CN106924721B - Pharmaceutical composition containing human parathyroid hormone for oral mucosal administration - Google Patents
Pharmaceutical composition containing human parathyroid hormone for oral mucosal administration Download PDFInfo
- Publication number
- CN106924721B CN106924721B CN201611190247.XA CN201611190247A CN106924721B CN 106924721 B CN106924721 B CN 106924721B CN 201611190247 A CN201611190247 A CN 201611190247A CN 106924721 B CN106924721 B CN 106924721B
- Authority
- CN
- China
- Prior art keywords
- pharmaceutical composition
- rhpth
- absorption
- mucosa
- parathyroid hormone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 122
- 101001135770 Homo sapiens Parathyroid hormone Proteins 0.000 title claims abstract description 21
- 101001135995 Homo sapiens Probable peptidyl-tRNA hydrolase Proteins 0.000 title claims abstract description 21
- 102000058004 human PTH Human genes 0.000 title claims abstract description 21
- 210000002200 mouth mucosa Anatomy 0.000 claims abstract description 81
- 238000010521 absorption reaction Methods 0.000 claims abstract description 75
- 239000003463 adsorbent Substances 0.000 claims abstract description 41
- 239000003814 drug Substances 0.000 claims abstract description 38
- 239000003961 penetration enhancing agent Substances 0.000 claims abstract description 33
- 210000004877 mucosa Anatomy 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 21
- 208000001132 Osteoporosis Diseases 0.000 claims abstract description 19
- 239000002562 thickening agent Substances 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 9
- 230000008569 process Effects 0.000 claims abstract description 6
- 239000000203 mixture Substances 0.000 claims description 55
- 108090000623 proteins and genes Proteins 0.000 claims description 25
- 102000004169 proteins and genes Human genes 0.000 claims description 24
- 230000002255 enzymatic effect Effects 0.000 claims description 16
- 239000000243 solution Substances 0.000 claims description 13
- 239000011550 stock solution Substances 0.000 claims description 12
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 239000011259 mixed solution Substances 0.000 claims description 6
- 239000007921 spray Substances 0.000 claims description 4
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 3
- 229920001661 Chitosan Polymers 0.000 claims description 3
- 101001041117 Homo sapiens Hyaluronidase PH-20 Proteins 0.000 claims description 3
- 229920002125 Sokalan® Polymers 0.000 claims description 3
- 229960001631 carbomer Drugs 0.000 claims description 3
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 claims description 3
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims description 2
- 230000003232 mucoadhesive effect Effects 0.000 claims 2
- 229920003081 Povidone K 30 Polymers 0.000 claims 1
- 239000001768 carboxy methyl cellulose Substances 0.000 claims 1
- 230000002265 prevention Effects 0.000 claims 1
- 229940120723 recombinant human hyaluronidase Drugs 0.000 claims 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 59
- 229940079593 drug Drugs 0.000 abstract description 28
- 239000003963 antioxidant agent Substances 0.000 abstract description 3
- 230000003078 antioxidant effect Effects 0.000 abstract description 3
- 239000007853 buffer solution Substances 0.000 abstract description 3
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
- 230000003000 nontoxic effect Effects 0.000 abstract description 3
- 239000003381 stabilizer Substances 0.000 abstract description 3
- 239000002085 irritant Substances 0.000 abstract description 2
- 231100000021 irritant Toxicity 0.000 abstract description 2
- 210000002919 epithelial cell Anatomy 0.000 description 32
- 230000003204 osmotic effect Effects 0.000 description 31
- 241000700159 Rattus Species 0.000 description 23
- 230000035515 penetration Effects 0.000 description 22
- 230000001965 increasing effect Effects 0.000 description 20
- 210000000988 bone and bone Anatomy 0.000 description 19
- 102000004190 Enzymes Human genes 0.000 description 18
- 108090000790 Enzymes Proteins 0.000 description 18
- 229940088598 enzyme Drugs 0.000 description 18
- 229920001184 polypeptide Polymers 0.000 description 16
- 108090000765 processed proteins & peptides Proteins 0.000 description 16
- 102000004196 processed proteins & peptides Human genes 0.000 description 16
- 102000004067 Osteocalcin Human genes 0.000 description 13
- 108090000573 Osteocalcin Proteins 0.000 description 13
- 210000002966 serum Anatomy 0.000 description 13
- 108010003272 Hyaluronate lyase Proteins 0.000 description 12
- 102000009066 Hyaluronoglucosaminidase Human genes 0.000 description 12
- 229960002773 hyaluronidase Drugs 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 10
- 230000035699 permeability Effects 0.000 description 10
- 239000000825 pharmaceutical preparation Substances 0.000 description 10
- 210000004400 mucous membrane Anatomy 0.000 description 9
- 239000012981 Hank's balanced salt solution Substances 0.000 description 8
- 230000037182 bone density Effects 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 8
- 238000009472 formulation Methods 0.000 description 8
- 102000003982 Parathyroid hormone Human genes 0.000 description 7
- 108090000445 Parathyroid hormone Proteins 0.000 description 7
- 230000004071 biological effect Effects 0.000 description 7
- 239000000199 parathyroid hormone Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 6
- 239000004480 active ingredient Substances 0.000 description 6
- 230000009286 beneficial effect Effects 0.000 description 6
- 230000037396 body weight Effects 0.000 description 6
- 238000007906 compression Methods 0.000 description 6
- 230000006835 compression Effects 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 238000013001 point bending Methods 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 210000000689 upper leg Anatomy 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 229960001319 parathyroid hormone Drugs 0.000 description 5
- 230000001737 promoting effect Effects 0.000 description 5
- 238000005215 recombination Methods 0.000 description 5
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 4
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 4
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 238000012377 drug delivery Methods 0.000 description 4
- 235000013601 eggs Nutrition 0.000 description 4
- 229920002674 hyaluronan Polymers 0.000 description 4
- 229960003160 hyaluronic acid Drugs 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 230000002195 synergetic effect Effects 0.000 description 4
- 210000000683 abdominal cavity Anatomy 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000004888 barrier function Effects 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 230000017531 blood circulation Effects 0.000 description 3
- 230000037118 bone strength Effects 0.000 description 3
- 229960003964 deoxycholic acid Drugs 0.000 description 3
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 3
- 238000009792 diffusion process Methods 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 210000000214 mouth Anatomy 0.000 description 3
- 230000001009 osteoporotic effect Effects 0.000 description 3
- 239000012466 permeate Substances 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- 229940124531 pharmaceutical excipient Drugs 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 230000006798 recombination Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- OGBMKVWORPGQRR-UMXFMPSGSA-N teriparatide Chemical compound C([C@H](NC(=O)[C@H](CCSC)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)[C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CNC=N1 OGBMKVWORPGQRR-UMXFMPSGSA-N 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 208000010392 Bone Fractures Diseases 0.000 description 2
- 229940124292 CD20 monoclonal antibody Drugs 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 206010015719 Exsanguination Diseases 0.000 description 2
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000008416 bone turnover Effects 0.000 description 2
- 239000006172 buffering agent Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 230000001054 cortical effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 229930004094 glycosylphosphatidylinositol Natural products 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 235000015110 jellies Nutrition 0.000 description 2
- 239000008274 jelly Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000009806 oophorectomy Methods 0.000 description 2
- 210000002997 osteoclast Anatomy 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 229940069328 povidone Drugs 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 229960004641 rituximab Drugs 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000007929 subcutaneous injection Substances 0.000 description 2
- 238000010254 subcutaneous injection Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 210000001694 thigh bone Anatomy 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- -1 AC12F1 Proteins 0.000 description 1
- 102000013563 Acid Phosphatase Human genes 0.000 description 1
- 108010051457 Acid Phosphatase Proteins 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- WQZGKKKJIJFFOK-CBPJZXOFSA-N D-Gulose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O WQZGKKKJIJFFOK-CBPJZXOFSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 101000585728 Homo sapiens Protein O-GlcNAcase Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000015731 Peptide Hormones Human genes 0.000 description 1
- 108010038988 Peptide Hormones Proteins 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 229920000148 Polycarbophil calcium Polymers 0.000 description 1
- 101000713943 Rattus norvegicus Tudor domain-containing protein 7 Proteins 0.000 description 1
- 102000007591 Tartrate-Resistant Acid Phosphatase Human genes 0.000 description 1
- 108010032050 Tartrate-Resistant Acid Phosphatase Proteins 0.000 description 1
- 108010049264 Teriparatide Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 210000004082 barrier epithelial cell Anatomy 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 210000002805 bone matrix Anatomy 0.000 description 1
- 230000018678 bone mineralization Effects 0.000 description 1
- 238000013262 cAMP assay Methods 0.000 description 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 1
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 1
- 210000001715 carotid artery Anatomy 0.000 description 1
- 230000001925 catabolic effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000012669 compression test Methods 0.000 description 1
- 239000013256 coordination polymer Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000004890 epithelial barrier function Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 210000002744 extracellular matrix Anatomy 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
- 230000006870 function Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 102000046319 human OGA Human genes 0.000 description 1
- 229940101556 human hyaluronidase Drugs 0.000 description 1
- 230000009851 immunogenic response Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- PBGKTOXHQIOBKM-FHFVDXKLSA-N insulin (human) Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)[C@@H](C)CC)[C@@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 PBGKTOXHQIOBKM-FHFVDXKLSA-N 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 230000008384 membrane barrier Effects 0.000 description 1
- 230000009245 menopause Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000004898 n-terminal fragment Anatomy 0.000 description 1
- 238000004848 nephelometry Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 210000002655 parathyroid chief cell Anatomy 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229950005134 polycarbophil Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 210000000434 stratum corneum Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 229960005460 teriparatide Drugs 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 239000008215 water for injection Substances 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
- A61K38/29—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
-
- 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
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- 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
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
- A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Endocrinology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Inorganic Chemistry (AREA)
- Nutrition Science (AREA)
- Physiology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to a pharmaceutical composition containing human parathyroid hormone for oral mucosa administration, which comprises high-concentration human parathyroid hormone, an absorption penetration enhancer, a mucosa adsorbent, a thickening agent, a buffer solution, an antioxidant and a stabilizer. The invention also provides a preparation method of the pharmaceutical composition and application of the pharmaceutical composition in preparing medicaments for treating and/or preventing osteoporosis, the pharmaceutical composition is convenient to use, is nontoxic and non-irritant in the using process, takes effect quickly after being used, and can obviously improve the clinical medication compliance of patients.
Description
Technical Field
The invention relates to the field of biomedicine, in particular to a pharmaceutical composition containing human parathyroid hormone for oral mucosa administration, a preparation method thereof and application of the pharmaceutical composition in treating and/or preventing osteoporosis.
Background
Osteoporosis is a common systemic disease characterized by low bone mass, increased bone fragility due to damaged bone microstructure, and easy fracture, and has seriously threatened the health of middle-aged and elderly people, especially postmenopausal women. At present, more than 2 hundred million people all over the world suffer from osteoporosis, and the incidence rate of osteoporosis is the sixth of common diseases and frequently encountered diseases. The osteoporosis patients in China are at the first place in the world, about 9000 ten thousand people have the incidence rate of 15.7 percent in people over 50 years old and 56 percent in people over 60 years old, wherein the incidence rate of postmenopausal women is 60-70 percent higher than that of men, and generally occurs within 5-10 years after menopause of women. With the development of aging of society, the incidence of osteoporosis is on the rise, and is expected to increase to 2.21 billion by 2050.
Parathyroid Hormone (PTH) is a single-chain polypeptide Hormone synthesized and secreted by Parathyroid chief cells, mature PTH contains 84 amino acid residues, and is one of the most important hormones for regulating calcium and phosphorus metabolism and bone turnover in a human body, and small-dose intermittent administration can improve bone density, improve bone quality and reduce the incidence rate of bone fracture. Research shows that the N-terminal fragment PTH (1-34) retains the biological activity of the intact peptide fragment of PTH, can promote bone growth and effectively treat osteoporosis, and the clinical application of PTH (1-34) is mainly used for treating osteoporosis diseases. The current commercial human recombinant PTH (rhPTH (1-34)) injection (Teriparatide) needs daily subcutaneous injection, and frequent injection easily causes great physical psychological and economic burden on patients and influences the administration compliance of the patients. Therefore, changing the administration mode is one of the ways to improve the administration compliance of protein and polypeptide drugs.
The mucosa administration is a common administration mode, and has the advantages of convenient administration, controllable dosage, high patient compliance and the like. The oral mucosa area of the adult is about 200cm2The oral mucosa drug delivery is one of the key research and development directions of protein and polypeptide drug delivery modes.
Due to the special biological characteristics of the oral mucosa, such as the functions of an epithelial barrier, an intraepithelial barrier, a basement membrane barrier and the like, the permeability coefficient of oral mucosa administration is low, general medicines are difficult to enter blood circulation through the oral mucosa, and macromolecular medicines represented by protein and polypeptide medicines are particularly obvious; in addition, mucus secreted from the epithelial surface of the oral cavity and physiological environments such as oral saliva, enzymes, and immune proteins constitute enzymatic and diffusion barriers for protein and polypeptide drugs. Therefore, the low bioavailability of protein and polypeptide drugs is the technical difficulty that needs to be overcome for realizing oral mucosa administration of the drugs. Although oromucosal drug delivery systems have been extensively studied, oromucosal drug delivery systems for proteins and polypeptides have been slow to progress, and there are no protein or polypeptide drugs available on the market that are absorbed through the oromucosa.
The prior art does not disclose oral mucosa administration information of rhPTH (1-34), and the rhPTH (1-34) is known to be a biological macromolecular polypeptide drug, is greatly influenced by the oral mucosa structure and the oral physiological environment in the oral mucosa administration process, is not easy to be directly absorbed into blood circulation through the oral mucosa, and has great technical difficulty in oral mucosa administration. Thus, it is an unsolved technical problem of the prior art to obtain a formulation of rhPTH (1-34) which is administered through the oral mucosa and can obtain the desired therapeutic effect.
Disclosure of Invention
The invention aims to solve the defects of the prior art and initially creates a pharmaceutical composition containing human parathyroid hormone rhPTH (1-34) for oral mucosa administration, wherein the active ingredients of the pharmaceutical composition can permeate oral mucosa, and the pharmaceutical composition has the characteristic of high penetration and absorption, is beneficial to improving the bioavailability of the medicament and can realize the oral mucosa administration of the human parathyroid hormone rhPTH (1-34).
The purpose of the invention is realized by the following technical scheme:
a pharmaceutical composition containing human parathyroid hormone for oral mucosa administration is a solution system, and comprises human parathyroid hormone rhPTH (1-34) and absorption penetration enhancer PH 20.
Specifically, the human parathyroid hormone is recombinant human parathyroid hormone (rhPTH (1-34)), which can be prepared by a known genetic engineering recombination technique. The mass of the rhPTH in the pharmaceutical composition is 5-2000 mug, the rhPTH can be present in the pharmaceutical composition at a suitable concentration, preferably, the concentration of the rhPTH in the pharmaceutical composition is 50-2000 mug/ml, and more preferably, the concentration of the rhPTH in the pharmaceutical composition is 100-1000 mug/ml.
The inventors have surprisingly found during the course of experiments that there is a synergistic effect of recombinant human parathyroid hormone (rhPTH (1-34)) and an absorption penetration enhancer. Specifically, when a specific type of absorption penetration enhancer with enzymatic activity is adopted, the oral mucosa penetration effect of the effective component can be obviously improved, the improvement of the bioavailability of the medicament is facilitated, and the oral mucosa administration is further realized to have the characteristics of quick response and the like.
The absorption penetration enhancer is hyaluronidase, the hyaluronidase is PH20, preferably human hyaluronidase PH20(hPH20), and the hyaluronidase can be prepared by the prior known genetic engineering recombination technology.
Because the respective properties of the protein/polypeptide drugs are different, and the influence degrees of the protein/polypeptide drugs on the oral mucosa structure and the oral physiological environment are different, not all the protein/polypeptide drugs are suitable for oral mucosa administration, and not all the protein/polypeptide drugs for oral mucosa administration can achieve the expected treatment effect. Therefore, the technical key to achieve the above objects of the present invention is the type of absorption penetration enhancer and the contents of rhPTH (1-34) and absorption penetration enhancer.
Specifically, rhPTH (1-34) belongs to hydrophilic polypeptide, has the molecular weight of 4.9KD, belongs to macromolecular drugs, is greatly influenced by the structure of oral mucosa and the physiological environment of the oral cavity, and is difficult to permeate into blood to be absorbed through epithelial cells of the oral mucosa when being singly used. In order to allow rhPTH (1-34) to pass through the oral mucosa and enter the capillaries, the amount of penetration is increased to achieve a therapeutically effective concentration of the drug, and an appropriate absorption enhancer is added to the pharmaceutical composition. The types of absorption penetration promoters are selected, the prior art discloses a plurality of compounds with penetration promoting effects, such as hyaluronidase (PH20), cholate, fatty acid, alcohol, surfactant and the like, factors such as penetration promoting effect, compatibility, adverse reaction and the like need to be comprehensively considered for the selection of the absorption penetration promoters, partial penetration promoters have poor penetration promoting effect and cannot achieve the purpose of treatment, the partial penetration promoters cause irreversible damage or even degradation to oral mucosa after long-term use, so that adverse reactions such as inflammation and the like can occur, each absorption penetration promoter has good and bad effects, and the hyaluronidase (PH20) has the best penetration promoting effect on rhPTH (1-34).
Specifically, the hyaluronidase (PH20) belongs to mucopolysaccharide catabolic enzyme, which is generally neutral or acidic, can hydrolyze hyaluronic acid in intercellular matrix, and can form extracellular matrix pore canals less than 200nm in a short time, so that the viscosity of oral mucosa is reduced, the permeability is increased, the rapid diffusion and permeation of rhPTH (1-34) in the mucosa are promoted, the absorption effect of capillary vessels is enhanced, and the bioavailability of rhPTH (1-34) is improved. Meanwhile, PH20 can be rapidly cleared by plasma, the half-life period of the plasma is short, and the hyaluronic acid in the mucosa interstitium can be completely rebuilt within 24-48h, and the integrity of the mucosa structure can be rapidly recovered. Therefore, hyaluronidase (PH20) reversibly alters the stratum corneum barrier function of oral mucosa to promote mucosal osmotic absorption of rhPTH (1-34), and damaged mucosa can recover more rapidly without causing serious irritation and damage to oral mucosa. The hyaluronidase (PH20) of the present invention is preferably human hyaluronidase (hPH 20). Furthermore, the invention adopts soluble hPH20(rhPH20) prepared by gene recombination technology as an absorption penetration enhancer, in particular, rhPH20 is mature PH20 polypeptide, the C-end of the polypeptide lacks all or part of Glycosyl Phosphatidylinositol (GPI) attachment sites, so that the polypeptide is not combined with CHO cell membrane after being expressed in Chinese Hamster Ovary (CHO) cells in a recombination way, thereby leading rhPH20 to be secreted and dissolved in a cell culture medium.
The amount of the absorption penetration enhancer, hyaluronidase (PH20), is controlled within a reasonable range. Specifically, the enzymatic activity of the absorption penetration enhancer in the pharmaceutical composition is 500-20000U, and the absorption penetration enhancer can be present in the pharmaceutical composition at an appropriate enzymatic activity concentration, preferably, the enzymatic activity concentration of the absorption penetration enhancer is 500-20000U/ml, and more preferably, the enzymatic activity of the absorption penetration enhancer is 1000-20000U/ml.
The inventor further discovers in the experimental process that the recombinant human parathyroid hormone (rhPTH (1-34)), the absorption penetration enhancer and the mucosa adsorbent have the synergistic effect, and the optimization of the oral mucosa administration effect is favorably realized.
The mucosa adsorbent in the pharmaceutical composition refers to a pharmaceutical excipient which has a viscosity regulating effect to ensure that the pharmaceutical composition is in contact with oral mucosa. The mucous membrane/saliva is secreted by the oral mucosa, so that the pharmaceutical composition is diluted and partially swallowed and cannot achieve the treatment effect, and after the mucous membrane adsorbent is added, the mucous membrane adsorbent can absorb water in the mucous membrane when contacting the oral mucosa, and contacts and adheres to the oral mucosa through physical action, so that the composition is ensured to stay in the oral cavity and can fully contact the oral mucosa, and the influence of the oral environment on the active ingredients of the pharmaceutical composition is reduced, therefore, the mucous membrane adsorbent is added into the pharmaceutical composition to ensure the stability of the pharmaceutical composition in the oral environment, the utilization rate of rhPH20 is further improved, and the permeation of rhPTH (1-34) through the oral mucosa is further improved. Generally, the aforementioned pharmaceutical excipients having viscosity regulating effect to ensure the pharmaceutical composition to contact with the oral mucosa have the synergistic effect, and preferably, the mucosa adsorbent may be one or more of povidone, Carbomer (CP), carboxymethyl chitosan, polyvinyl acid, and polycarbophil mixed in any ratio. Those skilled in the art can use a proper amount of mucosa adsorbent to achieve the above purpose of further improving the permeation of rhPTH (1-34) in the pharmaceutical composition through mucosa, wherein the mass ratio of the mucosa adsorbent in the pharmaceutical composition is in the range of 0.1-99.9%, the mucosa adsorbent can be present in the pharmaceutical composition at a proper concentration, and preferably, the concentration of the mucosa adsorbent in the pharmaceutical composition is 0.2-15 mg/ml.
It can be seen that the invention can realize the purpose of remarkably improving the permeation of rhPTH (1-34) to oral mucosa in oral mucosa administration by selecting the type of penetration enhancer and further selecting the type of mucosa adsorbent, thereby further realizing the characteristics of quick effect taking and good treatment effect of oral mucosa administration of human parathyroid hormone, good patient compliance and the like. One or more than two auxiliary materials such as a thickening agent, a buffering agent, an antioxidant, a stabilizing agent and the like can be further added into the pharmaceutical composition according to specific needs so as to further solve the technical problems of viscosity, stability and the like of the pharmaceutical composition and further optimize various performances such as the drug forming property and the like of the pharmaceutical composition. For example, the activity and purity of the enzyme in the resulting pharmaceutical composition may be further improved by adding a suitable amount of thickener carboxymethylcellulose sodium (CMC-Na).
Another object of the present invention is to provide a method for preparing the above pharmaceutical composition containing rhPTH (1-34) for oromucosal administration, which comprises the steps of:
1) preparing a mixed solution containing components other than rhPTH (1-34) and an absorption penetration enhancer;
2) adding the protein stock solution of the absorption penetration enhancer obtained by recombinant preparation into the mixed solution obtained in the step 1), and uniformly mixing;
3) adding rhPTH (1-34) protein stock solution obtained by recombinant preparation into the mixed solution obtained in the step 2), and uniformly mixing;
4) filtering and sterilizing to obtain the pharmaceutical composition containing rPTH (1-34) for oral mucosal administration.
Wherein, the mixed solution in the step 1) is an aqueous solution, and the components are consistent with those in the previous invention, and may contain or not contain a mucous membrane adsorbent, and may further contain one or more than two pharmaceutical excipients selected from a viscous agent, a buffering agent, an antioxidant, a stabilizer and the like on the basis.
The method is beneficial to the industrialized mass production of the pharmaceutical composition containing the rhPTH (1-34) and ensures the stability of the properties of the effective components and other components/auxiliary materials in the production process.
The third purpose of the invention is to provide a pharmaceutical preparation containing the rPTH (1-34) pharmaceutical composition, which is prepared from the rPTH (1-34) pharmaceutical composition. The pharmaceutical preparation comprises, but is not limited to, a spray, a sticking film agent and the like.
The pharmaceutical preparation containing the rPTH (1-34) pharmaceutical composition is prepared from the rPTH (1-34) pharmaceutical composition and is a pharmaceutical preparation for clinical use. The pharmaceutical preparation can be used for oral mucosa administration, has the characteristics of quick administration and effect taking, convenient administration and the like, and has good patient compliance.
The fourth purpose of the invention is to provide the application of a pharmaceutical preparation containing the rhPTH (1-34) pharmaceutical composition for oral mucosal administration in preparing the drugs for treating and/or preventing osteoporosis.
The pharmaceutical composition contains the absorption penetration enhancer and the mucosa adsorbent with specific types and dosage ranges, so that the pharmaceutical active substance rhPTH (1-34) can quickly permeate and absorb through oral mucosa to enter blood circulation, effective blood concentration is achieved, administration times can be reduced, administration pain is avoided, and clinical medication compliance of patients is remarkably improved.
Compared with the prior art, the invention has the following advantages and beneficial effects:
1. the pharmaceutical composition contains rhPTH (1-34) and can be used for oral mucosa administration, the pharmaceutical composition improves the permeability of rhPTH (1-34) to oral mucosa by adding specific types and specific amounts of absorption penetration enhancer and mucosa adsorbent, is beneficial to improving the bioavailability of the medicament, further realizes the characteristics of quick oral mucosa administration effect, good patient compliance and the like of human parathyroid hormone, and achieves the expected treatment effect;
2. provides a preparation method of a pharmaceutical composition containing rhPTH (1-34) for oral mucosa administration, which is suitable for industrial mass production and ensures the stability of the properties of active ingredients and other ingredients/auxiliary materials in the production process;
3. the invention provides a pharmaceutical preparation containing the rPTH (1-34) pharmaceutical composition, the pharmaceutical preparation is prepared from the rPTH (1-34) pharmaceutical composition and can be used for oral mucosa administration, and the pharmaceutical preparation has the characteristics of quick response, convenient administration and the like of the traditional oral mucosa administration and good patient compliance;
4. the application of the medicinal composition containing rhPTH (1-34) for oral mucosa administration in preparing the medicament for treating and/or preventing osteoporosis is provided, the formula of the medicinal composition is safe and non-toxic, the use is convenient, the medicinal composition is non-toxic and non-irritant in the use process, the medicament effect is quick, and the clinical medication compliance of patients is obviously improved.
Drawings
FIG. 1 is a graph of the effect of different enzymatic activities of the absorption penetration enhancer rhPH20 on the osmotic absorption of rhPTH (1-34) by TR146 oral mucosal epithelial cells (●, ■ and ▲ indicate different concentrations of rhPTH (1-34), 100, 560 and 1000. mu.g/ml, respectively);
FIG. 2 is a graph of the effect of different concentrations of adhesive on the osmotic absorption of rhPTH (1-34) by TR146 oral mucosal epithelial cells (●, ■, and ▲ represent different concentrations of rhPTH (1-34), 100, 560, and 1000 μ g/ml, respectively).
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the invention are not limited thereto.
Examples the protein stock solutions containing rhPTH (1-34) described in the literature: sung wl.j biolchem.1991; 266(5) 2831-5, the obtained protein stock solution contains rhPTH (1-34) with the concentration of 2mg/ml and the purity of 98.0 percent;
examples the protein stocks containing rhPH20 were obtained from the literature: CN200680013224.X the protein stock solution obtained by the method disclosed in examples 2-9 contained rhPH20 at a concentration of 2mg/ml and a purity of 98.0%.
Examples said CD20 was prepared as disclosed in example II of document US5736137, and the resulting protein stock solution contained rhPH20 at a concentration of 2mg/ml and a purity of 98.0%.
Example 1
Pharmaceutical composition containing rhPTH (1-34)
TABLE 1 pharmaceutical composition A-X with various component ratios
The rhPTH (1-34) protein stock solution (with the purity of 98 percent and the concentration of 2mg/ml) and the rhPH20 protein stock solution (with the purity of 98 percent and the concentration of 2mg/ml) which are prepared by recombination are respectively unfrozen at room temperature or 2-8 ℃, and are stored at 2-8 ℃ after unfreezing.
Accurately weighing mucosa adsorbent according to the ratio of each component of the pharmaceutical composition A-X in Table 1, dissolving with water for injection to obtain a mixed solution, adding unfrozen rhPH20 protein stock solution, mixing, adding rhPTH (1-34) protein stock solution, stirring, mixing, filtering with a filter membrane with pore diameter of 0.22 μm under aseptic condition, and sterilizing to obtain the pharmaceutical composition A-X.
According to the production requirement of the preparation, the obtained medicine composition can be further subpackaged into medicine units containing 50-2000 mu g of rhPTH (1-34) according to the requirement.
Comparative example 1, pharmaceutical composition containing rhPTH (1-34) and Process for preparing the same
The components in the following formula A1-A5 shown in the following Table 2 are mixed, wherein the composition A1 does not contain an absorption penetration enhancer rhPH20, a thickening agent CMC-Na and a mucous membrane adsorbent, and the other components and the mixture ratio are the same as those of the composition A; the composition A2 contains the same components and proportions as those of the composition H except that the composition A2 does not contain an absorption penetration enhancer rhPH 20; the composition A3 contains the same components and proportions as those of the composition Q except that the thickening agent CMC-Na and the mucous membrane adsorbent are not contained; the composition A4 contains the same components and proportions as those of the composition T except that the composition A4 does not contain an absorption penetration enhancer rhPH 20; composition A5A 1-A5 was prepared in the same manner as in example 1, except that the thickener CMC-Na and the mucous membrane adsorbent were not included, and the other components and their proportions were the same as those of composition X.
TABLE 2 formulation of pharmaceutical composition A1-A5
Comparative example 2
Pharmaceutical composition containing CD20
The pharmaceutical composition containing CD20 was prepared from CD20 as the active ingredient according to the formulation and method of composition B1 in table 3 below using the same formulation and method as composition V in example 1.
TABLE 3 pharmaceutical compositions containing anti-CD 20 monoclonal antibodies
Comparative example 3
Sodium deoxycholate is used as an absorption-promoting penetrant, and the pharmaceutical composition C1-C4 containing rhPTH (1-34) is prepared according to the mixture ratio of the components in the following table 4 by the same method as that in the example 1.
TABLE 4 formulation of pharmaceutical composition C1-C4
Example 2 osmotic absorbency test
The osmotic absorbability of the pharmaceutical compositions obtained in example 1 and comparative examples 1-4 was examined according to a literature report (Porter A. pharm Res,2002,19(2): 169-.
DMEM culture solution in each chamber above and below the plug-in culture dish is sucked off, and TR146 cells are washed by HBSS buffer solution. Then adding HBSS solution into upper and lower chambers respectively, after balancing for 1h, sucking out HBSS solution in upper chamber, and adding 2mL test sample (pharmaceutical composition A-X, pharmaceutical composition C1-C4). 100 μ L of sample was taken from the lower chamber at the set time and the same amount of HBSS solution was immediately replenished. And (3) adopting a chromatographic method, carrying out sample injection analysis, recording the obtained peak area (A), and calculating the accumulated permeation quantity of the rhPTH (1-34) at each time point. The following indices were calculated from the data obtained: (1) apparent permeability coefficient of rhPTH (1-34): papp＝(dQ/dt)/(A×C0) (ii) a (2) Permeability coefficient of rhPTH (1-34) through the plug-in culture dish (cell-free): pPTH＝(dQ/dt)/(A×C0) (ii) a (3) Cell permeability coefficient: 1/Papp＝1/Pc+1/PPTH(ii) a (4) Osmotic absorption enhancement ratio: ER ═ Pc (composition)/Pc (control). Where dQ/dt represents the steady state permeation rate and A is the permeation area (cm)2)，C0Refers to the initial concentration of upper chamber rhPTH (1-34), PcRefers to the cell-penetrating system of rhPTH (1-34)Number, PPTHMeaning the osmotic coefficient of the inserted culture dish in the absence of cells, Pc (composition)Refers to the cell permeability coefficient of rhPTH (1-34) in the pharmaceutical composition A-X through TR146, Pc (control)Refer to the cell permeability coefficient of rhPTH (1-34) through TR146 in composition A1-A5, where controls for pharmaceutical composition A, H, Q, T, X were composition A1, A2, A3, A4, and A5, respectively.
When rhPH20 and rhPTH (1-34) are used respectively, the method for detecting the permeability of the oral mucosa epithelial cells of the rhPTH (1-34) through TR146 is as follows: DMEM culture solution in each chamber above and below the plug-in culture dish is sucked off, and TR146 cells are washed by HBSS buffer solution. Then HBSS solution was added to the upper and lower chambers, respectively, after equilibration for 1 hour, the HBSS solution in the upper chamber was aspirated, and 1mL of rhPH20 having an enzymatic activity of 1000U/mL was added, followed by addition of rhPTH (1-34) having a concentration of 100. mu.g/mL). 100 μ L of sample was taken from the lower chamber at the set time and the same amount of HBSS solution was immediately replenished. And (3) adopting a chromatographic method, carrying out sample injection analysis, recording the obtained peak area (A), and calculating the accumulated permeation quantity of the rhPTH (1-34) at each time point.
(1) Effect of each of the pharmaceutical compositions of example 1 and comparative example 1 on the osmotic absorbability of rhPTH (1-34) through TR146 oral mucosal epithelial cells:
table 5. effect of each of the pharmaceutical compositions of example 1 and comparative example 1 on the osmotic absorbability of rhPTH (1-34) through TR146 oral mucosal epithelial cells (x ± s, n ═ 3)
As shown in Table 5, when the activity of the absorption penetration enhancer rhPH20 enzyme is 1000-20000U/ml and the concentration of the mucosa adsorbent is 0.3-15 mg/ml, rhPTH (1-34) in the pharmaceutical composition A-X has good penetration absorption effect through the oral mucosa epithelial cells of TR 146; composition a1 lacks the absorption penetration enhancer rhPH20 and mucosal adsorbents, such that the osmotic absorption of rhPTH (1-34) is significantly lower than that of pharmaceutical composition a; compositions a2, a4 contained mucosal adsorbents such that the osmotic absorption of rhPTH (1-34) by TR146 oral mucosal epithelial cells was slightly higher than composition a1, but due to the lack of absorption penetration enhancer rhPTH20, the osmotic absorption of rhPTH (1-34) was significantly lower than composition H, T; the compositions A3 and A5 contain an absorption penetration enhancer rhPH20, can degrade the connection of intercellular substance, and enlarge the aperture of the cells, thereby improving the penetration absorption effect of rhPTH (1-34) through TR146 oral mucosa epithelial cells, and are higher than the compositions A1, A2 and A4, but are still significantly lower than the composition Q, X. Therefore, the absorption penetration enhancer rhPH20, the mucosa adsorbent and the thickening agent have synergistic effect on the penetration absorption of rhPTH (1-34) by the oral mucosa epithelial cells of TR 146.
When rhPH20 (enzyme activity 1000U/ml) and rhPTH (1-34) (concentration 100 μ g/ml) were placed in the upper chamber, respectively, the penetration and absorption of rhPTH (1-34) through TR146 oral mucosa epithelial cells (Pc value of (2.37. + -. 0.31). times.10-7m/s) is higher than composition A1, but is still significantly lower than composition A (the ER value is 7.8 +/-1.26 relative to composition A), which indicates that rhPH20 and the effective component rhPTH (1-34) respectively can promote the osmotic absorption of the rhPTH (1-34) through TR146 oral mucosal epithelial cells to a certain extent, but is still significantly lower than the osmotic absorption of the pharmaceutical composition of the invention to the rhPTH (1-34);
(2) for comparative example 2, the active ingredient is anti-CD 20 mab, such as rituximab, and to achieve oral mucosal administration, the composition can be prepared according to the ratio of the ingredients in composition B1 in table 2 by the same method as in example 1. However, in order to prepare a composition containing rituximab at such a high concentration, concentration of the antibody is required, but the concentrated antibody is liable to aggregate, the antibody activity cannot be ensured, and after absorption through the oral mucosa, the immunogenic reaction of the therapeutic antibody is liable to occur in vivo, and the neutralizing antibody produced in this process may render the therapeutic antibody ineffective. Therefore, the preparation of the pharmaceutical composition containing the anti-CD 20 monoclonal antibody for oral mucosal administration and the use thereof in treating related diseases cannot be realized technically.
(3) In comparative example 3, when sodium deoxycholate was used as an absorption permeation enhancer of the pharmaceutical composition, the ordered structure of lipid bilayers of TR146 oral mucosal epithelial cells was changed to decrease the order of phospholipid molecules, thereby decreasing the viscosity between TR146 oral mucosal epithelial cells and increasing the permeability between the mucosal cells, but the pore size formed between TR146 oral mucosal epithelial cells was limited, and was not as large as that formed after rhPH20 degraded the hyaluronic acid between mucosal cells, and the effective concentration of rhPTH (1-34) could not be reached even when the amount of sodium deoxycholate was increased. Therefore, the penetration and absorption of the rhPTH (1-34) through TR146 oral mucosa epithelial cells are lower than that of the rhPTH (1-34) in the example 1, and the rhPTH (1-34) can not be administered through the oral mucosa to achieve the effective concentration of the medicament.
Example 3 osmotic Effect of different enzymatic Activity of absorption penetration enhancers rhPH20 on the osmotic absorption of rhPTH (1-34) by TR146 oral mucosal epithelial cells
According to the following composition proportioning of D1-D10 in the following table 6.1, wherein the enzymatic activities of absorption penetration enhancers rhPH20 are respectively 300, 600, 1000, 2000, 5000, 8000, 12000 and 20000U/ml, the same method as that of example 1 is adopted to prepare the composition containing rhPH20 with different enzymatic activities, and the same method as that of example 2 is used to detect the influence of rhPH20 with different enzymatic activities on the penetration and absorption of rhPTH (1-34) through TR146 oral mucosa epithelial cells.
The experimental results are shown in Table 6.2 and figure 1, and the pharmaceutical composition D1-D10 can effectively promote the permeation and absorption of rhPTH (1-34) through TR146 oral mucosal epithelial cells when the activity of the rhPH20 enzyme is 1000-20000U/ml; when the activity of the rhPH20 enzyme is less than 1000U/ml, the osmotic absorption effect of the rhPTH (1-34) through the TR146 oral mucosa epithelial cells is lower than that of the rhPH20 enzyme when the activity of the rhPH20 enzyme is 1000-20000U/ml, and particularly when the activity of the rhPH20 enzyme is less than 500U/ml, the osmotic absorption of the rhPTH (1-34) through the TR146 oral mucosa epithelial cells is very low, so that the bioavailability of the rhPTH (1-34) cannot be obviously increased; when the activity of rhPH20 enzyme is higher than 20000U/ml, the osmotic absorption effect of rhPTH (1-34) is not increased significantly any more. Therefore, when the enzyme activity of the rhPH20 is 1000-20000U/ml, the osmotic absorption of the rhPTH (1-34) through TR146 oral mucosa epithelial cells has dose dependence.
As shown in figure 1, when the concentration of the effective component rhPTH (1-34) of the pharmaceutical composition is 100 and 1000. mu.g/ml, the species and the proportion of the mucosa adsorbent are the same as those of the pharmaceutical composition D1-D10 shown in Table 6.1, and the activity of rhPH20 enzyme is 300-30000U/ml, the osmotic absorption curve of the oral mucosa epithelial cells of rhPTH (1-34) through TR146 is similar to that of the pharmaceutical composition D1-D10.
TABLE 6.1 formulation of pharmaceutical composition D1-D10
TABLE 6.2 osmotic absorbability of pharmaceutical composition D1-D10rhPTH (1-34) through TR146 oral mucosal epithelial cells
Example 4 osmotic absorption Effect of different concentrations of mucosal adsorbents on oral mucosal epithelial cells of rhPTH (1-34) Via TR146
According to the following formulation of compositions E1-E11 in Table 7.1, wherein the mucosal adsorbent comprises povidone, carbomer and carboxymethyl chitosan, and the concentrations thereof are 0.05, 0.1, 0.2, 0.8, 1, 5, 7, 10, 15, 18 and 25mg/ml, the same method as that of example 1 is adopted to prepare compositions containing mucosal adsorbents with different concentrations, and the influence of the mucosal adsorbents with different concentrations on the osmotic absorption of rhPTH (1-34) through TR146 oral mucosal epithelial cells is tested by the same method as that of example 3.
The experimental results are shown in table 7.2 and fig. 2, when the concentration of the mucosa adsorbent is 0.2-15mg/ml, the pharmaceutical composition E1-E11 can effectively promote the composition to adhere to the oral mucosa, is beneficial to the osmotic absorption of rhPTH (1-34) through the epithelial cells of the oral mucosa of TR146, and can improve the bioavailability; when the concentration of the mucosa adsorbent is less than 0.2mg/ml, the osmotic absorption effect of the rhPTH (1-34) on the oral mucosa epithelial cells of TR146 is very low, which indicates that the concentration of the mucosa adsorbent is too low to be beneficial to the adhesion effect of the composition, so that the osmotic absorption effect of the rhPTH (1-34) is obviously reduced; when the concentration of the mucosa adsorbent is more than 15mg/ml, the osmotic absorbability of the rhPTH (1-34) is not remarkably increased, which shows that when the concentration of the mucosa adsorbent is more than 15mg/ml, the viscosity of the composition is remarkably increased, so that the adhesion is high when the composition is contacted with oral mucosa, and the diffusion and absorption effects of the rhPTH (1-34) on the epithelial cells of the oral mucosa of TR146 are influenced. Therefore, the concentration of the mucosa adsorbent is 0.2-15mg/ml, and the osmotic absorption of the rhPTH (1-34) through TR146 oral mucosa epithelial cells has dose dependence.
TABLE 7.1 formulation of pharmaceutical composition E1-E11
TABLE 7.2 osmotic absorbability of pharmaceutical composition E1-E11rhPTH (1-34) through TR146 oral mucosal epithelial cells
As shown in figure 2, when the concentration of the effective component rhPTH (1-34) of the pharmaceutical composition is 100 and 1000. mu.g/ml respectively, the activity of rhPH20 enzyme is 1000 and 12000U/ml respectively, and the concentration of the mucosa adsorbent is 0.05-25 mg/ml, the osmotic absorption curve of the oral mucosa epithelial cells of the rhPTH (1-34) through TR146 is similar to that of the pharmaceutical composition E1-E11.
Example 4 biological activity:
(1) biological Activity of rhPTH (1-34)
The biological activity of rhPTH (1-34) in the pharmaceutical composition obtained in example 1 was measured by cAMP assay, and the purity and concentration of rhPTH (1-34) were measured by RP-HPLC, as shown in Table 8:
TABLE 8 biological Activity, purity and concentration of rhPTH (1-34) in pharmaceutical compositions A-X of example 1
It is known that when the activity of the absorption-promoting penetrant rhPH20 is 1000-2000 IU/ml, the concentration of the mucosa adsorbent is 0.2-15mg/ml, and/or the concentration of the thickener is 0.3-4 mg/ml, the purity of rhPTH (1-34) in the pharmaceutical composition obtained in example 1 is above 98%, the biological activity is kept at a high level, and the concentration is basically maintained. In addition, the biological activity and purity of rhPTH (1-34) can be further improved by adding the thickening agent.
(2) Activity test of absorption-promoting penetrant (rhPH 20):
the enzymatic activity of rhPH20 in the above-mentioned pharmaceutical compositions a-X was examined according to the method for measuring the activity of hyaluronidase (nephelometry) provided in appendix XII C of the second part of the chinese pharmacopoeia, 2010 edition.
The determination method is based on the fact that hyaluronic acid reacts with serum to form stable jelly under acidic conditions, the jelly formed by reaction is reduced and becomes clear after being degraded by hyaluronidase, the clarity is measured by the wavelength of 640nm, and the content of hyaluronidase and the clarity are in direct proportion.
The enzyme activity of rhPH20 was calculated from a standard curve generated by measuring dilutions of the reference standard with rhPH20, and the results are shown in table 9:
TABLE 9 enzymatic Activity of rhPH20 in pharmaceutical compositions A-X
As can be seen from the above data, the rhPH20 enzyme activity in the pharmaceutical composition obtained in example 1 remained high and was approximately 98% pure; the enzymatic activity and purity of rhPH20 were further improved by the addition of a thickening agent.
EXAMPLE 5 in vivo efficacy test of pharmaceutical composition containing rhPTH (1-34) administered through oral mucosa
SPF grade SD female rats 90, 6 months old, 257 ± 13 grams body weight. Of these, 10 were baseline, 20 were used for model identification, and 60 were used for pharmacodynamic studies.
Experimental animals were either fully bilateral ovariectomized (ovariectomized group, Ovx) or dissected 20 animals with only a small amount of adipose tissue surrounding the ovaries (Sham-extirpated egg group, Sham) by abdominal injection of 60 animals anesthetized lower dorsal incisions with ketamine (100mg/kg body weight) prior to surgery as a baseline group (Base).
Animals in the Base group (Base) were anesthetized with 22% ulinase (0.5ml/100g body weight) in the abdominal cavity and sacrificed by carotid exsanguination. Taking thighbone and lumbar vertebra to measure bone mass and bone biomechanical property.
After 12 weeks of surgery, 10 eggs were randomly picked from the Sham egg group (Sham) and the ovariectomy group (Ovx), anesthetized with 22% urothane abdominal cavity, and sacrificed by carotid exsanguination. Weighing uterus, and measuring bone mass and bone biomechanical property of thighbone and lumbar vertebra to confirm that the osteoporosis model of the ovariectomized rat is established.
The remaining experimental animals were further divided into groups of 10 animals each. The remaining rats of the Sham egg group (Sham) served as normal control group (Sham), and the remaining rats of the ovariectomy group (Ovx) were further divided into 5 groups in parallel, including negative control group (NS), drug to be tested group (ORAL) which was the pharmaceutical composition W of example 1 at different dosages, and positive control group (SC) for subcutaneous injection.
Normal control (SHAM) and negative control (NS) rats were given vehicle via oral mucosa as control treatment; three test drug groups (ORAL) were administered 463, 1390 and 4170. mu.g/kg of pharmaceutical composition W (corresponding to rhPTH (1-34) of 7, 21 and 63. mu.g/kg., respectively, referring to the previously studied insulin spray Oral-lyn set dose-fold relative to the injection administration); the positive control group (SC) was administered rhPTH (1-34) 2.1. mu.g/kg/time. Each group was treated 6 times per week for 17 weeks. The experimental groups and treatment details are shown in table 10 below.
After rats in a normal control group (SHAM), a negative control group (NS) and three groups of drug groups W (ORAL) to be tested with different administration doses are anesthetized by injecting 60mg/kg sodium pentobarbital solution into tail veins, the rats are kept in a supine position on an anesthesia table and are ligated by an esophagus, and each experimental animal is ensured to be at the same position on a horizontal line during oral mucosa administration. Different drugs of a normal control group (SHAM), a negative control group (NS) and three groups of drug groups to be tested (ORAL) are loaded in different spray bottles, and then ORAL mucosa administration is carried out on the animals correspondingly grouped.
TABLE 10 Experimental groupings and contents
Detection indexes are as follows:
injecting tetracycline (30mg/Kg of body weight) and calcein (10mg/Kg of body weight) into abdominal cavity 14 days and 4 days before the treatment to prepare bone tissue morphology measurement specimen, and analyzing mineralization deposition rate. After the treatment, rats were anesthetized by intraperitoneal injection with 22% of gulose (0.5ml/100g of body weight), blood was collected from the carotid artery, and then the femoral bone, the tibia, the lumbar and other bone samples and other samples were taken after sacrifice for the following measurement.
(1) Bone density (BMD): the femoral and lumbar vertebral density values reflect the bone density of predominantly cortical and cancellous bone, respectively. Femoral (left) and lumbar (L1-5) bone density was scanned at high resolution using a dual energy X-ray bone densitometer (local Discovery A, accuracy 1%; CV < 1%) to obtain its bone density value (g/cm 2).
(2) Bone biomechanics: the femur three-point bending test and the lumbar compression test respectively mainly reflect the mechanical properties of cortical bone and cancellous bone. After muscle is removed from fresh femur (right) and lumbar vertebra (L3) specimens, the specimens are respectively tested on a three-point bending device (span 18 mm; speed 10.0 mm/min; temperature 23 ℃, humidity 60-70%) and a compression device of a universal material tester (INSTRON-5543 USA; precision 0.04%; measurement range 10-1100N), and three-point bending maximum load and compression maximum load are obtained, and the result is expressed in Newton force (N).
(3) Biochemical indexes of bone conversion: including serum OCN, and TRAP5 b. Osteocalcin (OCN) is secreted primarily by mature osteoblasts, deposited in the bone matrix, and serum levels reflecting bone turnover levels and the degree of bone mineralization, and tested using the Rat OCN enzyme immunoassay kit (Rat-Mid Osteocalcin, AC12F1, IDS, UK). Tartrate-resistant acid phosphatase 5b (TRAP5b) is produced mainly by osteoclast secretion, and its serum activity reflects osteoclast number and activity, and TRAP5b activity was measured in rat serum using rat TRAP5b enzyme immunoassay kit (rattrap assay, SB-TR102, IDS).
The experimental results are as follows:
(1) the effect of different doses of pharmaceutical composition W on BMD in osteoporotic rats is shown in table 11:
TABLE 11 BMD assay results (X + -SD) 17 weeks after treatment of Ovx rats with different dosing of pharmaceutical composition W
n is 10; p <0.05, P <0.01, P <0.001 to NS group; + -% is the percent difference compared to the NS group, (+ -) is the percent difference compared to the SHAM group (+ is increase, -is decrease)
As shown in Table 11, OVX osteoporosis control rats (NS group) showed a significant reduction in femoral (left) and lumbar (L1-L5) BMD after 17 weeks, 8.7% for the former (P <0.001) and 11.1% for the latter (P < 0.001). After oral mucosa administration of the pharmaceutical composition W for 17 weeks (6 times per week), the BMD of the femur and the lumbar vertebra of the 1390 mu g/kg group and the 4170 mu g/kg group are obviously increased (6.2-9.8%, P <0.01 or P < 0.001; 11.4-5.6%, P <0.001 or P <0.05 respectively) compared with the NS group; in addition, the increase effect of the 1390 μ g/kg group relative to the BMD of femur and lumbar vertebrae of the NS group was consistent with that of the SC group, and the increase effect of the 4170 μ g/kg group relative to the BMD of femur of the NS group was superior to that of the SC group, while the increase effect of the BMD of lumbar vertebrae was comparable to that of the SC group.
(2) Effect of different doses of pharmaceutical composition W on the biomechanical properties of osteoporotic rat bone:
TABLE 12 bone biomechanical performance measurements (X + -SD) of OVX rats treated with different doses of pharmaceutical composition W17 weeks
n is 10; p <0.05, P <0.01, P <0.001 to NS group; + -% is the percent difference compared to the NS group, (+ -) is the percent difference compared to the SHAM group (+ is increase, -is decrease)
As shown in Table 12, OVX osteoporosis control rats (NS) showed a 2.4% (P >0.05) and 32.5% (P <0.001) reduction in femoral three-point flexion maximum load and lumbar compression maximum load, respectively, over the SHAM group, when observed for a further 17 weeks. Oral mucosa administration of pharmaceutical composition W with different administration doses is adopted for treatment, and the three-point bending maximum load of femur and the compression maximum load of lumbar vertebra of 1390 mug/kg group and 4170 mug/kg group are respectively increased by 15.2% (P <0.01), 8.3, 70.9%, 61.2% (P <0.01 or P <0.001) compared with the NS group after 17 weeks; in addition, the 1390. mu.g/kg group showed a better effect of increasing the femoral three-point bending load and the lumbar compression load than the SC group, the 4170. mu.g/kg group showed a lower effect of increasing the femoral three-point bending load than the NS group, and the lumbar compression load was slightly better than the SC group.
(3) Effect of different doses of pharmaceutical composition W on serum ALP and TRAP5b activity in osteoporotic rats:
TABLE 13 serum OCN and TRAP5b assay results (X + -SD) after 17 weeks of OVX rats treated with different doses of pharmaceutical composition W
P <0.05, P <0.01, P <0.001 to NS group; + -% is the percent difference compared to the NS group, (+ -) is the percent difference compared to the SHAM group (+ is increase, -is decrease)
As shown in Table 13, OVX osteoporosis control rats (NS) were observed for a further 17 weeks, and serum Osteocalcin (OCN) restored to near SHAM group levels (+ 8.8%, P > 0.05). After oral mucosa administration treatment of the pharmaceutical composition W with different administration doses, the serum OCN level is remarkably increased compared with the NS group, wherein the 1390 mu g/kg group and the 4170 mu g/kg group are respectively increased by 131 percent and 72.4 percent (P <0.001 or P <0.05) compared with the NS group; furthermore, the OCN increasing effect of 1390. mu.g/kg group relative to NS group was the same as that of SC group, and the OCN increasing effect of 4170. mu.g/kg group relative to NS group was inferior to that of SC group. The serum anti-tartaric acid phosphatase 5b (TRAP5b) level of OVX osteoporosis control rats (NS group) is reduced by 24.7 percent compared with that of SHAM group (P <0.01), and the serum TRAP5b level is increased remarkably compared with that of NS group after oral mucosa administration treatment of pharmaceutical composition W with different concentrations, wherein 1390 mu g/kg group and 4170 mu g/kg group are respectively increased by 27.6 percent (P <0.05) and 27.5 percent (P < 0.05); furthermore, the 1390. mu.g/kg and 4170. mu.g/kg groups increased water on average slightly better than the SC group TRAP5 b.
The experimental results show that the tested drug group (ORAL) has similar bone density and bone strength compared with the normal control group (SHAM); compared with a negative control group (SC), the test drug group (ORAL, the administration dosage is 1390 mu g/kg and 4170 mu g/kg respectively) can obviously improve the bone density, the bone strength and the levels of OCN and TRAP5b in serum of the osteoporosis rats, and can also obviously improve the bone density, the bone strength and the levels of OCN and TRAP5b in serum compared with the positive control group (SC). Therefore, the pharmaceutical composition prepared in the embodiment 1 of the invention can obviously improve the osteoporosis symptoms of the ovariectomized rats, and the oral mucosa administration route realizes the good absorption and utilization of the drug rhPTH (1-34).
In animal activity experiments on the other groups of pharmaceutical compositions obtained in example 1 of the present invention, the other groups of pharmaceutical compositions a-V and X also exhibited similar activity effects to those of pharmaceutical composition W, especially the pharmaceutical composition W was the most preferred.
In conclusion, the active ingredients and the absorption promoting penetrant in the pharmaceutical composition obtained in example 1 maintain high activity, and show good penetration and absorption in penetration and absorption detection, and the pharmaceutical composition obtained in example 1 has a good prospect of being prepared into oral mucosa administration pharmaceutical preparations according to various indexes.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (7)
1. A pharmaceutical composition containing human parathyroid hormone for oral mucosa administration comprises recombinant human parathyroid hormone rhPTH (1-34), an absorption penetration enhancer, namely human hyaluronidase PH20(hPH20) and a mucosa adsorbent, wherein the concentration of the rhPTH (1-34) in the pharmaceutical composition is 100-1000 mu g/ml, and the enzymatic activity of the absorption penetration enhancer hPH20 in the pharmaceutical composition is 1000-20000U/ml; the mucosa adsorbent in the pharmaceutical composition is a composition obtained by mixing one or more than two of povidone K30, carbomer 934P and carboxymethyl chitosan in any proportion, and the concentration of the mucosa adsorbent in the pharmaceutical composition is 0.2-15 mg/ml.
2. The pharmaceutical composition containing human parathyroid hormone for oromucosal administration according to claim 1, wherein the absorption penetration enhancer is recombinant human hyaluronidase rhPH 20.
3. The pharmaceutical composition containing human parathyroid hormone for oromucosal administration according to claim 1, wherein the pharmaceutical composition further comprises sodium carboxymethyl cellulose as a thickening agent.
4. A process for the preparation of a pharmaceutical composition containing human parathyroid hormone for oromucosal administration as claimed in claim 1 or claim 3, comprising the steps of: 1) preparing a solution containing a mucoadhesive adsorbent or a solution containing a mucoadhesive adsorbent and a thickening agent; 2) adding the recombinant absorption penetration enhancer, namely the human hyaluronidase PH20 protein stock solution into the solution prepared in the step 1), and uniformly mixing; 3) adding the rhPTH (1-34) protein stock solution obtained by recombinant preparation into the mixed solution obtained in the step 2), and uniformly mixing.
5. A pharmaceutical formulation containing rhPTH (1-34) characterized by comprising the pharmaceutical composition containing human parathyroid hormone according to claim 1 or 3, for oromucosal administration.
6. The pharmaceutical formulation containing rhPTH (1-34) of claim 5, wherein said pharmaceutical formulation is a spray or a patch.
7. Use of a pharmaceutical formulation containing rhPTH (1-34) as defined in claim 5 or 6 in the manufacture of a medicament for the treatment and/or prevention of osteoporosis.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511000254 | 2015-12-28 | ||
| CN2015110002544 | 2015-12-28 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106924721A CN106924721A (en) | 2017-07-07 |
| CN106924721B true CN106924721B (en) | 2020-03-27 |
Family
ID=59444526
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611190247.XA Active CN106924721B (en) | 2015-12-28 | 2016-12-21 | Pharmaceutical composition containing human parathyroid hormone for oral mucosal administration |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106924721B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108543061A (en) * | 2018-04-11 | 2018-09-18 | 郑向鹏 | A kind of slow release tablet and preparation method thereof promoting oral cavity and/or Esophageal Mucosa reparation |
| CN110755380B (en) * | 2019-11-04 | 2021-09-24 | 中国人民解放军军事科学院军事医学研究院 | Parathyroid hormone 1-34 nasal spray, preparation method and application thereof |
| CN113967249A (en) * | 2021-12-10 | 2022-01-25 | 深圳先进技术研究院 | Application of parathyroid hormone in preparation of medicine or health-care product for treating male depression |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000038732A1 (en) * | 1998-12-23 | 2000-07-06 | Esparma Gmbh | Hyaluronate lyase used for promoting penetration in topical agents |
| CN101843595A (en) * | 2009-07-16 | 2010-09-29 | 中国人民解放军第四军医大学 | Formula of oral mucosa absorbing dosage form of medicines of polypeptides and proteins, and preparation method and application thereof |
| CN103468662A (en) * | 2013-09-29 | 2013-12-25 | 惠觅宙 | Recombined human hyaluronidase, production and purification method and preparations thereof, use method and application |
| US8795654B2 (en) * | 2007-06-19 | 2014-08-05 | Tamara P. Uvarkina | Methods for improving the tissue penetration of a drug by administering the drug together with a hyaluronidase protein |
-
2016
- 2016-12-21 CN CN201611190247.XA patent/CN106924721B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000038732A1 (en) * | 1998-12-23 | 2000-07-06 | Esparma Gmbh | Hyaluronate lyase used for promoting penetration in topical agents |
| US8795654B2 (en) * | 2007-06-19 | 2014-08-05 | Tamara P. Uvarkina | Methods for improving the tissue penetration of a drug by administering the drug together with a hyaluronidase protein |
| CN101843595A (en) * | 2009-07-16 | 2010-09-29 | 中国人民解放军第四军医大学 | Formula of oral mucosa absorbing dosage form of medicines of polypeptides and proteins, and preparation method and application thereof |
| CN103468662A (en) * | 2013-09-29 | 2013-12-25 | 惠觅宙 | Recombined human hyaluronidase, production and purification method and preparations thereof, use method and application |
Non-Patent Citations (3)
| Title |
|---|
| Clinical pharmacology and possible applications of hyaluronidase with reference to Hylase "Dessau";Farr C等;《Winner Medizinische Wochenschrift》;19970228;第147卷(第15期);第347-355页 * |
| The Effects of Hyaluronidase on the Efficacy and on the Pain of Administration of 1% Lidocaine;Nevarre D R等;《Plastic and Reconstructive Surgery》;19980228;第101卷(第2期);第365-369页 * |
| 透明质酸酶促进胰岛素经口腔粘膜吸收的初步探讨;丁明星等;《临床医药文献杂志》;20150831;第2卷(第23期);摘要、第4721页右栏第1段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106924721A (en) | 2017-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| RU2493868C2 (en) | Pharmaceutical compositions containing hgh for oral administration | |
| JP2021520391A (en) | Pharmaceutical compositions for transmucosal delivery of therapeutic peptides and proteins | |
| JP5611033B2 (en) | Composition for transmucosal delivery of polypeptides | |
| US20130034597A1 (en) | Orally bioavailable peptide drug compositions and methods thereof | |
| EA031134B1 (en) | Rapid-acting insulin compositions | |
| CN101151048A (en) | Compositions and methods for enhanced mucosal delivery of parathyroid hormone | |
| KR101820115B1 (en) | Stabilization of fsh | |
| CN106924721B (en) | Pharmaceutical composition containing human parathyroid hormone for oral mucosal administration | |
| KR101940341B1 (en) | Pharmaceutical composition | |
| CN113398250B (en) | Liquid gonadotrophin medicinal preparation and use | |
| JP4699991B2 (en) | Liquid pharmaceutical composition of FSH and LH with non-ionic surfactant | |
| JP2016523267A (en) | Heprapeptide formulation | |
| JP6869255B2 (en) | Frozen pharmaceutical product and its use | |
| JP2015038111A (en) | Liquid formulation of g-csf | |
| JP2007161702A (en) | Pharmaceutical composition for aqueous inhalation | |
| US20210251886A1 (en) | Oral mucosal delivery systems comprising monophasic concentrate of teriparatide | |
| CN106924723B (en) | Pharmaceutical composition containing human glucagon-like peptide-1 analogue administered through oral mucosa | |
| JP4283776B2 (en) | HGH-containing pharmaceutical composition | |
| CN115884783A (en) | Low dose pharmaceutical compositions of GHRH analogs and uses thereof | |
| JP2024515392A (en) | Pharmaceutical preparations containing cyclodextrin | |
| WO2023110833A1 (en) | Pharmaceutical formulations comprising a cyclodextrin | |
| JP2009221159A (en) | Medicinal preparation containing pegylated lactoferrin as main component |
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 | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 610041 South Keyuan Road, Chengdu High-tech Zone, Chengdu, Sichuan Province, No. 88, 10 buildings, 2 floors, 203 Applicant after: Xinlitai (Chengdu) Biotechnology Co., Ltd. Address before: 610041 South Keyuan Road, Chengdu High-tech Zone, Chengdu, Sichuan Province, No. 88, 10 buildings, 2 floors, 203 Applicant before: Chengdu Jinkai Biological Technology Co., Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP02 | Change in the address of a patent holder | ||
| CP02 | Change in the address of a patent holder |
Address after: 610093 No. 1, floor 3, building 4, No. 1, Keyuan South Road, high tech Zone, Chengdu, Sichuan Patentee after: Xinlitai (Chengdu) Biotechnology Co.,Ltd. Address before: Room 203, floor 2, building 10, No. 88, Keyuan South Road, Chengdu hi tech Zone, Chengdu, Sichuan 610041 Patentee before: Xinlitai (Chengdu) Biotechnology Co.,Ltd. |