CN108721627B - 类泛素化抑制剂在制备预防或治疗骨质疏松症的药物中的用途 - Google Patents
类泛素化抑制剂在制备预防或治疗骨质疏松症的药物中的用途 Download PDFInfo
- Publication number
- CN108721627B CN108721627B CN201810359098.8A CN201810359098A CN108721627B CN 108721627 B CN108721627 B CN 108721627B CN 201810359098 A CN201810359098 A CN 201810359098A CN 108721627 B CN108721627 B CN 108721627B
- Authority
- CN
- China
- Prior art keywords
- inhibitor
- mln4924
- ubiquitination
- subject
- bone
- 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.)
- Active
Links
- 239000003112 inhibitor Substances 0.000 title claims abstract description 51
- 208000001132 Osteoporosis Diseases 0.000 title claims abstract description 45
- 239000003814 drug Substances 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 102000044159 Ubiquitin Human genes 0.000 title description 5
- 108090000848 Ubiquitin Proteins 0.000 title description 5
- MPUQHZXIXSTTDU-QXGSTGNESA-N sulfamic acid [(1S,2S,4R)-4-[4-[[(1S)-2,3-dihydro-1H-inden-1-yl]amino]-7-pyrrolo[2,3-d]pyrimidinyl]-2-hydroxycyclopentyl]methyl ester Chemical compound C1[C@H](O)[C@H](COS(=O)(=O)N)C[C@H]1N1C2=NC=NC(N[C@@H]3C4=CC=CC=C4CC3)=C2C=C1 MPUQHZXIXSTTDU-QXGSTGNESA-N 0.000 claims abstract description 82
- 230000034512 ubiquitination Effects 0.000 claims abstract description 37
- 238000010798 ubiquitination Methods 0.000 claims abstract description 37
- 230000037396 body weight Effects 0.000 claims description 16
- 241000282414 Homo sapiens Species 0.000 claims description 10
- 230000009527 neddylation Effects 0.000 claims description 2
- 210000002997 osteoclast Anatomy 0.000 abstract description 54
- 230000004069 differentiation Effects 0.000 abstract description 30
- 210000000988 bone and bone Anatomy 0.000 abstract description 22
- 210000004027 cell Anatomy 0.000 description 45
- 230000000694 effects Effects 0.000 description 39
- 108020004707 nucleic acids Proteins 0.000 description 25
- 102000039446 nucleic acids Human genes 0.000 description 25
- 150000007523 nucleic acids Chemical class 0.000 description 25
- 102000003436 UBA3 Human genes 0.000 description 19
- 108060008744 UBA3 Proteins 0.000 description 19
- 241000699666 Mus <mouse, genus> Species 0.000 description 18
- 230000002401 inhibitory effect Effects 0.000 description 16
- 102000038427 NEDD8-activating enzyme E1 Human genes 0.000 description 15
- 108091007790 NEDD8-activating enzyme E1 Proteins 0.000 description 15
- 108020004459 Small interfering RNA Proteins 0.000 description 15
- 238000000034 method Methods 0.000 description 15
- 206010065687 Bone loss Diseases 0.000 description 13
- 241000699670 Mus sp. Species 0.000 description 13
- 210000002540 macrophage Anatomy 0.000 description 13
- 239000002924 silencing RNA Substances 0.000 description 13
- 238000011282 treatment Methods 0.000 description 13
- 208000006386 Bone Resorption Diseases 0.000 description 12
- 230000015572 biosynthetic process Effects 0.000 description 12
- 230000024279 bone resorption Effects 0.000 description 12
- 102000014128 RANK Ligand Human genes 0.000 description 11
- 108010025832 RANK Ligand Proteins 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 10
- 230000002358 autolytic effect Effects 0.000 description 10
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 10
- 229940079593 drug Drugs 0.000 description 10
- 229940088598 enzyme Drugs 0.000 description 10
- 210000000689 upper leg Anatomy 0.000 description 10
- 206010017076 Fracture Diseases 0.000 description 9
- 102000007591 Tartrate-Resistant Acid Phosphatase Human genes 0.000 description 9
- 108010032050 Tartrate-Resistant Acid Phosphatase Proteins 0.000 description 9
- 238000003786 synthesis reaction Methods 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 8
- 238000010186 staining Methods 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- 238000010171 animal model Methods 0.000 description 7
- 238000009806 oophorectomy Methods 0.000 description 7
- 230000004044 response Effects 0.000 description 7
- 230000001225 therapeutic effect Effects 0.000 description 7
- 210000002303 tibia Anatomy 0.000 description 7
- 208000010392 Bone Fractures Diseases 0.000 description 6
- 101710190440 Cytotoxin 1 Proteins 0.000 description 6
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 6
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 239000006143 cell culture medium Substances 0.000 description 6
- VYVRIXWNTVOIRD-LRHBOZQDSA-N ciguatoxin CTX1B Chemical compound C([C@@]12[C@@H](C)[C@@H]([C@@H]3[C@H]([C@H]([C@H](C)[C@H]4O[C@H]5C[C@@H](C)C[C@H]6O[C@@]7(C)[C@H](O)C[C@H]8O[C@H]9C=C[C@H]%10O[C@H]%11C[C@@H]%12[C@H]([C@@H]([C@H]%13O[C@H](C=CC[C@@H]%13O%12)\C=C\[C@H](O)CO)O)O[C@@H]%11C=C[C@@H]%10O[C@@H]9C\C=C/C[C@@H]8O[C@@H]7C[C@@H]6O[C@@H]5C[C@@H]4O3)O)O2)C)[C@H](O)CO1 VYVRIXWNTVOIRD-LRHBOZQDSA-N 0.000 description 6
- 239000002679 microRNA Substances 0.000 description 6
- 210000000963 osteoblast Anatomy 0.000 description 6
- 230000037361 pathway Effects 0.000 description 6
- 208000029725 Metabolic bone disease Diseases 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 206010049088 Osteopenia Diseases 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- -1 methoxyethyl Chemical group 0.000 description 5
- 230000011164 ossification Effects 0.000 description 5
