US20210290595A1 - Gfi1 inhibitors for the treatment of hyperglycemia - Google Patents
Gfi1 inhibitors for the treatment of hyperglycemia Download PDFInfo
- Publication number
- US20210290595A1 US20210290595A1 US17/200,146 US202117200146A US2021290595A1 US 20210290595 A1 US20210290595 A1 US 20210290595A1 US 202117200146 A US202117200146 A US 202117200146A US 2021290595 A1 US2021290595 A1 US 2021290595A1
- Authority
- US
- United States
- Prior art keywords
- gfi1
- hyperglycemia
- subject
- treatment
- inhibitor
- 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.)
- Abandoned
Links
- 101150023475 Gfi1 gene Proteins 0.000 title claims abstract description 47
- 238000011282 treatment Methods 0.000 title claims abstract description 23
- 201000001421 hyperglycemia Diseases 0.000 title claims abstract description 21
- 239000003112 inhibitor Substances 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 24
- 206010012601 diabetes mellitus Diseases 0.000 claims abstract description 6
- 150000001875 compounds Chemical class 0.000 claims description 29
- 230000014509 gene expression Effects 0.000 claims description 27
- 238000012360 testing method Methods 0.000 claims description 22
- 239000003814 drug Substances 0.000 claims description 19
- 229940079593 drug Drugs 0.000 claims description 17
- 230000000694 effects Effects 0.000 claims description 17
- 108020004459 Small interfering RNA Proteins 0.000 claims description 13
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims description 9
- 239000000074 antisense oligonucleotide Substances 0.000 claims description 8
- 238000012230 antisense oligonucleotides Methods 0.000 claims description 8
- 108091034117 Oligonucleotide Proteins 0.000 claims description 4
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 claims description 4
- 150000003384 small molecules Chemical group 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 2
- 241000699670 Mus sp. Species 0.000 abstract description 27
- 241001465754 Metazoa Species 0.000 abstract description 11
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 abstract description 9
- 108020004999 messenger RNA Proteins 0.000 abstract description 9
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 abstract description 9
- 229960001052 streptozocin Drugs 0.000 abstract description 9
- 239000008194 pharmaceutical composition Substances 0.000 abstract description 6
- 230000013020 embryo development Effects 0.000 abstract description 5
- 235000009200 high fat diet Nutrition 0.000 abstract description 5
- 230000001019 normoglycemic effect Effects 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 3
- 230000003442 weekly effect Effects 0.000 abstract description 3
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 36
- 210000004027 cell Anatomy 0.000 description 27
- 239000004055 small Interfering RNA Substances 0.000 description 20
- 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 19
- 239000008103 glucose Substances 0.000 description 19
- 239000000203 mixture Substances 0.000 description 19
- 102000004877 Insulin Human genes 0.000 description 18
- 108090001061 Insulin Proteins 0.000 description 18
- 229940125396 insulin Drugs 0.000 description 18
- 210000000496 pancreas Anatomy 0.000 description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 16
- 108090000623 proteins and genes Proteins 0.000 description 16
- 239000000243 solution Substances 0.000 description 12
- 208000008589 Obesity Diseases 0.000 description 11
- 210000004369 blood Anatomy 0.000 description 11
- 239000008280 blood Substances 0.000 description 11
- 235000020824 obesity Nutrition 0.000 description 11
- 108091027967 Small hairpin RNA Proteins 0.000 description 10
- 201000010099 disease Diseases 0.000 description 10
- 239000013598 vector Substances 0.000 description 10
- 108091033409 CRISPR Proteins 0.000 description 9
- 238000012423 maintenance Methods 0.000 description 9
- 239000004382 Amylase Substances 0.000 description 8
- 102000013142 Amylases Human genes 0.000 description 8
- 108010065511 Amylases Proteins 0.000 description 8
- 108010042407 Endonucleases Proteins 0.000 description 8
- 102000004533 Endonucleases Human genes 0.000 description 8
- 235000019418 amylase Nutrition 0.000 description 8
- 238000007912 intraperitoneal administration Methods 0.000 description 8
- 150000007523 nucleic acids Chemical class 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 238000010354 CRISPR gene editing Methods 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 238000011529 RT qPCR Methods 0.000 description 6
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 description 6
- 208000035475 disorder Diseases 0.000 description 6
- 238000009472 formulation Methods 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 238000011285 therapeutic regimen Methods 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 239000004480 active ingredient Substances 0.000 description 5
- 230000015556 catabolic process Effects 0.000 description 5
- 239000006185 dispersion Substances 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- 108020000948 Antisense Oligonucleotides Proteins 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 4
- 101001059220 Homo sapiens Zinc finger protein Gfi-1 Proteins 0.000 description 4
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 description 4
- 229940100389 Sulfonylurea Drugs 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 102100029004 Zinc finger protein Gfi-1 Human genes 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 229940090124 dipeptidyl peptidase 4 (dpp-4) inhibitors for blood glucose lowering Drugs 0.000 description 4
- 230000009368 gene silencing by RNA Effects 0.000 description 4
- 238000007446 glucose tolerance test Methods 0.000 description 4
- ZNNLBTZKUZBEKO-UHFFFAOYSA-N glyburide Chemical compound COC1=CC=C(Cl)C=C1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCCCC2)C=C1 ZNNLBTZKUZBEKO-UHFFFAOYSA-N 0.000 description 4
- 238000003364 immunohistochemistry Methods 0.000 description 4
- 238000007918 intramuscular administration Methods 0.000 description 4
- 238000001990 intravenous administration Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 238000007920 subcutaneous administration Methods 0.000 description 4
- 239000003981 vehicle Substances 0.000 description 4
- 239000013585 weight reducing agent Substances 0.000 description 4
- SWLAMJPTOQZTAE-UHFFFAOYSA-N 4-[2-[(5-chloro-2-methoxybenzoyl)amino]ethyl]benzoic acid Chemical class COC1=CC=C(Cl)C=C1C(=O)NCCC1=CC=C(C(O)=O)C=C1 SWLAMJPTOQZTAE-UHFFFAOYSA-N 0.000 description 3
- 229940123208 Biguanide Drugs 0.000 description 3
- 108090000994 Catalytic RNA Proteins 0.000 description 3
- 102000053642 Catalytic RNA Human genes 0.000 description 3
- 108010051219 Cre recombinase Proteins 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 101710198884 GATA-type zinc finger protein 1 Proteins 0.000 description 3
- 102000051325 Glucagon Human genes 0.000 description 3
- 108060003199 Glucagon Proteins 0.000 description 3
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 101710163270 Nuclease Proteins 0.000 description 3
- 102100040918 Pro-glucagon Human genes 0.000 description 3
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- -1 e.g. Proteins 0.000 description 3
- 230000002124 endocrine Effects 0.000 description 3
- ZJJXGWJIGJFDTL-UHFFFAOYSA-N glipizide Chemical compound C1=NC(C)=CN=C1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCCCC2)C=C1 ZJJXGWJIGJFDTL-UHFFFAOYSA-N 0.000 description 3
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=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](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 3
- 229960004666 glucagon Drugs 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 210000004153 islets of langerhan Anatomy 0.000 description 3
- 229950004994 meglitinide Drugs 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 230000015031 pancreas development Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 108091092562 ribozyme Proteins 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- OPIFSICVWOWJMJ-AEOCFKNESA-N 5-bromo-4-chloro-3-indolyl beta-D-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CNC2=CC=C(Br)C(Cl)=C12 OPIFSICVWOWJMJ-AEOCFKNESA-N 0.000 description 2
- WOVKYSAHUYNSMH-RRKCRQDMSA-N 5-bromodeoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-RRKCRQDMSA-N 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- RKWGIWYCVPQPMF-UHFFFAOYSA-N Chloropropamide Chemical compound CCCNC(=O)NS(=O)(=O)C1=CC=C(Cl)C=C1 RKWGIWYCVPQPMF-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 101800001586 Ghrelin Proteins 0.000 description 2
- 102400000442 Ghrelin-28 Human genes 0.000 description 2
- FAEKWTJYAYMJKF-QHCPKHFHSA-N GlucoNorm Chemical compound C1=C(C(O)=O)C(OCC)=CC(CC(=O)N[C@@H](CC(C)C)C=2C(=CC=CC=2)N2CCCCC2)=C1 FAEKWTJYAYMJKF-QHCPKHFHSA-N 0.000 description 2
- 208000013016 Hypoglycemia Diseases 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 2
- 206010022489 Insulin Resistance Diseases 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 102000000574 RNA-Induced Silencing Complex Human genes 0.000 description 2
- 108010016790 RNA-Induced Silencing Complex Proteins 0.000 description 2
- 102000005157 Somatostatin Human genes 0.000 description 2
- 108010056088 Somatostatin Proteins 0.000 description 2
- 238000010459 TALEN Methods 0.000 description 2
- 229940123464 Thiazolidinedione Drugs 0.000 description 2
- 108010043645 Transcription Activator-Like Effector Nucleases Proteins 0.000 description 2
- 108010017070 Zinc Finger Nucleases Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 102000005936 beta-Galactosidase Human genes 0.000 description 2
- 108010005774 beta-Galactosidase Proteins 0.000 description 2
- 150000004283 biguanides Chemical class 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 231100000504 carcinogenesis Toxicity 0.000 description 2
- 230000030570 cellular localization Effects 0.000 description 2
- OSASVXMJTNOKOY-UHFFFAOYSA-N chlorobutanol Chemical compound CC(C)(O)C(Cl)(Cl)Cl OSASVXMJTNOKOY-UHFFFAOYSA-N 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 102000038379 digestive enzymes Human genes 0.000 description 2
- 108091007734 digestive enzymes Proteins 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000002612 dispersion medium Substances 0.000 description 2
- 230000034431 double-strand break repair via homologous recombination Effects 0.000 description 2
- 229940029980 drug used in diabetes Drugs 0.000 description 2
- 210000001198 duodenum Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- BGHSOEHUOOAYMY-JTZMCQEISA-N ghrelin Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)CN)C1=CC=CC=C1 BGHSOEHUOOAYMY-JTZMCQEISA-N 0.000 description 2
- WIGIZIANZCJQQY-RUCARUNLSA-N glimepiride Chemical compound O=C1C(CC)=C(C)CN1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)N[C@@H]2CC[C@@H](C)CC2)C=C1 WIGIZIANZCJQQY-RUCARUNLSA-N 0.000 description 2
- 230000002641 glycemic effect Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000013632 homeostatic process Effects 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 230000001771 impaired effect Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000000415 inactivating effect Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 230000009707 neogenesis Effects 0.000 description 2
- 230000006780 non-homologous end joining Effects 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 2
- QGJUIPDUBHWZPV-SGTAVMJGSA-N saxagliptin Chemical compound C1C(C2)CC(C3)CC2(O)CC13[C@H](N)C(=O)N1[C@H](C#N)C[C@@H]2C[C@@H]21 QGJUIPDUBHWZPV-SGTAVMJGSA-N 0.000 description 2
- 108010033693 saxagliptin Proteins 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 2
- 229960000553 somatostatin Drugs 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 150000001467 thiazolidinediones Chemical class 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000011830 transgenic mouse model Methods 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- 241001430294 unidentified retrovirus Species 0.000 description 2
- OELFLUMRDSZNSF-OFLPRAFFSA-N (2R)-2-[[oxo-(4-propan-2-ylcyclohexyl)methyl]amino]-3-phenylpropanoic acid Chemical compound C1CC(C(C)C)CCC1C(=O)N[C@@H](C(O)=O)CC1=CC=CC=C1 OELFLUMRDSZNSF-OFLPRAFFSA-N 0.000 description 1
- GQPYTJVDPQTBQC-KLQYNRQASA-N (3r)-3-amino-1-[3-(trifluoromethyl)-6,8-dihydro-5h-[1,2,4]triazolo[4,3-a]pyrazin-7-yl]-4-(2,4,5-trifluorophenyl)butan-1-one;phosphoric acid;hydrate Chemical compound O.OP(O)(O)=O.C([C@H](CC(=O)N1CC=2N(C(=NN=2)C(F)(F)F)CC1)N)C1=CC(F)=C(F)C=C1F GQPYTJVDPQTBQC-KLQYNRQASA-N 0.000 description 1
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- 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
- 229940077274 Alpha glucosidase inhibitor Drugs 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 244000303258 Annona diversifolia Species 0.000 description 1
- 235000002198 Annona diversifolia Nutrition 0.000 description 1
- 108020004491 Antisense DNA Proteins 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 108091023037 Aptamer Proteins 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- XNCOSPRUTUOJCJ-UHFFFAOYSA-N Biguanide Chemical compound NC(N)=NC(N)=N XNCOSPRUTUOJCJ-UHFFFAOYSA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- CZKGBIYJXUXXNP-ZWKOTPCHSA-N CN1C=C(C(=O)CC2=CC=C([C@@H]3C[C@H]3NCC3CC3)C=C2)C=N1.Cl Chemical compound CN1C=C(C(=O)CC2=CC=C([C@@H]3C[C@H]3NCC3CC3)C=C2)C=N1.Cl CZKGBIYJXUXXNP-ZWKOTPCHSA-N 0.000 description 1
- NTRMVYTYDOLCRU-DLBZAZTESA-N C[n]1ncc(C(Nc2ccc([C@H](C3)[C@@H]3NCC3CC3)cc2)=O)c1 Chemical compound C[n]1ncc(C(Nc2ccc([C@H](C3)[C@@H]3NCC3CC3)cc2)=O)c1 NTRMVYTYDOLCRU-DLBZAZTESA-N 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 230000033616 DNA repair Effects 0.000 description 1
- 241000252212 Danio rerio Species 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000991587 Enterovirus C Species 0.000 description 1
- 241000702191 Escherichia virus P1 Species 0.000 description 1
- 241000589601 Francisella Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 108020005004 Guide RNA Proteins 0.000 description 1
- 241000713858 Harvey murine sarcoma virus Species 0.000 description 1
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- 206010060378 Hyperinsulinaemia Diseases 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- 101710147301 MOG interacting and ectopic P-granules protein 1 Proteins 0.000 description 1
- 101710175625 Maltose/maltodextrin-binding periplasmic protein Proteins 0.000 description 1
- 102100025169 Max-binding protein MNT Human genes 0.000 description 1
- 241000713869 Moloney murine leukemia virus Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241001631646 Papillomaviridae Species 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241001505332 Polyomavirus sp. Species 0.000 description 1
- 101710183548 Pyridoxal 5'-phosphate synthase subunit PdxS Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- YASAKCUCGLMORW-UHFFFAOYSA-N Rosiglitazone Chemical compound C=1C=CC=NC=1N(C)CCOC(C=C1)=CC=C1CC1SC(=O)NC1=O YASAKCUCGLMORW-UHFFFAOYSA-N 0.000 description 1
- 241000714474 Rous sarcoma virus Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- 241000193996 Streptococcus pyogenes Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- 241000269457 Xenopus tropicalis Species 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000002679 ablation Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 210000005006 adaptive immune system Anatomy 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 239000003888 alpha glucosidase inhibitor Substances 0.000 description 1
- 229940000806 amaryl Drugs 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 239000003816 antisense DNA Substances 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 229920002988 biodegradable polymer Polymers 0.000 description 1
- 239000004621 biodegradable polymer Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000001815 biotherapy Methods 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000007248 cellular mechanism Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229960004926 chlorobutanol Drugs 0.000 description 1
- 229960001761 chlorpropamide Drugs 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 238000011340 continuous therapy Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- WZHCOOQXZCIUNC-UHFFFAOYSA-N cyclandelate Chemical compound C1C(C)(C)CC(C)CC1OC(=O)C(O)C1=CC=CC=C1 WZHCOOQXZCIUNC-UHFFFAOYSA-N 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 229940089126 diabeta Drugs 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- UGMCXQCYOVCMTB-UHFFFAOYSA-K dihydroxy(stearato)aluminium Chemical compound CCCCCCCCCCCCCCCCCC(=O)O[Al](O)O UGMCXQCYOVCMTB-UHFFFAOYSA-K 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000005782 double-strand break Effects 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 230000004049 epigenetic modification Effects 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000012458 free base Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 238000012226 gene silencing method Methods 0.000 description 1
- 230000004077 genetic alteration Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010362 genome editing Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 229960004580 glibenclamide Drugs 0.000 description 1
- 229960004346 glimepiride Drugs 0.000 description 1
- 229960001381 glipizide Drugs 0.000 description 1
- 229940095884 glucophage Drugs 0.000 description 1
- 230000009229 glucose formation Effects 0.000 description 1
- 230000014101 glucose homeostasis Effects 0.000 description 1
- 229940088991 glucotrol Drugs 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 229940120105 glynase Drugs 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 208000016354 hearing loss disease Diseases 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 235000011167 hydrochloric acid Nutrition 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 230000003451 hyperinsulinaemic effect Effects 0.000 description 1
- 201000008980 hyperinsulinism Diseases 0.000 description 1
- 230000003463 hyperproliferative effect Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000005462 in vivo assay Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 239000007972 injectable composition Substances 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000010189 intracellular transport Effects 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 229940090473 januvia Drugs 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229960003105 metformin Drugs 0.000 description 1
- OETHQSJEHLVLGH-UHFFFAOYSA-N metformin hydrochloride Chemical compound Cl.CN(C)C(=N)N=C(N)N OETHQSJEHLVLGH-UHFFFAOYSA-N 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000003990 molecular pathway Effects 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229940001450 onglyza Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000004203 pancreatic function Effects 0.000 description 1
- 210000004923 pancreatic tissue Anatomy 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 239000000816 peptidomimetic Substances 0.000 description 1
- 229960003742 phenol Drugs 0.000 description 1
- 150000004713 phosphodiesters Chemical class 0.000 description 1
- 235000011007 phosphoric acid Nutrition 0.000 description 1
- 150000003016 phosphoric acids Chemical class 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- HYAFETHFCAUJAY-UHFFFAOYSA-N pioglitazone Chemical compound N1=CC(CC)=CC=C1CCOC(C=C1)=CC=C1CC1C(=O)NC(=O)S1 HYAFETHFCAUJAY-UHFFFAOYSA-N 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 230000007542 postnatal development Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M potassium chloride Inorganic materials [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 1
- 229940096058 prandin Drugs 0.000 description 1
- 230000009237 prenatal development Effects 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- MFDFERRIHVXMIY-UHFFFAOYSA-N procaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 MFDFERRIHVXMIY-UHFFFAOYSA-N 0.000 description 1
- 229960004919 procaine Drugs 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 108020001580 protein domains Proteins 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 description 1
- 229960002354 repaglinide Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 229960004937 saxagliptin Drugs 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 239000002924 silencing RNA Substances 0.000 description 1
- 230000005783 single-strand break Effects 0.000 description 1
- 229960004034 sitagliptin Drugs 0.000 description 1
- MFFMDFFZMYYVKS-SECBINFHSA-N sitagliptin Chemical compound C([C@H](CC(=O)N1CC=2N(C(=NN=2)C(F)(F)F)CC1)N)C1=CC(F)=C(F)C=C1F MFFMDFFZMYYVKS-SECBINFHSA-N 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- YROXIXLRRCOBKF-UHFFFAOYSA-N sulfonylurea Chemical class OC(=N)N=S(=O)=O YROXIXLRRCOBKF-UHFFFAOYSA-N 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 108091006107 transcriptional repressors Proteins 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001529453 unidentified herpesvirus Species 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000012800 visualization Methods 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/415—1,2-Diazoles
-
- 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
-
- 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
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- 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
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
-
- 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.
Definitions
- the present invention relates to methods and pharmaceutical compositions useful for the treatment of hyperglycemia.
- the pancreas is an abdominal gland located behind the stomach and connected to the duodenum. It is divided into two functionally and morphologically distinct compartments: the exocrine and the endocrine tissues.
- the exocrine compartment (encompassing 98% of the total organ mass) represents a “factory” producing digestive enzymes that catalyze the breakdown of proteins, carbohydrates, and lipids.
- Pancreatic digestive enzymes are collected by intercalated ducts and are subsequently emptied into the duodenum (1).
- the endocrine compartment (representing less than 2% of the pancreatic tissue) is organized into highly vascularized and innervated cell clusters termed islets of Langerhans.
- Glucagon has the opposite function by promoting glucose release and/or neogenesis from glycogen, in case of hypoglycemia.
- the present invention relates to methods and pharmaceutical compositions useful for the treatment of hyperglycemia.
- the present invention is defined by the claims.
- the first object of the present relates to a method of treating hyperglycemia in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an inhibitor of Gfi1.
- hyperglycemia has its general meaning in the art and refers to any elevated level of blood glucose compared to a basal level in a subject.
- Basal level refers to a blood glucose level of a normal subject when fasting.
- hyperglycemia refers to blood glucose levels above about 100 mg/dl.
- hyperglycemia refers to blood glucose levels above about 110 mg/dl when fasting and above about 140 mg/dl two hours after having a meal.
- the inhibitor of Gfi1 is thus particularly suitable for reducing blood glucose levels and thus for achieving normoglycemia in the subject.
- the expression “achieving normoglycemia” refers to the maintenance of blood glucose concentrations at the basal level.
- the subject suffers from diabetes.
- diabetes has its general meaning in the art and refers to the chronic disease characterized by relative or absolute deficiency of insulin that results in hyperglycemia.
- the term “diabetes” is thus intended to include those individuals with hyperglycemia, including chronic hyperglycemia, hyperinsulinemia, impaired glucose homeostasis or tolerance, and insulin resistance.
- the subject suffers from Type 1 diabetes mellitus.
- Type 1 diabetes mellitus or “T1D” has its general meaning in the art and refers to an autoimmune disorder than leads to destruction of the insulin producing beta cells of the pancreas leading to hyperglycemia.
- the subject suffers from type 2 diabetes.
- type 2 diabetes or “non-insulin dependent diabetes mellitus (NIDDM)” has its general meaning in the art. Type 2 diabetes often occurs when levels of insulin are normal or even elevated and appears to result from the inability of tissues to respond appropriately to insulin. Most of the type 2 diabetics are obese.
- Obesity refers to a condition characterized by an excess of body fat. The operational definition of obesity is based on the Body Mass Index (BMI), which is calculated as body weight per height in meter squared (kg/m 2 ).
- BMI Body Mass Index
- Obesity refers to a condition whereby an otherwise healthy subject has a BMI greater than or equal to 30 kg/m 2 , or a condition whereby a subject with at least one co-morbidity has a BMI greater than or equal to 27 kg/m 2 .
- An “obese subject” is an otherwise healthy subject with a BMI greater than or equal to 30 kg/m 2 or a subject with at least one co-morbidity with a BMI greater than or equal 27 kg/m 2 .
- a “subject at risk of obesity” is an otherwise healthy subject with a BMI of 25 kg/m 2 to less than 30 kg/m 2 or a subject with at least one co-morbidity with a BMI of 25 kg/m 2 to less than 27 kg/m 2 .
- the increased risks associated with obesity may occur at a lower BMI in people of Asian descent.
- “obesity” refers to a condition whereby a subject with at least one obesity-induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, has a BMI greater than or equal to 25 kg/m 2 .
- An “obese subject” in these countries refers to a subject with at least one obesity-induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, with a BMI greater than or equal to 25 kg/m 2 .
- a “subject at risk of obesity” is a person with a BMI of greater than 23 kg/m 2 to less than 25 kg/m 2 .
- treatment refers to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse.
- the treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.
- therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
- a therapeutic regimen may include an induction regimen and a maintenance regimen.
- the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
- the general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen.
- An induction regimen may employ (in part or in whole) a “loading regimen”, which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
- maintenance regimen refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years).
- a maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).
- Gfi1 has its general meaning in the art and refers to the growth factor independent 1 transcriptional repressor (gene ID 2672). Gfi1 is also known as SCN2; GFI-1; GFI1A; ZNF163. This gene encodes a nuclear zinc finger protein that functions as a transcriptional repressor. This protein plays a role in diverse developmental contexts, including hematopoiesis and oncogenesis. Exemplary human nucleic acid and amino acid sequences are represented by the NCBI reference sequences NM_001127215.1 and NP_001120687.1 respectively.
- an “inhibitor of Gfi1” refers to a compound, substance or composition that can inhibit the function of Gfi1.
- the inhibitor can inhibit the expression or activity of Gfi1.
- Gfi1 inhibitors include but are not limited to polypeptides such as dominant-negative protein mutants, peptidomimetics, antibodies, ribozymes, antisense oligonucleotides, or other small organic molecules which specifically inhibit the activity or expression of Gfi1.
- small organic molecule refers to a molecule of a size comparable to those organic molecules generally used in pharmaceuticals.
- Preferred small organic molecules range in size up to about 5000 Da, more preferably up to 2000 Da, and most preferably up to about 1000 Da.
- the small molecule is T-3775440 as described in Ishikawa et al 2016. This molecule has the following structure in the art:
- the inhibitor is a short hairpin RNA (shRNA), a small interfering RNA (siRNA) or an antisense oligonucleotide which inhibits the expression of Gfi1.
- shRNA short hairpin RNA
- shRNA is a sequence of RNA that makes a tight hairpin turn that can be used to silence gene expression via RNA interference.
- shRNA is generally expressed using a vector introduced into cells, wherein the vector utilizes the U6 promoter to ensure that the shRNA is always expressed. This vector is usually passed on to daughter cells, allowing the gene silencing to be inherited.
- the shRNA hairpin structure is cleaved by the cellular machinery into siRNA, which is then bound to the RNA-induced silencing complex (RISC).
- RISC RNA-induced silencing complex
- siRNA Small interfering RNA
- silencing RNA are a class of 20-25 nucleotide-long double-stranded RNA molecules that play a variety of roles in biology. Most notably, siRNA is involved in the RNA interference (RNAi) pathway whereby the siRNA interferes with the expression of a specific gene.
- RNAi RNA interference
- Anti-sense oligonucleotides include anti-sense RNA molecules and anti-sense DNA molecules, would act to directly block the translation of the targeted mRNA by binding thereto and thus preventing protein translation or increasing mRNA degradation, thus decreasing the level of the targeted protein, and thus activity, in a cell.
- antisense oligonucleotides of at least about 15 bases and complementary to unique regions of the mRNA transcript sequence can be synthesized, e.g., by conventional phosphodiester techniques. Methods for using antisense techniques for specifically inhibiting gene expression of genes whose sequence is known are well known in the art (e.g. see U.S. Pat. Nos.
- Antisense oligonucleotides, siRNAs, shRNAs of the invention may be delivered in vivo alone or in association with a vector.
- a “vector” is any vehicle capable of facilitating the transfer of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid to the cells and typically mast cells.
- the vector transports the nucleic acid to cells with reduced degradation relative to the extent of degradation that would result in the absence of the vector.
- the vectors useful in the invention include, but are not limited to, plasmids, phagemids, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid sequences.
- Viral vectors are a preferred type of vector and include, but are not limited to nucleic acid sequences from the following viruses: retrovirus, such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma virus; adenovirus, adeno-associated virus; SV40-type viruses; polyoma viruses; Epstein-Barr viruses; papilloma viruses; herpes virus; vaccinia virus; polio virus; and RNA virus such as a retrovirus.
- retrovirus such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma virus
- adenovirus adeno-associated virus
- SV40-type viruses polyoma viruses
- Epstein-Barr viruses Epstein-Barr viruses
- papilloma viruses herpes virus
- vaccinia virus
- the inhibitor of Gif1 expression is an endonuclease.
- sequencing technologies have provided an unprecedentedly detailed overview of the multiple genetic aberrations in cancer.
- these new data strongly emphasize the need of fast and reliable strategies to characterize the normal and pathological function of these genes and assess their role, in particular as driving factors during oncogenesis.
- the new technologies provide the means to recreate the actual mutations observed in cancer through direct manipulation of the genome. Indeed, natural and engineered nuclease enzymes have attracted considerable attention in the recent years.
- Endonuclease-based genome inactivating generally requires a first step of DNA single or double strand break, which can then trigger two distinct cellular mechanisms for DNA repair, which can be exploited for DNA inactivating: the errorprone nonhomologous end-joining (NHEJ) and the high-fidelity homology-directed repair (HDR).
- Endonucleases for gene inactivation have come in various forms, which includes CRISPR)/CRISPR associated (Cas) systems, mega nucleases (MN), zinc finger nucleases (ZFN), and transcription activator-like effector nucleases (TALEN).
- Endonucleases for use in the present invention are disclosed in WO 2010/079430, WO2011072246, WO2013045480, Mussolino C, et al (Curr Opin Biotechnol. 2012 October; 23(5):644-50) and Papaioannou I. et al (Expert Opinion on Biological Therapy, March 2012, Vol. 12, No. 3: 329-342).
- the endonuclease is CRISPR-cas.
- CRISPR-cas has its general meaning in the art and refers to clustered regularly interspaced short palindromic repeats associated which are the segments of prokaryotic DNA containing short repetitions of base sequences.
- the endonuclease is CRISPR-cas9 which is from Streptococcus pyogenes .
- the CRISPR/Cas9 system has been described in U.S. Pat. No. 8,697,359 B1 and US 2014/0068797.
- CRISPR has been recently engineered into a new powerful tool for genome editing. It has already been successfully used to target important genes in many cell lines and organisms, including human (Mali et al., 2013, Science, Vol. 339: 823-826), bacteria (Fabre et al., 2014, PLoS Negl. Trop. Dis., Vol.
- the endonuclease is CRISPR-Cpf1 which is the more recently characterized CRISPR from Provotella and Francisella 1 (Cpf1) in Zetsche et al. (“Cpf1 is a Single RNA-guided Endonuclease of a Class 2 CRISPR-Cas System (2015); Cell; 163, 1-13).
- the inhibitor is an intrabody having specificity for Gfi1.
- the term “intrabody” generally refer to an intracellular antibody or antibody fragment.
- Antibodies in particular single chain variable antibody fragments (scFv), can be modified for intracellular localization. Such modification may entail for example, the fusion to a stable intracellular protein, such as, e.g., maltose binding protein, or the addition of intracellular trafficking/localization peptide sequences, such as, e.g., the endoplasmic reticulum retention.
- the intrabody is a single domain antibody.
- the antibody according to the invention is a single domain antibody.
- single domain antibody sdAb or “VHH” refers to the single heavy chain variable domain of antibodies of the type that can be found in Camelid mammals which are naturally devoid of light chains. Such VHH are also called “Nanobody®”. According to the invention, sdAb can particularly be llama sdAb.
- administering refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., an inhibitor of Gif1) into the subject, such as by mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery and/or any other method of physical delivery described herein or known in the art.
- a disease, or a symptom thereof is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof.
- administration of the substance typically occurs before the onset of the disease or symptoms thereof.
- a “therapeutically effective amount” of the inhibitor as above described is meant a sufficient amount to provide a therapeutic effect. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment.
- the specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific polypeptide employed; and like factors well known in the medical arts.
- the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
- the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated.
- a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient.
- An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day.
- the inhibitor is administered to the subject in the form of a pharmaceutical composition.
- the inhibitor may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions.
- pharmaceutically acceptable excipients such as a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a carboxylate, a pharmaceutically acceptable.
- pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.
- the active principle in the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings.
- Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.
- the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected.
- vehicles which are pharmaceutically acceptable for a formulation capable of being injected.
- These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions.
- the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.
- the form In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.
- Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
- the inhibitor can be formulated into a composition in a neutral or salt form.
- Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.
- inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like.
- Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine,
- the carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
- the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
- the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
- isotonic agents for example, sugars or sodium chloride.
- Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.
- Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization.
- dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above.
- sterile powders for the preparation of sterile injectable solutions the typical methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
- the preparation of more, or highly concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents.
- solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective.
- the formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed.
- aqueous solutions For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose.
- aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.
- sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
- the inhibitor of the present invention is administered to the subject in combination with another diabetes medications.
- exemplary diabetes medications include sulfonylureas, meglitinides, biguanides, thiazolidinediones, alpha-glucosidase inhibitors, or DPP-4 inhibitors.
- Sulfonylureas stimulate the beta cells of the pancreas to release more insulin.
- Chlorpropamide (Diabinese) is the only first-generation sulfonylurea still in use today. The second generation sulfonylureas are used in smaller doses than the first-generation drugs.
- glipizide Glucotrol and Glucotrol XL
- glyburide Micronase, Glynase, and Diabeta
- glimepiride Amaryl
- Meglitinides are drugs that also stimulate the beta cells to release insulin.
- Repaglinide Prandin
- nateglinide Starlix
- Metformin Glucophage
- Biguanides lower blood glucose levels primarily by decreasing the amount of glucose produced by the liver.
- Rosiglitazone Avandia
- pioglitazone ACTOS
- DPP-4 inhibitors help improve AIC without causing hypoglycemia. They work by preventing the breakdown of a naturally occurring compound in the body, GLP-1. GLP-1 reduces blood glucose levels in the body, but is broken down very quickly so it does not work well when injected as a drug itself. By interfering in the process that breaks down GLP-1, DPP-4 inhibitors allow it to remain active in the body longer, lowering blood glucose levels only when they are elevated.
- Sitagliptin JANUVIA
- saxagliptin ONGLYZA
- a further object of the present invention relates to a method of screening a drug suitable for the treatment of hyperglycemia comprising i) providing a test compound and ii) determining the ability of said test compound to inhibit the expression or activity of Gfi1.
- the assay first comprises determining the ability of the test compound to bind to Gfi1.
- a population of pancreatic cells (acinar, ductal or endocrine) is then contacted and activated so as to determine the ability of the test compound to inhibit the activity or expression of Gfi1.
- the effect triggered by the test compound is determined relative to that of a population of immune cells incubated in parallel in the absence of the test compound or in the presence of a control agent either of which is analogous to a negative control condition.
- control substance refers a molecule that is inert or has no activity relating to an ability to modulate a biological activity or expression. It is to be understood that test compounds capable of inhibiting the activity or expression of Gfi1, as determined using in vitro methods described herein, are likely to exhibit similar modulatory capacity in applications in vivo. In vivo assays are well known in the art and typically include those described in the EXAMPLE. Typically, the test compound is selected from the group consisting of peptides, petptidomimetics, small organic molecules, antibodies (e.g. intraantibodies), aptamers or nucleic acids.
- test compound according to the invention may be selected from a library of compounds previously synthesised, or a library of compounds for which the structure is determined in a database, or from a library of compounds that have been synthesised de novo.
- the test compound may be selected form small organic molecules.
- FIG. 1 qRT-PCR quantification of Gfi1 expression in mouse pancreas from E15.5 (embryonic day 15.5) to 11.8 months of age.
- FIG. 2 X-gal staining and immunohistochemistry of Gfi1Cre::Rosa-lox--stop-lox-beta-Gal pancreas.
- FIG. 3 Immunohistochemistry of adult pancreas labelled with insulin, amylase and DAPI.
- FIG. 4 qRT-PCR assessment of amylase gene expression from whole pancreas at different developmental and adult stages.
- FIG. 5 Immunohistochemical assessment of BrdU-labelled cells.
- FIG. 6 Quantitative analyses of BrdU-labelled cells in different pancreatic compartments
- FIG. 7A-D Intraperitoneal glucose tolerance test on 3 months old mice (A) and 6.5 months old mice (B). Intraperitoneal insulin tolerance test on 3 months old mice (C) and 6.5 months old mice (D).
- FIG. 8A-D Weight and basal glycemia monitoring of control and Gfi1 mutant mice subjected to high fat diet (A-B). Glucose tolerance test (C) and insulin tolerance test (D) performed on mice of the indicated genotypes following exposure to high fat diet for 5 months.
- FIG. 9 Basal glycaemia monitoring of control and Gfi1 mutant mice treated with high dose streptozotocin. Arrow represent days of streptozotocin injection.
- FIG. 10 Insulin immunostaining of pancreatic sections 5 days after acute streptozotocin treatment.
- FIG. 11 qRT-PCR quantification of NKX6.2 mRNA levels from whole pancreas at different developmental and adult stages.
- FIG. 12 RNA scope targeting NKX6.2 mRNA
- Gfi1 is expressed in the pancreas, starting from the first stages of pancreatic embryonic development. Furthermore, we observed that Gfi1 mRNA levels remain steady throughout embryonic development, while they significantly increase during the first days of life (from Postnatal day 6). A further increase was also noted between 1 and 3 months of age. Afterwards Gfi1 expression appeared to be stable during adulthood until at least 12 months of life ( FIG. 1 ).
- Gfi1-Cre mouse line expressing the phage P1 Cre Recombinase under the control of the endogenous Gfi1 promoter
- a ROSA26- ⁇ -gal mouse line harvested from a transgene containing the Rosa26 promoter upstream of a loxP-Neomycin resistance-Stop-loxP- ⁇ -galactosidase cassette.
- cells are permanently labelled once Gfi1 expression is activated.
- the spatiotemporal expression of ⁇ -galactosidase driven by Gfi1 promoter was analyzed by X-gal staining.
- ⁇ -galactosidase activity was solely detected in the acinar compartment. Importantly, no activity was found in ductal nor islets of Langerhans cells, demonstrating that the latter do not derive from cells having expressed Gfi1 ( FIG. 2 ).
- Gfi1 null animals suffer from several health conditions: loss of hearing, a general growth deficiency and severe neutropenia that impairs their life expectancy to a maximum of 1 month.
- loss of hearing a general growth deficiency and severe neutropenia that impairs their life expectancy to a maximum of 1 month.
- a conditional knock out system where Gfi1 expression is silenced exclusively in the pancreas from the first phases of development.
- Pdx1Cre::Gfi1cko transgenic mouse line (16) In the resulting animals, the Pdx1 promoter drives the expression of the Cre recombinase in all pancreatic cells.
- Pdx1Cre::Gfi1cko pancreata were thoroughly analyzed by immunohistochemistry. Our phenotypical characterization showed that the majority of acinar cells were either negative or weakly positive for amylase ( FIG. 3 ). To confirm these observations, the amount of amylase transcript was assessed by RT-qPCR at different developmental and adult (st)ages. As expected, amylase expression levels increased significantly from E15.5 throughout postnatal life of control animals up to 3 months of age. Importantly, amylase transcript contents also raised continuously throughout pre- and post-natal development, but these were consistently decreased compared to controls at all ages tested. Strikingly, a significant 78.3% decrease in amylase expression level was measured in 3 months old Gfi1-mutants ( FIG. 4 ).
- Gfi1 mutant acinar cells are hyperproliferative. Indeed, following 5 days of BrdU supplementation, BrdU immunoreactive cells were rarely observed in 3 months old control pancreata while an impressive number of dividing cells was detected in pancreatic sections isolated from adult Gfi1-deficient mice ( FIG. 5 ). Quantitative analyses demonstrated that the increase in the cellular proliferation rate was restricted to the acinar compartment ( FIG. 6 ).
- Pdx1Cre::Gfi1cko mice were significantly more responsive to exogenous insulin administration compared to age matched controls ( FIG. 7D ).
- mutant mice remained normoglycemic (with a glycemia lower than 120 mg/dl) throughout the entire experiment ( FIG. 8B ).
- mice were sacrificed 5 days after streptozotocin injection and pancreas specimens were analyzed by immunohistochemistry. Surprisingly, the ⁇ -cell mass and islets of Langerhans integrity were found unaffected by streptozotocin cytotoxic activity in Gfi1 conditional knockout pancreata ( FIG. 10 ).
- pancreatic transcriptome extracted from Gfi1-deficient mice by RT-qPCR and assessed the expression level of all genes known for their involvement in pancreas development and function.
- Nkx6.2 was abnormally expressed in Pdx1Cre::Gfi1cko mice.
- Nkx6.2 transcripts were solely detected at E15.5 in control pancreas, Nkx6.2 being subsequently switched off ( FIG. 11 ).
- Pdx1Cre::Gfi1cko pancreatic cells failed to inactivate Nkx6.2 expression during the latest stages of embryonic development ( FIG. 11 ).
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Diabetes (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Emergency Medicine (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Endocrinology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
- The present invention relates to methods and pharmaceutical compositions useful for the treatment of hyperglycemia.
- The pancreas is an abdominal gland located behind the stomach and connected to the duodenum. It is divided into two functionally and morphologically distinct compartments: the exocrine and the endocrine tissues. The exocrine compartment (encompassing 98% of the total organ mass) represents a “factory” producing digestive enzymes that catalyze the breakdown of proteins, carbohydrates, and lipids. Pancreatic digestive enzymes are collected by intercalated ducts and are subsequently emptied into the duodenum (1). The endocrine compartment (representing less than 2% of the pancreatic tissue) is organized into highly vascularized and innervated cell clusters termed islets of Langerhans. These comprise five different hormone-secreting cell subtypes: α-, β-, δ-, ε-, and PP-cells secreting respectively glucagon, insulin, somatostatin, ghrelin, and PP (pancreatic polypeptide—1-3). Insulin and glucagon act coordinately to maintain glycemic homeostasis by regulating the storage, metabolism, and neo-genesis of glucose. Insulin is released in response to increased blood glucose levels and acts by reducing glycemia through the induction of the storage and metabolism of glucose. Glucagon has the opposite function by promoting glucose release and/or neogenesis from glycogen, in case of hypoglycemia. Somatostatin and PP have been implicated in the regulation of other hormones and exocrine enzyme secretion (4-6). The function of ghrelin, synthesized and secreted by ε-cells, is still unclear regarding glycemic homeostasis (7-9). Classical genetic approaches have revealed much about individual factors regulating pancreatic development (10), however, we have yet to understand the regulatory network underlying pancreas formation (11) and all the factors involved in this process. Some of these factors are well known and studied but increasing researches revealed new and unknown factors involved in pancreas development and maturation.
- The present invention relates to methods and pharmaceutical compositions useful for the treatment of hyperglycemia. In particular, the present invention is defined by the claims.
- The first object of the present relates to a method of treating hyperglycemia in a subject in need thereof comprising administering to the subject a therapeutically effective amount of an inhibitor of Gfi1.
- As used herein, the term “hyperglycemia has its general meaning in the art and refers to any elevated level of blood glucose compared to a basal level in a subject. “Basal level”, as used herein, refers to a blood glucose level of a normal subject when fasting. Generally, hyperglycemia refers to blood glucose levels above about 100 mg/dl. In particular, hyperglycemia refers to blood glucose levels above about 110 mg/dl when fasting and above about 140 mg/dl two hours after having a meal.
- According to the present invention, the inhibitor of Gfi1 is thus particularly suitable for reducing blood glucose levels and thus for achieving normoglycemia in the subject. As used herein, the expression “achieving normoglycemia” refers to the maintenance of blood glucose concentrations at the basal level.
- In some embodiments, the subject suffers from diabetes. As used herein, the term “diabetes” has its general meaning in the art and refers to the chronic disease characterized by relative or absolute deficiency of insulin that results in hyperglycemia. The term “diabetes” is thus intended to include those individuals with hyperglycemia, including chronic hyperglycemia, hyperinsulinemia, impaired glucose homeostasis or tolerance, and insulin resistance.
- In some embodiments, the subject suffers from
Type 1 diabetes mellitus. As used herein, the term “Type 1 diabetes mellitus” or “T1D” has its general meaning in the art and refers to an autoimmune disorder than leads to destruction of the insulin producing beta cells of the pancreas leading to hyperglycemia. - In some embodiments, the subject suffers from
type 2 diabetes. As used herein, the term “type 2 diabetes” or “non-insulin dependent diabetes mellitus (NIDDM)” has its general meaning in the art.Type 2 diabetes often occurs when levels of insulin are normal or even elevated and appears to result from the inability of tissues to respond appropriately to insulin. Most of thetype 2 diabetics are obese. As used herein the term “obesity” refers to a condition characterized by an excess of body fat. The operational definition of obesity is based on the Body Mass Index (BMI), which is calculated as body weight per height in meter squared (kg/m2). Obesity refers to a condition whereby an otherwise healthy subject has a BMI greater than or equal to 30 kg/m2, or a condition whereby a subject with at least one co-morbidity has a BMI greater than or equal to 27 kg/m2. An “obese subject” is an otherwise healthy subject with a BMI greater than or equal to 30 kg/m2 or a subject with at least one co-morbidity with a BMI greater than or equal 27 kg/m2. A “subject at risk of obesity” is an otherwise healthy subject with a BMI of 25 kg/m2 to less than 30 kg/m2 or a subject with at least one co-morbidity with a BMI of 25 kg/m2 to less than 27 kg/m2. The increased risks associated with obesity may occur at a lower BMI in people of Asian descent. In Asian and Asian-Pacific countries, including Japan, “obesity” refers to a condition whereby a subject with at least one obesity-induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, has a BMI greater than or equal to 25 kg/m2. An “obese subject” in these countries refers to a subject with at least one obesity-induced or obesity-related co-morbidity that requires weight reduction or that would be improved by weight reduction, with a BMI greater than or equal to 25 kg/m2. In these countries, a “subject at risk of obesity” is a person with a BMI of greater than 23 kg/m2 to less than 25 kg/m2. - As used herein, the term “treatment” or “treat” refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. By “therapeutic regimen” is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy. A therapeutic regimen may include an induction regimen and a maintenance regimen. The phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease. The general goal of an induction regimen is to provide a high level of drug to a patient during the initial period of a treatment regimen. An induction regimen may employ (in part or in whole) a “loading regimen”, which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both. The phrase “maintenance regimen” or “maintenance period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a patient during treatment of an illness, e.g., to keep the patient in remission for long periods of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering a drug at a regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of a particular predetermined criteria [e.g., disease manifestation, etc.]).
- As used herein, the term “Gfi1” has its general meaning in the art and refers to the growth factor independent 1 transcriptional repressor (gene ID 2672). Gfi1 is also known as SCN2; GFI-1; GFI1A; ZNF163. This gene encodes a nuclear zinc finger protein that functions as a transcriptional repressor. This protein plays a role in diverse developmental contexts, including hematopoiesis and oncogenesis. Exemplary human nucleic acid and amino acid sequences are represented by the NCBI reference sequences NM_001127215.1 and NP_001120687.1 respectively.
- Accordingly, an “inhibitor of Gfi1” refers to a compound, substance or composition that can inhibit the function of Gfi1. For example, the inhibitor can inhibit the expression or activity of Gfi1. Examples of Gfi1 inhibitors include but are not limited to polypeptides such as dominant-negative protein mutants, peptidomimetics, antibodies, ribozymes, antisense oligonucleotides, or other small organic molecules which specifically inhibit the activity or expression of Gfi1.
- The term “small organic molecule” refers to a molecule of a size comparable to those organic molecules generally used in pharmaceuticals. The term excludes biological macromolecules (e.g., proteins, nucleic acids, etc.). Preferred small organic molecules range in size up to about 5000 Da, more preferably up to 2000 Da, and most preferably up to about 1000 Da. In a particular embodiment, the small molecule is T-3775440 as described in Ishikawa et al 2016. This molecule has the following structure in the art:
- In some embodiments, the inhibitor is a short hairpin RNA (shRNA), a small interfering RNA (siRNA) or an antisense oligonucleotide which inhibits the expression of Gfi1. A short hairpin RNA (shRNA) is a sequence of RNA that makes a tight hairpin turn that can be used to silence gene expression via RNA interference. shRNA is generally expressed using a vector introduced into cells, wherein the vector utilizes the U6 promoter to ensure that the shRNA is always expressed. This vector is usually passed on to daughter cells, allowing the gene silencing to be inherited. The shRNA hairpin structure is cleaved by the cellular machinery into siRNA, which is then bound to the RNA-induced silencing complex (RISC). This complex binds to and cleaves mRNAs that match the siRNA to which it is bound. Small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, are a class of 20-25 nucleotide-long double-stranded RNA molecules that play a variety of roles in biology. Most notably, siRNA is involved in the RNA interference (RNAi) pathway whereby the siRNA interferes with the expression of a specific gene. Anti-sense oligonucleotides include anti-sense RNA molecules and anti-sense DNA molecules, would act to directly block the translation of the targeted mRNA by binding thereto and thus preventing protein translation or increasing mRNA degradation, thus decreasing the level of the targeted protein, and thus activity, in a cell. For example, antisense oligonucleotides of at least about 15 bases and complementary to unique regions of the mRNA transcript sequence can be synthesized, e.g., by conventional phosphodiester techniques. Methods for using antisense techniques for specifically inhibiting gene expression of genes whose sequence is known are well known in the art (e.g. see U.S. Pat. Nos. 6,566,135; 6,566,131; 6,365,354; 6,410,323; 6,107,091; 6,046,321; and 5,981,732). Antisense oligonucleotides, siRNAs, shRNAs of the invention may be delivered in vivo alone or in association with a vector. In its broadest sense, a “vector” is any vehicle capable of facilitating the transfer of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid to the cells and typically mast cells. Typically, the vector transports the nucleic acid to cells with reduced degradation relative to the extent of degradation that would result in the absence of the vector. In general, the vectors useful in the invention include, but are not limited to, plasmids, phagemids, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the antisense oligonucleotide, siRNA, shRNA or ribozyme nucleic acid sequences. Viral vectors are a preferred type of vector and include, but are not limited to nucleic acid sequences from the following viruses: retrovirus, such as moloney murine leukemia virus, harvey murine sarcoma virus, murine mammary tumor virus, and rous sarcoma virus; adenovirus, adeno-associated virus; SV40-type viruses; polyoma viruses; Epstein-Barr viruses; papilloma viruses; herpes virus; vaccinia virus; polio virus; and RNA virus such as a retrovirus. One can readily employ other vectors not named but known to the art.
- In some embodiments, the inhibitor of Gif1 expression is an endonuclease. In the last few years, staggering advances in sequencing technologies have provided an unprecedentedly detailed overview of the multiple genetic aberrations in cancer. By considerably expanding the list of new potential oncogenes and tumor suppressor genes, these new data strongly emphasize the need of fast and reliable strategies to characterize the normal and pathological function of these genes and assess their role, in particular as driving factors during oncogenesis. As an alternative to more conventional approaches, such as cDNA overexpression or downregulation by RNA interference, the new technologies provide the means to recreate the actual mutations observed in cancer through direct manipulation of the genome. Indeed, natural and engineered nuclease enzymes have attracted considerable attention in the recent years. The mechanism behind endonuclease-based genome inactivating generally requires a first step of DNA single or double strand break, which can then trigger two distinct cellular mechanisms for DNA repair, which can be exploited for DNA inactivating: the errorprone nonhomologous end-joining (NHEJ) and the high-fidelity homology-directed repair (HDR). Endonucleases for gene inactivation have come in various forms, which includes CRISPR)/CRISPR associated (Cas) systems, mega nucleases (MN), zinc finger nucleases (ZFN), and transcription activator-like effector nucleases (TALEN). Endonucleases for use in the present invention are disclosed in WO 2010/079430, WO2011072246, WO2013045480, Mussolino C, et al (Curr Opin Biotechnol. 2012 October; 23(5):644-50) and Papaioannou I. et al (Expert Opinion on Biological Therapy, March 2012, Vol. 12, No. 3: 329-342).
- In a particular embodiment, the endonuclease is CRISPR-cas. As used herein, the term “CRISPR-cas” has its general meaning in the art and refers to clustered regularly interspaced short palindromic repeats associated which are the segments of prokaryotic DNA containing short repetitions of base sequences.
- In some embodiment, the endonuclease is CRISPR-cas9 which is from Streptococcus pyogenes. The CRISPR/Cas9 system has been described in U.S. Pat. No. 8,697,359 B1 and US 2014/0068797. Originally an adaptive immune system in prokaryotes (Barrangou and Marraffini, 2014), CRISPR has been recently engineered into a new powerful tool for genome editing. It has already been successfully used to target important genes in many cell lines and organisms, including human (Mali et al., 2013, Science, Vol. 339: 823-826), bacteria (Fabre et al., 2014, PLoS Negl. Trop. Dis., Vol. 8:e2671.), zebrafish (Hwang et al., 2013, PLoS One, Vol. 8:e68708.), C. elegans (Hai et al., 2014 Cell Res. doi: 10.1038/cr.2014.11.), bacteria (Fabre et al., 2014, PLoS Negl. Trop. Dis., Vol. 8:e2671.), plants (Mali et al., 2013, Science, Vol. 339: 823-826), Xenopus tropicalis (Guo et al., 2014, Development, Vol. 141: 707-714.), yeast (DiCarlo et al., 2013, Nucleic Acids Res., Vol. 41: 4336-4343.), Drosophila (Gratz et al., 2014 Genetics, doi:10.1534/genetics.113.160713), monkeys (Niu et al., 2014, Cell, Vol. 156: 836-843.), rabbits (Yang et al., 2014, J. Mol. Cell Biol., Vol. 6: 97-99.), pigs (Hai et al., 2014, Cell Res. doi: 10.1038/cr.2014.11.), rats (Ma et al., 2014, Cell Res., Vol. 24: 122-125.) and mice (Mashiko et al., 2014, Dev. Growth Differ. Vol. 56: 122-129.). Several groups have now taken advantage of this method to introduce single point mutations (deletions or insertions) in a particular target gene, via a single gRNA. Using a pair of gRNA-directed Cas9 nucleases instead, it is also possible to induce large deletions or genomic rearrangements, such as inversions or translocations. A recent exciting development is the use of the dCas9 version of the CRISPR/Cas9 system to target protein domains for transcriptional regulation, epigenetic modification, and microscopic visualization of specific genome loci.
- In some embodiment, the endonuclease is CRISPR-Cpf1 which is the more recently characterized CRISPR from Provotella and Francisella 1 (Cpf1) in Zetsche et al. (“Cpf1 is a Single RNA-guided Endonuclease of a
Class 2 CRISPR-Cas System (2015); Cell; 163, 1-13). - In some embodiments, the inhibitor is an intrabody having specificity for Gfi1. As used herein, the term “intrabody” generally refer to an intracellular antibody or antibody fragment. Antibodies, in particular single chain variable antibody fragments (scFv), can be modified for intracellular localization. Such modification may entail for example, the fusion to a stable intracellular protein, such as, e.g., maltose binding protein, or the addition of intracellular trafficking/localization peptide sequences, such as, e.g., the endoplasmic reticulum retention. In some embodiments, the intrabody is a single domain antibody. In some embodiments, the antibody according to the invention is a single domain antibody. The term “single domain antibody” (sdAb) or “VHH” refers to the single heavy chain variable domain of antibodies of the type that can be found in Camelid mammals which are naturally devoid of light chains. Such VHH are also called “Nanobody®”. According to the invention, sdAb can particularly be llama sdAb.
- As used herein the terms “administering” or “administration” refer to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., an inhibitor of Gif1) into the subject, such as by mucosal, intradermal, intravenous, subcutaneous, intramuscular delivery and/or any other method of physical delivery described herein or known in the art. When a disease, or a symptom thereof, is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof. When a disease or symptoms thereof, are being prevented, administration of the substance typically occurs before the onset of the disease or symptoms thereof.
- By a “therapeutically effective amount” of the inhibitor as above described is meant a sufficient amount to provide a therapeutic effect. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific polypeptide employed; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day. Typically, the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient. An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day.
- According to the invention, the inhibitor is administered to the subject in the form of a pharmaceutical composition. Typically, the inhibitor may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form therapeutic compositions. “Pharmaceutically” or “pharmaceutically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. In the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, the active principle, alone or in combination with another active principle, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings. Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms. Typically, the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The inhibitor can be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin. Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the typical methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The preparation of more, or highly concentrated solutions for direct injection is also contemplated, where the use of DMSO as solvent is envisioned to result in extremely rapid penetration, delivering high concentrations of the active agents. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed. For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
- In some embodiments, the inhibitor of the present invention is administered to the subject in combination with another diabetes medications. Exemplary diabetes medications include sulfonylureas, meglitinides, biguanides, thiazolidinediones, alpha-glucosidase inhibitors, or DPP-4 inhibitors. Sulfonylureas stimulate the beta cells of the pancreas to release more insulin. Chlorpropamide (Diabinese) is the only first-generation sulfonylurea still in use today. The second generation sulfonylureas are used in smaller doses than the first-generation drugs. There are three second-generation drugs: glipizide (Glucotrol and Glucotrol XL), glyburide (Micronase, Glynase, and Diabeta), and glimepiride (Amaryl). Meglitinides are drugs that also stimulate the beta cells to release insulin. Repaglinide (Prandin) and nateglinide (Starlix) are meglitinides. Metformin (Glucophage) is a biguanide. Biguanides lower blood glucose levels primarily by decreasing the amount of glucose produced by the liver. Rosiglitazone (Avandia) and pioglitazone (ACTOS) are in a group of drugs called thiazolidinediones. These drugs help insulin work better in the muscle and fat and also reduce glucose production in the liver. DPP-4 inhibitors help improve AIC without causing hypoglycemia. They work by preventing the breakdown of a naturally occurring compound in the body, GLP-1. GLP-1 reduces blood glucose levels in the body, but is broken down very quickly so it does not work well when injected as a drug itself. By interfering in the process that breaks down GLP-1, DPP-4 inhibitors allow it to remain active in the body longer, lowering blood glucose levels only when they are elevated. Sitagliptin (JANUVIA) and saxagliptin (ONGLYZA) are the two DPP-4 inhibitors currently on the market.
- A further object of the present invention relates to a method of screening a drug suitable for the treatment of hyperglycemia comprising i) providing a test compound and ii) determining the ability of said test compound to inhibit the expression or activity of Gfi1.
- Any biological assay well known in the art could be suitable for determining the ability of the test compound to inhibit the activity or expression of Gfi1. In some embodiments, the assay first comprises determining the ability of the test compound to bind to Gfi1. In some embodiments, a population of pancreatic cells (acinar, ductal or endocrine) is then contacted and activated so as to determine the ability of the test compound to inhibit the activity or expression of Gfi1. In particular, the effect triggered by the test compound is determined relative to that of a population of immune cells incubated in parallel in the absence of the test compound or in the presence of a control agent either of which is analogous to a negative control condition. The term “control substance”, “control agent”, or “control compound” as used herein refers a molecule that is inert or has no activity relating to an ability to modulate a biological activity or expression. It is to be understood that test compounds capable of inhibiting the activity or expression of Gfi1, as determined using in vitro methods described herein, are likely to exhibit similar modulatory capacity in applications in vivo. In vivo assays are well known in the art and typically include those described in the EXAMPLE. Typically, the test compound is selected from the group consisting of peptides, petptidomimetics, small organic molecules, antibodies (e.g. intraantibodies), aptamers or nucleic acids. For example the test compound according to the invention may be selected from a library of compounds previously synthesised, or a library of compounds for which the structure is determined in a database, or from a library of compounds that have been synthesised de novo. In some embodiments, the test compound may be selected form small organic molecules.
- The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.
-
FIG. 1 : qRT-PCR quantification of Gfi1 expression in mouse pancreas from E15.5 (embryonic day 15.5) to 11.8 months of age. -
FIG. 2 : X-gal staining and immunohistochemistry of Gfi1Cre::Rosa-lox--stop-lox-beta-Gal pancreas. -
FIG. 3 : Immunohistochemistry of adult pancreas labelled with insulin, amylase and DAPI. -
FIG. 4 : qRT-PCR assessment of amylase gene expression from whole pancreas at different developmental and adult stages. -
FIG. 5 : Immunohistochemical assessment of BrdU-labelled cells. -
FIG. 6 : Quantitative analyses of BrdU-labelled cells in different pancreatic compartments -
FIG. 7A-D : Intraperitoneal glucose tolerance test on 3 months old mice (A) and 6.5 months old mice (B). Intraperitoneal insulin tolerance test on 3 months old mice (C) and 6.5 months old mice (D). -
FIG. 8A-D : Weight and basal glycemia monitoring of control and Gfi1 mutant mice subjected to high fat diet (A-B). Glucose tolerance test (C) and insulin tolerance test (D) performed on mice of the indicated genotypes following exposure to high fat diet for 5 months. -
FIG. 9 : Basal glycaemia monitoring of control and Gfi1 mutant mice treated with high dose streptozotocin. Arrow represent days of streptozotocin injection. -
FIG. 10 : Insulin immunostaining ofpancreatic sections 5 days after acute streptozotocin treatment. -
FIG. 11 : qRT-PCR quantification of NKX6.2 mRNA levels from whole pancreas at different developmental and adult stages. -
FIG. 12 : RNA scope targeting NKX6.2 mRNA - Through a thorough quantitative analysis using qRT-PCR, we demonstrated that Gfi1 is expressed in the pancreas, starting from the first stages of pancreatic embryonic development. Furthermore, we observed that Gfi1 mRNA levels remain steady throughout embryonic development, while they significantly increase during the first days of life (from Postnatal day 6). A further increase was also noted between 1 and 3 months of age. Afterwards Gfi1 expression appeared to be stable during adulthood until at least 12 months of life (
FIG. 1 ). To investigate Gfi1 expression pattern, a Gfi1-Cre mouse line (expressing the phage P1 Cre Recombinase under the control of the endogenous Gfi1 promoter) was crossed with a ROSA26-β-gal mouse line (harboring a transgene containing the Rosa26 promoter upstream of a loxP-Neomycin resistance-Stop-loxP-β-galactosidase cassette). In the resulting Gfi1Cre::RosaLac animals, cells are permanently labelled once Gfi1 expression is activated. Thus, the spatiotemporal expression of β-galactosidase driven by Gfi1 promoter was analyzed by X-gal staining. β-galactosidase activity was solely detected in the acinar compartment. Importantly, no activity was found in ductal nor islets of Langerhans cells, demonstrating that the latter do not derive from cells having expressed Gfi1 (FIG. 2 ). - Gfi1 null animals suffer from several health conditions: loss of hearing, a general growth deficiency and severe neutropenia that impairs their life expectancy to a maximum of 1 month. To characterize the effects of Gfi1 ablation from the early phases of development to adulthood and thereafter, we therefore developed a conditional knock out system, where Gfi1 expression is silenced exclusively in the pancreas from the first phases of development. We thereby generated the Pdx1Cre::Gfi1cko transgenic mouse line (16). In the resulting animals, the Pdx1 promoter drives the expression of the Cre recombinase in all pancreatic cells. Once synthetized, the Cre recombinase induces Gfi1 loss-of-function solely in the pancreas. Pdx1Cre::Gfi1cko mice were found to be viable, fertile and to display a life expectancy comparable to that of control animals.
- Pdx1Cre::Gfi1cko pancreata were thoroughly analyzed by immunohistochemistry. Our phenotypical characterization showed that the majority of acinar cells were either negative or weakly positive for amylase (
FIG. 3 ). To confirm these observations, the amount of amylase transcript was assessed by RT-qPCR at different developmental and adult (st)ages. As expected, amylase expression levels increased significantly from E15.5 throughout postnatal life of control animals up to 3 months of age. Importantly, amylase transcript contents also raised continuously throughout pre- and post-natal development, but these were consistently decreased compared to controls at all ages tested. Strikingly, a significant 78.3% decrease in amylase expression level was measured in 3 months old Gfi1-mutants (FIG. 4 ). - Additionally, Gfi1 mutant acinar cells are hyperproliferative. Indeed, following 5 days of BrdU supplementation, BrdU immunoreactive cells were rarely observed in 3 months old control pancreata while an impressive number of dividing cells was detected in pancreatic sections isolated from adult Gfi1-deficient mice (
FIG. 5 ). Quantitative analyses demonstrated that the increase in the cellular proliferation rate was restricted to the acinar compartment (FIG. 6 ). - We extensively evaluated pancreas functionality, by performing several tests. We performed an intraperitoneal glucose tolerance test on 3-month old animals and noticed that the basal glycemia of Pdx1Cre::Gfi1cko was significantly lower, mutant mice displaying a better glucose tolerance compared to controls (
FIG. 7A ). However, no difference in insulin tolerance was found when we performed an intraperitoneal insulin tolerance test (FIG. 7C ). Importantly, the differences in glucose handling were more dramatic when we compared 6-month old Pdx1Cre::Gfi1cko mice with age matched controls (FIG. 7B ). Furthermore, at 6 months of age, Pdx1Cre::Gfi1cko mice were significantly more responsive to exogenous insulin administration compared to age matched controls (FIG. 7D ). Importantly, we challenged Pdx1Cre::Gfi1cko mice with high fat diet (33.3% of anhydrous butter and 16.6% of sucrose—a known model oftype 2 diabetes) for 5 months and monitored their weight and glycemia weekly. All the animals displayed a rapid increase in body mass as expected (FIG. 8A ). Surprisingly, while control mice rapidly developed a massive hyperglycemia, mutant mice remained normoglycemic (with a glycemia lower than 120 mg/dl) throughout the entire experiment (FIG. 8B ). At the end of the treatment, an intraperitoneal glucose tolerance test showed that Pdx1Cre::Gfi1cko mice were able to normally clear the blood of glucose in respect to the age-matched controls that showed an impaired capability (FIG. 8C ). Lastly, an intraperitoneal insulin tolerance test demonstrated that Gfi1 mutant animals did not develop any insulin resistance upon high fat diet as seen for their control counterparts (FIG. 8D ). - To determine whether Gfi1 loss could also be beneficial in Type I diabetes, we treated mutant and control mice with high dose streptozotocin to ablate their β-cells. Notably, 6 out of 9 controls responded to streptozotocin while all transgenic mice remained normoglycemic (
FIG. 9 ). After 4 days, we re-injected Pdx1Cre::Gfi1cko and non-responsive control mice with high dose streptozotocin: while all control animals rapidly developed acute hyperglycemia, Pdx1Cre::Gfi1cko mice remained either normoglycemic or developed a temporary mild hyperglycemia from which they quickly recovered from (FIG. 9 ). In a subsequent experiment, mice were sacrificed 5 days after streptozotocin injection and pancreas specimens were analyzed by immunohistochemistry. Surprisingly, the β-cell mass and islets of Langerhans integrity were found unaffected by streptozotocin cytotoxic activity in Gfi1 conditional knockout pancreata (FIG. 10 ). - To dissect the molecular pathway underlying Gfi1 activities in the pancreas, we analyzed the pancreatic transcriptome extracted from Gfi1-deficient mice by RT-qPCR and assessed the expression level of all genes known for their involvement in pancreas development and function. Using this approach, we found that Nkx6.2 was abnormally expressed in Pdx1Cre::Gfi1cko mice. Nkx6.2 transcripts were solely detected at E15.5 in control pancreas, Nkx6.2 being subsequently switched off (
FIG. 11 ). Importantly, Pdx1Cre::Gfi1cko pancreatic cells failed to inactivate Nkx6.2 expression during the latest stages of embryonic development (FIG. 11 ). Interestingly, by means of RNA scope, we detected Nkx6.2 mRNA molecules exclusively in the acinar cells (FIG. 12 ), suggesting a role of Nkx6.2 reactivation in the phenotype exhibited by Pdx1Cre::Gfi1cko mice. Together, our results indicate that the loss of Gfi1 in the pancreas results in a protection from all models of diabetes. Such protection originates in the acinar pancreas with appears altered with the loss of amylase, an hyperproliferation and a misexpression of Nkx6.2. All these evidence point to an altered absorption of glucose and role of Gfi1 in controlling beta-cell function. - Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.
- 1. Slack J M. Developmental biology of the pancreas. Development. 1995 June; 121(6):1569-80. PubMed PMID: 7600975.
- 2. Konstantinova I, Nikolova G, Ohara-Imaizumi M, Meda P, Kucera T, Zarbalis K, et al. EphA-Ephrin-A-mediated beta cell communication regulates insulin secretion from pancreatic islets. Cell. 2007 04/20; 129(2):359-70.
- 3. Prado C L, Pugh-Bernard A E, Elghazi L, Sosa-Pineda B, Sussel L. Ghrelin cells replace insulin-producing beta cells in two mouse models of pancreas development. Proceedings of the National Academy of Sciences of the United States of America. 2004 03/02; 101(9):2924-9.
- 4. Adrian T E, Bloom S R, Hermansen K, Iversen J. Pancreatic polypeptide, glucagon and insulin secretion from the isolated perfused canine pancreas. Diabetologia. 1978; 14(6):413-7.
- 5. Csaba Z, Dournaud P. Cellular biology of somatostatin receptors. Neuropeptides. 2001; 35(1):1-23.
- 6. Roncoroni L, Violi V, Montanari M, Muri M. Effect of somatostatin on exocrine pancreas evaluated on a total external pancreatic fistula of neoplastic origin. Am J Gastroenterol. 1983; 78(7):425-8.
- 7. Heller R S, Jenny M, Collombat P, Mansouri A, Tomasetto C, Madsen O D, et al. Genetic determinants of pancreatic epsilon-cell development. Developmental biology. 2005 10/01; 286(1):217-24.
- 8. Vignjevic S, Todorovic V, Damjanovic S, Budec M, Mitrovic O, Djikic D, et al. Similar developmental patterns of ghrelin- and glucagon-expressing cells in the human pancreas. Cells, tissues, organs. 2012; 196(4):362-73. PubMed PMID: 22538872.
- 9. Wierup N, Svensson H, Mulder H, Sundler F. The ghrelin cell: a novel developmentally regulated islet cell in the human pancreas. Regulatory peptides. 2002 Jul. 15; 107(1-3):63-9. PubMed PMID: 12137967.
- 10. Shih H P, Wang A, Sander M. Pancreas organogenesis: from lineage determination to morphogenesis. Annu Rev Cell Dev Biol. 2013; 29:81-105. PubMed PMID: 23909279.
- 11. Arda H E, Benitez C M, Kim S K. Gene regulatory networks governing pancreas development. Developmental cell. 2013 Apr. 15; 25(1):5-13. PubMed PMID: 23597482. Pubmed Central PMCID: 3645877.
- 12. Hock H, Orkin S H. Zinc-finger transcription factor Gfi-1: versatile regulator of lymphocytes, neutrophils and hematopoietic stem cells. Current opinion in hematology. 2006 January; 13(1):1-6. PubMed PMID: 16319680.
- 13. Linnoila R I, Jensen-Taubman S, Kazanjian A, Grimes H L. Loss of GFI1 impairs pulmonary neuroendorine cell proliferation, but the neuroendocrine phenotype has limited impact on post-naphthalene airway repair. Laboratory investigation; a journal of technical methods and pathology. 2007 April; 87(4):336-44. PubMed PMID: 17377622. Pubmed Central PMCID: 2839158.
- 14. Shroyer N F, Wallis D, Venken K J, Bellen H J, Zoghbi H Y. Gfi1 functions downstream of Math1 to control intestinal secretory cell subtype allocation and differentiation. Genes & development. 2005 Oct. 15; 19(20):2412-7. PubMed PMID: 16230531. Pubmed Central PMCID: 1257395.
- 15. Wallis D, Hamblen M, Zhou Y, Venken K J, Schumacher A, Grimes H L, et al. The zinc finger transcription factor Gfi1, implicated in lymphomagenesis, is required for inner ear hair cell differentiation and survival. Development. 2003 January; 130(1):221-32. PubMed PMID: 12441305.
- 16. Gannon M, Herrera P L, Wright C V. Mosaic Cre-mediated recombination in pancreas using the pdx-1 enhancer/promoter. Genesis. 2000 February; 26(2):143-4. PubMed PMID: 10686611.
- 17. Zhu J, Jankovic D, Grinberg A, Guo L, Paul W E. Gfi-1 plays an important role in IL-2-mediated Th2 cell expansion. Proceedings of the National Academy of Sciences of the United States of America. 2006 Nov. 28; 103(48):18214-9. PubMed PMID: 17116877. Pubmed Central PMCID: 1654136.
Claims (9)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/200,146 US20210290595A1 (en) | 2017-03-24 | 2021-03-12 | Gfi1 inhibitors for the treatment of hyperglycemia |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP17305334 | 2017-03-24 | ||
EP17305334.9 | 2017-03-24 | ||
PCT/EP2018/057680 WO2018172570A1 (en) | 2017-03-24 | 2018-03-26 | Gfi1 inhibitors for the treatment of hyperglycemia |
US201916496761A | 2019-09-23 | 2019-09-23 | |
US17/200,146 US20210290595A1 (en) | 2017-03-24 | 2021-03-12 | Gfi1 inhibitors for the treatment of hyperglycemia |
Related Parent Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/496,761 Continuation US10980779B2 (en) | 2017-03-24 | 2018-03-26 | GFI1 inhibitors for the treatment of hyperglycemia |
PCT/EP2018/057680 Continuation WO2018172570A1 (en) | 2017-03-24 | 2018-03-26 | Gfi1 inhibitors for the treatment of hyperglycemia |
Publications (1)
Publication Number | Publication Date |
---|---|
US20210290595A1 true US20210290595A1 (en) | 2021-09-23 |
Family
ID=58488940
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/496,761 Active US10980779B2 (en) | 2017-03-24 | 2018-03-26 | GFI1 inhibitors for the treatment of hyperglycemia |
US17/200,146 Abandoned US20210290595A1 (en) | 2017-03-24 | 2021-03-12 | Gfi1 inhibitors for the treatment of hyperglycemia |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/496,761 Active US10980779B2 (en) | 2017-03-24 | 2018-03-26 | GFI1 inhibitors for the treatment of hyperglycemia |
Country Status (3)
Country | Link |
---|---|
US (2) | US10980779B2 (en) |
EP (1) | EP3600269A1 (en) |
WO (1) | WO2018172570A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117866900A (en) * | 2022-10-10 | 2024-04-12 | 南京大学 | Humanized cell, animal model, construction method and application thereof |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6566131B1 (en) | 2000-10-04 | 2003-05-20 | Isis Pharmaceuticals, Inc. | Antisense modulation of Smad6 expression |
US6410323B1 (en) | 1999-08-31 | 2002-06-25 | Isis Pharmaceuticals, Inc. | Antisense modulation of human Rho family gene expression |
US6107091A (en) | 1998-12-03 | 2000-08-22 | Isis Pharmaceuticals Inc. | Antisense inhibition of G-alpha-16 expression |
US5981732A (en) | 1998-12-04 | 1999-11-09 | Isis Pharmaceuticals Inc. | Antisense modulation of G-alpha-13 expression |
US6046321A (en) | 1999-04-09 | 2000-04-04 | Isis Pharmaceuticals Inc. | Antisense modulation of G-alpha-i1 expression |
US6365354B1 (en) | 2000-07-31 | 2002-04-02 | Isis Pharmaceuticals, Inc. | Antisense modulation of lysophospholipase I expression |
US6566135B1 (en) | 2000-10-04 | 2003-05-20 | Isis Pharmaceuticals, Inc. | Antisense modulation of caspase 6 expression |
EP2206723A1 (en) | 2009-01-12 | 2010-07-14 | Bonas, Ulla | Modular DNA-binding domains |
EP2510096B2 (en) | 2009-12-10 | 2018-02-07 | Regents of the University of Minnesota | Tal effector-mediated dna modification |
EP2750671A2 (en) * | 2011-05-19 | 2014-07-09 | Oryzon Genomics, S.A. | Lysine demethylase inhibitors for thrombosis and cardiovascular diseases |
KR20140052018A (en) | 2011-08-09 | 2014-05-02 | 다케다 야쿠힌 고교 가부시키가이샤 | Cyclopropaneamine compound |
EP2573173B1 (en) | 2011-09-26 | 2015-11-11 | Justus-Liebig-Universität Gießen | Chimeric nucleases for gene targeting |
PE20150336A1 (en) | 2012-05-25 | 2015-03-25 | Univ California | METHODS AND COMPOSITIONS FOR RNA-DIRECTED MODIFICATION OF TARGET DNA AND FOR RNA-DIRECTED MODULATION OF TRANSCRIPTION |
US8697359B1 (en) | 2012-12-12 | 2014-04-15 | The Broad Institute, Inc. | CRISPR-Cas systems and methods for altering expression of gene products |
US9186391B2 (en) | 2013-08-29 | 2015-11-17 | Musc Foundation For Research Development | Cyclic peptide inhibitors of lysine-specific demethylase 1 |
-
2018
- 2018-03-26 US US16/496,761 patent/US10980779B2/en active Active
- 2018-03-26 WO PCT/EP2018/057680 patent/WO2018172570A1/en active Application Filing
- 2018-03-26 EP EP18715574.2A patent/EP3600269A1/en not_active Withdrawn
-
2021
- 2021-03-12 US US17/200,146 patent/US20210290595A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
EP3600269A1 (en) | 2020-02-05 |
US10980779B2 (en) | 2021-04-20 |
US20200030292A1 (en) | 2020-01-30 |
WO2018172570A1 (en) | 2018-09-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10443056B2 (en) | Methods of treating diabetes and/or promoting survival of pancreatic islets after transplantation | |
US10544415B2 (en) | Methods for producing enteroendocrine cells that make and secrete insulin | |
Gomez et al. | Experimental reconstitution of chronic ER stress in the liver reveals feedback suppression of BiP mRNA expression | |
US20140294924A1 (en) | Regeneration of Pancreatic Islets and Reversal of Diabetes by Islet Transcription Factor Genes Delivered In Vivo | |
EP2963108A1 (en) | Methods for inducing insulin production and uses thereof | |
JP2024020338A (en) | Method and composition for treating vitiligo | |
US20210290595A1 (en) | Gfi1 inhibitors for the treatment of hyperglycemia | |
EP2908853B1 (en) | Thy1 (cd90) as a therapy to control adipose tissue accumulation | |
CN110167564B (en) | Modulation of tjp1 expression to modulate cardiac cell regeneration | |
Ibrahim et al. | VIP as a potential therapeutic agent in gram negative sepsis | |
US20090305987A1 (en) | Method For Inducing Beta Cell Neogenesis From Epithelial Cells | |
Stamateris et al. | Noncanonical CDK4 signaling rescues diabetes in a mouse model by promoting β cell differentiation | |
CN116392500A (en) | micrornas and uses thereof in diagnosis and therapy | |
JP2022530031A (en) | Methods and compositions for miR-10 mimetics and their targets | |
JP2020515588A (en) | Methods for treating mitochondrial genetic disorders | |
Wideman | Manipulating proglucagon processing in the pancreatic alpha-cell for the treatment of diabetes | |
TW201139675A (en) | NeuroD1 gene expression in non-endocrine pancreatic epithelial cells (NEPECs) |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE (CNRS), FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:COLLOMBAT, PATRICK;NAPOLITANO, TIZIANA;AVOLIO, FABIO;SIGNING DATES FROM 20131007 TO 20131008;REEL/FRAME:055578/0041 Owner name: UNIVERSITE DE NICE SOPHIA ANTIPOLIS, FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:COLLOMBAT, PATRICK;NAPOLITANO, TIZIANA;AVOLIO, FABIO;SIGNING DATES FROM 20131007 TO 20131008;REEL/FRAME:055578/0041 Owner name: INSERM (INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE), FRANCE Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:COLLOMBAT, PATRICK;NAPOLITANO, TIZIANA;AVOLIO, FABIO;SIGNING DATES FROM 20131007 TO 20131008;REEL/FRAME:055578/0041 |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |