Bisphosphonate Compounds Research Articles

Bisphosphonates for osteoporosis treatment are associated with reduced breast cancer risk

Background:Bisphosphanates are used primarily for the prevention and treatment of osteoporosis, and are also indicated for osseous complications of malignancy. In addition to their bone resorption properties, the most commonly used nitrogen-containing bisphosphonate compounds also inhibit protein prenylation, and thus may exert anti-tumour properties.Methods:To evaluate whether the use of these drugs may be associated with cancer, specifically breast cancer, we conducted a population-based case–control study in Wisconsin from 2003 to 2006. Participants included 2936 incident invasive breast cancer cases and 2975 population controls aged <70 years. Bisphosphonate use and potential confounders were assessed by interview.Results:Using multivariable logistic regression, the odds ratio for breast cancer in current bisphosphonate users compared with non-users was 0.67 (95% confidence interval 0.51–0.89). Increasing duration of use was associated with a greater reduction in risk (P-trend=0.01). Risk reduction was observed in women who were not obese (P-interaction=0.005).Conclusion:These results are suggestive of an additional benefit of the common use of bisphosphonates, in this instance, the reduction in breast cancer risk.

Locally Delivered Bisphosphonate for Enhancement of Bone Formation and Implant Fixation

Bobyn, J. Dennis PhD1; McKenzie, Kimberly BSc2; Karabasz, Dorota BSc2; Krygier, Jan J. CET2; Tanzer, Michael MD2 Author Information

Cancer‐induced bone loss and associated pain‐related behavior is reduced by risedronate but not its phosphonocarboxylate analog NE‐10790

Prostate, breast and lung cancers readily develop bone metastases which lead to fractures, hypercalcemia and pain. Malignant growth in the bones depends on osteoclast-mediated bone resorption and in this regard bisphosphonate compounds, which have high-bone affinity and inhibit osteoclast activity, have been found to alleviate bone cancer symptoms. In this study, the bisphosphonate risedronate and its phosphonocarboxylate derivative NE-10790 was tested in a murine bone cancer pain model. Risedronate decreased bone cancer-related bone destruction and pain-related behavior and decreased the spinal expression of glial fibrillary acidic protein, whereas NE-10790 had no effect on these parameters. Furthermore, risedronate but not NE-10790 induced dose-dependent toxicity in NCTC-2472 cells in vitro. Furthermore, the direct toxic effect of risedronate on tumor cells observed in vitro opens the possibility that a direct toxic effect on tumor cells may also be present in vivo and be related to the efficacy of bisphosphonate compounds. In conclusion, these results suggest that risedronate treatment may lead to an increased life quality, in patient suffering from bone cancer, in terms of decreased osteolysis and pain, and merits further study.

Direct stability-indicating method development and validation for analysis of etidronate disodium using a mixed-mode column and charged aerosol detector

This paper describes the development and validation of a rapid, direct, and stability-indicating method for analysis of etidronate, a bisphosphonate compound without a UV chromophore. A mixed-mode column was used to separate etidronate from its impurities in an 8-min gradient method and a charged aerosol detector (CAD) was used for detection. The developed HPLC method was validated with respect to specificity, linearity, accuracy, precision, sensitivity, and stability. The method can be used for release and stability testing of etidronate and has applicability to other similar bisphosphonate compounds.

A facile one pot synthesis of bisphosphonic acids and their sodium salts from nitriles

A general and one pot synthesis for the preparation of bisphosphonic acids and their sodium salts (2a-e) from nitriles (3a-e) is described. This method involves hydrolysis of nitriles (3a-e) to the corresponding acids and subsequent bisphosphonation in a single solvent to produce bisphonates (2a-e). Preparations of some of bisphosphonates, which are presently in clinical use like risedronate (2a) sodium, ibandronate sodium (2d) are synthesized by following this new method. This method is useful for the preparation of other bisphosphonate compounds.

Spontaneous bilateral subcapital hip fracture due to multiple myeloma: an 18-month follow-up

We present a rare case of bilateral subcapital hip fracture as a first sign of multiple myeloma in a young man. Right femoral neck fracture was treated by three Asnis screws and the left one by hemiarthroplasty. In order to avoid implant failure, we used zoledronic acid, a bisphosphonate compound, instead of methacrylate cement. After an 18-month follow-up, there was no evidence of avascular necrosis, no implant failure and no periprothetic fracture.

Synthesis of Pyrophosphate-Containing Compounds that Stimulate V&amp;#947;V&amp;#948;2 T Cells: Application to Cancer Immunotherapy

Human Vgamma2Vdelta2 T cells recognize nonpeptide antigens, such as isoprenoid pyrophosphomonoester intermediates, alkylamine compounds, and bisphosphonate drugs, as well as some tumor cells. Although attempts have been made to derive novel cancer immunotherapies based on the discovery of these unconventional antigens, effective therapies remain to be developed. Here, we synthesized a series of pyrophosphate-containing compounds and examined the chemical requirements for the recognition of pyrophosphomonoester antigens by gammadelta T cells. The structural analysis clearly demonstrated that a proximal methylene moiety plays a crucial role in the stimulatory activity of the antigens. For optimal gammadelta T cell proliferation, we find that the use of human serum albumin was preferred and that pyrophosphomonoesters were superior to nitrogen-containing bisphosphonate compounds. Using these techniques, we have successfully expanded gammadelta T cells from healthy donors as well as from cancer patients using one of the most active compounds, 2-methyl-3-butenyl-1-pyrophosphate (2M3B1PP). The resulting expanded gammadelta T cells exhibited potent, cytotoxic activity against a wide variety of tumor cell lines. Even gammadelta T cells from a patient with advanced liver carcinoma efficiently responded to 2M3B1PP and exhibited strong cytotoxic activity against tumor cells. The pretreatment of tumor cells with nonpeptide antigens was essential for efficient cytotoxicity via TCR-gammadelta. The present study suggests a novel strategy for cancer immunotherapy using synthetic small pyrophosphate-containing compounds and nitrogen-containing bisphosphonates.

The structural study of metal complexes of bisphosphonate amide ester

As a class of materials, metal phosphonates have been known for almost 30 years, but in recent years the research has expanded rapidly due to the important applications of metal bisphosphonate compounds in industrial processes such as ion-exchange, catalysis and sorption [1-3]. Recently, several metal complexes of clodronate, (dichloromethylene)-bisphosphonate, Cl2MBP, which is one of the best-documented bisphosphonates used effectively in medical applications, and its symmetrical dialkyl ester derivatives, have been prepared for studying the properties of solid materials [4]. The arrangement of metal complexes of bisphosphonates is dependent on the size of the organic group connected to the phosphonate groups, and also on the number of donor atoms of the ligand and on the degree of protonation of the phosphonate group [2]. In this study, a new, selective and simple method for the preparation of amide ester derivatives of Cl2MBP, has been developed. Four metal complexes of one of these synthesized ligands, P-piperidinium-P umethyl-(dichloromethylene)bis-phosphonate have been prepared in order to study the effect of the cyclic amine group on the complexing properties of the ligand and on the dimensionalities of its metal complexes. The compounds were crystallised by the gel method, and the structures of the complex crystals were characterized by single crystal X-ray diffraction methods, elemental analyses and thermal analyses. The methyl ester of piperidine derivative of Cl2MBP ligand forms the first wheel-like metal complex of bisphosphonate; the packing of the copper complex consist of channels occupied by aqua ligands and lattice water molecules. The complex, which has a unique hexameric structure, differs from rather common heptanuclear transition metal clusters such as Cu6Na, Co6Na, Fe6Na, Zn7, Mn7 and Cu7, where the seventh metal ion occupies the central position as a cation template. The other three compounds are polymers; Mg, Zn and Cd complexes consist of one-dimensional chains with different composition. In addition, in the case of the zinc complex, the coordinated Na cations hold the chains together forming a two-dimensional layered construction. Divalent metal cations are capable of forming different compounds with the methyl ester of piperidine derivative of clodronic acid. These kinds of structures provide new information about the ability of the amino derivatives of clodronic acid to form polymeric structures and microporous materials that can be utilized in chemical applications.

Recognition and Function of Human&amp;#947;&amp;#948;T Cells: Application for Tumor Immunotherapy

Human Vγ2Vδ2T cells recognize nonpeptide antigens derived from microbial pathogens in a TCR-dependent manner, such as isopentenyl pyrophosphate-related metabolites and alkyl amines. Recent crystallographic analysis of the γ2VδTCR along with the site-directed mutagenesis studies has strongly suggested that theγ2VδTCR directly interacts with nonpeptide antigens via a positively-charged pocket formed by CDR2 and CDR3 regions. Full activation ofγ2VδT cells by nonpeptide antigens leading to proliferation and cytokine production was suggested to require cognate cellular interaction with the cells displaying the antigens via specific adhesion molecules. Synthetic nitrogen-containing bisphosphonate compounds such as pamidronate were also shown to activate the same subset ofγ2VδT cells in a TCR-dependent manner. Most notably, these compounds can efficiently sensitize a wide spectrum of human tumor cells to the lysis byγ2VδT cells. These characteristics ofγ2VδT cells confer them a unique role in cross-immunity in infection and malignancy, and may provide a novel strategy for immunotherapy of human cancer. Keywords: γ2Vδt cells, nonpeptide antigens, pamidronate, cancer immunotherapy, infection immunity

Single-Dose Pharmacokinetics and Tolerability of Alendronate 35- and 70-Milligram Tablets in Children and Adolescents with Osteogenesis Imperfecta Type I

Alendronate (ALN) is a bisphosphonate compound that can be administered orally and has potential use in pediatric osteoporotic conditions. The objective was to evaluate the pharmacokinetics and single-dose tolerability of ALN in children with osteogenesis imperfecta. ALN was administered iv and orally in a two-period, randomized crossover study, with doses separated by a 2-wk washout and follow-up carried out within 2 wk after the last ALN dose. The study was conducted at the pediatric metabolic bone research unit at the Shriners Hospital for Children, Montréal, Canada. Twenty-four children (aged 4-16 yr; eight girls) with osteogenesis imperfecta type I participated. All patients received iv ALN at a dose of 125 mug. In addition, patients weighing less than 40 kg received an oral dose of ALN 35 mg, whereas those weighing 40 kg or more received ALN 70 mg orally. Total urinary excretion and oral bioavailability of ALN, blood and urine safety parameters, and adverse events were the main outcome measures. The total urinary excretion of ALN after the iv dose was similar for both weight groups. The mean oral bioavailability (95% confidence interval) was 0.43% (0.28, 0.64%) for patients weighing less than 40 kg and 0.56% (0.36, 0.87%) for patients weighing 40 kg or more. Eighteen patients reported a total of 44 clinical adverse experiences, none of which were serious. The most common adverse experiences were mild to moderate headache (n = 7), nausea (n = 7), fever (n = 5), and abdominal pain (n = 6). Eighty percent of the adverse experiences (35 of 44) occurred within 48 h of medication administration, 91% (40 of 44) lasted less than 24 h, and 84% (37 of 44) were reported after oral dosing. Laboratory safety monitoring revealed a marginal decrease in absolute lymphocyte count and serum alkaline phosphatase after the study compared with baseline for both weight categories. The mean oral bioavailability of 35- and 70-mg ALN tablets was less than 0.6%, comparable to adult studies. Adverse experiences from single-dose ALN were minor, and the drug was generally well-tolerated.

Efficacy of oral etidronate for skeletal diseases in Japan.

Etidronate is an oral bisphosphonate compound that is known to reduce bone resorption through the inhibition of osteoclastic activity. The efficacy of etidronate for involutional (postmenopausal and senile) and glucocorticoid-induced osteoporosis, as well as that for other skeletal diseases, was reviewed in Japanese patients. Cyclical etidronate treatment (200 mg or 400 mg/day for 2 weeks about every 3 months) increases the lumbar bone mineral density (BMD) in patients with involutional osteoporosis and prevents incident vertebral fractures in patients with glucocorticoid-induced osteoporosis. The losses of the lumbar BMD in patients with liver cirrhosis and the metacarpal BMD in hemiplegic patients after stroke are prevented, and the lumbar BMD is possibly increased, preventing fragile fractures in adult patients with osteogenesis imperfecta type I. Furthermore, proximal bone resorption around the femoral stem is reduced and some complications may be prevented in patients who undergo cementless total hip arthroplasty. Oral etidronate treatment may also help to transiently relieve metastatic cancer bone pain followed by a decrease in abnormally raised bone resorption in patients with painful bone metastases from primary cancer sites, such as the lung, breast and prostate. Thus, oral etidronate treatment is suggested to be efficacious for osteoporosis, as well as other skeletal diseases associated with increased bone resorption, in Japanese patients. Randomized controlled trials needed to be conducted on a large number of patients to confirm these effects.

Effect of pamidronate treatment in children with polyostotic fibrous dysplasia of bone.

Intravenous infusions with the bisphosphonate compound pamidronate decrease bone pain and reportedly can lead to refilling of dysplastic lesions in adults with fibrous dysplasia (FD) of bone. Here we describe the effects of this treatment approach in 18 children and adolescents (age at start of therapy, 6.2-17.5 yr; eight girls) with polyostotic FD, who received pamidronate for 1.2-9.1 yr (median, 3.8 yr). Treatment cycles with pamidronate (1-1.5 mg/kg.d on 3 consecutive days) were given every 4 months. Levels of serum alkaline phosphatase and urinary collagen type I N-telopeptide were elevated at baseline and decreased continuously during the first 3 yr of therapy. There was no radiographic evidence of filling of lytic lesions or thickening of the bone cortex surrounding the lesions in any patient. Histomorphometric results in dysplastic bone tissue of patients receiving pamidronate (n = 7; time of therapy, 1.4-4.8 yr) were similar to those of patients without medical therapy (n = 9). No serious side effects were noted. In conclusion, pamidronate therapy appears to be safe in children and adolescents with polyostotic FD. However, we found no clear evidence that pamidronate has an effect on dysplastic lesions in such patients.

Superficial gyrate erythema as a cutaneous reaction to alendronate for osteoporosis

Alendronate is a bisphosphonate compound used to treat diseases of pathologic bone resorption. Cutaneous reactions to this medication are rare. We report the case of a superficial gyrate erythema that occurred as a reaction to weekly alendronate therapy used in the treatment of postmenopausal osteoporosis. (J Am Acad Dermatol 2003;48:945-6.)

Growth Inhibition of Macrophage-like and Other Cell Types by Liposome-encapsulated, Calcium-bound, and Free Bisphosphonates In Vitro

Bisphosphonates effectively inhibit osteoclastic bone resorption in diseases characterized by excessive bone loss. Liposome-encapsulated clodronate (dichloromethylene bisphosphonate) also is known to inactivate phagocytic cells in vivo, and inhibit the growth of macrophage-like Raw 264 cells in vitro. The macrophage suppressive effect of liposomal clodronate is of interest in autoimmune diseases, like rheumatoid arthritis, in which phagocytic cells are involved in inflammatory processes. Earlier in vivo studies suggested that liposomal clodronate is a far more potent inactivator of macrophages than liposomal forms of two other bisphosphonate compounds, pamidronate (3-amino-1-hydroxypropylidene bisphosphonate), and etidronate (1-hydroxyethylidene-1, 1-bisphosphonate). We examined the growth inhibitory properties of these three bisphosphonates with macrophage-like RAW 264 cells and with other types of cells in vitro. All three bisphosphonates encapsulated in liposomes effectively inhibited the growth of RAW 264 and CV1-P cells, while free drugs were 20–1000 times less potent growth inhibitors. Also, high extracellular calcium concentrations enhanced the potency of bisphosphonates for RAW 264 cells, indicating that, in addition to liposomes, the uptake of bisphosphonates by macrophages, is mediated also by calcium. In all formulations, pamidronate was the most potent compound for the cells, with the exception CV1-P cells, for which liposomal clodronate was the most potent. The effects of liposomal drugs were selective for highly endocytotic cells. The results suggest that liposome-encapsulated bisphosphonates could provide a specific tool to affect the function of macrophages and all three of these bisphosphonates are potentially effective as macrophage suppressors in autoimmune diseases.

Novel antiangiogenic effects of the bisphosphonate compound zoledronic acid.

Bisphosphonate drugs inhibit osteoclastic bone resorption and are widely used to treat skeletal complications in patients with tumor-induced osteolysis. We now show that zoledronic acid, a new generation bisphosphonate with a heterocyclic imidazole substituent, is also a potent inhibitor of angiogenesis. In vitro, zoledronic acid inhibits proliferation of human endothelial cells stimulated with fetal calf serum, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (IC(50) values 4.1, 4.2, and 6.9 microM, respectively), and modulates endothelial cell adhesion and migration. In cultured aortic rings and in the chicken egg chorioallantoic membrane assay, zoledronic acid reduces vessel sprouting. When administered systemically to mice, zoledronic acid potently inhibits the angiogenesis induced by subcutaneous implants impregnated with bFGF [ED(50), 3 microg/kg (7.5 nmol/kg) s.c.]. These findings indicate that zoledronic acid has marked antiangiogenic properties that could augment its efficacy in the treatment of malignant bone disease and extend its potential clinical use to other diseases with an angiogenic component.

New bone-seeking agent: Animal study of Tc-99m-incadronate

Disodium cycloheptylaminomethylenediphosphonate monohydrate (incadronate disodium) is a third-generation bisphosphonate compound which potently inhibits bone resorption, and a highly effective drug in the treatment of metastatic bone disease. We first labeled incadronate disodium with 99mTc, and examined its biodistribution and bone uptake after intravenous injection in rats to assess its potential for clinical use as a bone-seeking agent for judgment of the therapeutic effect of incadronate on bone metastases. Bone scan with 99mTc-labeled incadronate (99mTc-incadronate) may yield important information prior to the use of incadronate for treatment of bone metastases. Synthesis of 99mTc-incadronate was carried out by reduction of 99mTc-pertechnetate in the presence of SnCl2 and N2 gas. Normal rats were injected with 18.5 MBq (0.5 mCi) 99mTc-incadronate in a volume of 0.1 ml intravenously and then sacrificed at 15 min, 30 min, 1 h or 2 h (six rats at each time point) after injection. Samples of muscle, stomach, small intestine, kidney, liver and bone (femur) were taken and weighed. In addition, a 1-ml sample of blood was drawn from the heart, and urine was taken from the urinary bladder immediately after sacrifice. Samples were measured for radioactivity and expressed as percent uptake of injected dose per gram or per milliliter (% ID/g or ml). Bone-to-blood and bone-to-muscle uptake ratios were determined from the % ID/g or ml values for these organs. The greatest accumulation of 99mTc-incadronate was found in bone. Radioactivity in bone was as high as 3.22 +/- 0.68% ID/g at 2 hours after injection. Scintigraphic images of 99mTc-incadronate in normal rats revealed highly selective skeletal uptake. 99mTc-incadronate exhibited high uptake in bone, and relatively low uptake in soft tissue, suggesting that it may be useful as a bone-seeking agent for judgment of the therapeutic effect of incadronate on bone metastases, by determining the degree of its accumulation in metastatic bone lesions.

Electrospray ionization mass spectrometry and tandem mass spectrometry of clodronate and related bisphosphonate and phosphonate compounds

The bisphosphonate family with a P-C-P structure is a broad class of drugs, widely investigated as potential inhibitors in bone diseases and calcium metabolic disorders. In this study, the mass spectrometric (MS) behavior and fragmentation of clodronate and related bisphosphonate and phosphonate compounds was studied by using negative ion electrospray ionization (ESI) with triple quadrupole and ion trap instruments. The effect of pH on the degree of deprotonation of the polyprotic bisphosphonic and phosphonic acids in negative ion ESI-MS was investigated, and the degree of deprotonation in the ESI mass spectra and the dissociation in the liquid phase were compared. The results provide evidence that the measured ESI mass spectra do not correlate with the chemistry in the liquid phase owing to the decrease in the pH of the solvent droplets during the ion evaporation process and the charge state neutralization in the gas phase. Ion trap MS(n) provided useful information on the fragmentation study of clodronate and related bisphosphonate and phosphonate compounds, in which interesting fragmentation pathways including the direct elimination of carbon monoxide from deprotonated bisphosphonates and formation of a P-P bond were observed. Reactions between the product ions with a -PO(2) group and residual water in the ion trap or in the high-pressure region of the triple quadrupole instrument formed other unexpected fragmentation paths for all the bisphosphonates studied.

Pharmacological and clinical properties of alendronate sodium hydrate

Alendronate (alendronate sodium hydrate; Bonalon Tablet, 5 mg) is a nitrogen-containing bisphosphonate, which combines with the bone surface and reduces osteoclast-mediated bone resorption. It is a third-generation bisphosphonate compound, specifically distributed on the surface of bone resorption and taken into osteoclasts. Under the closed circumstances which is formed with osteoclast and the bone surface, alendronate becomes detached from the bone surface and taken into osteoclast since acid released from osteoclast leads to pH decrease (acidified). The uptaken alendronate blocks the pathway of mevalonic acid synthesis, which is cholesteric synthesis, inhibits the prenylation of GTP binding protein, and decreases the osteoclast's function by influencing the cytoskeleton. This restraint of alendronate in bone resorption against osteoclasts is reversible, showing no cytotoxicity at more than hundredfold concentration level at which action occurs. Alendronate is an agent for the treatment of osteoporosis that has established safety with regards to bone quality since it neither inhibits bone calcification nor influences fracture healing in chronic administration. The most serious morbidity in osteoporosis is developing fractures. The efficacy of alendronate on restraining fracture, as well as on increase in BMD, is evidenced in Japan. Recently, in addition to senile or postmenopausal osteoporosis, drug-induced osteoporosis, such as steroid-induced osteoporosis, has attracted attention. In this regard, alendronate has been found to be an effective agent for the treatment of osteoporosis overseas, being approved in over 90 countries and used by more than 4.5 million patients. This review will give an outline of alendronate, the preparation to have introduced a concept of Evidence Based Medicine earlier, from pharmacodynamic action to clinical efficacy.

Ion-exchange liquid chromatographic analysis of bisphosphonates by on-line post-column photochemical reaction and spectrophotometric detection

A simple ion-exchange high-performance liquid chromatographic method was developed and employed for the analysis of bisphosphonate compounds in dosage formulations using on-line post-column photochemical reactions. The method used molybdate as the post-column reagent to react with the photolyzed bisphosphonate to form phosphomolybdate for enhanced spectrophotometric detection. A bisphosphonate compound, 2-thioethane-1,1-bisphophonic acid, was selected to evaluate the separation using both isocratic and gradient elution methods, along with the effects of other experimental parameters including mobile phase composition, flow-rate and post-column reagent concentration. The gradient elution method showed improved resolution and detection sensitivity compared to the isocratic elution method. The optimized gradient method was simple, reproducible, and specific to bisphosphonate compounds. It was successfully employed for the stability study of the bisphosphonate compound in pharmaceutical dosage formulations.

Etridronate therapy in the treatment and prevention of osteoporosis.

Etidronate disodium is an oral bisphosphonate compound known to reduce bone resorption through the inhibition of osteoclastic activity. This article is a review of its efficacy and safety in the treatment and prevention of postmenopausal and corticosteroid-induced osteoporosis. In general, studies of cyclical etidronate therapy (400 mg daily for 2 wk every 3 mo) have found a significant improvement in bone density. These studies have not been powered to study fracture incidence, but a reduced fracture rate has been found in some of the studies reviewed. Studies examining cyclical etidronate in the prevention of osteoporosis indicate it prevents early menopausal bone loss and is free of significant side effects. In both prevention of corticosteroid-induced osteoporosis and treatment of patients who have been on long-term corticosteroid therapy, cyclical etidronate appears to increase bone density and prevent further loss of bone. In summary, a review of available literature pertaining to the use of etidronate in prevention and treatment of primary and secondary osteoporosis has been presented. This review suggests etidronate, used as a cyclical therapy, is a safe and effective therapy. The weight of evidence suggests it is capable of reducing fracture risk in patients with osteoporosis. Increases in bone density at the spine and hip are not as pronounced as with some other bisphosphonates, particularly alendronate, but no direct clinical comparison trials of significant size or duration have been undertaken.