- 239000002243 precursor Substances 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 229940011871 estrogen Drugs 0.000 description 4
- 239000000262 estrogen Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 108091070501 miRNA Proteins 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- 102000002574 p38 Mitogen-Activated Protein Kinases Human genes 0.000 description 4
- 108010068338 p38 Mitogen-Activated Protein Kinases Proteins 0.000 description 4
- 230000026731 phosphorylation Effects 0.000 description 4
- 238000006366 phosphorylation reaction Methods 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 229950003937 tolonium Drugs 0.000 description 4
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 description 4
- 206010006002 Bone pain Diseases 0.000 description 3
- 102000055006 Calcitonin Human genes 0.000 description 3
- 108060001064 Calcitonin Proteins 0.000 description 3
- 108010022452 Collagen Type I Proteins 0.000 description 3
- 102000012422 Collagen Type I Human genes 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- 101000939348 Homo sapiens NEDD8-activating enzyme E1 regulatory subunit Proteins 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- 102100029781 NEDD8-activating enzyme E1 regulatory subunit Human genes 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 108091027967 Small hairpin RNA Proteins 0.000 description 3
- 230000037182 bone density Effects 0.000 description 3
- 229960004015 calcitonin Drugs 0.000 description 3
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 3
- 230000003197 catalytic effect Effects 0.000 description 3
- 239000012930 cell culture fluid Substances 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 125000001153 fluoro group Chemical group F* 0.000 description 3
- 239000007928 intraperitoneal injection Substances 0.000 description 3
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 229920002477 rna polymer Polymers 0.000 description 3
- 239000004055 small Interfering RNA Substances 0.000 description 3
- 238000003325 tomography Methods 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- JKYKXTRKURYNGW-UHFFFAOYSA-N 3,4-dihydroxy-9,10-dioxo-9,10-dihydroanthracene-2-sulfonic acid Chemical compound O=C1C2=CC=CC=C2C(=O)C2=C1C(O)=C(O)C(S(O)(=O)=O)=C2 JKYKXTRKURYNGW-UHFFFAOYSA-N 0.000 description 2
- TZTRUHFXPVXWRD-QTQZEZTPSA-N 4-amino-7-[(2R,3R,4S,5R)-3,4-dihydroxy-5-[(sulfamoylamino)methyl]oxolan-2-yl]-5-[2-(2-ethoxy-6-fluorophenyl)ethynyl]pyrrolo[2,3-d]pyrimidine Chemical compound NC=1C2=C(N=CN=1)N(C=C2C#CC1=C(C=CC=C1F)OCC)[C@@H]1O[C@@H]([C@H]([C@H]1O)O)CNS(N)(=O)=O TZTRUHFXPVXWRD-QTQZEZTPSA-N 0.000 description 2
- 229940122361 Bisphosphonate Drugs 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 101800001415 Bri23 peptide Proteins 0.000 description 2
- 101800000655 C-terminal peptide Proteins 0.000 description 2
- 102400000107 C-terminal peptide Human genes 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000282693 Cercopithecidae Species 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- 206010020100 Hip fracture Diseases 0.000 description 2
- 102100031911 NEDD8 Human genes 0.000 description 2
- 108700004934 NEDD8 Proteins 0.000 description 2
- 101150107958 NEDD8 gene Proteins 0.000 description 2
- 241001597008 Nomeidae Species 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 101100532088 Oryza sativa subsp. japonica RUB2 gene Proteins 0.000 description 2
- 101100532090 Oryza sativa subsp. japonica RUB3 gene Proteins 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 208000027790 Rib fracture Diseases 0.000 description 2
- 208000006011 Stroke Diseases 0.000 description 2
- 108010049264 Teriparatide Proteins 0.000 description 2
- 102000004243 Tubulin Human genes 0.000 description 2
- 108090000704 Tubulin Proteins 0.000 description 2
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 2
- 229930003316 Vitamin D Natural products 0.000 description 2
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 230000009102 absorption Effects 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229940117893 apigenin Drugs 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 239000000090 biomarker Substances 0.000 description 2
- 150000004663 bisphosphonates Chemical class 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 210000002805 bone matrix Anatomy 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical group C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000007402 cytotoxic response Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 238000001361 intraarterial administration Methods 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 230000004072 osteoblast differentiation Effects 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 230000004962 physiological condition Effects 0.000 description 2
- 238000013001 point bending Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000003449 preventive effect Effects 0.000 description 2
- 230000000069 prophylactic effect Effects 0.000 description 2
- 229960004622 raloxifene Drugs 0.000 description 2
- GZUITABIAKMVPG-UHFFFAOYSA-N raloxifene Chemical compound C1=CC(O)=CC=C1C1=C(C(=O)C=2C=CC(OCCN3CCCCC3)=CC=2)C2=CC=C(O)C=C2S1 GZUITABIAKMVPG-UHFFFAOYSA-N 0.000 description 2
- PZSJYEAHAINDJI-UHFFFAOYSA-N rhodium(3+) Chemical compound [Rh+3] PZSJYEAHAINDJI-UHFFFAOYSA-N 0.000 description 2
- 101150024074 rub1 gene Proteins 0.000 description 2
- 229940079488 strontium ranelate Drugs 0.000 description 2
- XXUZFRDUEGQHOV-UHFFFAOYSA-J strontium ranelate Chemical compound [Sr+2].[Sr+2].[O-]C(=O)CN(CC([O-])=O)C=1SC(C([O-])=O)=C(CC([O-])=O)C=1C#N XXUZFRDUEGQHOV-UHFFFAOYSA-J 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- OGBMKVWORPGQRR-UMXFMPSGSA-N teriparatide Chemical compound C([C@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)[C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 OGBMKVWORPGQRR-UMXFMPSGSA-N 0.000 description 2
- 229960005460 teriparatide Drugs 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 2
- 235000019166 vitamin D Nutrition 0.000 description 2
- 239000011710 vitamin D Substances 0.000 description 2
- 150000003710 vitamin D derivatives Chemical class 0.000 description 2
- 229940046008 vitamin d Drugs 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- HJCMDXDYPOUFDY-WHFBIAKZSA-N Ala-Gln Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CCC(N)=O HJCMDXDYPOUFDY-WHFBIAKZSA-N 0.000 description 1
- 208000000044 Amnesia Diseases 0.000 description 1
- 208000006820 Arthralgia Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 1
- 101100262441 Caenorhabditis elegans rfl-1 gene Proteins 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 208000005189 Embolism Diseases 0.000 description 1
- 206010014733 Endometrial cancer Diseases 0.000 description 1
- 206010014759 Endometrial neoplasm Diseases 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 108010088842 Fibrinolysin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 108060003393 Granulin Proteins 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 101000644669 Homo sapiens NEDD8-conjugating enzyme Ubc12 Proteins 0.000 description 1
- 235000008586 Hovenia Nutrition 0.000 description 1
- 241000405398 Hovenia Species 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010020850 Hyperthyroidism Diseases 0.000 description 1
- 208000013038 Hypocalcemia Diseases 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- 208000000913 Kidney Calculi Diseases 0.000 description 1
- 206010023509 Kyphosis Diseases 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 208000002720 Malnutrition Diseases 0.000 description 1
- 101000830602 Mus musculus Tumor necrosis factor ligand superfamily member 11 Proteins 0.000 description 1
- 208000000112 Myalgia Diseases 0.000 description 1
- LOJFGJZQOKTUBR-XAQOOIOESA-N NC(N)=NCCC[C@@H](C(O)=O)NC(=O)CNC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(O)=O)C)CC1=CN=CN1 Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)CNC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCC(O)=O)C)CC1=CN=CN1 LOJFGJZQOKTUBR-XAQOOIOESA-N 0.000 description 1
- 102100020710 NEDD8-conjugating enzyme Ubc12 Human genes 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029148 Nephrolithiasis Diseases 0.000 description 1
- 102100034404 Nuclear factor of activated T-cells, cytoplasmic 1 Human genes 0.000 description 1
- 101710151542 Nuclear factor of activated T-cells, cytoplasmic 1 Proteins 0.000 description 1
- 208000003076 Osteolysis Diseases 0.000 description 1
- 206010031264 Osteonecrosis Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 108090000445 Parathyroid hormone Proteins 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 239000012083 RIPA buffer Substances 0.000 description 1
- 108091030071 RNAI Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 108091081021 Sense strand Proteins 0.000 description 1
- 208000005392 Spasm Diseases 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 231100000480 WST assay Toxicity 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 206010000059 abdominal discomfort Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 108010009896 bone resorption factor Proteins 0.000 description 1
- 230000037118 bone strength Effects 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 125000003636 chemical group Chemical group 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000036569 collagen breakdown Effects 0.000 description 1
- 108010049937 collagen type I trimeric cross-linked peptide Proteins 0.000 description 1
- 238000003271 compound fluorescence assay Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000003235 crystal violet staining Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000011461 current therapy Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 229960001251 denosumab Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000009504 deubiquitination Effects 0.000 description 1
- BVQAWSJMUYMNQN-UHFFFAOYSA-N dipyridophenazine Chemical compound C1=CC=C2C3=NC4=CC=CC=C4N=C3C3=CC=CN=C3C2=N1 BVQAWSJMUYMNQN-UHFFFAOYSA-N 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 201000006828 endometrial hyperplasia Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002834 estrogen receptor modulator Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 230000009368 gene silencing by RNA Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 230000001744 histochemical effect Effects 0.000 description 1
- 230000000705 hypocalcaemia Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 208000029791 lytic metastatic bone lesion Diseases 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 230000001071 malnutrition Effects 0.000 description 1
- 235000000824 malnutrition Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000009245 menopause Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 150000004702 methyl esters Chemical class 0.000 description 1
- FIYXUOWXHWJDAM-UHFFFAOYSA-N methyl sulfamate Chemical compound COS(N)(=O)=O FIYXUOWXHWJDAM-UHFFFAOYSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 210000001721 multinucleated osteoclast Anatomy 0.000 description 1
- 208000013465 muscle pain Diseases 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 208000015380 nutritional deficiency disease Diseases 0.000 description 1
- 230000001599 osteoclastic effect Effects 0.000 description 1
- 208000008798 osteoma Diseases 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 239000000199 parathyroid hormone Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229940075559 piperine Drugs 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 230000036316 preload Effects 0.000 description 1
- 108091007428 primary miRNA Proteins 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 229940126409 proton pump inhibitor Drugs 0.000 description 1
- 239000000612 proton pump inhibitor Substances 0.000 description 1
- 238000002601 radiography Methods 0.000 description 1
- 230000009103 reabsorption Effects 0.000 description 1
- 230000030233 regulation of protein ubiquitination Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000010948 rhodium Substances 0.000 description 1
- 230000005586 smoking cessation Effects 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
-
- 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
- A61K31/12—Ketones
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/555—Heterocyclic compounds containing heavy metals, e.g. hemin, hematin, melarsoprol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
- C12N2310/141—MicroRNAs, miRNAs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/50—Physical structure
- C12N2310/53—Physical structure partially self-complementary or closed
- C12N2310/531—Stem-loop; Hairpin
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Genetics & Genomics (AREA)
- Physical Education & Sports Medicine (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Rheumatology (AREA)
- General Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
本发明内容是关于类泛素化抑制剂的用途,其是用以制备一药物,以预防一个体罹患骨质疏松症,或是治疗一罹患或疑似罹患骨质疏松症的个体。该类泛素化抑制剂可抑制破骨细胞的分化及功效,进而增加骨骼的生物力学特性。依据本发明内容某些实施方式,该类泛素化抑制剂是MLN4924。
Description
技术领域
本发明是关于一种类泛素化(neddylation)抑制剂的新用途。更具体来说,本发明是关于利用类泛素化抑制剂来制备预防一个体罹患骨质疏松症或治疗一罹患骨质疏松症的个体的药物。
背景技术
骨质疏松症(osteoporosis)是一种好发于老年人的骨骼代谢异常疾病,女性的发生率约为男性的3-5倍。临床上,罹患骨质疏松症的病患会因骨质流失导致骨质减少,轻微的压力即有可能使其发生骨折(常见包含髋骨骨折、脊椎骨折、腕骨骨折及肋骨骨折等)及骨疼痛等症状。已知停经中年妇女会因体内***(estrogen)的减少而促使骨质快速流失。此外,包含酒精、咖啡因、吸烟、维生素D摄取不足、营养不良、重金属(例如镉或铅)接触、甲状腺功能亢进、肾脏疾病及特定治疗(例如某些抗癌药物、质子泵抑制剂及类固醇)等因素皆有可能影响个体的骨质含量。
除了传统射线摄影术(radiography)外,目前亦可利用双能X光吸收检测(Dual-energy X-ray absorptiometry,DXA)及生物标志分析来诊断骨质疏松症。在双能X光吸收检测中,若个体的骨质密度(T分数,T-score) 大于等于-1.0,代表其具有正常骨质含量;当个体的骨质密度介于-1.0 及-2.5之间时,代表其骨质不足(osteopenia);而当个体的骨密度低于或等于-2.5时,则代表其罹患骨质疏松症。生物标志分析则是通过检测诸如细胞自溶酶K(cathepsin K,可分解骨质中第I型胶原蛋白的酶)、新表位(neoepitope,骨质疏松所导致的骨碎片)及C-端肽(C-telopeptide,第I 型胶原蛋白分解产物)等标的分子于个体体内的含量来得知个体的骨质分解的状况。
已知戒烟及减少酒精的摄取皆有助于减少骨质的流失。除此之外,亦有报导指出增加维生素D或钙质的摄取量可降低个体罹患骨质不足及 /或骨质疏松症的机率。在药物方面,目前包含双膦酸酯类 (bisphosphonate,一种可抑制骨质流失的药物)、***(estrogen,一种女性荷尔蒙,可降低骨质流失)、特立帕肽(Teriparatide,重组副甲状腺素,可刺激骨质合成)、雷洛昔芬(Raloxifene,***受器调节剂,可抑制骨骼的分解代谢)、狄诺塞麦(Denosumab,核因子κ-B配体的受体活化剂(Receptor activator ofnuclear factorκ-B ligand(RANKL))的抑制剂)、降钙素(Calcitonin,可抑制破骨细胞(osteoclast)作用)及雷奈酸锶 (strontium ranelate,可同时抑制骨质疏松及促进骨质生成的双效药物)等药物皆可通过减少骨流失失来达到预防/治疗骨质疏松症的疗效。然而,这些药物皆有其临床限制,而无法提供个体安全且有效预防/治疗功效。举例来说,双膦酸酯类会产生腹痛、恶心、消化不良、骨痛、关节痛、肌肉疼痛及颚骨坏死等副作用;***会造成静脉栓塞、心肌梗塞、中风、乳癌、子宫内膜增生及子宫内膜癌等问题;特立帕肽除了注射剂量高且价格昂贵之外,亦会引发痉挛、头晕及恶性骨瘤等病状;雷洛昔芬会增加个体血栓及中风的机率;狄诺塞麦除了会造成低血钙及颚骨坏死之外,亦会影响个体免疫调节,使其易于遭受微生物感染;降钙素会导致过敏及癌症的产生;至于雷奈酸锶,短期服用会造成个体肠胃道不适及头痛等症状,长期服用则会引发个体意识障碍、甚至记忆丧失等严重副作用。
有鉴于此,相关领域亟需一种新的方法,可有效且安全地预防/治疗罹患骨质疏松症,以提升有罹患骨质疏松症风险的个体及骨质疏松症病患的生活质量及寿命。
发明内容
发明内容旨在提供本申请内容的简化摘要,以使阅读者对本发明内容具备基本的理解。此发明内容并非本申请内容的完整概述,且其用意并非在指出本发明实施例的重要/关键组件或界定本发明的范围。
本发明内容是关于一种类泛素化抑制剂的用途,其是用于制备一药物以预防一个体罹患骨质疏松症,或是治疗一罹患骨质疏松症的个体。
因此,本发明内容的第一方面是关于一种用以预防一个体罹患骨质疏松症或治疗一罹患骨质疏松症的个体的方法。该方法包含对该个体投予一有效量的类泛素化抑制剂。
依据本发明内容某些实施方式,该类泛素化抑制剂是一种NEDD8 活化酶(NEDD8-activating enzyme,NAE)抑制剂。依据较佳的实施方式,该NAE抑制剂是一种化合物或合成核酸。示例性的NAE抑制物包含 MLN4924及其类似物(analog)或衍生物(derivative)、TAS4464、6,6”-联芹菜甙元(6,6”-biapigenin)、环金属化的铑(III)复合物(cyclometallated rhodium(III)complex,[Rh(ppy)2(dppz)]+)及卡瓦胡椒素A(flavokawain A)。依据本发明内容一实施方式,该NAE抑制剂是MLN4924。依据本发明内容另一实施方式,该NAE抑制剂是一合成核酸,其包含SEQ.ID NO.:4或6的序列,或是与SEQ.IDNO.:4或6的序列相对应的DNA 序列。在该实施方式中,合成核酸可抑制UBA3(NAE的催化次单元)的表达量。
该合成核酸可以是小干扰核糖核酸(small interference ribonucleic acids,siRNAs)、小发夹核糖核酸(small hairpin ribonucleic acids,shRNAs) 或微核糖核酸(micro-ribonucleic acids,miRNAs)。依据本发明内容一操作实施例,该合成核酸是siRNA。
依据本发明内容某些实施方式,该个体为一小鼠,其中类泛素化抑制剂的投予有效量为每日每公斤个体体重0.125-1,250毫克;较佳的,投予有效量为每日每公斤个体体重1.25-125毫克;更佳的,投予有效量为每日每公斤个体体重1.25-25毫克。依据一操作实施方式,投予至小鼠的类泛素化抑制剂的有效量为每公斤体重10毫克。
本领域技术人员可依据动物模式的使用剂量来计算本发明类泛素化抑制剂的人体等效剂量(human equivalent dose,HED)。据此,人类的有效量为每日每公斤体重0.01-100毫克;较佳为0.1-10毫克;最佳为0.1-2 毫克。
依据本发明内容实施方式,该类泛素化抑制剂的投予次数为每日一次,且连续投予至少一周;较佳为每日投予一次,且连续投予至少二周,以达抑制骨质疏松的效果。
本发明类泛素化抑制剂可经由口服、肠内、鼻腔、局部、黏膜及非口服方式投予至个体体内,其中非口服方式可以是皮下、真皮内、肌肉内、动脉内、静脉内、脊椎内或腹腔内注射。
本发明内容另一方面是关于一合成核酸,其包含SEQ.ID NO.:4或 6的序列,或与SEQ.ID NO.:4或6的序列相对应的DNA序列,其中合成核酸可抑制UBA3(NAE的催化次单元)的表达量。该合成核酸可以是siRNA、shRNA或miRNA。
非必要性地,该合成核酸的5’端及/或3’端具有氟基(fluoro group)、甲氧基(methoxy group)或甲氧基乙基(methoxyethyl group)修饰。
在参阅下文实施方式后,本领域技术人员当可轻易了解本发明的基本精神及其它发明目的,以及本发明所采用的技术手段与实施方式。
附图说明
为让本发明的上述与其它目的、特征、优点与实施例能更明显易懂,附图的说明如下:
图1A-1D为依据实施例1.1所阐述的数据,其中是将源自小鼠骨髓的巨噬细胞(marrow-derived macrophage,BMM)培养于包含sRANKL (soluble RANKL,一种重组RANKL;浓度为每毫升50奈克)、巨噬细胞群落刺激因子(macrophage colony-stimulating factor(M-CSF);每毫升40 奈克)及特定浓度的MLN4924的细胞培养液,7天后进行后续分析;图1A:利用TRAP染色来评估巨噬细胞分化成巨大多核破骨细胞的结果;图1B:于人工骨质基质上进行破骨细胞分化后,移除细胞,再利用1%甲苯胺蓝分析再吸收区域,以确认破骨细胞吸收骨质的活性;图1C:收集细胞培养液以分析细胞自溶酶K的活性;图1D:利用MTT试验来确定特定浓度的MLN4924对BMM细胞所产生的细胞毒性;比例尺为100 微米;将数据表示为平均值±标准差;*表示p<0.05,**表示p<0.01;
图2A-2D为依据实施例1.2所阐述的数据,其中是将RAW264.7细胞培养于包含sRANKL(每毫升50奈克)及特定浓度的MLN4924的细胞培养液,7天后进行后续分析;图2A:利用TRAP染色来评估破骨细胞的分化结果;图2B:于人工骨质基质上进行破骨细胞分化后,移除细胞,再利用1%甲苯胺蓝分析再吸收区域,藉以确认破骨细胞吸收骨质的活性;图2C:收集细胞培养液以分析细胞自溶酶K的活性;图2D:利用 MTT试验来确定特定浓度的MLN4924对RAW264.7巨噬细胞所产生的细胞毒性;比例尺为100微米;将数据表示为平均值±标准差;*表示p< 0.05;
图3A-3D为依据实施例2所阐述的数据,其中是对巨噬细胞投予特定浓度的MLN4924处理30分钟后,以sRANKL(每毫升100奈克)刺激细胞,并于特定时间点收集细胞进行西方墨点分析;图3A:(A) MLN4924于特定时间点对ERK、P38与JNK磷酸化的影响;图3B:不同浓度的MLN4924对ERK、P38与JNK磷酸化的影响;图3C:MLN4924 于特定时间点对NFATc-1表达量的影响;图3D:不同浓度的MLN4924 对NFATc-1表达量的影响;对照组:未处理;R:sRANKL;M:MLN4924;
图4A-4B为依据实施例3所阐述的数据,其中是将小鼠的成骨前趋细胞株(pre-osteoblast)MC3T3-E1培养于包含不同浓度的MLN4924的细胞培养液,21天后进行后续分析;图4A:利用结晶紫染色来分析于成骨作用(osteogenesis)过程中MC3T3-E1的细胞生长情况;图4B:以茜素红S(Alizarin Red S)染色来定量分析MC3T3-E1细胞于第21天的成骨作用;diff:成骨细胞分化培养液;MLN:MLN4924;
图5A-5D为依据本发明内容实施例4所绘示的数据,其分别阐述破骨细胞分化时,sRANKL对类泛素化路径的作用,以及,UBA3siRNA 对巨噬细胞的抑制功效。图5A:将RAW264.7小鼠巨噬细胞种植于培养盘后,投予不同浓度的sRANKL(每毫升0-100奈克)处理。72小时后,溶解细胞并利用西方墨点法以特定抗体进行分析。图5B:将RAW264.7 小鼠巨噬细胞种植于培养盘后,以UBA3或对照siRNA(scrambled siRNA)转染细胞。培养后,收集、溶解并以西方墨点法分析这些经转染的细胞中UBA3的表达量。图5C及5D:以sRANKL(每毫升50奈克) 处理经UBA3或对照siRNA转染的RAW264.7细胞;培养3天后,拍照记录并计算经TRAP染色的巨噬细胞的外观改变。将数据表示为平均值±标准差,*p<0.05。
图6A-6E为依据实施例5.1所阐述的数据,其中是对小鼠进行卵巢切除术后,投予sRANKL(每公斤体重1毫克),以产生骨质疏松症的动物模式;图6A:经特定处理的小鼠远程股骨的微电脑断层扫瞄二维照片,左图为轴向平面,右图为冠状结构平面;图6B-6D:利用微电脑断层扫瞄来测量小鼠胫骨的小梁骨(trabecular bone)的骨质参数,包含骨体积/ 总体积百分比(图6B)、小梁骨数量(图6C)及小梁骨间隙(图6D);图6E:对照组、OVX、OVX+sRANKL及OVX+sRANKL+MLN4924组小鼠血清中CTX-1的表达量;将数据表示为平均值±标准差;每组只数为6;* 代表p<0.05,**代表p<0.01;
图7A-7D为依据实施例5.2所阐述的数据,是关于对照组、OVX、 OVX+RANKL及OVX+RANKL+MLN4924组小鼠的股骨的力学特性;图7A:包含模数(modulus);图7B:断裂应变(Strain at Break);图7C:降伏压力(Stress at Yield);图7D:断裂负载(Load atBreak);将数据表示为平均值±标准差;每组只数为6;*代表p<0.05,**代表p<0.01。
具体实施方式
为了使本发明内容的叙述更加详尽与完备,下文针对了本发明的实施方式与具体实施例提出了说明性的描述;但这并非实施或运用本发明具体实施例的唯一形式。实施方式中涵盖了多个具体实施例的特征以及用以建构与操作这些具体实施例的方法步骤与顺序。然而,亦可利用其它具体实施例来达成相同或均等的功能与步骤顺序。
虽然用以界定本发明较广范围的数值范围与参数皆是约略的数值,此处已尽可能精确地呈现具体实施例中的相关数值。然而,任何数值本质上不可避免地含有因个别测试方法所致的标准偏差。在此处,“约”通常是指实际数值在一特定数值或范围的正负10%、5%、1%或0.5%之内。或者是,“约”一词代表实际数值落在平均值的可接受标准误差之内,视本领域技术人员的考虑而定。除了实验例之外,或除非另有明确的说明,当可理解此处所用的所有范围、数量、数值与百分比(例如用以描述材料用量、时间长短、温度、操作条件、数量比例及其它相似者)均经过“约”的修饰。因此,除非另有相反的说明,本说明书与附随申请专利范围所揭示的数值参数皆为约略的数值,且可视需求而更动。至少应将这些数值参数理解为所指出的有效位数与套用一般进位法所得到的数值。在此处,将数值范围表示成由一端点至另一段点或介于二端点之间;除非另有说明,此处所述的数值范围皆包含端点。
除非本说明书另有定义,此处所用的科学与技术词汇的含义与本领域技术人员所理解与惯用的意义相同。此外,在不和上下文冲突的情形下,本说明书所用的单数名词涵盖该名词的复数型;而所用的复数名词时亦涵盖该名词的单数型。
除非本说明书另有定义,否则本文中核苷酸的标记方式依循一般原则,即左手端是5’-端,而右手端为3’-端。
在本发明内容中,“MLN4924的类似物”(analogs of MLN4924)一词是指这些源自MLN4924,且保留MLN4924全部或部分生物功能的化合物、对映异构体(enantiomer)及衍生物。
在本发明内容中,“MLN4924的衍生物”(derivatives of MLN4924)一词是指这些源自MLN4924,且保留MLN4924全部或部分生物功能的化合物、对映异构体(enantiomer)及类似物。
“-“应用”(application)或“投予”(administration)在此为可互换的词汇,皆是用以阐述将本发明的类泛素化抑制剂(例如MLN4924)提供至一个体(例如有罹患骨质疏松症的风险的个体,或是罹患/疑似罹患骨质疏松症的个体),以降低该个体罹患骨质疏松症的风险,或是治疗、改善个体骨质疏松症的症状。依据本发明内容不同的实施方式,可通过不同路径 (例如静脉内注射或腹腔内注射)将本发明的类泛素化抑制剂传送至个体体内,据以达到预防/治疗骨质疏松症的目的。
在本发明内容中,“治疗”(treat,treating or treatment)一词是指部份或完全预防、改善、减轻及/或处理罹患骨质疏松症的个体的相关病征,其中是通过抑制破骨细胞的生成及/或作用来使罹患骨质疏松症的个体的相关病征获得改善。示例性的与骨质疏松症相关的病征包含,但不限于,骨折(例如髋骨骨折、脊椎骨折、腕骨骨折及肋骨骨折)、骨疼痛、驼背、肠胃不适、便秘、呼吸困难、血钙增加及肾结石。“治疗”(treat,treating ortreatment)一词于此说明书中亦指应用或施予本发明内容的类泛素化抑制剂(例如MLN4924)至一患有骨质疏松症的个体身上,以达到部份或完全减轻、减缓、治愈疾病、延迟发病、抑制病程发展、降低疾病严重性,及/或降低骨质疏松症的发生。
“-“有效量”(effective amount)在此处是指一药物的用量足以产生欲求的疗效反应。具体的有效量取决于多种因素,如欲治疗的特定状况、患者的生理条件(如,患者体重、年龄或性别)、接受治疗的哺乳动物或动物的类型、治疗持续时间、目前疗法(如果有的话)的本质以及所用的具体配方和化合物或其衍生物的结构。有效量亦指一种化合物或组合物,其治疗利益效果超越其毒性或有害影响。举例来说,可将有效量表示成药物的总重量(譬如以克、毫克或微克为单位)或表示成药物重量与体重的比例(其单位为毫克/公斤(mg/kg))。技术人员可依据动物模式的剂量来计算药物(如本发明内容的类泛素化抑制剂)的人体等效剂量(human equivalent dose,HED)。举例来说,技术人员可依据美国食品药物管理局 (US Food and Drug Administration,FDA)所公告的“估算成人健康志愿者在初始临床治疗测式的最大安全起始剂量”(Estimating the Maximum Safe Starting Dose inInitial Clinical Trials for Therapeutics in Adult Healthy Volunteers)来估算人体使用的最高安全剂量。
“-“个体”(subject)一词是指包含人类的动物,其可接受本发明方法的治疗。除非特定指出,否则“个体”(subject)一词同时意指男性及女性。依据本发明内容较佳实施方式,所述个体为一哺乳动物,包括但不限于,人类、小鼠、大鼠、猴子、猪、狗、猫、马、羊、牛及兔子等。在一特定实施例中,该个体为人类。
本发明内容部分是基于发明人首度发现类泛素化抑制剂具有抑制破骨细胞生成及/或作用的新用途,据以提供一种用以预防或治疗骨质疏松症的方法。
依据本发明内容实施方式,本发明方法包含对一有需要的个体投予一有效量的类泛素化抑制剂。该类泛素化抑制剂可以是任何可抑制类泛素化反应的拮抗剂(例如类泛素化反应的拮抗性肽或化合物)。依据本发明内容某些实施方式,该类泛素化抑制剂是可专一地抑制NAE。该NAE 抑制剂可以是一化合物或一合成核酸。适合用于本发明方法的化合物包含,但不限于,MLN4924及其类似物及衍生物、TAS4464、6,6”-联芹菜甙元、环金属化的铑(III)复合物及卡瓦胡椒素A。
依据本发明内容较佳实施方式,类泛素化抑制剂是MLN4924或其类似物或衍生物,其中该MLN4924的全名为氨基磺酸 (1S,2S,4R)-4-(4-((1S)-2,3-二氢-1H-茚-1-基氨)-7H-吡咯(2,3-d)嘧啶-7- 基)-2-羟环戊基)甲酯((1S,2S,4R)-4-(4-((1S)-2,3-Dihydro-1H- inden-1-ylamino)-7H-pyrrolo(2,3-d)pyrimidin-7-yl)-2-hydroxycyclopentyl)methyl sulphamate)。
至于合成核酸,其可以是一siRNA、一shRNA或一miRNA。依据本发明内容某些实施方式,该合成核酸是siRNA。在一较佳实施方式中,该siRNA具有一作为传递链(passengerstrand)的正链(sense strand),以及一作为引导股(guide strand)以静默基因表现的负链(anti-sense strand),其中该正链具有SEQ.ID NO.:3的序列,且该负链具有SEQ.IDNO.:4的序列。依据另一实施例,该正链具有SEQ.ID NO.:5的序列,且该负链具有SEQ.IDNO.:6的序列。
如相关领域习知技艺人士所知,可以通过siRNAs以外的核苷酸分子来静默或抑制mRNA的转译,而这些核苷酸分子亦涵盖于本发明内容。举例来说,shRNA为一种RNA分子,其是通过一小段核苷酸所形成的间距(spacer)来连结正链及负链,因而形成一环形(loop)结构,且依据本发明内容的实施方式,第一、第二及第三核酸可以是shRNAs,其中这些shRNAs的正链分别等同于SEQ.ID NO.:3或5的序列,而其负链则等同于SEQ.ID NO.:4或6的序列。在其它实施方式中,核酸是以 miRNA或其前驱物(例如,pri-miRNA或pre-miRNA)的形式提供,且 miRNA或其前驱物分别具有一包含SEQ.ID NO.:4或6的序列。亦或者,本核酸可以是任何双股或单股的负链寡核苷酸(oligonucleotides),其分别包含SEQ.ID NO.:4或6的序列(或与SEQ.ID NO.:4或6相对应的DNA序列)。
可通过不同化学基团的修饰来增加合成核酸的稳定性、延长合成核酸的半衰期、增加合成核酸的功能或使合成核酸标的至特定组织/细胞。举例来说,可以氟基(-F)、甲氧基(-OCH3,O-Me)或甲氧基乙基 (-OCH2CH2OCH3,O-MOE)来修饰合成核酸的5’端及/或3’端。具体来说,为增加合成核酸的稳定性,可以2′-O-甲基(2′-O-Me)、2′-氟基(2′-F)或2′- 甲氧基乙基(2′-O-MOE)来置换合成核酸的核糖的2′-OH基团。此外或亦或是,可利用胆固醇或硫醇基来修饰合成核酸的5’端或3端,藉以增加合成核酸的传递及细胞穿透功效。当可想见,合成核酸的5’端或3’端可与荧光分子等分子键结。
利用本发明方法预防或治疗的个体可以是一有罹患骨质不足或骨质疏松症的风险的个体、一罹患或疑似罹患骨质疏松症的个体,或是一罹患或疑似罹患骨质不足的个体。
依据本发明内容某些实施方式,接受本发明方法预防或治疗的个体是一小鼠。在这些实施方式中,类泛素化抑制剂的投予有效量为每日每公斤个体体重0.125-1,250毫克;较佳为每日每公斤个体体重1.25-125 毫克;更佳为每日每公斤个体体重1.25-25毫克。依据本发明内容一操作实施例,投予至个体的类泛素化抑制剂的有效量为每日每公斤个体体重10毫克。
本领域技术人员可依据动物模式的使用剂量来计算本发明类泛素化抑制剂的人体等效剂量。据此,人类的有效量为每日每公斤体重 0.01-100毫克;例如每日每公斤体重0.01、0.02、0.03、0.04、0.05、0.06、 0.07、0.08、0.09、0.1、0.11、0.12、0.13、0.14、0.15、0.16、0.17、0.18、 0.19、0.2、0.21、0.22、0.23、0.24、0.25、0.26、0.27、0.28、0.29、0.3、0.31、0.32、0.33、0.34、0.35、0.36、0.37、0.38、0.39、0.4、0.41、0.42、 0.43、0.44、0.45、0.46、0.47、0.48、0.49、0.5、0.51、0.52、0.53、0.54、 0.55、0.56、0.57、0.58、0.59、0.6、0.61、0.62、0.63、0.64、0.65、0.66、 0.67、0.68、0.69、0.7、0.71、0.72、0.73、0.74、0.75、0.76、0.77、0.78、 0.79、0.8、0.81、0.82、0.83、0.84、0.85、0.86、0.87、0.88、0.89、0.9、0.91、0.92、0.93、0.94、0.95、0.96、0.97、0.98、0.99、1、1.1、1.2、 1.3、1.4、1.5、1.6、1.7、1.8、1.9、2、2.1、2.2、2.3、2.4、2.5、2.6、 2.7、2.8、2.9、3、3.1、3.2、3.3、3.4、3.5、3.6、3.7、3.8、3.9、4、4.1、 4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5、5.1、5.2、5.3、5.4、5.5、 5.6、5.7、5.8、5.9、6、6.1、6.2、6.3、6.4、6.5、6.6、6.7、6.8、6.9、7、 7.1、7.2、7.3、7.4、7.5、7.6、7.7、7.8、7.9、8、8.1、8.2、8.3、8.4、 8.5、8.6、8.7、8.8、8.9、9、9.1、9.2、9.3、9.4、9.5、9.6、9.7、9.8、 9.9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、 25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、 41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、 57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、 73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、 89、90、91、92、93、94、95、96、97、98、99或100毫克;较佳为0.1-10毫克;最佳为0.1-2毫克。
临床操作人员可依据使用需求的不同来调整投予剂量、投予频率及投予次数;举例来说,依据个体年龄、骨质疏松的程度、生理状况、其它治疗(若有)及性别每周投予1-7次类泛素化抑制剂(例如每周投予1、2、 3、4、5、6或7次类泛素化抑制剂);且连续投予1、2、3、4或更多周。或者是,可每2周、每4周、每5周、每6周、每7周、每8周、每9 周或每10周对个体投予1-10次(例如1、2、3、4、5、6、7、8、9或10 次)类泛素化抑制剂;或是于每1个月、每2个月、每3个月或更长的时间内对个体投予1次类泛素化抑制剂。依据本发明内容一实施例,是对个体每日投予类泛素化抑制剂,且连续投予至少2周,以达到预防/ 治疗效果;举例来说,连续投予2、3、4或更多周。
依据本发明内容一实施方式,本发明方法可通过抑制类泛素化路径来抑制破骨细胞的生成及/或作用,进而达到预防或治疗骨质疏松症的功效。依据本发明内容另一实施方式,本发明方法不会显著影响成骨细胞 (osteoblast)的存活及骨质生成作用。
当可想见,本发明类泛素化抑制剂可单独投予至个体体内来预防或治疗骨质疏松症。或者是,本发明类泛素化抑制剂可与另一对骨质疏松症具有预防/治疗功效的药剂共同投予至个体体内,据以增加预防/治疗功效。依据使用需求不同,本发明类泛素化抑制剂可于另一种药剂投予前、期间或之后投予至个体体内。
可任选地,本发明类泛素化抑制剂可经由肠内注射、口服、非口服或黏膜穿透方式投予至个体体内,其中非口服方式可以是皮下注射、静脉注射、动脉注射、脊椎注射或腹腔注射。
本发明内容所述的个体为一哺乳动物,举例来说,人类、小鼠、大鼠、猴子、猪、狗、猫、马、羊、牛及兔子等。在一特定实施例中,该个体为人类。
下文提出多个实验例来说明本发明的某些实施方式,以利本领域技术人员实现本发明,且不应将这些实验例视为对本发明范围的限制。据信技术人员在阅读了此处提出的说明后,可在不需过度解读的情形下,完整利用并实践本发明。此处所引用的所有公开文献,其全文皆视为本说明书的一部分。
实施例
材料及方法
试剂
由Active Biochem(Activebiochem,Maplewood,NJ)购买MLN4924。分别由CellGuidance Systems LLC(Carlsbad,CA)及R&D(R&D, Minneapolis,MN)购买重组小鼠RANKL(sRANKL,GFM4)及重组小鼠 M-CSF(416-ML)。由Santa Cruz Biotechnology(Santa Cruz,Santa Cruz, CA)购买NFATc1(7A6)抗体。由Cell Signaling technology(CST,Beverly,MA)购买pERK(4370)、ERK(4695)、pJNK(4668)、JNK(9252)、 p-p38-MAPK(4631)、p38-MAPK(9212)抗体。由GeneTex(GeneTex,Irvine, CA)购买GAPDH及α-微管蛋白(α-tubulin)抗体。
细胞培养及制备初级破骨细胞
将RAW264.7(小鼠单核球/巨噬细胞株)培养于包含10%胎牛血清 (Gibco)及1倍GlutaMax(Gibco)的高葡萄糖DMEM细胞培养液(Gibco BRL,Gaithersburg,MD)中,并置于包含5%CO2的环境中。分离源自小鼠骨髓的巨噬细胞(marrow-derived macrophage,BMM)以制备破骨前趋细胞(pre-osteoclast cell)。由6-12周大的C57BL/6J小鼠取其股骨及胫骨后,分离骨髓,据以制备破骨细胞。简单来说,将BMM细胞培养于每毫升40奈克的M-CSF培养液中3天,以制备单核球/巨噬细胞系的破骨前趋细胞。
形成破骨细胞
为于活体外形成破骨细胞,分别将破骨前趋细胞及RAW264.7培养于每毫升50奈克的sRANKL或不同浓度的MLN4924中7天,每两天换新的RANKL与MLN4924。接着,固定细胞,再依据使用者操作说明,以TRAP染色套组(KAMIYA Biomedical Company,Seattle,WA)染色分析抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)的表现。之后,利用Nikon TE300显微镜***(Nikon Instruments Inc.,Melville,NY) 观察及拍摄照片。
细胞自溶酶K活性试验及凹陷试验(pit assay)
将破骨前趋细胞及RAW264.7种植于Corning Osteo表面试验96-孔盘(Corning,Kennebunk,Maine)。24小时后,投予sRANKL(每毫升50 奈克)及不同浓度的MLN4924;每两天换新的RANKL与MLN4924,培养7天后,收集细胞,并以Biovision细胞自溶酶K活性荧光分析试验试剂盒(Biovision,Milpitas,CA)来分析细胞自溶酶K的活性。之后,以 1%甲苯胺蓝(toluidine blue)染孔盘底部骨盘,染色24小时,再以去离子水洗涤2次。利用Nikon TE300显微镜***,以100倍放大倍率(Nikon, Melville,NY)来拍摄照片。
西方墨点分析
将RAW264.7细胞以正常培养液种植在6-孔盘,隔天将培养液换成无血清的细胞培养液中,培养6小时后投予特定处理。在特定时间点时,以冰的磷酸盐缓冲液(phosphate-buffered saline.PBS)洗涤细胞后,利用包含完全蛋白酶抑制混合剂(Roche)的RIPA缓冲液溶解细胞。以SDS-PAGE 分离细胞溶解产物,之后以特定抗体进行西方墨点分析,这些抗体包含 NEDD8(2745;cell signaling),APPBP1(sc 360048;santa cruze)、UBA3(sc377272;santa cruze)及UBC12(sc 390064;santa cruze)。
UBA3抑制破骨细胞分化
将RAW264.7细胞种植于细胞培养盘后,以三种UBA3或对照siRNA (GenePharma)转染细胞,表1总结了UBA3siRNA 1(Uba3-MUS-110)、 UBA3siRNA 2(Uba3-MUS-574)、UBA3siRNA 3(Uba3-MUS-627)及对照siRNA(作为负对照组)的核苷酸序列。
表1用以抑制UBA表现的siRNA的核苷酸序列
转染6小时后,对细胞投予sRANKL(每毫升50奈克)。3天后,拍照记录并计算UBA3抑制对成熟巨大破骨细胞形成的影响。
动物模式
所有的实验流程皆于嘉义长庚纪念医院的实验动物照顾及使用委员会许可下进行,且符合中国 台湾地区科学管理部门的实验动物照顾及使用规范(IACUC号:2013100102及2015103001)。尽可能减少动物承受的痛苦。建立骨质疏松症的小鼠模式。简单来说,对C57BL/6J母鼠(6-8 周大;购买自中国 台湾乐斯科生物科技)进行卵巢切除术。手术48小时后,将sRANKL(每公斤体重1毫克)或生理食盐水腹腔注射至小鼠体内,每天注射一次,连续注射二天;之后以腹腔注射方式每天投予MLN4924(每公斤10毫克),共投予二周。以过量二氧化碳牺牲小鼠后,分离其胫骨及股骨。利用组织化学试验(histochemistry assay)分析胫骨;利用 MTS(Testresources,Shakopee,MN)对股骨进行弯曲试验,及利用Skyscan 1272(Bruker,Kontich,Belgium)进行断层扫描分析。以特定ELISA试剂盒 (USCN life scienceincorporation,Wuhan,Hubei,China)来检测血清中,第 I型胶原的交联C-端肽(CTX-1)(CEA665Mu)含量。
生物力学试验
以MTS Synergie 200(Testresources,Shakopee,MN)进行三点弯曲试验来检测股骨的力学特性。负载速度为每分钟1毫米,将预负载设定为 1N,将负载细胞设定为500N,将时间设定为30秒,且将二点之间的距离设定为10毫米。由包含模数、断裂负载、降伏压力及断裂应变等结构特性来绘制力-位移数据。
统计分析
将实验结果以平均值±平均标准偏差(Mean±Standard deviation)来表示。以非成对Student’s t试验来评估二组之间的差异性;当P值小于 0.05时,即视为二组间具有显著差异。
实施例1 MLN4924会抑制破骨细胞分化
实施例1将探讨MLN4924对由BMM及RAW264.7细胞所分化的破骨细胞的影响。结果分别阐述于图1-2。
1.1 BMM
将由小鼠的股骨及胫骨取出的初代BMM细胞培养于包含 sRANKL、M-CSF及特定浓度的MLN4924的细胞培养液中,以检测 MLN4924对破骨细胞生成的影响。经TRAP染色后,具有较大且多核的细胞为成熟的破骨细胞。图1A的TRAP染色结果显示,相较于未投予MLN4924处理的细胞,MLN4924可明显抑制破骨细胞的形成,且该抑制作用是与MLN4924的浓度相关。
为进一步确认MLN4924对破骨细胞活性的影响,接着以凹陷试验来分析破骨细胞的骨质再吸收活性。同样将小鼠的初代BMM细胞培养于包含sRANKL、M-CSF及特定浓度的MLN4924的细胞培养液中,7 天后利用1%甲苯胺蓝染色细胞,以观察再吸收区域。由图1B结果可知, MLN4924会以浓度相关的方式显著抑制破骨细胞的骨质再吸收活性。
已知在骨骼再吸收过程中,破骨细胞会分泌细胞自溶酶K来分解胶原蛋白及其它基质蛋白。因此本研究亦探讨MLN4924对细胞自溶酶K 的影响。分析结果显示,sRANKL会增加细胞自溶酶K的活性;相较于仅投予sRANKL的对照组,投予MLN4924可显著降低细胞自溶酶K的活性。此外,单独投予MLN4924并不会影响巨噬细胞中细胞自溶酶K 的基础活性(图1C)。
图1B及1C指出,MLN4924可通过抑制破骨细胞的分化及骨吸收活性来降低骨吸收异常。
接着,利用WST试验来分析上述观察到的抑制反应是否是因 MLN4924对BMM细胞产生毒杀反应所造成的结果。图1D显示,在可产生抑制反应的浓度下,MLN4924不会对细胞产生毒杀反应。
1.2 RAW264.7
除了小鼠的初代BMM外,本研究亦利用RAW264.7巨噬细胞株来确认MLN4924对破骨细胞的影响。
将RAW264.7细胞培养于包含sRANKL及特定浓度的MLN4924的细胞培养液中。7天后,利用TRAP染色来观察破骨细胞的分化结果。结果显示,MLN4924可显著抑制源自RAW264.7细胞(经sRANKL刺激活化)的破骨细胞的分化(图2A)、骨质再吸收活性(图2B)及细胞自溶酶K 活性(图2C)。此外,MLN4924不会影响RAW264.7细胞存活率的结果亦指出,MLN4924于可抑制破骨细胞分化的剂量下,不具有细胞毒杀作用(图2D)。
图2A-2D的结果指出,MLN4924在不影响巨噬细胞的存活率的情况下,可显著抑制破骨细胞的分化及其相关活性,进而预防骨质再吸收。
实施例2 MLN4924的作用机制
如图3A所示,在投予sRANKL后,磷酸化ERK(p-ERK)、磷酸化 p38(p-p38)及磷酸化JNK(p-JNK)的表达量会随着时间的增加而增加。相较于对照组(R组),sRANKL会明显增加ERK、p38与JNK的磷酸化程度(图3A);而投予MLN4924(R+M组)则可抑制经sRANKL刺激的ERK、p38与JNK的磷酸化程度(图3A)。图3B进一步证实,上述抑制效果会随着MLN4924浓度的增加而增加。
先前研究指出,RANKL会引发转录因子NFATc-1的表现,且由缺乏NFATc-1的胚胎所分离的干细胞无法成功分化为破骨细胞。此外,于破骨前趋细胞异源表现NFATc-1,可使其在无RANKL存在的情况下,分化为破骨细胞。有鉴于NFATc-1于破骨细胞分化过程中扮演着重要的角色,本研究亦探讨MLN4924是否会改变NFATc-1于细胞中的表达量。如图3C所示,sRANKL会引发NFATc-1表现(R组),而投予MLN4924 (R+M组)则会明显抑制NFATc-1的表达量。图3D则进一步证实该抑制效果会随着MLN4924浓度的增加而增加。
上述结果指出,MLN4924会以时间及浓度相关的方式抑制RANKL 的下游反应来有效抑制破骨细胞分化及其活性。
实施例3 MLN4924对成骨细胞的影响
除了破骨细胞外,本实施例将检测MLN4924是否亦会影响(即增加或减少)成骨细胞的数量或生成反应。图4A-4B阐述这些结果。
将小鼠的成骨前趋细胞株MC3T3-E1培养于包含不同浓度的 MLN4924的成骨细胞分化培养液中,共培养21天。结果显示,投予 MLN4924并不会影响成骨细胞的存活量(图4A)及分化(图4B)。
由上述结果可知,MLN4924在可有效抑制破骨细胞分化的剂量下,不会影响成骨细胞的存活量及分化。
实施例4对UBA3具有专一性的siRNA可抑制破骨细胞的分化
RANK为肿瘤坏死因子受体家族(tumor necrosis factor receptor family)的一员,主要会表现于破骨细胞及其前趋细胞上。已知RANK与其可溶性配体(即sRANKL)的作用会调控骨质的再吸收作用。通过与破骨前趋细胞的RANK结合,RANKL可调控破骨细胞的分化、增生及存活。为了探讨类泛素化路径于骨质疏松的重要性及治疗作用,本实施例将分析类泛素化路径的蛋白表达量,以及抑制UBA3表现对由sRANKL 刺激的破骨细胞分化的影响。
首先分析不同剂量的sRANKL对类泛素化路径的活化及破骨细胞的分化的作用。图5A结果显示,在培养3天后,sRANKL会以剂量相关的方式增加蛋白上NEDD8的修饰。此外,类泛素化路径的起始酶(E1) 为NEDD8活化酶(NAE),其是由APPBP1及UBA3次单元所组成的异二聚体。结果显示,在破骨细胞分化过程中,sRANKL会显著增加催化次单元UBA3的表达量,而不会影响次单元APPBP1的表达;亦即,在由sRANKL刺激的破骨细胞分化过程中,NAE的次单元UBA3扮演着重要的调控角色。
为了确认UBA3在由sRANKL活化的破骨细胞分化过程中所产生的功效,将二种可标的UBA3mRNA的RNAi分子转染至RAW264.7细胞,以抑制UBA3的表达。培养后,收集经转染的RAW264.7细胞,确认 UBA3的表达,并进一步投予sRANKL以诱发破骨细胞分化。经西方墨点分析后,图5B的结果显示,UBA3siRNA 2及3可抑制UBA3的表达量,而不会影响APPBP1的表达量;相较之下,UBA3siRNA 1及对照 siRNA则不会影响任何蛋白的表达量。此外,图5C及5D则指出,对照 siRNA不会抑制由sRANKL刺激的破骨细胞的分化。UBA3siRNA 2及 3则会显著降低破骨细胞的分化数量。这些数据指出,在由sRANKL刺激的破骨细胞分化过程中,UBA3的表达及活性于NAE媒介的蛋白类泛素化调控中扮演着重要的角色。
图5A-5D的结果证实对UBA3具有专一性的siRNA可抑制破骨细胞的分化。
实施例5 MLN4924于活体内的作用
5.1骨质流失
本实施例利用骨质疏松小鼠动物模式来分析MLN4924是否可通过抑制骨质再吸收异常来抑制破骨细胞的分化及活性,进而于活体产生治疗骨质疏松症的功效。
对小鼠投予特定处理后,利用微电脑断层扫描来分析其股骨的骨质状态。如图6A所示,相较于对照组,在投予卵巢切除术的小鼠(OVX组) 及进行卵巢切除术后给予sRANKL的小鼠(OVX+sRANKL组)的股骨皆可观察到严重的骨质流失;而投予MLN4924(OVX+sRANKL+MLN组) 则可明显减少骨质流失的情况。
此外,本研究亦分析小鼠胫骨的各种骨质参数,包含相对于总组织体积的相对于总体积的骨体积百分比(骨体积/总体积百分比,图6B)、小梁骨数目(图6C)及小梁骨间隙(图6D)。结果与图6A相似,投予个体 MLN4924可减少其胫骨因卵巢切除合并sRANKL所造成的骨质流失反应。
基于CTX-1可作为骨质再吸收及形成的生物性指标,接着利用 ELISA试剂盒来分析血清中的CTX-1含量。由图6E可知,在对小鼠投予卵巢切除术(OVX)及进行卵巢切除术后给予sRANKL(OVX+sRANKL) 皆会增加其血清中CTX-1的含量;而投予MLN4924 (OVX+sRANKL+MLN)则可回复血清中CTX-1的含量。
这些结果证实MLN4924可显著抑制骨质再吸收异常,以预防活体的骨质流失反应。
5.2生物力学特性
骨质流失会造成骨骼脆弱而易于断裂。在本实施例中,是先对小鼠投予OVX或OVX+sRANKL使其产生骨质流失后,投予MLN4924,之后利用三点弯曲试验来分析投予不同处理的小鼠股骨的生物力学特性。
如图7A-7D所述,相较于对照组,经卵巢切除术及sRANKL处理后,小鼠的包含模数(图7A)、断裂应变(图7B)、降伏压力(图7C)及断裂负载(图7D)等力学参数皆会下降;而投予MLN4924则可回复这些力学参数。换言之,MLN4924可防止因骨质流失所造成的骨骼强度/支持力度下降。
总结上述,本研究发现类泛素化抑制剂(例如MLN4924或UBA3 siRNA)的新用途,其可通过抑制破骨细胞的生成及作用来预防或治疗骨质疏松症。据此,本发明内容提供了一种用以预防或治疗骨质疏松症的方法;该方法包含对有需要的个体投予有效量的类泛素化抑制剂(例如 MLN4924或UBA3siRNA),以达到预防或治疗骨质疏松症的目的。
虽然上文实施方式中揭露了本发明的具体实施例,然其并非用以限定本发明,本领域技术人员在不悖离本发明的原理与精神的情形下,当可对其进行各种更动与修饰,因此本发明的保护范围当以附随申请专利范围所界定者为准。
序列表
<110> 长庚大学
长庚医疗财团法人嘉义长庚纪念医院
<120> 类泛素化抑制剂在制备预防或治疗骨质疏松症的药物中的用途
<130> 18FAP0090CT
<150> TW106113484
<151> 2017-04-21
<160> 8
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> RNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 1
ggagccaaug gcuguugaut t 21
<210> 2
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 2
ttccucgguu accgacaacu a 21
<210> 3
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 3
ggaucaaugg aaugcugaut t 21
<210> 4
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 4
ttccuaguua ccuuacgacu a 21
<210> 5
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 5
ccaagcucca uuguaccuut t 21
<210> 6
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 6
ttgguucgag guaacaugga a 21
<210> 7
