KR20050110814A - Compositions and methods for inhibiting bone resorption - Google Patents

Abstract

Compositions and methods are described for preventing, inhibiting, reducing and treating conditions and diseases associated with abnormal bone absorption in mammals, such as osteoporosis. Embodiments of the compositions of the present invention comprise a pharmaceutically effective amount of alendronate and vitamin D ₃ suitable for one week administration. The compositions and methods of the present invention provide vitamin D nutrition during bisphosphonate treatment to facilitate normal bone formation and mineralization, while minimizing the induction of potential for complications associated with vitamin D deficiency (eg, hypocalcemia and osteomalacia). Also described are methods of making the compositions of the present invention, methods of measuring stability and degradability of the compositions, and methods of measuring plasma levels of vitamin D.

Description

Compositions and methods for inhibiting bone resorption

The present invention relates to a composition comprising a bisphosphonate compound and a vitamin D compound. The invention also relates to methods of using the compositions to treat, reduce, inhibit or prevent abnormal bone absorption, for example in mammals. The invention also relates to a process for the preparation of bisphosphonates and vitamin D compositions.

Various diseases in humans and mammals include or are associated with abnormal bone absorption. The most common of these diseases is osteoporosis, which is a systemic skeletal disease characterized by low bone mass and a decrease in the microstructure of bone tissue, and thus a subsequent increase in bone fragility and susceptibility to fracture. Osteoporosis is a pandemic pandemic, with a significant increase in incidence in line with global life expectancy.

The major cell types involved in bone resorption are multinuclear cells called osteoclasts. Bisphosphonates are well known as selective inhibitors of osteoclast bone uptake. Bisphosphonates bind to hydroxyapatite in bone and are believed to inhibit bone resorption of osteoclasts through their intracellular action. H. Fleisch, Bisphosphonates In Bone Disease, From The Laboratory To The Patient, 4th Edition, Academic Press (2000)]. It has also been reported that bisphosphonates bind to bone and then are released into the absorption space during absorption. Thereafter, they are taken up by osteoclasts and then inhibit the enzyme farnesyl diphosphate synthase. This intracellular action also prevents isoprenylation (parnesylation and geranylgeranylation) of GTPases, signaling proteins that bind to membrane vesicles. Geranylgeranylated small groups of GTPases include those involved in the formation of mesenteric margins-organelles of active bone uptake [Ref. A.A. Reszka and G.A. Rodan, "Bisphosphonate Mechanism of Action," Curr Rheumatol Rep. 5 (1): 65-74 (February, 2003)].

It can be seen that bisphosphonates are useful for preventing bone loss associated with numerous conditions. For example, bisphosphonates prevent, but are not limited to, bone loss, osteoporosis, osteopenia, metastatic bone disease, multiple myeloma, periodontal disease, tooth loss, parathyroidism, rheumatoid arthritis, Paget's disease, It is known to be useful in the treatment of diseases such as osteonecrosis, osteoarthritis, peritonitis bone loss or osteolysis, and hypercalcemia of malignant tumors. All these conditions are characterized by bone loss resulting from an imbalance between bone uptake—ie destruction—and bone formation.

Alendronate sodium is one of the most potent bisphosphonates on the market and does not affect bone mineralization at a dose that inhibits bone absorption as much as possible. In addition, the increase in bone minerals observed upon administration of alendronate has been found to be positively associated with decreased spinal and non-vertebral (including hip) fractures, reduced spinal cord malformations, and kidney maintenance. This means that when administered for a substantial period of time, alendronate reduces bone turnover, which acts positively to produce fortified bone. Alendronate sodium has been approved in more than 90 countries for the treatment of osteoporosis in postmenopausal women. Alendronate sodium has also been approved for the treatment of osteoporosis, glucocorticoid-induced osteoporosis in men and Paget's disease of bone. Evidence shows that other bisphosphonates, such as ibandronate, minodronate, pamidronate, risedronate, tiludronate, and zoledrnate, have high efficacy as inhibitors of osteoclast bone resorption, It has a characteristic.

Despite their therapeutic benefits, bisphosphonates are poorly absorbed from the gastrointestinal tract (levels of about 1%) [B.J. Gertz et al, Clinical Pharmacology of Alendronate Sodium, Osteoporosis Int., Suppl. 3: S13-16 (1993) and B.J. Gertz et al., Studies of the oral bioavailability of alendronate, Clinical Pharmacology & Therapeutics, vol. 58, number 3, pp. 288-298 (September 1995)]. It will be appreciated that foods (including beverages, such as mineral water, and even some excipients used to formulate administration vehicles), as well as many other substances that can be ingested, can adversely affect bisphosphonate absorption. Intravenous administration was used to bring the entire dose to circulation. Intravenous administration, however, is costly and inconvenient, especially if the subject is to be repeatedly given intravenous infusions lasting several hours. Unlike oral administration, intravenous administration of bisphosphonates is associated with acute kidney injury when administered too quickly.

Instead of intravenous administration, more doses may be administered to compensate for the low bioavailability from the gastrointestinal tract, if oral administration of bisphosphonates is desired. To compensate for this low bioavailability, it is generally recommended that the subject absorb bisphosphonates in the empty stomach and then fast for at least 30 minutes. However, many subjects found this fasting inconvenient for one day.

Bisphosphonate therapy has been associated with hypocalcemia. During treatment with bisphosphonates, initial inhibition of bone resorption can lead to a decrease in serum calcium occurring within hours, days or weeks of initiation of treatment. Serum calcium reduction can last for weeks to months after initiation of treatment and can be significant in vitamin D-overlapped subjects. Hypocalcemia response to bisphosphonate therapy may in some cases be sufficiently sufficient to be symptomatic and valid clinical intervention, especially in patients with parathyroid hyperplasia [Vasikaran, SD, Ed., Bisphosphonate: An Overview with Special Reference to Alendronate, Ann. Clin. Biochem. (2001) 38: 608-623. Since there is no substantial body storage of calcium outside bone, the calcium needed for new bone formed after starting treatment with alendronate must be absorbed from dietary-food or calcium supplements. Vitamin D is necessary for normal calcium absorption. Therefore, adequate vitamin D and calcium uptake are desirable for subjects using bisphosphonates. Appropriate vitamin D levels are becoming increasingly important when calcium demand is elevated due to the net inflow of calcium into the bone resulting from bisphosphonate therapy during effective osteoporosis treatment. As a result, adequate vitamin D and calcium intake are desirable for subjects using bisphosphonates.

Vitamin D compounds contain a group of fat soluble secosteroids found in very few natural foods, which are photosynthesized in the skin of vertebrates by the action of UV radiation from the sun. Vitamin D may exist in several forms, but the most physiologically relevant forms are vitamin D ₃ (cholecalciferol) and vitamin D ₂ (ergocalciferol). Vitamin D ₂ is formed when yeast and plant sterols, ergosterols are exposed to UV radiation, while vitamin D ₃ is produced from 7-dehydrocholesterol and synthesized in the skin. Metabolic pathways for vitamin D ₃ and vitamin D ₂ are similar, their biological potency in humans is similar, and their main function is the maintenance of serum calcium and phosphorus concentrations within the normal range. Vitamin D ₃ is an essential precursor of the hormone calcitriol (also called 1,25-dihydroxycholecalciferol or 1,25-dihydroxyvitamin D ₃ ), the main action of which is the small intestine's ability to absorb calcium To maintain phosphate from the diet. The hormone-type metabolite of ergocalciferol is 1,25-dihydroxyergocalciferol (1,25-dihydroxyvitamin D ₂ ). When dietary calcium absorption is insufficient to meet body needs, parathyroid hormone (PTH), along with calcitriol, a hormone metabolite of vitamin D ₃ , activates the bone marrow mononuclear stem cells to mature osteoclasts. These osteoclasts themselves are stimulated by various cytokines and other factors to increase the mobilization of calcium storage from bone.

The natural-types of cholecalciferol and ergocalciferol are biologically inert precursors of the hydroxylated biologically active metabolites of vitamin D. Because vitamin D is lipid soluble, it can be stored in the body's adipose tissue or metabolized to 25-hydroxyvitamin D, the primary storage metabolite, and stored in other organs. 25-hydroxyvitamin D is transported into the plasma and metabolized by the body as needed. In particular, as shown as an example of cholecalciferol in FIG. 1, 7-dehydrocholesterol in the skin is followed by pre-vitamin D ₃ (isomer of vitamin D ₃ , not shown in FIG. 1), followed by vitamin Is converted to D ₃ (cholecalciferol). The cholecalciferol is then metabolized in the liver to form 25-hydroxycholecalciferol, also known as 25-hydroxy vitamin D ₃ , which is also a hormone form known as calcitriol in the kidney 1,25- Metabolized to dihydroxycholecalciferol. Although not shown in FIG. 1, other metabolites of vitamin D (D ₂ or D ₃ ) include 1α-hydroxy vitamin D, 24,25-dihydroxy vitamin D and 1α, 24,25-trihydroxy vitamin D. Include. Only calcitriol is fully biologically active, and cholecalciferol and the metabolites identified above show little biological activity.

The main biological action of vitamin D (both D ₂ or D ₃ ) is to help maintain calcium homeostasis by increasing the efficacy of the gut to absorb dietary calcium. This helps to ensure that the amount of calcium absorbed is suitable for maintaining blood calcium in the normal range and for maintaining skeletal mineralization. Proper vitamin D intake facilitates intestinal absorption of calcium and plays an important role in regulating calcium metabolism and mineralization of the skeleton.

Vitamin D dysfunction and deficiency is recognized as a cause of metabolic bone disease in adults. Vitamin D dysfunction is characterized by disorders of calcium and phosphate uptake, but no normal bone mineralization disorder, and is usually associated with serum 25-hydroxy vitamin D levels of about 9 or less to about 30 ng / ml. Vitamin D deficiency is characterized by severely impaired calcium absorption, secondary hyperparathyroidism, hypophosphatemia, low or normal blood calcium and impaired bone mineralization. Serum 25-hydroxy vitamin D levels are usually about <9 ng / ml. Vitamin D dysfunction and deficiency lead to an increase in parathyroid hormone (PTH), which also results in increased osteoclast activity, phosphate loss in urine and calcium mobilization from bone. It can also exacerbate osteoporosis, especially in older adults, as impaired bone mineralization causes an independent and additional decrease in bone strength. Sustained vitamin D dysfunction is considered to be an important cause of gradual bone loss. Depending on the extent of vitamin D and calcium deficiency, the histological composition may be osteomalacia, osteoporosis or one of the two.

The prevalence of vitamin D dysfunction and deficiency causes the need for additional vitamin D intake in groups of patients susceptible to or suffering from conditions such as osteoporosis or osteopenia, and subjects performing bisphosphonate treatment of these conditions. In subjects performing bisphosphonate treatment, and in particular those with improper dietary calcium intake or inadequate calcium uptake, adequate vitamin D is necessary to facilitate bone formation and mineralization while minimizing the potential or occurrence of vitamin D dysfunction. . Several forms of increasing vitamin D uptake are often used in clinical trials of bone absorbing compounds and are presented in product labeling and product package circulation. However, for example, approximately 30% of osteoporosis patients in the United States show some degree of vitamin D dysfunction and an increase in prevalence with age.

Typically, subjects who are bisphosphonate administered orally and need vitamin D are advised to receive two separate products at two different times. Vitamin D formulations are most commonly administered daily, while bisphosphonates can be administered daily, weekly, monthly, or at longer intervals. As a result, even though many patients being treated for osteoporosis or osteopenia have been advised to do so, they do not take vitamin D. Typically, vitamin D has poor bisphosphonate absorption, and most bisphosphonate oral feeding regimens require a 30 minute time interval between digestion of bisphosphonates and other substances, including but not limited to vitamin D. The simple fact is that you cannot take it simultaneously with bisphosphonates. As a result, few patients follow a dosage regimen that requires separate administration of vitamin D compounds at regular time intervals before or after bisphosphonate administration (some bisphosphonate compounds require administration prior to digestion of the food, thus vitamin D is bisphosphonate). Should be administered at regular time intervals after administration). Patients may take vitamin D before or after receiving their bisphosphonates, but there is evidence that many patients do not. A marketing study in 1998 found that 75-85% of physicians who prescribe alendronate also recommend vitamin D supplementation, but only 57% of patients with osteoporosis actually follow.

Vitamin D can also be administered in the form of multi-vitamins in the United States, but, for example, many commercially available oral vitamin D formulations are not marketed in dosage units necessary to be administered less frequently than a day. If the patient himself is taking vitamin D at the same time as their bisphosphonate administration, the form of vitamin D administered will prevent bisphosphonate absorption because many vitamin D compounds formulated for osteoporosis patients contain calcium which reduces the absorption of bisphosphonates. It can also interfere and reduce.

Patent literature includes patents and published patent applications that describe vitamin D ₃ or vitamin D ₂ or their metabolites or homologues, together with bisphosphonates. See, for example, US Pat. Nos. 4,230,700, 4,330,537, and Pat. 4,812,304; EP 0 381 296 and EP 0 162 510; International Patent Publications WO 90/01312, WO 92/21355, WO 01/28564, WO 01/97788 and WO 03/086415; European Patent Publication EP 1 051 976; Japanese Patent Publication Nos. 7-330613 and 11-60489; US Patent Application Nos. US 2003/0139378 A1 and US 2003/0225039 A1]. However, these patents and publications are compositions, products or formulations comprising bisphosphonate compounds and vitamin D compounds that are useful for continuous oral administration at intervals such as once a week, such as less frequently than one day and more often than six months or more. (And, most particularly, tablets), may not be described or enabled. These patents and publications also describe methods for treating, inhibiting, reducing or preventing other conditions associated with abnormal bone absorption and osteoporosis by administering such bisphosphonate / vitamin D compositions less frequently than daily, at more frequent intervals than 6 months, or , Can not be enabled.

As a result, there is a need for a combination product comprising a bisphosphonate compound and a vitamin D compound, including enhancing the overall efficacy of the bisphosphonate therapy by helping to ensure adequate vitamin D uptake to facilitate calcium absorption. In addition, there is a need for vitamin D and bisphosphonate products that facilitate normal bone formation and mineralization while reducing or minimizing the potential for or occurrence of complications associated with vitamin D dysfunction (eg, hypocalcemia and osteomalacia). . There is a need for bisphosphonates and vitamin D products that provide a certain amount of vitamin D nutrition that facilitates normal bone formation and mineralization in subjects performing bisphosphonate treatment. There is also a need for a bisphosphonate and vitamin D combination for oral administration on a continuous dosing schedule that is less frequent than daily administration and more frequent than six month or longer intervals. There is a need for a single product comprising vitamin D and bisphosphonates suitable for weekly administration that increases the ease of vitamin D intake and increases patients following vitamin D nutrition presented during bisphosphonate treatment. Moreover, there is a need for a method of making and administering such vitamin D / bisphosphonate compositions.

The present invention provides a pharmaceutical composition comprising a bisphosphonate compound and a vitamin D compound. Embodiments of the invention include, for example, pharmaceuticals comprising bisphosphonate compounds, or pharmaceutically acceptable salts, derivatives or hydrates of bisphosphonates, or mixtures thereof and vitamin D compounds (e.g., pharmaceutical grade vitamin D compounds). Composition. In an embodiment, the vitamin D compound comprises cholecalciferol. In an embodiment, the bisphosphonate is, for example, an alendronate, a pharmaceutically acceptable salt thereof (e.g., a sodium, potassium, calcium, magnesium or ammonium salt or a hydrate of any one of these salts) [e.g. alendronate monosodium, alendronate yl Sodium monohydrate or alendronate monosodium trihydrate]. In one embodiment of the invention, the pharmaceutical composition comprises cholecalciferol and alendronate monosodium trihydrate (see FIG. 2). The compositions of the present invention may be in the form of compressed, coated or uncoated tablets, capsules, elixirs, emulsions, or other acceptable formulations.

In an embodiment of the composition of the present invention, the bisphosphonates (or pharmaceutically effective salts, derivatives or hydrates thereof, or mixtures thereof) are present in a pharmaceutically effective amount, eg, from about 0.05 to about 1120 mg. In the same or other embodiments of the compositions of the present invention, the vitamin D compound is present in a pharmaceutically effective amount, for example, from about 100 to about 60,000 IU of vitamin D compound (vitamin D of 40 IU has a mass of approximately 1 μg). do. In another embodiment, the present invention provides about 5 to about 560 mg of bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates thereof, or mixtures thereof, based on about 100 to 36,000 IU of vitamin D compounds and bisphosphonic acid activity. It relates to a pharmaceutical composition comprising. In another embodiment, the present invention provides about 5 to about 280 mg of bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates thereof, or mixtures thereof, based on about 100 to 28,000 IU of vitamin D compounds and bisphosphonic acid activity. It relates to a pharmaceutical composition comprising. In another embodiment, the invention provides about 5 to about 280 mg of bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates thereof, or mixtures thereof, based on about 100 to 8,400 IU of vitamin D compound and bisphosphonic acid activity It relates to a pharmaceutical composition comprising a. In another embodiment, the present invention provides about 5 to about 280 mg of bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates thereof, or mixtures thereof, based on about 100 to 5600 IU of vitamin D compounds and bisphosphonic acid activity. It relates to a pharmaceutical composition comprising. In another embodiment, the present invention provides about 5 to about 280 mg of bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates thereof, or mixtures thereof, based on about 100 to 4,200 IU of vitamin D compound and bisphosphonic acid activity. It relates to a pharmaceutical composition comprising. In another aspect, one aspect of the invention comprises about 70 mg of a bisphosphonate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof, based on about 2,800 IU of vitamin D compound and bisphosphonic acid activity criteria It relates to a pharmaceutical composition. One aspect of the invention is a pharmaceutical composition comprising about 70 mg of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof, on the basis of about 2,800 IU of cholecalciferol and alendronic acid activity It is about. In another embodiment, cholecalciferol is of pharmaceutical grade.

One example of a composition of the invention is cholecalciferol granules containing alendronate sodium, cholecalciferol or an appropriate amount of cholecalciferol, and additional excipients (e.g. suitable fillers, diluents, binders, lubricants, glidants) Or a disintegrant). Other examples of compositions of the present invention include cholecalciferol granules, lactose, anhydrous lactose, microcrystalline cellulose, colloidal silicon dioxide, croscarmell, containing allendronate sodium, cholecalciferol or an appropriate amount of cholecalciferol There are tablets containing rose sodium and magnesium stearate. The compositions of the present invention are stored in an amount of 1% by weight of each isomer of cholecalciferol, for example, after storage for 24 months at a relative humidity of less than about 30 ° C. and less than about 30% to meet various purity and stability criteria Formulated to contain less than (total cholecalciferol). The compositions of the present invention may also be formulated to contain cholecalciferol with less than about 5% of total degradation after storage for 24 months at less than about 30 ° C. and less than about 30% relative humidity. For storage, the effect that ambient humidity can have on the purity and stability of the composition can be removed using a suitable package, such as an aluminum foil foam pack containing a desiccant or HDPE bottles.

In addition, the present invention includes methods for preparing the compositions and formulations described herein, as well as products made according to the methods. Embodiments of this method include preparing a powder mixture comprising bisphosphonates (e.g. alendronate), densifying the powder mixture to form a mixture, milling, mixing the mixture with the vitamin D compound to form a final granule mixture, and then Compacting the granule mixture, for example, to form a tablet. In a further embodiment, the final granule mixture can be lubricated before compacting. In one embodiment, the powder mixture comprises alendronate, colloidal silicon dioxide, lactose anhydride, microcrystalline cellulose, croscarmellose sodium and magnesium stearate. In another embodiment, the roller densifying the powder mixture forms a densified ribbon, which can be ground, mixed with cholecalciferol granules, smoothed, and pressed into a solid formulation. Advantages of the preparation methods of the present invention include increased stability of vitamin D compounds in bisphosphonate / vitamin D compositions, and increased uniformity of the particle size of the individual compounds used in the compositions of the present invention.

The alendronate powder mixture may also be premixed with one or more excipients first, followed by mixing with the remaining excipients, and then compacted with a roller.

Other methods of making bisphosphonate granules (such as alendronate granules) include, but are not limited to, wet granulation methods, as well as slugging. When bisphosphonate granules are prepared using sludge, the bisphosphonate (e.g. alendronate) powder mixture can be pressed into a non-ribbon green compact and then ground into granules which are then mixed with the vitamin D compound to mix the granule mixture Can be formed and then compressed into a solid formulation, such as a tablet. Alternatively, bisphosphonates and all excipients may be wet granulated with a granulation liquid (eg water), then dried and ground to mix with the vitamin D compound and processed into a final formulation (eg tablet).

In addition to the above-mentioned methods, direct mixing methods can also be used in which bisphosphonates, all excipients and vitamin D compounds are mixed together and then compressed into tablets or encapsulated into capsules or other solid dosage forms. Further techniques of possible methods of making bisphosphonate granules are described in US Pat. Nos. 5,358,941 and 5,882,656 and PCT Publication WO 95/29679.

A bisphosphonate / vitamin D composition can be prepared as described above, followed by a drying step to reduce the moisture level of the composition. In addition, the composition can be packaged with a desiccant to reduce the moisture level.

The invention also includes methods for preventing, reducing, inhibiting or treating metabolic bone diseases. Metabolic bone diseases include, but are not limited to, osteoporosis, postmenopausal osteoporosis, steroid-induced osteoporosis, male osteoporosis, other disease-induced osteoporosis, idiopathic osteoporosis and glucocorticoid-induced osteoporosis. The invention also includes methods for preventing, reducing, inhibiting or treating osteoporosis, conditions associated with osteoporosis and other diseases and conditions associated with abnormal bone absorption. Such other diseases and conditions may further include, for example, metabolic bone disease, hypercalcemia of malignant tumors, peritonitis osteolysis, inflammatory arthritis, and other diseases and conditions herein as identified in humans or other mammals. Alternatively, the present invention relates to a method of inducing a disease that modifies the effect on the arthritis state of a mammal, comprising administering to the mammal a therapeutically effective amount of a vitamin D / bisphosphonate composition. The present invention also induces a disease that modifies the effect on zinc bone bone necrosis in a mammal, comprising administering to the mammal a therapeutically effective amount of a vitamin D / bisphosphonate composition, and prevents bone formation or progression. And to prevent joint destruction. The invention also includes a method of reducing the risk of bone fracture in a mammal, comprising administering a unit dose of the vitamin D / bisphosphonate composition.

Aspects of this method include administering a composition of the invention to a mammal, including a human. Such compositions can be administered weekly, biweekly, monthly, bimonthly and bimonthly. In this method, vitamin D is provided by the compositions of the present invention during bisphosphonate treatment while reducing the incidence or potential for complications associated with vitamin D dysfunction. Thus, the compositions and methods of the present invention may be useful in mammals that have been found to have or are prone to vitamin D dysfunction or deficiency or that require an appropriate amount of vitamin D. In one embodiment, weekly administration to treat osteoporosis or other diseases or conditions associated with abnormal bone resorption and to minimize the risk or complications from vitamin D dysfunction is a continuous schedule until the desired therapeutic effect is achieved. Is maintained. One aspect of the method of the invention is a tablet comprising about 2,800 IU cholecalciferol and about 70 mg of alendronate or a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof, in a mammal suffering from osteoporosis. Administering once a week. In one embodiment, the therapeutic effect of a weekly administration of a vitamin D compound in a composition of the present invention is substantially the therapeutic effect of a daily dose of vitamin D, for example 400 IU, 600 IU or 800 IU per day. Similar to

The present invention also includes methods of measuring cholecalciferol (eg, stability) in pharmaceutical compositions comprising cholecalciferol and bisphosphonates. An aspect of this method involves extracting cholecalciferol from such a composition into a first solution to form a second solution, separating the sample containing cholecalciferol from the second solution, eg, reverse phase high performance liquid chromatography. Detecting the amount of cholecalciferol in the sample using photography. Embodiments of this method provide increased measurement sensitivity and to distinguish between cholecalciferol and pre-cholecalciferol, or between isomers of cholecalciferol and pre-cholecalciferol, or It can be usefully used with the compositions of the present invention to detect ferrol and pre-cholecalciferol ester adducts.

The present invention also includes a method for the determination of cholecalciferol in plasma after administration of the bisphosphonate / cholecalciferol composition of the present invention. Embodiments of this method include administering to a mammal a composition comprising alendronate and cholecalciferol, obtaining a plasma sample from the mammal, and then extracting the cholecalciferol from the plasma sample to form a first solution, 1 solution of cholecalciferol and dienophil react to form one or more die-alder addition products of cholecalciferol, and the high-performance liquid chromatography (HPLC) separation method of die-alder addition product of cholecalciferol And separating the amount of cholecalciferol in the sample using mass spectroscopy. Aspects of this method provide increased measurement sensitivity and can be usefully used with the compositions of the invention, for example, to measure the pharmacokinetic effects of administration of the compositions of the invention.

The present invention also provides a total urine, an area under the serum-concentration vs. time curve (AUC), a steady state maximum plasma concentration of a tablet comprising, for example, about 70 mg of alendronate and about 2800 IU of cholecalciferol. , including by C _max), time of the C _max (T _max) and the plasma concentration intermediate the apparent half-lives (t _{l / 2)} that are affected, a method of measuring the pharmacodynamic effects over time of administering a composition of the invention To provide.

The present invention may include, consist of, or consist essentially of the essentials and components, steps, and methods described or claimed herein, as well as essentials. Further features, advantages and aspects of the present invention, features and various advantages thereof will become more apparent from the following detailed description and the practice of the present invention.

As used herein, the term “abnormal bone resorption” may mean a degree of bone resorption that exceeds the degree of bone formation locally or throughout the skeleton, or may be related to bone formation with an abnormal structure.

"Arthritis condition" or "arthritis conditions" refers to a disease in which a part thereof is inflammatory or that has a lesion defined by a particular inflammatory state of a joint or joint, most notably a disease of rheumatoid arthritis. Academic Press Dictionary of Science Technology; Academic Press; 1st edition, January 15, 1992]. Arthritis conditions can be caused by inflammation, tumors or infections. The compositions of the present invention may also be used alone or in combination, amyloidosis, ankylosing spondylitis, bacterial arthritis, basic calcium phosphate crystal deposition disease, Behcet's disease, bursitis and tendonitis, CPPD deposition disease, calcifying tendinitis, carpal tunnel syndrome tunnel syndrome, Ehlers-Danlos syndrome, enteropathic arthritis, Felty's syndrome, myalgia, gout, fungal arthritis, hemochromatosis, hemoptymic disease, hypertrophic osteoarthritis, infectious arthritis, Inflammatory bowel disease, juvenile arthritis, juvenile rheumatoid arthritis, lupus erythematosus, Lyme disease, marfan syndrome, mixed joint tissue disease, multiple myocardial histiocytosis, myopathy, myositis, osteoarthritis, bone necrosis, osteonecrosis chondrotrophy Multiple arthritis, rheumatoid polymyalgia, psoriatic arthritis, Raynaud's phenomenon, reflex sympathetic dystrophy syndrome, Radar syndrome Arthritis such as Reiter's syndrome, recurrent polychondritis, rheumatoid arthritis, rheumatic fever, sarcoidosis, septic arthritis, scleroderma, Sjogren's syndrome, vertebral dysplasia, systemic lupus erythematosus and viral arthritis It is useful for treating or preventing arthritis conditions or symptoms / diseases comprising. In contrast to rheumatoid arthritis, osteoarthritis is a connective tissue disease, with pathology induced from epileptic seizure-induced articular cartilage degeneration, zinc bone bone remodeling and limited synovitis inflammatory response. The net result of these activities is secondary joint deformation for erosion of articular cartilage, periarticular cartilage / osteobacterialness, zinc bone necrosis and cyst formation [Oettmeier, R., and K. Abendroth, 1989] , "Osteoarthritis and bone: osteologic types of osteoarthritis of the hip," Skeletal Radiol. 18: 165-74; Cutolo M, Seriolo B, Villaggio B, Pizzorni C, Craviotto C, Sulli A. Ann. N.Y. Acad. Sci. 2002 Jun; 966: 131-42; Cutolo, M. Rheum Dis Clin North Am 2000 Nov; 26 (4): 881-95; Bijlsma JW, Van den Brink HR. Am J Reprod Immunol 1992 Oct-Dec; 28 (3-4): 231-4; Jansson L, Holmdahl R .; Arthritis Rheum 2001 Sep; 44 (9): 2168-756; and Purdie DW. Br Med Bull 2000; 56 (3): 809-23; Merck Manual, 17th edition, pp. 449-451]. Aspects of the invention include methods of treating, reducing, inhibiting or preventing arthritis conditions, comprising administering a therapeutically effective amount of a composition of the invention. Another aspect relates to a method for treating, reducing, inhibiting or preventing osteoarthritis, comprising administering a therapeutically effective amount of a composition of the present invention.

As used herein, the term “bisphosphonate” corresponds to formula 1 below.

In the above formula,

R ₁ is independently selected from the group consisting of H, OH and Cl,

R ₂ is independently CH ₃ , Cl, CH ₂ CH ₂ NH ₂ , (CH ₂ ) ₃ NH ₂ , CH _2-3 -pyridyl, CH ₂ -S-phenyl-Cl, CH ₂ CH ₂ N (CH ₃₎ (cyclopentyl), CH ₂ - imidazole, CH ₂ -2- imidazo-pyridinyl, N- _{(cycloheptyl), CH 2 CH (CH 3} ) 2, (CH 2) 5 NH 2 and CH ₂ - 1-pyrrolidinyl and mixtures thereof. In an embodiment of the invention, R ₁ is OH and R ₂ is a 3-aminopropyl moiety such that the compound obtained is 4-amino-1-hydroxybutylidene-1,1-bisphosphonate, ie alendronate.

Pharmaceutically acceptable salts, derivatives and hydrates of bisphosphonates are also encompassed by the compounds and methods of the present invention. Non-limiting examples of salts include alkali, alkaline metal, ammonium and mono-, di-, tri- or tetra-C ₁ -C ₃₀ -alkyl substituted ammonium, including sodium, potassium, calcium, magnesium and ammonium salts. Selected from the group consisting of: Non-limiting examples of derivatives include those selected from the group consisting of esters and amides. Also included within the scope of this invention are various hydrates and other solvates of bisphosphonates, and pharmaceutically acceptable salts thereof. Hydrates of alendronate and their crystalline forms are included, including but not limited to within the scope of the present invention, hydrates having a water content of about 1-12%. Non-limiting examples of alendronate and other bisphosphonates include monohydrate, dihydrate, trihydrate, hemihydrate, quarter hydrate, 1/3 hydrate, 2/3 hydrate, 3/4 hydrate, 5/4 hydrate, 4/3 hydrate And 3/2 hydrates.

Non-limiting examples of bisphosphonates useful in the present invention include the following:

Allendronic acid, 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid.

Alendronate (also known as Alendronate sodium or monosodium trihydrate, trade name: FOSAMAX ), 4-amino-1-hydroxybutylidene-1,1-bisphosphonic acid monosodium trihydrate. (Alendronic acid and alendronate are described, for example, in US Pat. No. 4,922,007 to Kieczykowski et al., May 1, 1990 and US Pat. No. 5,019,651 to Kieczykowski et al., May 28, 1991). .

Cycloheptylaminomethylene-1,1-bisphosphonic acid, YM 175, Yamanouchi (Incadronate or Shimadronate) (e.g., described in US Pat. No. 4,970,335 to Isomura et al. On November 13, 1990) ).

1,1-dichloromethylene-1,1-diphosphonic acid (clodonic acid) and disodium salts (chlorodenate, Procter and Gamble) are described, for example, in Belgian Patent No. 672,205 (1966). And J. Org. Chem 32, 4111 (1967).

1-hydroxy-3- (1-pyrrolidinyl) -propylidene-1,1-bisphosphonic acid (EB-1053).

1-hydroxyethane-1,1-diphosphonic acid (ethidronic acid).

1-hydroxy-3- (N-methyl-N-pentylamino) propylidene-1,1-bisphosphonic acid (also known as BM-210955, Boehringer-Mannheim (ibandronate); for example US Patent No. 4,927,814, issued May 22, 1990).

[1-hydroxy-2-imidazopyridine- (1,2-a) -3-ylethylidene] -bis-phosphonate (minodronate).

6-amino-1-hydroxyhexylidene-1,1-bisphosphonic acid (neridronate).

3- (dimethylamino) -1-hydroxypropylidene-1,1-bisphosphonic acid (olpadronate).

3-Amino-1-hydroxypropylidene-1,1-bisphosphonic acid (pamideronate).

[2- (2-pyridinyl) ethylidene] -1,1-bisphosphonic acid (pyridronate) (see, eg, US Pat. No. 4,761,406).

1-hydroxy-2- (3-pyridinyl) -ethylidene-1,1-bisphosphonic acid (risdronate).

(4-Chlorophenyl) thiomethane-1,1-disphosphonic acid (tylideronate) (e.g., described in US Pat. No. 4,876,248 to Breliere et al. On October 24, 1989).

1-hydroxy-2- (1H-imidazol-1-yl) ethylidene-1,1-bisphosphonic acid (zoledronate).

In an embodiment of the invention, the bisphosphonates are selected from the group consisting of alendronates, pharmaceutically acceptable salts, derivatives and hydrates thereof and mixtures thereof. Pharmaceutically acceptable salts of alendronate may be selected from the group consisting of the sodium, potassium, calcium, magnesium and ammonium salts of allendronate and include allandronate monosodium or, for example, allandronate sodium monohydrate or allandronate sodium trihydrate. May be its hydrate.

In one embodiment, the compositions of the present invention comprise allendronate sodium (monosodium salt of 4-amino-1-hydroxybutylidene-1,1-bisphosphate), which is a member of the nitrogen-containing bisphosphonate group of the medicament.

As used herein in referring to a therapeutic agent of the present invention, the term “bisphosphonate (s)” includes diphosphonates, bisphosphonic acids and diphosphonic acids, as well as salts, derivatives and hydrates of these materials. it means. The use of specific nomenclature in referring to bisphosphonate (s) is not intended to limit the scope of the invention unless otherwise indicated. Because of the mixed nomenclature commonly used by those skilled in the art, the specific weight or percentage of bisphosphonate compound in the present invention is based on the acid active weight, unless otherwise indicated herein. Bisphosphonate doses calculated on the basis of their salt, derivative or hydrate forms are included within the dose range of the present invention based on their bisphosphonic acid active weight. In addition, the doses of all hydrate forms of the alendronate are calculated based on the active weight of the allandronic acid. For example, the monohydrate, trihydrate, hemihydrate, and all other hydrate forms of the alendronate and salts thereof are calculated based on their weight of the active action of alendronic acid. As another example, the sentence "about 70 mg of bone absorption inhibiting bisphosphonate selected from the group consisting of alendronate, pharmaceutically acceptable salts, derivatives and hydrates thereof, and mixtures thereof, based on the active weight of alendronic acid" is selected. It is meant that the amount of bisphosphonate compound is calculated based on 70 mg of alendronic acid.

As used in this specification and claims, the terms “bisphosphonic acid” and “alendronic acid” include the related bisphosphonic acid forms, their pharmaceutically acceptable salt forms, and equivalent mixtures thereof. The term includes crystals, hydrated crystals, and amorphous forms of alendronic acid and pharmaceutically acceptable salts thereof. The term “alendronic acid” in particular includes, but is not limited to, anhydrous alendronate monosodium, alendronate monosodium hemihydrate, alendronate monosodium monohydrate, alendronate monosodium trihydrate, anhydronate allandronate dipotassium and alendronate dipotassium pentahydrate do. Alendronate monosodium monohydrate and other crystalline forms of alendronate sodium are described in US Pat. No. 6,281,381. Potassium salts of alendronic acid and their hydrates are described in WO 99/20635.

While it is common to determine and calculate the dose of bisphosphonate based on the bisphosphonic acid active weight, the bisphosphonate dose can be calculated and administered based on other salt or hydrate forms. For example, the dosage of bisphosphonate risedronate is calculated based on the weight of anhydrous risedronate sodium salt. According to Physician's Desk Reference (55 ^th Edition, page 2654 (2001)), for example, each risedronate tablet is anhydrous risedronate in the form of a semi-pentahydrate with a small amount of monohydrate. It contains 5 to 30 mg of sodium.

As used herein, the term "cholecalciferol granules" as used in the collation means granules containing the knife during Ferrol and also vitamin D ₃ precursor, isomers of vitamin D _3, transesterification vitamin D ₃ or its isomers and / or It may contain additional excipients.

As used herein, the term “continuous schedule” or “continuous schedule of administration” means that the dosage regimen is repeated until the desired therapeutic effect is achieved. Continuous or continuous dosing schedules are distinguished from periodic or intermittent dosing.

As used herein, the term "dry vitamin D ₃ 100 granules" as used means that the commercially available gelatin-coated granules of the pharmaceutical grade dry vitamin D ₃ 100 from BASF.

As used herein, the term "universal bone loss" refers to bone loss through multiple skeletal sites or skeletal systems. The term "local bone loss" refers to bone loss of one or more specific defined skeletal sites.

As used herein, the terms “person in need of treatment”, “person in need of prevention”, “person in need of it” and “person at risk” refer to a disease state, as determined by a clinician or researcher. Means a person in need of treatment, prevention, alleviation, inhibition or reduction of a disease state, or at risk of developing a disease state.

As used herein, the term "IU" refers to International Units. In referring to the efficacy and dosage of vitamin D, it is customary to use international units (IU). One international unit (IU) is defined as the specific biological activity of crystalline international standard or 0.025 μg pure vitamin D. Stated another way, 1 μg of vitamin D is approximately 40 IU.

As used herein, the terms "mammal in need of treatment", "mammal in need of prevention", "mammal in need thereof" and "mammal at risk", as determined by a clinician or researcher, Means a mammal in need of treatment for a disease state, prevention, alleviation, inhibition or reduction of a disease state, or at risk of developing a disease state.

As used herein, the term “weekly administration” refers to a unit dose of bisphosphonate and a vitamin D compound once a week, ie once during a 7 day cycle, preferably on the same day each week. It means to administer to. In a weekly dosing regimen, the unit dose is generally administered about every seven days. A non-limiting example of a weekly dosing regimen is the administration of unit doses of bisphosphonates and vitamin D compounds every Sunday. It is commonly suggested that unit doses for weekly administration are not administered on consecutive days, but weekly dosage regimens may include dosage regimens in which the unit dose is administered two consecutive days within a different two week period. have.

As used herein, the term "osteoproliferator" refers to newly formed bone structures located around the joints, the occurrence of which is significantly associated with the later stages of osteoarthritis progression. The current hypothesis is that osteoblasts are produced from activated periosteum that induces new cartilage growth, which is actually converted into bone by the process of intrachondral bone formation.

As used herein with reference to salts, esters, hydrates, and derivatives of bisphosphonates (such as alendronates), the term “pharmaceutically acceptable” refers to the same general pharmacology as the free acid form from which the salts, derivatives, or hydrates of bisphosphonates are derived. Has the properties of a chemical and is acceptable in terms of toxicity.

As used herein, the term "pharmaceutically effective amount" means the amount of a compound, such as a bisphosphonate compound or vitamin D compound, that exhibits the desired therapeutic effect or response when administered according to a therapeutic diet. Pharmaceutically effective amounts of bisphosphonates are, for example, amounts administered in accordance with a therapeutic regimen sufficient to induce the prevention, reduction, inhibition or treatment of abnormal bone absorption.

As used herein, the term "pharmaceutical grade" means sufficient quality and efficacy to meet the introductory requirements of the applicable US Pharmacopoeia (USP) and the European Pharmacopoeia (Ph Eur). Nowadays, there are no USP studies on formulated vitamin D ₃ products, but the Ph. Eur. A research paper has been published. Cholecalciferol of "pharmaceutical grade" is, for example, a grade that is superior to vitamin D, which is generally used as a nutritional supplement.

As used herein, the term "preventing, inhibiting, reducing or treating" refers to abnormal bone uptake (and resulting physiological conditions such as osteoporosis) through direct or indirect changes in osteoclast formation or action. Prevention, inhibition, reduction or treatment of bone loss, in particular inhibition of removal of bone present from the mineral phase and / or organic matrix phase through direct or indirect changes in osteoclast formation or action. These terms are also meant to refer to other disease states or conditions in a manner that promotes alleviation from the disease or condition.

As used herein, the term “until the desired therapeutic effect is achieved” refers to the administration of a composition, such as a bisphosphonate and cholecalciferol composition, according to a selected dosing schedule, or where the course of treatment requires a disease or condition. Mean that the clinical or medical effect is repeated until the point at which it is observed by the clinician or researcher. In the method of treatment of the present invention, the bisphosphonate compound can be administered continuously until the desired change in bone mass or structure is observed. In this case, one of the objectives is to achieve an increase in bone mass, to prevent further reduction in bone mass, or to replace abnormal bone structures with more normal bone structures. In the method of the present invention, the bisphosphonate compound can be administered continuously only as long as necessary to prevent an undesired condition. In such cases, maintenance of bone mass density is often a goal. Non-limiting examples of administration periods can range from about 2 weeks to the remaining lifespan of the mammal. For humans, the administration period is from about 2 weeks to the rest of the human life, preferably from about 2 weeks to about 40 years, more preferably from about 1 month to about 35 years, more preferably from about 6 months to about 30 Years and most preferably about 1 year to about 20 years.

As used herein, the term “vitamin D” refers to both vitamin D ₂ and vitamin D ₃ , which have the formula shown in FIG. 3. As used herein, the terms "metabolites of vitamin D" and "derivatives of vitamin D" refer to vitamin D ₂ and metabolites and derivatives of vitamin D ₃ . As used herein, the term “vitamin D compound” refers to vitamin D ₂ (ergocalciferol) and vitamin D ₃ (cholecalciferol), 7-dehydrocholesterol and vitamin D ₂ precursor and vitamin D ₃ precursor, 7-dehydrocholesterol, vitamin D ₂ , vitamin D ₃ , vitamin D ₂ precursor or isomer or ester of vitamin D ₃ precursor, or mixtures thereof. At the relevant physiological temperature, vitamin D ₂ and vitamin D ₃ are identical to the isomers of their respective vitamin precursors, although the equilibrium shifts towards vitamin D ₂ and vitamin D ₃ . In the present invention, the term "vitamin D compound" does not include metabolites of vitamin D (such as 25-hydroxycholecalciferol or calcitriol, or their analogs), and the term also includes the active hormone calcitriol or its analogs. I never do that. The terms vitamin D ₃ and cholecalciferol are used interchangeably unless otherwise indicated.

The present invention provides a composition comprising bisphosphonates, or pharmaceutically acceptable salts, derivatives or hydrates of bisphosphonates, or mixtures thereof and vitamin D compounds. In an exemplary embodiment, the bisphosphonate compound is selected from the group consisting of Alendronate Sodium, Alendronate Sodium Monohydrate, and Alendronate Sodium Trihydrate, and the vitamin D compound is cholecalciferol.

The exact dose of bisphosphonate and vitamin D compound will depend on the schedule of administration, the oral efficacy of the particular bisphosphonate chosen, the age, size, sex and condition of the mammal or human, the nature and severity of the disease to be treated, and other related medical and physical factors. . Thus, the exact pharmaceutically effective amount cannot be defined in advance, but can be easily determined by a caregiver or clinician. Appropriate amounts can be determined by routine experimentation from animal models and clinical studies in humans. In general, the pharmaceutically effective amount of bisphosphonate is selected according to the dosing schedule that continues until the desired therapeutic effect is achieved. In humans, an oral dose effective amount of bisphosphonate is typically from about 0.0001 to about 100 mg / kg body weight, preferably from about 0.0005 to about 20 mg / kg body weight for a subject weighing 75 kg.

In an embodiment of the present invention, an appropriate amount of vitamin D compound is selected to provide adequate vitamin D nutrition during the interval of administration without interfering with the ability of bisphosphonates to achieve a bone resorption inhibitory effect. For oral compositions of the invention comprising alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or a mixture thereof and vitamin D, the amount of vitamin D compound comprises from about 100 to about 60,000 IU. In embodiments of the present invention, non-limiting examples of oral doses of vitamin D compounds include, but are not limited to, vitamin D compounds 700 IU, 1,400 IU, 2,800 IU, 4,200 IU, 5,600 IU, 7,000 IU, 8,400 IU, 14,000 Capacities of IU, 28,000 IU, 36,000 IU and 60,000 IU.

In the case of an oral composition comprising a vitamin D compound and a pharmaceutically effective amount of alendronate or a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof, the oral pharmaceutically effective amount of the alendronate is typically the weight of alendronic acid. Based on about 0.05 to about 1120 mg of the alendronate compound. Non-limiting examples of oral pharmaceutically effective amounts of alendronate in embodiments of the invention are not limited thereto, but are about 2.5 mg, 5 mg, 8.75 mg, 10 mg, 17.5 mg, 35, based on the weight of the alendronic acid, respectively. Alendronate doses of mg, 40 mg, 70 mg, 140 mg, 280 mg, 560 mg and 1120 mg.

Bisphosphonates and vitamin D compositions of the invention are typically administered with a suitable pharmaceutical diluent, excipient or carrier appropriately selected for the dosage form for oral administration. Examples of oral dosage forms include tablets (including compressed, coated or uncoated), capsules (each of which include sustained release formulations or timed release formulations), hard or soft gelatin capsules, pellets, pills, powders, Granules, elixirs, tinctures, slurries, foaming compositions, films, sterile solutions or suspensions, syrups and emulsions and the like. Similarly, it is also an intravenous (temporary injection or infusion), intraperitoneal, topical (eg ophthalmic eye drops), nasal, inhalation, subcutaneous, intramuscular or transdermal use of all forms well known to those skilled in the pharmaceutical arts. (Eg, in a patch) form, metered aerosol or liquid spray, drop, ampoule, self-injecting device or suppository. Non-toxic effective amounts of the desired compositions can be used. The composition is suitable for oral, parenteral, nasal, sublingual or rectal administration, or administration by inhalation, insufflation. Compositional formulations according to the invention are described, for example, in Remington's Pharmaceutical Sciences, 17 ^th ed. It may conveniently be carried out by methods known in the art as described in 1995).

For example, when administered orally in the form of tablets, capsules, pellets or powders, the active ingredient is an oral, nontoxic, pharmaceutically acceptable inert carrier such as lactose, starch, sucrose, glucose, methyl Cellulose, magnesium stearate, mannitol, sorbitol, croscarmellose sodium, etc.) and orally administered in liquid form (e.g., elixirs, syrups, slurries, emulsions, suspensions, solutions and foaming compositions) In this case, the oral pharmaceutical ingredient may be mixed with an oral nontoxic pharmaceutically acceptable inert carrier such as ethanol, glycerol and water. Moreover, if desired, suitable binders, fillers, diluents, lubricants, compression aids, disintegrants, buffers, coatings and coloring agents may also be incorporated. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose, anhydrous lactose, free flowing lactose, beta-lactose and corn sweeteners, natural and synthetic rubbers such as acacia, guar, trad. Gancant or sodium alginate), carboxymethyl cellulose, polyethylene glycol and wax, and the like. Lubricants used in these dosage forms can include but are not limited to sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Suitable disintegrants may be one of several modified starch or modified cellulose polymers, including croscarmellose sodium. Diluents that can be used as compression aids include, but are not limited to, lactose, dicalcium phosphate, cellulose, microcrystalline cellulose, and the like. Glidants that improve the flow characteristics of the powder mixture can also be used in the present invention. Examples of glidants include, but are not limited to, colloidal silicon dioxide and talc. The compositions used in the methods of the invention may also be combined with soluble polymers as the desired pharmaceutical carrier. Such polymers may include, but are not limited to, polyvinylpyrrolidone, pyran copolymers, polyhydroxypropyl-methacrylamide, and the like. Additional excipients as described in US Pat. Nos. 5,358,941, 5,882,656 and PCT Publication WO 95/29679 may also be used.

One aspect of the invention corresponds to, for example, about 0.5 to about 90 weight percent of alendronate sodium, about 1 to about 70 weight percent of cholecalciferol granules (about 0.0005 to about 20 weight percent of cholecalciferol) ), About 10 to about 80 weight percent anhydrous lactose, about 5 to about 50 weight percent microcrystalline cellulose, about 0.1 to about 5 weight percent colloidal silicon dioxide, about 0.5 to about 10 weight percent croscarmellose 80 to 1500 mg tablets comprising sodium and about 0.5 to about 5 weight percent magnesium stearate.

The weight range of 100 g of anhydrous vitamin D ₃ granules is to guarantee the efficacy of 2800 IU in each tablet, since the granules contain an effective range of 100,000 to 110,000 IU of vitamin D ₃ per gram. The amount of anhydrous lactose is adjusted in accordance with the amount of anhydrous vitamin D ₃ 100 granules added to the tablet so that the final total weight is maintained at 325 mg. Other non-limiting examples of oral compositions of the invention comprising bisphosphonate compounds (such as alendronate) and vitamin D compounds are described herein, including the following examples. Anhydrous Vitamin D ₃ 100 granules contain about 100,000 IU of vitamin D ₃ per gram of granule weight. Thus, 28 mg of anhydrous vitamin D ₃ 100 granules contain about 2800 IU of vitamin D ₃ , which is equivalent to about 70 μg of vitamin D ₃ .

Including the embodiments described herein, the bisphosphonate / vitamin D compositions of the invention can be administered weekly, two weeks, one month, two times a month, and two month intervals. For week-to-week administration with the compositions of the present invention, the oral pharmaceutically effective amount of the alendronate comprises from about 0.05 to about 1120 mg of the alendronate compound, based on the weight of the alendronic acid active. Embodiments of the invention that provide a weekly oral pharmaceutically effective amount of alendronate include, but are not limited to, unit doses useful for the prevention of osteoporosis comprising a vitamin D compound and about 35 to about 70 mg of alendronate compound; Unit doses useful for the treatment of osteoporosis, including vitamin D compounds and about 70 mg of alendronate compounds; Unit dose useful for treating Paget's disease, including a vitamin D compound and about 280 mg of an alendronate compound; And unit doses useful for the treatment of metastatic bone disease, including vitamin D compounds and about 280 mg of alendronate compounds.

For weekly administration, the pharmaceutically effective amount of the vitamin D compound in the bisphosphonate / vitamin D composition of the invention comprises about 100 to about 60,000 IU of vitamin D. Thus, in an aspect of the invention, the composition comprises about 100 to about 5600 IU of vitamin D compound and a pharmaceutically effective amount of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof. In another embodiment, the pharmaceutically acceptable amount of the alendronate comprises from about 0.05 to about 1120 mg of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or a mixture thereof, based on the alendronic acid activity.

For two weeks or two months of administration, the pharmaceutically effective amount of the vitamin D compound in the bisphosphonate / vitamin D composition of the invention comprises about 100 to about 60,000 IU of vitamin D. In an embodiment of the invention, the composition comprises about 100 to about 8,400 IU of vitamin D compound and a pharmaceutically effective amount of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof. In another embodiment, the pharmaceutically acceptable amount of the alendronate comprises from about 0.05 to about 1120 mg of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or a mixture thereof, based on the alendronic acid activity.

For monthly administration, the pharmaceutically effective amount of the vitamin D compound in the bisphosphonate / vitamin D composition of the present invention comprises about 100 to about 36,000 IU of vitamin D. In one aspect of the invention, the composition comprises about 100 to about 11,200 IU of vitamin D compound and a pharmaceutically effective amount of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or mixtures thereof. In another embodiment, the pharmaceutically acceptable amount of the alendronate comprises from about 0.05 to about 1120 mg of alendronate, a pharmaceutically acceptable salt, derivative or hydrate thereof, or a mixture thereof, based on the alendronic acid activity.

The invention also includes the prevention, reduction, inhibition and treatment of diseases and conditions (eg osteoporosis) associated with abnormal bone absorption. Administration of a composition of the present invention according to the method of the present invention in a person suffering from osteoporosis, i.e., having a standard deviation of bone mineral density (BMD) of less than or equal to about 2 or 2 1/2 or more below the norm of postmenopausal women. Supported. Vitamin D ₃ administered once a week up to 7 times or more of the amount provided on a daily basis does not adversely affect the bioavailability of bisphosphonates and may be administered simultaneously with bisphosphonates (eg, alendronate) (Examples 7). The methods of the present invention do not suffer from the disadvantages of current therapies which require annoying, irregular or complex dosage regimens to provide adequate vitamin D during bisphosphonate treatment.

As a result, a composition of the present invention, such as a composition comprising a bisphosphonate compound (such as alendronate) and a vitamin D compound, is effective for all guidelines in which a composition comprising an alendronate or other bisphosphonates without a vitamin D compound is effective. The methods and compositions of the present invention are useful for reducing or inhibiting bone absorption and for treating, reducing, inhibiting or preventing abnormal bone absorption and conditions associated therewith. Accordingly, the compositions of the present invention can be used in humans and other animals to increase bone mass, prevent, inhibit, reduce and treat the following conditions and disease states: bone loss; Osteoporosis, including but not limited to postmenopausal osteoporosis, steroid induced osteoporosis, male osteoporosis, disease induced osteoporosis, idiopathic osteoporosis and glucocorticoid induced osteoporosis; Bone necrosis, Paget's disease; Osteoarthritis; Rheumatoid arthritis, other arthritic conditions, abnormally increased bone turnover; Local bone loss or osteolysis associated with periprostate bone loss; Bone fractures; Metastatic bone disease; Gaucher's disease; Avascular necrosis; Polyfibrous dysplasia; Charcot's joint; Parasitic diseases; Osteopenic insufficiency; Homocystinuria; carbohydrate Metabolic abnormalities; Turner's syndrome; dissimilation; Advanced osteoplastic fibrous dysplasia; Osteoincomplete fibrosis; Periodontal disease; Tooth loss; Hypercalcemia of malignant tumors; Multiple myeloma; Thereby, but not limited to, osteopenia, including immobilized induced osteopenia and osteopenia due to bone metastasis and other bone diseases and conditions that may be associated with abnormal bone resorption.

The present invention relates to the use of a composition of the invention for the manufacture of a medicament useful for the treatment, reduction, inhibition or prevention of arthritis conditions. The invention also relates to compositions of the invention and inhibitors of androgen receptor modulators, osteoclast proton ATPases for the preparation of medicaments useful for the treatment of arthritis conditions; Inhibitors of HMG-CoA reductase; Osteoclast assimilation agents; Calcitonin; The use of an agent selected from the group consisting of vitamin K ₂ or a pharmaceutically acceptable salt thereof and mixtures thereof.

In one embodiment of the present invention, the arthritis condition includes amyloidosis, ankylosing spondylitis, bacterial arthritis, basic calcium phosphate crystal deposition disease; Behcet's disease, bursitis and tendonitis, CPPD deposition disease, calcific tendinitis, carpal tunnel syndrome, Elus-danlos syndrome, enteropathic arthritis, Pelti syndrome, myalgia, gout, fungal arthritis, hemochromatosis, hematopoietic arthritis, hypertrophic Osteoarthritis, Infectious Arthritis, Inflammatory Bowel Disease, Combustible Arthritis, Combustive Rheumatoid Arthritis, Lupus Erythematosus, Lyme Disease, Marfan's Syndrome, Mixed Connective Tissue Disease, Multiple Cardiomyopathy, Myopathy, Myositis, Osteoarthritis, Bone Necrosis, Bone Necrosis Chondroitinosis , Polyarthritis, rheumatoid polymyalgia, psoriatic arthritis, Raynaud's phenomenon, reflex sympathetic dystrophy syndrome, Radar syndrome, recurrent polychondritis, rheumatoid arthritis, rheumatic fever, sarcoidosis, septic arthritis, scleroderma, jogren Syndrome, vertebral dysplasia, systemic lupus erythematosus and viral arthritis.

One aspect of the invention relates to a method for treating, reducing, inhibiting or preventing the progression of osteoarthritis in a mammal in need thereof comprising administering to the mammal a therapeutically effective amount of a composition of the invention. It is known in the literature that osteoarthritis involves well defined changes in joints, including erosion of articular cartilage surfaces, periarticular cartilage / osteobacteria, zinc bone osteonecrosis, and cyst formation [Oettmeier, R. Abendroth, K, "Osteoarthritis and bone: osteologic types of osteoarthritis of the hip", Skeletal Radiol. 1989; 18: 165-74. Recently, the potential contribution of zinc bone bone necrosis to the onset and progression of osteoarthritis has been suggested. Rigid zinc bone, such as joints that respond to repeated seizure loads, cannot weaken and disperse forces through the joints, resulting in greater mechanical stress applied through the articular cartilage surface. This in turn promotes cartilage wear and fibrillation [Radin, EL and Rose RM ", Role of subchondral bone in the initiation and progression of cartilage damage", Clin. Orthop. 1986; 213: 34-40. Excessive suppression of osteoarticular bone uptake by the compositions of the present invention will lead to inhibition of zinc bone turnover, which may have a useful effect on osteoarthritis progression.

Another aspect of the invention relates to a method for treating, reducing, inhibiting or preventing a rheumatoid arthritis condition in a mammal in need thereof comprising administering to the mammal a therapeutically effective amount of a composition of the invention. The literature discloses that progressive destruction of periarticular bone is a major cause of joint dysfunction and asthenia in patients suffering from rheumatoid arthritis [Ref. Goldring SR, "Pathogenesis of bone erosions in rheumatoid arthritis", Curr. Opin. Rheumatol, 2002; 14: 406-10. In addition, universal bone loss is a major cause of mobility associated with severe rheumatoid arthritis. The frequency of hip and spinal cord fractures is substantially increased in patients with chronic rheumatoid arthritis [Refer to Gould A, Sambrook, P, Devlin J et al., "Osteoclastic activation is the principal mechanism leading to secondary osteoporosis in rheumatoid arthritis ", J. Rheumatol. 1998; 25: 1282-9]. The use of anti-absorbents in the absorption of osteoarticular bone and the treatment or prevention of universal bone loss represents a rational approach of pharmacological intervention to the progression of rheumatoid arthritis. Thus, compositions of the present invention comprising bisphosphonate compounds and vitamin D compounds can be used to treat, reduce, inhibit or prevent bone loss associated with rheumatoid arthritis and other osteoarthritis conditions.

More generally, it is believed that bisphosphonates can be provided in the same formulation with vitamin D without adversely affecting the bioavailability of bisphosphonates. Moreover, it is believed that lower doses of vitamin D and higher doses of vitamin D can be provided in the same formulation without adversely affecting their bioavailability. As an example, weekly administration of 2800 IU vitamin D is considered effective when administered with bisphosphonates in the compositions of the present invention. In addition, there is a vitamin D ₂ may be used instead of vitamin D _3, which has a similar result as that found in vitamin D _3. Thus, administration of a vitamin D compound in the same formulation with a bisphosphonate compound eliminates the separate dosing requirement of vitamin D during bisphosphonate treatment and provides vitamin D nutrition without adversely affecting the bioavailability and efficacy of the bisphosphonate.

Patients benefit from a mixture of vitamin D and bisphosphonates because they provide additional vitamin D nutrition to facilitate bone formation and mineralization and improve the efficacy of bisphosphonate treatment. In terms of the patient's lifestyle and practice, the methods of the present invention are also more convenient than daily or periodic dosing regimens for bisphosphonates in addition to daily vitamin D administration. As a result of the present invention, because the present invention provides weekly administration of vitamin D, the patient also no longer needs to take vitamin D separately each day so that additional vitamin D nutrition is useful. Patients do not have to follow the complex dosing regimen of separate bisphosphonates and vitamin D administrations. Finally, patients will be less likely to take the bisphosphonate compound in the empty stomach and to fast for 30 minutes or more, either before or after administration. Thus, the methods of the present invention may have the advantage of facilitating better patient performance and thus can be represented as useful therapeutic efficacy.

It is also believed that bisphosphonate / vitamin D compositions are less irritating to the esophagus and gastrointestinal system. Alendronate can potentially penetrate into the basal layer of the stratified squamous epithelium (e.g., through its own infiltration or through infiltration into localized damage sites caused by wearable foods or other agents). As suggested by its effect on growth, it can lead to inhibition of keratinocyte growth. See AA Reszka et al., Mol. Pharmacol., 2001; 59 (2): 193-202. Growth inhibition can slow the epithelial repair process, leading to local irritation or ulceration. Autoradiograms of rats fed with radioactive 1,25 (OH) ₂ vitamin D ₃ show evidence of the expression of vitamin D receptors in the epithelium of the esophagus (see Stumpf, WE, et al., Histochemistry, 1987; 87 (1): 53-8]. The level is less than that seen in the parathyroid glands.

Thus, co-administration of active vitamin D ₃ metabolites (eg, 1,25 (OH) ₂ -cholecalciferol or calcitriol) with alendronate is believed to exhibit esophageal irritation through its undifferentiated effect on keratinocytes. . Both calcitriol and 25-OH-cholecalciferol have been observed to affect keratinocytes by inhibiting growth and inducing differentiation. K. Matsumoto et al., Biochem. Biophys. Acta., 1991; 1092 (3): 311-8]. Because keratinocyte differentiation is associated with cell cycle arrest (growth arrest), a mixture of allendronate and active vitamin D metabolites can have a synergistic effect on inhibiting growth at the bed base. This may eventually lead to greater stimulant effects. It is believed that oral vitamin D ₃ (cholecalciferol) does not have the same local stimulant effect as the active metabolite because it requires activation in the liver followed by kidney.

In addition, normal physiological levels of the active vitamin D ₃ hormone can assist the body in its attempts to repair local irritant sites induced by acute exposure to the alendronate. Oral vitamin D ₃ is administered in combination with alendronate, and since the majority of the older population are vitamin D ₃ deficient, it can be better faster in the process of healing the body.

Vitamin D / bisphosphonate compositions are also considered useful for the prevention or treatment of sway. In addition, vitamin D / bisphosphonate compositions are believed to be useful for reducing falls. Vitamin D / bisphosphonate compositions are believed to increase muscle strength, improve neuromuscular function, reduce body sway, and improve physical function in adults. This leads to a reduced risk of poles, thus contributing to a reduced risk of bone fractures. Epidemiologic studies have shown a large prevalence of vitamin D deficiency in US adults [Ref. A.N. Exton-Smith et al., Lancet 1966; 2: 999-1001; RP Heaney et al., Osteoporos Int 2000; 11: 553-5; MJ McKenna, Am J Med 1992; 93: 69-77; ST Haden et al., Calcif Tissue Int 1999; 64: 275-9. There is evidence of the effects of vitamin D on extra-skeletal tissues (Latham et al, 2003; 51: 1219-1226). In addition, vitamin D receptors have been identified in muscle tissue, and muscle weakness, limb pain and impaired physical function are well recognized as signs of severe vitamin D deficiency.

Numerous prospective randomized intervention studies have shown the efficacy of vitamin D in improving musculoskeletal function and reducing the risk of poles. Treatment with vitamin D and calcium has been shown to reduce the incidence of non-vertebral fractures and reduce the incidence of posture sway and possibly poles. JK Dhesi et al., Age and Aging 2002; 31 : 267-271]. In addition, the number of poles in adult group-resident patients has been found to be significantly reduced by treatment with alphacalcidol (1-alpha-hydroxyvitamin D ₃ ) and minimal calcium intake [L. Dukas et. al., JAGS 2004; 52: 230-236.

Vitamin D / bisphosphonate compositions are also believed to improve absorption of calcium. There has been a study using the active metabolite of vitamin D to observe the positive effects on partial calcium absorption in postmenopausal women [Ref. M.L. Holzherr et al., Osteoporosis Int 2000; 11: 43-51; J.C. Gallagher et al., J. Clin Endocrinol. Metab., 1980; 51 (5): 1359-64]. Bisphosphonates have also been found to increase intestinal calcium uptake in rat models [P. Ammann et al., J Bone Miner Res 1993; 8 (12): 1491-8; H. Fleisch Osteoporos Int 1996; 6: 166-70; J.P. Bonjour Endocrinol Metab 1988; 17: E260-E264]. However, compositions of vitamin D compounds and bisphosphonates are believed to increase calcium uptake over the additive effects of vitamin D or bisphosphonates administered alone, respectively. In addition, cholecalciferol and alendronate compositions are believed to improve calcium absorption more than combinations of vitamin D in the active form and different bisphosphonates. This increase in calcium absorption correlates with a reduction in fracture risk.

The invention also provides a pharmaceutically acceptable form of a vitamin D compound for the manufacture of a medicament for the treatment, reduction, inhibition or prevention of the above mentioned conditions and diseases in a mammal (eg a human) and a pharmaceutically effective amount of at least one bisphosphonate or bisphosphonate. Provided are bisphosphonate compounds comprising salts, derivatives or hydrates or mixtures thereof, and compositions comprising one or more active ingredients.

In another embodiment, the methods and compositions of the present invention may also include histamine H2 receptor blockers (ie antagonists) and / or proton pump inhibitors, which are well known as therapeutic agents that increase the pH of the stomach [ References: LJ Hixson et al., Current Trends in the Pharmacotherapy for Peptic Ulcer Disease, Arch. Intern. Med., Vol. 152, pp. 726-732 (April 1992). It has been found in the present invention that sequential oral administration of bisphosphonates and vitamin D compositions, followed by histamine H2 receptor blockers and / or proton pump inhibitors, may help to minimize gastrointestinal adverse effects. In one embodiment of the invention, the histamine H2 receptor blocker and / or proton pump inhibitor is administered from about 30 minutes to about 24 hours, or from about 30 minutes to about 12 hours before the bisphosphonate and vitamin D composition. The dose of the histamine H2 receptor blocker and / or the proton pump inhibitor depends on the particular compound selected and the factors relevant to the mammal to be treated, i.e. size, health and the like. Non-limiting examples of histamine H2 receptor blockers and / or proton pump inhibitors include those selected from the group consisting of cimetidine, pamotidine, nizatidine, ranitidine, ohmprazole and lansoprazole.

The present invention also encompasses a method of making a composition of the present invention comprising, for example, a pharmaceutical composition comprising a bisphosphonate compound and a vitamin D compound. In one embodiment, a process for preparing an alendronate-cholecalciferol formulation comprises preparing a powder mixture comprising an alendronate, densifying the powder mixture to form an alendronate mixture, pulverizing and mixing the alendronate mixture and cholecalciferol granules Forming, and smoothing and compacting the mixture. In another embodiment, the process for preparing the alendronate-cholecalciferol solid dosage form comprises mixing alendronate, colloidal silicon dioxide, anhydrous lactose, microcrystalline cellulose, and croscarmellose sodium to form a preliminary mixture, and the premix and magnesium stearate To form a first lubricity mixture, compacting the first lubricity mixture to form a densified ribbon, pulverizing the densified ribbon to form a lubricity mixture, and mixing the lubricity mixture and cholecalciferol granules Forming a lubricity mixture, and then compressing the second lubricity mixture into a solid dosage form.

4 shows an embodiment flu-chart of the method of the present invention for preparing the bisphosphonate / vitamin D composition of the present invention. In this embodiment, the composition is prepared by a process comprising roller compacting the Alendronate sodium formulation to form a ribbon, pulverizing the ribbon prepared from the roller compaction step, and then mixing with extragranular addition of the vitamin D ₃ formulation. do. For example, using the active ingredients and excipients set forth in Example 1, this formulation and process produce a product that meets the regulatory requirements for product release and stability of both alendronate and vitamin D ₃ . As shown in FIG. 4, in this embodiment, in step 301, a premix of colloidal silicon dioxide, lactose anhydrous and alendronate sodium is prepared. As shown by step 302, the preliminary mixture is then mixed with microcrystalline cellulose and croscarmellose sodium. In step 303, magnesium stearate is added to form a lubricity mixture. The lubricious mixture is passed through a roller compactor as shown in step 304 and the densified ribbon is milled. In the embodiment shown in Figure 4, in step 305, about 2800IU (or the same as about 70㎍) it was added to Vitamin D ₃ Granule containing vitamin D _3, and mix the milled granules, and vitamin D ₃ are granular charge roller Adjustments are made based on both granulation analysis and yield introduced from the densification / grinding step. The resulting mixture is then compressed in step 306 to form tablets, and the compressed tablets remove dust. The resulting tablets may be packaged in suitable packaging including, for example, moisture-proof and shading foam packs or bottles.

Vitamin D ₃ (cholecalciferol) and vitamin D ₂ (ergocalciferol) are water insoluble hydrophobic compounds having a melting point of about 84 ° C. and about 115 ° C., respectively. These compounds are also very susceptible to oxidation and are prone to change by light, resulting in decomposition into various degradation products. Vitamin D granules are also easy to separate. Thus, the stability of vitamin D is influenced by the storage conditions (eg exposure to light, high temperatures and high relative humidity) of the vitamin D / bisphosphonate composition, as well as the degree and nature of processing. As a result, the purpose of including vitamin D in the composition of the present invention means a particular challenge as far as it relates to the development of the preparation method and the storage of the vitamin D-containing composition. Thus, there is a need for vitamin D / bisphosphonate compositions formulated to reduce degradation of vitamin D, both during processing and during storage. There is also a need for a method for producing such a stable composition. There is also a need for development of methods for detecting or measuring the degradation of vitamin D in vitamin D containing compositions such as the compositions of the present invention. In addition, since the level of a particular vitamin D breakdown agent may be very low (by nanograms), such as a composition of the present invention having a limit of quantitation (LOQ) sufficient to detect the amount of vitamin D breakdown agent, There is a need for the development of methods for measuring or detecting the degradation of vitamin D in vitamin D containing compositions.

Accordingly, the present invention also provides a method for preparing a composition comprising a bisphosphonate compound and a vitamin D compound that minimizes the loss of the vitamin D compound during manufacture. By adjusting the humidity, temperature and light present in the vitamin D formulation during manufacture, or by providing appropriate processed dosage form packaging, the loss of vitamin D can be reduced while maintaining its efficacy completely. In one embodiment of the invention, the temperature during the manufacturing process is about 35 ° C or less. In another embodiment, the temperature is about 20 to about 30 ° C. In one embodiment, the relative humidity during the manufacturing process is about 60% RH or less. In another embodiment, the relative humidity is about 20 to about 40%. In addition, the starting moisture level of the vitamin D component of the formulation can be adjusted, as well as excipients that may be present. In another embodiment, the relative humidity is about 25 to about 35%.

The present invention also encompasses a manufacturing method comprising an additional drying step. Thus, in another aspect, the compositions of the present invention can be prepared under different conditions (temperature and / or relative humidity) as described above, and the water content of the prepared composition can be reduced by drying the composition. In an aspect, drying may comprise drying (eg, using heat) the composition of the invention after the solid dosage form has been produced. In another embodiment, drying may comprise film coating a solid formulation (eg, a tablet) of the composition of the present invention. In other embodiments, drying may also include packaging the composition of the present invention with an appropriate amount of desiccant or other moiety to reduce the water content. In another aspect, drying may comprise storing the composition of the present invention in a storage form (eg, aluminum foam pack, moisture proof bottle) that reduces moisture and / or light.

In an embodiment of the invention, the vitamin D compound used as starting material may comprise stabilized granules that are free flow of vitamin D. In an embodiment, the vitamin D granules used as starting materials in the preparation method of the invention are pharmaceutical grade gelatin coated anhydrous vitamin D ₃ 100 from BASF. Vitamin D particles are dissolved in heavy chain triglycerides, which are droplets of 1-2 μm incorporated into a starch coated matrix of gelatin and sucrose. The dissolved vitamin D can then be stabilized with tertiary butylhydroxytoluene (BHT). Vitamin D granules contain sodium aluminum silicate as flow aid. Those skilled in the art will appreciate that the amount of vitamin D added to the composition needs to be adjusted based on the vitamin D source and / or the efficacy of the added vitamin D. For example, if pharmaceutical grade gelatin coated anhydrous vitamin D ₃ 100 granules (BASF) are used, the skilled person may have different efficacy of the granules (e.g. 100,000 IU / g or 105,000 IU / g or 110,000). IU / g), it will be appreciated that the amount of granules added to the composition must be adjusted to achieve vitamin D of 2800 IU or 5600 IU in the composition.

Vitamin D contained in the embodiment of the present invention is Ph. Eur. Cholecalciferol concentrate (powder form) Follow the permissible range of the research paper. At this time, there is no USP research paper for formulated vitamin D ₃ products, but the applicable Ph. Eur. The research paper was published. Inactive ingredients (such as medium chain triglycerides, butylated hydroxytoluene, sucrose, gelatin, modified starch and sodium aluminum silicate) in the compositions of the vitamin D compounds used in the composition embodiments of the invention are summarized or food grade materials to be.

In an embodiment of the methods and compositions of the present invention, the alendronate used as the starting material is the outlined grade of alendronate sodium monohydrate or the summarized grade of alendronate sodium trihydrate (Merk & Co., Inc.).

In addition, commercially available vitamin D granules, such as those sold by Roche, BASF or Solvay, can be used in the compositions of the present invention.

In another aspect, the present invention provides a kit for conveniently and effectively carrying out the method according to the present invention. Such kits are particularly suitable for the delivery of oral solid forms such as tablets or capsules, in which embodiments include cards having a number of unit doses, doses produced according to their intended use. An example of such a kit is a "foam pack." Foam packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dose forms. If desired, memory aids may be provided, for example, in the form of numbers, letters, or other indications, or with calendar inserts dated to the treatment schedule in which the dose may be administered. Alternatively, placebo doses or calcium or dietary supplements, in forms similar or distinct from bisphosphonates and vitamin D unit doses, may be included to provide a kit in which the dose is administered daily. In embodiments comprising histamine H2 receptors and / or proton pump inhibitors, these agents may be included as part of a kit.

The present invention also provides a detection method developed for measuring the degradation products of the vitamin D ₃ compounds of the present invention. In particular, a method of measuring degradation of a pharmaceutical composition comprises extracting cholecalciferol from the composition into a first solution to form a second solution, separating the sample containing cholecalciferol from the second solution, and then May be subjected to reverse phase HPLC separation to detect the amount of cholecalciferol in the sample. The detection method of the present invention is performed to detect from about 2800 to about 5600 IU of cholecalciferol per pharmaceutical composition. In addition, the detection method has a limit of quantitation (LOQ) of cholecalciferol of less than about 9 ng / ml cholecalciferol.

In one embodiment, the method uses a first solution comprising water, alcohol, acetonitrile or mixtures thereof. In a specific embodiment, the first solution contains about 5% water and about 95% methanol. An exemplary sample formulation can be extracted from 15 tablets containing 2800 IU of vitamin D with about 5% water and about 95% methanol about 50 ml, respectively. In addition, in one embodiment, the resulting solution can be stirred for about 10 minutes, sonicated for about 30 minutes, and then again for 3 hours. In one embodiment, separation of the sample may be performed by centrifugation, which may be between about 5,000 and about 15,000 rpm. In one embodiment, the column is a Phenomenex Phenosphere 80 mm ODS (1) column (150 × 4.6 mm, 3 μm) and the injection volume is 100 μl. Samples are cleated down the column and then detected. In an embodiment of the method, a 65 minute gradient can be used. Detection wavelengths of about 260 to about 265 nm can also be used. In one embodiment of the method, the detecting step is performed at a reverse phase HPLC column temperature of about 25 ° C. A sample tray temperature of about 5 ° C. may be used. In one embodiment, the detecting step comprises reverse phase HPLC separation using an eluent of about 99% acetonitrile and about 1% 0.025% phosphoric acid.

In one embodiment, reversed phase HPLC columns that may be used in the methods of the present invention include only partially or non-end capped columns. The end capped process reduces the free silanol groups on the stationary phase, thus performing separation between the vitamin D precursor and the vitamin D peak. Attempts to use end capped columns have not been successful in providing sufficient peak degradation for the assays of the present invention because the specific degradation products eluted between the two actives are not separated and quantified. Indeed, in identifying the columns for using the methods of the present invention, vitamin D ₃ isomers (0.96 RRT) were observed to elute between the two active agents found in the formulation. Larger developments of methods using other end capped columns all exhibit limited degradation between the vitamin D ₃ precursor and the vitamin D ₃ peak.

Column carbon loading is the product of transesterification between vitamin D ₃ precursors or vitamin D ₃ and medium chain triglycerides (C ₈ and C ₁₀ fatty acid esters are present as BASF vitamin D granules that can be used in the compositions and methods of the present invention). Affects the dissolution of the four vitamin D ₃ ester adducts. These esters can react with the hydroxy groups of vitamin D ₃ through transesterification mechanisms to form C ₈ -D ₃ and C ₁₀ -D ₃ esters. Due to the long chain fatty acids, these esters are very hydrophobic and interact with the C ₁₈ stationary phase. Columns with higher carbon loadings have more C ₁₈ stationary phases, thus interacting more strongly with esters and retaining esters on the columns for longer. Thus, in one embodiment of the method of the invention, an HPLC column with a carbon load of less than about 10% can be used. The use of a lower carbon loading column reduces the interaction between the stationary phase and the ester, leading to earlier elution of these peaks. From the results, for example, using a low carbon loading (5%) Platinum EPS C ₁₈ column, all esters are eluted 10 minutes before a mobile phase containing 95% acetonitrile / 5% water is used. Similarly, all four esters eluted within 26 minutes when a different column, Phenosphere ODS (1) column (7% carbon loading) was used.

Exemplary chromatography conditions that can be used in the methods of the present invention are given below.

Using the detection method of the present invention, both the pre-vitamin D and vitamin D peaks can be quantified to calculate the total amount of vitamin D in the sample. In particular, the method of the present invention distinguishes between cholecalciferol, pre-cholecalciferol and isomers thereof when using a sample of cholecalciferol of about 2800 IU and distinguishes one or more cholecalciferol ester adducts. It is sufficiently sensitive and selective to detect or to detect one or more pre-cholecalciferol ester adducts.

Three forms of potential vitamin D3 degradation products were observed in the stability studies for the pharmaceutical compositions of the present invention described below (including Example 6). In the stability studies described below, the tablet compositions studied were about 91.4 mg of alendronate sodium, about 26.7 mg cholecalciferol granules, about 131.0 mg microcrystalline cellulose, about 62.4 mg anhydrous lactose, about 9.7 mg Croscarmellose sodium, about 0.8 mg colloidal silicon dioxide and about 3.1 mg magnesium stearate.

As shown in FIG. 5, the structure of vitamin D ₃ comprises conjugated trienes, which perform various thermal and photochemical isomerizations. Five vitamin D ₃ isomers have been identified in exemplary pharmaceutical compositions of the invention (in this example, using vitamin D ₃ (cholecalciferol)): pre-vitamin D ₃ , trans-vitamin D ₃ , and 0.78 Three additional isomers of RRT, 0.96RRT and 1.09RRT, which is a measure of the retention time of compounds by high performance liquid chromatography (HPLC) as described below. The structure for these vitamin D ₃ isomers is shown in FIG. 5. Structural conclusions are based on UV, MS and, in some cases, NMR spectroscopy.

Vitamin D and its isomer pre-vitamin D are known to be thermally interconverted by sigma bound 1,7-hydrogen migration. In vivo, pre-vitamin D has been found to be an intermediate precursor to vitamin D, and both groups are identified at equilibrium concentrations at physiological temperatures, although the equilibrium appears to be significantly towards vitamin D. Since both vitamin D and pre-vitamin D are believed to have the same physiological function, the vitamin D assay usually includes the sum of the two groups when reported. This is described, for example, in USP and Ph. Eur. Consistent with the research paper. Useful stability data indicate that none of the other isomers, with ICH qualification thresholds ranging from 24 months to 1.0% by weight, are stored at 25 ° C./60% RH in a suitable package.

The overwhelming degradation product appeared to be a vitamin D ₃ ester formed by transesterification with the vitamin D ₃ heavy chain triglyceride (MCT) of the granules used in the composition of the vitamin D ₃ in the present invention. The structure of these vitamin D ₃ ester adducts is also shown in FIG. 5. The dominant group corresponds to the n-octanoate (C ₈ ) and n-decanoate (C ₁₀ ) esters of vitamin D ₃ . Pre-vitamin D ₃ ester adducts may be produced by reaction of pre-vitamin D ₃ with triglycerides in vitamin D ₃ compounds or by thermal conversion from vitamin D ₃ esters. Among the quantifiable degradation products, only C ₈ and C ₁₀ vitamin D ₃ ester adducts were shown to increase to a significant degree during stability studies.

Useful stability data should be stored in these groups at relative humidity (RH) where the ICH Qualification Threshold should not reach 1.0 wt% at 24 months, but less than about 30 ° C. and less than about 30%, and these are aspects of the compositions and methods of the present invention. It cannot be expected to generate a safety aspect. It is also seen from the study that after 24 months when stored at RH of less than about 30 ° C. and less than about 30%, the total degradation of the composition of the present invention is less than about 5%.

It can also be seen that vitamin D can undergo autooxidation in solid or liquid phase through induction by free radical initiators or spontaneously to form various products, some of which have been identified. Typical properties of (vitamin D ₃ on and, in particular, in this case) of vitamin D degradation in aspects of the present composition, and vitamin D ₃ self-check oxidation instability, wherein the vitamin D ₃ is converted to an oil or amorphous solid, 20 To a temperature of from 40 ° C. In time, HPLC analysis reveals the appearance of many undecomposed degradation products that exhibit significant destruction of vitamin D ₃ and very low ultraviolet absorption. With longer exposure times, these absorptions continue to decrease as further reactions occur.

A more detailed analysis of the autooxidation of vitamin D ₃ is carried out using azo-bis-isobutyronitrile (AIBN), a free radical initiator. In this experiment, AIBN is used to initiate the autooxidation of vitamin D ₃ in solution. The resulting product profile was characterized by HPLC using UV, mass spectrometry (MS) and evaporative light scattering (ELS) detection. From the results, (a) liquid phase autooxidation can also lead to multiple decomposition products, and (b) autooxidation can lead to gradual destruction of UV chromophores, leading to apparent material loss, while ELS detection is significantly more It was found that it provides good mass recovery and (c) the mass spectrometer m / z ratio and in some cases, the observed UV / vis spectra confirm the oxidative properties of these reaction products.

Radiolabeling studies were conducted in an attempt to characterize vitamin D ₃ degradation in granule formulations used in embodiments of the compositions of the present invention comprising about 70 mg of alendronate and about 2,800 IU (70 μg) of vitamin D ₃ . Tritium labeled vitamin D ₃ was used as a tracer of degraded vitamin D ₃ irrespective of changes in UV absorbing properties. Radiolabeled vitamin D ₃ was incorporated into formulations modeled after the vitamin D granules used in the composition embodiments of the present invention and then analyzed for stability. Antioxidant levels in the model formulations were present at reduced levels from antioxidant levels deemed desirable for commercial formulations to ensure degradation occurred in a reasonable time frame. Samples were analyzed after 14 weeks at 40 ° C./75% RH and 70 ° C. using liquid scintillation counting (LSC) and reverse phase high performance liquid chromatography (RP-HPLC) simultaneously with UV and online radiation detection. Approximately 40% loss of vitamin D ₃ is observed for samples stored at 40 ° C./75% RH, whereas low temperature control shows good stability. The results of this analysis are shown in the radiochromatograms for the degraded samples of FIG. 6, which represent a large area of undegraded degradation, none of which appear as major products. These results provide additional evidence that, if not adequately stabilized, vitamin D ₃ oxidatively decomposes into multiple products with reduced UV absorption, and these products are a major source of vitamin D ₃ loss. Based on these stability analyzes, individual autooxidative degradation products are not expected to approach safety-related levels in the purification aspect of the compositions of the present invention.

The invention also includes a method of measuring the pharmacokinetic parameters of a mammal upon administration of a composition of the invention. The pharmacokinetic parameters that can be measured include the area under the total urine volume, urination volume, serum concentration versus time curve of the tablet, such as, for example, a tablet comprising about 70 mg of alendronate and about 2,800 IU of cholecalciferol. (AUC), steady state maximum plasma concentration (C _max ), time of C _max (T _max ), and plasma concentration intermediate apparent half-life (t _{l / 2} ). These measurements confirm that embodiments of the compositions and methods of the present invention produce pharmaceutically effective levels of alendronate and cholecalciferol in the body (the latter recommended daily dose of vitamin D compounds in the compositions and methods of the present invention). As compared to).

In one aspect, the present invention includes a method for measuring cholecalciferol in human serum after administration of a pharmaceutical composition comprising alendronate and cholecalciferol, the method comprising: (1) alendronate and cholecal to a human Administering a composition comprising ciferol, (2) obtaining a plasma sample from a human, (3) extracting cholecalciferol from the plasma sample to form a first solution, and (4) cholecal of the first solution Reaction of ciperol with dienofil to form one or more die-alder adducts of cholecalciferol, and (5) separating the dieel-adder adduct of cholecalciferol using high performance liquid chromatography (HPLC) separation. And (6) detecting the amount of cholecalciferol in the sample using mass spectroscopy. In one embodiment of this method, the dienophile comprises 4-phenyl-1,2,4-triazolin-3,5-dione (P-TADO or PTAD). In addition, the detection step can be performed in positive ionized form using a heated nebulizer probe, adding deuterated internal standard cholecalciferol to each human plasma sample, and deuterated internal standard with sample cholecalciferol. Extracting, reacting, separating and detecting cholecalciferol may be further included. When this method measures 1 ml of plasma, the limit of quantitation (LOQ) of cholecalciferol is less than about 0.5 ng / ml cholecalciferol. One aspect of the invention is a vitamin D / bisphosphonate composition, wherein 120 hours after administration of the composition, the serum concentration plot of the mammal yields one or more of about 296.4 ng.h / ml of cholecalciferol Least-square average AUC _{(0-120 hours)} , where pharmacokinetic parameters were measured without considering baseline cholecalciferol serum concentrations; Least squared (LS) mean AUC _{(0-120 hours)} of about 297.5 ng.h / ml, where the pharmacokinetic parameters are reference variables using baseline pre-dose serum cholecalciferol concentrations as 0 hour Measured in consideration of serum concentrations) and a minimum squared (LS) mean AUC _{(0-120 hours)} of about 143.1 ng.h / ml (where pharmacokinetic parameters were subtracted from baseline cholecalciferol measured for 120 hours). Was measured taking into account baseline cholecalciferol serum concentrations). In another embodiment, the composition comprises bisphosphonate and cholecalciferol, wherein 120 hours after administration of the composition, the serum concentration plot of the mammal yields one or more of the following: normal for 120 hours at about 5.9 ng / ml Least squares (LS) mean for state maximum plasma concentration (C _max ), where pharmacokinetic parameters were measured without considering baseline cholecalciferol serum concentrations; Least-squared (LS) mean of steady-state maximum plasma concentration (C _max ) for 120 hours of about 5.9 ng / ml (wherein the pharmacokinetic parameters are used as pre-administration 0 hour serum cholecalciferol concentrations). Was determined taking into account baseline cholecalciferol serum concentrations) and the mean squared (LS) mean for steady state maximum plasma concentration (C _max ) of about 4.0 ng / ml (where pharmacokinetic parameters were measured for 120 hours). Measurement was performed taking into account baseline cholecalciferol serum concentrations using the subtraction of baseline cholecalciferol).

The present invention also provides a steady-state maximum plasma concentration of cholecalciferol at the arithmetic mean time (T _max ) of C _max development of about 12 hours in a plasma concentration plot of cholecalciferol in mammals 120 hours after administration of the composition. (C _max ) is obtained and the pharmacokinetic parameters include compositions that do not take into account baseline cholecalciferol serum concentrations. In another embodiment, the composition has a plasma concentration median half-life (t _1/2 ) of the composition's cholecalciferol in the mammal in about 23.8 hours, and the pharmacokinetic parameter uses the subtraction of the measured standard cholecalciferol process. It is measured taking into account the standard cholecalciferol serum concentration.

To determine the pharmacokinetic properties of the compositions of the present invention, a sample study was conducted from a randomized two-part, two-period cross-over study of open-labels in 236 healthy non-pregnant women and men ages 18-65 To perform. In this study, the following Examples 7 Pharmacokinetic Parameters of vitamin D ₃ is administered as described in detail in 70㎎ alendronate / 2800IU of vitamin D ₃ for the purification of vitamin D ₃ 2800IU mixing purified, that the variable _{(AUC (0-120 Time)} , C _max , T _max and serum concentration median apparent half-life (t _1/2 )). In addition, urinary secretion of alendronate is FOSAMAX Study in mixed tablets associated with 70 mg tablets once weekly. In summary, (1) the 70 mg Alendronate / 2800IU vitamin D ₃ mixed tablet according to the present invention was found to have bioequivalent to 70 mg Alendronate tablets in terms of Alendronate bioavailability, and (2) 70 mg of of alendronate / 2800IU vitamin D _3, and mix purified 2800IU of vitamin D ₃ in the tablet containing the (alendronate does not contain) the bioavailability of vitamin D ₃ has been found to be similar, 3 of 70㎎ according to the invention alendronate / Vitamin D ₃ blend tablets of 2800 IU have been found to be generally well tolerated. Thus, for example, weekly-once administration with bisphosphonate / vitamin D compounds of the present invention is recommended for the same period of time as weekly-once administration of bisphosphonate / vitamin D compounds, eg : Provides vitamin D ₃ blood levels and / or therapeutic effects comparable to vitamin D blood levels and / or therapeutic effects resulting from 400 IU of vitamin D) per day.

These and other aspects of the invention are again described in the following non-limiting examples.

Example

The following examples further describe and illustrate aspects within the scope of the invention. As many variations of the invention are possible without departing from the spirit and scope of the invention, the examples are provided for illustrative purposes only and are not intended to limit the invention.

Example 1

Bisphosphonates and Vitamin D Tablets

The finished drug product is a combination tablet containing allenetronate sodium (about 70 mg equivalent of free acid anhydride) and vitamin D ₃ (about 2800 I.U. (about 70 μg)), the components of which are identified in Table 1. Excipients are both schematic and were chosen to achieve maximum physical and chemical stability.

The tablets obtained are used according to the methods of the invention, for example for the prevention, inhibition, reduction or treatment of osteoporosis. Similarly, for an alendronic acid active base, tablets are prepared that include other relative weights of alendronate, including about 2.5 mg, 5 mg, 8.75 mg, 17.5 mg, 70 mg, 140 mg, 280 mg, 560 mg or 1120 mg per tablet. However, the present invention is not limited thereto. Similarly, tablets are prepared comprising different relative weights of vitamin D ₃ per unit dose, including but not limited to about 1,400, 2,800, 5,600, 7,000 IU, 8,400 IU, 14,000 IU, 28,000 IU or 36,000 IU per tablet. It doesn't happen. Such tablets may be administered at intervals ranging from weekly to 2 months-1 times.

Example 2

Bisphosphonates and Vitamin D Compositions

Compositions comprising bisphosphonates and vitamin D can be prepared using the mixing and formulation techniques described herein. For the alendronic acid active base, a composition containing about 35 mg of alendronate and about 5,600 IU of vitamin D ₃ can be prepared using the following relative weight components.

The formulations obtained are used according to the methods of the invention, for example to prevent, inhibit, reduce or treat osteoporosis. Similarly, for alendronic acid active bases, formulations containing other relative weights of alendronate are prepared, which include about 2.5 mg, 5 mg, 8.75 mg, 17.5 mg, 70 mg, 140 mg, 280 mg, 560 mg or 1120 mg per tablet. However, the present invention is not limited thereto. Similarly, formulations are prepared comprising different relative weights of vitamin D ₃ per unit dose, including, but not limited to, about 1,400, 2,800, 5,600, 7,000 IU, 8,400 IU, 14,000 IU, 28,000 IU, or 36,000 IU per formulation. It is not limited. Such formulations may be administered at intervals ranging from weekly to 2 months-1 times. These formulations can be, for example, tablets or capsules.

Example 3

Alendronate and Vitamin D Tablets

For Alendronic acid active bases, tablets containing about 70 mg of alendronate, and 2800 IU of vitamin D ₃ are prepared by the methods described herein using the following relative weights of components:

The tablets obtained are used according to the methods of the invention, for example to prevent, inhibit, reduce or treat osteoporosis. Similarly, for an alendronic acid active base, tablets are prepared that include other relative weights of alendronate, including about 2.5 mg, 5 mg, 8.75 mg, 17.5 mg, 70 mg, 140 mg, 280 mg, 560 mg or 1120 mg per tablet. However, the present invention is not limited thereto. Similarly, tablets are prepared comprising different relative weights of vitamin D ₃ per unit dose, including but not limited to about 1,400, 2,800, 5,600, 7,000 IU, 8,400 IU, 14,000 IU, 28,000 IU or 36,000 IU per tablet. It doesn't happen. Such tablets may be administered at intervals ranging from 1 week to 2 months-1 times.

Example 4

Vitamin D for Alendronate Absorption ₃ Effect of (powder form)

Two-term crossover for 14 healthy, 18-85 year old, non-pregnant women and men when administered in a single dose to test the interaction of vitamin D ₃ (cholecalciferol) in powder form with alendronate The study was conducted. The subject was given one Alendronate 70 mg tablet in each period. A single dose of Vitamin D ₃ 5600 IU was co-administered with the Alendronate tablet in one of two periods, based on the computer-generated subject distribution schedule. When vitamin D ₃ was administered with alendronate, the vitamin D ₃ powder was reconstituted in 60 ml of pure tap water and administered to the subject together with the alendronate tablet (treatment A). The bottle of vitamin D ₃ was washed, filled three times with 60 ml of pure tap water, and then administered to each subject. Thus, a total of 240 ml volume of pure tap water was administered with vitamin D ₃ . When Alendronate was administered alone, 240 ml volume of pure tap water was administered with this dose (treatment B). More than 14 days washes were separated in each period. Treatment outlines and allocations are in Table 4.

Subjects were segregated into study units in the evening before each treatment. After fasting overnight (except water), subjects were administered individual treatment. Subjects continued to fast after drug administration until a limited meal was administered 2 hours after administration.

Clinical supply information is in Table 5. Compositions and analytical results for the Alendronate tablets and vitamin D ₃ used in this study are listed in Tables 6 and 7.

All doses were administered after fasting overnight (except water). Subjects received a single Alendronate 70 mg tablet with 240 ml of pure tap water. When subjects were given a dose of 5600 IU of vitamin D ₃ with alendronate, the subject was instructed to re-suspend and co-administered an allandronate 70 mg tablet with vitamin D ₃ doses (reformed in the study and reconstituted in water). . The total volume of liquid administered with each alendronate dose was 240 ml. The subject stayed more fast for 2 hours after study drug administration, and then provided a limited meal. Subjects were kept straight for 2 hours between drug administration and limited meals. Each dose period was separated at least 14 days apart.

Urine samples for the Alendronate assay were collected for pharmacokinetic analysis over the following intervals: -2 to 0 hours before dosing, 0 to 8 hours after dosing, 8 to 24 hours after dosing and 24 to 36 hours after dosing. Urine harvests obtained over a 2-hour period immediately prior to study drug administration were used for baseline alendronate measurements. All urine samples were collected in pre- weighed polypropylene containers. For 0--8-, 8-- 24- and 24--36-hour urine collections after administration, 12.5 g of boric acid was added to the vessel as a preservative at the start of the time interval. At the end of each fixed time collection interval, the total urine harvest weight was measured and specific gravity and pure volume were measured. Urine samples were acidified in situ. 5 ml of 6.0N hydrochloric acid (HCl) was added per 200 ml of urine to bring the pH of the urine specimen to 2.0 or less. After acidification, the urine specimen was stirred and the sample was aliquoted into a polypropylene container and frozen (-20 ° C.) until completion of the high performance liquid chromatography (HPLC) assay. The total urine volume for each period was measured using the total urine volume for each period, including the volume of boric acid and HCl.

Analytical methods for measuring alendronate in human urine include three different manipulations: (1) separation of analyte and internal standard (palmidronate) from urine, (2) production of strong fluorescent derivatives, and (3) obtaining HPLC separation and fluorescence detection of one derivative. Alendronate and internal standards were co-precipitated from urine by the addition of calcium chloride and sodium hydroxide using naturally occurring phosphates. The pellet separated by centrifugation was reconstituted in 1M hydrochloric acid and applied to an anion-exchange diethylamine (DEA) cartridge in acetate-buffer at pH 4. Alendronate was eluted from a DEA cartridge with a solution of 0.20 M sodium citrate and 0.20 M dibasic sodium phosphate (adjusted to pH 9). Alendronate was derived at room temperature in the presence of N-acetyl-D-penicillamine using 2,3-naphthalenedicarboxyaldehyde. The derivative was then applied to a non-silica polymer column consisting of a copolymer of styrene and divinyl benzene. The mobile phase initially consists of 85% 0.025M sodium citrate, 0.025M dibasic sodium phosphate (pH 6.95) and 15% acetonitrile at a flow rate of 1 ml / min. The endogenous components of the eluted urine were then removed by increasing the acetonitrile to 50%. In human urine, assays were validated with a coefficient of change of up to 10% between 5 ng / ml and 125 ng / ml. 5 ml urine samples were needed to obtain a detection limit of 1 ng / ml.

Aliquots analyzed by the total volume of urine (including boric acid and hydrochloric acid) for the interval at which the total urine volume of the alendronate for a given interval (-2 to 0, 0 to 8, 8 to 24, 24 to 36 hours) It was measured by doubling the concentration of allendronate in water.

Total urination for 70 mg Alendronate tablets + 5600 IU Vitamin D ₃ and 70 mg Alendronate tablets alone was compared using analysis of a variable (ANOVA) model suitable for a two-term crossover design. The ANOVA model contained factors for order, subject (order), duration and treatment. Total urination volume was modified long. The results from the Shapiro-Wilk test on the normal did not suggest a departure from the assumption of the ANOVA model with a plot of the residual amount from the model. To assess the relative bioavailability of the 70 mg Alendronate tablet plus Vitamin D ₃ vs. 70 mg Alendronate tablet alone, 95% CI based on the t-distribution was estimated for GMR for total urination. In addition, the likelihood was also calculated after the de facto GMR was above the clinically important boundary of 0.50.

One subject was dropped from the assay because this particular subject had a urinary alendronate concentration below the quantification limit for both treatments in all three collection intervals (0-8, 8-24 and 24-36 hours). . Due to some imbalance in the indication of the order of treatment, the least square method for total urination is reported. Data from the subject was excluded from the analysis.

Least squares method was obtained by inverse transformation from the ANOVA model. All p values were completed with three decimal places before reporting. Results reported with p ≦ 0.050 are considered to be statistically significant.

Table 8 shows the total urine output of the alendronate as 70 mg allenronate tablets plus vitamin D ₃ and 70 mg allenronate tablets for each subject. For total urination volume of alendronate, together with GMR, the basic statistics with the corresponding 95% CI are in Table 9.

The least squares geometrical method for total urination was 183.61 for 70 mg Alendronate tablets + 5600 IU Vitamin D ₃ and 157.97 μg for 70 mg Alendronate tablets alone. The GMR and corresponding 95% CI for 70 mg Alendronate + Vitamin D ₃ compared to Alendronate alone was 1.16 (0.74, 1.83). The later possibility that the GMR may be above the clinically important boundary of 0.50 is 0.999.

Example 5

Vitamin D (in Alendronate / Vitamin D Tablets) for Alendronate Absorption ₃ Effect

To test the efficacy of the interaction between alendronate and orally administered vitamin D ₃ , 14 healthy adult subjects (6 males and 8 females, aged 33-61) received a single 70 mg tablet of alendronate, a vitamin D _In the absence of ₃ and with a powder dose of vitamin D ₃ (5600 IU) suspended in 240 ml of water. The study was designed to be open and randomly cross two-way. The purpose of the study is compared with the administration of alendronate in the absence of vitamin D _3, it was to obtain a preliminary assessment of the relative bioavailability of alendronate after administration of 70mg tablets with the Vitamin D _3.

Alendronate was administered orally as 70 mg tablets in each of the 2-periods. In one period, the tablets were administered with reconstituted vitamin D ₃ in pure tap water, and in another period the tablets were dissolved in pure tap water alone. Urine was collected 2 hours before and 36 hours after each alendronate administration for analytical determination of secreted alendronate. Relative bioavailability was assessed based on the total urine recovery of Alendronate over 36 hours post dose.

The urine recovery of alendronate after administration of 70 mg alendronate in the absence of vitamin D ₃ was 202 μg with 90% CI of (126 μg, 279 μg), and when recovered following the 70 mg dose administered with vitamin D ₃ , (159 μg , 316 μg) with an average of 238 μg. The geometric mean ratio (90% CI) was estimated to be 1.18 (0.80, 1.74). This investigation indicates that oral administration of vitamin D ₃ with oral administration of alendronate does not have any minimal effect on the bioavailability of alendronate.

Example 6

Vitamin D ₃  And stability studies of alendronate

The stability of the compositions of the present invention in the form of combination tablets containing alendronate sodium (70 mg free acid equivalent) and vitamin D ₃ (2800 IU / 70 μg) was studied. Table 10 contains the tablet compositions of the Alendronate / Vitamin D combined tablet embodiments. All excipients are schematic and were chosen to achieve maximum physical and chemical stability.

Alendronate assays and dissolution methods are FOSAMAX Pre-column 9-fluorenylmethyl chloroformate (FMOC) derivatization can be used with reversed phase HPLC, similar to the methods already reported for purification. The vitamin D ₃ and degradation products method may also be a reverse phase, gradient HPLC method (RP-HPLC) capable of dissociating and quantifying a number of effective degradation products of vitamin D ₃ and vitamin D ₃ . Content uniformity and dissolution assays of vitamin D ₃ may also use reverse phase HPLC. The dissolution method may use surfactant medium (1% SDS) due to the poor aqueous stability of vitamin D ₃ . Due to the low vitamin D ₃ potency (70 μg) of the combined tablets, three tablets in 500 ml of medium can be used to obtain a suitable signal.

52-week assay and degradation data are provided below for batches of combination tablets stored at 30 ° C./65% RH and 40 ° C./75% RH (see Tables 11 and 12). These data show the acceptable stability of the composition embodiments of the present invention, but the data generated actually indicate that there is a mild degradation of vitamin D ₃ . The higher the temperature, the more decomposition was found in the aluminum foam and HDPE bottles in the absence of desiccant.

Example 7

Vitamin D ₃ And Pharmacokinetics of Alendronate

Open marker, randomized, two-part, two-period, crossover studies were performed on 236 healthy, 18-65 year old, non-pregnant women and men. The study was conducted in two parts (Parts I and II), each consisting of two periods of crossover design. Each subject participated in only one part of the study (ie, each subject participated only in Part I or II). Subjects entered the study sequentially into each part of the study, with washout periods of 12 days or more between treatment periods within each part of the study. Part I included Treatments A and B and Part II included Treatments A and C. Treatment consisted of: Treatment A—Single dose of 70 mg Alendronate / 2800IU Vitamin D ₃ combination tablet, according to Table 7-3 below; Treatment B—Single dose of 70 mg Alendronate tablet, according to Table 7-2 below; Treatment C—Single dose of 2800 IU Vitamin D ₃ tablets (containing placebo excipients replacing alendronate) according to Tables 7-4 below.

In Part I, urine was collected over 2 hours prior to dose administration and 36 hours after dosing in each period to determine the total urinary volume of alendronate. In Part II, for serum vitamin D ₃ measurements at selected times over -24, -18, -12 and -6 hours prior to dosing at each period, at 0 hours (immediately prior to drug administration), and 120 hours after dose administration Blood samples were collected.

Part I of the study evaluated the bioequivalents of Alendronate in 70 mg Alendronate / 2800IU Vitamin D ₃ combination tablets according to Table 16 below and 70 mg Alendronate tablets according to Table 15 below. Part II of the study evaluated the serum pharmacokinetics (AUC _{0-120 hours} , C _max ) of vitamin D ₃ obtained after administration of 70 mg Alendronate / 2800IU Vitamin D ₃ combination tablet and 2800IU Vitamin D ₃ tablet. The 2800 IU Vitamin D ₃ tablet contained 2800 IU Vitamin D ₃ and an inert excipient in the Alendronate / Vitamin D ₃ combination tablet.

The primary pharmacokinetic parameter in Part I was the total urination volume of alendronate 0-36 hours after oral dose administration. Determination of the total urination volume of alendronate is consistent with previous studies characterizing the oral bioavailability of alendronate via urination, as the plasma concentrations are low and difficult to detect after oral administration.

The primary pharmacokinetic parameters in Part II were AUC _{0-120 hours} and C _max of vitamin D ₃ . Blood was collected to determine serum vitamin D ₃ concentrations at the following time points in each period: -24, -18, -12, -6 hours before dosing, 0 hours (immediately before drug administration), and dosing 2, 3, 5, After 7, 16, 24, 36, 48, 72, 96 and 120 hours.

In the study medication before adjustment by the return of serum vitamin D ₃ concentration for 24 hours over 24 hours in the environment of vitamin D ₃ It served as an indicator of the behavior of endogenous levels. Since vitamin D ₃ is synthesized through skin-to-ultraviolet exposure, subjects live in study units and are not exposed to direct sunlight during the pharmacokinetic-sampling period (e.g., for 144 hours from 24 hours before to 120 hours after administration). ). Subjects need to apply sunscreen (SPF 45) and limit sun exposure throughout the study, including the washout period. The subject may also have foods known to be high in vitamin D ₃ (eg salmon, herring, mackerel, cod, tuna, swordfish oysters and sardines). In addition, eating foods known to supplement vitamin D ₃ (eg, certain grains, fortified milk, and certain yogurts) have been restricted.

In Part I each subject received a single oral dose of 70 mg Alendronate / 2800 IU vitamin D ₃ combination tablet and a single oral dose of 70 mg Alendronate in a randomized, crossover fashion. In Part II, subjects received a single oral dose of 70 mg Alendronate / 2800 IU vitamin D ₃ combination tablet and a single oral dose of 2800 IU vitamin D ₃ tablet in a randomized crossover fashion. Doses were administered with 240 ml of pure tap water starting overnight at 2100 hours prior to dosing and fasting overnight (except water). The subject was instructed to stay upright (sit or stand at least 45 °) between the medication and the restricted meal. Subjects fasted until a standard meal administered at a dose after 2 hours. The method for administering the Alendronate / Vitamin D ₃ combination tablet is the same as for Alendronate.

The compositions administered in the study are described in Tables 14-17 below.

The area under the serum concentration versus time curve (AUC _{0-120 hours} ) after _{0-120 hours} of administration was calculated using the unregulated concentration of vitamin D ₃ (C _t ) by the trapezoidal method for final sample collection. Samples with concentrations below the limit of assay (LOQ) of quantification were assigned a value of zero for calculation. The maximum concentration observed (C _max ) and the time of C _max (T _max ) were obtained by examining the measured concentration of vitamin D ₃ in serum and the actual recording time of sample collection. The concentration profile of vitamin D _{3 in} the serum was also measured in three different ways, two of which considered the basal vitamin D ₃ serum concentration in the manner discussed below. AUC _{0-120 hours} , C _max and T _max were calculated in the same manner.

The bioavailability of the 70 mg Alendronate tablets / 2800 IU Vitamin D ₃ combination tablets compared to the 70 mg Alendronate alone tablets was evaluated using GMR for the total urination volume of the Alendronate from the Alendronate / Vitamin D ₃ combination tablets versus the 70 mg Alendronate alone tablets. Relative bioavailability of 70 mg Alendronate / 2800IU Vitamin D ₃ combination tablets for 2800 IU Vitamin D ₃ tablets alone was assessed using GMR (Alendronate + Vitamin D ₃ / Vitamin D ₃ alone) for AUC _{0-120 hours} and C _max It was.

After administration of the 70 mg Alendronate / 2800IU Vitamin D ₃ combination tablet and the 2800IU Vitamin D ₃ tablet, vitamin D ₃ single dose pharmacokinetics were compared using three different approaches. In a first approach, vitamin D ₃ pharmacokinetics to endogenous vitamin D ₃ serum concentrations were compared after administration of the two treatments.

Using this approach, the vitamin D ₃ single dose pharmacokinetics to endogenous vitamin D ₃ serum concentrations after administration of the 70 mg Alendronate / 2800 IU Vitamin D ₃ combination tablet and the 2800 IU Vitamin D ₃ tablet were developed using a two-term, crossover design suitable ANOVA model. Compared using. Suitable strains were used for pharmacokinetic parameters (ie, long strains for AUC _{0-120 hours} and C _max were graded T _max and reversed for apparent t _1/2 ). Inverse strain base statistics and estimation results were reported. Assumptions of the ANOVA model were tested for normal. Normal assumptions were generally met for AUC _{0-120 hours} and C _max .

Alendronate 70mg / 2800IU compared to vitamin D ₃ from vitamin D ₃ tablets 2800IU tablet formulation in order to assess the bioavailability of vitamin D _3, 90% CI of the t- distribution with underlying AUC _0-120 of _time and C _max GMR (70 mg Alendronate / 2800 IU Vitamin D ₃ Combination Tablets / 2800 IU Vitamin D ₃ Tablets) were then calculated and compared to the predefined bioequivalent boundaries of (0.80, 1.25). The comparison between basal statistics and treatment also provided the T _max of vitamin D ₃ .

As another method of measuring pharmacokinetic parameters, the importance of vitamin D ₃ prior to administration in plasma has led to the development of a model for changes observed in vitamin D ₃ concentrations, particularly during the experimental period. The model allows subtraction of the distribution from baseline vitamin D ₃ serum concentrations and allows the evaluation of pharmacokinetic parameters that arise exclusively from oral administration of this compound. The model is based on the following assumptions: (1) If no exogenous vitamin D ₃ is administered, the background concentration changes as a function of time in a substantially linear fashion (C _t = C _i + C _m · t, where C _i And C _m is the intercept and slope of the straight line and t is the number of times proportional to dose administration); (2) The endogenous and pharmacokinetic behavior of ingested vitamin D ₃ is independent of one another, ie the body is endogenous and useful in the same way, with or without additional doses of vitamin D ₃ (therapeutic and non-dose treatments in this study). ) And consumed vitamin D ₃ similarly in the presence of varying amounts of the compounds of the invention in the body prior to administration of the dose; (3) Return of the concentration profile to baseline after administration occurs in a pharmacokinetic method similar to that observed when exogenous compounds (most drug products) are administered (ie, the return phase to baseline will be a linear log).

Based on these assumptions, the function describing the basis of the form C _t = C _i + C _m t and the sum of the two-compartment models (see equation below) is based on the Microsoft Solver Routine (EXCEL) method by the least-squares minimization method. The generalized reduction gradient nonlinear optimization method performed with was used to fit the individual C _t to t profiles (-24 to 120 hours after administration). Suitably it was suppressed to produce an end phase of the approach to the baseline approaching linear log behavior. Next, the best fit factor was used to interpolate baseline values in the range of 0 to 120 hours after administration, and the interpolated baseline was subtracted from each profile.

In the above formula,

t is the time for dose administration,

C _t is the expected concentration of vitamin D _{3 in} the serum,

C _i is the expected value of the baseline at t = 0,

C _m is the slope of the expected baseline,

A, B, k _d , k _el and k _a are parameters of a two compartment model with primary absorbance and t _lag is the individual delay in absorbance after oral administration of vitamin D ₃ .

The pharmacokinetic parameters (AUC _{0-120 h} , C _max , T _max ) measured using this method were calculated in the same way as described using the first measurement method. Basic statistics on the total urination volume of alendronate over 36 hours are shown in Table 18 below. After a single dose, the LS mean for the total urination volume of the alendronate was 197.5 and 191.9 μg for the 70 mg alendronate / 2800IU vitamin D ₃ combined tablet and the 70 mg allendronate alone tablet, respectively. The GMR and corresponding 90% CI for the total urine volume of the alendronate (70 mg Alendronate / 2800 IU Vitamin D ₃ combined tablets versus 70 mg Alendronate alone tablet) was 1.03 (0.91, 1.17). The 90% CI corresponded to the predefined bioequivalence boundaries of (0.80, 1.25).

For plasma measurements, the LS mean for vitamin D ₃ AUC _{0-120 hours} (without taking into account endogenous vitamin D ₃ serum concentrations) was 70 mg Alendronate / 2800 IU vitamin D ₃ combination tablets and The 29800 and 337.9 ng h / ml for 2800 IU vitamin D ₃ tablets, respectively (Table 19). AUC _{0-120 hours} GMR (Alendronate + Vitamin D ₃ Combination Tablets / Vitamin D ₃ Tablets) was 0.88 and 90% CI was (0.81, 0.95).

Without considering endogenous vitamin D ₃ serum concentrations, the LS mean for vitamin D ₃ AUC _{0-120 hours} was 70 mg Alendronate / 2800 IU vitamin D ₃ combination tablets and 5.9 and 6.6 ng / ml for 2800 IU Vitamin D ₃ tablets, respectively (Table 20). For C _max GMR (Alendronate + Vitamin D ₃ Combination Tablets / Vitamin D ₃ Tablets) was 0.89 and 90% CI was (0.84, 0.95). AUC _{0-120 hours} and C _max GMR without taking vitamin D ₃ serum concentrations corresponded to the predefined bioequivalence limits of (0.80, 1.25).

Table 21 shows the results of the statistical analysis for vitamin D ₃ T _max , obtained from the serum profile without considering endogenous vitamin D ₃ serum concentrations. The median T _max for vitamin D ₃ in the presence and absence of alendronate was 12.0 and 9.0 hours, respectively. No significance between treatment differences was observed (p value> 0.200).

LS mean for vitamin D ₃ AUC _{0-120 hours} is 70 mg Alendronate / 2800 IU vitamin D ₃ combination tablets and The 29800 and 336.7 ng h / ml for 2800 IU vitamin D ₃ tablets, respectively (Table 22). AUC _{0-120 hours} GMR (Alendronate + Vitamin D ₃ Combination Tablets / Vitamin D ₃ Tablets) was 0.88 and 90% CI was (0.82, 0.95).

LS mean C _max of vitamin D ₃ is shown in Table 23 below, 70 mg Alendronate / 2800 IU vitamin D ₃ combination tablets and 5.9 and 6.6 ng / ml for 2800 IU Vitamin D ₃ tablets, respectively. C _max GMR (Alendronate + Vitamin D ₃ Combination Tablets / Vitamin D ₃ Tablets) was 0.90 and 90% CI was (0.85, 0.95). AUC _{0-120 hours} using pre-dose vitamin D ₃ concentration and 90% CI for C _max GMR as time variables corresponded to the predefined bioequivalence limit of (0.80, 1.25).

Model of the basic baseline vitamin D ₃ concentration of the vitamin D ₃ was measured using a The results of the data analysis for AUC _{0-120 hours} are summarized in Table 24. The LS mean for vitamin D ₃ AUC _{0-120 hours} , measured using model-base vitamin D ₃ baseline concentrations, was 70 mg Alendronate / 2800 IU vitamin D ₃ combined tablets and 143.1 and 169.1 ng h / ml for 2800 IU vitamin D ₃ tablets, respectively. AUC _{0-120 hours} GMR (Alendronate + Vitamin D ₃ Combination Tablets / Vitamin D ₃ Tablets) was 0.85 and 90% CI was (0.76, 0.94). The lower 90% CI falls directly below the predefined lower limit of 0.80.

The LS mean C _max of vitamin D ₃ , measured using model-base vitamin D ₃ baseline concentrations, is shown in Table 25, with 70 mg Alendronate / 2800IU vitamin D ₃ combined tablets and 4.0 and 4.6 ng / ml for 2800 IU Vitamin D ₃ tablets, respectively. C _max GMR (Alendronate + Vitamin D ₃ Combination Tablets / Vitamin D ₃ Tablets) was 0.88 and 90% CI was (0.83, 0.93). The 90% CI for C _max GMR measured using model-base vitamin D ₃ baseline concentrations corresponded to the predefined bioequivalence limit of (0.80, 1.25).

Table 26 shows the results of the statistical analysis for vitamin D ₃ T _max , obtained from the profile measured using the model-based baseline vitamin D ₃ concentration. The median T _max for vitamin D ₃ in the presence and absence of alendronate was 12.0 and 9.0 hours, respectively. No significance was observed between treatment differences (p value> 0.200).

The data analysis of t _1/2 of vitamin D ₃ , measured using the model-base vitamin D ₃ background concentration, is summarized in Table 27. The harmonic mean apparent t _1/2 for vitamin D ₃ in the presence and absence of alendronate was 24.0 and 25.5, respectively. No significant difference was observed between treatment differences (p value> 0.200).

Example 8

Decomposition detection method

A method for the combination assay of Alendronate / Vitamin D ₃ combination tablets (70 mg Alendronate / 2800 IU Vitamin D ₃ ) has been developed. Vitamin D ₃ is extracted from 15 tablets in about 50 ml of 5% water / 95% methanol diluent. The solution is stirred for 10 minutes, sonicated for 30 minutes and stirred for another 3 hours. Samples are centrifuged and 100 μL of supernatant is injected into a Phenomenex Phenosphere 80 μs ODS (1) column (150 × 4.6 mm, 3 μm) for reverse phase HPLC analysis. The method is a 65 minute gradient method using a detection wavelength of 265 nm. The vitamin D ₃ precursor and the vitamin D ₃ peak are both quantified and combined to calculate the total amount of vitamin D _{3 in} the sample. This method is effective for satisfactory specificity, linearity, recovery, precision, reproducibility, solution stability, sensitivity and stiffness.

Exemplary chromatography conditions are described below:

Excipient peaks and degradation with representative retention time and relative retention time (RRT) for vitamin D ₃ are shown in Table 28. The major degradation pathways of vitamin D ₃ are photoisomerization, thermoisomerization and transesterification, as shown in FIG. 5.

The limit of quantification (LOQ) was determined by injecting different concentrations of vitamin D ₃ solution and selecting the lowest concentration with a signal-to-noise ratio of 10 or greater. The LOQ is measured as about 9 ng / ml (injection capacity 100 μL), which is 0.04% of the method concentration with an average signal to noise ratio of 11.1 for 10 replicate measurements.

Example 9

Once-weekly diet

Alendronate and Vitamin D tablets, or other solid dose formulations containing about 70 mg of Alendronate, and about 5,600 IU of vitamin D, can be prepared for the Alendroic acid active base (see Examples 1, 2, and 3). Tablets or other solid dosage forms may be administered orally to a subject once a week, preferably about once every seven days (eg, every Sunday) for a period of at least one year. Such a method of administration is expected to be useful and convenient to provide vitamin D nutrition while treating or preventing osteoporosis. This method is also expected to improve the acceptance and acceptance of subjects and to be useful for ensuring that all subjects taking bisphosphonates receive adequate vitamin D nutrition.

Alternatively, other solid dose formulations containing about 70 mg of alendronate and about 2,800 IU of vitamin D may be prepared for the alendronate and vitamin D tablets or allandroic acid active bases (eg, Example 1 and See 3). Tablets or other solid dosage forms may be administered orally to a subject once a week, preferably about once every seven days (eg, every Sunday) for a period of at least one year. Such a method of administration is expected to be useful and convenient to provide vitamin D nutrition while treating or preventing osteoporosis. This method is also expected to improve the acceptance and acceptance of subjects and to be useful for ensuring that all subjects taking bisphosphonates receive adequate vitamin D nutrition.

Alternatively, other solid dose formulations containing about 35 mg to about 70 mg of alendronate, and about 2,800 IU of vitamin D, may be prepared for the alendronate and vitamin D tablets or allandronic active bases (eg, Examples See 3). Tablets or other solid dosage forms may be administered orally to a subject once a week, preferably about once every seven days (eg, every Sunday) for a period of at least one year. Such a method of administration is expected to be useful and convenient to provide vitamin D nutrition while treating or preventing osteoporosis. This method is also expected to improve the acceptance and acceptance of subjects and to be useful for ensuring that all subjects taking bisphosphonates receive adequate vitamin D nutrition.

For Alendronate and Vitamin D tablets or Alendroic acid active bases, other solid dose formulations containing about 280 mg of Alendronate and about 5,600 IU of Vitamin D can be prepared (see, eg, Examples 2 and 3). . Tablets or other solid dose formulations may be administered orally to a subject once a week, preferably about once every seven days (eg, every Sunday) for a period of at least 1 to 6 months. This method of administration is expected to be useful and convenient to provide vitamin D nutrition while treating Paget's disease. This method is also expected to improve the acceptance and acceptance of subjects and to be useful for ensuring that all subjects taking bisphosphonates receive adequate vitamin D nutrition.

Alternatively, other solid dose formulations containing about 280 mg of alendronate and about 2,800 IU of vitamin D can be prepared for the alendronate and vitamin D tablets or allandronic active bases (see, eg, Example 3). . Tablets or other solid dose formulations may be administered orally to a subject once a week, preferably about once every seven days (eg, every Sunday) for a period of at least 1 to 6 months. This method of administration is expected to be useful and convenient to provide vitamin D nutrition while treating Paguet's disease. This method is also expected to improve the acceptance and acceptance of subjects and to be useful for ensuring that all subjects taking bisphosphonates receive adequate vitamin D nutrition.

Alendronate and Vitamin D For tablets or allandronic active bases, other solid dose formulations containing about 280 mg of alendronate and about 5,600 IU or 2800 IU of vitamin D can be prepared (eg, Examples 1, 2). And 3). Tablets or other solid dose formulations may be administered orally to a subject once a week, preferably about once every seven days (eg, every Sunday). This method of administration is expected to be useful and convenient to provide vitamin D nutrition while treating metastatic bone disease. This method is also expected to improve the acceptance and acceptance of subjects and to be useful for ensuring that all subjects taking bisphosphonates receive adequate vitamin D nutrition.

It will be apparent to those skilled in the art that various modifications can be made in the present invention, which are methods and compositions for inhibiting bone absorption, without departing from the scope and spirit of the invention or claims. In addition, the present invention and the appended claims are intended to include modifications, variations, and equivalents of the methods and compositions for inhibiting bone absorption of the present invention.

Compositions and methods are described for preventing, inhibiting, reducing and treating conditions and diseases associated with abnormal bone absorption in mammals, such as osteoporosis. Embodiments of the compositions of the present invention include a pharmaceutically effective amount of alendronate and vitamin D ₃ suitable for weekly to weekly administration. The compositions and methods of the present invention provide vitamin D nutrition during bisphosphonate treatment to facilitate normal bone formation and mineralization, while minimizing the induction of potential for complications associated with vitamin D deficiency (eg, hypocalcemia and osteomalacia).

1 illustrates the metabolism of vitamin D ₃ .

2 shows the chemical formulas of cholecalciferol and alendronate monosodium trihydrate.

3 shows the chemical formulas of vitamin D ₂ and vitamin D ₃ .

4 shows a schematic summarizing an embodiment of a method of making a composition of the present invention.

5 depicts the thermal and photochemical isomerization and transesterification of vitamin D ₃ .

6 shows the results of radiolabeled studies of vitamin D ₃ degradation.

Claims (26)

A pharmaceutical composition comprising bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates of the bisphosphonates, mixtures thereof and vitamin D compounds. The pharmaceutical composition of claim 1, wherein the bisphosphonate is represented by the following formula (1). Formula 1 In the above formula, R ₁ is independently selected from the group consisting of H, OH and Cl, R ₂ is independently CH ₃ , Cl, CH ₂ CH ₂ NH ₂ , (CH ₂ ) ₃ NH ₂ , CH _2-3 -pyridyl, CH ₂ -S-phenyl-Cl, CH ₂ CH ₂ N (CH ₃₎ (cyclopentyl), CH ₂ - imidazole, CH ₂ -2- imidazo-pyridinyl, N- _{(cycloheptyl), CH 2 CH (CH 3} ) 2, (CH 2) 5 NH 2 and CH ₂ - 1-pyrrolidinyl and mixtures thereof. The pharmaceutical composition of claim 1, wherein the bisphosphonate comprises alendronate or a pharmaceutically acceptable salt thereof. 4. The pharmaceutical composition of claim 3, wherein the pharmaceutically acceptable salt of the alendronate is selected from the group consisting of alendronate monosodium, alendronate monosodium monohydrate, and alendronate monosodium trihydrate. The pharmaceutical composition of claim 1, comprising about 100 to about 36,000 IU of vitamin D compound. The pharmaceutical composition of claim 1 comprising from about 0.5 to about 1120 mg of bisphosphonates or pharmaceutically acceptable salts, derivatives or hydrates of the bisphosphonates, or mixtures thereof. The pharmaceutical composition of claim 1 comprising about 2,800 IU of cholecalciferol and about 70 mg of alendronate or a pharmaceutically acceptable salt, derivative, or hydrate thereof, or a mixture thereof. The pharmaceutical composition of claim 1 comprising about 5,600 IU of cholecalciferol and about 70 mg of alendronate or aldehyde acceptable pharmaceutically acceptable salts, derivatives or hydrates thereof, or mixtures thereof. The pharmaceutical composition of claim 1, further comprising one or more excipients selected from the group consisting of fillers, diluents, binders, lubricants, glidants, and disintegrants. The pharmaceutical composition of claim 1, further comprising one or more excipients selected from the group consisting of anhydrous lactose, microcrystalline cellulose, colloidal silicon dioxide, croscarmellose sodium and magnesium stearate. The method of claim 10, wherein about 0.5 to about 90 weight percent sodium alendronate sodium, about 1 to about 70 weight percent cholecalciferol granules (equivalent to about 0.0005 to about 20 weight percent cholecalciferol), about 10 to about About 80% by weight of anhydrous lactose, about 5 to about 50% by weight of microcrystalline cellulose, about 0.1 to about 5% by weight of colloidal silicon dioxide, about 0.5 to about 10% by weight of croscarmellose sodium and about 0.5 to A pharmaceutical composition comprising about 5% magnesium stearate. 8. The pharmaceutical composition of claim 7, wherein the cholecalciferol comprises pharmaceutical grade cholecalciferol. The pharmaceutical composition according to claim 12, which is suitable for administration at weekly, two-week, one-month, one-month-two and two month intervals. Preparing a powder mixture comprising the alendronate, densifying the powder mixture to form the alendronate mixture, pulverizing and mixing the alendronate mixture and cholecalciferol granules to form the final mixture, and smoothing and compressing the final mixture. A method for producing an alendronate-cholecalciferol composition.  Alendronate, colloidal silicon dioxide, anhydrous lactose, microcrystalline cellulose and croscarmellose sodium are mixed to form a preliminary mixture, the premix and magnesium stearate are mixed to form a first lubricious mixture, and the first lubricious mixture is formed. The roller is densified to form a densified ribbon, the densified ribbon is pulverized to form a lubricious mixture, the lubricious mixture and cholecalciferol granules are mixed to form a second lubricious mixture, and the second lubricious mixture is then converted into a solid A method of making an alendronate-cholecalciferol solid dosage form, comprising the step of compacting. A pharmaceutical composition prepared by the method of claim 15. The method of claim 1, wherein about 0.5 to about 90 weight percent of alendronate sodium, about 0.1 to about 5 weight percent colloidal silicon dioxide, about 10 to about 80 weight percent anhydrous lactose, about 5 to about 50 weight percent Ingredients comprising microcrystalline cellulose, about 0.5 to about 10 weight percent croscarmellose sodium and about 0.5 to about 5 weight percent magnesium stearate are mixed to form a first mixture, and the first mixture is compacted by roller compaction. Formed ribbons, milled densified ribbons to form a lubricating mixture, and about 1 to about 70 weight percent of cholecalciferol granules (equivalent to about 0.0005 to about 20 weight percent of cholecalciferol) A pharmaceutical composition according to claim 1 prepared by a method comprising mixing to form a second mixture and then pressing the second mixture into a solid dosage form. The pharmaceutical composition of claim 7 formulated to contain less than about 1% by weight of each isomer of cholecalciferol after storage for 24 months at less than about 30 ° C. and less than about 30% relative humidity. The pharmaceutical composition of claim 7 formulated to comprise less than about 5% by weight of a degradation product of cholecalciferol after storage for 24 months at a relative humidity of less than about 30 ° C. and less than about 30%. Bisphosphonates, pharmaceutically acceptable salts, derivatives or hydrates of the bisphosphonates, or mixtures thereof and cholecalciferol, wherein the therapeutic effect of cholecalciferol is about 400 IU per day when administered for 1 week A pharmaceutical composition suitable for once-weekly administration, substantially similar to the therapeutic effect of cholecalciferol of. 2. The plasma concentration plot of claim 1 wherein the bisphosphonate is an alendronate sodium and the plasma concentration plot after administration of the alendronate sodium of the composition to the mammal is a plasma concentration plot after administration of 70 mg of alendronate sodium to the mammal in the absence of cholecalciferol. Substantially similar pharmaceutical compositions. 2. The plasma concentration plot of claim 1 wherein the bisphosphonate is sodium alendronate and the plasma concentration plot after administration of the cholecalciferol of the composition to the mammal comprises a plasma concentration plot after administration of 2800 IU cholecalciferol to the mammal in the absence of allendronate. Substantially similar pharmaceutical compositions. The mammalian serum concentration plot of claim 1, wherein the serum concentration plot of the mammal is 120 min after the least squares (LS) mean AUC _{(0-120 h)} of cholecalciferol of about 296.4 ng.h / ml. , Pharmacokinetic parameters were measured without considering baseline cholecalciferol serum concentrations); Least squared (LS) mean AUC _{(0-120 hours)} of about 297.5 ng.h / ml (where pharmacokinetic parameters are the cohort and baseline cholecalciferol using serum cholecalciferol concentration 0 hours after administration). Measured in consideration of serum concentrations) and a minimum squared (LS) mean AUC _{(0-120 hours)} of about 143.1 ng.h / ml (where pharmacokinetic parameters were subtracted from baseline cholecalciferol measured for 120 hours). Pharmaceutical composition to provide one or more of the following; The plasma concentration plot of the mammal of claim 1, wherein the plasma concentration plot of the mammal is 120 min after the least squares (LS) mean of the steady state maximum plasma concentration (C _max ) for 120 hours of about 5.9 ng / ml. , Pharmacokinetic parameters were measured without considering baseline cholecalciferol serum concentrations); Least-squared (LS) mean of steady-state maximum plasma concentration (C _max ) for 120 hours of about 5.9 ng / ml (wherein the pharmacokinetic parameters are used as pre-administration 0 hour serum cholecalciferol concentrations). Was determined taking into account baseline cholecalciferol serum concentrations) and the mean squared (LS) mean for steady state maximum plasma concentration (C _max ) of about 4.0 ng / ml (where pharmacokinetic parameters were measured for 120 hours). Pharmaceutical composition providing one or more of the following; The steady state maximum of cholecalciferol at the arithmetic mean time (T _max ) of C _max development of about 12 hours in a plasma concentration plot of cholecalciferol in mammals 120 hours after administration of the composition. Pharmaceutical compositions wherein plasma concentrations (C _max ) are obtained and pharmacokinetic parameters are measured without considering baseline cholecalciferol serum concentrations. The method of claim 1, wherein the mammal has a median half-life (t _1/2 ) of the plasma concentration of cholecalciferol in the composition of about 23.8 hours, and the pharmacokinetic parameter is determined using a subtracted measure of the reference cholecalciferol process. A pharmaceutical composition measured in consideration of the reference cholecalciferol serum concentration.