DE4026146A1 - Compsn. for treating osteoporosis without affecting calcium balance - contains central region of parathormone and biologically active vitamin=D metabolite - Google Patents
Compsn. for treating osteoporosis without affecting calcium balance - contains central region of parathormone and biologically active vitamin=D metaboliteInfo
- Publication number
- DE4026146A1 DE4026146A1 DE19904026146 DE4026146A DE4026146A1 DE 4026146 A1 DE4026146 A1 DE 4026146A1 DE 19904026146 DE19904026146 DE 19904026146 DE 4026146 A DE4026146 A DE 4026146A DE 4026146 A1 DE4026146 A1 DE 4026146A1
- Authority
- DE
- Germany
- Prior art keywords
- hpth
- metabolite
- biologically active
- amino acids
- active vitamin
- 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.)
- Ceased
Links
- 239000002207 metabolite Substances 0.000 title claims abstract 6
- 239000011575 calcium Substances 0.000 title abstract description 21
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 title abstract description 20
- 229910052791 calcium Inorganic materials 0.000 title abstract description 20
- 102000003982 Parathyroid hormone Human genes 0.000 title abstract description 16
- 108090000445 Parathyroid hormone Proteins 0.000 title abstract description 16
- 239000000199 parathyroid hormone Substances 0.000 title abstract description 13
- 208000001132 Osteoporosis Diseases 0.000 title abstract description 4
- 239000011782 vitamin Substances 0.000 title 1
- 229940088594 vitamin Drugs 0.000 title 1
- 229930003231 vitamin Natural products 0.000 title 1
- 235000013343 vitamin Nutrition 0.000 title 1
- 150000003722 vitamin derivatives Chemical class 0.000 title 1
- 150000001413 amino acids Chemical class 0.000 claims abstract description 9
- FCKJYANJHNLEEP-XRWYNYHCSA-N (24R)-24,25-dihydroxycalciol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CC[C@@H](O)C(C)(C)O)C)=C\C=C1\C[C@@H](O)CCC1=C FCKJYANJHNLEEP-XRWYNYHCSA-N 0.000 claims abstract description 7
- 230000001225 therapeutic effect Effects 0.000 claims abstract description 6
- JWUBBDSIWDLEOM-NQZHSCJISA-N 25-hydroxy-3 epi cholecalciferol Chemical compound C1([C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CCCC(C)(C)O)C)=CC=C1C[C@H](O)CCC1=C JWUBBDSIWDLEOM-NQZHSCJISA-N 0.000 claims abstract 5
- 230000004048 modification Effects 0.000 claims abstract 4
- 238000012986 modification Methods 0.000 claims abstract 4
- 102000037865 fusion proteins Human genes 0.000 claims abstract 2
- 108020001507 fusion proteins Proteins 0.000 claims abstract 2
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 claims description 11
- 229930003316 Vitamin D Natural products 0.000 claims description 6
- 235000019166 vitamin D Nutrition 0.000 claims description 6
- 239000011710 vitamin D Substances 0.000 claims description 6
- 229940046008 vitamin d Drugs 0.000 claims description 6
- 150000003710 vitamin D derivatives Chemical class 0.000 claims description 5
- 230000010072 bone remodeling Effects 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 abstract description 2
- 230000002195 synergetic effect Effects 0.000 abstract description 2
- 210000004899 c-terminal region Anatomy 0.000 abstract 2
- 108010060933 parathyroid hormone (28-48) Proteins 0.000 abstract 1
- 229960001319 parathyroid hormone Drugs 0.000 description 11
- GMRQFYUYWCNGIN-ZVUFCXRFSA-N 1,25-dihydroxy vitamin D3 Chemical compound C1([C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CCCC(C)(C)O)C)=CC=C1C[C@@H](O)C[C@H](O)C1=C GMRQFYUYWCNGIN-ZVUFCXRFSA-N 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 210000000988 bone and bone Anatomy 0.000 description 7
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 7
- 239000012634 fragment Substances 0.000 description 6
- 210000000056 organ Anatomy 0.000 description 6
- 230000009102 absorption Effects 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 5
- 238000010348 incorporation Methods 0.000 description 5
- 235000005282 vitamin D3 Nutrition 0.000 description 5
- 239000011647 vitamin D3 Substances 0.000 description 5
- 229940021056 vitamin d3 Drugs 0.000 description 5
- 241000287828 Gallus gallus Species 0.000 description 4
- 235000013330 chicken meat Nutrition 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000001925 catabolic effect Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- GMRQFYUYWCNGIN-UHFFFAOYSA-N 1,25-Dihydroxy-vitamin D3' Natural products C1CCC2(C)C(C(CCCC(C)(C)O)C)CCC2C1=CC=C1CC(O)CC(O)C1=C GMRQFYUYWCNGIN-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 230000037176 bone building Effects 0.000 description 2
- GMRQFYUYWCNGIN-NKMMMXOESA-N calcitriol Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@@H](CCCC(C)(C)O)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C GMRQFYUYWCNGIN-NKMMMXOESA-N 0.000 description 2
- 210000002257 embryonic structure Anatomy 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 230000002297 mitogenic effect Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 102000004420 Creatine Kinase Human genes 0.000 description 1
- 108010042126 Creatine kinase Proteins 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 102000030621 adenylate cyclase Human genes 0.000 description 1
- 108060000200 adenylate cyclase Proteins 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229940072107 ascorbate Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000002449 bone cell Anatomy 0.000 description 1
- 230000002308 calcification Effects 0.000 description 1
- 235000020964 calcitriol Nutrition 0.000 description 1
- 239000011612 calcitriol Substances 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000000668 effect on calcium Effects 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 210000004349 growth plate Anatomy 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000001872 metatarsal bone Anatomy 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000009103 reabsorption Effects 0.000 description 1
- 230000027425 release of sequestered calcium ion into cytosol Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- -1 vitamin D compounds Chemical class 0.000 description 1
- 239000005544 vitamin D3 metabolite Substances 0.000 description 1
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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Endocrinology (AREA)
- Immunology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
Parathormon (PTH) reguliert den Calciumspiegel des Blutplasmas, indem es u. a. auf- und abbauende Wirkungen auf das Skelettsystem entfaltet. An der Niere fördert das Hormon die Rückresorption von Calcium. Vitamin D₃ spielt ebenfalls eine wichtige Rolle in der Calcium- und Phosphat-Homöostase. Die beiden in der Niere gebildeten Vitamin D₃-Metabolita 1α,25-Dihydroxyvitamin D₃ und 24,25-Dihydroxyvitamin D₃ haben einen aktiven biologischen Einfluß auf den Knochenumbau.Parathyroid hormone (PTH) regulates the calcium level in the blood plasma, by u. a. build-up and breakdown effects on the skeletal system unfolded. In the kidney, the hormone promotes reabsorption of Calcium. Vitamin D₃ also plays an important role in Calcium and phosphate homeostasis. The two in the kidney formed vitamin D₃ metabolita 1α, 25-dihydroxyvitamin D₃ and 24,25-Dihydroxyvitamin D₃ have an active biological influence on the bone remodeling.
Das "bone remodelling" im Knochen beruht auf einem Gleichgewicht von Neubildung und Resorption. Über die Knochenneubildung und die damit verbundenen physiologischen Faktoren ist wenig bekannt, im Gegensatz zu dem Einfluß von z. B. Parathormon und aktiven Metaboliten des Vitamin D₃ auf die Resorption. Generell wurden in der Vergangenheit nahezu alle biologischen Effekte des PTH seinen aminoterminalen 34 Aminosäuren zugeschrieben, und für die meisten seiner Aktivitäten cAMP als Signalüberträger identifiziert (Adv. Protein Chem., 35 (1982) 323-396; Potts et al.). Tatsächlich ist die Stimulation der cAMP-Synthese offenbar für seine knochenabbauende, calciumfreisetzende Funktion essentiell (Science, 197 (1977) 663-665; Wong et al.). Auch für Vitamin D₃ sind vielfach eher katabole Effekte berichtet worden. Z. B. wird in Science, 175 (1972) 768-769 (Hölick & De Luca) die Calcium- Resorption durch 1,25(OH)₂D₃ an Knochenorgankulturen von foetalen Ratten beschrieben. Sowohl 1,25(OH)₂D₃ als auch Parathormon setzen die Aktivität der alkalischen Phosphatase in Osteoblasten herab, die für Calcifizierungsprozesse wesentlich ist (Science, 197 (1977) 663-665; Wong et al.).The bone remodeling in the bone is based on a balance of new formation and absorption. About new bone formation and little is known about the associated physiological factors Contrary to the influence of z. B. parathyroid hormone and active Metabolites of vitamin D₃ on absorption. Generally, in the past almost all biological effects of PTH amino terminal attributed to 34 amino acids, and for most of its activities identified cAMP as a signal transmitter (Adv. Protein Chem., 35 (1982) 323-396; Potts et al.). Indeed it is the stimulation of cAMP synthesis apparently for his Bone-depleting, calcium-releasing function essential (Science, 197 (1977) 663-665; Wong et al.). Also for vitamin D₃ catabolic effects have often been reported. For example, in Science, 175 (1972) 768-769 (Hölick & De Luca) the calcium Absorption by 1,25 (OH) ₂D₃ on bone organ cultures of fetal Rats described. Set both 1,25 (OH) ₂D₃ and parathyroid hormone decrease the activity of alkaline phosphatase in osteoblasts, which is essential for calcification processes (Science, 197 (1977) 663-665; Wong et al.).
Scheinbar in Abhängigkeit vom Alter des Zielorgans kann 1,25(OH)₂D₃ aber auch die katabole PTH-Wirkung auf Maus-Calvarien unterdrücken (Mechanism of Ageing and Development, 49 (1989) 199- 209; Goldhaber & Rabadjÿa): Die durch PTH induzierte Calciumfreisetzung wird durch Zugabe von 1,25(OH)₂D₃ zu 75 Tage alten Calvarien inhibiert, während es die durch 5 Tage alte Organe kaum signifikant beeinflußt. Offensichtlich unterdrückt Vitamin D die cAMP-Antwort der Zielzellen auf aminoterminal intakte PTH- Peptide (Calcif. Tissue Int., 46 (1990) 339-345; Mortensen et al.) über eine Beeinträchtigung des als Kopplung zwischen PTH-Rezeptor und Adenylat-Cyclase dienenden Gs-Proteins (Biochem. Pharmocol. 38 (1989) 3111-3114, de Cremoux et al.; Biochem. Biophys. Res. Commun. 168 (1990) 889-897, Ikeda et al.).Apparently depending on the age of the target organ, 1,25 (OH) ₂D₃ can also suppress the catabolic PTH effect on mouse calvaria (Mechanism of Aging and Development, 49 (1989) 199-209; Goldhaber & Rabadjÿa): The PTH induced calcium release is inhibited by adding 1,25 (OH) ₂D₃ to 75-day-old calvaria, while it hardly influences the organs by 5-day-old. Vitamin D apparently suppresses the cAMP response of the target cells to amino terminally intact PTH peptides (Calcif. Tissue Int., 46 (1990) 339-345; Mortensen et al.) By impairing the coupling between the PTH receptor and adenylate cyclase serving G s proteins (Biochem. Pharmocol. 38 (1989) 3111-3114, de Cremoux et al .; Biochem. Biophys. Res. Commun. 168 (1990) 889-897, Ikeda et al.).
Eine synergistische knochenaufbauende Wirkung durch die Kombination von N-terminal offenbar intaktem PTH+1α,25(OH)₂D₃ und 24R,25(OH)₂D₃ wurde an Femura von 9 Tage alten Hühnerembryonen beobachtet (Nature, 286 (1980) 262-264; Endo et al.). Von therapeutischem Nutzen zur Behandlung männlicher Osteoporose- Patienten erwies sich auch eine Kombination von hPTH(1-34) mit 1,25(OH)₂D₃ (J. Bone Mineral Res., 1 (1986) 377-381; Slovik et al.).A synergistic bone-building effect through the combination of N-terminally intact PTH + 1α, 25 (OH) ₂D₃ and 24R, 25 (OH) ₂D₃ was observed in the femur of 9-day-old chicken embryos (Nature, 286 (1980) 262-264; Endo et al.). A combination of hPTH ( 1-34 ) with 1,25 (OH) ₂D₃ (J. Bone Mineral Res., 1 (1986) 377-381; Slovik et al.) Also proved to be of therapeutic benefit for the treatment of male osteoporosis patients. .
In einer vorangegangenen Patentanmeldung wurde der mitogene Effekt mittregionaler PTH-Fragmente, z. B. des hPTH(28-48), beschrieben (P 37 25 319.0; s. außerdem J. Biol. Chem., 264 (1989) 11087-11092; Schlüter et al.). Derartige Fragmente bewirken u. a. eine gesteigerte Proliferation von Chondrocyten embryonaler Hühner, erhöhte DNA-Synthese und Creatinkinase-Aktivität in Calvarienzellen (in vitro) oder epiphsealen Knorpel- sowie diaphysealen Knochenzellen der Ratte (in vivo) (s. Anhang zur Patentanmeldung P 37 25 319.0).In a previous patent application, the mitogenic effect of mid-regional PTH fragments, e.g. B. of hPTH ( 28-48 ) (P 37 25 319.0; see also J. Biol. Chem., 264 (1989) 11087-11092; Schlueter et al.). Such fragments cause, inter alia, an increased proliferation of chondrocytes of embryonic chickens, increased DNA synthesis and creatine kinase activity in calvary cells (in vitro) or epiphseal cartilage and diaphyseal bone cells of the rat (in vivo) (see appendix to patent application P 37 25 319.0) .
Neben der Zellproliferation knochenaufbauender Zellen wäre jedoch der gesteigerte Einbau von Calcium in das Skelettgerüst ein speziell bei Osteoporose erwünschter Effekt. Zu seinem Nachweis wird ein Organkultur-Testsystem aus Metatarsalia embryonaler Hühner verwendet. In diesem Testsystem läßt sich der katabole calciummobilisierende Effekt aminoterminal intakter PTH-Peptide, z. B. des hPTH(1-34), darstellen (s. Fig. 1A). Im Gegensatz dazu hat sich jedoch gezeigt, daß mittregionale, mitogen wirkende Fragmente keinen Einfluß auf die Calciumbilanz besitzen (Fig. 1B), obwohl sie in derselben Kultur eine Zunahme des Wachstums des epiphysealen Knorpels bewirken (nicht gezeigt).In addition to the cell proliferation of bone-building cells, the increased incorporation of calcium into the skeletal structure would be a particularly desirable effect in osteoporosis. An organ culture test system from Metatarsalia embryonic chickens is used to detect it. In this test system, the catabolic calcium mobilizing effect of amino terminal intact PTH peptides, e.g. B. the hPTH ( 1-34 ), (see Fig. 1A). In contrast, however, it has been shown that mid-regional, mitogenic fragments have no influence on the calcium balance ( FIG. 1B), although they cause an increase in the growth of the epiphyseal cartilage in the same culture (not shown).
In Kombination mit aminoterminal intakten PTH-Peptiden, die die cAMP-Synthese stimulieren (z. B. hPTH(1-34)), unterdrückt Vitamin D₃ auch in dem hier verwendeten Testsystem die Stimulation der Calciumfreisetzung durch hPTH(1-34) (Fig. 2). Überraschenderweise führt jedoch die Kombination für sich inaktiver mittregionaler Fragmente mit den geeigneten Vitamin-D₃-Metaboliten zu einem beträchtlichen Einbau an Calcium (Fig. 3). Die Vitamin-D- Verbindungen selbst besitzen keine oder nur schwache Aktivität. Die so behandelten Knochen weisen eine der Abnahme im Kulturüberstand entsprechende Aufnahme von Calcium auf.In combination with amino terminally intact PTH peptides that stimulate cAMP synthesis (e.g. hPTH ( 1-34 )), vitamin D₃ also suppresses the stimulation of calcium release by hPTH ( 1-34 ) in the test system used here ( Fig . 2). Surprisingly, however, the combination of inactive central region fragments with the appropriate vitamin D₃ metabolites leads to a considerable incorporation of calcium ( Fig. 3). The vitamin D compounds themselves have little or no activity. The bones treated in this way show an absorption of calcium corresponding to the decrease in the culture supernatant.
Die erfindungsgemäße Wirkung wurde an einem Knochenorgankulturmodell gezeigt. Aus 17 Tage alten Hühnerembryonen wurden die Metatarsalia isoliert und in 4 ml FCS- freiem DME Medium kultiviert. Alle 2 Tage wurde frisches Ascorbat zugegeben. Die Organkulturen wurden nach einer Adaptionsphase von 24 Stunden mit den PTH Fragmenten bzw. Kombination mit 1,25- Dihydroxycholecalciferol induziert.The effect according to the invention was in one Bone organ culture model shown. From 17 days old Chicken embryos, the metatarsals were isolated and placed in 4 ml FCS free DME medium cultivated. Fresh ascorbate was given every 2 days admitted. The organ cultures were after an adaptation phase of 24 hours with the PTH fragments or combination with 1,25- Dihydroxycholecalciferol induced.
Zur Bestimmung des Einbaus bzw. der Mobilisierung von Calcium durch die Knochen wurden täglich Proben entnommen und der Calciumgehalt des Mediums mit einem Atomabsorptionsspektrometer bestimmt.To determine the installation or mobilization of calcium samples were taken daily through the bones and the Calcium content of the medium with an atomic absorption spectrometer certainly.
Die hier gezeigten Ergebnisse zeigen jeweils die innerhalb von ca. 145 Stunden nach Induktion gemessenen Veränderungen des Calciumgehalts im Kulturüberstand von Metatarsalia, die (A) mit 1×10-8 M hPTH(1-34) oder (B) mit 1×10-8 M hPTH(28-48) behandelt worden waren. Während hPTH(1-34) zu einem signifikanten Anstieg des Calciumgehaltes im Kulturüberstand und somit zu einer Calciumfreisetzung aus den Knochen führt, ist hPTH(28-48) in dieser Hinsicht völlig inaktiv. The results shown here each show the changes in the calcium content in the culture supernatant of metatarsalia measured within approx. 145 hours after induction, the (A) with 1 × 10 -8 M hPTH ( 1-34 ) or (B) with 1 × 10 - 8 M hPTH ( 28-48 ) had been treated. While hPTH ( 1-34 ) leads to a significant increase in the calcium content in the culture supernatant and thus to a calcium release from the bones, hPTH ( 28-48 ) is completely inactive in this regard.
Die durch 10-8 M hPTH(1-34) erzielte Calciummobilisierung (s. mit (1-34) bezeichnete Säule; K: unbehandelte Kontrollkultur) kann durch Kombination mit verschiedenen Konzentrationen an 1,25(OH)₂D₃ (1×10-12 M (bezeichnet als 1E-12M) bis 1×10-8 M (1E-8M)) signifikant unterdrückt werden. SEM: Standardfehler des Mittelwertes.The calcium mobilization achieved by 10 -8 M hPTH ( 1-34 ) (see column labeled ( 1-34 ); K: untreated control culture) can be combined with various concentrations of 1,25 (OH) ₂D₃ (1 × 10 - 12 M (referred to as 1E-12M) to 1 × 10 -8 M (1E-8M)) were significantly suppressed. SEM: standard error of the mean.
Das mittregionale Fragment hTPH(28-48) zeigt bei alleiniger Induktion keine Wirkung auf Calciummobilisierung oder -einbau (s. mit (28-48) bezeichnete Säule; K: unbehandelte Kontrollkultur). In Kombination mit verschiedenen Konzentrationen an 1,25(OH)₂D₃ (1×10-13 M) (bezeichnet als 1E-13M) bis 1×10-8 M (1E-8M)) läßt sich ein signifikanter Calciumeinbau nach ca. 145 Stunden in Kultur beobachten: SEM: Standardfehler des Mittelwertes.The central region fragment hTPH ( 28-48 ) shows no effect on calcium mobilization or incorporation when induction alone (see column labeled ( 28-48 ); K: untreated control culture). In combination with different concentrations of 1,25 (OH) ₂D₃ (1 × 10-13 M) (referred to as 1E-13M) to 1 × 10 -8 M (1E-8M)), significant calcium incorporation can be achieved after approx. 145 Observe hours in culture: SEM: standard error of the mean.
Eine Vitamin D₃ Kontrolle mit einer Konzentration von 10-8 M zeigt einen sehr schwachen Einbau um die 1,24 µg Ca (SEM=0,30) (nicht gezeigt).A vitamin D₃ control with a concentration of 10 -8 M shows a very weak incorporation around 1.24 µg Ca (SEM = 0.30) (not shown).
Claims (4)
- a) eine pPTH-, bPTH- oder hPTH-Modifikation, bei der der natürliche N-Terminus um ein bis 29 Aminosäuren deletiert ist und der natürliche C-Terminus um ein bis 50 Aminosäuren deletiert sein kann (wie z. B. in hPTH(28-48)), und
- b) mindestens einen biologisch aktiven Vitamin-D-Metaboliten (z. B. 1,25- oder 24,25-Dihydroxyvitamin D₃).
- a) a pPTH, bPTH or hPTH modification in which the natural N-terminus is deleted by one to 29 amino acids and the natural C-terminus can be deleted by one to 50 amino acids (such as in hPTH ( 28-48 )), and
- b) at least one biologically active vitamin D metabolite (z. B. 1,25- or 24,25-dihydroxyvitamin D₃).
- a) eine pPTH-, bPTH- oder hPTH-Modifikation, bei der der natürliche N-Terminus um ein bis 1000 Aminosäuren verlängert ist und der natürliche C-Terminus um ein bis 50 Aminosäuren deletiert sein kann (wie z. B. in Pro(-1)-hPTH(1-84) oder Cro- β-Galaktosidase-hPTH-Fusionsproteinen gemäß P 37 25 320)), und
- b) mindestens einen biologisch aktiven Vitamin-D-Metaboliten (z. B. 1,25- oder 24,25-Dihydroxyvitamin D₃).
- a) a pPTH, bPTH or hPTH modification in which the natural N-terminus is extended by one to 1000 amino acids and the natural C-terminus can be deleted by one to 50 amino acids (as e.g. in Pro ( -1) -hPTH ( 1-84 ) or Cro-β-galactosidase-hPTH fusion proteins according to P 37 25 320)), and
- b) at least one biologically active vitamin D metabolite (z. B. 1,25- or 24,25-dihydroxyvitamin D₃).
- a) pPTH-, bPTH- oder hPTH(1-84), das mit Hilfe des in P 37 25 320 beschriebenen Expressionssystems hergestellt wurde und
- b) mindestens einen biologisch aktiven Vitamin-D-Metaboliten (z. B. 1,25- oder 24,25-Dihydroxyvitamin D₃).
- a) pPTH, bPTH or hPTH ( 1-84 ), which was produced with the help of the expression system described in P 37 25 320 and
- b) at least one biologically active vitamin D metabolite (z. B. 1,25- or 24,25-dihydroxyvitamin D₃).
- a) eine Variante von pPTH-, bPTH- oder hPTH(1-84) gemäß P 39 05 606 und
- b) mindestens einen biologisch aktiven Vitamin-d-Metaboliten (z. B. 1,25- oder 24,25-Dihydroxyvitamin D₃).
- a) a variant of pPTH, bPTH or hPTH ( 1-84 ) according to P 39 05 606 and
- b) at least one biologically active vitamin D metabolite (e.g. 1,25- or 24,25-dihydroxyvitamin D₃).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19904026146 DE4026146A1 (en) | 1990-08-17 | 1990-08-17 | Compsn. for treating osteoporosis without affecting calcium balance - contains central region of parathormone and biologically active vitamin=D metabolite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19904026146 DE4026146A1 (en) | 1990-08-17 | 1990-08-17 | Compsn. for treating osteoporosis without affecting calcium balance - contains central region of parathormone and biologically active vitamin=D metabolite |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DE4026146A1 true DE4026146A1 (en) | 1992-02-20 |
Family
ID=6412462
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE19904026146 Ceased DE4026146A1 (en) | 1990-08-17 | 1990-08-17 | Compsn. for treating osteoporosis without affecting calcium balance - contains central region of parathormone and biologically active vitamin=D metabolite |
Country Status (1)
| Country | Link |
|---|---|
| DE (1) | DE4026146A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1534309A2 (en) * | 2002-08-09 | 2005-06-01 | Scantibodies Laboratory, Inc. | Parathyroid hormone antagonists and uses thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0197514A1 (en) * | 1985-04-04 | 1986-10-15 | The General Hospital Corporation | Pharmaceutical composition for use in increasing bone mass |
| DE3725319A1 (en) * | 1987-07-30 | 1989-02-09 | Biotechnolog Forschung Gmbh | PTH MODIFICATIONS AND THERAPEUTIC COMPOSITIONS THAT CONTAIN AND MAKE IT OUT |
-
1990
- 1990-08-17 DE DE19904026146 patent/DE4026146A1/en not_active Ceased
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0197514A1 (en) * | 1985-04-04 | 1986-10-15 | The General Hospital Corporation | Pharmaceutical composition for use in increasing bone mass |
| DE3725319A1 (en) * | 1987-07-30 | 1989-02-09 | Biotechnolog Forschung Gmbh | PTH MODIFICATIONS AND THERAPEUTIC COMPOSITIONS THAT CONTAIN AND MAKE IT OUT |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1534309A2 (en) * | 2002-08-09 | 2005-06-01 | Scantibodies Laboratory, Inc. | Parathyroid hormone antagonists and uses thereof |
| EP1534309A4 (en) * | 2002-08-09 | 2009-11-11 | Scantibodies Lab Inc | Parathyroid hormone antagonists and uses thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE4130545C2 (en) | Use of a bone growth promoting composition | |
| Weinstein et al. | Bone histomorphometry in vitamin D-deficient rats infused with calcium and phosphorus | |
| DE69423146T2 (en) | Parathyroid hormone and raloxifene to increase bone mass | |
| DE3686343T2 (en) | PHARMACEUTICAL PREPARATION FOR APPLICATION FOR BONE MEASUREMENT. | |
| DE3687899T2 (en) | METHOD AND COMPOSITION FOR REGULATING THE SERUM CALCIUM LEVEL OF MAMMALS. | |
| Yeh et al. | Effect of growth hormone administration and treadmill exercise on serum and skeletal IGF-I in rats | |
| DE69734157T2 (en) | PARATHYROIDHORMONE-RELATED PEPTIDE ANALOG | |
| DE3851776T2 (en) | Use of IGF-II for the treatment of bone diseases. | |
| Hallick et al. | Metabolites of dihydrotachysterol3 in target tissues | |
| DE69826132T2 (en) | PHARMACEUTICAL COMBINATION PREPARATION FROM PARATHORMONE AND AN INHIBITOR OF BONE RESORPTION | |
| WO1999032620A9 (en) | Insulin-like growth factor binding protein fragments and the utilization thereof | |
| WO1991006564A1 (en) | hPTH (1-37) FRAGMENT, ITS PRODUCTION, DRUG CONTAINING IT AND ITS USE | |
| Ma et al. | Prostaglaridin E2 adds bone to a cancellous bone site with a closed growth plate and low bone turnover in ovariectomized rats | |
| Hanssen et al. | Regulation of secretion of the teleost fish hormone stanniocalcin: effects of extracellular calcium | |
| DE69000640T2 (en) | STABILIZED CALCITON PREPARATION. | |
| DE69026986T2 (en) | Amylin for the treatment of bone disorders | |
| Price et al. | Concurrent warfarin treatment further reduces bone mineral levels in 1, 25-dihydroxyvitamin D3-treated rats. | |
| Mohan et al. | Serum insulin-like growth factor binding protein (IGFBP)-4 and IGFBP-5 levels in aging and age-associated diseases | |
| DE4026146A1 (en) | Compsn. for treating osteoporosis without affecting calcium balance - contains central region of parathormone and biologically active vitamin=D metabolite | |
| DE3243358C2 (en) | ||
| DE69133074T2 (en) | Agent for treating bone diseases containing a basic fibroblast growth factor | |
| DE69121964T2 (en) | Growth-inhibiting factor and cDNA coding for the growth-inhibiting factor | |
| DE4215534C1 (en) | Solns. used to treat bee diseases - contg. lactic acid, aminoacid(s) and iron, manganese and phosphate salts | |
| EP1156811B1 (en) | Copper agonist that binds on the copper binding site of amyloid precursor protein (app) and/or exerts an inhibiting effect on the release of amyloid-beta-peptide | |
| EP0301484B1 (en) | PTH modifications and therapeutic compositions containing them or consisting of them |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| OP8 | Request for examination as to paragraph 44 patent law | ||
| 8131 | Rejection |