Discovery of a non-peptide small molecule that selectively mimics the biological actions of calcitonin

Katayama, Toyoko; Furuya, Mayumi; Yamaichi, Kozo; Konishi, Kyoko; Sugiura, Namino; Murafuji, Hidenobu; Magota, Koji; Saito, Masayuki; Tanaka, Shoji; Oikawa, Shinzo

doi:10.1016/s0304-4165(01)00125-8

Cited by 20 publications

(14 citation statements)

References 26 publications

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“…Salmon calcitonin binds to the CT receptor of the T47D cells, which belongs to a subfamily of GTP-binding (G)-protein-coupled receptors, activating G-proteins that in turn induce the activation of adenylyl cyclase and phospholipase C to generate the intracellular second messenger molecules, cyclic adenosine 3P,5P-monophosphate (cAMP) (Katayama et al, 2001). Studies by Moe and Kaiser (1985); Epand et al (1986) and Andreotti et al (2006), have shown the importance of maintaining the secondary structure of sCT (the a-helix), in order to have a fully cAMPsecreting activity (biological activity).…”

Section: Discussionmentioning

confidence: 99%

Formulation, stability and pharmacokinetics of sugar-based salmon calcitonin-loaded nanoporous/nanoparticulate microparticles (NPMPs) for inhalation

Amaro

Tewes

Gobbo

et al. 2015

International Journal of Pharmaceutics

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A challenge exists to produce dry powder inhaler (DPI) formulations with appropriate formulation stability, biological activity and suitable physicochemical and aerosolisation characteristics that provide a viable alternative to parenteral formulations. The present study aimed to produce sugar-based nanoporous/nanoparticulate microparticles (NPMPs) loaded with a therapeutic peptide - salmon calcitonin (sCT). The physicochemical properties of the powders and their suitability for pulmonary delivery of sCT were determined. Production of powders composed of sCT loaded into raffinose or trehalose with or without hydroxypropyl-β-cyclodextrin was carried out using a laboratory scale spray dryer. Spray dried microparticles were spherical, porous and of small geometric size (≤2 μm). Aerodynamic assessment showed that the fine particle fraction (FPF) less than 5 μm ranged from 45 to 86%, depending on the formulation. The mass median aerodynamic diameter (MMAD) varied between 1.9 and 4.7 μm. Compared to unprocessed sCT, sCT:raffinose composite systems presented a bioactivity of approximately 100% and sCT:trehalose composite systems between 70-90% after spray drying. Storage stability studies demonstrated composite systems with raffinose to be more stable than those containing trehalose. These sugar-based salmon calcitonin-loaded NPMPs retain reasonable sCT bioactivity and have micromeritic and physicochemical properties which indicate their suitability for pulmonary delivery. Formulations presented a similar pharmacokinetic profile to sCT solution. Hence the advantage of a dry powder formulation is its non-invasive delivery route and ease of administration of the sCT.

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Section: Discussionmentioning

confidence: 99%

Formulation, stability and pharmacokinetics of sugar-based salmon calcitonin-loaded nanoporous/nanoparticulate microparticles (NPMPs) for inhalation

Amaro

Tewes

Gobbo

et al. 2015

International Journal of Pharmaceutics

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“…Family B GPCRs contain many important drug targets, but development of small molecules targeting this family has lagged far behind the Family A members. Currently, a few such molecules are available for receptors for corticotrophin-releasing factor (24,25), glucagon (26,27), glucagon-like peptide 1 (28 -30), CT (12)(13)(14), and CT gene-related peptide (31). However, the molecular mechanism of actions for these molecules remains unknown.…”

Section: Discussionmentioning

confidence: 99%

“…For the CT receptor, several such ligands have been developed (12)(13)(14). However, the mechanisms of their actions at their receptors remain unclear.…”

Section: Calcitonin (Ct)mentioning

confidence: 99%

Juxtamembranous Region of the Amino Terminus of the Family B G Protein-coupled Calcitonin Receptor Plays a Critical Role in Small-molecule Agonist Action

Dong

Cox

Miller

2009

Journal of Biological Chemistry

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The Family B G protein-coupled calcitonin receptor is an important drug target. The aim of this work was to elucidate the molecular mechanism of action of small-molecule agonist ligands acting at this receptor, comparing it with the action mechanism of the receptor's natural peptide ligand. cAMP responses to four non-peptidyl ligands and calcitonin were studied in COS-1 cells expressing wild-type and chimeric calcitoninsecretin receptors. All compounds were full agonists at the calcitonin receptor with no activity at the secretin receptor. Only chimeric constructs including the calcitonin receptor amino terminus exhibited responses to any of these ligands. We progressively truncated this domain and tested constructs for cAMP responses. Although calcitonin was able to activate the calcitonin receptor fully with the first 58 residues absent, its potency was 3 orders of magnitude lower than that at the wildtype receptor. After truncation of 114 residues, there was no response to calcitonin. In contrast, small-molecule ligands were fully active at receptors having up to 149 amino-terminal residues absent. Those compounds finally became inactive after truncation of 153 residues. Deletion and/or alanine replacement of the region of the calcitonin receptor between residues 150 and 153 resulted in marked reduction in cAMP responses to these compounds, with some compound-specific differences observed, supporting a critical role for this region. Binding studies further supported distinct sites of action of small molecules relative to that of calcitonin. These findings focus attention on the potential importance of the juxtamembranous region of the amino terminus of the Family B calcitonin receptor for agonist drug action.Calcitonin (CT), 3 a 32-amino acid peptide secreted by the thyroid gland in response to elevations in blood calcium levels, acts on bone and kidney to maintain calcium homeostasis (1, 2). CT is widely used therapeutically for the treatment of bonerelated disorders such as osteoporosis, hypercalcemia of malignancy, and Paget disease (1, 2). This peptide must be administered parenterally, whereas small-molecule agonist ligands that can be administered orally have substantial clinical advantage, particularly for long-term treatment.CT acts on the CT receptor, a Family B G protein-coupled receptor (GPCR). Members of this family include receptors for secretin, vasoactive intestinal polypeptide, parathyroid hormone, corticotrophin-releasing factor, glucagon, glucagon-like peptide 1, CT, and CT gene-related peptide (3), with each having a long extracellular amino-terminal domain containing six conserved cysteine residues that form three intradomain disulfide bonds. This region has been shown to play a critical role in natural peptide ligand binding and receptor activation (4 -11).Development of small-molecule ligands for Family B GPCRs represents an area of great interest. For the CT receptor, several such ligands have been developed (12)(13)(14). However, the mechanisms of their actions at their receptors remai...

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“…12) In brief, cells were cultured to confluence in 96-well plates for 1-3 d. After being washed with Hanks' balanced salt solution containing 0.1% bovine serum albumin, cells were pre-incubated with the same buffer supplemented with 0.5 mM of 1-methyl-3-isobutylxanthine (MIX), 14) a phosphodiesterase inhibitor, at 37°C for 10 min, then incubated with samples at 37°C for 1 h. After removal of the supernatant, the intracellular cAMP was extracted by lysing the cells with 0.1 ml of ice-cold 0.1 N HCl; they were then neutralized with 0.5 N NaOH and measured by radioimmunoassay using a cAMP …”

Section: Reagents and Cellsmentioning

confidence: 99%

Trichorzin HA V, a Member of the Peptaibol Family, Stimulates Intracellular cAMP Formation in Cells Expressing the Calcitonin Receptor.

Katayama

Miyagawa

Kodama

et al. 2001

Biological & Pharmaceutical Bulletin

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By a cell-based screening of an in-house natural product library, trichorzin HA V, belonging to a peptaibol family, was isolated from a strain of fungus Trichoderma as a calcitonin (CT) agonist. Like CT, trichorzin HA V elevated cAMP levels in T47D cells which endogenously express the human CT receptor. It also stimulated cAMP formation in cells expressing recombinant human CT receptor, but not in those that do not express the receptor, suggesting that it selectively interacts with the CT receptor. In contrast to trichorzin HA V, alamethicin, another well-characterized peptaibol, showed no cAMP-elevating activity at all. These results suggest that, although there was little amino acid sequence similarity between trichorzin HA V and CT, the biological activity of trichorzin HA V can mimic that of CT, acting via the CT receptor.

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Discovery of a non-peptide small molecule that selectively mimics the biological actions of calcitonin

Cited by 20 publications

References 26 publications

Formulation, stability and pharmacokinetics of sugar-based salmon calcitonin-loaded nanoporous/nanoparticulate microparticles (NPMPs) for inhalation

Formulation, stability and pharmacokinetics of sugar-based salmon calcitonin-loaded nanoporous/nanoparticulate microparticles (NPMPs) for inhalation

Juxtamembranous Region of the Amino Terminus of the Family B G Protein-coupled Calcitonin Receptor Plays a Critical Role in Small-molecule Agonist Action

Trichorzin HA V, a Member of the Peptaibol Family, Stimulates Intracellular cAMP Formation in Cells Expressing the Calcitonin Receptor.

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