CN115337300A - Use of water-soluble prodrugs of acacetin for the effective treatment of pulmonary hypertension - Google Patents
Use of water-soluble prodrugs of acacetin for the effective treatment of pulmonary hypertension Download PDFInfo
- Publication number
- CN115337300A CN115337300A CN202211060982.4A CN202211060982A CN115337300A CN 115337300 A CN115337300 A CN 115337300A CN 202211060982 A CN202211060982 A CN 202211060982A CN 115337300 A CN115337300 A CN 115337300A
- Authority
- CN
- China
- Prior art keywords
- acacetin
- water
- pulmonary hypertension
- pulmonary
- soluble prodrug
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- DANYIYRPLHHOCZ-UHFFFAOYSA-N 5,7-dihydroxy-4'-methoxyflavone Chemical compound C1=CC(OC)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 DANYIYRPLHHOCZ-UHFFFAOYSA-N 0.000 title claims abstract description 162
- 235000009962 acacetin Nutrition 0.000 title claims abstract description 81
- 229940002612 prodrug Drugs 0.000 title claims abstract description 79
- 239000000651 prodrug Substances 0.000 title claims abstract description 79
- 208000002815 pulmonary hypertension Diseases 0.000 title claims abstract description 72
- 238000011282 treatment Methods 0.000 title claims description 13
- 206010021143 Hypoxia Diseases 0.000 claims abstract description 52
- 108010041191 Sirtuin 1 Proteins 0.000 claims abstract description 52
- 102000000344 Sirtuin 1 Human genes 0.000 claims abstract description 52
- 102000055207 HMGB1 Human genes 0.000 claims abstract description 48
- 108700010013 HMGB1 Proteins 0.000 claims abstract description 48
- 101100339431 Arabidopsis thaliana HMGB2 gene Proteins 0.000 claims abstract description 46
- 101150021904 HMGB1 gene Proteins 0.000 claims abstract description 46
- 230000000694 effects Effects 0.000 claims abstract description 43
- 230000007954 hypoxia Effects 0.000 claims abstract description 33
- 210000001147 pulmonary artery Anatomy 0.000 claims abstract description 19
- 239000003814 drug Substances 0.000 claims abstract description 15
- 206010061218 Inflammation Diseases 0.000 claims abstract description 13
- 230000003111 delayed effect Effects 0.000 claims abstract description 13
- 230000004054 inflammatory process Effects 0.000 claims abstract description 13
- 230000002861 ventricular Effects 0.000 claims abstract description 12
- 230000004913 activation Effects 0.000 claims abstract description 10
- 208000018875 hypoxemia Diseases 0.000 claims abstract description 7
- 208000023504 respiratory system disease Diseases 0.000 claims abstract description 7
- 230000002265 prevention Effects 0.000 claims abstract description 3
- 210000004072 lung Anatomy 0.000 claims description 36
- 230000002685 pulmonary effect Effects 0.000 claims description 31
- 210000002565 arteriole Anatomy 0.000 claims description 23
- 208000000924 Right ventricular hypertrophy Diseases 0.000 claims description 9
- 230000002757 inflammatory effect Effects 0.000 claims description 8
- 238000007634 remodeling Methods 0.000 claims description 8
- 230000008719 thickening Effects 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 7
- 238000010254 subcutaneous injection Methods 0.000 claims description 7
- 239000007929 subcutaneous injection Substances 0.000 claims description 7
- 230000036772 blood pressure Effects 0.000 claims description 6
- 208000024891 symptom Diseases 0.000 claims description 6
- 230000009467 reduction Effects 0.000 claims description 5
- 102000011990 Sirtuin Human genes 0.000 claims description 3
- 108050002485 Sirtuin Proteins 0.000 claims description 3
- 230000001839 systemic circulation Effects 0.000 claims description 3
- 239000000556 agonist Substances 0.000 claims description 2
- 238000010253 intravenous injection Methods 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 230000003416 augmentation Effects 0.000 claims 1
- QVCMHGGNRFRMAD-XFGHUUIASA-N monocrotaline Chemical compound C1OC(=O)[C@](C)(O)[C@@](O)(C)[C@@H](C)C(=O)O[C@@H]2CCN3[C@@H]2C1=CC3 QVCMHGGNRFRMAD-XFGHUUIASA-N 0.000 abstract description 43
- QPNKYNYIKKVVQB-UHFFFAOYSA-N crotaleschenine Natural products O1C(=O)C(C)C(C)C(C)(O)C(=O)OCC2=CCN3C2C1CC3 QPNKYNYIKKVVQB-UHFFFAOYSA-N 0.000 abstract description 42
- QVCMHGGNRFRMAD-UHFFFAOYSA-N monocrotaline Natural products C1OC(=O)C(C)(O)C(O)(C)C(C)C(=O)OC2CCN3C2C1=CC3 QVCMHGGNRFRMAD-UHFFFAOYSA-N 0.000 abstract description 42
- 230000002829 reductive effect Effects 0.000 abstract description 13
- 201000010099 disease Diseases 0.000 abstract description 10
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 10
- 230000021736 acetylation Effects 0.000 abstract description 9
- 238000006640 acetylation reaction Methods 0.000 abstract description 9
- 230000001105 regulatory effect Effects 0.000 abstract description 8
- 208000006029 Cardiomegaly Diseases 0.000 abstract description 7
- 230000001684 chronic effect Effects 0.000 abstract description 7
- 238000001727 in vivo Methods 0.000 abstract description 7
- 230000002401 inhibitory effect Effects 0.000 abstract description 7
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 abstract description 6
- 208000029523 Interstitial Lung disease Diseases 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 6
- 201000002859 sleep apnea Diseases 0.000 abstract description 6
- 208000037129 Newborn Diseases Infant Diseases 0.000 abstract description 5
- 230000008569 process Effects 0.000 abstract description 3
- 230000001939 inductive effect Effects 0.000 abstract 1
- 238000011269 treatment regimen Methods 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 58
- 210000001519 tissue Anatomy 0.000 description 27
- 241000699666 Mus <mouse, genus> Species 0.000 description 16
- 230000007246 mechanism Effects 0.000 description 13
- 230000001146 hypoxic effect Effects 0.000 description 12
- 230000008859 change Effects 0.000 description 11
- 238000001514 detection method Methods 0.000 description 11
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 10
- 210000004204 blood vessel Anatomy 0.000 description 10
- 210000005241 right ventricle Anatomy 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 8
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 239000001301 oxygen Substances 0.000 description 8
- 238000011002 quantification Methods 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 238000010172 mouse model Methods 0.000 description 7
- 230000001225 therapeutic effect Effects 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 238000010186 staining Methods 0.000 description 6
- 230000035488 systolic blood pressure Effects 0.000 description 6
- 238000001262 western blot Methods 0.000 description 6
- 102000000849 HMGB Proteins Human genes 0.000 description 5
- 108010001860 HMGB Proteins Proteins 0.000 description 5
- 230000037396 body weight Effects 0.000 description 5
- 239000001569 carbon dioxide Substances 0.000 description 5
- 229910002092 carbon dioxide Inorganic materials 0.000 description 5
- 210000000805 cytoplasm Anatomy 0.000 description 5
- 230000003993 interaction Effects 0.000 description 5
- KMZHZAAOEWVPSE-UHFFFAOYSA-N 2,3-dihydroxypropyl acetate Chemical compound CC(=O)OCC(O)CO KMZHZAAOEWVPSE-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000000004 hemodynamic effect Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000012188 paraffin wax Substances 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 206010064911 Pulmonary arterial hypertension Diseases 0.000 description 3
- 208000032594 Vascular Remodeling Diseases 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000004872 arterial blood pressure Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 210000001715 carotid artery Anatomy 0.000 description 3
- 230000001086 cytosolic effect Effects 0.000 description 3
- 210000002889 endothelial cell Anatomy 0.000 description 3
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 3
- 230000028709 inflammatory response Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000001114 myogenic effect Effects 0.000 description 3
- 230000007959 normoxia Effects 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 238000004445 quantitative analysis Methods 0.000 description 3
- 230000003362 replicative effect Effects 0.000 description 3
- 230000009885 systemic effect Effects 0.000 description 3
- 230000035899 viability Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 208000014085 Chronic respiratory disease Diseases 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 206010020880 Hypertrophy Diseases 0.000 description 2
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 2
- 208000031481 Pathologic Constriction Diseases 0.000 description 2
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- QNVSXXGDAPORNA-UHFFFAOYSA-N Resveratrol Natural products OC1=CC=CC(C=CC=2C=C(O)C(O)=CC=2)=C1 QNVSXXGDAPORNA-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- AIGAZQPHXLWMOJ-UHFFFAOYSA-N Tanshinone I Chemical compound C1=CC2=C(C)C=CC=C2C(C(=O)C2=O)=C1C1=C2C(C)=CO1 AIGAZQPHXLWMOJ-UHFFFAOYSA-N 0.000 description 2
- LUKBXSAWLPMMSZ-OWOJBTEDSA-N Trans-resveratrol Chemical compound C1=CC(O)=CC=C1\C=C\C1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-OWOJBTEDSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 210000001168 carotid artery common Anatomy 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000000749 co-immunoprecipitation Methods 0.000 description 2
- 230000008602 contraction Effects 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- -1 flavonoid compound Chemical class 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 210000004731 jugular vein Anatomy 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 230000006911 nucleation Effects 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000011618 pulmonary hypertension animal model Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 235000021283 resveratrol Nutrition 0.000 description 2
- 229940016667 resveratrol Drugs 0.000 description 2
- 230000022379 skeletal muscle tissue development Effects 0.000 description 2
- 210000002460 smooth muscle Anatomy 0.000 description 2
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 230000036262 stenosis Effects 0.000 description 2
- 208000037804 stenosis Diseases 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 244000020998 Acacia farnesiana Species 0.000 description 1
- 235000003074 Acacia farnesiana Nutrition 0.000 description 1
- 244000061520 Angelica archangelica Species 0.000 description 1
- 206010003162 Arterial injury Diseases 0.000 description 1
- 206010003658 Atrial Fibrillation Diseases 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 229940127291 Calcium channel antagonist Drugs 0.000 description 1
- 206010007572 Cardiac hypertrophy Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 240000001879 Digitalis lutea Species 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 229940118365 Endothelin receptor antagonist Drugs 0.000 description 1
- 235000001287 Guettarda speciosa Nutrition 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 102100037907 High mobility group protein B1 Human genes 0.000 description 1
- 101710168537 High mobility group protein B1 Proteins 0.000 description 1
- 241000234435 Lilium Species 0.000 description 1
- 208000020165 Neonatal respiratory disease Diseases 0.000 description 1
- 229930189092 Notoginsenoside Natural products 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 206010037423 Pulmonary oedema Diseases 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- 206010039163 Right ventricular failure Diseases 0.000 description 1
- 241001047198 Scomberomorus semifasciatus Species 0.000 description 1
- 229930183118 Tanshinone Natural products 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 210000001008 atrial appendage Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 235000012467 brownies Nutrition 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000000480 calcium channel blocker Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000025084 cell cycle arrest Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 230000001877 deodorizing effect Effects 0.000 description 1
- 239000002274 desiccant Substances 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 210000004177 elastic tissue Anatomy 0.000 description 1
- 239000002308 endothelin receptor antagonist Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 210000002837 heart atrium Anatomy 0.000 description 1
- 230000004217 heart function Effects 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000002390 hyperplastic effect Effects 0.000 description 1
- 230000001969 hypertrophic effect Effects 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000003125 immunofluorescent labeling Methods 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000005550 inflammation mediator Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 210000005240 left ventricle Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004769 mitochondrial stress Effects 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000003387 muscular Effects 0.000 description 1
- 208000031225 myocardial ischemia Diseases 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229960003753 nitric oxide Drugs 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002590 phosphodiesterase V inhibitor Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 230000001110 prostacyclinlike Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000004088 pulmonary circulation Effects 0.000 description 1
- 230000009325 pulmonary function Effects 0.000 description 1
- 238000002407 reforming Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 208000013220 shortness of breath Diseases 0.000 description 1
- HUAUNKAZQWMVFY-UHFFFAOYSA-M sodium;oxocalcium;hydroxide Chemical compound [OH-].[Na+].[Ca]=O HUAUNKAZQWMVFY-UHFFFAOYSA-M 0.000 description 1
- 210000001562 sternum Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004026 tunica intima Anatomy 0.000 description 1
- 210000004231 tunica media Anatomy 0.000 description 1
- 229940124549 vasodilator Drugs 0.000 description 1
- 239000003071 vasodilator agent Substances 0.000 description 1
- 210000000596 ventricular septum 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
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/12—Antihypertensives
Abstract
The invention discloses an application of a water-soluble acacetin prodrug in effectively treating pulmonary hypertension. In the process of inducing pulmonary hypertension by using monocrotaline and hypoxia, the farnesoid water-soluble prodrug is found to be capable of reducing right ventricular pressure and right heart hypertrophy indexes and inhibiting pulmonary artery reconstruction; after the acacetin water-soluble prodrug is converted into acacetin in vivo, the activity and expression of SIRT1 can be up-regulated, acetylation activation and nuclear release of HMGB1 are inhibited, and subsequent delayed inflammatory reaction is reduced. The result shows that the acacetin and the acacetin water-soluble prodrug have the potential of being developed into a medicament for clinically preventing and treating pulmonary hypertension, and simultaneously provide a prevention and treatment strategy for respiratory diseases related to hypoxemia and/or inflammation, such as interstitial lung diseases, chronic obstructive lung diseases, sleep apnea syndromes, chronic altitude diseases, neonatal diseases and the like.
Description
Technical Field
The invention belongs to the field of medicines, and relates to application of a farnesoid or a farnesoid water-soluble prodrug in treating related diseases such as pulmonary hypertension.
Background
Pulmonary Hypertension (PH) is a disease caused by a variety of pathogenic factors, and is a disease in which the Pulmonary intra-arterial pressure is progressively increased, which in turn leads to increased Pulmonary circulation resistance and gradually leads to right ventricular hypertrophy, and right heart failure is induced. PH is not obvious in symptoms at the early stage of onset and lacks specificity, and patients only show symptoms of fatigue, shortness of breath, and the like, but progress rapidly, and once advanced, pulmonary artery remodeling (PVR) occurs, the mortality rate is extremely high, and the prognosis is very poor. Changes in the pathology characteristic of PVR mainly include: endothelial cells of the vascular intima are damaged, and swelling and hypertrophy appear; the smooth muscle cells of the tunica media of the blood vessels are significantly hyperplastic and hypertrophic; collagen fibers and elastic fibers of the adventitia are increased; some small blood vessels have thrombosis, non-muscular arteries form new muscle layers, and peripheral small blood vessels are myonized. Meanwhile, the pathological changes such as the enlargement of the main pulmonary artery and its branches, the hypertrophy of the right ventricle and the like can be seen.
The current clinical treatment methods for PH comprise taking anticoagulants, oxygen inhalation, diuresis, digitalis medicines, vasodilators, calcium channel blockers and the like. Other treatment approaches that have emerged in recent years have involved the use of phosphodiesterase type 5 inhibitors, prostacyclin-like drugs, nitric oxide, endothelin receptor antagonists, and the like. However, these treatments only improve the symptoms of patients and do not slow down the progress of PVR, so that there is an urgent need to search for a treatment capable of improving the cardiac and pulmonary functions of patients to improve the quality of life and prolong the life of patients. Studies show that the sirtuin 1 (SIRT 1) and the high mobility group protein B1 (HMGB 1) in the lung participate in the PVR process and promote the generation and development of PH. Wherein SIRT1 is important deacetylase in vivo and can inhibit acetylation activation of various inflammation mediators; HMGB1 is an important delayed inflammatory mediator in the lung, and can be activated by acetylation, so that nuclear release occurs and enters cytoplasm to start a downstream delayed inflammatory response and promote proliferation of cells. Therefore, regulatory measures against SIRT1 and HMGB1 are new entry points to reverse PVR and control PH. For example, SIRT1 activator "resveratrol" is used to increase the expression of SIRT1 in lung tissue, thereby inhibiting the development of pulmonary hypertension (see the research on the role and mechanism of SIRT1 in pulmonary hypertension vascular remodeling. Chinese academy of medicine science, 2010. Doi.
With the increasing emphasis on the role of traditional Chinese medicines in preventing and treating PH, many traditional Chinese medicines and extracts thereof are used for treating PH, including tanshinone, resveratrol, angelica, notoginsenoside, etc. However, these blood-activating stasis-resolving and blood-pressure-lowering medicinal components can improve the symptoms of pulmonary hypertension, such as right ventricular hypertrophy, and cause side effects such as lowering of systemic circulation blood pressure.
Acacia farnesiana is a flavonoid compound widely existing in various natural plants, has antioxidant, anti-inflammatory and anti-proliferative effects, and has protective effects on myocardial ischemia-reperfusion injury, atrial fibrillation, blood lipid reduction and the like. And the research shows that the water-soluble prodrug formed after the farnesoid is reformed can be directly converted into the farnesoid in beagle dogs, mice and rats. Thus, the absorption and utilization rate of the farnesoid in the body can be effectively improved without destroying the pharmacological action and generating other toxic substances (see Water-soluble acetin precursor conjugates biochemical analysis/diffusion in scientific reports. DOI:10.1038/srep36435. And Synthesis of a high-soluble Water-soluble acetin precursor for extracting effective catalytic reaction in biological reports. DOI:10.1038/srep 25743.). However, there are no reports of the therapeutic action and mechanism of PH treatment of acacetin and its water-soluble prodrug.
The component of the composition for protecting cells from the influence of oxidation and mitochondrial stress disclosed in Chinese patent CN112423726A relates to acacetin, and the disease treated by the composition relates to skin conditions related to inflammatory skin diseases. However, the patent does not disclose that the composition can affect the SIRT1-HMGB1 pathway and the downstream delayed inflammatory response regulated by the SIRT1-HMGB1 pathway.
Disclosure of Invention
The purpose of the present invention is to provide the use of a water-soluble prodrug of farnesoid for effectively treating pulmonary hypertension.
In order to achieve the purpose, the invention adopts the following technical scheme:
application of acacetin or acacetin water-soluble prodrug in preparing medicines for preventing and treating pulmonary hypertension.
Preferably, the pulmonary hypertension is caused by inflammation and/or hypoxia of lung tissues, and the animal model for evaluating the curative effect is formed by the induction of monocrotaline and hypoxia.
Preferably, the acacetin water-soluble prodrug is administered to pulmonary hypertension model mice (dose of 20 mg/kg) by subcutaneous injection once a day for 2 weeks.
Preferably, the acacetin or acacetin water-soluble prodrug is used for treating one or more of the following symptoms of induced pulmonary hypertension: increased right ventricular pressure (e.g., right ventricular systolic pressure), right ventricular hypertrophy (e.g., right ventricular hypertrophy), pulmonary artery remodeling, pulmonary arteriole wall thickening and luminal narrowing, and myelinated pulmonary arteriole.
Preferably, the acacetin or acacetin water-soluble prodrug does not cause a decrease in systemic blood pressure while treating induced pulmonary hypertension.
Preferably, the acacetin inhibits activation and release of late inflammatory mediators by up-regulating the activity and expression of pulmonary SIRT 1.
Preferably, the delayed inflammatory mediator is HMGB1.
Preferably, the acacetin water-soluble prodrug is formed by reforming a fat-soluble molecule of acacetin, and the acacetin water-soluble prodrug is converted into acacetin in vivo, so that the acacetin water-soluble prodrug is suitable for being developed into an injection preparation or an oral preparation for clinical use.
Application of acacetin or acacetin water-soluble prodrug in preparation of medicine for preventing and treating pulmonary hypertension complications is provided.
Use of acacetin or a water-soluble prodrug of acacetin for the manufacture of a medicament for the prevention or treatment of a respiratory disease associated with hypoxemia and/or inflammation.
Preferably, the respiratory disease is selected from clinically common interstitial lung diseases, chronic obstructive lung diseases, sleep apnea syndrome, chronic altitude diseases or some neonatal respiratory diseases.
Preferably, the medicine is used for subcutaneous injection, intravenous injection or oral administration, and the water-soluble acacetin prodrug is converted into acacetin in vivo, so that the absorption and utilization of the acacetin are improved.
Application of acacetin in preparing SIRT1 agonist is provided.
The invention has the beneficial effects that:
the invention establishes a pulmonary hypertension model, gives a water-soluble prodrug of the model acacetin and observes the curative effect. The result shows that the acacetin water-soluble prodrug can effectively reduce right ventricular pressure and right heart hypertrophy index after being converted into acacetin in vivo, and inhibit pulmonary artery remodeling, thereby achieving the effect of treating pulmonary hypertension. And the acacetin water-soluble prodrug can not cause the change of the hemodynamics and the morphology of the pulmonary arteriole of the normal control, can not improve the pulmonary artery reconstruction and treat the pulmonary hypertension, and can not generate the side effect of causing the reduction of the systemic circulation blood pressure. The invention not only provides a new strategy for preventing and treating pulmonary hypertension, but also is suitable for preventing and treating respiratory system diseases related to hypoxemia and/or inflammation, such as interstitial lung diseases, chronic obstructive pulmonary diseases, sleep apnea syndrome, chronic altitude diseases, neonatal diseases and the like, and has good application prospect.
Furthermore, the invention defines the key mechanism of acacetin for treating pulmonary hypertension: the activity and the expression of lung SIRT1 are up-regulated, so that acetylation activation and nuclear release of a delayed inflammatory medium HMGB1 are inhibited, subsequent delayed inflammatory reaction is reduced, and the treatment effects of inhibiting pulmonary artery reconstruction, reducing right ventricular pressure and improving right ventricular hypertrophy are achieved.
Drawings
FIG. 1 shows the chemical structure of water-soluble prodrug of farnesoid (A) and the scheme (B) of experiment for interfering with the mouse pulmonary hypertension induced by Monocrotaline (MCT) and hypoxia (hypoxia) and water-soluble prodrug of farnesoid (Acacetin).
FIG. 2 is a graph showing the experimental results of the effectiveness of treatment of Monocrotaline (MCT) -induced pulmonary hypertension in mice; a: body weight (Body weight) changes in the differently treated mice over 6 weeks (./P <0.05, n = 10); b: change in the effect of administration of a water-soluble prodrug of farnesoid on the right ventricular systolic blood pressure peak (RVSP) in mice (./P <0.05, n = 10); c: changes in the effect of administration of a water-soluble prodrug of acacetin on mean carotid artery pressure (mCAP) in mice; d: change in the effect of administration of a water-soluble prodrug of acacetin on the right heart hypertrophy index (RV/(LV + S)%) (. P <0.05, n = 10) in mice; e: changes in the effect of administration of water-soluble prodrugs of farnesoid on lung histomorphology in mice (HE staining test); f: change in the effect of administration of a water-soluble prodrug of acacetin on the pulmonary arteriole area ratio (WA%) in mice (./P <0.05, n = 10); g: change in the effect of administration of a water-soluble prodrug of acacetin on pulmonary arteriole diameter ratio (WT%) in mice (./P <0.05, n = 10); h: changes in the effect of administration of water-soluble prodrugs of farnesoid on pulmonary arteriolar myogenesis (detection of α -SMA staining); i: quantitative analysis of pulmonary arteriole α -SMA staining (./P <0.05, n = 4).
FIG. 3 is a graph of experimental demonstration of the mechanism of therapeutic action of farnesoid (up-regulation of SIRT1, thereby inhibiting acetylation activation and nucleation of HMGB 1) in a Monocrotaline (MCT) -induced pulmonary hypertension model in mice; a: results of detection of SIRT1 viability in lung tissue of different treated mice (× P <0.05, n = 10); b: results of measurements of HMGB1 content in lung tissue of mice treated differently (× P <0.05, n = 10); c: changes in SIRT1, HMGB1 and ac-HMGB1 protein expression in lung tissue of differently treated mice (Western Blot assay); d: quantification of SIRT1 protein expression (× P <0.05, n = 3); e: quantification of HMGB1 protein expression (./P <0.05, n = 3); f: quantification of ac-HMGB1 protein expression (./P <0.05, n = 3); g: quantification result of ac-HMGB1/HMGB1 (× P <0.05, n = 3); h: the Co-ip detection result (the interaction between SIRT1 and HMGB 1); i: HMGB1 expression changes in the nucleus (Nuclear) and cytoplasm (Cytoplasmic).
FIG. 4 is a graph showing the experimental results of the effectiveness of treatment of hypoxia (hypoxia) -induced pulmonary hypertension in mice. A: body weight (Body weight) changes in the differently treated mice over 6 weeks (./P <0.05, n = 10); b: change in the effect of administering a water-soluble prodrug of farnesoid on right ventricular systolic blood pressure (RVSP) in mice (× P <0.05, n = 10); c: changes in the effect of administration of a water-soluble prodrug of acacetin on mean carotid artery pressure (mCAP) in mice; d: change in the effect of administration of a water-soluble prodrug of acacetin on the right heart hypertrophy index (RV/(LV + S)%) (. P <0.05, n = 10) in mice; e: change in the effect of administration of a water-soluble prodrug of acacetin on the pulmonary arteriole area ratio (WA%) in mice (./P <0.05, n = 10); f: change in the effect of administration of a water-soluble prodrug of acacetin on pulmonary arteriole diameter ratio (WT%) in mice (./P <0.05, n = 10); g: quantitative analysis of pulmonary arteriole α -SMA staining (× P <0.05, n = 4); h: changes in the effect of administration of water-soluble prodrugs of farnesoid on lung histomorphology in mice (HE staining test); i: changes in the effect of administration of water-soluble prodrugs of farnesoid on pulmonary arteriolar myogenesis (detection of α -SMA staining).
FIG. 5 is a graph of the results of experimental demonstration of the mechanism of therapeutic action of farnesoid (up-regulation of SIRT1, thereby inhibiting acetylation activation and nucleation of HMGB 1) in a hypoxia (hypoxia) -induced pulmonary arterial hypertension model in mice; a: results of detection of SIRT1 viability in lung tissue of differently treated mice (./P <0.05, n = 10); b: results of measurements of HMGB1 content in lung tissue of mice treated differently (./P <0.05, n = 10); c: changes in SIRT1, HMGB1 and ac-HMGB1 protein expression in lung tissue of differently treated mice (Western Blot assay); d: quantification of SIRT1 protein expression (× P <0.05, n = 3); e: quantification of ac-HMGB1 protein expression (./P <0.05, n = 3); f: quantification of HMGB1 protein expression (./P <0.05, n = 3); g: quantification of ac-HMGB1/HMGB1 (× P <0.05, n = 3); h: the Co-ip detection result (the interaction between SIRT1 and HMGB 1); i: HMGB1 expression changes in the nucleus (Nuclear) and cytoplasm (Cytoplasmic).
Detailed Description
The invention is described in further detail below with reference to the figures and examples. The following examples are only for illustrating the present invention and are not intended to limit the scope of the present invention.
According to the invention, the acacetin water-soluble prodrug is adopted to treat pulmonary hypertension, and the acacetin water-soluble prodrug can be converted into acacetin in vivo, so that the relative curative effect of the acacetin water-soluble prodrug after administration is evaluated by using monocrotaline and hypoxia-induced pulmonary hypertension model, and the action mechanism of the acacetin formed by in vivo conversion in treating pulmonary hypertension is clarified. The invention also provides experimental basis for preventing and treating respiratory system diseases related to hypoxemia and/or hypoxemia, such as interstitial lung diseases, chronic obstructive pulmonary diseases, sleep apnea syndrome, chronic altitude diseases, neonatal diseases and the like.
1. Liquid medicine preparation, pulmonary hypertension animal model construction and experimental grouping
Water-soluble farnesin Water-soluble farnesin prodrug, which is shown in FIG. 1A, can be converted directly into farnesin beagle dogs, mice and rats by techniques and means such as high performance liquid chromatography (Water-soluble acetin prodrug, amplification, scientific reports, DOI:10.1038/srep36435., and Synthesis of a high Water-soluble acetin prodrug for treating experimental reaction, DOI:10.1038/srep 25743.) in the literature. Reference is made to the dissolution of water-soluble prodrugs of farnesoids in sterile ddH 2 And O, preparing a concentration injection of 40mg/mL, and administering monocrotaline and hypoxia-induced pulmonary hypertension mice in a subcutaneous injection mode to observe the treatment effect and define an effect mechanism.
C57BL/6 mice (female and male halves, 7-8 weeks old, weight about 18-22 g) were randomly divided into 8 groups of 10 mice each, and the specific grouping was illustrated below with reference to FIG. 1B:
(1) control group (Control): mice were injected subcutaneously with 0.9% normal saline at one time.
(2) Acacetin water soluble prodrug Control group I (Control + Acacetin): mice were injected subcutaneously with 0.9% normal saline at one time. Farnesoid water-soluble prodrug (20 mg/kg) was administered by subcutaneous injection once daily for 2 weeks at week 4.
(3) Monocrotaline group: mice were injected subcutaneously with 60mg/kg Monocrotaline (MCT) in one portion, observed for 6 weeks, and replicated the mouse monocrotaline-induced pulmonary hypertension model.
(4) Acacetin water-soluble prodrug intervention group I (MCT + Acacetin): mice were injected subcutaneously once with 60mg/kg MCT and the water-soluble prodrug of farnesoid (20 mg/kg) was administered subcutaneously once daily for 2 weeks at week 4.
(5) Normoxia (Normoxia): mice were placed in an normoxic environment for 6 weeks.
(6) Acacetin water-soluble prodrug control group II (Normoxia + Acacetin): mice were placed in an normoxic environment and given a water-soluble prodrug of farnesoid (20 mg/kg) by subcutaneous injection once daily for 2 weeks at week 4.
(7) Hypoxic group (Hypoxia): mice were placed in a hypoxic chamber with oxygen concentration of about 10% and carbon dioxide concentration of about 5% for 8h each day for 6 weeks, replicating the mouse hypoxic pulmonary hypertension model.
(8) Acacetin water-soluble prodrug intervention group II (Hypoxia + Acacetin): mice were placed daily in a hypoxic chamber under reduced pressure of about 10% oxygen and about 5% carbon dioxide for 8h, and farnesoid water-soluble prodrug (20 mg/kg) was administered by subcutaneous injection once daily for 2 weeks at week 4.
Mice in normoxic groups were housed in animal chambers and raised naturally (atmospheric pressure approximately 718mm Hg 2 150.6mmHg, oxygen concentration about 21%); hypoxic mice were placed in a hypoxic chamber at low pressure (chamber pressure of 380mm hg 2 Reduced to 79.6mmHg, corresponding to an oxygen content of 5540 meters above sea level, with an oxygen concentration of about 10% and a carbon dioxide concentration of about 5%) for 6 weeks per day; deodorizing and absorbing CO with soda lime and desiccant in low-pressure and low-oxygen chamber 2 . All mice were treated and examined for pulmonary hypertension-related indications at week 6 to evaluate the therapeutic effect of farnesoid water-soluble prodrug on pulmonary hypertension mice. Meanwhile, in order to clarify the action mechanism of farnesoid for treating MCT and hypoxia-induced pulmonary hypertension, the activity, content and expression changes of SIRT1, HMGB1 and ac-HMGB1 in lung tissues are detected.
2. Hemodynamic index detection
Anaesthetizing and fixing the mouse, making an incision in the center of the neck, separating and ligating the far-center ends of the left common carotid artery and the right external jugular vein, clamping the near-center end of the blood vessel, cutting a small opening between the two by an ophthalmic scissors, respectively inserting a polyethylene catheter filled with 0.5% heparin solution into the left common carotid artery and the right external jugular vein, leaving one end of the catheter in the blood vessel, knotting and fixing, connecting the other end of the catheter with a pressure transducer, and recording the mean carotid artery pressure (mCAP) and the right ventricular systolic pressure peak value (RVSP) of the mouse.
3. Measurement of Right Heart hypertrophy index (RV/(LV + S)%)
The sternum of the mouse is cut open, and the heart is exposed; removing tissue and blood vessels around the heart, left and right atria, atrial appendage, etc., finding the pulmonary artery cone, cutting the Right Ventricle (RV) down the pulmonary artery cone and weighing; the remaining tissue was also weighed, i.e., the weight of the left ventricle and the ventricular septum (LV + S); calculating the right heart hypertrophy index: RV/(LV + S). Times.100% to reflect the degree of right ventricular hypertrophy.
4. Lung tissue paraffin section preparation and HE staining
The material is taken along the transverse section of the pulmonary portal, a tissue block of about 1cm multiplied by 2cm on the upper lobe of the right lung of the mouse is cut and placed in an embedding frame, the embedding frame and the embedding frame are placed in 10 percent neutral formaldehyde buffer solution for fixation for 24 hours, and then the embedding frame is taken out and placed in 70 percent ethanol solution. Then dehydrated, embedded, and made into paraffin blocks. Paraffin blocks were sectioned, deparaffinized to water and HE stained to detect changes in pulmonary arterioles.
5. Quantitative analysis of pulmonary arterioles
Observing the HE stained section under a microscope, selecting small pulmonary arteries with the outer diameter of less than 50-100 mu m, collecting and analyzing blood vessel images by using image analysis software, respectively measuring the inner diameter, the outer diameter, the wall thickness and the blood vessel area of the blood vessel, and respectively calculating two indexes reflecting the thickening of the blood vessel wall, namely WT% (the wall thickness/the outer diameter multiplied by 100%) and WA% (the wall area/the total area multiplied by 100%) according to the measured values.
6. Immunofluorescence staining
Paraffin sections of mouse lung tissue were dewaxed to water and infiltrated with 0.1% Triton-100, 1% BSA for 1 hour. alpha-SMA primary antibody was added separately and incubated overnight at 4 ℃. After washing 3 times with PBS, a fluorescent secondary antibody (594 nm) was added and incubated at room temperature for 1 hour. And finally, adding DAPI to incubate for 5 minutes, and scanning and observing the expression condition of the alpha-SMA in the lung tissue of the mouse by using a laser confocal microscope.
7.SIRT1 Activity detection
Determining the activity of the mouse lung tissue SIRT1 by using a SIRT1 activity detection kit: SIRT1 protein in lung tissue lysates was collected by immunoprecipitation, eluted, quantitated and diluted with gradient buffer in the kit. The substrate and reagent were added to a 96-well plate and reacted at room temperature for 30 minutes. The viability of SIRT1 was then quantified using a spectrophotometer.
8. detection of HMGB1 content by ELISA method
The right lower lung lobe tissue was taken, 100mg of the tissue was accurately weighed and placed in an EP tube, 1mL of physiological saline was added, and ground with a hand-held homogenizer to prepare a tissue homogenate. Then, the mixture was centrifuged at 4500rpm at 4 ℃ for 10min, and the supernatant was aspirated. Detecting the content of HMGB1 in lung tissues according to the operation of an ELISA kit specification.
9. Detecting expression changes of SIRT1, HMGB1 and ac-HMGB1 in mouse lung tissues
Western Blot: taking the left lower lung lobe tissue, accurately weighing 100mg of the tissue, and extracting and quantifying the lung tissue protein. Then, the expression changes of SIRT1, HMGB1 and ac-HMGB1 are analyzed through links such as vertical gel electrophoresis, membrane transfer, antibody combination, chemiluminescence, gel imaging and the like (the SIRT1 and HMGB1 antibodies are purchased from Abcam antibody company, the goods numbers are ab110304 and ab79823 respectively; the ac-HMGB1 antibody is purchased from Wuhan ABClonal company, the goods number is A16002).
Next, nuclear and cytoplasmic fractions in lung tissue were isolated, and the levels of HMGB1 in the nucleus and cytoplasm were examined to evaluate the activity of HMGB1 entering the nucleus for transcription, respectively.
Finally, co-immunoprecipitation (co-IP) was performed using the kit, and the interaction of SIRT1 with HMGB1 was analyzed: the SIRT1 antibody was immobilized on a coupling resin, and then lung tissue lysates were prewashed and incubated with the immobilized resin overnight at 4 ℃. And finally, eluting the protein and carrying out Western blot analysis and detection.
10. Results of the experiment
(1) Therapeutic effect on mouse pulmonary hypertension induced by monocrotaline
The mechanism of the Monocrotaline (MCT) is that the MCT is converted into lilium brownie pyrrole in the liver to cause cell cycle arrest, apoptosis and perivascular inflammation of pulmonary artery endothelial cells and promote vascular intimal stripping, thereby causing the progressive proliferation of pulmonary artery smooth muscle cells and pulmonary artery remodeling. In view of the similarity of the characteristics of MCT model with the physiological and pathological mechanisms of human pulmonary hypertension pathogenesis, MCT is often used to duplicate the pulmonary hypertension animal model so as to better understand the process of pulmonary artery remodeling and the important role of inflammatory reaction in the pathogenesis.
MCT-induced pulmonary hypertension mice (MCT mice for short) gradually showed anorexia, no seminal fluid withdrawal, and insignificant weight gain (Body weight) over the observation period of 6 weeks (fig. 2A); by comparing MCT pulmonary hypertension mouse models replicating for 6 weeks (i.e., MCT mice) with mice from the control group: the right ventricular systolic pressure peak (RVSP) of MCT mice WAs significantly increased (fig. 2B), which reflects that the indicator of right ventricular hypertrophy (i.e., RV/(LV + S)%) WAs also significantly increased (fig. 2D), lung histological evaluation showed significant alveolar edema, large-lamellar exudation and hemorrhage, significant thickening of pulmonary arteriole smooth muscle layer, luminal stenosis (fig. 2E), and significant increase in both WA% and WT% which reflect the indicators of pulmonary arteriole thickening (fig. 2F, fig. 2G). Pulmonary arteriole a-SMA staining results showed myogenic pulmonary arteriole increase in MCT mice (fig. 2H, fig. 2I), indicating that Monocrotaline (MCT) is able to induce significant pulmonary hypertension. The administration of acacetin water-soluble prodrug (structure shown in fig. 1A) was effective in improving the general condition of MCT mice and increasing the weight of mice (fig. 2B), and the manifestation of pulmonary hypertension was significantly reduced after the administration of acacetin water-soluble prodrug (fig. 2B to fig. 2I), indicating that the administration of acacetin water-soluble prodrug was effective in improving MCT-induced pulmonary hypertension. It is worth mentioning that the acacetin water-soluble prodrug did not cause hemodynamic and pulmonary arteriole morphological changes in normal mice (i.e., control mice), and did not cause a decrease in systemic blood pressure while ameliorating MCT-induced pulmonary arterial remodeling and pulmonary arterial hypertension (fig. 2C).
(2) Effect and interaction of SIRT1, HMGB1 in a mouse model of MCT pulmonary hypertension after administration of Water-soluble prodrugs of farnesoid
The SIRT1 activity and the Western Blot experiment result show that the SIRT1 activity in the lung of an MCT mouse is obviously reduced, and the HMGB1 content is obviously increased (fig. 3A and fig. 3B). And MCT resulted in a significant reduction in the expression level of SIRT1 (fig. 3C, fig. 3D), while the expression of total HMGB1 and ac-HMGB1 was significantly increased, with ac-HMGB1 being the major component of activated HMGB1 (fig. 3C, fig. 3E, fig. 3F and fig. 3G). The results of the Co-IP experiments showed that SIRT1 was able to bind directly to HMGB1, but SIRT1 binding to HMGB1 was significantly reduced by MCT (fig. 3H). Furthermore, MCT promoted the release of HMGB1 out of the nucleus, i.e. a significant increase of HMGB1 entering the cytoplasm (fig. 3I). By administering the acacetin water-soluble prodrug, the activity of SIRT1 in lungs of MCT mice can be increased, the expression of SIRT1 is up-regulated, the content of HMGB1 is reduced, the expression of total HMGB1 and ac-HMGB1 is down-regulated, the combination of SIRT1 and HMGB1 is promoted, and the nuclear release of HMGB1 is inhibited (fig. 3A-fig. 3I). The above results suggest that farnesin plays a role mechanism for treating MCT-induced pulmonary hypertension, i.e., farnesin (a water-soluble prodrug of farnesin is converted into farnesin vivo) increases the activity of SIRT1 in the lung and promotes the expression of SIRT1, thereby inhibiting acetylation activation and nuclear release of the delayed inflammatory mediator HMGB1 and reducing the subsequent delayed inflammatory response.
(3) Therapeutic effect on hypoxia-induced pulmonary hypertension of mice
Hypoxia is also one of the ways to induce pulmonary hypertension. Hypoxia causes injury to pulmonary artery endothelial cells, and an imbalance in the vascular factors involved in regulating pulmonary artery contraction/relaxation, increases the pulmonary artery contraction response and promotes arterial remodeling, ultimately leading to pulmonary hypertension. Pulmonary hypertension caused by chronic hypoxia is commonly seen in clinical various chronic respiratory diseases, such as interstitial lung diseases, chronic obstructive lung diseases, sleep apnea syndrome, chronic altitude diseases, certain neonatal diseases and the like. For example, the animal is placed in a low-pressure hypoxia chamber and is continuously hypoxic for 2-8 weeks (oxygen concentration in the chamber is maintained at about 10% and carbon dioxide concentration is maintained at about 5%), and at the moment, the model animal is easy to have blood gas change of hypoxia and high carbon dioxide, so that the model animal is more suitable for the real situation of clinical chronic respiratory disease patients.
As a result of comparison with a mouse model of hypoxic pulmonary hypertension replicating for 6 weeks (i.e., hypoxia-induced pulmonary hypertension mouse, abbreviated as hypoxic mouse), it was shown that administration of farnesoid water-soluble prodrug (fig. 1A) improves the general condition of hypoxic mice and significantly increases the weight of mice within the observation period of 6 weeks (fig. 4A). And also significantly inhibited hypoxia-induced increases in RVSP, RV/(LV + S)%, WA% and WT% after administration of the acacetin water-soluble prodrug (FIG. 4B, FIG. 4D, FIG. 4E and FIG. 4F). Lung histological evaluation showed that administration of the water-soluble prodrug of farnesoid reversed the thickening of the pulmonary arteriolar smooth muscle layer and luminal narrowing in mice caused by hypoxia (fig. 4H). Pulmonary arteriole a-SMA staining results show that administration of the water-soluble prodrug of farnesoid significantly reduced the myogenic pulmonary arterioles in hypoxic mice (fig. 4G, fig. 4I). The above results indicate that administration of a water-soluble prodrug of farnesoid also has significant efficacy against hypoxia-induced pulmonary hypertension. Furthermore, the water-soluble prodrug of acacetin did not cause changes in the hemodynamics and pulmonary arteriole morphology of normoxic mice (i.e., normoxic mice), and did not cause a decrease in systemic blood pressure while ameliorating hypoxia-induced pulmonary arterial remodeling and pulmonary arterial hypertension (fig. 4C).
(4) Effect and interaction of SIRT1, HMGB1 in a hypoxic pulmonary hypertension mouse model following administration of Water-soluble prodrugs of farnesoid
The SIRT1 activity and the Western Blot experiment result show that hypoxia also reduces the SIRT1 activity in the mouse lung and increases the HMGB1 content (fig. 5A and 5B). And hypoxia also resulted in a significant reduction in SIRT1 expression (fig. 5C, 5D) and increased ac-HMGB1 and total HMGB1 expression (fig. 5C, 5E, 5F and 5G). The results of the Co-IP experiments show that hypoxia inhibits the binding of SIRT1 and HMGB1 (FIG. 5H), and promotes the nuclear release of HMGB1 (FIG. 5I). By administering a water-soluble prodrug of acacetin, the expression change of SIRT1 and HMGB1 due to hypoxia can be improved, the binding of SIRT1 and HMGB1 can be promoted, and the nuclear release of HMGB1 can be inhibited (fig. 5A to 5I). The above results suggest a mechanism of action of farnesin treating hypoxia-induced pulmonary hypertension, that is, farnesin (a water-soluble prodrug of farnesin is converted into farnesin vivo) increases the activity of SIRT1 in the lung and promotes the expression of SIRT1, thereby inhibiting acetylation activation and nuclear release of a delayed inflammatory mediator HMGB1 and reducing subsequent delayed inflammatory reaction.
11. Conclusion of the experiment
Experiments respectively prove that the therapeutic effects of subcutaneously injecting the acacetin water-soluble prodrug on the monocrotaline and hypoxia-induced pulmonary hypertension are as follows: in an observation period of 6 weeks, general state difference and weight of MCT and hypoxia-induced pulmonary hypertension mouse models are effectively improved, right ventricular systolic pressure peak (RVSP) of the pulmonary hypertension mouse models and index RV/(LV + S)%, which reflects right ventricular hypertrophy, are reduced, reconstruction of pulmonary arteries is inhibited, indexes WA% and WT, which reflect pulmonary arteriolar thickening, lumen stenosis and wall thickening of the pulmonary arterioles are improved, and formation of myogenic pulmonary arterioles is reduced.
In conclusion, the invention discloses that the acacetin water-soluble prodrug can be used as an effective medicine component for treating pulmonary hypertension (the index of right ventricular pressure and right cardiac hypertrophy is reduced, pulmonary artery remodeling is improved), and a mechanism for taking effect of the acacetin through the influence on a SIRT1-HMGB1 pathway (the activity and the expression of SIRT1 in the lung are up-regulated, so that acetylation activation and nuclear release of HMGB1 are inhibited, and subsequent delayed inflammatory reaction is reduced) is defined. Based on the key mechanism for treating the pulmonary hypertension and the combination of the experimental results of relevant animal models, the invention not only provides a new strategy for preventing and treating the pulmonary hypertension, but also provides ideas and strategies for preventing and treating a series of respiratory diseases related to hypoxemia and/or inflammation, such as interstitial lung diseases, chronic obstructive lung diseases, sleep apnea syndrome, chronic altitude diseases, neonatal diseases and the like.
Claims (10)
1. Application of acacetin or acacetin water-soluble prodrug in preparation of medicine for preventing and treating pulmonary hypertension.
2. Use according to claim 1, characterized in that: the pulmonary hypertension is caused by inflammation and/or hypoxia of the lung tissue.
3. Use according to claim 1, characterized in that: the acacetin or acacetin water-soluble prodrug is used for treating one or more of the following symptoms of pulmonary hypertension: increased right ventricular pressure, right ventricular hypertrophy, pulmonary artery remodeling, pulmonary arteriole wall thickening and luminal narrowing, and myelinated pulmonary arteriole augmentation.
4. Use according to claim 1, characterized in that: the acacetin or acacetin water-soluble prodrug can treat pulmonary hypertension without causing reduction of systemic circulation blood pressure.
5. Use according to claim 1, characterized in that: the acacetin inhibits activation and release of late inflammatory mediators by up-regulating the activity and expression of pulmonary SIRT 1.
6. Use according to claim 5, characterized in that: the delayed inflammatory mediator is HMGB1.
7. Application of acacetin or acacetin water-soluble prodrug in preparation of medicine for preventing and treating pulmonary hypertension complications is provided.
8. Use of acacetin or a water-soluble prodrug of acacetin for the manufacture of a medicament for the prevention or treatment of a respiratory disease associated with hypoxemia and/or inflammation.
9. Use according to claim 1, 7 or 8, characterized in that: the medicine is used for subcutaneous injection, intravenous injection or oral administration.
10. Application of acacetin or acacetin water-soluble prodrug in preparing SIRT1 agonist.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211060982.4A CN115337300A (en) | 2022-08-31 | 2022-08-31 | Use of water-soluble prodrugs of acacetin for the effective treatment of pulmonary hypertension |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211060982.4A CN115337300A (en) | 2022-08-31 | 2022-08-31 | Use of water-soluble prodrugs of acacetin for the effective treatment of pulmonary hypertension |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115337300A true CN115337300A (en) | 2022-11-15 |
Family
ID=83955893
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211060982.4A Pending CN115337300A (en) | 2022-08-31 | 2022-08-31 | Use of water-soluble prodrugs of acacetin for the effective treatment of pulmonary hypertension |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115337300A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110171193A1 (en) * | 2008-06-12 | 2011-07-14 | The Board Of Trustees Of The University Of Illinois | Compositions and methods for treating pulmonary hypertension |
-
2022
- 2022-08-31 CN CN202211060982.4A patent/CN115337300A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110171193A1 (en) * | 2008-06-12 | 2011-07-14 | The Board Of Trustees Of The University Of Illinois | Compositions and methods for treating pulmonary hypertension |
Non-Patent Citations (2)
| Title |
|---|
| LI-CHAO SUN ET AL: "Protective effect of acacetin on sepsis-induced acute lung injury via its anti-inflammatory and antioxidative activity", 《ARCH. PHARM. RES.》, no. 41, pages 1199 - 1210 * |
| 王琳等: "基于网络药理学探讨桔梗治疗肺动脉高压的作用机制", 《湖南中医杂志》, vol. 38, no. 2, pages 149 - 155 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Ablation of endothelial Pfkfb3 protects mice from acute lung injury in LPS-induced endotoxemia | |
| Kume et al. | Pioglitazone attenuates inflammatory atrial fibrosis and vulnerability to atrial fibrillation induced by pressure overload in rats | |
| US11433088B2 (en) | Methods for treatment of vascular endothelial dysfunction using nicotinamide mononucleotide | |
| Yan et al. | Baicalin attenuates hypoxia-induced pulmonary arterial hypertension to improve hypoxic cor pulmonale by reducing the activity of the p38 MAPK signaling pathway and MMP-9 | |
| Zhang et al. | Electroacupuncture preconditioning attenuates acute myocardial ischemia injury through inhibiting NLRP3 inflammasome activation in mice | |
| Kang et al. | Sulforaphane prevents right ventricular injury and reduces pulmonary vascular remodeling in pulmonary arterial hypertension | |
| Hobolth et al. | Effects of carvedilol and propranolol on circulatory regulation and oxygenation in cirrhosis: a randomised study | |
| Andrade et al. | Ablation of brainstem C1 neurons improves cardiac function in volume overload heart failure | |
| Hua et al. | Metformin increases cardiac rupture after myocardial infarction via the AMPK-MTOR/PGC-1α signaling pathway in rats with acute myocardial infarction | |
| Chen et al. | Treatment of stress urinary incontinence by cinnamaldehyde, the major constituent of the chinese medicinal herb ramulus cinnamomi | |
| Liao et al. | PRDX6-mediated pulmonary artery endothelial cell ferroptosis contributes to monocrotaline-induced pulmonary hypertension | |
| Bai et al. | Continuous infusion of angiotensin IV protects against acute myocardial infarction via the inhibition of inflammation and autophagy | |
| Zhang et al. | Effects of dapagliflozin in combination with metoprolol sustained-release tablets on prognosis and cardiac function in patients with acute myocardial infarction after PCI | |
| WO2021073249A1 (en) | USE OF β-NMN IN PREPARATION OF DRUG FOR TREATING AND PREVENTING SEPSIS-INDUCED ORGAN DAMAGE | |
| Cui et al. | Minocycline attenuates oxidative and inflammatory injury in a intestinal perforation induced septic lung injury model via down-regulating lncRNA MALAT1 expression | |
| Bao et al. | Artemisinin and its derivate alleviate pulmonary hypertension and vasoconstriction in rodent models | |
| Zhang et al. | Hydrogen regulates mitochondrial quality to protect glial cells and alleviates sepsis-associated encephalopathy by Nrf2/YY1 complex promoting HO-1 expression | |
| Wróbel et al. | SN003, a CRF1 receptor antagonist, attenuates depressive-like behavior and detrusor overactivity symptoms induced by 13-cis-retinoic acid in rats | |
| Feng et al. | Protective roles of hydroxyethyl starch 130/0.4 in intestinal inflammatory response and survival in rats challenged with polymicrobial sepsis | |
| Xiao et al. | Sanggenon C protects against pressure overload‑induced cardiac hypertrophy via the calcineurin/NFAT2 pathway | |
| Hu et al. | Sevoflurane postconditioning improves the spatial learning and memory impairments induced by hemorrhagic shock and resuscitation through suppressing IRE1α-caspase-12-mediated endoplasmic reticulum stress pathway | |
| CN109369754B (en) | Nitrate ester compound and application thereof | |
| CN115337300A (en) | Use of water-soluble prodrugs of acacetin for the effective treatment of pulmonary hypertension | |
| WO2021093376A1 (en) | Use of phosphodiesterase 5 inhibitor in preparation of medicament for resisting fibrotic diseases | |
| Gao et al. | SUMO2-mediated SUMOylation of SH3GLB1 promotes ionizing radiation-induced hypertrophic cardiomyopathy through mitophagy activation |
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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20230301 Address after: Room 1002, Building C, Talent Building, No. 10, Xinghuo Road, Mount Taishan Street, Jiangbei New District, Nanjing, Jiangsu 210031 Applicant after: Nanjing anmaohua Pharmaceutical Co.,Ltd. Address before: 710000 East half of the first floor of Building 3, Zhengkun Scientific Innovation Industrial Base, No. 10, Xingyuan Road, Fengjing Industrial Park, Heyi District, Xi'an, Shaanxi Applicant before: Xi'an Baikangning Biological Products Co.,Ltd. |
|
| TA01 | Transfer of patent application right |