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 7
uucuccgaac gugucacgut t 21
<210> 8
<211> 21
<212> DNA
<213> 人工序列(artificial sequence)
<220>
<221> misc_binding
<223> 合成
<400> 8
ttaagaggcu ugcacagugc a 21
Claims (5)
1.一种类泛素化(neddylation)抑制剂的用途,其是用于制备一药物以预防一个体罹患骨质疏松症或治疗一罹患骨质疏松症的个体;其中所述类泛素化抑制剂是MLN4924。
2.如权利要求1所述的用途,其中所述个体为人类,且所述类泛素化抑制剂的投予剂量为每日每公斤个体体重0.01-100毫克。
3.如权利要求2所述的用途,其中所述类泛素化抑制剂的投予剂量为每日每公斤个体体重0.1-10毫克。
4.如权利要求3所述的用途,其中所述类泛素化抑制剂的投予剂量为每日每公斤个体体重0.1-2毫克。
5.如权利要求2所述的用途,其中所述类泛素化抑制剂的投予次数为每日一次,且连续投予至少一周。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
TW106113484A TWI651087B (zh) | 2017-04-21 | 2017-04-21 | 類泛素化抑制劑於預防或治療骨質疏鬆症的用途 |
TW106113484 | 2017-04-21 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN108721627A CN108721627A (zh) | 2018-11-02 |
CN108721627B true CN108721627B (zh) | 2021-08-20 |
Family
ID=62044523
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810359098.8A Active CN108721627B (zh) | 2017-04-21 | 2018-04-20 | 类泛素化抑制剂在制备预防或治疗骨质疏松症的药物中的用途 |
Country Status (5)
Country | Link |
---|---|
US (1) | US10285994B2 (zh) |
EP (1) | EP3391880B1 (zh) |
JP (1) | JP6628832B2 (zh) |
CN (1) | CN108721627B (zh) |
TW (1) | TWI651087B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140179593A1 (en) * | 2011-05-17 | 2014-06-26 | St. Jude Children's Research Hospital | Methods and compositions for inhibiting neddylation of proteins |
CN106163557A (zh) * | 2014-01-28 | 2016-11-23 | 巴克老龄化研究所 | 用于杀死衰老细胞和用于治疗衰老相关疾病和病症的方法和组合物 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
MX2011007681A (es) * | 2009-01-19 | 2011-08-08 | Abbott Lab | Agentes inducidores de apoptosis para el tratamiento del cancer y enfermedades inmunes y autoinmunes. |
-
2017
- 2017-04-21 TW TW106113484A patent/TWI651087B/zh active
-
2018
- 2018-04-20 CN CN201810359098.8A patent/CN108721627B/zh active Active
- 2018-04-20 EP EP18168376.4A patent/EP3391880B1/en active Active
- 2018-04-20 US US15/957,951 patent/US10285994B2/en active Active
- 2018-04-20 JP JP2018081200A patent/JP6628832B2/ja active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140179593A1 (en) * | 2011-05-17 | 2014-06-26 | St. Jude Children's Research Hospital | Methods and compositions for inhibiting neddylation of proteins |
CN106163557A (zh) * | 2014-01-28 | 2016-11-23 | 巴克老龄化研究所 | 用于杀死衰老细胞和用于治疗衰老相关疾病和病症的方法和组合物 |
Also Published As
Publication number | Publication date |
---|---|
CN108721627A (zh) | 2018-11-02 |
JP6628832B2 (ja) | 2020-01-15 |
US10285994B2 (en) | 2019-05-14 |
EP3391880B1 (en) | 2023-06-07 |
JP2018184390A (ja) | 2018-11-22 |
EP3391880A1 (en) | 2018-10-24 |
TWI651087B (zh) | 2019-02-21 |
US20180303833A1 (en) | 2018-10-25 |
TW201838630A (zh) | 2018-11-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Chen et al. | Inhibition of Nrf2/HO-1 signaling leads to increased activation of the NLRP3 inflammasome in osteoarthritis | |
US11866700B2 (en) | Liposomal spherical nucleic acid (SNA) constructs presenting antisense oligonucleotides (ASO) for specific knockdown of interleukin 17 receptor mRNA | |
EP3936141A2 (en) | Anti-senescence compounds and uses thereof | |
Xiao et al. | Reduction of lactoferrin aggravates neuronal ferroptosis after intracerebral hemorrhagic stroke in hyperglycemic mice | |
Aguilar-Morante et al. | Decreased CCAAT/enhancer binding protein β expression inhibits the growth of glioblastoma cells | |
JP2021506765A (ja) | Dux4の発現に関連する疾患の治療のための化合物 | |
Shi et al. | Tangeretin suppresses osteoarthritis progression via the Nrf2/NF-κB and MAPK/NF-κB signaling pathways | |
KR101747775B1 (ko) | 유포비아 인자 l1 또는 이의 약학적으로 허용 가능한 염을 유효성분으로 함유하는 골질환의 예방 또는 치료용 약학적 조성물 | |
RU2587053C2 (ru) | Комбинированная терапия и способ оценки невосприимчивости к лечению | |
Zhou et al. | Magnolol prevents ossified tendinopathy by inhibiting PGE2-induced osteogenic differentiation of TDSCs | |
Zhao et al. | Vitamin D suppresses macrophage infiltration by down-regulation of TREM-1 in diabetic nephropathy rats | |
US20190055560A1 (en) | Fats as a target for treating tumors and uses thereof | |
Ostock et al. | Striatal norepinephrine efflux in l-DOPA-induced dyskinesia | |
US20170204151A1 (en) | Mesenchymal Stem Cells Expressing Biomarkers that Predict the Effectiveness of Mesenchymal Stem Cells for Treating Diseases and Disorders | |
CN108721627B (zh) | 类泛素化抑制剂在制备预防或治疗骨质疏松症的药物中的用途 | |
Ciociola et al. | Peyronie’s disease: a “triple oxygenant therapy” | |
CA2794581A1 (en) | Method of treating osteoporosis | |
KR101191958B1 (ko) | Tle1 억제제를 유효성분으로 함유하는 활액막 육종 예방 및 치료용 약학적 조성물 | |
JP5078124B2 (ja) | ヒト破骨細胞形成抑制活性を有する新規のペプチド | |
Zhang et al. | Fatty acid binding protein 4 (FABP4) induces chondrocyte degeneration via activation of the NF‐κb signaling pathway | |
US20160319286A1 (en) | Composition for preventing or treating osteoporosis, containing progranulin inhibitor as active ingredient | |
He et al. | Amygdalin ameliorates alopecia areata on C3H/HeJ mice by inhibiting inflammation through JAK2/STAT3 pathway | |
CN114010653A (zh) | 包括人参皂苷Rg1和岩藻多糖的组合物在制备降低组织因子的表达的药物中的应用 | |
EP3378490B1 (en) | Ectopic ossification treatment drug having mechanism for blocking par1 | |
Chen et al. | Renal lipid accumulation induced by high-fat diet regulates glucose homeostasis via sodium-glucose cotransporter 2 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |