CN112851758B - 一种罗非鱼鳞促骨形成肽及其应用 - Google Patents
一种罗非鱼鳞促骨形成肽及其应用 Download PDFInfo
- Publication number
- CN112851758B CN112851758B CN202110113728.5A CN202110113728A CN112851758B CN 112851758 B CN112851758 B CN 112851758B CN 202110113728 A CN202110113728 A CN 202110113728A CN 112851758 B CN112851758 B CN 112851758B
- Authority
- CN
- China
- Prior art keywords
- tilapia mossambica
- bone formation
- peptide
- tilapia
- promoting peptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 56
- 230000001737 promoting effect Effects 0.000 title claims abstract description 46
- 241000276701 Oreochromis mossambicus Species 0.000 title claims abstract description 43
- 230000011164 ossification Effects 0.000 title claims abstract description 36
- 210000000963 osteoblast Anatomy 0.000 claims abstract description 28
- 230000033558 biomineral tissue development Effects 0.000 claims abstract description 15
- 230000004069 differentiation Effects 0.000 claims abstract description 15
- 230000035755 proliferation Effects 0.000 claims abstract description 14
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 208000001132 Osteoporosis Diseases 0.000 claims description 15
- 239000003814 drug Substances 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 16
- 230000009286 beneficial effect Effects 0.000 abstract description 7
- 231100000252 nontoxic Toxicity 0.000 abstract description 5
- 230000003000 nontoxic effect Effects 0.000 abstract description 5
- 241000276707 Tilapia Species 0.000 description 23
- 108060006698 EGF receptor Proteins 0.000 description 16
- 102000001301 EGF receptor Human genes 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 239000000243 solution Substances 0.000 description 12
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 11
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 11
- 210000000988 bone and bone Anatomy 0.000 description 10
- 238000003032 molecular docking Methods 0.000 description 9
- 102000008186 Collagen Human genes 0.000 description 8
- 108010035532 Collagen Proteins 0.000 description 8
- 229920001436 collagen Polymers 0.000 description 8
- 230000007935 neutral effect Effects 0.000 description 8
- 102000004196 processed proteins & peptides Human genes 0.000 description 7
- 238000005406 washing Methods 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000003993 interaction Effects 0.000 description 6
- 102000005701 Calcium-Binding Proteins Human genes 0.000 description 5
- 108010045403 Calcium-Binding Proteins Proteins 0.000 description 5
- 239000011575 calcium Substances 0.000 description 5
- 229910052791 calcium Inorganic materials 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000000589 high-performance liquid chromatography-mass spectrometry Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 239000006227 byproduct Substances 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000002209 hydrophobic effect Effects 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 2
- 101100335897 Caenorhabditis elegans gly-9 gene Proteins 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 2
- 238000010306 acid treatment Methods 0.000 description 2
- RGCKGOZRHPZPFP-UHFFFAOYSA-N alizarin Chemical compound C1=CC=C2C(=O)C3=C(O)C(O)=CC=C3C(=O)C2=C1 RGCKGOZRHPZPFP-UHFFFAOYSA-N 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000005238 degreasing Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 229940011871 estrogen Drugs 0.000 description 2
- 239000000262 estrogen Substances 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229960002901 sodium glycerophosphate Drugs 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000012192 staining solution Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- FQNILRVJOJBFFC-FXQIFTODSA-N Ala-Pro-Asp Chemical compound C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N FQNILRVJOJBFFC-FXQIFTODSA-N 0.000 description 1
- 241000269331 Ambystoma Species 0.000 description 1
- 241000269337 Ambystoma laterale Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 102000055006 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 description 1
- 102000043136 MAP kinase family Human genes 0.000 description 1
- 108091054455 MAP kinase family Proteins 0.000 description 1
- 208000030136 Marchiafava-Bignami Disease Diseases 0.000 description 1
- 208000029725 Metabolic bone disease Diseases 0.000 description 1
- 102100024193 Mitogen-activated protein kinase 1 Human genes 0.000 description 1
- PESQCPHRXOFIPX-UHFFFAOYSA-N N-L-methionyl-L-tyrosine Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 PESQCPHRXOFIPX-UHFFFAOYSA-N 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- LGMBKOAPPTYKLC-JYJNAYRXSA-N Pro-Phe-Arg Chemical compound C([C@@H](C(=O)N[C@@H](CCCNC(=N)N)C(O)=O)NC(=O)[C@H]1NCCC1)C1=CC=CC=C1 LGMBKOAPPTYKLC-JYJNAYRXSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 125000005340 bisphosphate group Chemical group 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 210000002805 bone matrix Anatomy 0.000 description 1
- 102000014823 calbindin Human genes 0.000 description 1
- 108060001061 calbindin Proteins 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- 229940069978 calcium supplement Drugs 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 229960001927 cetylpyridinium chloride Drugs 0.000 description 1
- YMKDRGPMQRFJGP-UHFFFAOYSA-M cetylpyridinium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+]1=CC=CC=C1 YMKDRGPMQRFJGP-UHFFFAOYSA-M 0.000 description 1
- 230000009920 chelation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000001360 collision-induced dissociation Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- 230000007783 downstream signaling Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 229940091249 fluoride supplement Drugs 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000324 molecular mechanic Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 102000014187 peptide receptors Human genes 0.000 description 1
- 108010011903 peptide receptors Proteins 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000004725 rapid separation liquid chromatography Methods 0.000 description 1
- 102000027426 receptor tyrosine kinases Human genes 0.000 description 1
- 108091008598 receptor tyrosine kinases Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 229910052712 strontium Inorganic materials 0.000 description 1
- CIOAGBVUUVVLOB-UHFFFAOYSA-N strontium atom Chemical compound [Sr] CIOAGBVUUVVLOB-UHFFFAOYSA-N 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Physical Education & Sports Medicine (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Animal Behavior & Ethology (AREA)
- Rheumatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Polymers & Plastics (AREA)
- Public Health (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明涉及一种罗非鱼鳞促骨形成肽及其应用。所述罗非鱼鳞促骨形成肽的氨基酸序列:APDPFRMY。本发明提供的罗非鱼鳞促骨形成肽具有较好的促进成骨细胞增殖活性,可促进成骨细胞ALP表达,有利于成骨细胞的分化和矿化,且安全无毒。
Description
技术领域
本发明属于生物材料技术领域,具体涉及一种罗非鱼鳞促骨形成肽及其应用。
背景技术
世界卫生组织(World Health Organization,WHO)将骨质疏松症定义为一种以机体骨质含量和骨质质量下降、骨组织纤维结构发生破坏、骨脆性增加且容易发生机体骨折为主要特征的全身性骨代谢性疾病。目前,全球骨质疏松患者人数已超2亿。我国是世界上骨质疏松人口数量最多的国家,预测到本世纪中叶,我国老龄人口将会超过4亿,骨量减少、骨质疏松患者将高达2亿。骨质疏松症已成为继糖尿病、心血管病之后的的第三大中老年慢性疾病。随着我国进入老龄化社会,人群骨质疏松症发病率也呈逐年增加的态势,骨质疏松症患者骨质发生疏松后导致的机体骨折严重增加了患者的病残率和病死率,己成为严重的社会公众健康问题。现阶段,临床上治疗骨质疏松症仍以化学药物为主,如双磷酸盐、雌激素、降钙素、氟化物、锶制剂药物。然而这些药物的临床治疗效果并不佳,长期的服用会带来副作用,如雌激素长期使用会增加女性罹患乳腺癌和子宫癌的风险性。经研究发现,一些天然活性肽起到可以预防和治疗骨质疏松的效果。成骨细胞是骨形成细胞,对骨的生长,维持骨量平衡以及骨代谢和损伤后修复,都起着至关重要的作用。它负责着骨基质的合成、分泌以及矿化。故对于取代传统治疗骨质疏松的方法,寻找可以促进成骨细胞增殖的天然活性肽对预防治疗骨质疏松症尤为重要。表皮生长因子受体(EGFR)是最重要的受体酪氨酸激酶之一,是一种53-AA多肽。表皮生长因子(EGF)通过与EGFR的胞外区结合,诱导受体二聚化,激活ERK通路,刺激成骨细胞增殖分化。一些下游信号通路被EGFR酪氨酸激酶触发,包括MAPK,它在增殖、分化和凋亡等复杂细胞反应中发挥关键作用。因此,EGFR的活化对体外基质再生具有重要的影响。对可以诱导EGFR激活的生物活性肽被认为是可以促骨形成的指标。专利CN108586604A就公开了可诱导EGFR激活的促成骨生物活性肽。
我国是全球最大罗非鱼养殖加工国,2016年养殖产量达158万吨,约占同年全球总产量的13%,此中大部分罗非鱼被用于鱼片加工。据统计,罗非鱼加工过程中产生的副产物可占所用鱼体总质量的54%,然而我国目前对加工副产物合理利用率较低,不但造成了生物资源浪费,也为环境增加负担。因此,开发利用罗非鱼副产物具有丰富的原料基础和广阔的应用前景。
发明内容
本发明的目的在于克服现有关于罗非鱼骨单体肽的生物利用度的研究缺乏的缺陷或不足,提供一种罗非鱼鳞促骨形成肽。本发明提供的罗非鱼鳞促骨形成肽具有较好的促进成骨细胞增殖活性,可促进成骨细胞ALP表达,有利于成骨细胞的分化和矿化,且安全无毒。
本发明的另一目的在于提供上述罗非鱼鳞促骨形成肽在制备促进成骨细胞增殖、分化或矿化的药物中的应用。
本发明的另一目的在于提供上述罗非鱼鳞促骨形成肽在制备促进成骨细胞增殖、分化或矿化的功能食品或保健品中的应用。
为实现上述发明目的,本发明采用如下技术方案:
一种罗非鱼鳞促骨形成肽,所述罗非鱼鳞促骨形成肽的氨基酸序列:APDPFRMY(Ala-Pro-Asp-Pro-Phe-Arg-Met-Tyr)。
本发明提供的罗非鱼鳞促骨形成肽具有较好的促进成骨细胞增殖活性,可促进成骨细胞ALP表达,有利于成骨细胞的分化,且安全无毒。
优选地,所述罗非鱼鳞促骨形成肽的分子量为995.4534Da。
优选地,所述罗非鱼鳞促骨形成肽的质荷比为498.7344。
优选地,所述罗非鱼鳞促骨形成肽与EGFR的氢键作用包括ARG285、GLN8、HIS409、LYS322、SER324或THR378中的一种或几种。
优选地,所述罗非鱼鳞促骨形成肽与EGFR的疏水相互作用包括ALA286、ASN86、ASP323、ASP344、GLN408、GLY317、GLY343、GLY39、GLY9、HIS346、ILE316、ILE318、LEU325、LEU38、LYS407、MET87、SER11、SER342或THR406中的一种或几种。
罗非鱼鳞促骨形成肽可通过如下过程制备得到:
S1、将罗非鱼鳞用蒸馏水重复洗三次,把杂质去除;
S2、碱洗脱脂:将罗非鱼鳞放入0.1mol/L NaOH溶液(w/v1:10)搅拌24h,清洗至中性;
S3、脱钙处理:将罗非鱼鳞放入10%EDTA-2Na(pH7.2,w/v1:10)中,并置于4℃冷库中,期间进行搅拌处理5天,随后将鱼鳞清洗至中性后待用;
S4、酸处理:将脱钙后的罗非鱼鳞用质量分数4%的盐酸处理时间18h后水洗至中性,即得到罗非鱼鳞胶原蛋白;将水洗至中性的罗非鱼鳞胶原蛋白中水分滴干,随后置于55℃烘箱中烘干,取出置于密封袋中备用;
S5、酶解:利用木瓜蛋白酶进行酶解,料液比为8%,酶解时间为2h,酶底比为0.3%,pH为7,温度为60℃。
上述罗非鱼鳞促骨形成肽在制备促进成骨细胞增殖、分化或矿化的药物中的应用也在本发明的保护范围内。
优选地,所述罗非鱼鳞促骨形成肽在制备治疗骨质疏松症药物中的应用。
优选地,所述药物包括药学上可接受的盐、载体和/或赋形剂。
上述罗非鱼鳞促骨形成肽在制备促进成骨细胞增殖、分化或矿化的功能食品或保健品中的应用也在本发明的保护范围内。
优选地,所述罗非鱼鳞促骨形成肽在制备奶粉、液态乳制品、钙片及补钙制剂或口服液中的应用。
与现有技术相比,本发明具有如下有益效果:
本发明提供的罗非鱼鳞促骨形成肽具有较好的促进成骨细胞增殖活性,可促进成骨细胞ALP表达,有利于成骨细胞的分化和矿化,且安全无毒。
附图说明
图1为EGFR几何与对接盒(PDB ID:1IVO);
图2为APDPFRMY和EGFR之间的交互作用三维图;
图3为APDPFRMY和EGFR相互作用2D图;其中,绿色虚线:氢键相互作用;数:键长;红堡:疏水相互作用;
图4为不同浓度的APDPFRMY的增殖效果图;
图5为TPERYY的ALP酶活效果图;
图6为TPERYY的矿化效果图;
图7为TPERYY的肽谱图
具体实施方式
下面结合实施例进一步阐述本发明。这些实施例仅用于说明本发明而不用于限制本发明的范围。下例实施例中未注明具体条件的实验方法,通常按照本领域常规条件或按照制造厂商建议的条件;所使用的原料、试剂等,如无特殊说明,均为可从常规市场等商业途径得到的原料和试剂。本领域的技术人员在本发明的基础上所做的任何非实质性的变化及替换均属于本发明所要求保护的范围。
实施例1罗非鱼鳞促骨形成肽的分离制备
本实施例提供一种罗非鱼鳞促骨形成肽,其氨基酸序列为:APDPFRMY(Ala-Pro-Asp-Pro-Phe-Arg-Met-Tyr)。
该罗非鱼鳞促骨形成肽可通过如下制备方法制备得到:
(一)罗非鱼鳞的预处理
(1)将罗非鱼鳞用蒸馏水重复洗三次,把杂质去除。
(2)碱洗脱脂:将罗非鱼鳞放入0.1mol/L NaOH溶液(w/v1:10)搅拌24h,清洗至中性。
(3)脱钙处理:将罗非鱼鳞放入10%EDTA-2Na(pH7.2,w/v1:10)中,并置于4℃冷库中,期间进行搅拌处理5天,随后将鱼鳞清洗至中性后待用。
(4)酸处理:将脱钙后的罗非鱼鳞用质量分数4%的盐酸处理时间18h后水洗至中性,即得到罗非鱼鳞胶原蛋白;将水洗至中性的罗非鱼鳞胶原蛋白中水分滴干,随后置于55℃烘箱中烘干,取出置于密封袋中备用。
(二)罗非鱼鳞钙结合肽的酶解实验
在料液比,8%,酶底比0.3%,pH 7下60℃水浴酶解2h,100℃灭酶10mim后,4000r/min离心20min,取上清,冷冻干燥。
(三)罗非鱼鳞钙结合肽的富集
将冻干的罗非鱼鳞钙结合肽,加入蒸馏水形成5mg/mL体系,按肽钙比1:1加入无水氯化钙后,调节pH 8,47℃水浴下螯合42min,螯合反应结束后,加入反应体系5倍体积的无水乙醇,4000r/min离心20min,弃上清,取沉淀,冷冻干燥。
(1)质谱序列:将富集的罗非鱼鳞钙结合肽利用HPLC-MS/MS对罗非鱼鳞钙结合肽分析。
HPLC-MS/MS法处理过程及条件如下:将C18除盐柱在平衡液(50%ACN)中吹打5次,用Washing buffer(0.1%FA,2%ACN)洗柱5次,吸取样品,缓慢吹打10次,用Elutionbuffer(0.1%FA,60%ACN)洗脱样品,洗脱溶液转至新管,离心浓缩干燥,待做质谱分析。除盐后多肽样品经离心干燥后,重新溶解于Nano-LC流动相A(0.1%甲酸/水)中装瓶上样,进行在线LCMS分析。溶解后的样品以2μL的体积上样到nanoViper C18预柱上(3μm,),然后20ul体积冲洗脱盐。液相为Easy nLC 1200纳升液相系统(ThermoFisher,USA),样品在预柱上脱盐保留后再经分析柱分离,分析柱规格是C18反相色谱柱(Acclaim PepMap RSLC,75μm×25cm C18-2μm),实验所用梯度为60min内流动相B(80%乙腈,0.1%甲酸)由5%升高至38%。质谱采用ThermoFisher Q Exactive系统(ThermoFisher,USA)结合纳升喷雾Nano Flex离子源(ThermoFisher,USA),喷雾电压为1.9kV,离子传输管加热温度为275℃。质谱扫描方式为信息依赖的采集工作模式下(DDA,Data Dependent Analysis),一级质谱扫描分辨率为70000,扫描范围350-2000m/z,最大注入时间100ms。每次DDA循环下最多采集20个电荷为2+到5+的二级图谱,二级质谱离子最大注入时间为50ms。碰撞室能量(高能碰撞诱导解离,HCD)设设定为28eV,适用于所有前体离子,动态排除设置为25秒。质谱采集到的原始raw图谱文件,采用PEAKS Studio 8.5(Bioinformatics Solutions Inc.Waterloo,Canada)软件进行数据加工处理和检索分析。
用HPLC-MS/MS法对罗非鱼鳞钙结合肽的氨基酸序列进行测定得到鉴定了133条肽序列,分子质量范围519-3736Da,肽段长度为5-43,其中包括序列为APDPFRMY的多肽(如图7)。
实施例2分子对接筛选罗非鱼鳞促骨形成肽
(1)分子对接
EGFR几何图形(如图1)从RCSB蛋白数据库下载,PDD ID:1IVO。水离子和其他不相干的原子被删除,准备对接。用PyMol1.7构建肽段APDPFRMY及其它肽段,然后用UFF(万向力场)分子力学力场进行能量最小化,使用Avogadro软件。与AutoDock Vina 1.2进行分子对接,对接口袋定义为一个50A*50A*50A的盒子(图1),包括报道的所有活性位点。以上未提到的所有参数均设置为默认值。
(2)分子对接筛选单体
肽段APDPFRMY与EGFR结合的能量表见表1,根据结合能可知,APDPFRMY与EGFR结合非常稳定。
表1通过HPLC-MS/MS鉴定的罗非鱼鳞钙结肽AutoDock Vina分子对接分析
(3)分子对接单体分析:
APDPFRMY与EGFR的氢键作用(如图2和图3)包括ARG285、GLN8、HIS409、LYS322、SER324和THR378。APDPFRMY与EGFR的疏水相互作用包括ALA286、ASN86、ASP323、ASP344、GLN408、GLY317、GLY343、GLY39、GLY9、HIS346、ILE316、ILE318、LEU325、LEU38、LYS407、MET87、SER11、SER342和THR406。
(4)单体合成
从Synpeptide Co.,Ltd(南京,中国)获得纯度高于95%的合成肽APDPFRMY。
实施例3罗非鱼鳞促骨形成肽的活性验证
(1)成骨细胞培养:
MC3T3-E1 subclone14细胞购自中国科学院上海生命科学研究院细胞资源中心。细胞在完全培养基中培养(α-MEM中添加10%胎牛血清,1%双抗)。细胞融汇80%-90%时,用胰蛋白酶-EDTA传代。
(2)细胞增殖率:
不同浓度的APDPFRMY对MC3T3-E1毒性试验。将MC3T3-E1细胞以5×103个细胞/孔的密度种植在96孔板(Costar,Corning,NY)中,并放在5%CO2培养箱中于37℃温育24小时。随后,用具有不同浓度(0、1、10、50、100、200μg/mL)的肽(APDPFRMY)的100μL培养基处理细胞,并孵育24小时。孵育后,通过100μL的0.5mg/mL MTT溶液处理细胞4小时。然后,用150μL二甲基亚砜代替MTT溶液。将96孔板置于振荡器上15min。最后,使用酶标仪在570nm的波长下测量光吸收值。与对照组相比,吸光度降低10%以上的样品浓度被认为是有细胞毒性的。
图4可知,在浓度范围为1,10,50,100,200μg/mL,APDPFRMY对细胞活力没有负面影响,根据MTT试验结果,选用0,1,10,50μg/mL进行分化矿化实验。
(3)碱性磷酸酶(ALP)活性:
将MC3T3-E1细胞以1×106个细胞/孔的密度种植在6孔板(Costar,Corning,NY)中,待细胞融合后,分别更换含不同浓度(0,1,10,50μg/mL)的APDPFRMY的分化完全培养液(含50μg/mL VC和10mmol/Lβ-甘油磷酸钠)培养7天。第七天后,根据ALP试剂盒(南京建成)进行ALP活性检测。
如图5,相对于空白对照,含有1,10,50μg/mL APDPFRMY的样品组碱性磷酸酶活都显著提高,说明APDPFRMY可以促进成骨细胞ALP表达,有利于成骨细胞的分化。
(4)茜素红染色:
将MC3T3-E1细胞以1×106个细胞/孔的密度种植在6孔板(Costar,Corning,NY)中,待细胞融合后,分别更换含不同浓度(0,1,10,50μg/mL)的APDPFRMY的分化完全培养液(含50μg/mL VC和10mmol/Lβ-甘油磷酸钠)培养21天,前14天隔天换液,后7天每天半量换液。21天后,弃去培养架,用PBS洗细胞2次,用4%多聚甲醛固定30min,弃甲醛,蒸馏水洗2次,加入1mL茜素红染液染色5min,弃去染液,蒸馏水洗3次,PBS在37℃10min除去特异性结合,拍照。利用10%的西吡氯铵溶液,溶解钙结节,在酶标仪562nm处进行钙含量测定。
图6显示了APDPFRMY对MC3T3-E1矿化有显著的影响。相对于空白组,含有1,10,50μg/mL APDPFRMY成骨细胞矿化结节都有不同程度的增加;在钙结节定量分析中,可以直观看出,相对于空白对照,含有10μg/mL APDPFRMY样品组矿化效果好,结果说明APDPFRMY可以促进成骨细胞矿化。
由上述可知,本发明提供的罗非鱼鳞促骨形成肽具有较好的促进成骨细胞增殖活性,可促进成骨细胞ALP表达,有利于成骨细胞的分化和矿化,且安全无毒。
最后应当指出的是,以上实施例仅是本发明的具有代表性的例子。显然,本发明的技术方案并不限于上述实施例,还可有许多变形。本领域的普通技术人员能从本发明内容直接导出或联想到所有变形,均应认为是本发明的权利要求的保护范围。
序列表
<110> 华南农业大学
<120> 一种罗非鱼鳞促骨形成肽及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 8
<212> PRT
<213> 罗非鱼鳞促骨形成肽(2 Ambystoma laterale x Ambystomajeffersonianum)
<400> 1
Ala Pro Asp Pro Phe Arg Met Tyr
1 5
Claims (4)
1.一种罗非鱼鳞促骨形成肽,其特征在于,所述罗非鱼鳞促骨形成肽的氨基酸序列为:APDPFRMY。
2.权利要求1所述罗非鱼鳞促骨形成肽在制备促进成骨细胞增殖、分化或矿化的药物中的应用。
3.根据权利要求2所述应用,其特征在于,所述罗非鱼鳞促骨形成肽在制备治疗骨质疏松症药物中的应用。
4.根据权利要求2所述应用,其特征在于,所述药物包括药学上可接受的盐、载体和/或赋形剂。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110113728.5A CN112851758B (zh) | 2021-01-27 | 2021-01-27 | 一种罗非鱼鳞促骨形成肽及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110113728.5A CN112851758B (zh) | 2021-01-27 | 2021-01-27 | 一种罗非鱼鳞促骨形成肽及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112851758A CN112851758A (zh) | 2021-05-28 |
CN112851758B true CN112851758B (zh) | 2022-03-22 |
Family
ID=75986135
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110113728.5A Active CN112851758B (zh) | 2021-01-27 | 2021-01-27 | 一种罗非鱼鳞促骨形成肽及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112851758B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101297673A (zh) * | 2008-05-15 | 2008-11-05 | 北京盛美诺生物技术有限公司 | 鱼胶原低聚肽的加工方法 |
JP2013107850A (ja) * | 2011-11-21 | 2013-06-06 | Meiji Co Ltd | コラーゲンペプチドの製造方法 |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR101131223B1 (ko) * | 2010-07-30 | 2012-03-28 | 부경대학교 산학협력단 | 나일 틸라피아로부터 분리된 항산화 펩티드 및 이를 포함하는 항산화 조성물 |
CN102172393A (zh) * | 2010-12-17 | 2011-09-07 | 中国海洋大学 | 一种鱼鳞蛋白肽钙螯合物补钙剂及其制备方法 |
GB201308828D0 (en) * | 2013-03-12 | 2013-07-03 | Verenium Corp | Phytase |
CN109258916A (zh) * | 2018-11-28 | 2019-01-25 | 潍坊医学院 | 一种鱼鳞胶原肽钙及其制备方法、用途 |
CN110627896B (zh) * | 2019-09-03 | 2022-02-18 | 华南农业大学 | 一种钙螯合肽及其制备方法和应用 |
CN111621537B (zh) * | 2020-05-09 | 2022-06-14 | 华中农业大学 | 一种淡水鱼鱼鳞胶原蛋白肽粉的制备方法 |
-
2021
- 2021-01-27 CN CN202110113728.5A patent/CN112851758B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101297673A (zh) * | 2008-05-15 | 2008-11-05 | 北京盛美诺生物技术有限公司 | 鱼胶原低聚肽的加工方法 |
JP2013107850A (ja) * | 2011-11-21 | 2013-06-06 | Meiji Co Ltd | コラーゲンペプチドの製造方法 |
Also Published As
Publication number | Publication date |
---|---|
CN112851758A (zh) | 2021-05-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Shi et al. | A bovine lactoferrin–derived peptide induced osteogenesis via regulation of osteoblast proliferation and differentiation | |
CN110655556B (zh) | 一种免疫调节肽的制备及方法 | |
CN108586604A (zh) | 促成骨生物活性肽及其筛选方法 | |
CN107602688A (zh) | 牛乳αs2‑酪蛋白来源生物活性肽的制备和应用 | |
CN102389440A (zh) | 环烯醚萜类化合物在制备抗骨质疏松药物中的应用 | |
Liu et al. | The effect of different molecular weight collagen peptides on MC3T3-E1 cells differentiation | |
CN108676073B (zh) | 一种抗肥胖十肽llvvypwtqr及其应用 | |
CN104987358B (zh) | 一种鹿茸蛋白提取物的制备方法 | |
Wang et al. | The separation of antler polypeptide and its effects on the proliferation and osteogenetic differentiation of bone marrow mesenchymal stem cells | |
CN112851758B (zh) | 一种罗非鱼鳞促骨形成肽及其应用 | |
CN104987359B (zh) | 一种鹿茸蛋白提取物及其应用、药物制剂 | |
CN111269292B (zh) | 具有促进组织修复的家蝇多肽及其制备方法与应用 | |
CN112679581B (zh) | 一种罗非鱼鳞促骨形成肽及其应用 | |
CN106749524B (zh) | 一种抗肥胖七肽npvwkrk | |
CN114874291B (zh) | 一种人工虎骨多肽及其制备方法和应用 | |
US11951155B2 (en) | Pea-derived peptide with muscle-building effect and preparation method thereof, and drug and use | |
US10342854B2 (en) | Leptin active peptide having cd loop and E helix region mutations, coding gene thereof, and application thereof | |
US20180050093A1 (en) | Leptin active peptide having d helix region mutations, coding gene thereof, and application thereof | |
CN115785219A (zh) | 具有缓解代谢综合征的谷糠活性肽及其制备方法与应用 | |
Zhong et al. | Effect of the phosphorylation structure in casein phosphopeptides on the proliferation, differentiation, and mineralization of osteoblasts and its mechanism | |
CN112956700A (zh) | 促骨形成肽在制备促进成骨细胞增殖、分化或矿化的功能性食品、保健品或药物中的应用 | |
CN100417662C (zh) | 无指盘臭蛙胰岛素释放促进肽和在制药中的应用 | |
CN110100945B (zh) | 一种汉麻降血脂肽组合物及其应用 | |
CN106749533B (zh) | 一种抗肥胖十七肽lnnpsvcdcdcmmkaar | |
CN111285922B (zh) | 一种促进组织修复的果蝇多肽及其制备方法与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |