TWI556818B - 哺乳類sirt1的活化劑 - Google Patents
哺乳類sirt1的活化劑 Download PDFInfo
- Publication number
- TWI556818B TWI556818B TW104125781A TW104125781A TWI556818B TW I556818 B TWI556818 B TW I556818B TW 104125781 A TW104125781 A TW 104125781A TW 104125781 A TW104125781 A TW 104125781A TW I556818 B TWI556818 B TW I556818B
- Authority
- TW
- Taiwan
- Prior art keywords
- psoralen
- sirt1
- yeast
- mammalian
- mammalian sirt1
- Prior art date
Links
- 239000012190 activator Substances 0.000 title description 8
- ZCCUUQDIBDJBTK-UHFFFAOYSA-N psoralen Chemical compound C1=C2OC(=O)C=CC2=CC2=C1OC=C2 ZCCUUQDIBDJBTK-UHFFFAOYSA-N 0.000 claims description 80
- 108010041191 Sirtuin 1 Proteins 0.000 claims description 41
- VXGRJERITKFWPL-UHFFFAOYSA-N 4',5'-Dihydropsoralen Natural products C1=C2OC(=O)C=CC2=CC2=C1OCC2 VXGRJERITKFWPL-UHFFFAOYSA-N 0.000 claims description 40
- 150000001875 compounds Chemical class 0.000 claims description 16
- 230000004913 activation Effects 0.000 claims description 8
- 201000010099 disease Diseases 0.000 claims description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 3
- 208000008589 Obesity Diseases 0.000 claims description 3
- 235000020824 obesity Nutrition 0.000 claims description 3
- 238000011144 upstream manufacturing Methods 0.000 claims description 3
- 102000000344 Sirtuin 1 Human genes 0.000 claims 4
- 102100031455 NAD-dependent protein deacetylase sirtuin-1 Human genes 0.000 description 37
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 26
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 23
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 18
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 12
- 238000000605 extraction Methods 0.000 description 11
- 238000001262 western blot Methods 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 101000654471 Mus musculus NAD-dependent protein deacetylase sirtuin-1 Proteins 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 229960003966 nicotinamide Drugs 0.000 description 6
- 235000005152 nicotinamide Nutrition 0.000 description 6
- 239000011570 nicotinamide Substances 0.000 description 6
- 108091006473 SLC25A33 Proteins 0.000 description 5
- 102100033827 Solute carrier family 25 member 33 Human genes 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 4
- 230000003213 activating effect Effects 0.000 description 4
- XDROKJSWHURZGO-UHFFFAOYSA-N angelicin Chemical compound C1=C2OC=CC2=C2OC(=O)C=CC2=C1 XDROKJSWHURZGO-UHFFFAOYSA-N 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 201000005202 lung cancer Diseases 0.000 description 4
- 208000020816 lung neoplasm Diseases 0.000 description 4
- 201000008968 osteosarcoma Diseases 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical group NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 3
- 230000021736 acetylation Effects 0.000 description 3
- 238000006640 acetylation reaction Methods 0.000 description 3
- 230000006196 deacetylation Effects 0.000 description 3
- 238000003381 deacetylation reaction Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 229930030856 Psoralidin Natural products 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
- 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
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 101150018117 cobB gene Proteins 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 206010012601 diabetes mellitus Diseases 0.000 description 2
- SWDSVBNAMCDHTF-UHFFFAOYSA-N isowighteone Chemical compound C1=C(O)C(CC=C(C)C)=CC(C=2C(C3=C(O)C=C(O)C=C3OC=2)=O)=C1 SWDSVBNAMCDHTF-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229950006238 nadide Drugs 0.000 description 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- MLMVLVJMKDPYBM-UHFFFAOYSA-N pseudoisopsoralene Natural products C1=C2C=COC2=C2OC(=O)C=CC2=C1 MLMVLVJMKDPYBM-UHFFFAOYSA-N 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 229940016667 resveratrol Drugs 0.000 description 2
- 235000021283 resveratrol Nutrition 0.000 description 2
- 108010048734 sclerotin Proteins 0.000 description 2
- 101150089009 sir2 gene Proteins 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 210000005253 yeast cell Anatomy 0.000 description 2
- ORILYTVJVMAKLC-UHFFFAOYSA-N Adamantane Natural products C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 description 1
- 102100022983 B-cell lymphoma/leukemia 11B Human genes 0.000 description 1
- LFYJSSARVMHQJB-UHFFFAOYSA-N Backuchiol Natural products CC(C)=CCCC(C)(C=C)C=CC1=CC=C(O)C=C1 LFYJSSARVMHQJB-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- PWAACAMQKVIVPZ-UHFFFAOYSA-N Corylin Natural products OC1=CC=C2C(=O)C(C3=CC=C4OC(C=CC4=C3)(C)C)=COC2=C1 PWAACAMQKVIVPZ-UHFFFAOYSA-N 0.000 description 1
- 108010058643 Fungal Proteins Proteins 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 102100039990 Hairy/enhancer-of-split related with YRPW motif protein 2 Human genes 0.000 description 1
- 102100038885 Histone acetyltransferase p300 Human genes 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101000903697 Homo sapiens B-cell lymphoma/leukemia 11B Proteins 0.000 description 1
- 101001002170 Homo sapiens Glutamine amidotransferase-like class 1 domain-containing protein 3, mitochondrial Proteins 0.000 description 1
- 101001035089 Homo sapiens Hairy/enhancer-of-split related with YRPW motif protein 2 Proteins 0.000 description 1
- 101000882390 Homo sapiens Histone acetyltransferase p300 Proteins 0.000 description 1
- 101001123331 Homo sapiens Peroxisome proliferator-activated receptor gamma coactivator 1-alpha Proteins 0.000 description 1
- 101000843556 Homo sapiens Transcription factor HES-1 Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 101000978776 Mus musculus Neurogenic locus notch homolog protein 1 Proteins 0.000 description 1
- 102000003945 NF-kappa B Human genes 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 108010016731 PPAR gamma Proteins 0.000 description 1
- 102100038825 Peroxisome proliferator-activated receptor gamma Human genes 0.000 description 1
- 102100028960 Peroxisome proliferator-activated receptor gamma coactivator 1-alpha Human genes 0.000 description 1
- 244000226566 Psoralea corylifolia Species 0.000 description 1
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 1
- 241000239226 Scorpiones Species 0.000 description 1
- 102000011990 Sirtuin Human genes 0.000 description 1
- 108050002485 Sirtuin Proteins 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 102100030798 Transcription factor HES-1 Human genes 0.000 description 1
- 108010078814 Tumor Suppressor Protein p53 Proteins 0.000 description 1
- 102000015098 Tumor Suppressor Protein p53 Human genes 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- LFYJSSARVMHQJB-GOSISDBHSA-N bakuchinol Natural products CC(C)=CCC[C@@](C)(C=C)C=CC1=CC=C(O)C=C1 LFYJSSARVMHQJB-GOSISDBHSA-N 0.000 description 1
- LFYJSSARVMHQJB-QIXNEVBVSA-N bakuchiol Chemical compound CC(C)=CCC[C@@](C)(C=C)\C=C\C1=CC=C(O)C=C1 LFYJSSARVMHQJB-QIXNEVBVSA-N 0.000 description 1
- 229940117895 bakuchiol Drugs 0.000 description 1
- KXXXNMZPAJTCQY-UHFFFAOYSA-N bakuchiol Natural products CC(C)CCCC(C)(C=C)C=Cc1ccc(O)cc1 KXXXNMZPAJTCQY-UHFFFAOYSA-N 0.000 description 1
- 235000020827 calorie restriction Nutrition 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000013058 crude material Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical group CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 101150046266 foxo gene Proteins 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229940101270 nicotinamide adenine dinucleotide (nad) Drugs 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 229910052707 ruthenium Inorganic materials 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- IASPBORHOMBZMY-UHFFFAOYSA-N srt1720 Chemical compound C=1N=C2C=CC=CC2=NC=1C(=O)NC1=CC=CC=C1C(N=C1SC=2)=CN1C=2CN1CCNCC1 IASPBORHOMBZMY-UHFFFAOYSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/487—Psoralea
-
- 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
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Diabetes (AREA)
- Alternative & Traditional Medicine (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Emergency Medicine (AREA)
- Cardiology (AREA)
- Endocrinology (AREA)
- Hospice & Palliative Care (AREA)
- Toxicology (AREA)
- Heart & Thoracic Surgery (AREA)
- Psychiatry (AREA)
- Child & Adolescent Psychology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
本發明關於一種活化劑，且特別攸關一種哺乳類SIRT1的活化劑。
SIRT1（哺乳類）為sirtuin家族的成員之一。SIRT1為一種菸醯胺腺嘌呤二核苷酸（nicotinamide adenine dinucleotide，NAD）依賴的脫乙醯酶（deacetylase），其可移除多數組織蛋白（histone）與非組織蛋白的乙醯基，因而涉及於許多生理現象，如：基因表現的控制、代謝、老化。具體而言，SIRT1自菸醯胺腺嘌呤二核苷酸移除菸醯胺（nicotinamide）後，將基質的乙醯基轉移至經菸醯胺移除的菸醯胺腺嘌呤二核苷酸。SIRT1所反應的基質仍持續增加中，已知包含多種轉錄因子，如腫瘤抑制蛋白p53、FoxO家族的成員、HES1、HEY2、PPARγ、CTIP2、p300、PGC-1α、NF-κB。
直到今日，已知的SIRT1活化劑相當稀少，如Lamin A、白藜蘆醇（resveratrol）、SRT1720、SRT2379。於是，確實有必要開發更多種的SIRT1活化劑，以期應用於涉及的生理現象。
本發明之一目的在於提出一種如式（1）所示化合物的用途，此用途為用於製備哺乳類SIRT1的活化劑，其中式（1）為：
…（1）。
本發明之另一目的在於提出一種如式（1）所示化合物的用途，此用途為用於製備延長壽命的藥劑，其中式（1）為：
…（1）。
本發明之又一目的在於提出一種如式（1）所示化合物的用途，此用途為用於製備與哺乳類SIRT1活化有關之疾病的藥劑，其中式（1）為：
…（1）。
為讓本發明上述及/或其他目的、功效、特徵更為明顯易懂，下文特舉較佳實施方式，作詳細說明：
本發明係基於一項不可思議之發現所完成的，其中萃取自補骨脂（
Psoralea corylifolia）的式（1）所示化合物能增進酵母菌壽命。根據此發現，進一步發現此化合物尚能活化哺乳類SIRT1。須敘明的是，式（1）為：
…（1）； 式（1）所示化合物於學術上亦稱作「補骨脂寧」或「榛仁球蛋白」（corylin）。若無特別強調，本文所使用的「式（1）所示化合物」、「補骨脂寧」、或「榛仁球蛋白」用語代表相同意義。
本發明的第一實施方式提出一種補骨脂寧的用途，其為用於製備哺乳類SIRT1的活化劑。補骨脂寧能藉由活化哺乳類SIRT1的上游來活化哺乳類SIRT1。此外，補骨脂寧可利用化學合成或於自然界萃取等方式取得，如自補骨脂萃取。
如Cell Metab. 2013; 18(3): 416-30所述，SIRT1的活化能延長小鼠的壽命。如前所述，由於補骨脂寧可活化哺乳類SIRT1，故其可延長壽命。於是，本發明的第二實施方式提出一種補骨脂寧的用途，其為用於製備延長壽命的藥劑。下文的實施例已證實補骨脂寧可延長酵母菌壽命。具體地說，補骨脂寧可製作成藥劑以投予至有此需求的個體。同樣地，補骨脂寧可活化哺乳類SIRT1來達到延長壽命的功效。同樣地，補骨脂寧可採用化學合成或於自然界萃取等方式取得，如自補骨脂萃取。
如Future Cardiol. 2012; 8(1): 89-100所述，白藜蘆醇可透過活化哺乳類SIRT1而治療心血管疾病。又如Cell Metab. 2012; 16(2): 180-8所述，脂肪組織內哺乳類SIRT1的失活會造成肥胖。再如Cell. 2010; 142(2): 320-32所述，於小鼠內SIRT1可透過活化ADAM 10α-分泌酶（secretase）來抑制阿茲海默症。另如Nature. 2007; 450(7170): 712-6所述，活化哺乳類SIRT1可作為治療糖尿病的標的。如前所述，補骨脂寧因可活化哺乳類SIRT1，故其可治療與哺乳類SIRT1活化有關的疾病。依此，本發明的第三實施方式提出一種補骨脂寧的用途，其為用於製備與哺乳類SIRT1活化有關之疾病的藥劑。本實施方式的疾病可以為但不限於心血管疾病、肥胖、阿茲海默症、或糖尿病。也就是說，補骨脂寧可作成藥劑來投予至有此需求的個體。同樣地，補骨脂寧可透過活化哺乳類SIRT1來達到治療上述疾病的功效。同樣地，補骨脂寧可採用化學合成或於自然界萃取等方式取得，如自補骨脂萃取。
茲以下述實施例，例示說明本發明以上實施方式：
＜補骨脂寧的萃取＞
請參照圖1，粉碎5.4公斤的補骨脂種子後，用11公升的乙醇浸泡萃取，此包含冷浸4次、70℃熱萃4小時5次。過濾萃取產物後，濃縮得到的乙醇抽粗物。接著，用正己烷與水分配萃取乙醇抽粗物而取得正己烷層與水層。水層再以乙酸乙酯進行分配萃取而得到乙酸乙酯層與水層。而正己烷層通入矽膠管柱後，利用不同濃度比例的正己烷/乙酸乙酯混合液沖提以依序分離得11個分層。
第1個分層為以50：1的正己烷/乙酸乙酯混合液沖提得到的，含補骨脂酚（bakuchiol）；第3個分層為以20：1的正己烷/乙酸乙酯混合液沖提得到的，含異補骨脂素（isopsoralen）；第4個分層為以7：1的正己烷/乙酸乙酯混合液沖提得到的，含補骨脂素（psoralen）；第5個分層為以5：1的正己烷/乙酸乙酯混合液沖提得到的，含補骨脂寧；第7個分層為以1：1的正己烷/乙酸乙酯混合液沖提得到的，含補骨脂次素（psoralidin）。
＜補骨脂寧對酵母菌壽命的延長＞
挑出單一酵母菌落至YEPD培養液中，並均勻混合。於菌液OD值達0.6至0.8時，取20μl菌液至含不同濃度藥劑的YEPD固態培養基，並採用三向顯微定位系統取出20至30個酵母菌細胞。於30℃下培養酵母菌細胞3至5小時使其分離出第0代子細胞。接著，於30℃下培養第0代子細胞3至5小時使其分離出第1代子細胞。重複上述步驟，直到未有任何代數的子細胞分離出。最後，依據分離次數便可計算出酵母菌的代數。
如圖2所示，野生型酵母菌的代數較補骨脂寧處理之野生型酵母菌的代數少（26.9 vs. 33.2），而
sir2突變酵母菌的代數較補骨脂寧處理之
sir2突變酵母菌的代數少（9.8 vs. 10.6）。意謂著補骨脂寧能延長酵母菌的壽命。又如圖3所示，補骨脂寧的有效濃度約為2至100μM，較佳地為2至15μM。
＜補骨脂寧對酵母菌
SIR2的活化＞
酵母菌
SIR2與哺乳類SIRT1互為同源，且咸知酵母菌
SIR2與延長壽命有關（Genes Dev. 2000; 14(9): 1021-6）。於此，將確認補骨脂寧延長壽命的效果是否與酵母菌
SIR2有關。
首先，於補骨脂寧處理酵母菌不同時間後，萃取酵母菌的RNA。接著，利用反轉錄聚合酶連鎖反應取得酵母菌RNA所對應的cDNA。最後，以聚合酶連鎖反應與即時聚合酶連鎖反應分析得到的cDNA。如圖4A、4B所示，補骨脂寧可隨著處理時間的增加來促進
PNC1的轉錄。
為了進一步驗證上述結果，於以不同濃度補骨脂寧處理酵母菌不同時間後，萃取酵母菌的蛋白質，並利用西方點墨法分析。如圖4C所示，與經卡路里限制（calorie restriction）10小時的酵母菌比較，可看出50μM補骨脂寧處理10小時的酵母菌可大量地表現Pnc1。表示說補骨脂寧可促進
PNC1的轉譯。
Pnc1為酵母菌Sir2的上游，因此推測補骨脂寧可經由活化Pnc1來活化酵母菌Sir2。
＜補骨脂寧對哺乳類SIRT1的活化＞
於此，將確認補骨脂寧是否會活化哺乳類SIRT1。
將人類骨肉瘤細胞U2OS於37℃下培養至隔夜後，投予補骨脂寧至細胞，再於37℃下培養。經不同的培養時間後，萃取細胞的蛋白質並以西方點墨法分析。
如圖5A所示，補骨脂寧可隨著培養時間的增加來促進哺乳類SIRT1的表現。
另外，將人類肺癌細胞A549或人類骨肉瘤細胞U2OS於37℃下培養至隔夜後，先添加含不同濃度藥劑的培養液至細胞，再於37℃下預先培養4小時。然後，利用UV光照射細胞以乙醯基化p53，並於37℃下繼續培養4小時。最後，萃取細胞的蛋白質並以西方點墨法分析。
如圖5B、5C所示，補骨脂寧可抑制p53的乙醯基化。依「先前技術」所述，p53為哺乳類SIRT1的受質。此結果表示補骨脂寧可提升哺乳類SIRT1之下游受質的去乙醯基化；亦即，補骨脂寧可活化哺乳類SIRT1的下游受質。
菸醯胺已知為哺乳類SIRT1的抑制劑（Exp Hematol. 2012; 40(4): 342-55），於此利用此化合物進一步驗證上述結果。先將人類肺癌細胞A549於37℃下培養隔夜後，加入含菸醯胺的培養液至細胞，並於37℃下培養1小時。之後，加入不同濃度的補骨脂寧至培養液內，且於37℃下培養細胞4小時。然後，利用UV光照射細胞以乙醯基化p53後，於37℃下繼續培養4小時。最後，萃取細胞蛋白質，並以西方點墨法分析。
如圖6所示，菸醯胺可抑制補骨脂寧活化哺乳類SIRT1所造成的p53去乙醯基化。此結果可進一步證實補骨脂寧所造成的p53去乙醯基化須哺乳類SIRT1的參與。
綜上所陳，補骨脂寧可活化哺乳類SIRT1，故補骨脂寧具有作為哺乳類SIRT1活化劑的潛力。
惟以上所述者，僅為本發明之較佳實施例，但不能以此限定本發明實施之範圍；故，凡依本發明申請專利範圍及發明說明書內容所作之簡單的等效改變與修飾，皆仍屬本發明專利涵蓋之範圍內。
無
圖1為一流程圖，說明補骨脂寧自補骨脂種子的萃取。 圖2為一曲線圖，說明補骨脂寧處理之酵母菌的壽命。 圖3為一曲線圖，說明不同濃度補骨脂寧處理之酵母菌的壽命。 圖4A為一聚合酶連鎖反應電泳圖，說明補骨脂寧於不同時間處理之酵母菌的
PNC1轉錄。 圖4B為一即時聚合酶連鎖反應結果圖，說明補骨脂寧於不同時間處理之酵母菌的
PNC1轉錄。 圖4C為一西方點墨法結果圖，說明補骨脂寧於不同時間處理之酵母菌的
PNC1轉譯。 圖5A為一西方點墨法結果圖，說明補骨脂寧於不同時間處理之人類骨肉瘤細胞U2OS的哺乳類SIRT1表現。 圖5B為一西方點墨法結果圖，說明不同濃度補骨脂寧於不同時間處理之人類肺癌細胞A549的p53乙醯基化。 圖5C為一西方點墨法結果圖，說明不同濃度補骨脂寧於不同時間處理之人類骨肉瘤細胞U2OS的p53乙醯基化。 圖6為一西方點墨法結果圖，說明菸醯胺處理之人類肺癌細胞A549的p53去乙醯基化。
Claims (9)
- 一種如式(1)所示化合物的用途，係用於製備活化哺乳類SIRT1的製劑，其中該式(1)為：
- 如請求項第1項所述之用途，其中該化合物為萃取自補骨脂。
- 如請求項第1項所述之用途，其中該化合物活化該哺乳類SIRT1的上游。
- 一種如式(1)所示化合物的用途，係用於製備延長壽命的藥劑，其中該式(1)為：
- 如請求項第4項所述之用途，其中該化合物為萃取自補骨脂。
- 如請求項第4項所述之用途，其中該化合物活化哺乳類SIRT1來延長壽命。
- 一種如式(1)所示化合物的用途，係用於製備與哺乳類SIRT1活化有關之疾病的藥劑，其中該式(1)為：
- 如請求項第7項所述之用途，其中該化合物為萃取自補骨脂。
- 如請求項第7項所述之用途，其中該疾病為心血管疾病或肥胖。
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW104125781A TWI556818B (zh) | 2015-08-07 | 2015-08-07 | 哺乳類sirt1的活化劑 |
| CN201510670027.6A CN105343051A (zh) | 2015-08-07 | 2015-10-13 | 补骨脂宁于制备哺乳类sirt1活化剂、与其活化有关的疾病的药剂、延寿药剂的用途 |
| US14/885,645 US20170035724A1 (en) | 2015-08-07 | 2015-10-16 | Mammalian sirt1 activator |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW104125781A TWI556818B (zh) | 2015-08-07 | 2015-08-07 | 哺乳類sirt1的活化劑 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| TWI556818B true TWI556818B (zh) | 2016-11-11 |
| TW201705951A TW201705951A (zh) | 2017-02-16 |
Family
ID=55319216
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| TW104125781A TWI556818B (zh) | 2015-08-07 | 2015-08-07 | 哺乳類sirt1的活化劑 |
Country Status (3)
| Country | Link |
|---|---|
| US (1) | US20170035724A1 (zh) |
| CN (1) | CN105343051A (zh) |
| TW (1) | TWI556818B (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106619605A (zh) * | 2017-01-22 | 2017-05-10 | 中国药科大学 | 补骨脂宁的医药用途 |
| CN110090208B (zh) * | 2019-04-24 | 2021-11-23 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | 补骨脂宁在制备Nrf2抑制剂、与Nrf2抑制有关的疾病的药物、抗癌药物的应用 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101257897A (zh) * | 2005-07-07 | 2008-09-03 | 西特里斯药业公司 | 用于治疗或预防肥胖、胰岛素抵抗障碍和线粒体相关障碍的方法和相关组合物 |
| MX2013011996A (es) * | 2011-04-15 | 2013-11-01 | Nestec Sa | Metodos para regular la expresion de genes de sirtuina. |
| JP6364484B2 (ja) * | 2013-07-17 | 2018-07-25 | ザ ホン コン ユニバーシティ オブ サイエンス アンド テクノロジイ | 神経保護剤としてのＥｐｈＡ４阻害剤 |
-
2015
- 2015-08-07 TW TW104125781A patent/TWI556818B/zh active
- 2015-10-13 CN CN201510670027.6A patent/CN105343051A/zh active Pending
- 2015-10-16 US US14/885,645 patent/US20170035724A1/en not_active Abandoned
Non-Patent Citations (1)
| Title |
|---|
| Journal of Natural Products (20150424) Vol. 78, No. 4, pp. 666-673 * |
Also Published As
| Publication number | Publication date |
|---|---|
| TW201705951A (zh) | 2017-02-16 |
| US20170035724A1 (en) | 2017-02-09 |
| CN105343051A (zh) | 2016-02-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Li et al. | Luteolin protects against diabetic cardiomyopathy by inhibiting NF-κB-mediated inflammation and activating the Nrf2-mediated antioxidant responses | |
| Feng et al. | Polysaccharides extracted from Phellinus linteus ameliorate high-fat high-fructose diet induced insulin resistance in mice | |
| Kim et al. | The inhibition of JNK MAPK and NF-κB signaling by tenuifoliside A isolated from Polygala tenuifolia in lipopolysaccharide-induced macrophages is associated with its anti-inflammatory effect | |
| Vo et al. | Down-regulation of histamine-induced endothelial cell activation as potential anti-atherosclerotic activity of peptides from Spirulina maxima | |
| CN105377289A (zh) | 用于下调Bcl-xL和/或Bcl-w的活性和/或量的试剂 | |
| Sedky et al. | The molecular basis of cytotoxicity of α‐spinasterol from Ganoderma resinaceum: Induction of apoptosis and overexpression of p53 in breast and ovarian cancer cell lines | |
| Huang et al. | Anti-viral effect of a compound isolated from Liriope platyphylla against hepatitis B virus in vitro | |
| Li et al. | Fatty liver mediated by peroxisome proliferator‐activated receptor‐α DNA methylation can be reversed by a methylation inhibitor and curcumin | |
| Kim et al. | Autophagy and microRNA dysregulation in liver diseases | |
| Zhai et al. | Cross-talk of MicroRNA and hydrogen sulfide: A novel therapeutic approach for bone diseases | |
| Su et al. | The function of LncRNA-H19 in cardiac hypertrophy | |
| TWI556818B (zh) | 哺乳類sirt1的活化劑 | |
| Yu et al. | MIR-190B alleviates cell autophagy and burn-induced skeletal muscle wasting via modulating PHLPP1/Akt/FoxO3A signaling pathway | |
| Cheng et al. | Pleiotropic ameliorative effects of ellagitannin geraniin against metabolic syndrome induced by high-fat diet in rats | |
| Huang et al. | NBM-HD-3, a novel histone deacetylase inhibitor with anticancer activity through modulation of PTEN and AKT in brain cancer cells | |
| Lee et al. | Effects of magnolialide isolated from the leaves of Laurus nobilis L.(Lauraceae) on immunoglobulin E-mediated type I hypersensitivity in vitro | |
| Yu et al. | Cardio-protective and Anti-atherosclerosis Effect of Crocetin on Vitamin D3 and HFD-induced Atherosclerosis in Rats | |
| Han et al. | In vitro anti-tumor activity in SGC-7901 human gastric cancer cells treated with dandelion extract | |
| Ngo et al. | Protective effects of peptides from skate (Okamejei kenojei) skin gelatin against endothelial dysfunction | |
| Lam et al. | Latex C‐serum from H evea brasiliensis induces non‐apoptotic cell death in hepatocellular carcinoma cell line (HepG2) | |
| Li et al. | Protective effects of Nrf2 against sepsis-induced hepatic injury | |
| Jeevanandam et al. | Biological macromolecules as antidiabetic agents | |
| Bai et al. | Aloin Attenuates Oxidative Stress, Inflammation, and CCl4-Induced Liver Fibrosis in Mice: Possible Role of TGF-β/Smad Signaling | |
| Zhang et al. | 3, 4-Dihydroxyphenylethanol ameliorates lipopolysaccharide-induced septic cardiac injury in a murine model | |
| CN110139658A (zh) | 组合物 |