CN112451652A - Recombinant human thyrotropin injection - Google Patents
Recombinant human thyrotropin injection Download PDFInfo
- Publication number
- CN112451652A CN112451652A CN202011417349.7A CN202011417349A CN112451652A CN 112451652 A CN112451652 A CN 112451652A CN 202011417349 A CN202011417349 A CN 202011417349A CN 112451652 A CN112451652 A CN 112451652A
- Authority
- CN
- China
- Prior art keywords
- present application
- pharmaceutical formulation
- thyrotropin
- concentration
- thyroid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108010061174 Thyrotropin Proteins 0.000 title claims abstract description 177
- 102000011923 Thyrotropin Human genes 0.000 title claims abstract description 177
- 230000001748 thyrotropin Effects 0.000 title claims abstract description 177
- 229960000874 thyrotropin Drugs 0.000 title claims abstract description 174
- 238000002347 injection Methods 0.000 title description 36
- 239000007924 injection Substances 0.000 title description 36
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 225
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims abstract description 88
- 229930182817 methionine Natural products 0.000 claims abstract description 84
- 238000000034 method Methods 0.000 claims abstract description 63
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 86
- 210000001685 thyroid gland Anatomy 0.000 claims description 68
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 46
- 229930195725 Mannitol Natural products 0.000 claims description 46
- 239000000594 mannitol Substances 0.000 claims description 46
- 235000010355 mannitol Nutrition 0.000 claims description 46
- 239000011780 sodium chloride Substances 0.000 claims description 43
- XUIIKFGFIJCVMT-GFCCVEGCSA-N D-thyroxine Chemical compound IC1=CC(C[C@@H](N)C(O)=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-GFCCVEGCSA-N 0.000 claims description 36
- 229940034208 thyroxine Drugs 0.000 claims description 36
- XUIIKFGFIJCVMT-UHFFFAOYSA-N thyroxine-binding globulin Natural products IC1=CC(CC([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-UHFFFAOYSA-N 0.000 claims description 36
- 208000024799 Thyroid disease Diseases 0.000 claims description 33
- 238000003745 diagnosis Methods 0.000 claims description 19
- 238000011282 treatment Methods 0.000 claims description 19
- 239000003814 drug Substances 0.000 claims description 14
- 238000002360 preparation method Methods 0.000 claims description 11
- 239000003381 stabilizer Substances 0.000 claims description 11
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 239000006172 buffering agent Substances 0.000 claims description 6
- 239000008177 pharmaceutical agent Substances 0.000 abstract 1
- 229960004452 methionine Drugs 0.000 description 81
- 210000004027 cell Anatomy 0.000 description 53
- 238000012360 testing method Methods 0.000 description 48
- 239000000872 buffer Substances 0.000 description 43
- 239000000047 product Substances 0.000 description 31
- 230000000694 effects Effects 0.000 description 27
- 239000000203 mixture Substances 0.000 description 27
- 208000015799 differentiated thyroid carcinoma Diseases 0.000 description 24
- 230000003647 oxidation Effects 0.000 description 23
- 238000007254 oxidation reaction Methods 0.000 description 23
- 235000018102 proteins Nutrition 0.000 description 23
- 102000004169 proteins and genes Human genes 0.000 description 23
- 108090000623 proteins and genes Proteins 0.000 description 23
- 238000003998 size exclusion chromatography high performance liquid chromatography Methods 0.000 description 22
- 238000003860 storage Methods 0.000 description 20
- 239000012669 liquid formulation Substances 0.000 description 19
- 239000000243 solution Substances 0.000 description 19
- 230000004071 biological effect Effects 0.000 description 18
- 239000000126 substance Substances 0.000 description 17
- 238000004458 analytical method Methods 0.000 description 16
- 230000006870 function Effects 0.000 description 16
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 15
- 238000001514 detection method Methods 0.000 description 15
- 239000004094 surface-active agent Substances 0.000 description 15
- 230000015572 biosynthetic process Effects 0.000 description 14
- 238000009472 formulation Methods 0.000 description 14
- 239000008363 phosphate buffer Substances 0.000 description 14
- 238000003786 synthesis reaction Methods 0.000 description 14
- 229920001213 Polysorbate 20 Polymers 0.000 description 13
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 13
- 239000011734 sodium Substances 0.000 description 13
- 230000003204 osmotic effect Effects 0.000 description 12
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 12
- 210000001519 tissue Anatomy 0.000 description 12
- 208000003532 hypothyroidism Diseases 0.000 description 10
- 230000002989 hypothyroidism Effects 0.000 description 10
- 239000007788 liquid Substances 0.000 description 10
- 229920001993 poloxamer 188 Polymers 0.000 description 10
- 229940044519 poloxamer 188 Drugs 0.000 description 10
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 10
- 229910000162 sodium phosphate Inorganic materials 0.000 description 9
- 229910019142 PO4 Inorganic materials 0.000 description 8
- 239000013583 drug formulation Substances 0.000 description 8
- 230000002265 prevention Effects 0.000 description 8
- 241000699802 Cricetulus griseus Species 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 7
- 208000024770 Thyroid neoplasm Diseases 0.000 description 7
- 239000000539 dimer Substances 0.000 description 7
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 7
- 239000002552 dosage form Substances 0.000 description 7
- 229940079593 drug Drugs 0.000 description 7
- 210000001672 ovary Anatomy 0.000 description 7
- 230000028327 secretion Effects 0.000 description 7
- 230000001225 therapeutic effect Effects 0.000 description 7
- 201000002510 thyroid cancer Diseases 0.000 description 7
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 6
- 102000009843 Thyroglobulin Human genes 0.000 description 6
- 108010034949 Thyroglobulin Proteins 0.000 description 6
- 230000015556 catabolic process Effects 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 229910000397 disodium phosphate Inorganic materials 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 229920001983 poloxamer Polymers 0.000 description 6
- 229920000136 polysorbate Polymers 0.000 description 6
- 229960002175 thyroglobulin Drugs 0.000 description 6
- 208000021510 thyroid gland disease Diseases 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 206010018498 Goitre Diseases 0.000 description 5
- 208000001204 Hashimoto Disease Diseases 0.000 description 5
- 208000030836 Hashimoto thyroiditis Diseases 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 5
- 201000003872 goiter Diseases 0.000 description 5
- 239000007972 injectable composition Substances 0.000 description 5
- 239000000825 pharmaceutical preparation Substances 0.000 description 5
- 229960000502 poloxamer Drugs 0.000 description 5
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- FFEARJCKVFRZRR-UHFFFAOYSA-N L-Methionine Natural products CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 4
- 229930195722 L-methionine Natural products 0.000 description 4
- 241000124008 Mammalia Species 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 230000036770 blood supply Effects 0.000 description 4
- 239000007857 degradation product Substances 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 229910052740 iodine Inorganic materials 0.000 description 4
- 239000011630 iodine Substances 0.000 description 4
- 230000007774 longterm Effects 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 4
- 229920001451 polypropylene glycol Polymers 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 238000012430 stability testing Methods 0.000 description 4
- 238000001356 surgical procedure Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 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 3
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 3
- 206010027476 Metastases Diseases 0.000 description 3
- 102000003992 Peroxidases Human genes 0.000 description 3
- 102000003673 Symporters Human genes 0.000 description 3
- 108090000088 Symporters Proteins 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 208000010928 autoimmune thyroid disease Diseases 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 108010068678 beta Subunit Thyrotropin Proteins 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 238000002270 exclusion chromatography Methods 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- -1 hydrogen ions Chemical class 0.000 description 3
- 230000026045 iodination Effects 0.000 description 3
- 238000006192 iodination reaction Methods 0.000 description 3
- 230000009401 metastasis Effects 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 108040007629 peroxidase activity proteins Proteins 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 208000020016 psychiatric disease Diseases 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 3
- 102000055006 Calcitonin Human genes 0.000 description 2
- 108060001064 Calcitonin Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 229920001214 Polysorbate 60 Polymers 0.000 description 2
- 208000028017 Psychotic disease Diseases 0.000 description 2
- 102000003911 Thyrotropin Receptors Human genes 0.000 description 2
- 108090000253 Thyrotropin Receptors Proteins 0.000 description 2
- WTZUXUQXRVKZCY-FHNDMYTFSA-M [Na+].[Cl-].CSCC[C@H](N)C(O)=O Chemical compound [Na+].[Cl-].CSCC[C@H](N)C(O)=O WTZUXUQXRVKZCY-FHNDMYTFSA-M 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 238000009098 adjuvant therapy Methods 0.000 description 2
- 108010000732 alpha Subunit Glycoprotein Hormones Proteins 0.000 description 2
- 102000002287 alpha Subunit Glycoprotein Hormones Human genes 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000001363 autoimmune Effects 0.000 description 2
- 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 2
- 229960004015 calcitonin Drugs 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000011221 initial treatment Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- XMBWDFGMSWQBCA-RNFDNDRNSA-M iodine-131(1-) Chemical compound [131I-] XMBWDFGMSWQBCA-RNFDNDRNSA-M 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 239000002736 nonionic surfactant Substances 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 230000001817 pituitary effect Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 229950008882 polysorbate Drugs 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000013558 reference substance Substances 0.000 description 2
- 230000000087 stabilizing effect Effects 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- SWMDCXYSYSMCJM-UHFFFAOYSA-N 2-aminopentanethioic s-acid Chemical compound CCCC(N)C(S)=O SWMDCXYSYSMCJM-UHFFFAOYSA-N 0.000 description 1
- XZIIFPSPUDAGJM-UHFFFAOYSA-N 6-chloro-2-n,2-n-diethylpyrimidine-2,4-diamine Chemical class CCN(CC)C1=NC(N)=CC(Cl)=N1 XZIIFPSPUDAGJM-UHFFFAOYSA-N 0.000 description 1
- 208000031873 Animal Disease Models Diseases 0.000 description 1
- 241000272517 Anseriformes Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000700199 Cavia porcellus Species 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010086677 Gonadotropins Proteins 0.000 description 1
- 102000006771 Gonadotropins Human genes 0.000 description 1
- 241000282575 Gorilla Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241000282620 Hylobates sp. Species 0.000 description 1
- 241000282553 Macaca Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000282405 Pongo abelii Species 0.000 description 1
- 102000029797 Prion Human genes 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 108010066702 Thyrotropin Alfa Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 238000011360 adjunctive therapy Methods 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 238000011558 animal model by disease Methods 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 239000003945 anionic surfactant Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 239000003093 cationic surfactant Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 229940114081 cinnamate Drugs 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000005750 disease progression Effects 0.000 description 1
- DGLRDKLJZLEJCY-UHFFFAOYSA-L disodium hydrogenphosphate dodecahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.O.O.[Na+].[Na+].OP([O-])([O-])=O DGLRDKLJZLEJCY-UHFFFAOYSA-L 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 238000009093 first-line therapy Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000002622 gonadotropin Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 125000001183 hydrocarbyl group Chemical group 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 125000003010 ionic group Chemical group 0.000 description 1
- 239000002563 ionic surfactant Substances 0.000 description 1
- 230000000366 juvenile effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000011866 long-term treatment Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000037323 metabolic rate Effects 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 230000007658 neurological function Effects 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 239000007793 ph indicator Substances 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229940068977 polysorbate 20 Drugs 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 235000013594 poultry meat Nutrition 0.000 description 1
- 229940071643 prefilled syringe Drugs 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000002731 protein assay Methods 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- BBMHARZCALWXSL-UHFFFAOYSA-M sodium dihydrogenphosphate monohydrate Chemical compound O.[Na+].OP(O)([O-])=O BBMHARZCALWXSL-UHFFFAOYSA-M 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000000534 thyroid cartilage Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 210000003437 trachea Anatomy 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M trans-cinnamate Chemical compound [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 229920000428 triblock copolymer Polymers 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000002676 xenobiotic agent Substances 0.000 description 1
- 230000002034 xenobiotic effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/14—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4
Abstract
The present application relates to a pharmaceutical formulation comprising thyrotropin and methionine, wherein the methionine is present at a concentration of about 0.005mg/mL or greater. Kits containing the pharmaceutical formulations, methods of making the pharmaceutical formulations, and uses of the pharmaceutical agents are also provided.
Description
Technical Field
The application relates to the field of biomedicine, in particular to a recombinant human thyrotropin injection.
Background
Currently, Differentiated Thyroid Cancer (DTC) has a high incidence rate, but the prognosis from surgical treatment is optimistic. After surgery for differentiated thyroid cancer, long-term treatment for inhibition of Thyrotropin (TSH) is effective in reducing the risk of tumor recurrence and controlling disease progression.
In recent years, natural human thyrotropin has been replaced by synthetic recombinant human thyrotropin. However, since TSH is a glycoprotein molecule, its biological activity is easily affected by environmental factors, and the synthesis and storage conditions are very harsh. The cost of treatment with TSH is therefore extremely high. Currently, commercial recombinant human thyrotropin products are provided in powder dosage form for administration as a solution by a physician or pharmacist prior to use. However, the oxidation speed of the recombinant human thyrotropin solution is very high, the recombinant human thyrotropin solution can only be prepared at present, the biological activity of the recombinant human thyrotropin solution is difficult to guarantee after the recombinant human thyrotropin solution is prepared, and the storage difficulty of the recombinant human thyrotropin increases the medication cost of patients.
Therefore, there is a need in the art for a recombinant human thyrotropin formulation with stable biological activity and a method for preparing the same.
Disclosure of Invention
The application provides a recombinant human thyrotropin preparation with stable biological activity and a preparation method thereof.
The recombinant human thyrotropin pharmaceutical formulations of the present application may have one or more of the following group of effects: (1) has storage stability; (2) the antioxidant property is provided; (3) has resistance to degradation; (4) the safety is high; (5) has higher content of thyrotropin stabilizing monomer; (6) with a lower content of degradation products, oxidation products and/or aggregates.
In one aspect, the present application provides a pharmaceutical formulation comprising thyrotropin and methionine, wherein the methionine is at a concentration of about 0.005mg/mL or greater.
In certain embodiments, the pharmaceutical formulation of the present application, wherein the methionine comprises L-methionine.
In certain embodiments, the pharmaceutical formulation of the present application, the methionine is at a concentration of about 0.005mg/mL to about 1.5 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the methionine is at a concentration of about 0.02mg/mL to about 1.5 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the methionine is at a concentration of about 0.005mg/mL, about 0.02mg/mL, about 0.1mg/mL, about 0.2mg/mL, or about 1.5 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the methionine is at a concentration of about 0.1 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the thyrotropin is selected from the group consisting of: thyrotropin isolated from mammals, and recombinant mammalian thyrotropin.
In certain embodiments, the pharmaceutical formulation of the present application, the thyrotropin comprises a cell-expressed recombinant human thyrotropin.
In certain embodiments, the pharmaceutical formulation of the present application, the thyrotropin comprises a mammalian cell-expressed recombinant human thyrotropin.
In certain embodiments, the pharmaceutical formulation of the present application, the thyrotropin comprises recombinant human thyrotropin expressed by Chinese Hamster Ovary (CHO) cells.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of thyrotropin is from about 0.45mg/mL to about 1.8 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of thyrotropin is from about 0.9mg/mL to about 1.8 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of thyrotropin is about 0.45mg/mL, about 0.9mg/mL, or about 1.8 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of thyrotropin is about 0.9 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application has a pH between about 6.5 and about 7.5.
In certain embodiments, the pharmaceutical formulation of the present application has a pH between about 6.5 and about 7.0.
In certain embodiments, the pharmaceutical formulation of the present application has a pH of about 6.5 or about 7.0.
In certain embodiments, the pharmaceutical formulation of the present application has a pH of about 7.0.
In certain embodiments, the pharmaceutical formulation of the present application further comprises a buffering agent.
In certain embodiments, the pharmaceutical formulation of the present application, the buffer comprises a phosphate buffer.
In certain embodiments, the pharmaceutical formulation of the present application, the buffer comprises a phosphate buffer having a pH between about 6.5 and about 7.5.
In certain embodiments, the pharmaceutical formulation of the present application, the buffer is at a concentration of about 20 mM.
In certain embodiments, the pharmaceutical formulation of the present application, the buffer has a pH of about 7.0.
In certain embodiments, the pharmaceutical formulation of the present application, the buffering agent comprises the group of: na (Na)2HPO4·12H2O, and NaH2PO4·H2O。
In certain embodiments, the pharmaceutical formulation of the present application, said Na2HPO4·12H2The concentration of O is from about 2.25mg/mL to about 6.82 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the NaH2PO4·H2The concentration of O is from about 0.13mg/mL to about 1.90 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, said Na2HPO4·12H2The concentration of O is from about 2.25mg/mL to about 4.12mg/mL, the NaH2PO4·H2The concentration of O is from about 1.16mg/mL to about 1.90 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, said Na2HPO4·12H2The concentration of O is about 4.12mg/mL, the NaH2PO4·H2The concentration of O was about 1.16 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application has an osmolality of no more than 600 mOsmoL/kg.
In certain embodiments, the pharmaceutical formulation of the present application has an osmolality between about 270mOsmoL/kg and about 590 mOsmoL/kg.
In certain embodiments, the pharmaceutical formulation of the present application has an osmolality between about 270mOsmoL/kg and about 400 mOsmoL/kg.
In certain embodiments, the pharmaceutical formulation of the present application has an osmolality between about 270mOsmoL/kg and about 300 mOsmoL/kg.
In certain embodiments, the pharmaceutical formulation of the present application has an osmotic pressure selected from the group consisting of: about 276, about 284, about 362, about 372, about 417, about 432, about 453, about 459, about 498, about 516, about 535, about 586, and about 588 mOsmoL/kg.
In certain embodiments, the pharmaceutical formulation of the present application has an osmotic pressure selected from the group consisting of: about 276mOsmol/kg, and about 284 mOsmol/kg.
In certain embodiments, the pharmaceutical formulation of the present application has an osmolality of about 276 mOsmoL/kg.
In certain embodiments, the pharmaceutical formulation of the present application further comprises a stabilizer.
In certain embodiments, the pharmaceutical formulation of the present application, the stabilizing agent is selected from the group consisting of: mannitol and sodium chloride.
In certain embodiments, the pharmaceutical formulation of the present application, the mannitol is at a concentration of about 0mg/mL to about 50 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the mannitol is at a concentration of about 15mg/mL to about 45 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the mannitol is at a concentration of about 15mg/mL to about 30 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the mannitol is at a concentration of about 0mg/mL, about 15mg/mL, about 30mg/mL, about 45mg/mL, or about 50 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the mannitol is at a concentration of about 30 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of sodium chloride is from about 2mg/mL to about 12 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of sodium chloride is from about 2mg/mL to about 10 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of sodium chloride is from about 2mg/mL to about 9.5 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of sodium chloride is from about 2mg/mL to about 7.5 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of sodium chloride is from about 2mg/mL to about 5 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of sodium chloride is about 2mg/mL, about 5mg/mL, about 7.5mg/mL, about 9.5mg/mL, about 10mg/mL, or about 12 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, wherein the sodium chloride is at a concentration of about 2 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application further comprises a surfactant.
In certain embodiments, the pharmaceutical formulation of the present application, the surfactant is selected from the group consisting of: poloxamer 188, and tween 20.
In certain embodiments, the pharmaceutical formulation of the present application, the concentration of poloxamer 188 is from about 0mg/mL to about 0.1 mg/mL.
In certain embodiments, the pharmaceutical formulation of the present application, the tween 20 is at a concentration of from about 0mg/mL to about 0.2 mg/mL.
In certain embodiments, a pharmaceutical formulation of the present application comprises methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, about 20mM buffer, mannitol at a concentration of about 0mg/mL to about-50 mg/mL, and sodium chloride at a concentration of about 2mg/mL to about-12 mg/mL, at a pH of between about 6.5 and about 7.5.
In certain embodiments, a pharmaceutical formulation of the present application comprises about 0.1mg/mL methionine, about 0.9mg/mL thyrotropin, about 20mM buffer having a pH of about 7.0, about 30mg/mL mannitol, and about 2mg/mL sodium chloride.
In certain embodiments, a pharmaceutical formulation of the present application comprises methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, Na at about 4.12mg/mL2HPO4·12H2O, NaH at about 1.16mg/mL2PO4·H2O, about 30mg/mL mannitol, and about 2mg/mL sodium chloride.
In certain embodiments, the pharmaceutical formulation of the present application, in a dosage form comprising an injectable formulation.
In certain embodiments, the pharmaceutical formulation of the present application, in a dosage form comprising a liquid injection.
In certain embodiments, the pharmaceutical formulation of the present application, in a dosage form comprising an injection of water.
In another aspect, the present application provides a kit comprising a pharmaceutical formulation of the present application.
In another aspect, the present application provides a method of preparing a pharmaceutical formulation of the present application comprising adding methionine to thyrotropin, adding thyrotropin to methionine and/or adding methionine and thyrotropin simultaneously.
In certain embodiments, the addition of a buffer having a pH of about 7.0, mannitol, sodium chloride, and water is also included.
In certain embodiments, the pharmaceutical formulation comprises an injectable formulation.
In certain embodiments, the methods of the present application further comprise filling the pharmaceutical formulation into a pre-filled needle or an injection vial.
In certain embodiments, the pharmaceutical formulation is filled into an injection vial in a range of about 0.5mL to about 2.0mL per vial.
In certain embodiments, the pharmaceutical formulation is filled in an injection vial at about 1.2 mL/vial.
In certain embodiments, the methods of the present application further comprise sealing the injection vial.
In certain embodiments, the pharmaceutical formulation is filled into the pre-filled needle at about 0.1mL to about 2.0mL per branch.
In certain embodiments, the pharmaceutical formulation is filled into the pre-filled needle at about 0.5mL to about 2.0mL per branch.
In certain embodiments, the pharmaceutical formulation is filled into the pre-filled needle at about 0.5mL to about 1.0mL per branch.
In certain embodiments, the pharmaceutical formulation is filled into the pre-filled needle at about 0.5 mL/branch, about 1.0 mL/branch, or about 2.0 mL/branch.
In certain embodiments, the pharmaceutical formulation is filled into the pre-filled needle at about 1.0 mL/branch.
In another aspect, the present application provides a method of affecting thyroxine levels comprising administering to a subject a pharmaceutical formulation of the present application.
In certain embodiments, the methods of the present application, wherein affecting the thyroxine level comprises increasing and/or maintaining the affected thyroxine level.
In another aspect, the present application provides a method of affecting the thyroid comprising administering to a subject a pharmaceutical formulation of the present application.
In certain embodiments, the affecting the thyroid comprises affecting the volume of the thyroid, affecting the function of the thyroid, and/or affecting the blood supply to the thyroid.
In certain embodiments, the function of the thyroid comprises thyroid synthesis and/or secretion of thyroxine.
In another aspect, the present application provides a method of affecting thyroid cells comprising administering to a subject a pharmaceutical formulation of the present application.
In certain embodiments, the affecting the thyroid cell comprises affecting proliferation of the thyroid cell and/or affecting a function of the thyroid cell.
In certain embodiments, the function of the thyroid cell comprises synthesis and/or secretion of thyroxine by the thyroid cell.
In another aspect, the present application provides a use of a pharmaceutical formulation as described herein in the manufacture of a medicament for the diagnosis, assisted diagnosis, prevention, assisted treatment and/or treatment of a thyroid disorder in a subject.
In certain embodiments, the thyroid disease comprises autoimmune thyroid disease and/or thyroid cancer.
In certain embodiments, the thyroid disorder comprises hypothyroidism.
In certain embodiments, the thyroid disorder comprises a psychotic disorder associated with hypothyroidism.
In certain embodiments, the thyroid disorder comprises goiter.
In certain embodiments, the thyroid disorder comprises hashimoto's disease.
In certain embodiments, the thyroid disease comprises Differentiated Thyroid Carcinoma (DTC).
Other aspects and advantages of the present application will be readily apparent to those skilled in the art from the following detailed description. Only exemplary embodiments of the present application have been shown and described in the following detailed description. As those skilled in the art will recognize, the disclosure of the present application enables those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention as it is directed to the present application. Accordingly, the descriptions in the drawings and the specification of the present application are illustrative only and not limiting.
Drawings
The specific features of the invention to which this application relates are set forth in the appended claims. The features and advantages of the invention to which this application relates will be better understood by reference to the exemplary embodiments described in detail below and the accompanying drawings. The brief description of the drawings is as follows:
FIG. 1 shows the results of the subunit oxidation product content in thyrotropin-like samples at day 0 and day 14 at 40. + -. 2 ℃ for 8 formulations of the first trial of the present application.
FIG. 2 shows the results of SE-HPLC analysis of major peak content in thyrotrophin-like samples at day 0, 14 and 28 at elevated temperature 40. + -. 2 ℃ for 8 prescriptions of test one of the present application.
FIG. 3 shows the results of SE-HPLC analysis of the content of the dimer in the thyrotropin-like samples at day 0, 14 and 28 at elevated temperature 40. + -. 2 ℃ in 8 prescriptions of test one of the present application.
FIG. 4 shows the results of SE-HPLC analysis of the degradation levels of thyrotropin-like samples at day 0, 14 and 28 at elevated temperatures of 40. + -. 2 ℃ in 8 formulations of the first trial of the present application.
FIG. 5 shows the detection of thyrotropin protein levels in 8 prescription samples of test one of the present applications, day 0 and day 14 thyrotropin samples at elevated temperature 40. + -. 2 ℃.
FIG. 6 shows the results of measuring the subunit oxidation product content of thyrotropin-like substances at day 0, day 30 and day 60 at 25. + -. 2 ℃ for 12 prescriptions of test two of the present application.
FIG. 7 shows the results of SE-HPLC analysis of the main peak content in thyrotrophin-like samples at day 0, day 30 and day 60 at 25. + -. 2 ℃ for 12 prescriptions in test two of the present application.
FIG. 8 shows the results of SE-HPLC analysis of the amount of dimer in the thyrotropin-like samples at day 0, day 30 and day 60 at 25. + -. 2 ℃ for 12 prescriptions in test two of the present application.
FIG. 9 shows the results of SE-HPLC analysis of the degradation levels of thyrotropin-like substances at day 0, day 30 and day 60 at 25. + -. 2 ℃ in 12 prescriptions of test two of the present application.
FIG. 10 shows the results of measuring the subunit oxidation product content of thyrotropin-like substances at day 0, day 30 and day 60 at 25. + -. 2 ℃ for 3 prescriptions of test III of the present application.
FIG. 11 shows the results of SE-HPLC analysis of the major peak content in thyrotropin-like samples at day 0, day 30 and day 60 at 25. + -. 2 ℃ for 3 prescriptions of test three of the present application.
FIG. 12 shows the results of SE-HPLC analysis of the dimer content of thyrotropin-like samples at day 0, day 30 and day 60 at 25. + -. 2 ℃ for 3 prescriptions of trial three of the present application.
FIG. 13 shows the results of measuring the subunit oxidation product content of thyrotropin-like substances at day 0 and day 30 at 40. + -. 2 ℃ for samples of formulas 1, 2, 3 and 4 of test four of the present application.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification.
Definition of terms
In the present application, the term "expression" generally refers to transcription and/or translation occurring within a host cell. The level of transcription of a desired product in a host cell can be determined based on the amount of the corresponding mRNA present in the cell. For example, mRNA transcribed from a selected sequence can be quantified by PCR or Northern hybridization (see, Sambrook et al, molecular cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York (1989)). The protein encoded by the selected sequence may be quantified by a variety of methods, e.g., by ELISA, by detection of biological activity of the protein, or by using assays independent of such activity, e.g., Western blotting or radioimmunoassay (using antibodies that recognize and bind the protein in the assay) (see, supra, Sambrook et al, 1989). For example, the thyrotropin of the present application may be expressed in mammalian cells, for example, the thyrotropin of the present application may be expressed in mouse cells, for example, the thyrotropin of the present application may be expressed in Chinese Hamster Ovary (CHO) cells.
In the present application, the term "surfactant" generally refers to an organic substance having an amphoteric structure. They may consist of groups of opposite solubility tendencies, and may consist of an oil-soluble hydrocarbon chain and a water-soluble ionic group. Surfactants can be classified into anionic, cationic and nonionic surfactants according to the charge of the surface active moiety. Surfactants can be used as wetting, emulsifying, solubilizing and dispersing agents for various pharmaceutical compositions and biomaterial formulations. For example, the surfactant herein can comprise an ionic surfactant and/or a nonionic surfactant, e.g., the surfactant herein can comprise a polyoxyethylene-type, a polyol-type, an alkanolamide-type, a polyether-type, and/or an amine oxide-type surfactant, e.g., the surfactant herein can comprise poloxamer 188 and/or tween 20.
In the present application, the terms "Thyrotropin" and "Thyroid stimulating hormone" are used interchangeably and "Thyrotropin" (TSH or Thyrotropin) in the present application generally refers to a peptide hormone secreted by Thyroid stimulating cells in the anterior pituitary. For example, the thyroid stimulating hormone of the present application may include human thyrotropin (hTSH). hTSH can be a macromolecular glycoprotein secreted by the human pituitary, which contains about 210 amino acids and can consist of an alpha chain and a beta chain. The alpha chain may be common to both thyrotropin and gonadotropin, while the beta chain may be characteristic of thyrotropin. The human endogenous thyrotropin action site can be thyroid cells and thyroid cancer cells containing thyrotropin receptors. The physiological actions of thyrotropin may include various links to promote synthesis and release of thyrotropin, enhance sodium/iodine co-transporter (NIS) and peroxidase activities of thyroid cells, increase thyroglobulin (Tg) synthesis and/or tyrosine iodination; thyrotropin promotes metabolism and intracellular nucleic acid and protein synthesis in thyroid epithelial cells, resulting in cell proliferation and/or enlargement of the gland. In the present application, the term "Recombinant Human thyrotropin" (rhTSH) generally refers to a Human thyrotropin obtained by introducing a nucleotide sequence comprising Human thyrotropin into a host cell and expressing the same. For example, the recombinant human thyrotropin may comprise a recombinant human thyrotropin alpha subunit (the GeneID encoding it may be 7252), a recombinant human thyrotropin beta subunit, or a combination thereof. For example, the recombinant human thyrotropin of the present application may comprise recombinant human thyrotropin described in us patent 5,840,566. For example, the recombinant human thyrotropin of the present application may comprise recombinant human thyrotropin described in us patent 5,674,711. For example, the thyrotropins of the present application may comprise various homologues of thyrotropins as well as thyrotropins from various species. For example, a thyrotropin herein may comprise a functionally active fragment of thyrotropin, e.g., a thyrotropin α subunit (e.g., NP _000726.1), a thyrotropin β subunit (e.g., NP _000540.2), or a combination thereof.
In the present application, the term "buffer" generally refers to an agent that allows a liquid formulation to resist changes in pH. In certain embodiments, the added buffer allows the liquid formulation to resist pH changes through the action of its acid-base conjugated components. For example, the buffer of the present application may comprise a phosphate buffer.
In the present application, the term "phosphate buffer" generally refers to a buffer comprising sodium dihydrogen phosphate and/or disodium hydrogen phosphate or hydrates thereof.
In the present application, the term "methionine" generally refers to α -amino- γ -methylthiobutanoic acid. For example, a methionine of the present application may comprise the D configuration and/or the L configuration.
In the present application, the term "thyroid" generally refers to an endocrine organ. The thyroid is usually located below the thyroid cartilage in the neck, on either side of the trachea. The thyroid gland may have the effect of controlling the rate of energy use, producing proteins, regulating the body's sensitivity to other hormones. The thyroid gland can produce thyroxine, regulate metabolism and growth rate, and regulate other body systems. The thyroid gland also produces Calcitonin (Calcitonin), which regulates the balance of calcium in the body.
In this application, the term "function of the thyroid gland" or "thyroid function" generally refers to the role played by the thyroid gland in an animal. For example, thyroid function of the present application may comprise synthesis, secretion and/or release of thyroxine. For example, thyroid function of the present application may comprise synthesis, secretion and/or release of thyroxine.
In this application, the term "thyroxine levels" generally refers to the amount of thyroxine in an animal. For example, the thyroxine levels of the present application can be the intracellular and/or extracellular concentration of thyroxine. For example, the thyroxine levels of the present application can be the concentration of thyroxine within and/or outside of a tissue. For example, the thyroxine levels of the present application may be the concentration of thyroxine in the serum.
In the present application, the term "poloxamer" generally refers to a nonionic triblock copolymer composed of a polypropylene oxide (PPO) central hydrophobic chain and two polyethylene oxide (PEO) hydrophilic chains flanking, each PPO or PEO chain may have a different molecular weight. For example, the poloxamer can be poloxamer 188, a poloxamer having PPO chains with a molecular weight of 1800g/mol and a PEO content of 80% (w/w) of the poloxamer.
In the present application, the term "osmotic pressure" generally refers to a measure of the osmotic pressure of dissolved solute particles in an aqueous solution. Solute particles may include ionic and non-ionic molecules. The osmolality can be expressed as the concentration of osmotically active particles (i.e., osmolality) dissolved in 1kg of solution (1 mOsm/kg H at 38 ℃; 1mOsm/kg H2O corresponds to an osmotic pressure of 19mm Hg). In this application, the abbreviation "mOsm" generally refers to "milliosmoles per kg of solution".
In the present application, the term "pH" generally refers to the hydrogen ion concentration index in a liquid, and may refer to, for example, the ratio of the total number of hydrogen ions in a solution to the amount of total substances. In the present application, the pH may be determined by a pH indicator, a pH paper, and/or a pH meter.
In the present application, the term "biological activity" generally refers to the ability of a molecule to affect any physical or biochemical property, pathway, molecule, or interaction of a biological system associated with an organism (including but not limited to viruses, bacteria, phages, transposons, prions, insects, fungi, plants, animals, and humans). For example, biological activity herein may refer to thyrotropin promoting growth and development, affecting metabolic rate, and/or affecting neurological function. For example, the biological activity of thyrotropin of the present application may be affected by the content of thyrotropin degradation products, oxidation products and/or aggregates, e.g. an increase in thyrotropin degradation products, oxidation products and/or aggregates may result in a decrease in the biological activity of thyrotropin.
In the present application, the term "administering" generally refers to introducing a pharmaceutical formulation of the present application into the body of a subject by any route of introduction or delivery. Any method known to those skilled in the art for contacting a cell, organ or tissue with a pharmaceutical combination of the present application may be employed. Administration of the present application may include, without limitation, intravenous, intra-arterial, intranasal, intra-abdominal, intravenous, intramuscular, subcutaneous transdermal or oral administration. Administration of the present application may include, without limitation, hip intramuscular injection. A daily dose may be divided into one, two or more doses of suitable form to be administered at one, two or more times during a certain period of time.
In the present application, the term "kit" generally refers to two or more components packaged together in a container, receptacle, or other container, which contains a pharmaceutical formulation of the present application. For example, a kit can comprise a pharmaceutical formulation of the present application.
In the present application, the term "subject" generally refers to an animal, which may generally be a mammal, such as a human, a non-human primate (ape, gibbon, gorilla, chimpanzee, orangutan, macaque), a domestic animal (dog and cat), a farm animal (poultry such as chickens and ducks, horses, cattle, goats, sheep, pigs) and a laboratory animal (mouse, rat, rabbit, guinea pig). Human subjects may include fetal, neonatal, infant, juvenile and adult subjects. The subject can include animal disease models, such as mouse models, and other animal models known to those of skill in the art.
In the present application, the term "tween" refers generally to polysorbate surfactants, such as polyoxyethylene derivatives of sorbitan monolaurate, for example tween 20 may be polysorbate 20 (polyoxyethylene (20) sorbitan monolaurate)Cinnamate). The number 20 after the polyoxyethylene may refer to the oxyethylene- (CH) s found in the molecule2CH2O) -total number of groups. The numbers following "polysorbate" may refer to the type of fatty acid associated with the polyoxyethylene sorbitan portion of the molecule. For example, monolaurate may be represented by 20, monopalmitate may be represented by 40, monostearate may be represented by 60, and monooleate may be represented by 80. For example, tween 20 of the present application may be substituted by another detergent or surfactant, such as tween 40, tween 60 and/or tween 80.
In the present application, the term "stabilizer" generally refers to a pharmaceutical excipient that protects the active pharmaceutical ingredient and/or formulation from stability during manufacture, storage and/or use. Stabilizers may include, but are not limited to, salts and/or polyols. For example, the stabilizer may include sodium chloride and/or mannitol.
In the present application, the term "pharmaceutical formulation" generally refers to a composition which is in such a form as to allow the biological activity of the active ingredient to be effective and which does not contain additional components which are unacceptably toxic to the subject to whom the formulation will be administered. The invention provides a pharmaceutical preparation, which contains recombinant human thyrotropin and a pharmaceutically acceptable carrier or excipient.
In this application, the term "injectable" refers generally to solutions, emulsions and/or suspensions intended for injection into the body, as well as sterile powders or concentrated solutions intended for constitution as a solution or suspension immediately prior to use.
In the present application, the term "liquid injection" or "injectable liquid injection" generally refers to a solution, emulsion and/or suspension for injection into the body. For example, a liquid injection may be a preparation prepared by formulating a drug into a solution, emulsion and/or suspension, and filling it into an injection bottle or a multi-dose container.
In the present application, the term "liquid injection" or "injectable liquid injection" generally refers to a solution for injection into the body. For example, an injection solution may be prepared by formulating a drug into a solution and filling the solution into a vial or a multi-dose container.
In the present application, the term "injection vial" generally refers to any container filled with a medicament. For example, the injection bottle of the present application may be a glass bottle. For example, the injection bottle of the present application may be an ampoule bottle. For example, the injection bottle of the present application may be an ampoule bottle filled with the pharmaceutical preparation of the present application.
In the present application, the term "pre-filled needle" generally refers to a pre-filled syringe that is filled with a medicament. For example, the pre-filled needle of the present application may be a syringe filled with the pharmaceutical formulation of the present application.
In the present application, the term "prevention" generally refers to the prevention of the recurrence, onset, or development of one or more symptoms of a disorder in an individual. For example, it may be to prevent recurrence, onset or progression of thyroid disease.
In the present application, the term "adjuvant therapy" is generally used to enhance the primary therapeutic effect and/or to restore health. Restoring health of the present application may comprise restoring volume, function, and/or blood supply to an organ (and/or tissue) of the subject. The adjunctive therapy of the present application may be performed before, after, and/or simultaneously with the primary therapy. The primary treatment of the present application may include, but is not limited to, surgical treatment, drug treatment, and/or radiation treatment. For example, the adjuvant therapy can be a therapy that can be used to enhance the primary therapeutic effect and/or restore the health of the subject before and after the primary treatment of the thyroid disorder using the pharmaceutical formulation of the present application. For example, the pharmaceutical formulations of the present application can be used for restoring thyroid function in a subject, which can include, but is not limited to, before and after surgical treatment for thyroid cancer.
In this application, "aided diagnosis" generally refers to a method that can be helpful in making a clinical determination regarding the presence or nature of a particular type of symptom or condition. For example, the pharmaceutical formulations of the present application may be used in the assisted diagnosis of thyroid disorders.
In the present application, the term "diagnosing" generally refers to detecting a disease or condition, or determining the state or extent of a disease or condition. The term "diagnosing" also includes detecting a predisposition for a disease or condition, determining the therapeutic effect of a drug treatment, or predicting a response pattern to a drug treatment or xenobiotic. For example, the status or extent of thyroid disease may be detected, or determined.
In this application, "parts by weight" or "parts by weight" are used interchangeably and can be any fixed weight expressed in micrograms, milligrams, grams, or kilograms (e.g., 1 ug, 1mg, 1g, 2g, 5g, or kg, etc.). For example, a composition consisting of 1 part by weight of component a and 9 parts by weight of component b may be a composition consisting of 1g of component a +9 g of component b, or 10 g of component a +90 g of component b. In the compositions of the present application, the percentage content of a certain component is (parts by weight of that component/sum of parts by weight of all components) x 100%. Thus, in a composition consisting of 1 part by weight of component a and 9 parts by weight of component b, the content of component a is 10% and component b is 90%.
In addition, the above "parts by weight" may be converted into "number of moles" in the solution state; the "weight ratio" may also be converted to a "molar concentration ratio". The weight unit of the weight ratio of the application can be as follows: any weight unit such as kilogram (kg), milligram (mg), microgram (μ g); the molar units of the molar (concentration) ratio may be: molarity (M), millimolar (mM), micromolar (. mu.M) and the like.
In the present application, the term "comprising" is generally intended to include the explicitly specified features, but not to exclude other elements.
In the present application, the term "about" generally refers to a variation in the range of 0.5% -10% above or below the specified value, such as a variation in the range of about 0.5%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 5.5%, about 6%, about 6.5%, about 7%, about 7.5%, about 8%, about 8.5%, about 9%, about 9.5%, or about 10% above or below the specified value.
Detailed Description
In one aspect, the present application provides a pharmaceutical formulation comprising thyrotropin and methionine. In one embodiment, the methionine of the present application may comprise L-methionine. In one embodiment, the concentration of methionine herein may be 0.005mg/mL or more. In one embodiment, the methionine of the present application may be at a concentration of at least about 0.001mg/mL, at least about 0.002mg/mL, at least about 0.003mg/mL, at least about 0.004mg/mL, at least about 0.005mg/mL, at least about 0.01mg/mL, at least about 0.02mg/mL, at least about 0.03mg/mL, at least about 0.04mg/mL, at least about 0.05mg/mL, at least about 0.1mg/mL, at least about 0.15mg/mL, at least about 0.2mg/mL, at least about 0.3mg/mL, at least about 0.4mg/mL, at least about 0.5mg/mL, at least about 1mg/mL, at least about 1.5mg/mL, at least about 2mg/mL, at least about 2.5mg/mL, at least about 3mg/mL, at least about 3.5mg/mL, at least about 3mg/mL, At least about 4mg/mL, at least about 4.5mg/mL, at least about 5mg/mL, at least about 5.5mg/mL, at least about 6mg/mL, at least about 6.5mg/mL, at least about 7mg/mL, at least about 7.5mg/mL, at least about 8mg/mL, at least about 8.5mg/mL, at least about 9mg/mL, at least about 9.5mg/mL, at least about 10mg/mL, at least about 11mg/mL, at least about 12mg/mL, at least about 13mg/mL, at least about 14mg/mL, at least about 15mg/mL, at least about 16mg/mL, at least about 17mg/mL, at least about 18mg/mL, at least about 19mg/mL, at least about 20mg/mL, at least about 21mg/mL, at least about 6mg/mL, At least about 22mg/mL, at least about 23mg/mL, at least about 24mg/mL, at least about 25mg/mL, at least about 30mg/mL, at least about 35mg/mL, at least about 40mg/mL, at least about 45mg/mL, at least about 50mg/mL, at least about 55mg/mL, at least about 60mg/mL, at least about 65mg/mL, at least about 70mg/mL, at least about 75mg/mL, at least about 80mg/mL, at least about 85mg/mL, at least about 90mg/mL, at least about 95mg/mL, at least about 100mg/mL, or more.
In one embodiment, the concentration of methionine herein may be from about 0.005mg/mL to about 1.5 mg/mL. In one embodiment, the methionine of the present application may be at a concentration of about 0.001mg/mL to about 10mg/mL, about 0.002mg/mL to about 10mg/mL, about 0.003mg/mL to about 10mg/mL, about 0.004mg/mL to about 10mg/mL, about 0.005mg/mL to about 10mg/mL, about 0.01mg/mL to about 10mg/mL, about 0.001mg/mL to about 0.2mg/mL, about 0.002mg/mL to about 0.2mg/mL, about 0.003mg/mL to about 0.2mg/mL, about 0.004mg/mL to about 0.2mg/mL, about 0.005mg/mL to about 0.2mg/mL, about 0.01mg/mL to about 0.2mg/mL, about 0.02mg/mL to about 0.2mg/mL, About 0.03mg/mL to about 0.2mg/mL, about 0.04mg/mL to about 0.2mg/mL, about 0.05mg/mL to about 0.2mg/mL, about 0.02mg/mL to about 10mg/mL, about 0.03mg/mL to about 10mg/mL, about 0.04mg/mL to about 10mg/mL, about 0.05mg/mL to about 10mg/mL, about 0.1mg/mL to about 10mg/mL, about 0.15mg/mL to about 10mg/mL, about 0.2mg/mL to about 10mg/mL, about 0.3mg/mL to about 10mg/mL, about 0.4mg/mL to about 10mg/mL, about 0.5mg/mL to about 10mg/mL, about 1mg/mL to about 10mg/mL, about 1.5mg/mL to about 10mg/mL, About 2mg/mL to about 10mg/mL, about 2.5mg/mL to about 10mg/mL, about 3mg/mL to about 10mg/mL, about 3.5mg/mL to about 10mg/mL, about 4mg/mL to about 10mg/mL, about 4.5mg/mL to about 10mg/mL, about 5mg/mL to about 10mg/mL, about 5.5mg/mL to about 10mg/mL, about 6mg/mL to about 10mg/mL, about 6.5mg/mL to about 10mg/mL, about 7mg/mL to about 10mg/mL, about 7.5mg/mL to about 10mg/mL, about 8mg/mL to about 10mg/mL, about 8.5mg/mL to about 10mg/mL, about 9mg/mL to about 10mg/mL, or, About 9.5mg/mL to about 10mg/mL, or about 9.9mg/mL to about 10 mg/mL. In one embodiment, the methionine of the present application may be present in a concentration of about 0.001mg/mL to about 2mg/mL, about 0.002mg/mL to about 2mg/mL, about 0.003mg/mL to about 2mg/mL, about 0.004mg/mL to about 2mg/mL, about 0.005mg/mL to about 2mg/mL, about 0.01mg/mL to about 2mg/mL, about 0.02mg/mL to about 2mg/mL, about 0.03mg/mL to about 2mg/mL, about 0.04mg/mL to about 2mg/mL, about 0.05mg/mL to about 2mg/mL, about 0.1mg/mL to about 2mg/mL, about 0.15mg/mL to about 2mg/mL, about 0.2mg/mL to about 2mg/mL, or, About 0.3mg/mL to about 2mg/mL, about 0.4mg/mL to about 2mg/mL, about 0.5mg/mL to about 2mg/mL, about 1mg/mL to about 2mg/mL, or about 1.5mg/mL to about 2 mg/mL. In one embodiment, the methionine of the present application may be present in a concentration of about 0.001mg/mL to about 1.5mg/mL, about 0.002mg/mL to about 1.5mg/mL, about 0.003mg/mL to about 1.5mg/mL, about 0.004mg/mL to about 1.5mg/mL, about 0.005mg/mL to about 1.5mg/mL, about 0.01mg/mL to about 1.5mg/mL, about 0.02mg/mL to about 1.5mg/mL, about 0.03mg/mL to about 1.5mg/mL, about 0.04mg/mL to about 1.5mg/mL, about 0.05mg/mL to about 1.5mg/mL, about 0.1mg/mL to about 1.5mg/mL, about 0.15mg/mL to about 1.5mg/mL, about 0.2mg/mL to about 1.5mg/mL, About 0.3mg/mL to about 1.5mg/mL, about 0.4mg/mL to about 1.5mg/mL, about 0.5mg/mL to about 1.5mg/mL, or about 1mg/mL to about 1.5 mg/mL. In one embodiment, the methionine of the present application may be at a concentration of about 0.001mg/mL to about 0.2mg/mL, about 0.002mg/mL to about 0.2mg/mL, about 0.003mg/mL to about 0.2mg/mL, about 0.004mg/mL to about 0.2mg/mL, about 0.005mg/mL to about 0.2mg/mL, about 0.01mg/mL to about 0.2mg/mL, about 0.02mg/mL to about 0.2mg/mL, about 0.03mg/mL to about 0.2mg/mL, about 0.04mg/mL to about 0.2mg/mL, about 0.05mg/mL to about 0.2mg/mL, about 0.1mg/mL to about 0.2mg/mL, or about 0.15mg/mL to about 0.2 mg/mL. In one embodiment, the concentration of methionine herein may be from about 0.001mg/mL to about 0.1mg/mL, from about 0.002mg/mL to about 0.1mg/mL, from about 0.003mg/mL to about 0.1mg/mL, from about 0.004mg/mL to about 0.1mg/mL, from about 0.005mg/mL to about 0.1mg/mL, from about 0.01mg/mL to about 0.1mg/mL, from about 0.02mg/mL to about 0.1mg/mL, from about 0.03mg/mL to about 0.1mg/mL, from about 0.04mg/mL to about 0.1mg/mL, or from about 0.05mg/mL to about 0.1 mg/mL. In one embodiment, the methionine of the present application may be present in a concentration of about 0.1mg/mL to about 0.15mg/mL, about 0.1mg/mL to about 0.2mg/mL, about 0.1mg/mL to about 0.3mg/mL, about 0.1mg/mL to about 0.4mg/mL, about 0.1mg/mL to about 0.5mg/mL, about 0.1mg/mL to about 1mg/mL, about 0.1mg/mL to about 1.5mg/mL, about 0.1mg/mL to about 2mg/mL, about 0.1mg/mL to about 2.5mg/mL, about 0.1mg/mL to about 3mg/mL, about 0.1mg/mL to about 3.5mg/mL, about 0.1mg/mL to about 4mg/mL, about 0.1mg/mL to about 4.5mg/mL, about 0.1mg/mL to about 5mg/mL, About 0.1mg/mL to 5.5mg/mL, about 0.1mg/mL to 6mg/mL, about 0.1mg/mL to 6.5mg/mL, about 0.1mg/mL to 7mg/mL, about 0.1mg/mL to 7.5mg/mL, about 0.1mg/mL to 8mg/mL, about 0.1mg/mL to 8.5mg/mL, about 0.1mg/mL to 9mg/mL, about 0.1mg/mL to 9.5mg/mL, or about 0.1mg/mL to 10 mg/mL.
In one embodiment, the thyrotropin of the present application may be selected from the group consisting of: thyrotropin isolated from mammals, and recombinant mammalian thyrotropin. In one embodiment, the thyrotropin of the present application may comprise recombinant human thyrotropin. In one embodiment, the thyrotropin of the present application may comprise recombinant human thyrotropin expressed by a cell. In one embodiment, the thyrotropin of the present application may comprise a recombinant human thyrotropin expressed by mammalian cells. In one embodiment, the thyrotropin of the present application may comprise recombinant human thyrotropin expressed by mouse cells. In one embodiment, the thyrotropin of the present application may comprise recombinant human thyrotropin expressed by Chinese Hamster Ovary (CHO) cells. In one embodiment, the thyrotropin of the present application may comprise recombinant human thyrotropin expressed by transfected Chinese Hamster Ovary (CHO) cells. In one embodiment, the thyrotropin of the present application may comprise recombinant human thyrotropin expressed by Chinese Hamster Ovary (CHO) cells after transfection of a recombinant human thyrotropin expression vector. In one embodiment, the amino acid sequence of thyrotropin of the present application is described in the literature. In one embodiment, the amino acid sequence of thyrotropin of the present application may comprise recombinant human thyrotropin as described in U.S. patent No. 5,840,566. In one embodiment, the amino acid sequence of thyrotropin of the present application may comprise recombinant human thyrotropin as described in U.S. patent No. 5,674,711. In one embodiment, the amino acid sequence of thyrotropin of the present application may comprise the group: recombinant human thyrotropin alpha subunits, recombinant human thyrotropin beta subunits, and combinations thereof. The working reference substance of the recombinant human thyrotropin can be prepared according to Chinese pharmacopoeia, generally refers to a batch which is stable enough and suitable for clinical tests, the working reference substance in the application can be calibrated according to the Chinese pharmacopoeia on protein concentration, purity and biological activity, and the biological activity can be calibrated by comparing the Chinese pharmacopoeia with thyrotropin of WHO international standard. For example, the international standard for thyrotropin may be selected from the group consisting of: NIBSC reference 00/504, NIBSC reference 81/565, NIBSC reference 94/674, and NIBSC reference 03/192.
In one embodiment, the concentration of thyrotropin herein may be from about 0.45mg/mL to about 1.8 mg/mL. In one embodiment, the concentration of thyrotropin may be at least about 0.01mg/mL, at least about 0.02mg/mL, at least about 0.03mg/mL, at least about 0.04mg/mL, at least about 0.05mg/mL, at least about 0.1mg/mL, at least about 0.15mg/mL, at least about 0.2mg/mL, at least about 0.3mg/mL, at least about 0.4mg/mL, at least about 0.5mg/mL, at least about 0.6mg/mL, at least about 0.7mg/mL, at least about 0.8mg/mL, at least about 0.9mg/mL, at least about 1mg/mL, at least about 1.1mg/mL, at least about 1.2mg/mL, at least about 1.3mg/mL, at least about 1.4mg/mL, at least about 1.5mg/mL, at least about 2mg/mL, At least about 2.5mg/mL, at least about 3mg/mL, at least about 3.5mg/mL, at least about 4mg/mL, at least about 4.5mg/mL, at least about 5mg/mL, at least about 5.5mg/mL, at least about 6mg/mL, at least about 6.5mg/mL, at least about 7mg/mL, at least about 7.5mg/mL, at least about 8mg/mL, at least about 8.5mg/mL, at least about 9mg/mL, at least about 9.5mg/mL, at least about 10mg/mL, or more.
In one embodiment, the concentration of thyrotropin of the present application may be from about 0.01mg/mL to about 5mg/mL, from about 0.02mg/mL to about 5mg/mL, from about 0.03mg/mL to about 5mg/mL, from about 0.04mg/mL to about 5mg/mL, from about 0.05mg/mL to about 5mg/mL, from about 0.1mg/mL to about 5mg/mL, from about 0.15mg/mL to about 5mg/mL, from about 0.2mg/mL to about 5mg/mL, from about 0.3mg/mL to about 5mg/mL, from about 0.4mg/mL to about 5mg/mL, from about 0.5mg/mL to about 5mg/mL, from about 0.6mg/mL to about 5mg/mL, from about 0.7mg/mL to about 5mg/mL, or, About 0.8mg/mL to about 5mg/mL, about 0.9mg/mL to about 5mg/mL, about 1mg/mL to about 5mg/mL, about 1.1mg/mL to about 5mg/mL, about 1.2mg/mL to about 5mg/mL, about 1.3mg/mL to about 5mg/mL, about 1.4mg/mL to about 5mg/mL, about 1.5mg/mL to about 5mg/mL, about 2mg/mL to about 5mg/mL, about 2.5mg/mL to about 5mg/mL, about 3mg/mL to about 5mg/mL, about 3.5mg/mL to about 5mg/mL, about 4mg/mL to about 5mg/mL, or about 4.5mg/mL to about 5 mg/mL. In one embodiment, the concentration of thyrotropin of the present application may be from about 0.01mg/mL to 1.8mg/mL, from about 0.02mg/mL to 1.8mg/mL, from about 0.03mg/mL to 1.8mg/mL, from about 0.04mg/mL to 1.8mg/mL, from about 0.05mg/mL to 1.8mg/mL, from about 0.1mg/mL to 1.8mg/mL, from about 0.15mg/mL to 1.8mg/mL, from about 0.2mg/mL to 1.8mg/mL, from about 0.3mg/mL to 1.8mg/mL, from about 0.4mg/mL to 1.8mg/mL, from about 0.5mg/mL to 1.8mg/mL, from about 0.6mg/mL to 1.8mg/mL, from about 0.7mg/mL to 1.8mg/mL, from about 0.8mg/mL to 1.8mg/mL, About 0.9mg/mL to 1.8mg/mL, about 1mg/mL to 1.8mg/mL, about 1.1mg/mL to 1.8mg/mL, about 1.2mg/mL to 1.8mg/mL, about 1.3mg/mL to 1.8mg/mL, about 1.4mg/mL to 1.8mg/mL, or about 1.5mg/mL to 1.8 mg/mL. In one embodiment, the concentration of thyrotropin of the present application may be from about 0.01mg/mL to about 0.9mg/mL, from about 0.02mg/mL to about 0.9mg/mL, from about 0.03mg/mL to about 0.9mg/mL, from about 0.04mg/mL to about 0.9mg/mL, from about 0.05mg/mL to about 0.9mg/mL, from about 0.1mg/mL to about 0.9mg/mL, from about 0.15mg/mL to about 0.9mg/mL, from about 0.2mg/mL to about 0.9mg/mL, from about 0.3mg/mL to about 0.9mg/mL, from about 0.4mg/mL to about 0.9mg/mL, from about 0.5mg/mL to about 0.9mg/mL, from about 0.6mg/mL to about 0.9mg/mL, from about 0.7mg/mL to about 0.9mg/mL, from about 0.8mg/mL to about 0.9mg/mL, About 0.81mg/mL to 0.9mg/mL, about 0.82mg/mL to 0.9mg/mL, about 0.83mg/mL to 0.9mg/mL, about 0.84mg/mL to 0.9mg/mL, about 0.85mg/mL to 0.9mg/mL, about 0.86mg/mL to 0.9mg/mL, about 0.87mg/mL to 0.9mg/mL, about 0.88mg/mL to 0.9mg/mL, or about 0.89mg/mL to 0.9 mg/mL. In one embodiment, the concentration of thyrotropin of the present application may be from about 0.9mg/mL to about 0.91mg/mL, from about 0.9mg/mL to about 0.92mg/mL, from about 0.9mg/mL to about 0.93mg/mL, from about 0.9mg/mL to about 0.91mg/mL, from about 0.9mg/mL to about 0.95mg/mL, from about 0.9mg/mL to about 0.96mg/mL, from about 0.9mg/mL to about 0.97mg/mL, from about 0.9mg/mL to about 0.98mg/mL, from about 0.9mg/mL to about 0.99mg/mL, from about 0.9mg/mL to about 1mg/mL, from about 0.9mg/mL to about 1.1mg/mL, from about 0.9mg/mL to about 1.2mg/mL, from about 0.9mg/mL to about 1.3mg/mL, About 0.9mg/mL to about 1.4mg/mL, about 0.9mg/mL to about 1.5mg/mL, about 0.9mg/mL to about 2mg/mL, about 0.9mg/mL to about 2.5mg/mL, about 0.9mg/mL to about 3mg/mL, about 0.9mg/mL to about 3.5mg/mL, about 0.9mg/mL to about 4mg/mL, or about 0.9mg/mL to about 4.5 mg/mL.
In another aspect, the pH of the pharmaceutical formulation of the present application can be between about 6.5 and about 7.5. In one embodiment, the pH of the pharmaceutical formulation of the present application may be about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.1, about 7.2, about 7.3, about 7.4, or about 7.5. In one embodiment, the pH of the pharmaceutical formulation of the present application may be between about 6.5 and about 7.5, between about 6.6 and about 7.5, between about 6.7 and about 7.5, between about 6.8 and about 7.5, between about 6.9 and about 7.5, between about 7.0 and about 7.5, between about 7.1 and about 7.5, between about 7.2 and about 7.5, between about 7.3 and about 7.5, between about 7.4 and about 7.5, or between about 7.45 and about 7.5. In one embodiment, the pH of the pharmaceutical formulation of the present application may be between about 6.5 and about 7.0, between about 6.6 and about 7.0, between about 6.7 and about 7.0, between about 6.8 and about 7.0, between about 6.9 and about 7.0, or between about 6.95 and about 7.0.
In another aspect, the pharmaceutical formulation of the present application may comprise a buffering agent. In one embodiment, the buffer of the present application may comprise a phosphate buffer. In one embodiment, the buffer of the present application can be a phosphate buffer between about 6.5 and about 7.5, between about 6.6 and about 7.5, between about 6.7 and about 7.5, between about 6.8 and about 7.5, between about 6.9 and about 7.5, between about 7.0 and about 7.5, between about 7.1 and about 7.5, between about 7.2 and about 7.5, between about 7.3 and about 7.5, between about 7.4 and about 7.5, or between about 7.45 and about 7.5. In one embodiment, the buffer of the present application can be a phosphate buffer between about 6.5 and about 7.0, between about 6.6 and about 7.0, between about 6.7 and about 7.0, between about 6.8 and about 7.0, between about 6.9 and about 7.0, or between about 6.95 and about 7.0. In one embodiment, the buffer of the present application may be a phosphate buffer of about 7.0. In one embodiment, the buffer of the present application may be at a concentration of about 20 mM.
In one embodiment, the buffer of the present application may comprise the group of: na (Na)2HPO4·12H2O, and NaH2PO4·H2And O. Na of the present application2HPO4·12H2The concentration of O can be from about 2.25mg/mL to about 6.82mg/mL, NaH, of the present application2PO4·H2The concentration of O may be from about 0.13mg/mL to about 1.90 mg/mL. In one embodiment, Na of the present application2HPO4·12H2The concentration of O can be from about 2.25mg/mL to about 4.12mg/mL, NaH of the present application2PO4·H2The concentration of O may be from about 1.16mg/mL to about 1.90 mg/mL. Na in this application when the buffer is at about pH6.52HPO4·12H2The concentration of O may be about 2.25mg/mL, NaH, of the present application2PO4·H2The concentration of O may be about 1.90 mg/mL. Na in this application when the buffer is at about pH7.02HPO4·12H2The concentration of O, NaH of the present application, may be about 4.12mg/mL2PO4·H2The concentration of O may be about 1.16 mg/mL. Na of the present application when the buffer is at about pH7.52HPO4·12H2The concentration of O may be about 6.82mg/mL, NaH, of the present application2PO4·H2The concentration of O may be about 0.13 mg/mL.
In another aspect, the osmotic pressure of the pharmaceutical formulation of the present application may not exceed 600 mOsmol/kg. In one embodiment, the osmolality of the pharmaceutical formulation of the present application can be no more than 600mOsmoL/kg, no more than 590mOsmoL/kg, no more than 588mOsmoL/kg, no more than 586mOsmoL/kg, no more than 535mOsmoL/kg, no more than 516mOsmoL/kg, no more than 498mOsmoL/kg, no more than 459mOsmoL/kg, no more than 453mOsmoL/kg, no more than 432mOsmoL/kg, no more than 417mOsmoL/kg, no more than 372mOsmoL/kg, no more than 362mOsmoL/kg, no more than 284mOsmoL/kg, no more than 276mOsmoL/kg, or less. In one embodiment, the osmotic pressure of a pharmaceutical formulation of the present application can be between about 270mOsmoL/kg and about 590 mOsmoL/kg. In one embodiment, the osmotic pressure of a pharmaceutical formulation of the present application can be between about 270mOsmoL/kg and about 400 mOsmoL/kg. In one embodiment, the osmotic pressure of a pharmaceutical formulation of the present application can be between about 270mOsmoL/kg and about 300 mOsmoL/kg. In one embodiment, the osmotic pressure of the pharmaceutical formulation of the present application may be selected from the group consisting of: about 276mOsmol/kg, about 284mOsmol/kg, about 362mOsmol/kg, about 372mOsmol/kg, about 417mOsmol/kg, about 432mOsmol/kg, about 453mOsmol/kg, about 459mOsmol/kg, about 498mOsmol/kg, about 516mOsmol/kg, about 535mOsmol/kg, about 586mOsmol/kg, and about 588 mOsmol/kg.
In another aspect, the pharmaceutical formulations of the present application may comprise a stabilizer. In one embodiment, the stabilizer of the present application may be selected from the group consisting of: mannitol and sodium chloride. In one embodiment, the mannitol of the present application may be at a concentration of about 0mg/mL to about 30mg/mL, about 15mg/mL to about 30mg/mL, about 30mg/mL to about 45mg/mL, or about 30mg/mL to about 50 mg/mL. In one embodiment, the concentration of mannitol herein may be from about 0mg/mL to about 50mg/mL, from about 15mg/mL to about 45mg/mL, or from about 15mg/mL to about 30 mg/mL. In one embodiment, the concentration of mannitol herein may be about 0mg/mL, about 15mg/mL, about 30mg/mL, about 45mg/mL, or about 50 mg/mL. In one embodiment, the mannitol of the present application can be at a concentration of about 2mg/mL to about 12mg/mL, about 2mg/mL to about 10mg/mL, about 2mg/mL to about 9.5mg/mL, about 2mg/mL to about 7.5mg/mL, or about 2mg/mL to about 5 mg/mL. In one embodiment, the concentration of mannitol herein may be about 2mg/mL, about 5mg/mL, about 7.5mg/mL, about 9.5mg/mL, about 10mg/mL, or about 12 mg/mL.
In another aspect, the pharmaceutical formulation of the present application may comprise a surfactant. In one embodiment, the surfactant of the present application may be selected from the group consisting of: poloxamer, and tween. In one embodiment, the poloxamers of the present application may comprise poloxamer 188. In one embodiment, the concentration of poloxamer 188 of the present application is between about 0mg/mL and about 0.1 mg/mL. In one embodiment, the tween herein may comprise tween 20. In one embodiment, the concentration of tween 20 herein is from about 0mg/mL to about 0.2 mg/mL.
In another aspect, a pharmaceutical formulation of the present application may comprise methionine at a concentration of from about 0.005mg/mL to about-1.5 mg/mL, and thyrotropin at a concentration of from about 0.45mg/mL to about-1.8 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, and about 20mM buffer, pH between 6.5 and 7.5. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, and mannitol at a concentration of about 0mg/mL to about-50 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, and sodium chloride at a concentration of about 2mg/mL to about-12 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, about 20mM buffer, and sodium chloride at a concentration of about 2mg/mL to about-12 mg/mL, at a pH of between 6.5 and 7.5. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, about 20mM buffer, and mannitol at a concentration of about 0mg/mL to about-50 mg/mL, at a pH of between 6.5 and 7.5. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, mannitol at a concentration of about 0mg/mL to about-50 mg/mL, and sodium chloride at a concentration of about 2mg/mL to about-12 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at a concentration of between about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of between about 0.45mg/mL to about-1.8 mg/mL, about 20mM buffer, mannitol at a concentration of between about 0mg/mL to about-50 mg/mL, and sodium chloride at a concentration of between about 2mg/mL to about-12 mg/mL, at a pH of between 6.5 and 7.5.
In another aspect, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, and thyrotropin at about 0.9 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, and a buffer at about 20mM at a pH of about 7.0. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, and mannitol at about 30 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, and sodium chloride at about 2 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, mannitol at about 30mg/mL, and sodium chloride at about 2 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise about 0.1mg/mL methionine, about 0.9mg/mL thyrotropin, about 20mM buffer having a pH of about 7.0, and about 2mg/mL sodium chloride. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, a buffer at about 20mM at a pH of about 7.0, and mannitol at about 30 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, a buffer at about 20mM having a pH of about 7.0, mannitol at about 30mg/mL, and sodium chloride at about 2 mg/mL. In one embodiment, a pharmaceutical formulation of the present application may comprise methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, Na at about 4.12mg/mL2HPO4·12H2O, NaH at about 1.16mg/mL2PO4·H2O, about 30mg/mL mannitol and about2mg/mL sodium chloride, the osmolality may be 276 mOsmol/kg.
In another aspect, the pharmaceutical formulation of the present application may comprise an injectable formulation. In one embodiment, a pharmaceutical formulation of the present application may comprise a liquid injection. The pharmaceutical preparation of the application can contain injection solution.
The present application also provides a kit. In one embodiment, a kit of the present application may comprise a pharmaceutical formulation of the present application and optionally instructions for use.
The application also provides a preparation method. In one embodiment, the methods of the present disclosure may include adding methionine to thyrotropin, adding thyrotropin to methionine, and/or adding methionine and thyrotropin simultaneously. In one embodiment, the methods of the present application can include adding methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, and thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL. In one embodiment, the method of the present application may further comprise adding a buffer having a pH of about 7.0, mannitol, sodium chloride, and optionally water. In one embodiment, the methods of the present application may comprise adding methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, about 20mM buffer, mannitol at a concentration of about 0mg/mL to about-50 mg/mL, and sodium chloride at a concentration of about 2mg/mL to about-12 mg/mL, at a pH of between 6.5 and 7.5, and optionally water.
In another aspect, the methods of preparation provided herein can comprise filling a pharmaceutical formulation of the present application in a container. In one embodiment, the pharmaceutical formulation of the present application may be filled into a pre-filled needle or vial. In one embodiment, a pharmaceutical formulation of the present application may be filled into a pre-filled needle. In one embodiment, the pharmaceutical formulation of the present application may be filled into an injection vial. In one embodiment, the pharmaceutical formulation of the present application may be filled into vials. In one embodiment, sealing the container of the present application may also be included. In one embodiment, the preparation method provided herein can fill the pharmaceutical formulation of the present application into an injection vial in an amount of about 0.5mL to about 2.0mL per vial. In one embodiment, the pharmaceutical formulation of the present application can be filled into an injection vial in an amount of from about 0.5mL to about 1.2mL per vial. In one embodiment, the pharmaceutical formulation of the present application can be filled into an injection vial in an amount of from about 1.2mL to about 2.0mL per vial. In one embodiment, a vial of a pharmaceutical formulation of the present application may be filled at least about 0.5mL, at least about 1.0mL, at least about 1.2mL, or at least about 2.0mL per vial. In one embodiment, the pharmaceutical formulation of the present application can be filled into a pre-filled needle at about 0.1mL to about 2.0mL per branch. In one embodiment, the pharmaceutical formulation of the present application can be filled into a pre-filled needle at about 0.5mL to about 2.0mL per arm, 0.5mL to about 1.0mL per arm, or about 1.0mL to about 2.0mL per arm. In one embodiment, a pre-filled needle can be filled with a pharmaceutical formulation of the present application at least about 0.5mL/, at least about 1.0 mL/branch, or at least about 2.0 mL/branch. For example, when the concentration of thyrotropin in the drug formulation of the present application is about 0.45mg/mL, the drug formulation of the present application can be filled into the pre-filled needle at least about 2.0 mL/branch. For example, when the concentration of thyrotropin in the drug formulation of the present application is about 0.9mg/mL, the drug formulation of the present application can be filled into the pre-filled needle at least about 1.0 mL/branch. For example, when the concentration of thyrotropin in the drug formulation of the present application is at least about 1.35mg/mL, the drug formulation of the present application can be filled into the pre-filled needle at about 0.67 mL/branch. For example, when the concentration of thyrotropin in the drug formulation of the present application is at least about 1.8mg/mL, the drug formulation of the present application can be filled into the pre-filled needle at about 0.5 mL/branch.
In another aspect, the present application also provides a method of affecting thyroxine levels. In one embodiment, the methods provided herein for affecting thyroxine levels can be in vitro. For example, the pharmaceutical formulations of the present application can be administered to cells and/or tissues in vitro, and can be used to affect thyroxine levels in cells and/or tissues in vitro. In one embodiment, the methods provided herein for affecting thyroxine levels can be ex vivo. For example, the pharmaceutical formulations of the present application can be administered to cells and/or tissues ex vivo and can be used to affect the thyroxine levels in cells and/or tissues ex vivo, or in subjects who have received such ex vivo cells and/or tissues after introduction of the ex vivo cells and/or tissues into the subject. In one embodiment, the methods provided herein for affecting thyroxine levels can be of non-diagnostic interest. In one embodiment, the methods provided herein for affecting thyroxine levels may be of non-therapeutic interest. In one embodiment, the methods provided herein for affecting thyroxine levels can be of non-diagnostic and non-therapeutic interest. In one embodiment, the methods provided herein for affecting thyroxine levels can comprise increasing and/or maintaining the affected thyroxine levels. For example, it may involve affecting the metabolism of the thyroid gland. For example, promoting synthesis and release of thyroxine, enhancing the activity of sodium/iodine co-transporter (NIS) and peroxidase of the thyroid, increasing thyroglobulin (Tg) synthesis and/or iodination of tyrosine may be included.
In another aspect, the present application also provides a method of affecting the thyroid. In one embodiment, a method of affecting the thyroid provided herein can comprise affecting the volume of the thyroid, affecting the function of the thyroid, and/or affecting the blood supply to the thyroid. In one embodiment, a method of affecting the thyroid provided herein can comprise affecting thyroid synthesis and/or secretion of thyroxine.
In another aspect, the present application also provides a method of affecting a thyroid cell. In one embodiment, the methods provided herein for affecting thyroid cells can be in vitro. In one embodiment, the methods provided herein for affecting thyroid cells can be ex vivo. In one embodiment, the methods provided herein for affecting thyroid cells can be of non-diagnostic interest. In one embodiment, the methods provided herein for affecting thyroid cells can be of non-therapeutic interest. In one embodiment, the methods provided herein for affecting thyroid cells can be for non-diagnostic and non-therapeutic purposes. In one embodiment, the methods provided herein for affecting thyroid cells can comprise affecting proliferation of thyroid cells and/or affecting function of thyroid cells. In one embodiment, the methods provided herein for affecting a thyroid cell can comprise affecting a thyroid cell to synthesize and/or secrete thyroxine. For example, it may involve affecting the metabolism of thyroid cells. For example, promoting synthesis and release of thyroxine, enhancing sodium/iodine co-transporter (NIS) and peroxidase activity in thyroid cells, increasing thyroglobulin (Tg) synthesis and/or iodination of tyrosine may be included.
In another aspect, the present application also provides a use of a pharmaceutical formulation of the present application in the manufacture of a medicament for the diagnosis, assisted diagnosis, prevention, assisted treatment and/or treatment of a thyroid disorder in a subject. In one embodiment, the thyroid disorders of the present application can include autoimmune thyroid disorders and/or thyroid cancer. The thyroid disorders of the present application may be selected from the group consisting of: hypothyroidism, mental disorders associated with hypothyroidism, goiter, and hashimoto's disease. In one embodiment, the thyroid disorder herein can comprise Differentiated Thyroid Cancer (DTC). In one embodiment, the present application further provides a use of the pharmaceutical formulation of the present application for the manufacture of a medicament for the adjunctive diagnosis of serum thyroglobulin (Tg) during follow-up after thyroidectomy in patients with Differentiated Thyroid Carcinoma (DTC) and/or for the adjunctive treatment with radioiodine to remove residual thyroid tissue after peri-or peri-thyroidectomy in patients without distant metastasis Differentiated Thyroid Carcinoma (DTC).
In another aspect, the present application also provides a method of diagnosing, aiding in diagnosing, preventing, aiding in treating, and/or treating a thyroid disorder. In one embodiment, the thyroid disorders of the present application can include autoimmune thyroid disorders and/or thyroid cancer. The thyroid disorders of the present application may be selected from the group consisting of: hypothyroidism, mental disorders associated with hypothyroidism, goiter, and hashimoto's disease. In one embodiment, the thyroid disorder herein can comprise Differentiated Thyroid Cancer (DTC). In one embodiment, the present application further provides a method for the auxiliary diagnosis of serum thyroglobulin (Tg) during follow-up after thyroidectomy in patients with Differentiated Thyroid Cancer (DTC) and/or a method for the auxiliary treatment of patients with metastatic differentiated thyroid cancer without distal metastasis by removing residual thyroid tissue with radioactive iodine after peri-or total thyroidectomy.
In another aspect, the present application also provides a pharmaceutical formulation of the present application, which can be used for diagnosis, assisted diagnosis, prevention, assisted treatment and/or treatment of thyroid disorders. In another aspect, the present application also provides a pharmaceutical formulation of the present application for use in the diagnosis, co-diagnosis, prevention and/or treatment of autoimmune thyroid disease and/or thyroid cancer. In another aspect, the present application also provides a pharmaceutical formulation of the present application, which can be used for the diagnosis, auxiliary diagnosis, prevention and/or treatment of thyroid disorders, which thyroid disorders of the present application can be selected from the group consisting of: hypothyroidism, mental disorders associated with hypothyroidism, goiter, and hashimoto's disease. In another aspect, the present application also provides a pharmaceutical formulation of the present application that can be used for the diagnosis, co-diagnosis, prevention and/or treatment of thyroid disorders, which can include Differentiated Thyroid Cancer (DTC). In another aspect, the present application also provides a pharmaceutical formulation of the present application, which can be used for the assisted diagnosis of serum thyroglobulin (Tg) during the follow-up after thyroidectomy in patients with Differentiated Thyroid Cancer (DTC) and/or for the assisted treatment of patients with Differentiated Thyroid Cancer (DTC) without distant metastasis with radioiodine to remove residual thyroid tissue after peri-or total thyroidectomy.
Technical scheme
1. A pharmaceutical formulation comprising thyrotropin and methionine, wherein the methionine is at a concentration of about 0.005mg/mL or greater.
2. The pharmaceutical formulation of claim 1, wherein the methionine comprises L-methionine.
3. The pharmaceutical formulation of any one of claims 1-2, wherein the methionine is at a concentration of about 0.005mg/mL to about 1.5 mg/mL.
4. The pharmaceutical formulation of any one of claims 1-3, wherein the methionine is at a concentration of about 0.02mg/mL to about 1.5 mg/mL.
5. The pharmaceutical formulation of any one of claims 1-3, wherein the methionine is at a concentration of about 0.005mg/mL, about 0.02mg/mL, about 0.1mg/mL, about 0.2mg/mL, or about 1.5 mg/mL.
6. The pharmaceutical formulation of any one of claims 1-5, wherein the methionine is at a concentration of about 0.1 mg/mL.
7. The pharmaceutical formulation according to any of claims 1-6, wherein the thyrotropin is selected from the group consisting of: thyrotropin isolated from mammals, and recombinant mammalian thyrotropin.
8. The pharmaceutical formulation of any one of claims 1-7, wherein the thyrotropin comprises a cell-expressed recombinant human thyrotropin.
9. The pharmaceutical formulation of any one of claims 1-8, wherein the thyrotropin comprises a mammalian cell-expressed recombinant human thyrotropin.
10. The pharmaceutical formulation of any one of claims 1-9, wherein the thyrotropin comprises recombinant human thyrotropin expressed by Chinese Hamster Ovary (CHO) cells.
11. The pharmaceutical formulation of any one of claims 1-10, wherein the concentration of thyrotropin is from about 0.45mg/mL to about 1.8 mg/mL.
12. The pharmaceutical formulation of any one of claims 1-11, wherein the concentration of thyrotropin is from about 0.9mg/mL to about 1.8 mg/mL.
13. The pharmaceutical formulation of any one of claims 1-11, wherein the concentration of thyrotropin is about 0.45mg/mL, about 0.9mg/mL, or about 1.8 mg/mL.
14. The pharmaceutical formulation of any one of claims 1-13, wherein the concentration of thyrotropin is about 0.9 mg/mL.
15. The pharmaceutical formulation of any one of claims 1-14, having a pH between about 6.5 and about 7.5.
16. The pharmaceutical formulation of any one of claims 1-15, having a pH between about 6.5 and about 7.0.
17. The pharmaceutical formulation of any one of claims 1-16, having a pH of about 6.5 or about 7.0.
18. The pharmaceutical formulation of any one of claims 1-17, having a pH of about 7.0.
19. The pharmaceutical formulation of any one of claims 1-18, further comprising a buffering agent.
20. The pharmaceutical formulation of claim 19, wherein the buffer comprises a phosphate buffer.
21. The pharmaceutical formulation of any one of claims 19-20, wherein the buffer comprises a phosphate buffer having a pH between about 6.5 and about 7.5.
22. The pharmaceutical formulation of any one of claims 19-21, wherein the buffer is at a concentration of about 20 mM.
23. The pharmaceutical formulation of any one of claims 19-22, wherein the buffer has a pH of about 7.0.
24. The pharmaceutical formulation according to any one of claims 19-23, wherein the buffering agent comprises the group of: na (Na)2HPO4·12H2O, and NaH2PO4·H2O。
25. The pharmaceutical formulation of claim 24, wherein the Na2HPO4·12H2The concentration of O is from about 2.25mg/mL to about 6.82 mg/mL.
26. The pharmaceutical formulation of any one of claims 24-25, the NaH2PO4·H2The concentration of O is from about 0.13mg/mL to about 1.90 mg/mL.
27. The pharmaceutical formulation of claim 26, the Na2HPO4·12H2The concentration of O is from about 2.25mg/mL to about 4.12mg/mL, the NaH2PO4·H2The concentration of O is from about 1.16mg/mL to about 1.90 mg/mL.
28. The pharmaceutical formulation of any one of claims 26-27, said Na2HPO4·12H2The concentration of O is about 4.12mg/mL, the NaH2PO4·H2The concentration of O was about 1.16 mg/mL.
29. The pharmaceutical formulation of any one of claims 1-28, having an osmolality of no more than 600 mOsmoL/kg.
30. The pharmaceutical formulation of any one of claims 1-29, having an osmolality between about 270mOsmoL/kg and about 590 mOsmoL/kg.
31. The pharmaceutical formulation of any one of claims 1-30, having an osmolality between about 270mOsmoL/kg and about 400 mOsmoL/kg.
32. The pharmaceutical formulation of any one of claims 1-31, having an osmolality between about 270mOsmoL/kg and about 300 mOsmoL/kg.
33. The pharmaceutical formulation according to any of claims 1-30, having an osmotic pressure selected from the group consisting of: about 276, about 284, about 362, about 372, about 417, about 432, about 453, about 459, about 498, about 516, about 535, about 586, and about 588 mOsmoL/kg.
34. The pharmaceutical formulation according to any of claims 1-33, having an osmotic pressure selected from the group consisting of: about 276mOsmol/kg, and about 284 mOsmol/kg.
35. The pharmaceutical formulation of any one of claims 1-34, having an osmolality of about 276 mOsmoL/kg.
36. The pharmaceutical formulation of any one of claims 1-35, further comprising a stabilizer.
37. The pharmaceutical formulation of claim 36, wherein the stabilizer is selected from the group consisting of: mannitol and sodium chloride.
38. The pharmaceutical formulation of claim 37, wherein the mannitol is at a concentration of about 0mg/mL to about 50 mg/mL.
39. The pharmaceutical formulation of any one of claims 37-38, wherein the mannitol is at a concentration of about 15mg/mL to about 45 mg/mL.
40. The pharmaceutical formulation of any one of claims 37-39, wherein the mannitol is at a concentration of about 15mg/mL to about 30 mg/mL.
41. The pharmaceutical formulation of any one of claims 37-38, wherein the mannitol is at a concentration of about 0mg/mL, about 15
mg/mL, about 30mg/mL, about 45mg/mL, or about 50 mg/mL.
42. The pharmaceutical formulation of any one of claims 37-41, wherein the mannitol is at a concentration of about 30 mg/mL.
43. The pharmaceutical formulation of any one of claims 37-42, wherein the sodium chloride is at a concentration of about 2mg/mL to about 12 mg/mL.
44. The pharmaceutical formulation of any one of claims 37-43, wherein the sodium chloride is at a concentration of about 2mg/mL to about 10 mg/mL.
45. The pharmaceutical formulation of any one of claims 37-44, wherein the sodium chloride is at a concentration of about 2mg/mL to about 9.5 mg/mL.
46. The pharmaceutical formulation of any one of claims 37-45, wherein the sodium chloride is at a concentration of about 2mg/mL to about 7.5 mg/mL.
47. The pharmaceutical formulation of any one of claims 37-46, wherein the sodium chloride is at a concentration of about 2mg/mL to about 5 mg/mL.
48. The pharmaceutical formulation of any one of claims 37-43, wherein the sodium chloride is at a concentration of about 2mg/mL, about 5mg/mL, about 7.5mg/mL, about 9.5mg/mL, about 10mg/mL, or about 12 mg/mL.
49. The pharmaceutical formulation of any one of claims 37-48, wherein the sodium chloride is at a concentration of about 2 mg/mL.
50. The pharmaceutical formulation of any one of claims 1-49, further comprising a surfactant.
51. The pharmaceutical formulation of claim 50, wherein the surfactant is selected from the group consisting of: poloxamer 188, and tween 20.
52. The pharmaceutical formulation of claim 51, wherein the concentration of poloxamer 188 is between about 0mg/mL and about 0.1 mg/mL.
53. The pharmaceutical formulation of claims 51-52, wherein the tween 20 is at a concentration of about 0mg/mL to about 0.2 mg/mL.
54. The pharmaceutical formulation of any one of claims 1-53, comprising methionine at a concentration of about 0.005mg/mL to about-1.5 mg/mL, thyrotropin at a concentration of about 0.45mg/mL to about-1.8 mg/mL, about 20mM buffer, mannitol at a concentration of about 0mg/mL to about-50 mg/mL, and sodium chloride at a concentration of about 2mg/mL to about-12 mg/mL, at a pH of between about 6.5 and about 7.5.
55. The pharmaceutical formulation of any one of claims 1-54, comprising about 0.1mg/mL methionine, about 0.9mg/mL thyrotropin, about 20mM buffer having a pH of about 7.0, about 30mg/mL mannitol, and about 2mg/mL sodium chloride.
56. The pharmaceutical formulation of any one of claims 1-55, comprising methionine at about 0.1mg/mL, thyrotropin at about 0.9mg/mL, Na at about 4.12mg/mL2HPO4·12H2O, NaH at about 1.16mg/mL2PO4·H2O, about 30mg/mL mannitol, and about 2mg/mL sodium chloride.
57. The pharmaceutical formulation of any one of claims 1-56, wherein the dosage form comprises an injectable formulation.
58. The pharmaceutical preparation according to any one of claims 1 to 57, wherein the dosage form comprises a liquid injection.
59. The pharmaceutical preparation according to any one of claims 1 to 58, wherein the dosage form comprises a water injection.
60. A kit comprising the pharmaceutical formulation of any one of claims 1-59.
61. A method of making a pharmaceutical formulation according to any of claims 1-59, comprising adding methionine to thyrotropin, adding thyrotropin to methionine and/or adding methionine and thyrotropin simultaneously.
62. The method of claim 61, further comprising adding a buffer having a pH of about 7.0, mannitol, sodium chloride, and water.
63. The method of any of claims 61-62, wherein the pharmaceutical formulation comprises an injectable formulation.
64. The method of any of claims 61-63, further comprising filling the pharmaceutical formulation into a pre-filled needle or an injection vial.
65. The method of claim 64, wherein the pharmaceutical formulation is filled into the vial of injection at a rate of about 0.5mL to about 2.0mL per vial.
66. The method of any of claims 64-65, filling the pharmaceutical formulation into an injection vial at about 1.2 mL/vial.
67. The method of any of claims 64-66, further comprising sealing the injection vial.
68. According to the method of claim 64, filling the pharmaceutical formulation into the pre-filled needle in an amount of about 0.1mL to about 2.0mL per branch.
69. Filling the pharmaceutical formulation into the pre-filled needle at about 0.5mL to about 2.0mL per branch according to the method of claim 64 or 68.
70. According to the method of any of claims 64 and 68-69, filling the pharmaceutical formulation into the pre-filled needle at about 0.5mL to about 1.0mL per branch.
71. According to the method of any of claims 64 and 68-70, the pharmaceutical formulation is filled into the pre-filled needle at about 0.5 mL/arm, about 1.0 mL/arm, or about 2.0 mL/arm.
72. According to the method of any of claims 64 and 68-71, the pharmaceutical formulation is filled into the pre-filled needle at about 1.0 mL/branch.
73. A method of affecting thyroxine levels comprising administering to a subject a pharmaceutical formulation of any of claims 1-59.
74. The method of claim 73, wherein affecting the thyroxine level comprises increasing and/or maintaining the affected thyroxine level.
75. A method of affecting the thyroid comprising administering to a subject a pharmaceutical formulation of any one of claims 1-59.
76. The method of claim 75, wherein affecting the thyroid comprises affecting thyroid volume, affecting thyroid function, and/or affecting thyroid blood supply.
77. The method of any one of claims 75-76, wherein the function of the thyroid comprises thyroid synthesis and/or thyroxine secretion.
78. A method of affecting thyroid cells comprising administering to a subject a pharmaceutical formulation of any one of claims 1-59.
79. The method of claim 78, wherein affecting the thyroid cell comprises affecting proliferation of the thyroid cell and/or affecting a function of the thyroid cell.
80. The method of any of claims 78-79, wherein the function of the thyroid cell comprises synthesis and/or secretion of thyroxine by the thyroid cell.
81. Use of a pharmaceutical formulation of any one of claims 1-59 in the manufacture of a medicament for diagnosing, aiding diagnosis, preventing, aiding treatment, and/or treating a thyroid disorder in a subject.
82. The use according to claim 81, wherein the thyroid disease comprises autoimmune thyroid disease and/or thyroid cancer.
83. The use according to any one of claims 81-82, wherein the thyroid disorder comprises hypothyroidism.
84. The use according to any of claims 81-83, wherein the thyroid disorder comprises a psychotic disorder associated with hypothyroidism.
85. The use according to any one of claims 81-84, wherein the thyroid disorder comprises goiter.
86. The use according to any one of claims 81-85, wherein the thyroid disorder comprises Hashimoto's disease.
87. The use according to any one of claims 81-86, wherein the thyroid disease comprises Differentiated Thyroid Cancer (DTC).
Without intending to be bound by any theory, the following examples are merely intended to illustrate the fusion proteins, preparation methods, uses, etc. of the present application, and are not intended to limit the scope of the invention of the present application.
Examples
Example 1
1.1 materials and instruments
The reagent information and the instrument information used in the examples of the present application are shown in tables 1 and 2 below, respectively.
TABLE 1 reagent information
| Name of reagent | Manufacturer of the product | Goods number |
| Disodium hydrogen phosphate dodecahydrate | Sigma | 04273-1kg |
| Monohydrate sodium dihydrogen phosphate | Merck | 106349 9025 |
| Sodium chloride | Merck | 137017 5000 |
| Mannitol | Sigma | M8429-500G |
| Poloxamer 188 | Merck | 1.37065.1000 |
| L-methionine | Sigma | M5308-100g |
| Tween 20 | Merck | 817061 1000 |
TABLE 2 Instrument information
| Name of instrument | Manufacturer of the product | Model number |
| Nucleic acid protein detector | Thermo | Nanodrop 2000 |
| Desk type centrifuge | Beckman Coulter | AllegraTM X-12R-Centrifuge |
| Ultra-low temperature refrigerator at minus 80 DEG C | Haier | DW-87L 386 |
| Drug stability experiment box | SHANGHAI YIHENG INSTR Co.,Ltd. | TEMI880 |
| Enzyme-linked immunosorbent assay (ELISA) instrument | Molecular Devices | Spectramax 384plus |
| PH meter | EUTECH | pH700 |
| Balance with a movable handle | Shanghai Precision Scientific Instrument Co., Ltd. | JY2002 |
| Chromatograph | Shimadzu | LC-20AT |
1.2 items of examination
And (3) detecting the content of subunit oxidation products: the content of subunit oxidation products is determined by referring to the high performance liquid chromatography 0512 of the three general rules of China pharmacopoeia 2015 edition or the high performance liquid chromatography commonly used in the field.
And (3) detection by SE-HPLC: SE-HPLC (molecular exclusion chromatography) is a liquid chromatography technology for separating components with different molecular weights, different retention times in a chromatographic column, a peak first of a component with a larger molecular weight and a peak later of a component with a smaller molecular weight according to the molecular weight of the component to be detected, and refers to 0514 molecular exclusion chromatography in the three general rules of Chinese pharmacopoeia 2015 edition or molecular exclusion chromatography commonly used in the field.
And (3) protein content detection: referring to the SE-HPLC protein assay, the calculation formula for the protein sample content at day n may be: cn＝(A2×C0×V1)/(A1×V2). Wherein A is1Represents the total peak area of the protein sample at day 0, A2Represents the total peak area, V, of the protein sample at the n-th day of the test group1Denotes the initial sample volume on day 0, V2Represents the protein sample injection volume at day 0 of the test group, C0Represents the protein sample content at day 0, CnRepresents the content of protein sample at the nth day of the test group.
And (3) biological activity detection: adding samples with final concentration of 0.0128-5000ng/mL into 96-well plate, and spreading cells at 2.5 × 1 per well04The TSHR-HEK293 cell stably expressing the human TSH receptor is reacted at 37 ℃ for 30min, then the cell is cracked, and the supernatant is taken to carry out cAMP ELISA detection, and the light absorption value is read at 450nm by taking 570nm as a reference wavelength. The data can be fitted with four parameters by SoftMax, and the S-shaped curve of the working reference product is used as a contrast to perform parallel fitting on the curve of the sample to obtain the EC of each detection product50And calculating the relative biological activity of the sample.
Preparation of thyrotropin: the methods described in thyrotropin references (e.g., U.S. patent 5,674,711) may be recombinant human thyrotropin expressed in Chinese Hamster Ovary (CHO) cells; the thyrotropin preparation method is fully described in examples 1-6 of U.S. patent 5,674,711, and the recombinant human thyrotropin prepared by known methods can be used in the pharmaceutical formulations of the present application.
1.3 run one: effect of different substances on the storage Effect of liquid formulations
This example explores the effect of different substances on the storage effect of a thyrotropin liquid formulation. The prescription composition for each test group of trial one is shown in table 3. The liquid formulations were subjected to accelerated stability testing at 40 ± 2 ℃ for 4 weeks to simulate the effect of long term storage under extreme conditions on the samples.
Table 3 prescription composition for each test group of test one
Note: the protein concentration represents the protein concentration of recombinant human thyrotropin, and the PB buffer represents phosphate buffer.
Subunit oxidation product content detection result and analysis
As shown in FIG. 1, using the 8 prescription samples from test one above, results of the subunit oxidation product content in thyrotrophin-like samples at day 0 and day 14 at high temperature of 40. + -. 2 ℃ were obtained. The results showed that the subunit oxidation product content of formulas 1, 2, 3, 7 and 8 of trial one, without methionine addition or with other substances addition, increased after 14 days at an elevated temperature of 40 ± 2 ℃; whereas formulations 4, 5 and 6 of trial one, with methionine added, produced almost no subunit oxidation products.
SE-HPLC DETECTION AND ANALYSIS
As shown in FIG. 2, the results of measuring the content of the main peak in the thyrotrophin-like substance at day 0, 14 and 28 at a high temperature of 40. + -. 2 ℃ by SE-HPLC using 8 prescribed samples of the first test described above. As shown in FIG. 3, using 8 prescribed samples of the first test, the results of measuring the content of the dimer in the thyrotrophin-like substance at day 0, 14 and 28 at high temperature of 40. + -. 2 ℃ by SE-HPLC were obtained. As shown in FIG. 4, the results of measuring the degradation content of thyroid stimulating hormone samples at 0 th, 14 th and 28 th days at a high temperature of 40. + -. 2 ℃ by SE-HPLC using 8 prescribed samples of the first test. The results show that the addition of methionine to test one formulations 4, 5 and 6 reduced the production of polymer products compared to test one formulations 1, 2 and 3 without methionine added, which did not significantly affect the content of degradation products.
Protein content detection results and analysis
As shown in FIG. 5, the thyrotropin content was measured in thyrotropin-like samples at day 0 and day 14 at high temperature 40. + -. 2 ℃ using 8 prescriptions of the first test described above. The results show that after 14 days at an elevated temperature of 40 ± 2 ℃, the thyrotropin content of formulas 4, 5 and 6 of trial one is not significantly reduced and the addition of methionine may facilitate storage of the thyrotropin liquid formulations.
Thus, the addition of methionine may facilitate storage of the thyrotropin liquid formulation, and may retain the active ingredient of the thyrotropin liquid formulation, as compared to other protein component stabilizing materials such as poloxamer 188 or tween 20.
Example 2
And (2) test II: effect of different sodium chloride and mannitol concentrations on the storage Effect of liquid formulations
This example explores the effect of different sodium chloride and mannitol concentrations on the storage effect of thyrotropin liquid formulations. The prescription composition of each test group for test two is shown in table 4. The liquid formulation was subjected to accelerated stability testing at 25 ± 2 ℃ for 60 days to simulate the effect of long term storage conditions under extreme conditions on the sample.
TABLE 4 recipe composition for each test group of test two
Note: the protein concentration represents the protein concentration of recombinant human thyrotropin, and the PB buffer represents phosphate buffer.
Subunit oxidation product content detection result and analysis
As shown in FIG. 6, the results of measuring the subunit oxidation product content in thyrotrophin-like products at day 0, day 30 and day 60 at 25. + -. 2 ℃ were obtained using 12 prescriptions of test two. The results show that the subunit oxidation product content is low for formulations with different sodium chloride and mannitol concentrations.
SE-HPLC DETECTION AND ANALYSIS
As shown in FIG. 7, the results of measuring the main peak content in the thyrotrophin-like substance at day 0, day 30 and day 60 at 25. + -. 2 ℃ by SE-HPLC using 12 prescribed samples of the second test described above. As shown in FIG. 8, the results of measuring the amount of the dimer in the thyrotrophin-like substance at day 0, day 30 and day 60 at 25. + -. 2 ℃ by SE-HPLC using 12 prescribed samples of the second test described above. As shown in FIG. 9, the results of measuring the degradation content of thyrotrophin-like substances at day 0, day 30 and day 60 at 25. + -. 2 ℃ by SE-HPLC using 12 prescribed samples of the second test described above. The results show that the formulations of varying sodium chloride and mannitol concentrations have low levels of polymer content and/or degradation, and that the 12 methionine containing formulations of trial two are suitable for storage of the thyrotropin liquid formulations.
Example 3
And (3) test III: effect of different methionine concentrations on the storage Effect of liquid formulations
This example explores the effect of different methionine concentrations on the storage effect of thyrotropin liquid formulations. The prescription composition of each test group for test three is shown in table 5. The liquid formulation was subjected to accelerated stability testing at 25 ± 2 ℃ for 60 days to simulate the effect of long term storage conditions under extreme conditions on the sample.
TABLE 5 recipe composition for each test group of test three
| Prescription | Protein concentration | PB buffer | Mannitol | Sodium chloride | L- | pH | |
| 1 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.1mg/mL | 7.0 | |
| 2 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.2mg/mL | 7.0 | |
| 3 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 1.5mg/mL | 7.0 |
Note: the protein concentration represents the protein concentration of recombinant human thyrotropin, and the PB buffer represents phosphate buffer.
Subunit oxidation product content detection result and analysis
As shown in FIG. 10, the results of measuring the subunit oxidation product content in thyrotrophin-like products at day 0, day 30 and day 60 at 25. + -. 2 ℃ were obtained using the 3 prescriptions of test III. The results show that the concentration of methionine in the range of about 0.1mg/mL to about 1.5mg/mL of the prescribed subunit oxidation product is low.
SE-HPLC DETECTION AND ANALYSIS
As shown in FIG. 11, the results of detecting the main peak content in the thyrotrophin-like substance at day 0, day 30 and day 60 at 25. + -. 2 ℃ by SE-HPLC using the 3 prescribed samples of the third test described above. As shown in FIG. 12, the results of measuring the amount of the dimer in the thyrotrophin-like substance at day 0, day 30 and day 60 at 25. + -. 2 ℃ by SE-HPLC using the 3 prescribed samples of the third test described above. The results show that all formulations containing methionine at concentrations ranging from about 0.1mg/mL to about 1.5mg/mL had low levels of dimer, and that all 3 formulations containing methionine at concentrations ranging from about 0.1mg/mL to about 1.5mg/mL of trial three were suitable for storage of the thyrotropin liquid formulations.
Biological activity assay results and assays
Samples of formula 1 from trial three were tested for relative biological activity of thyrotrophin-like samples at 25 + -2 deg.C on days 0, 30 and 60. On day 0, the relative percent potency of formula 1 for trial three was 102%; on day 30, the relative percent potency of formula 1 for trial three was 102%; on day 60, the relative percent titer of trial three formula 1 was 97%. The results show that the biological activity of formula 1 of the third test is stable at 25 +/-2 ℃, and the liquid preparation of the recombinant human thyrotropin added with methionine can have high physicochemical property and stable biological activity.
Example 4
And (4) testing: effect of different methionine concentrations on the storage Effect of liquid formulations
This example explores the effect of different methionine concentrations on the storage effect of thyrotropin liquid formulations. The prescription composition of each test group for test four is shown in table 6. The liquid formulation was subjected to accelerated stability testing at 40 ± 2 ℃ for 30 days to simulate the effect of long term storage conditions under extreme conditions on the sample.
TABLE 6 prescription composition for each test group of test four
| Prescription | Protein concentration | PB buffer | Mannitol | Sodium chloride | L- | pH | |
| 1 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/ |
0 | 7.0 | |
| 2 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.005mg/mL | 7.0 | |
| 3 | 0.9mg/mL | 20mM | 30mg/ |
2 mg/mL | 0.02mg/mL | 7.0 | |
| 4 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.1mg/mL | 7.0 | |
| 5 | 0.45mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.1mg/mL | 7.0 | |
| 6 | 1.8mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.1mg/mL | 7.0 | |
| 7 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.1mg/mL | 6.5 | |
| 8 | 0.9mg/mL | 20mM | 30mg/mL | 2mg/mL | 0.1mg/mL | 7.5 |
Note: the protein concentration represents the protein concentration of recombinant human thyrotropin, and the PB buffer represents phosphate buffer.
Subunit oxidation product content detection result and analysis
As shown in FIG. 13, the samples of formulas 1, 2, 3 and 4 of test four were used to determine the subunit oxidation product content of thyroxine-like substances at day 0 and day 30 at 40. + -. 2 ℃. The results show that the concentration of methionine in the range of about 0.005mg/mL to about 0.1mg/mL of the prescribed subunit oxidation product is low.
The foregoing detailed description is provided by way of illustration and example, and is not intended to limit the scope of the appended claims. Various modifications of the presently recited embodiments will be apparent to those of ordinary skill in the art and are intended to be within the scope of the appended claims and their equivalents.
Claims (10)
1. A pharmaceutical formulation comprising thyrotropin and methionine, wherein the methionine is at a concentration of about 0.005mg/mL or greater.
2. The pharmaceutical formulation of claim 1, further comprising a buffering agent.
3. The pharmaceutical formulation of claim 1, further comprising a stabilizer.
4. The pharmaceutical formulation of claim 3, the stabilizer being selected from the group consisting of: mannitol and sodium chloride.
5. A kit comprising the pharmaceutical formulation of any one of claims 1-4.
6. A process for the preparation of a pharmaceutical formulation according to any one of claims 1 to 4, comprising the addition of methionine to thyrotropin, the addition of thyrotropin to methionine and/or the simultaneous addition of methionine and thyrotropin.
7. A method of affecting thyroxine levels comprising administering to a subject the pharmaceutical formulation of any one of claims 1-4.
8. A method of affecting the thyroid comprising administering to a subject the pharmaceutical formulation of any one of claims 1-4.
9. A method of affecting thyroid cells comprising administering to a subject the pharmaceutical formulation of any one of claims 1-4.
10. Use of a pharmaceutical formulation of any one of claims 1-4 in the manufacture of a medicament for diagnosing, aiding diagnosis, preventing, aiding treatment, and/or treating a thyroid disorder in a subject.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210212203.1A CN114404575A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
| CN202011417349.7A CN112451652A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011417349.7A CN112451652A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210212203.1A Division CN114404575A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112451652A true CN112451652A (en) | 2021-03-09 |
Family
ID=74801494
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011417349.7A Pending CN112451652A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
| CN202210212203.1A Pending CN114404575A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210212203.1A Pending CN114404575A (en) | 2020-12-07 | 2020-12-07 | Recombinant human thyrotropin injection |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN112451652A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1191139A (en) * | 1997-01-15 | 1998-08-26 | 阿克佐诺贝尔公司 | Liquid gonadotropin containing formulations |
| US20060210557A1 (en) * | 2005-01-28 | 2006-09-21 | Donna Luisi | Stabilized liquid polypeptide formulations |
| US20130171162A1 (en) * | 2008-09-10 | 2013-07-04 | Junyan A. Ji | Compositions and methods for the prevention of oxidative degradation of proteins |
| CN105658201A (en) * | 2013-11-12 | 2016-06-08 | 卡迪拉保健有限公司 | Formulation for gonadotropins |
| US20190062396A1 (en) * | 2015-11-05 | 2019-02-28 | Genexine, Inc. | Composition comprising recombinant human thyroid stimulating hormone and method for producing recombinant human thyroid stimulating hormone |
| US20190060409A1 (en) * | 2016-02-24 | 2019-02-28 | Ferring B.V. | Stable liquid gonadotropin formulation |
| WO2020004368A1 (en) * | 2018-06-25 | 2020-01-02 | Ｊｃｒファーマ株式会社 | Protein-containing aqueous liquid formulation |
-
2020
- 2020-12-07 CN CN202011417349.7A patent/CN112451652A/en active Pending
- 2020-12-07 CN CN202210212203.1A patent/CN114404575A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1191139A (en) * | 1997-01-15 | 1998-08-26 | 阿克佐诺贝尔公司 | Liquid gonadotropin containing formulations |
| US20060210557A1 (en) * | 2005-01-28 | 2006-09-21 | Donna Luisi | Stabilized liquid polypeptide formulations |
| US20130171162A1 (en) * | 2008-09-10 | 2013-07-04 | Junyan A. Ji | Compositions and methods for the prevention of oxidative degradation of proteins |
| CN105658201A (en) * | 2013-11-12 | 2016-06-08 | 卡迪拉保健有限公司 | Formulation for gonadotropins |
| US20190062396A1 (en) * | 2015-11-05 | 2019-02-28 | Genexine, Inc. | Composition comprising recombinant human thyroid stimulating hormone and method for producing recombinant human thyroid stimulating hormone |
| US20190060409A1 (en) * | 2016-02-24 | 2019-02-28 | Ferring B.V. | Stable liquid gonadotropin formulation |
| WO2020004368A1 (en) * | 2018-06-25 | 2020-01-02 | Ｊｃｒファーマ株式会社 | Protein-containing aqueous liquid formulation |
Non-Patent Citations (2)
| Title |
|---|
| 元英进主编: "《现代制药工艺学 上册》", 31 July 2004, 北京：化学工业出版社 * |
| 胡容峰主编: "《工业制剂学》", 31 August 2010, 北京：中国中医药出版社 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114404575A (en) | 2022-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kane et al. | Clozapine: plasma levels and prolactin response | |
| US7148211B2 (en) | Formulation for lipophilic agents | |
| Gilbert et al. | Polyaspartic acid prevents experimental aminoglycoside nephrotoxicity | |
| Roger et al. | Long term treatment of male and female precocious puberty by periodic administration of a long-acting preparation of D-Trp6-luteinizing hormone-releasing hormone microcapsules | |
| FLECHNER et al. | The impact of body weight on cyclosporine pharmacokinetics in renal transplant recipients | |
| Ste‐Marie et al. | Effect of Teriparatide {rhPTH (1‐34)} on BMD When Given to Postmenopausal Women Receiving Hormone Replacement Therapy | |
| EA024152B1 (en) | Topical composition for treating or preventing skin cancer | |
| NO315693B1 (en) | Stable peptide radioisotope composition, as well as process for its preparation, and method for reducing radioactive degradation of the mixture | |
| BRPI0617294A2 (en) | hydroalcoholic gel pharmaceutical composition and use of testosterone | |
| Bearcroft et al. | 5-hydroxytryptamine release into human jejunum by cholera toxin. | |
| Sigalet et al. | Effects of chronic glucagon-like peptide-2 therapy during weaning in neonatal pigs | |
| US20040028736A1 (en) | Absorption enhancers | |
| Jansson et al. | Pharmacokinetic properties and bioavailability of methimazole | |
| Kim et al. | Kinetic analysis of the elimination process of human epidermal growth factor (hEGF) in rats | |
| WO2003006064A1 (en) | Pharmaceutical formulation for the intramuscular administration of fulvestrant | |
| Dromgoole et al. | Tolmetin kinetics and synovial fluid prostaglandin E levels in rheumatoid arthritis | |
| CN112451652A (en) | Recombinant human thyrotropin injection | |
| Gerrits et al. | Oral follicle-stimulating hormone agonist tested in healthy young women of reproductive age failed to demonstrate effect on follicular development but affected thyroid function | |
| Davis et al. | Effects of compounding and storage conditions on stability of pergolide mesylate | |
| Whitford et al. | A comparative study of fluoride pharmacokinetics in five species | |
| Helleberg et al. | Absorption, metabolism and excretion of once-weekly somapacitan, a long-acting growth hormone derivative, after single subcutaneous dosing in human subjects | |
| JP2023549455A (en) | Use of anti-promyostatin/anti-latent myostatin antibodies to treat spinal muscular atrophy | |
| ES2943933T3 (en) | Palonosetron eye drops for treatment or prevention of nausea and vomiting | |
| CN114306218A (en) | R-ketamine pharmaceutical composition for transmucosal administration meeting pharmaceutical antibacterial requirements | |
| Wong et al. | Dose-dependent pharmacokinetics and metabolism of valproic acid in newborn lambs and adult sheep |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210309 |