance of the metabolites, selecting significance P <0.05 as the significance correlation, and finally drawing the overall distribution condition of the data by using a violin map. Finally, KEGG pathway enrichment analysis and topology analysis were performed on the differential metabolite list using MetaboAnalyst (https:// www.metaboanalyst.ca).
1.8 CCK8 method and CompuSyn software to evaluate the effect of atractylenolide III on the anti-tumor effect of pyrroltinib
HER2 (+) breast cancer cell line BT-474 (Wohunorace life technologies Co., ltd., CL-0040) in logarithmic growth phase was digested, cell suspension was prepared with complete medium (RPMI-1640 basal medium +10. Mu.g/ml Insulin +2mM L-glutamine +20% FBS +1% P/S), inoculated into 96-well plate at cell number of 3000-5000/well, and cultured in cell incubator (temperature 37 ℃, humidity 90%,5% CO 2) for 24 hours. The experiment is divided into a control group, a pyrroltinib group, a group of atractylenolide III with different concentration gradients (the latter concentration is 10 times of the former concentration) and a group of atractylenolide III combined with pyrroltinib, wherein each group is provided with 5 compound holes. After 24h of drug intervention, 10 mu L of CCK-8 reagent is added to each hole, incubated at 37 ℃ in a dark place for 30-60min, and the absorbance (OD value) is measured at 450nm of an enzyme-labeled instrument. Cell death rate was calculated: cell death (%) =100- (drug group OD value-standard group OD value)/(control group OD value-standard group OD value) ×100. And obtaining a metering-reaction curve of the atractylenolide III and the pyrroltinib according to the cell death rate and CompuSyn software analysis. The data points below the curves suggest that there is a synergy between the drugs; data points on the curves indicate that the medicines belong to superposition; data points above the curves indicate antagonism between drugs. Or to obtain the joint index CI value: a very strong synergistic effect of < 0.1; 0.1-0.3 strong synergistic effect; 0.3-0.7 synergistic effect; a moderate synergistic effect of 0.7-0.85; a slight synergistic effect of 0.85-0.9; 0.9-1.1 additive effect; 1.1-1.2 mild antagonism; 1.2-1.45 moderate antagonism; 1.45-3.3 antagonism; 3.3-10 strong antagonistic effect; > 10 extremely strong antagonism.
1.9 Statistical analysis and mapping of data
Analysis and mapping were performed using GRAPHPAD PRISM 9.0.0 software, the differences between the two groups were compared using T-test, and the differences between the groups were compared using one-way anova. When p <0.05, it is statistically significant.
2. Results
2.1 Effect of atractylenolide III on Zygosaponin diarrhea rats
In order to investigate the effect of atractylenolide III on rats with diarrhea caused by pyrroltinib, the present study was conducted using a model of diarrhea in rats of pyrroltinib (80 mg/kg/d) that had been constructed earlier in the subject group. The results show that rats of the pyrroltinib group and the largehead atractylodes rhizome lactone III combined with the pyrroltinib group with different dosages all have diarrhea with different degrees compared with the control group; compared with the pyrroltinib group, after the different doses of the atractylenolide III are combined with the pyrroltinib, the time for the first diarrhea of the rat to occur is delayed, the diarrhea degree and the diarrhea score are reduced, the curative effect of the high dose of the atractylenolide III group (15 mg/kg/d) is more obvious, and the difference has statistical significance, as shown in (A) - (B) in fig. 2. In order to further verify the effect of high dose of atractylenolide III group (15 mg/kg/D) on diarrhea caused by pyrroltinib, the study was conducted with atractylenolide III (15 mg/kg/D) and it was found that atractylenolide III alone did not cause diarrhea or other adverse reactions in rats, and the effect after combination with pyrroltinib was consistent with the above, as shown in (C) - (D) of FIG. 2.
2.2 Effect of atractylenolide III on weight of rats suffering from diarrhea caused by pyrroltinib
Evaluation of body weight, diarrhea index, spleen index and kidney index of rats of each group after dry prognosis of atractylenolide III (15 mg/kg/d) showed that: compared with the control group, the body weight change and diarrhea index of rats in the atractylenolide III group have no obvious difference, and the spleen index and the kidney index of the rats are slightly increased; compared with the control group, the atractylenolide III group and the atractylenolide III combined with the pyrroltinib, the weight gain of rats in the pyrroltinib group is slow or obviously reduced, the diarrhea index is obviously increased, the spleen index and the kidney index of the rats are reduced, and the difference has statistical significance, as shown in (A) - (D) in figure 3.
2.3 Effect of atractylenolide III on intestinal tissue in rats
To investigate the effect of atractylenolide III on rat intestinal tissue, hematoxylin and eosin (hematoxylin and eosin, H & E) staining was performed on rat small intestinal tissue and colon tissue. The results show that the atractylenolide III (15 mg/kg/d) and the control group have similar results, the pyrroltinib group can cause the damage of the small intestinal mucosa and does not cause the damage of the colon mucosa, and the atractylenolide III (15 mg/kg/d) combined with the pyrroltinib group has lighter damage of the small intestinal mucosa and no damage of the colon mucosa, as shown in (A) - (H) in fig. 4. These results suggest that the atractylenolide III (15 mg/kg/d) used in this experiment is in a range of controllable toxicity and can reduce the damage to the mucous membrane of the small intestine caused by pyrroltinib.
2.4 Effect of pyrroltinib and atractylenolide iii on green fluorescence intensity in T84 cells
And detecting green fluorescence in the T84 cells by adopting an MQAE chloride ion fluorescent probe to evaluate the influence of the pyrroltinib and the atractylenolide III on chloride ions. The result shows that compared with the control group, the green fluorescence of the pyrroltinib group is obviously enhanced, and the green fluorescence of the atractylenolide III group is obviously reduced; the green fluorescence of the atractylenolide III in combination with the pyrroltinib group was significantly reduced compared to that of the pyrroltinib group, as shown in FIGS. 5 (A) - (B).
2.5 Effect of Bighead atractylodes lactone III on AMPK/CFTR Signal pathway
To investigate the potential mechanism of action of atractylenolide III in relieving diarrhea in rats caused by pyrroltinib, rat intestinal tissues were used to detect mRNA expression of AMPK, CFTR and protein expression of AMPK, p-AMPK (alpha) and CFTR. RT-qPCR results show that compared with a control group, the mRNA of AMPK is down-regulated in a pyrroltinib group, the mRNA of CFTR is up-regulated in a atractylenolide III group (p is more than 0.05), and the mRNA of the CFTR is down-regulated in the pyrroltinib group (p is less than 0.05); compared with the pyrroltinib group, after the combined use of the atractylenolide III, the mRNA of AMPK is down-regulated (p is more than 0.05), the mRNA of CFTR is down-regulated (p is less than 0.05), and the mRNA is shown in (A) - (B) in fig. 6. This result suggests that atractylenolide III down-regulates the mRNA of CFTR. Western Blot results show that compared with a control group, the expression of p-AMPK (alpha) protein in the pyrroltinib group is reduced, the expression of the atractylenolide III group is increased, the expression of CFTR protein in the pyrroltinib group is increased, and the expression of the atractylenolide III group is reduced (p is less than 0.05); compared with the pyrroltinib group, the protein expression of p-AMPK (alpha) is increased and the protein expression of CFTR is reduced (p < 0.05) after the combined administration of the atractylenolide III, as shown in (C) - (E) in FIG. 6. This result demonstrates that atractylenolide III can reduce the protein expression of CFTR and increase the expression of p-AMPK (alpha) protein.
In combination with the short-circuit current and MQAE chloride fluorescent probe results, these results further suggest that pyrroltinib promotes chloride secretion by inhibiting AMPK phosphorylation, increasing protein expression of CFTR, while atractylenolide iii can inhibit chloride secretion by the opposite effect. In addition, the differential metabolites are subjected to KEGG enrichment, and the AMPK signal pathway is found to be involved in regulation, and the results comprehensively indicate that the AMPK/CFTR is a potential mechanism of the atractylenolide III for treating diarrhea caused by the pyrroltinib.
2.6 Effect of atractylenolide III on diarrhea rat intestinal flora caused by pyrroltinib
To investigate the effect of atractylenolide III on the intestinal flora of diarrhea rats caused by pyrroltinib, we performed 16S rRNA analysis.
2.6.1 Effect of atractylenolide III on Alpha diversity and Beta diversity of diarrhea rat intestinal flora caused by pyrroltinib
Seven indices, chao1, observed species, shannon, simpson, faith's PD, pielou ' S EVENNESS and Good's coverage, were used to assess Alpha diversity of the intestinal flora. The results indicate that the minimum inner circumferences Chao1 and observed_patterns index of the control group, the atractylenolide III group (15 mg/kg/d), the pyrroltinib group and the atractylenolide III group (15 mg/kg/d) combined with the pyrroltinib group are both more than 500, which indicates that the number of flora species of a sample in each group is more; the minimum inner periphery good_coverage indexes are all larger than 0.9985, which indicates that the microbial coverage rate of flora of samples in each group is high, the probability that new species in the samples are not detected is low, and the sequencing result reflects the real situation of the samples; the minimum inner circumference Shannon index is larger than 5, which indicates that the flora uncertainty and diversity of the samples in each group are large; the median Simpson index is greater than 0.9 and less than 1, which indicates that the sample in each group has high abundance of flora species; the minimum inner circumference Pielou' S EVENNESS had an index of greater than 0.5, indicating uniform population distribution of samples within each group, but no significant differences between groups, as shown in fig. 7 (a).
Beta diversity analysis was performed with PCoA. In the two-dimensional ordering diagram of Beta diversity PCoA analysis, the closer the projection distance of two points on the coordinate axis is, the more similar the community composition of the two samples in the corresponding dimension is. The results show that the projection distances of samples of the control group and the atractylenolide III (15 mg/kg/d) group on the coordinate axes are similar, which shows that the flora of the two groups has similar composition; the projection distance of the sample of the pyrroltinib group and the sample of the control group on the coordinate axis is far, which indicates that the composition of flora of the two groups is dissimilar; the projection distance of the atractylenolide III (15 mg/kg/d) combined with the sample of the pyrroltinib group and the other three groups on the coordinate axis is far, which shows that the composition of the flora of the other three groups is dissimilar, as shown in (B) in fig. 7. These results suggest that atractylenolide III alleviates diarrhea associated with intestinal flora caused by pyrroltinib.
2.6.2 Analysis of difference of species of rat intestinal flora caused by pyrroltinib by atractylenolide III and marker species
As shown in fig. 8, to compare species composition differences between samples, a generic horizontal species composition heat map of intestinal flora double-cluster at the first 50 of average abundance was plotted and LEfSe difference analysis was performed. Prevotella (Prevotellaceae) is the population with the highest abundance ratio of the control population; the verruca (Verrucomicrobia) and ackermansia (Akkermansia) are groups of lagehead atractylodes lactones with higher abundance ratios of group iii groups; lactobacillus (Erysipelotrichaceae) and Clostridium (Clostridium) are species with a higher abundance ratio of the pyrroltinib group of bacteria; clostridium (Clostridiaceae/Clostridium) is a group with a higher abundance ratio of atractylenolide iii in combination with the pyrroltinib group, but its abundance is not as high as that of pyrroltinib group, as these results shown in figure 9 suggest, bighead lactone III can increase the levels of the beneficial flora wart microzyme (Verrucomicrobia) and Ackermansia (Akkermansia), and decrease the levels of the adverse flora Clostridium (Clostridiaceae/Clostridium) to alleviate pyrrole-induced diarrhea.
2.6.3 Bighead atractylodes rhizome lactone III on rat intestinal flora species association network of diarrhea caused by pyrroltinib and KEGG analysis
The key species were searched using the Zi-Pi score values and found to be the key species for the Thick-walled bacteria phylum (Firmicutes) and the Bacteroides phylum (Bacteroidetes), as shown in FIG. 10. The KEGG secondary functional pathway abundance results of the flora show that the metabolic pathways of the flora are involved. These results suggest that the alleviation of diarrhea caused by pyrroltinib by atractylenolide III is associated with intestinal flora and its metabolites.
2.7 Effect of atractylenolide III on rat metabolites with diarrhea caused by pyrroltinib
To investigate the effect of atractylenolide III on the metabolic products of diarrhea rats caused by pyrroltinib, metabonomics analysis was performed. First, the PCA score plot in the positive and negative ion modes shows that the sample distribution points of each group used in the experiment at this time are closer, indicating that the composition and concentration of each group of samples are close, and can be used in the metabonomic analysis at this time, as shown in (a) - (B) in fig. 11. Further, the first six secondary differential metabolites in the control group, the atractylenolide III group (15 mg/kg/d), the pyrroltinib group, the atractylenolide III group (15 mg/kg/d) in combination with the pyrroltinib group were lithocholic acid (lithocholic acid, LCA), 4-hydroxybutyric acid (4-Hydroxybutanoic acid), benzaldehyde (Benzaldehyde), L-Proline (L-Proline), 5-hydroxyvaleric acid (5-Hydroxypentanoic acid) and 3-hydroxybenzyl alcohol glucoside (3-Hydroxybenzyl alcohol glucoside). Of these, lithocholic acid most significantly differed (p=0.0026, p < 0.01). As shown in fig. 11 (C), the content of lithocholic acid was increased in the atractylenolide iii group and decreased in the pyrroltinib group, compared to the control group; compared with the pyrroltinib group, the content of lithocholic acid after the combined administration of the atractylenolide III is increased. This result suggests that atractylenolide III relieves diarrhea caused by pyrroltinib by increasing the metabolite lithocholic acid.
2.8 Effect of Atractylodes macrocephala lactone III on the anti-tumor effect of pyrroltinib
To investigate the medication safety of atractylenolide iii, the CompuSyn software was used to obtain a metering-reaction curve of atractylenolide iii and pyrroltinib and to obtain the CI value for the actual experimental point, which was found to be mostly below the equivalent line and mostly below 1.1, see (a) - (B) in fig. 12 and table 4. This result suggests that atractylenolide iii does not affect the anti-tumor effect of pyrroltinib.
TABLE 4 combination index CI values for actual experimental points
In a word, the bighead atractylodes rhizome lactone III can relieve diarrhea caused by the pyrroltinib, and the anti-tumor effect of the pyrroltinib is not affected. Atractylodes macrocephala lactone III can inhibit chloride ion secretion caused by pyrroltinib, and a potential action mechanism of the atractylenolide III is probably to increase AMPK phosphorylation and reduce protein expression of CFTR. Meanwhile, the atractylenolide III can relieve diarrhea caused by the pyrroltinib by adjusting the structure of intestinal flora and increasing the content of the lithocholic acid.
Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. Application of atractylenolide III in preparing medicine for preventing and treating diarrhea caused by tyrosine kinase inhibitor is provided.
2. The use of claim 1, wherein the tyrosine kinase inhibitor comprises trastuzumab, rituximab, lapatinib, lenatinib, pyrroltinib, afatinib, gefitinib, and erlotinib.
3. The use according to claim 1, wherein the prevention and/or treatment of diarrhea caused by tyrosine kinase inhibitors, in particular of diarrhea caused by pyrroltinib, is not affecting the antitumor effect of pyrroltinib.
4. The use according to claim 3, wherein said prevention and treatment of diarrhea caused by tyrosine kinase inhibitors is manifested by:
(a) The damage of the mucous membrane of the small intestine caused by the pyrroltinib is reduced;
(b) Inhibiting chloride ion secretion caused by pyrroltinib;
(c) Regulating the intestinal flora disorder caused by pyrroltinib;
(d) Increasing the content of the beneficial metabolite lithocholic acid.
5. The use of claim 4, wherein (b) is specifically: bighead atractylodes lactone III promotes AMPK phosphorylation and/or inhibits CFTR protein expression.
6. The use of claim 4, wherein (c) is specifically: bighead lactone III increases the levels of the beneficial flora wart and Achroman and decreases the levels of the adverse flora Clostridium to alleviate the diarrhea caused by pyrroltinib.
7. Application of tyrosine kinase inhibitor combined with atractylenolide III in preparing antitumor drug is provided.
8. The use according to claim 7, wherein the antitumor drug is embodied as: plays an anti-tumor effect of the tyrosine kinase inhibitor and prevents and/or treats diarrhea side effects mediated by the tyrosine kinase inhibitor;
The mass ratio of the tyrosine kinase inhibitor to the atractylenolide III is 16:1-3; or the mol ratio of the tyrosine kinase inhibitor to the atractylenolide III is 1-4:1000-25000.
9. The use of claim 8, wherein the tyrosine kinase inhibitor comprises trastuzumab, rituximab, lapatinib, lenatinib, pyrroltinib, afatinib, gefitinib, and erlotinib.
10. An antitumor drug, characterized in that the active ingredients thereof comprise a tyrosine kinase inhibitor and atractylenolide III;
Wherein the mass ratio of the tyrosine kinase inhibitor to the atractylenolide III is 16:1-3; or the mol ratio of the tyrosine kinase inhibitor to the atractylenolide III is 1-4:1000-25000;
The tyrosine kinase inhibitors include trastuzumab, rituximab, lapatinib, lenatinib, pyrroltinib, afatinib, gefitinib, and erlotinib.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410649279.XA CN118236369B (en) | 2024-05-24 | 2024-05-24 | Application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410649279.XA CN118236369B (en) | 2024-05-24 | 2024-05-24 | Application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN118236369A true CN118236369A (en) | 2024-06-25 |
| CN118236369B CN118236369B (en) | 2024-08-23 |
Family
ID=91556749
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410649279.XA Active CN118236369B (en) | 2024-05-24 | 2024-05-24 | Application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN118236369B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110548025A (en) * | 2019-09-11 | 2019-12-10 | 中国药科大学 | Application of atractylenolide II in preparation of medicine for improving insulin resistance and glycolipid metabolic disorder caused by obesity |
| CN113301921A (en) * | 2019-01-16 | 2021-08-24 | 加利福尼亚大学董事会 | Method for treating tyrosine kinase inhibitor-induced diarrhea |
-
2024
- 2024-05-24 CN CN202410649279.XA patent/CN118236369B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113301921A (en) * | 2019-01-16 | 2021-08-24 | 加利福尼亚大学董事会 | Method for treating tyrosine kinase inhibitor-induced diarrhea |
| CN110548025A (en) * | 2019-09-11 | 2019-12-10 | 中国药科大学 | Application of atractylenolide II in preparation of medicine for improving insulin resistance and glycolipid metabolic disorder caused by obesity |
Non-Patent Citations (2)
| Title |
|---|
| LAI ET AL.: "Effects of Shenling Baizhu powder on pyrotinib‑induced diarrhea: analysis of gut microbiota, metabonomics, and network pharmacology", CHINESE MEDICINE, vol. 140, no. 17, 31 December 2022 (2022-12-31), pages 1 - 16 * |
| XIAO ET AL.: "Repression of EZH2 Gene Expression by the Combination of Atractylenolide 1 and Erlotinib", CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, vol. 49, 14 September 2018 (2018-09-14), pages 1615 - 1632 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN118236369B (en) | 2024-08-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sun et al. | Resveratrol suppresses the growth and metastatic potential of cervical cancer by inhibiting STAT3Tyr705 phosphorylation | |
| JP2010508277A (en) | Methods for detecting and suppressing cancer | |
| Kim et al. | Targeting stem cells and dysplastic features with dual MEK/ERK and STAT3 suppression in gastric carcinogenesis | |
| US20170342503A1 (en) | Xrn2 as a determinant of sensitivity to dna damage | |
| WO2019237688A1 (en) | Application of niemann-pick c1 protein in diagnosis and treatment of cancer | |
| US20110224205A1 (en) | Use of curcumin or its analogues in cancer therapy utilizing epidermal growth factor receptor tyrosine kinase inhibitor | |
| WO2016148969A1 (en) | Kub5/hera as a determinant of sensitivity to dna damage | |
| CN118236369B (en) | Application of atractylenolide III in preventing and treating diarrhea caused by tyrosine kinase inhibitor | |
| CN111494351B (en) | Application of basic fuchsin in antitumor and medicine | |
| CN115960018B (en) | EGFR inhibitor, composition and application thereof | |
| EP3167887B1 (en) | Aryl amine substituted quinoxaline used as anticancer drugs | |
| CN111733235A (en) | Application of KDM5A gene and ATRX gene | |
| CN116271048A (en) | Application of SRC protein or down regulator of coding gene thereof in preparation of medicine for treating triple negative breast cancer | |
| CN111686111B (en) | Application of MALT1 protease inhibitor in preparation of non-small cell lung cancer therapeutic drug | |
| US20230172879A1 (en) | Therapeutic methods for preventing tumor metastasis and tumor recurrence | |
| Li et al. | Bushen Wenyang Huayu Decoction Targets TLR4/NF‐κB Mediated Autophagy to Treat Endometriosis Effectively | |
| Song et al. | Downregulation of miR‑7 and miR‑153 is involved in Helicobacter pylori CagA induced gastric carcinogenesis and progression | |
| CN109718374B (en) | Use of IRF3 inhibitor for preparing medicine for treating or preventing YAP over-activated cancer | |
| Pan et al. | Gancao Nurish-Yin Decoction medicated serum inhibits growth and migration of ovarian cancer cells: Network pharmacology-based analysis and biological validation | |
| JP2019501959A (en) | Use of Akt2 in tumor diagnosis and treatment | |
| CN110777204B (en) | Application of MAFG-AS1 knock-out reagent in preparation of medicines for treating bladder cancer | |
| CN107510843B (en) | DEM1 function and use | |
| AU2016319127A1 (en) | Medicinal Ambrosia plant extracts | |
| Li et al. | PNSC5325 prevents acute respiratory distress syndrome by alleviating inflammation and inhibiting extracellular matrix degradation of alveolar macrophages | |
| CN116694758A (en) | Application of LSD1 gene and related products |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |