TW200538136A - Compositions and methods for inhibiting bone resorption - Google Patents

Abstract

Disclosed are compositions and methods for preventing, inhibiting, reducing and treating conditions and diseases associated with abnormal bone resorption in mammals, including for example osteoporosis. Embodiments of compositions of the invention comprise a pharmaceutically effective amount of alendronate and vitamin D3 suitable for once-weekly dosing. Compositions and methods of the invention provide vitamin D nutrition during bisphosphonate treatment to facilitate normal bone formation and mineralization while minimizing the occurrence of potential for or the complications associated with vitamin D insufficiency, such as hypocalcaenua and osteomalacia. Also disclosed are methods for manufacturing compositions of the present invention, for measuring stability and degradation of those compositions, and for measuring blood plasma levels of vitamin D.

Description

200538136 IX. Description of the invention: [Technical field to which the invention belongs] The present invention relates to a mouthpiece comprising a bisphosphate compound and a vitamin 0 compound. The invention also relates to methods of using such compositions in, for example, treating, reducing, inhibiting or preventing abnormal bone loss in mammals. The invention further relates to a method for manufacturing a bisphosphate and vitamin!) Composition. [Prior art] A variety of human and other mammalian disorders involve or may be associated with abnormal bone loss. The most common of these conditions is osteoporosis, which is a systemic skeletal disease, characterized by low bone mass and microstructural damage to bone tissue; therefore, bones become more vulnerable and easily fractured. Osteoporosis has caused a worldwide epidemic, and its incidence has increased due to a worldwide extension of lifespan. The major cell type responsible for bone wasting is a multinucleated cell called an osteoclast. Bisphosphate is a known selective inhibitor of osteoclast bone loss. Xianxin ’bisphosphate binds to hydroxyapatite in the bone and inhibits the bone depletion activity of osteoclasts by its intracellular action. See, for example, H. Fleiseh, Bisphosphonates In Bone Disease, From The Laboratory To The Patient, 4th edition, Academic Press (2000). It has also been reported that bisphosphonates bind to the bone and then release into the depleted cavity during depletion. They are then taken up by osteoclasts and subsequently inhibit farnesyl diphosphate synthase. This intracellular effect, in turn, prevents isopentenylation of the signal transduction protein-GTPase attached to the cell membrane cysts (farnesylation and geranylgeranylation). Geranije The lanylated small GTPase family includes those that guide active bone wasting cells. Please show Zhao, ..., Α · Α · Reszka and G.A. Rodan f, Bisphosphonate Mechanism λ ·

Action ’丨 Curr Rheumatol Rep. 5 (1): 65-74 (February 2003). It is understood that 'bisphosphate has the ability to prevent combined bone loss in some cases > & Know that bisphosphonate has the purpose of preventing bone loss and treatment 彳 not limited to the following diseases. Osteopathy, multiple osteoma, periodontal disease, tooth loss, hyperparathyroidism, rheumatoid arthritis, pagetisdisease, osteonecrosis, osteoarthritis, artificial joint periphery ( peca.kahua) bone loss or osteolysis and cancer guests' + ^ feeding blood disease. All of these conditions are characteristic because of bone loss, that is, bone loss caused by disruption to the formation of * balance with bone f. Alentoric acid is currently the most potent double-money salt available, and it does not destroy bone mineralization at the maximum dose that inhibits bone loss. At present, it has also been found that the increase in bone mineral density found with the administration of quinic acid is positively related to the reduction of spine and non-vertebral (including hip) fractures and the reduction of spinal deformity and height maintenance. This shows that alendurate reduces bone turnover and has a positive effect on producing strengthened bones when administered substantially for a period of time. Alendurate is currently approved in more than 90 countries for the treatment of osteoporosis in postmenopausal women. Alendurate is also approved for the treatment of male osteoporosis, glucocorticoid-induced osteoporosis and skeletal terrible osteitis. Evidence shows that, such as ibandronate, minodr, pamid_ate, rise_te, and tirutoate ( Uludronate) and other bisphosphonates 92852.doc 200538136 salt 'Zoredronate' have many of the same properties as alendurate, including a potent inhibitor of osteoclast bone loss. Despite its therapeutic benefits, bisphosphonates have poor gastrointestinal absorption (approximately 1% grade). See, for example, BJ Gertz et al., Clinical Pharmacology of Alendronate Sodium, Osteoporosis Int., Suppl. 3: 13-16 (1993) and BJ Gertz et al., Studies of the oral bioavailability of alendronate, Clinical Pharmacology & Therapeutics, Volume 58, No. 3, pp. 288-298 (September 1995). I understand that food and many other substances that may be ingested at the same time (including beverages such as mineral water, and even some excipients used to prescribe doses) can adversely affect the absorption of bisphosphonates. Intravenous administration is used to ensure that the complete dose reaches the bloodstream. However, intravenous administration is expensive and inconvenient, especially if the intravenous infiltration that must be administered to the patient is a repetitive occasion and lasts for several hours. Unlike oral administration, intravenous administration of bisphosphonates can be associated with acute kidney injury if administered too quickly. If oral administration is required instead of bisphosphonates intravenously, the agent Xia may be used to compensate for the low bioavailability of the gastrointestinal tract. To offset this low bioavailability, it is generally recommended that patients use bisphosphonates on an empty stomach and fast for at least 30 minutes thereafter. However, many patients find it inconvenient to fast on a daily basis. Bisphosphonate treatment is associated with hypocalcemia. In the case of bisphosphonate treatment: early inhibition of bone loss can induce a small number of blood calcium reductions that occur after the start of treatment: days or days later. The reduction in blood calcium after initiation of treatment can persist for many weeks to months, and may be significant for patients with a deficiency in the vital clock 92852.doc 200538136. Hypoxemia response is sometimes severe enough to be a symptom of bisquamate therapy and requires clinical intervention, especially in patients with hypoparathyroidism (see, for example, Vasikaran, SD, editor., BisphOsphonates: An 〇 guess w oil

Special Reference to Alendronate, Ann. Clin. Biochem. (2001) 38: 608_623). Because there is no substantial physical storage in the body other than bones, the dance required for the formation of new tadpoles after the initiation of tartrate treatment must be absorbed by the diet. If not by food, it is a filling. Vitamin D is necessary for normal calcium absorption. Therefore, for patients using bisphosphonates, adequate vitamin D intake must be obtained. When a net calcium influx into the bone occurs as a result of bisphosphonate treatment when receiving effective osteoporosis treatment, the demand for calcium is ingested for bisphosphonate disease. Adequate vitamin D content is even more important. As a result, patients must have sufficient vitamin D and calcium vitamin D compounds, including those rarely found naturally in foods—groups of fat-soluble open-loops (S are called steroids, and they are found on the skin of vertebrates by the sun UV The light is synthesized by rotation. Although vitamin D may exist in several forms, the most physiologically relevant types are vitamin D3 (cholestyrol) and vitamin ergosterol. The latter are yeast and plant sterols, that is, ergosterol. Exposure to the Koda Shota I as 'as m' originated from 7_dehydrocholesterol, and has a similar metabolic pathway as Picheng, Diaosheng D3 and vitamin ^) 2, and the biological rate in the human body is also Similarly, its main function is to maintain the concentration of tritium in the serum and the normal concentration of the hormone m. 03 as a triol (also known as m, 25_ dicholesterol or 1,25-dimensyl vitamin d3) hormone. Essential precursors whose main role is to enhance the ability of the small intestine to absorb calcium 'and to retain phosphorus in the diet. # 角 晶 醇 之 92852.doc 200538136 The hormonal metabolite is 1,25-dihydroxyergocalcitol (1,25-dihydroxy vitamin 02) ° When the intake of calcium in the diet is insufficient to meet the needs of the body, Parathyroid hormone (PTH), plus calcitriol, a hormone metabolite of vitamin D3, will mobilize monocystic stem cells in the bone marrow to become mature osteoclasts. These osteoclasts are stimulated by various cytokines and other factors to increase the approximate storage of mobilized bones. Cholecalciferol and ergocalciferol in their natural form are non-biologically active precursors of the hydroxylated bioactive metabolites of vitamin D. Because vitamin D is fat-soluble, it is likely to be stored in the adipose tissue of the body, or metabolized into the main storage metabolite of 25-hydroxyvitamin D and stored in other organs. 25-hydroxyvitamin D is transported in the plasma and is metabolized by the body when needed. Specifically, as shown in the implementation of the bile reduced alcohol in Figure 1, 7_dehydrocholesterol in the skin is converted to pro-vitamin 03 (isomers of vitamin VII, Tuxia Wei) when exposed to light. It is converted into a vitamin (cholesterol) after d ^^ Then, cholecalciferol will be metabolized in the liver to form 25 meridian alcohol, also known as meridian vitamin 03; and 'it will progress in the kidney- Metabolized into hormonal form is also known as Ma San Zuo Yi Di Jing _ Huai alcohol. Although, not shown in Figure 1, other metabolites of vitamin d (d2 or D3), including vitamin A, vitamin D, 24, 25, vitamin D, and vitamin D, 1 a, 24, 25i vitamin D . Only matriol has complete production of cholestanol and the metabolites identified above only show very low or vitamin d (d2 & d, ± s). The biological function is to enhance the absorption of calcium in the diet by the small intestine. Efficiency, while chanting the constant of M #,, ,, and temple calcium. It helps to ensure that it is absorbed, 'sufficient in the bow and spoon to maintain the blood word in the positive, the book's circumference' and fully maintain the bone minerals 92852.doc 200538136 D intake promotes absorption in the small intestine, and plays an important role in regulating metabolism and bone mineralization in the eyes. (Month) A w 'Insufficient and deficient vitamin D is the cause of metabolic bone disease in adults. Vitamin D deficiency is characterized by impaired absorption of calcium and phosphorus but normal bone mineralization is not impaired; moreover, the 25. meridian vitamin D content of serum is typically between about 9 to about 30 nanograms / Ml. Vitamin D deficiency is characterized by severely impaired absorption, secondary hyperparathyroidism, hypoxemia, low or impaired blood feeding, and impaired mineralization. Serum vitamin D content is usually about < 9 nanograms / ml. Insufficient vitamin D and deficiency cause increased parathyroid hormone (PTH), which in turn results in increased osteoclast activity, loss of urine phosphorus, and movement of the bones. This in turn aggravates osteoporosis ',', the independence and amount of skeletal strength caused by the mineralization of bones by Tian Wenshou: the elderly when the decrease. We believe that persistent vitamin D deficiency is the main cause of gradual loss of bones and collaterals. Depending on the level of vitamin D and calcium deficiency, the histological picture may be chondrosis, osteoporosis, or a combination of the two. The prevalence of deficiencies and deficiencies of phytobiotics D, which makes patients susceptible to diseases such as osteoporosis; or Yue Wuxu 'and those who receive bisinate therapy for these conditions and absorb extra vitamin E ^ before receiving bisphosphate Salt-treated diseases: I do n’t want to treat patients with insufficient calcium intake or calcium absorption, but fill them with vitamin 0 to promote bone formation and mineralization, and reduce human vitamin D deficiency. It may or may happen. We usually use vitamin B in certain types of bone loss compounds in clinical trials to increase vitamin D intake and recommend: product labeling and product packaging advertising. However, in the United States, for example, about 30% of osteoporosis patients have some level of vitamin D deficiency, and their prevalence increases with the age of 92852.doc 200538136. For our purposes, we recommend that patients receiving oral bisphosphonates who require vitamin D receive two different products at two different times. Vitamin D prescriptions are most often taken daily, and bisphosphonates may be administered daily, weekly, monthly, or longer intervals: the result is' many patients who have been treated for osteoporosis or bone f deficiency, although they have been advised, they cannot Take Vitamin D. Typically, vitamin O cannot be taken at the same time as bisphosphonate because of the fact that bisphosphonate is poorly absorbed; moreover, most oral bisphosphonates are treated orally with bisphosphate and other substances (including ' However, it is not limited to the interval between vitamin D). As a result, there are not many patients who need to administer the vitamin D compound separately at a certain interval in accordance with the dose therapy before or after the administration of bisphosphate. (Some bis-salts need to be taken before ingestion, so any vitamin D administration must be done some time after bisphosphate administration). Although, patients may take vitamin D before or after bisphosphonate doses; in fact, there are evidence that many patients do not. One dating marketing study has shown that although 75-85% of physicians who prescribe alendurate also recommend vitamin supplements, only 57% of osteoporosis patients do adhere. Although it is also possible to administer vitamin D in a multivitamin form; however, for example, when many oral vitamin D formulations are sold in the United States, there is no dosage unit that is less frequent than the daily dose required. In addition, if the patient is administered vitamin D at the same time as the bilinate dose, since many vitamin D compounds prescribed to osteoporosis patients will reduce the absorption of osmium salts, it is likely that the type of vitamin D administered will interfere with and further decrease Absorption of bisphosphate. Current patent literature includes revealing vitamin D3 or vitality buckles, or its 92852.doc 200538136

On behalf of " projectiles or the like, combined patents and published patents for acid reclamation are mostly "poor" such as US patent numbers 4,230,700, 4,330,537 and 4,812,304, European patent numbers EP 0 381 296 and EP 0 162 510 Patent No. WO 90/01321, WO 92/213 55, WO 〇1 / 28564, WO01 / 97788 and WO 03/086415, European Patent Publication No. EP 1 051 976, Japanese Patent Publication Nos. 7_33〇613 and u_6 〇489, U.S. Patent Application Nos. Us 2003/0 ^ 9378 A1 and US 2003/0225039 A1. However, these patents and publications do not disclose or implement less frequently than once a day, but more frequently than once every six months or longer. Compositions, products, or formulations (and, most particularly, lozenges) that can be used for continuous oral administration, such as once weekly at intervals, and most particularly lozenges. These patents and publications also do not disclose or Implemented by administering such bisphosphate / vitamin D compositions less frequently than once a day, but more frequently than once every six months to treat, suppress, reduce or prevent osteoporosis or abnormalities Other conditions of bone wastage. The result is' we need to include a combination of diabolate compounds and vitamin D, including the ability to promote calcium absorption by helping to ensure adequate intake of vitamin D and enhance the overall efficiency of bisphosphate therapy There is also a need to promote normal bone formation and mineralization; at the same time 'reducing or minimizing the occurrence or likelihood of complications such as hypodemia or chondrosis combined with vitamin D deficiency, vitamin D and bisphosphate production ^ It is also necessary to provide a nutritional amount of 3 vitamins and phosphate products that promotes normal bone formation and mineralized vitamin D in patients undergoing treatment with Shuangwei salt. It is also necessary to follow the continuous dose schedule, but at a lower frequency than that 'And more frequently than once every six months or longer. 92852.doc 200538136 Double «salt and vitamin D composition for oral treatment. Also needed, suitable for paper-to-weekly doses can increase bisphosphonate treatment Vitamins at the time: Easy to absorb, and improve the patient's compliance with the recommended vitamin d nutrition including single products of biotin d and bisphosphate. Moreover, it is also a class Method for biotin D / bisphosphate composition. ^ 〃, 鼽 Use this [Abstract] The present invention provides a pharmaceutical composition comprising a bisphosphate compound and a vitamin d compound. Specific embodiments of the present invention include, A pharmaceutically acceptable salt, derivative, or hydrate of a discic acid strong compound 'or a gallate, or a mixture thereof, and a pharmaceutical composition with a vitamin 0 compound such as a pharmaceutical grade vitamin D compound. In a specific embodiment, the vitamin Compound D includes bileuronic acid. In specific embodiments, the salt-filling salts include, for example, arpiturate, alendurate, pharmaceutically acceptable salts (such as sodium, potassium, calcium, magnesium, or ammonium salts, Or any of those salt hydrates) 'such as sporbiturate monosodium salt, alendurate monosodium salt monohydrate, or alendurate monosodium salt trihydrate. In a specific embodiment of the present invention, the pharmaceutical composition comprises cholecholol and alendurate monosodium salt trihydrate. (Refer to Figure 2) The composition of the present invention can be compressed, coated or uncoated tablets, capsules, elixirs, emulsions, or other acceptable dosage forms. In a specific embodiment of the composition of the present invention, the bisphosphate (or a pharmaceutically effective salt, derivative, or hydrate thereof, or a mixture thereof) is present, for example, in a pharmaceutically effective amount from about ⑽ mg to about 1120 mg. In the same or other specific embodiments of the composition of the present invention, the vitamin D compound is present, for example, from about 100 IU to about 60,000 ⑴ of vitamin D compound (40 ⑴ of vitamin D 92852.doc -14- 200538136 quality about 1 Micrograms) of a pharmaceutically effective amount. In other specific embodiments, the present invention relates to a pharmaceutically acceptable salt comprising a vitamin D compound comprising from about 100 IU to about 36,000 IU and from about 5 mg to about 560 mg of a bisphosphate based on a bisphosphonate active basis , Derivatives or hydrates, or a pharmaceutical composition of a mixture thereof. In other specific embodiments, the invention relates to a vitamin D compound comprising from about 1000 IU to about 28,000 IU, and from about 5 mg to about 280

A milligram of a bisphosphate based on bisphosphate activity, a pharmaceutically acceptable salt, derivative or hydrate thereof, or a pharmaceutical composition of a mixture thereof. In other specific embodiments, the present invention relates to a vitamin D compound comprising from about 1,000 to about 8,400 chuan, and from about 5 mg to about 280 mg of a bisphosphate based on a bisphosphate activity, Pharmaceutically acceptable salts, derivatives or hydrates, or pharmaceutical compositions of mixtures thereof. In other specific embodiments, the present invention relates to a pharmaceutically acceptable salt comprising a vitamin D compound from about 100 IU to about 5,600 IU, and from about 5 mg to about 280 mg of a bisphosphate based on a bisphosphate activity. , Derivatives or hydrates, or mixtures thereof. In other specific embodiments, the present invention relates to a vitamin D compound comprising from about 1,000 IU to about 4,200 IU, and from about 5 mg to about 280 mg of a bisphosphate based on a bisphosphate activity. A pharmaceutical composition comprising a salt, derivative or hydrate 'or a mixture thereof. A specific embodiment of the present invention includes about 2,800 IU of vitamin D, and about 70 mg of bisphosphate based on bisphosphate activity, or a pharmaceutically acceptable salt, derivative, or hydrate of bisphosphate, or Its mixed pharmaceutical composition. A specific embodiment of the present invention includes about 2,800 IU of cholecalciferol, and about 70 mg of alenduric acid salt based on suberic acid activity, a medically acceptable of alenturonic acid 92552.doc -15- 200538136 A pharmaceutical composition of a salt, derivative or hydrate, or a mixture thereof. In a further specific embodiment, the cholecalciferol is a pharmaceutical grade. One example of a composition of the present invention is a lozenge comprising sodium alendurate, cholecalciferol, or comprising about equal amounts of cholecalciferol and additional excipients, such as a suitable filler, diluent, adhesive Cholecalciferol particles of binders, lubricants, slippers, disintegrating agents and the like. Further examples of the composition of the present invention are lozenges including sodium alendurate, cholecalciferol, or containing approximately appropriate equivalent amounts of cholecalciferol, lactose, anhydrous lactose, microcrystalline cellulose, colloidal silica, Cholecalciferol granules of sodium carboxymethylcellulose and magnesium stearate. The composition of the present invention can be formulated to cater for different purity and stability. For example, at about < 30 C and about < 30. /. When stored at relative humidity for 24 months, individual isomers (relative to total cholecalciferol) of less than 1% by weight of cholecalciferol are included. The composition of the present invention may also be formulated at about < 30. 〇 and about < 30% relative humidity when stored for 24 months, including less than about 5% total cholecalciferol degradation products, for storage purposes, can be used such as Ming foil package vacuum forming packaging or adding desiccant HDPE Proper packaging of the bottle reduces the effects of environmental humidity on the purity and stability of the composition. In addition, the invention encompasses methods of making the compositions and dosage forms disclosed in this patent specification, and products made according to these methods. A specific example of this method includes preparing a powder mixture including a bisphosphonate such as alendurate, compressing the powder mixture into a mixture, grinding and mixing the mixture with a vitamin D compound to form a final granule mixture And, for example, compressing the final particle mixture to form a spin agent. In a further embodiment of 92852.doc -16-200538136, the final particle mixture can be lubricated before compression. In a specific embodiment ' the powder mixture includes alendurate, colloidal dioxide, anhydrous lactose, microcrystalline cellulose, crosslinked chitomethyl cellulose, and magnesium stearate. In another specific embodiment, the roller of the powder mixture is shrunk to form a compressed strip, which can be ground, mixed with the cholecalciferol particles, lubricated and compressed into a solid dosage form. Advantages of the method of the present invention include increasing the stability of the vitamin D compound of the bilinate / vitamin D composition and increasing the consistency of the particle size of the individual compounds used in the composition of the present invention. Alternatively, the alendurate powder mixture may be pre-mixed with one or more excipients first, then with other excipients, and then compressed by a roller. Other methods of making bisphosphate particles such as, for example, alendurate particles include, but are not limited to, hammering and wet particle methods. If the bisphosphate granules are made by malleting, the bisphosphate (such as quintoate) powder mixture can be compressed into a non-long compact and then ground into granules; it can then be combined with a vitamin D compound Mix to form a particulate mixture that can be subsequently compressed into a solid dosage form, such as a lozenge. Alternatively, the bisphosphate can be wet granulated with all excipients and a granulated liquid (e.g., water), then dried, ground, mixed with a vitamin D compound, and processed into a final dosage form (e.g., a lozenge). In addition to the above methods, a direct mixing method of bisphosphonates, all excipients and vitamin D compounds can also be used, and then compressed into tablets or coated into capsules, or other solid dosage forms. For further description of possible methods for making bisphosphate particles, please refer to US Patent No. 5,358,941, US Patent No. 92852.doc-17200538136 No. 5,882,656 and PCT Publication No. WO 95/29679. It is possible to make a bisphosphate / vitamin D composition as described above and then perform a drying step to reduce the water content of the hydrate. It is also possible to pack humans and desiccants to reduce water content. The invention also encompasses 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, osteoporosis in men, osteoporosis induced by other diseases, unexplained bone & Glucocorticoid-induced Osteoporosis. The present invention also encompasses methods for preventing, reducing, inhibiting or treating osteoporosis, a disease associated with osteoporosis, and other diseases associated with abnormal bone loss. Such other diseases and conditions may include, as further examples, metastatic bone disease, cancerous hyperemia, peripheral hip dissolution, inflammatory joint fires, and others identified as humans and other breast milks. Diseases and conditions. This formula & month is a disease-modifying effect on the induction of joint diseases in mammals. What is it? A therapeutically effective amount of vitamin D / bisphosphate repair is administered to the mammal. The invention also relates to a method for inducing subchondral sclerosis on mammals, preventing fruit formation, preventing the formation or progress of bone spurs, and preventing joint destruction. The method includes administering a therapeutically effective amount of a vitamin D / biscalate combination to mammals. The invention also encompasses methods of reducing the risk of mammalian fractures, including administering an early-dose vitamin D / bisphosphate composition. Such parties, private group human embodiments cover the administration of mammalian hair products including humans.

One thing. Such compositions can be administered at intervals of once a week, once every two weeks, once a month, twice a month, or once every two months. In this method, 92852.doc -18-200538136 vitamin D is provided by the composition of the present invention during the treatment of bisphosphonates while minimizing the occurrence and possibility of complications associated with vitamin D deficiency. According to this, the composition and method of the present invention are useful for mammals identified as having or susceptible to vitamin D deficiency or deficiency, or wanting to have a sufficient amount of vitamin 0. In a specific embodiment, in order to treat osteoporosis and abnormal bone wasting diseases or conditions, and to minimize the risk of secondary diseases due to insufficient vitamin 0, a dose is given once a week, which must be maintained. Until the desired effect is achieved. One of the specific embodiments of the method of the present invention includes administering once a week to Wei Wei suffering from osteoporosis, & containing about 2800 alcohols and about 7G mg of alenduronic acid Salt, or a pharmaceutically acceptable salt, derivative, or hydrate of alendurate, or a lozenge of a mixture thereof. In a specific embodiment, the efficacy of the vitamin D compound administered to the composition of the present invention once a week It is substantially similar to the efficacy of a recommended daily dose of vitamins, such as 4, 0, 600 IU, or 800 IU per day. In addition, this month ’s additional / correspondence determination of the bile of bile acid and bilinate in medicine An alcohol method (such as safety). One example of this method includes' extracting bile alcohol from this composition into the first solution, and forming the first 'heal' from Separation of Cholecalciferol-containing Samples by the Second Solution Reverse high performance liquid chromatography detection of cholestanol in samples, specific embodiments of the method-like method provide increased primary sensitivity, and can be advantageously used in the composition of the present invention to distinguish cholestanol. With pro-alcohols or isomers between _alcohols and pre-alcohols, or detection of bilecohols or pre-coated alcoholic vinegar channels. The invention further encompasses the determination of the administration of the invention Bisphosphonate / cholic calcification 92852.doc 19 200538136 A method for bileducing alcohol in plasma after an alcohol composition. A specific example of such a method includes administering to mammals including alumen salt and cholestrol A composition for obtaining a plasma sample from a mammal, and extracting a cholecalciferol from the plasma sample to form a first solution, and reacting the cholecalciferol of the first solution with a dienophile to form one or more diears of cholecalciferol Diels_ald ^ addition product, the high-performance liquid chromatography (HPLC) separation method was used to separate the dichol-adduct addition product of cholecalciferol, and the mass spectrometer was used to detect the amount of cholecalciferol in the sample. Specific method Sensitivity, and may be beneficial for use in a composition of the invention, for example, to determine the pharmacokinetic effect of administering the composition of the invention. The invention also provides a method for determining the pharmacokinetic effect of elapsed time after the administration of the composition of the invention, including, for example, As reflected in total urine excretion, the serum concentration of lozenges including approximately 70 mg of alendurate and approximately 2,800 m of cholecalciferol versus the area under the time curve (AUC), the maximum at steady state The time (tmax) of the concentration u, Cmax and the median plasma concentration (t1 / 2) are obvious. The present invention may include, exist, or actually include the necessity described herein or the application for a patent scope The ingredients, components, steps, and methods of the present invention as well as the circumstances. Through the following detailed description and practice of the present invention, the progress, specific advantages, specific embodiments, characteristics and different advantages of the present invention will become more apparent. [Embodiment] As used herein, the term "abnormal bone loss" means that the degree of privacy of bone loss exceeds the degree of bone formation, whether locally or to the whole bone, 92852.doc -20- 200538136; or Instead 'bone formation' that would incorporate abnormal structures. The term 'arthritis condition f' or 'arthritis condition, etc.', some of these lesions are inflammatory, and are limited to joints or any inflammation of the joint = condition, the most famous is rheumatoid arthritis (Academic Press Dictionary of Science Technology; Aeademic press, January 15, 1992) ° Arthritis may be caused by inflammation, trauma, or infection. The composition of the present invention may also be used alone or in combination for treatment or prevention Arthritis conditions, or conditions / diseases involving arthritis such as amyloidosis, ankylosing spondylitis, bacterial arthritis, basal calcium phosphate deposits, Behcet's disease, bursitis and tendonitis, CPPD. Diseases, mineralized tendinitis, carpal tunnel syndrome, EhlerS-Dan10s (Congenital Connective Tissue Abnormality Type 4), intestinal arthritis, Fehy, S syndrome, fibromyalgia 'gout, mycotic arthritis, hemorrhage Disease, hemophilic arthropathy, hypertrophic osteoarthropathy, infectious arthritis, inflammatory bowel disease, juvenile arthritis, juvenile rheumatoid arthritis, lupus erythematosus, Disease, Marfan syndrome, mixed connective tissue disease, polycentric reticular histiocytosis, muscle disease, myositis, osteoarthritis, osteonecrosis, osteonecrosis hock disease, polyarterial fire, rheumatism Polymyalgia, psoriatic arthritis, Raynaud's phenomenon, reflex parenchymal insufficiency syndrome, Reiter syndrome, relapsing polychondral fire rheumatoid arthritis, rheumatic fever, sarcomatoid disease, septic arthritis, Scleroderma, Sjogren's syndrome, skeletal dysplasia, lupus erythematosus, and mother arthritis. Unlike rheumatoid arthritis, osteoarthritis is nodule, 'tissue disease, and its pathology is the degradation of joint cartilage due to mechanical injury Reconstruction of bones under the bone and restrictive inflammation of synovial fluid. The net result of these activities 92852.doc 200538136 is joint deformation with secondary damage to articular cartilage wear, cartilage mineralization / spur formation around the joint, secondary joint bones Sclerosis and cyst formation. See Oettmeier, R. and K. Abendroth, 1989, `` Osteoarthritis and bone: osteologic types of osteoarth ritis of the hip, '' Skeletal Radiol. 18: 165-74; Cutolo M, Seriolo B, Villaggio B5 Pizzorni C, Craviotto C? Sulli A. Ann. NY Acad. Sci. 2002 Jun; 996: 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): 23 1-4; Jansson L5 Holmdahl R .; Arthritis Rheum 2001 Sep; 44 (9): 2168-75; and Purdie DW. Br Med Bull 2000; 56 (3): 809-23; see also Merck Manual, 17th edition, pages 449-451 . A specific embodiment of the invention encompasses the treatment, alleviation, suppression or prevention of joint conditions, including the administration of a therapeutically effective amount of a composition of the invention. Another specific embodiment is the treatment, reduction, inhibition or prevention of osteoarthritis, including the administration of a therapeutically effective amount of a composition of the invention. The term "bisphosphate" as used herein corresponds to the following chemical formula:

HO I Ri I OH I 0: 1 = P -II —C ** II _ p — I -0 1 HO I R2 I OH where R! Is independently selected by the following groups: Η, OH and Cl, R2 Independently selected from CH3, CH2, CH2CH2NH2, (CH2) 3NH2, CH2-3-pyridyl, ch2-s-phenyl-C2, ch2ch2n (ch3) (pentyl), ch2-imidazole, CH2-2- 92852 .doc -22- 200538136. Each of the bases + fixed base, N- (cycloheptyl), CH2CH (CH3) 2, (ch2) and CW, and their combinations. In a specific embodiment of the present invention, R1 is 0H, and R2 is a 3-aminopropyl moiety, so the compound produced is 4-aminopyridylbutylene-U-bis-structural acid salt, that is, throtonic acid salt.

The compounds and methods of the present invention also encompass pharmaceutically acceptable salts, derivatives and hydrates of bisphosphates. Non-limiting examples of salts also include those that can only be selected from the following: metal test, earth test, salt and mono, double, triple, or tetrahydrocarbyl substituted ammonium salts, including sodium m and ammonium salts. Non-limiting examples of derivatives include those that can only be selected from vinegar and amidine. The scope of the present invention also encompasses different hydrates and other solvates of the diqin salt, and its medically acceptable salts. The scope of the present invention also encompasses hydrates of alendurate, including but not limited to hydrates having a water content of about M2%, and crystalline forms thereof. Non-limiting examples of hydrates of alendurate and other bisphosphates include monohydrate, dihydrate, trihydrate, hemihydrate, 1/4 hydrate, 1/3 hydrate, 2/3 Hydrate, 3/4 hydrate, 5/4 hydrate, 4/3 hydrate and 3/2 hydrate. Non-limiting examples of difill salts that can be used in this application include the following: Alenduric acid, 4-amino-1-hydroxybutylene-fluorene bisphosphoric acid. Alenduric acid salt (also known as Alenturonic acid sodium or Alenturic acid monosodium salt trihydrate 'or trade name FosamAX®), 4-amino-1-mer Butyl-1,1-bisphosphoric acid monosodium salt trihydrate. (Alentoric acid and alendurate are described, for example, in the following: U.S. Patent No. 4,922,007, published May 1, 1990 to Kieczykowski et al., And U.S. Patent No. 5'019,651 'published May 28, 1991 I Kieczykowski). 92852.doc -23- 200538136 cycloheptylaminomethylene-1,1-bis-carboxylic acid, TM 175, Yamanouchi (incadronate or cimadronate) such as the United States The patent number 4,970,335 is explained by Isomura et al. On November 13, 1990 ° 1,1-digasmethylene-1,1-dilindroic acid (cloturic acid (〇1〇 (11'〇11丨 〇 & (: 丨 (1)) and disodium salt (Cloturate, Procter & Gamble), for example, in Belgian Patent Nos. 672, 205 (1966) and J. Org. Chem 32, 4111 (1967) Explained: 1 _Tyryl-3- (1 _σBilobityl) -Propylene-1,1_Binyl Acid (EB -1 053), 1-Ethyl Ethyl-1,1- Diid acid (etidronic acid), 1-Cyclo-3- (N-methyl-N-pentylamino) -propylene-bisphosphoric acid Also known as BM-210955, Boehringer-Mannheim (Iban (ironate)), for example, described in US Patent No. 4,927,814, published on May 22, 1990. [1-hydroxy-2-imidazole ° pyridine- (l, 2-a ) -3-ylethylene] -bisdonic acid (minodronate). 6-amino-1-hydroxyhexylene-1,1- Phosphoric acid (neridronate) 〇3- (dimethylamino) -1-hydroxypropylidene-1,1-bisphosphate (olpadronate) 〇3-amine -1-Rorylidene propylene-1,1-bisquatic acid (Pamidronate) 〇 [2- (2-Rotyl) -Ethylene] -1,1 -Bisphosphoric acid (piridronate), for example, as described in U.S. Patent No. 4,761,406. 1-Ethyl-2- (3- ° Bis: yl) -Ethylene- 1,1-bisphosphoric acid (Ricea a poor text Θ Wen salt) 〇 0985252.doc -24- 200538136 (4-Gaphenyl) thiolane-i, i-bisphosphate (Tirutoate), such as U.S. Patent No. 4,876,248 was published to Breliere et al. On October 24, 1989. 1-Hydroxy_2- (1 (-imidazol-1-yl) -ethylene_1, 丨 · bisphosphoric acid In specific embodiments of the present invention, the bisphosphate can only be selected from the following: alendurate, pharmaceutically acceptable salts, derivatives and hydrates, and mixtures thereof. The pharmaceutically acceptable salts of alendurate can be made only by the following distillers. Sodium, bell, contract, town, and money salts of arsenate, and can be the monosodium salt of alendurate or its hydrate. Including, for example, sodium alendurate monohydrate 'or sodium alendurate trihydrate. In a specific embodiment, the composition of the present invention comprises sodium alendurate ('Amino ^ Hydroxybutylene Decabis Phosphate Monosodium Salt), which is a member of nitrogen-containing bisphosphate drugs. It should be noted that the terms "bisphosphates" and "bisphosphates", as used herein and directed to the therapeutic agents of the present invention, are meant to also encompass all diphosphates, bisphosphates, and diphosphates, and Salts, derivatives, and hydrates of these materials. Unless otherwise specified, the specific nomenclature used for bisphosphates or bisphosphates is not intended to limit the scope of the present invention, because those skilled in the art are generally familiar with the art. A mixed nomenclature is used, so when a specific weight or percentage of a bisphosphonium salt compound is mentioned in the present invention, unless otherwise specified, it is based on the active weight of the acid. According to its salt, derivative or hydrate type The bisphosphonate dose calculated on the basis is included in the dose range of the present invention based on the active weight of bisphosphonate @ 文 盐. In addition, the dose of all alendurate hydrate types is based on alendurate The basis of the activity 92852.doc 200538136 is 4 different. For example, 'alantoic acid salt and its monohydrate trihydrate, hemihydrate and all other hydrate types are based on : Desire water with active acid weight 4-like aaron melon ten nose. As another example, "On the basis of Asian weight, only alendurate can be calculated. The term “salts, organisms and hydrates, and mixtures thereof, of the gram bone loss-inhibiting bisphosphate” means the choice: The amount of the acid salt compound is based on 7G mg of alentoic acid. Calculate. 'When used in the entire patent specification and special money, please _, " Double Disc㉟

The terms associated with " alantoic acid " include related acid types, pharmaceutically acceptable, and: a balanced mixture of Chinese and some. These terms include crystalline, hydrated, and amorphous forms of alenduronic acid and their pharmaceutically acceptable salts. The term "alantoic acid" specifically includes, but is not limited to, anhydroalantoic acid mononaphthalate, an alantonic acid monosodium salt hemihydrate, an alantoic acid monosodium salt monohydrate, an alantoic acid Sodium salt trihydrate, anhydrous dipotassium salt and dipotassium salt pentahydrate. Alentoric acid monosodium salt monohydrate with other alentoric acid

The crystal form of the fee sodium salt is disclosed in U.S. Patent No. 6,281,381. Sub-pseudo-I potassium salt and its hydrate are disclosed in International Patent Publication No. WO 99/20635. It is customary to give the medicament and calculate the dose of bisphosphonate as much as possible based on the active weight of bis% salt salt, and it is also possible to calculate and use the fine bisphosphate dose according to other salt or hydrate types. For example, the bisphosphonate ritacetate dose is calculated based on the weight of the anhydrous sodium salt of ricate. According to the Doctor's Desk Reference (PhySlcian, s Desk Reference, 55th edition, page 2664 (2001)), for example, each rituxurate tablet contains hemi-pentahydrate plus less 92852.doc -26- 200538136 5 mg or 30 mg of monohydrate form of anhydrous rituxurate sodium equivalents. As used herein, the term "" cholesterol" means that it contains choline alcohol and also contains provitamins. D3, vitamin isomers, granules of vitamin D3 or its isomers and / or additional excipients. The term "continuous progress" or "continuous administration progress" as used herein by Bf means that the administration of the drug is repeated until the desired effect is achieved. Continuous or continuous dosing is different from periodic or intermittent dosing. When used herein, the term "DrT Vitamin 〇3⑽ granules" means gelatin-coated, pharmaceutical-grade Dry vhamin A 100 sold by BASF. When used herein, "general bone loss, The term “means bone loss at multiple skeletal locations or throughout the skeletal system.” The technique of local bone loss is considered to be bone loss at one or more specific defined skeletal locations. When used in As used herein, the terms " humans in need of treatment ", " humans in need of prevention ", " humans in need " and " humans in danger " Humans who are determined to be treated for the disease, who need to be prevented, slowed, inhibited or alleviated, or who are at risk of developing the disease. As used herein, the term "IU" means International Units. When it comes to For the efficacy and dosage of vitamin D, the international unit (IU) is commonly used. An international unit (ιυ) is defined as a crystalline international standard or a pure vitamin of 0 025 microgram specific biological activity To explain in another way, micrograms of vitamins are about 40 international units. 92852.doc -27- 200538136 When used in this article, "mammals that need to be treated", "mammals that need to be prevented", "requires The terms "mammals" and "mammals at risk" mean mammals that need to be treated for the disease, that need to be prevented, slowed, inhibited or alleviated by the clinician or researcher, or have People at risk for developing the disease. As used herein, "one dose per week" means administration of a unit dose 1 ' such as a unit dose of bisphosphonate and vitamin 0 once a week, i.e. once a day

For a long time, it is better to be on the same day every week. In the regimen of one dose per week, the unit dose is usually administered approximately every 7 days. One weekly medication regimen ^ A non-limiting example is a bismuth salt and a vitamin D compound comprising a daily dosage site dose. Weekly application—the second recommended unit dose is conventionally recommended for consecutive days. However, the weekly application—second treatment can include two consecutive days of unit administration during two different weeks. Dosage Therapy As used herein, "bone spur" means a newly formed bone structure located at the edge of the joint 'and its occurrence is strongly related to the recent stage of osteoarthritis. The current hypothesis is that the bone spurs originated from activated periosteum, causing new crescents to overgrow, which eventually turned into bones through the cartilage bone formation process. As used herein, "pharmaceutically acceptable" refers to salts, esters, hydrates, and derivatives of bisphosphates, such as aronite, which means the salts, organisms, or hydrates of the bisphosphate, It has the same general medicinal properties as the free acid form derived from itself and is acceptable from the standpoint of toxicity. As used herein, the term "pharmaceutically effective amount," means when administered according to therapy, such as bisulfate The amount of phosphate compound and vitamin D compound will induce the required therapeutic effect or response of 92852.doc -28- 200538136. A pharmaceutically effective amount of a bisphosphonate is, for example, an amount sufficient to induce prevention, reduction, inhibition or treatment of abnormal bone loss, for example, when administered according to therapy. As used herein, the term "pharmaceutical grade" means sufficient quality and potency to conform to the general conditions of the applicable United States Pharmacopeia (USP) and European Pharmacopoeia (Ph. 彡). Although it is not regulated at this time Formulated USP monographs on vitamin products; however, Ph · Eur_ monographs have been published

Version. Cholecalciferols, for example, are generally better in grade than vitamin D used in general nutritional additives. As used herein, the term "preventing, inhibiting, alleviating or treating," includes the term "directly or indirectly altering the formation or activity of osteoclasts, and dealing with abnormal bone loss (and the resulting physical conditions such as osteoporosis). ; And covers the prevention, suppression, reduction or treatment of bone loss, especially if it is not the mineral phase and / or organic matrix phase, which directly or indirectly alters the formation or activity of osteoclasts

Sex, inhibits removal of existing bones. These terms also mean to treat other disease states or conditions in this manner to promote remission of the disease or condition. As used herein, the term " until the desired effect is achieved " means that a composition such as a bisphosphonate and cholecalciferol is administered at a selected dose schedule, or to follow a course of treatment to a clinician or study for observation The brother asked for a clinical or medical effect on the disease or condition. For the treatment method of the present invention, the bisphosphonate compound can be continuously administered until the desired change in bone Wu 1 or structure is observed. In this example, bone is reached : The increase, to prevent further reduction of bone quality, or to replace abnormal bone structure with more normal bone structure 'are all our desired objectives. Phosphate compounds to prevent unwanted conditions. In this example, the goal is usually to maintain bone density. Non-limiting examples of the application period typically range from about 2 weeks to the end of the remaining life of the mammal. For humans, The application period ranges from about 2 weeks to the end of the person's remaining life, preferably from about 2 weeks to about a leap year, and more preferably from about From 1 month to about 35 years, Yijia is from about 6 months to about 30 years, and most preferably from 1 year to about 20 years. As used herein, the term "vitamin D" means Vitamin A and vitamin 仏 with the chemical structure shown in Figure 3. As used herein, "metabolites of vitamin D" and "derivatives of vitamin 0" mean the metabolites and derivatives of vitamins and vitamin 03. When used herein, "vitamins D compound, the term means vitamins (ergocalciferol) and vitamins (cholcalciferol), 7_dehydrocholesterol and pre_vitamin h and pre_vitamin D3, and 7-dehydrocholesterol, vitamin 仏, Vitamin A, pro-vitamin A, or any isomers or esters of vitamin A, or mixtures thereof. At relevant physiological temperatures, vitamin A and vitamin 达到 reach equilibrium based on their relative previous vitamin isomers, although the balance may be biased toward vitamin h and vitamin Dr. In the present invention, the term vitamin, compound 0, does not Includes metabolites of vitamin D, such as, for example, 25-hydroxycholcalciferol or calcitriol or its analogs; the term also excludes the active hormone calcitriol or its analogs. In this document, vitamin d3 is used interchangeably with cholecalciferol unless specifically stated otherwise. The present invention provides a pharmaceutically acceptable salt, derivative or hydrate of a bisphosphonate, or a bisphosphate, or a combination thereof with a vitamin] 3 compound 92852.doc -30-200538136. In an exemplary embodiment, the bisphosphate compound is selected from the group consisting of sodium alendurate, sodium alendurate monohydrate, or sodium alendurate trihydrate, and the vitamin D compound is cholestanol. . The exact dose of the bisphosphonate and vitamin D compound will depend on the progress, oral efficacy of the particular bisphosphonate selected, the mammalian or human

Gender and illness, the severity of the condition to be treated, and other relevant g-physical and physiological factors. Μ ′ cannot be accurately identified in advance. The effective amount 1 can be easily determined by the caregiver or clinician. Based on animal models and clinical studies in humans, routine experimentation can determine the appropriate ^. Generally, the pharmaceutically effective amount of bisphosphonate is selected from the continuous administration schedule until the desired effect is achieved. For humans, an effective oral dose of bisphosphonate typically ranges from about 0.0001 mg / kg body weight to about 100 mg / kg body weight 'for a 75 kg patient', preferably from about 0.000 mg / kg body weight. 0.05 to about gram / kg body weight. In a specific embodiment of the present invention, an appropriate amount of a vitamin D compound is selected to provide sufficient vitamin D nutrition during the administration interval without interfering with the ability of the bisphosphate to obtain the effect of inhibiting bone loss. For oral doses of the present invention including alendurate, pharmaceutically acceptable salts, derivatives or hydrates of alendurate, or mixtures thereof, and vitamin D compounds, the amount of vitamin D compounds includes About 100 IU to about 60,000 IU. A non-limiting example of an oral amount of a vitamin D compound in a specific embodiment of the present invention is' including but not limited to 700 iu, 1, 400 IU, 2,800 IU, 4,200 IU, 5,600 IU, 7,000 IU , 8,400 IU, 14,000 IU, 28,000 IU, 36,000 IU, and 60,000 111 doses of vitamin 1) compounds. 92852.doc -31-200538136 An oral combination comprising a vitamin D compound and a pharmaceutically effective amount of alendurate, or a pharmaceutically acceptable salt, derivative, or hydrate of alendurate, or a mixture thereof In terms of properties, the pharmaceutically effective amount of alendurate typically includes from about 0.05 mg to about Π20 mg of a mesostate compound based on the weight of alenduric acid. In a specific embodiment of the present invention, non-limiting examples of an orally pharmaceutically effective amount of alendurate include, but are not limited to, about 2.5 mg, 5 mg, 8.75 mg, 10 mg, 17-5 mg, 35 mg, 4 〇 House grams, 70 mg, 140 mg, 280 mg, 560 mg, and 1120 mg_ Alenturonic acid salt dosages based on the weight of alenduric acid each. The bisphosphonate and vitamin D compositions of the present invention are typically formulated according to the dosage form for oral administration, and are typically mixed with appropriately selected pharmaceutical diluents, excipients or carriers. Examples of oral dosage forms include lozenges (including compressed, coated or uncoated), capsules (each of which includes sustained release or timed release), hard or soft gelatin capsules, tablets, pills, powders, granules, ugly fluid , Citron, clay, foamed tablet composition, film, sterile solution or suspension, syrup and emulsion, and the like. Similarly, it may also be administered intravenously (giant pill or infiltration), intraperitoneal collapse, topical (such as eye drops), intranasal, inhaled, subcutaneously, intramuscularly, or transdermally (example, tablet). 〉 Glutamate or liquid sprays, drops, ampoules, automatic injection devices or all types of pharmaceutical techniques known to those skilled in the art. Effective but non-toxic amounts of the desired composition can be used. The composition is administered orally, parenterally, intranasally, sublingually or rectally, or for inhalation or oral administration. Popular administration. According to the prescription of the composition of the present invention, it can be conveniently carried out by the method known in the art, for example, as described in Remington, pharmaceutical Sciencesl, 17th edition, 5 years. 92852.doc -32- 200538136% for oral administration. For example, for tablets, capsules, the active ingredient can be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as lactose, starch, strict sugar, Glucose, methylcellulose: magnesium stearate, mannitol, sorbitol, cross-linked sodium methylcellulose and the like; liquid types for oral administration, such as liquid contact, syrup, mud, emulsion, suspension Liquid, solution and foam bond compositions can combine the oral pharmaceutical ingredients with any oral, non-toxic, pharmaceutically acceptable inert system such as ethanol, glycerol, water and the like. Moreover, when necessary or necessary, appropriate adhesives, fillers, diluents, turbidizers, auxiliaries, disintegrating agents, buffering agents, coating agents, and coloring agents may be incorporated. Suitable adhesives include, but are not limited to, gluten flour, gelatin, natural sugars such as glucose, anhydrous lactose, / 9-lactose, corn sweeteners, such as natural and synthetic gums such as gum arabic, and steamed gum. Sodium, Jun methyl cellulose, polyethylene glycol, ants and the like. The lubricants used in these dosage forms can include, but are not limited to, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium acetate and the like. Suitable decomposing agents may be one or more modified cellulose powders / cellulosic polymers ' including cross-linked sodium methylcellulose. Diluents that can be used as a shrinking agent include, but are not limited to, lactose, diphosphate, cellulose, cellulose, and the like. In the present invention, a slip agent having a flow characteristic of a powder blend can also be used. Examples of the slip agent include 2㈣ '«dioxy_, talc and the like. The composition used in the present invention f can also be coupled to a soluble polymer as a target: a fun carrier. Such polymers may include, but are not limited to, polyethene, "copolymers," polypropylmethamphetamine, and the like. 92852.doc • 33-200538136 Additional excipients such as those described in U.S. Patent No. 5,358,941, U.S. Patent No. 5,8 82,656 and PCT Publication No. WO 95/29679 can also be used. A specific embodiment of the present invention, for example, comprises about 05% to about 90% by weight of sodium alendurate, and about 1% to about 70% by weight of cholecalciferol particles (equivalent to about 0.0005% to about 20%). % Of cholecalciferol), about 10% to about 80% by weight of anhydrous lactose, about 5% to about 50% by weight of microcrystalline cellulose, and about 0.1% to about 5% by weight of colloidal silica , About 0.5 to about 10% by weight of croscarmellose sodium, and about 0.5 to about 5% by weight of magnesium stearate in 80 mg to 1500 mg lozenges.

The weight range of Dry Vitamin DJOO granules is to ensure a potency of 2800 IU per lozenge 'because the granules contain from 100,000 to about 110,000 IU of vitamin D3 per gram. Anhydrous milk bran is adjusted based on the tablets of Dry Vitamin D "〇〇 added to maintain a final tablet weight of 325 mg. Other non-limiting examples of oral compositions of the present invention include bisphosphate compounds such as herein The illustrated alendurate and vitamin D compounds include those described in the following examples. Dry Vitamin D3100 granules contain approximately 100,000 IU of vitamin D3 per gram of particle weight. Therefore, 28 mg of Dry Vitamin D3100 granules contain approximately 2800 IU of vitamin D3, It is about 70 micrograms of vitamin D3 equivalent. The double salt salt / vitamin D composition of the present invention includes the specific embodiments described herein, but the application interval is once a week, once every two weeks, once a month, Twice a month and once every two months. For a composition of the present invention administered once a week, a pharmaceutically effective amount of alendurate includes an active weight basis based on Aaron 92852.doc -34- 200538136, Alentorol® from about G.05 mg to about 112 mg. Specific embodiments of the present invention provide a pharmaceutically effective amount of a quintoate once a week including, but not limited to, useful Prevention of osteoporosis: dosages, including vitamin D compounds, and alendurate compounds from about milligrams to about 70 milliliters; unit doses that can be used to treat osteoporosis include 'vitamin D compounds and about 70 mg of alendurate compound; unit doses that can be used to treat teratogenic osteitis, including vitamin D compound m8G mg of Argentine 8 man salt compound; and unit doses that can be used to treat transfer bone disease, including Vitamin D compound and about 280 mg of alendurate compound. A pharmaceutically effective amount of a vitamin D compound of the bisphosphate / vitamin D composition of the present invention includes from about 100 IU to about 60,000 for one dose per week. IU vitamin d. According to this, in a specific embodiment of the present invention, the composition includes a vitamin D compound from about 100 m to about 5,600,000, and a pharmaceutically effective amount of alendurate, aron A pharmaceutically acceptable salt, derivative, or hydrate of a tartrate salt, or a mixture thereof. In another specific embodiment, a pharmaceutically acceptable amount of the alendurate salt includes, from about 0.1. 5 mg to about 1120 mg of alendurate based on the active weight of alenduric acid, a pharmaceutically acceptable salt, derivative or hydrate of alendurate, or a mixture thereof. For two weeks or two For a single dose, the pharmaceutically effective amount of the vitamin j) compound of the bisphosphate / vitamin D composition of the present invention includes from about 100 IU to about 60,000 IU of vitamin D. In a specific embodiment of the present invention The composition includes a vitamin D compound from about 100 IU to about 8,400 IU, and a medically effective amount of alendurate, a medically acceptable salt and derivative of alendurate, 92852.doc -35- 200538136 Or a hydrate, or a mixture thereof. In another specific embodiment, the pharmaceutically acceptable amount of the alenduric acid salt comprises, from about 0.05 mg to about 1120 mg, the alenturonic acid salt based on the active weight of the alenduronic acid, the alendonic acid A pharmaceutically acceptable salt, derivative, or hydrate of a tartrate salt, or a mixture thereof. For one dose per month, a pharmaceutically effective amount of a vitamin D compound of the bisphosphate / vitamin D composition of the present invention includes from about 100 μv to about 36,000 IU of vitamin D. In a specific embodiment of the present invention, the composition comprises a vitamin D compound from about 100 IU to about 11,200 IU, and a pharmaceutically effective amount of alendurate, a pharmaceutically acceptable amount of alendurate Salt, derivative or hydrate 'or a mixture thereof. In another specific embodiment, the pharmaceutically acceptable amount of the alenduric acid salt comprises, from about 0.05 mg to about 1120 g of alendurate salt, based on the weight of the active ingredient of alenduronic acid, alendonic acid. A pharmaceutically acceptable salt, derivative, or hydrate of a tartrate salt, or a mixture thereof. The invention also encompasses the prevention, alleviation, suppression and treatment of diseases and conditions associated with abnormal bone loss such as osteoporosis. A person suffering from osteoporosis, that is, a bone mineral density (BMD) with a standard deviation distance of at least about 2 or less than 2 1/2 of a normal premenopausal woman 'is a candidate for applying a sclerosing composition according to the method of the present invention. It was found that those who administered a single dose of vitamin 03 7 times or more compared to those administered on a daily basis could co-administer bisphosphonates such as alendurate, without Negatively affects the bioavailability of bisphosphate. See, for example, Example 7. The method of the present invention does not have the disadvantages that current methods of treatment require complicated, irregular, or complex drug delivery therapies in order to provide adequate vitamin d when followed by bisphosphonate treatment. 92852.doc 200538136 As a result, for a composition of the present invention including, for example, a combination of a bisuplate compound and a vitamin D compound, such as alendurate, for all compositions including alendurate or other dipicric acid Salt without vitamin D compounds will be effective for all indications. The method of the composition of the present invention can be used to reduce or inhibit bone loss, and to treat, reduce, inhibit and prevent abnormalities. 1 = Pelvic or a combination thereof. Therefore, the composition of the present invention can be used in / and other animals to increase bone quality and prevent, inhibit, reduce and treat the following conditions and disease states: bone loss, osteoporosis, including but not limited to postmenopausal osteoporosis Disease, steroid-induced osteoporosis, male osteoporosis, disease-induced osteoporosis, pathologically-induced osteoporosis, and glucocorticoid-induced osteoporosis, osteonecrosis, aberrant osteitis, osteoarthritis, and similar Rheumatoid arthritis, other joint conditions, abnormally increased bone turnover, local bone loss associated with bone loss or osteolysis around pro-articular joints, fractures, metastatic bone disease, Gaucher's disease, avascular necrosis, Multiple osteomalacia, Charcot Joint, parasitic disease, osteogenesis imperfecta, homohematomic aciduria, urinary protein intolerance, occupational disease, mobilization, fiber Metaplastic progressive ossification, fibrotic osteogenesis imperfecta, periodontal disease, tooth loss, hyperdemia of cancer, multiple myeloma, scarcity of bone, including But it is not limited to mobilization-induced bone scarcity and bone scarcity due to bone metastasis, and other bone diseases and conditions that may be associated with abnormal bone loss.

The present invention relates to the preparation of a medicament for treating, reducing, inhibiting or preventing joint diseases by using the composition of the present invention. The present invention also relates to the use of the composition of the present invention and an enzyme inhibitor selected from the androgen receptor modulator, osteoclast proton ATP 92852.doc -37- 200538136, HMG-CoA reductase inhibitor, osteoblast assimilation agent, Oralin, vitamin K2, or pharmaceutically acceptable salts and mixtures thereof are used in the preparation of medicaments for treating joint diseases. In a specific embodiment of the present invention, the joint condition is powder-like degeneration, ankylosing spondylitis, bacterial joint i, basal filling and crystal accumulation disease, Bessie's disease, bursitis and tendinitis, and cppD accumulation. Disease, mineralized tendinitis, carpal tunnel syndrome, Ehlers_DanlQs (congenital connective tissue type 4), intestinal arthritis, secretory syndrome, fibromyalgia, gout, fungal arthritis, heme disease, Hemophilic arthropathy, hypertrophic osteoarthropathy, infectious arthritis, inflammatory bowel disease, juvenile arthritis, juvenile rheumatoid arthritis, lupus erythematosus, Lyme disease, Marfan syndrome, mixed connective Tissue disease, polycentric reticular histiocytosis, myopathy, myositis, osteoarthritis, osteonecrosis, osteonecrosis hock disease, polyarteritis, rheumatic polymyalgia, psoriasis arthritis, Raynaud's phenomenon, Reflex sympathetic insufficiency syndrome, Reiter syndrome, relapsing polychondritis, rheumatoid arthritis, rheumatic fever, sarcomatoid disease, septic arthritis, scleroderma, Sjogren syndrome, bone Skeletal dysplasia, lupus erythematosus, and viral arthritis. A specific embodiment of the present invention is a method for treating, alleviating, inhibiting, and preventing osteoarthritis in a mammal in need thereof, including administering to the mammal a therapeutically effective amount of a composition of the present invention. The well-defined changes associated with osteoarthritis are known in the literature, including articular cartilage surface wear, articular cartilage mineralization / long bone spurs, and subchondral bone sclerosis and cyst formation. See Oettmeier R3 Abendroth, K5 " Osteoarthritis and bone: 92852.doc -38- 200538136 osteologic types of osteoarthritis of the hip ,, Skeletal Radiol 1989; 18: 165-74. Recently, the role of subchondral osteosclerosis in initiating the osteoarthritis process has been pointed out. When the joint responds to repeated compression and destruction of the cartilage and bone, it is less able to reduce or disperse the force through the joint, causing it to withstand greater mechanical pressure on the surface of the articular cartilage. This in turn accelerates cartilage wear and fibrosis. Refer to Radin, EL and Rose RM ,, Rollef subchondral bone in the initiation and progression of cartilage damage, "Clin. Orthop · 1986; 213: 34-40. By the composition of the present invention to inhibit the secondary articular bone Attrition will cause inhibition of subchondral skeletal transition, and therefore have a positive effect on the process of osteoarthritis. Another embodiment of the present invention is a method for treating, reducing, inhibiting or preventing the condition of mammalian rheumatoid arthritis. Including administering a therapeutically effective amount of the composition of the present invention. It is known in the literature that continuous destruction of bones around joints is the main cause of joint function destruction and loss of capacity in patients with rheumatoid arthritis. See Goldring SR ,, Pathogenesis of Bone erosions in rheumatoid arthritisifCurr. Opin. Rheumatol. 2002; 14: 406-10. In addition, generalized bone loss is the main cause of morbidity rates associated with severe rheumatoid arthritis. The frequency of hip and spine fractures is actually Increases with chronic rheumatoid arthritis in patients. See Gould A. Sambrook, P, Devlin J, et al., ' Osteoclastic activation is the principal mechanisn leading to secondary osteoporosis in rheumatoid arthritis, "J. Rheumatol 1998; 25:. 1282-9. Anti-wear agents used in the treatment or prevention of sub-joint bone loss and general bone loss represent a reasonable way for medicine to interfere with the process of rheumatoid arthritis 92852.doc -39- 200538136. Accordingly, the composition of the present invention including a bisphosphate compound and a vitamin 0 compound can be used to treat, reduce, inhibit or prevent bone loss associated with rheumatoid arthritis and other osteoarthritis conditions. More commonly, it is believed that bisphosphonates can be prescribed with vitamin D without adversely affecting the bioavailability of bisphosphate. Furthermore, people believe that low-dose vitamin D and high-dose vitamin D can be prescribed with bisphosphate 'without negatively affecting their bioavailability. As one example, it is believed that m-vitamin D, measured once-weekly, is effective when administered in combination with bisphosphates in the present invention. It has also been known that vitamin D2 can be replaced with vitamin D2 to have similar effects to those found in vitamins. Therefore, 'administration of a vitamin D compound on the same prescription as a bisphosphate compound will reduce vitamin d which should be administered separately and provide vitamin D nutrition' during the treatment of bisphosphonates without adversely affecting the biological activity of bisphosphonates. Utilization and efficiency. Patients will benefit from the combination of vitamin D and bisphosphonates, as it improves extra vitamin D nutrition that promotes normal bone formation and mineralization, and enhances the efficiency of dual-lin / alpha therapy. From the patient's lifestyle and compliance point of view, the method of the present invention is more convenient than administering bisphosphonates with an additional dose of vitamin D per or periodic administration. As a result of the present invention, since the present invention provides a once-a-week dose of vitamin D, patients no longer need to take daily vitamin D separately to benefit from additional vitamin d nutrition. The patient does not need the complex meal therapy of double meal salt treatment and vitamin D administration without the need to be separated. In the end, the patient will resort to the inconvenience of having to take a biquatate mouthpiece on an empty stomach 'and will have to fast for at least a few minutes before and after administration. Because of the 92852.doc -40-200538136, the method of the present invention may have the advantage of promoting better patient compliance; moreover, this may in turn be converted into a better therapeutic effect. It is also believed that bisphosphate / vitamin D compositions are also less irritating to the esophagus and gastrointestinal system. Because alendurate has the potential to penetrate the layered horny epithelium, the base layer of field cells (for example, through its own penetration or through its teeth, localized injury sites caused by abrasive foods or other agents) ) 'It will cause inhibition of keratinocyte growth, such as its effect on keratinocyte growth in vitro. See AA Reszka et al., MoI Pharmacoll, 2001; 59 (2) · 193 202. Inhibition of growth may delay the process of epithelial cell repair, thereby causing local irritation or ulcers. Autoradiography of rats fed with # 125 (0 ^ 2 vitamin h did indeed show evidence of vitamin D receptor expression in esophageal epithelial cells. See Stumpf, WE, et al., Histochemistry, 1987; 87 (1) : 53-8. The amount is lower than that seen in the parathyroid. Therefore, it is believed that co-administration of active vitamin d3 metabolites (such as 1,25 (ΟΗ) 2-cholcalciferol or calcitriol) and alenduronic acid Salt will aggravate the esophagus through its differentiation effect on keratinocytes. It has been observed that both calcitriol and 2 5 -OH-bile # 5 alkanol can affect keratinocytes by inhibiting growth and inducing differentiation. See K. Matsumoto et al., Biochem. Biophys. Acta., 1991; 1 092 (3): 3 1 1-8. Keratinocyte differentiation is associated with cell cycle arrest (growth arrest), so alendurate is an active vitamin D metabolite The combination has a synergistic effect on the inhibition of grassroots growth. This in turn will cause a greater stimulating effect. Because oral vitamin D3 (cholcalciferol) needs to be activated in the liver and then in the kidney, it is believed that it will not be as active metabolites Induces the same local stimulating effect. 2.doc -41-200538136 In addition, the normal physiological amount of active vitamin d3 hormone may help the body try to repair local irritation sites damaged by acute exposure to alendurate. Oral vitamin D3 is combined with alendurate Administration, and because a large proportion of the elderly population lacks vitamin d3, this may allow the body to speed up the healing process. It is also believed that vitamin D / bisphosphate compositions can be used to prevent or treat skewness. In addition, people also believe that vitamin D / Bisphosphate compositions can be used to reduce falls. It is also believed that vitamin D / bisphosphate compositions can increase muscle strength, improve neuromuscular function, reduce body skew and improve physical function in the elderly. This will reduce the risk of falls, and It therefore contributes to the reduced risk of fractures. Epidemiological studies show the prevalence of vitamin D deficiency in the elderly population in the United States, with reference to AN 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 also evidence that vitamin D has an effect on extraskeletal tissues. Refer to Latham et al., 2003; 51: 1219-1226. In addition, vitamin D receptors have been identified in muscle tissues, and muscle weakness, limb pain, and physiology Impaired function is evidence that I can recognize the lack of vitamin D. Some prospective, opportunistic intervention studies have shown that vitamin D is effective in improving musculoskeletal function and reducing the risk of falls. Treatment with vitamin D and calcium has been shown to reduce the number of cases of non-vertebral fractures, as well as reduce postural skew and possible falls. See LK. Dhesi et al., Age and Aging 2002; 31: 267-271. In addition, it has been shown that the number of falls in elderly residents living in the community is 92852.doc -42 · 200538136

Objective: It can be significantly reduced due to the benefits of alfacalcid '1βα, a base vitamin called the treatment and the minimum intake. Refer to L. Dukas et al., JAGS 2004; 52: 230-236. People also believe that vitamin D / Bisphosphonate composition can enhance the absorption of calcium. Use / tongue | ± vitamin D metabolites to examine its positive effect on partial calcium absorption in postmenopausal women. Refer to ML · Holzherr et al., Osteoporosis Int 2000; 11: 43-51; J c and others,] d

Endocrinol Metab, 1980; 51 (5): 1359_64. Bisphosphate has been shown to increase calcium absorption in the small intestine in rat models. With reference to p. Ammann et al., 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, it is believed that the combination of a vitamin 0 compound and a bisphosphate compound may have an additive effect on the absorption of calcium when vitamin D or bisphosphate is administered alone. In addition, it is also believed that a combination of cholecalciferol and alendurate will greatly increase the absorption of calcium compared to the combination of active form of vitamin D and a different diacetate. This increase in calcium absorption is positively related to a reduction in fracture risk. The invention also provides the use of a composition comprising a vitamin D compound and a bisphosphate compound, 'including a pharmaceutically effective amount of at least one bisphosphate, or a bile salt, derivative or hydrate, Or mixtures thereof with one or more active ingredients to produce a medicine that treats, reduces, inhibits or prevents the above-identified conditions and disease states such as human mammals. In a further specific embodiment, the method and composition of the present invention can also include 92852.doc -43- 200538136 including a histamine H2 receptor blocker (that is, an antagonist) and / or a proton pump inhibitor, which are Agents known to increase gastric pH. See, for example, L J Hixson, et al., Current Trends in the Pharmacotherapy for Peptic Ulcer

Disease, Arch · lntern · Med ·, Vol. 152, pp. 726-732 (April 1992). It has been found in the present invention that sequential oral administration of histamine receptor blockers and / or proton pump inhibitors, followed by the addition of bisphosphate and vitamin D compositions, can help to minimize the negative effects on the gastrointestinal tract . In a specific embodiment of the present invention, the histamine H2 carcass is administered from about 30 minutes to about 24 hours, or from about 30 minutes to about 12 hours before the application of the bisphosphate and vitamin D composition. Blockers and / or proton pump inhibitors. The dosage of the histamine receptor blocker and / or proton pump inhibitor will depend on the particular compound selected and the mammal being treated, i.e. size, health and other factors. Non-limiting examples of histamine H2 receptor blockers and / or proton pump inhibitors include those that can only be selected from: cimetidine, famotidine, nizate Nizatidine, ranitidine, omprazole, and iansopraz (a). The present invention further encompasses methods of making the compositions of the present invention, including, for example, including bisphosphate compounds and A pharmaceutical composition of a vitamin D compound. In a specific embodiment, 'It is a method for preparing alendurate-cholesterol formulation' includes preparing a powder mixture including alendurate and compressing the ground mixture to form Alendurate mixture, which is ground and mixed with cholecalciferol particles to form a mixture, and lubricates and compresses the mixture. In another embodiment, it is to prepare alendurate_ Cholecalciferol solid dosage method, including mixing alendurate, colloidal dioxide 92852.doc -44- 200538136 Shi Xi, anhydrous lactose, microcrystalline cellulose, and croscarmellose sodium A premix is formed, the premix is mixed with magnesium stearate to form a first lubricating mixture, and the roller compresses the first lubricating mixture to form a compressed bar. 'The compressed bar is mixed into a lubricating mixture, which lubricates The mixture is mixed with cholecalciferol particles to form a second lubricious mixture, and the second lubricious mixture is compressed into a solid dosage form.

Figure 4 illustrates a flow chart of a specific embodiment of the method of the present invention for making a bisphosphate / vitamin D composition. In this specific embodiment, the composition is manufactured by the following methods, including roller compaction of sodium alendurate to form a bar, grinding the bar produced by the roller compaction step, and then mixing with the vitamin h prescription Granular additives. Utilizing, for example, the active ingredients and excipients identified in Example i, the method and method here produce a product that can satisfy the regulation of the release of both alumenic acid salt and vitamin A and a product that is stable and stable as shown in Figure 4. In this specific example, a sodium premix of colloidal silica, anhydrous lactose and alenduronic acid is prepared in step% 1. As described in step 302, the premix is then mixed to microcrystalline cellulose and crosslinked sodium methylcellulose. In the step, a stearic acid lock is added to form a lubricious mixture. As shown in steps 3G4, the lubricating mixture is passed through a roller compressor and the compressed bars are ground. In the specific embodiment shown in FIG. 4, then in step 305, vitamin granules containing 2 μm (or about 7 (micrograms) dangshen vitamin 仏) are added, and the grinding granules are analyzed according to the particle analysis of the feed and the roller compression / The yield in the grinding step is mixed with the vitamin D3 granules filled by hand. Then, in step 3, the spleen &# / 娜川 6 is used to compress the resulting mixture to form a tablet, and shrink the tablet. Agent to remove dust. Keli J 扪 uses, for example, waterproof and opaque 92852.doc • 45- 200538136 vacuum forming packaging or bottles of suitable packaging materials to form the key bonding agent. Vitamin d3 (cholesterol) and vitamins Sheng Jiao (ergot) is a water-insoluble, hydrophobic compound with dazzling points of about 8 generations and about n5t. These compounds are also highly susceptible to oxidation and are easily damaged by light, and are broken into different degradation products. Vitamin D particles are also It is easy to separate. Therefore, the stability of vitamin 0 is affected by the degree of defense and properties of the vitamin D / bisphosphate composition and the storage conditions (such as exposure to light, high temperature and high relative humidity). As a result, the desire to include vitamin D in the composition of the present invention presents a special challenge in terms of 'the production and storage of vitamin D-containing compositions which are of interest to us. Accordingly, a vitamin D / bisphosphate composition that reduces vitamin D degradation during processing or storage that requires a prescription. There is also a need for a method for making such stable compositions. In addition, there is also a need to develop pain testing or assays for the degradation of vitamin D-containing compositions such as those shown in the present invention. In addition, because the amount of specific vitamin D degradants may be very low (nanogram grade), it is necessary to develop a quantitative limit (LOQ) that is sufficient to detect the amount of vitamin D degradants. Method for degradation of vitamin 0 in a vitamin D-containing composition. Accordingly, the present invention also provides a method for producing a composition including a bisphosphate compound and a vitamin D compound, which can minimize the loss of vitamin D during manufacture. By controlling the humidity at the time of manufacture, the temperature and light at the time of prescription of vitamin D ', or the provision of a suitable finished dosage form, it is possible to reduce the loss of vitamin D' while maintaining its full efficacy. In a specific embodiment of the present invention, the temperature in the granulation is lower than or equal to about 35 ° C. In a further specific embodiment, the temperature is between about 2 ° C. and about 30 ° C. In a specific embodiment, the relative humidity in the 92852.doc -46- 200538136 process is less than or equal to about 60% RH. In another specific embodiment, the relative humidity is between about 20% to about 40%. In addition, the control will target the initial humidity of the vitamin D ingredient of the prescription and also any excipients that may appear. In a specific embodiment, the relative humidity is between about 25% and about 35%. The present invention also includes a manufacturing method including an additional drying step. Therefore, as another specific embodiment, the composition of the present invention may be used as described above. Manufactured under different conditions (temperature and / or relative humidity), and the water content of the resulting composition can also be reduced by drying the composition. In another embodiment, the drying may involve the production of a solid dosage form The composition of the present invention is dried (eg, by thermal energy). In another embodiment, the drying process may involve film coating a solid dosage form (eg, a lozenge) of the composition of the present invention. In another embodiment, For example, the drying may also involve packaging the composition of the present invention with an appropriate amount of a desiccant or other portion to reduce water content. In another specific embodiment, the drying may involve packaging the composition of the present invention to Reduced moisture and / or light storage type storage (such as aluminum vacuum forming bag, waterproof bottle). In the specific embodiment of the present invention, the vitamin D compound used as the starting material may include free flowing, stable Vitamin D granules. In a specific embodiment, the vitamin D granules used as a starting material in the manufacturing method of the present invention are medical gelatin-coated pharmaceutical grade 3 1 100.δ vitamin D granules commercially available from BASF. It is dissolved in medium-chain triglycerol with 1-2 micron droplets. It is embedded in the gelatin and sucrose matrix coated with the powder. Then, the dissolved vitamin D can be stabilized with tertiary butylhydroxybenzene (BHT). The vitamin 92852.doc 200538136 D granules contain sodium aluminum oxalate as a flow aid. Those skilled in the art will understand that the amount of vitamin D added to the composition may need to be maintained Source and / or the effectiveness of added vitamin D. For example, if gelatin-coated pharmaceutical-grade Dry Vitamin D3 100 (BASF) is used, those skilled in the art will understand that the effectiveness of the particles may be different (Such as 100,000 IU / g or 105,000 IU / g or 110,000 117 / g), it may be necessary to adjust the amount of particles added to the composition so that the vitamin d of the composition reaches 2800 IU or 5600 IU. In a specific aspect of the invention Example, the vitamins contained in it are consistent with the acceptance criteria of 1 > 11 · Em cholecalciferol concentrate monograph (powder type). Although, at this time, there is no prescription monograph on vitamin A products for vitamin A products, it is true A monograph on Ph.Eur. Cholecalciferol Concentration was published. Inactive ingredients of the vitamin D compound composition used in specific examples of the composition of the present invention (for example, medium-chain triglycerides, third butyl hydroxytoluene, sucrose, gelatin, modified starch, and sodium aluminum silicate) If it is not rough, it is food-grade material. In the specific embodiments of the composition and method of the present invention, the alendurate used as a starting material is a rough grade of alenduronic acid sodium monohydrate, or it is purchased from Merck & Co., Inc. Rough grade sodium alendurate trihydrate. In addition, it is also possible to use in the compositions of the present invention, such as commercially available vitamin o granules, which are commercially available from Roche, BASF or Solvay. In a further embodiment, the present invention provides a kit that can be easily and efficiently implemented the method according to the present invention. Such kits are particularly suitable for delivering solid oral forms such as lozenges or capsules' and include, in specific embodiments, one example of a unit having a dosage of 92852.doc 200538136 according to its intended use is "vacuum packaging," Known for kits, and widely used in ... M-shaped packaging is a memory aid familiar to packaging technology, such as a number, the letter 22 can be provided, indicating the dose that can be applied ~ = or other labels or layers The date of the progress of the / D treatment in the vehicle insertion medium ①. Alternatively, when providing a high-volume kit, it may include a significantly different type of placebo if it is not similar to the early dose of bisphosphate d, Or Tension Supplement. In those embodiments that include group receptors and / or proton pump inhibitors, these agents can be included as part of a kit. The present invention also provides for the determination of vitamin A in the composition of the present invention Determining methods developed by degradation products. Specifically, the method for determining the degradation of the pharmaceutical composition may include 'extracting a bile alcohol from the first composition into a first solution to form a second Cholecalciferol-containing sample is separated from the second solution, and the content of cholecalciferol in the sample is detected by reverse HPLC separation of the sample. The detection method of the present invention is to detect the approximate concentration of each pharmaceutical composition. Cholecalciferol from 2800 IU to about 5600 IU. In addition, the detection method has a cholesteryl alcohol quantification limit (10q) of less than about 9 μg / μL of cholestyrol. In a specific embodiment, the The method utilizes a first solution that includes water, ethanol, acetonitrile, or a mixture thereof. In a specific embodiment, the first solution includes about 5% water and about 95% methanol. The sample formulations exemplified may be 15% Each tablet containing 2800 IU of vitamin D is extracted into about 50 ml of about 5% water and about 95% methanol. Also in a specific embodiment, the resulting solution can be stirred for about 10 minutes, and ultrasonically shaken for about 30 minutes. Then stir for another 3 hours. In one embodiment, the sample can be separated by centrifugation at about 5,000 rpm 92852.doc -49- 200538136 to about 15,000 rpm. In a specific embodiment, the column is

Phenomenex Phenosphere 80 A ODS (l) column (150x4.6 mm, 3 μηι) with an injection volume of 100 μl. Dissolve the sample from the column and detect it. In a specific embodiment of the method, a 65-minute gradient can be used. A detection wavelength of about 260 nm to about 265 nm can also be used. In a specific embodiment of the method, the detection step is performed on a reverse HPLC column at about 25 ° C. The usable sample pan temperature is about 5 ° C. In a specific embodiment, the detecting step includes reverse HPLC separation using about 99% acetonitrile and about 1% 0.025% phosphoric acid. In a specific embodiment, the reverse HPLC column which can be used in the method of the present invention is not terminally capped or not terminally capped. Terminal capping method reduces the free silanol groups on the stationary phase, thus affecting the separation between pro-vitamin D and vitamin D waves. Attempts to use a terminal-capped column to provide sufficient peak resolution for the analytical method of the present invention have been unsuccessful because any dissolution of the degradation products between the two active ingredients cannot be resolved and resolved. In fact, when identifying a column for use in the method of the present invention, when the two active ingredients dissociated as seen everywhere, a vitamin ^ isomeric substance (O.:RRT) was observed. More development methods using other terminal capped columns are obvious. The analysis between vitamins and vitamin a waves is limited. :: The column carbon supports the product of transesterification of four pre-vitamin D3 or vitamin 仏 and medium chain triglycerides ((:: 8 and (: fatty acid two in combination). Invented the vitamin D sold by SF "Vitamin d3 has an effect. The purpose may be to react with vitamin H through the vinegarization mechanism to form CVD3 and Cl0 smoke. Because of the long fatty acid 92852.doc -50- 200538136 chain As a result, these esters are very hydrophobic and interact with the c18 stationary phase. A column with a higher carbon load has more c18 stationary phases; therefore, they interact more strongly with the esters and maintain the esters at Longer time on the column. Accordingly, in a specific embodiment of the method of the present invention, an HPLC column with a carbon load of less than about 10 ° / 0 can be used. A column with a lower carbon load can be used to reduce stationary phases and esters. These interactions caused these peaks to dissolve earlier. The results show that, for example, using a Platinum EPS C18 column with a low carbon load (5%), all esters are within 10 minutes when using 95% acetonitrile / 5% water in the mobile phase. Dissolve before. Similarly, when using another Phenosphere In the case of an ODS (l) column (7% carbon loading), all four esters were dissolved out within 26 minutes. The conditions of the chromatographic analysis exemplified in the method of the present invention are listed in the table below: : 1.2 ml / min column temperature: 25 ° C injection volume: 100 μl mobile phase: gradient, Α = 0.025% structural acid, B = 99% acetonitrile / 1% A running time: 65 minutes column: Phenosphere 80 A, ODS (l) column, 150x4.6 mm, 3 μm Sample plate temperature: 5 ° C Detector wavelength: 265 nm Gradient schedule: T (minutes) 0 16 39 43 57 57.01 65% aqueous solution 51.5 13 10 0 0 51.5 51.5% mixture 48.5 87 90 100 100 48.5 48.5 Using the detection method of the present invention, the pre-vitamin D and vitamin D peaks can be quantified to calculate the total amount of vitamin D in the sample. Specifically, the present invention The method is sensitive and selective. Samples with about 2800 IU of cholecalciferol can distinguish cholecalciferol, pre-cholecalciferol and its isomers, and detect one or more of 92552.doc -51-200538136 bile reduction Alcohol ester channels, or one or more pre-cholesterolized alcohol ester channels. In the stability study of the composition, three types of potential degradation products of vitamin D3 have been observed, which are described below (including Example 6). In the stability study described below, the bond studied The agent composition includes about 91.4 mg of sodium alentoric acid, about 26.7 mg of bilechol particles, about 11.0 mg of microcrystalline cellulose, about 62.4 mg of anhydrous lactose, and about 9.7 mg of cross-linked carboxylate. Sodium amidinocellulose, about 0.8 mg of colloidal silica and about 3.1 mg of stearic acid. As shown in Figure 5, the structure of vitamin A includes conjugated trienes that undergo various thermal and photochemical isomerizations. In the pharmaceutical composition exemplified in the present invention, we have identified 5 isomers of vitamin D3 (in this example, vitamin 03 (cholcalciferol) is used)-just-vitamin D3, trans-vitamin and three others are 〇78 RRT, 0 ·% RRT and 1.09 RRT isomers (this is a method for measuring the retention time of compounds by high performance liquid chromatography (HPLC) as described below). The structures of some of these vitamin d3 isomers are shown in Figure 5. The structural conclusions are based on UV, MS and, in some cases, 2NMR spectroscopy. Know

隹 玍, 隹 玍 京 〇 and its isomer pre-vitamin D will be heat-exchanged by the transfer of pro-sigma (S1gmatro Pic) 1,7 · hydrogen. In vivo, pro-vitamin D is the direct precursor of vitamin D. Although its balance tends to be larger, the biotin D 'is greater. However, the two things form a balance at physiological temperatures. Since vitamin D and pre-vitamin 0 are considered to have the same physiological functions, the commonly used vitamin 0 analysis report will include both. This is consistent with USP; Ph.Eur • research monographs for products containing, for example, vitamin D3. According to the stability data obtained, no other isomers can be properly packaged. Fang 92852.doc -52- 200538136 25 ° C / 60% RH, when stored for 24 months, it can reach 10% by weight. Monitor. The most obvious degradation product appears to be a vitamin 03 ester formed by the transacetation reaction of vitamin D3 and medium chain triglycerol wax (MCT) contained in the vitamin DJ granules used in the composition of the present invention. The structure of some of these vitamin channels is also shown in Figure 5. The main species are equivalent to n-octanoic acid A) and n-decanoate. The provitamin channel can be generated by the reaction of previtamin D3 with triglycerides in vitamin D3 compounds, or by thermal conversion of vitamin d3 vinegar. Among the quantifiable degradation products, only c8 and. Vitamin D3S channels will increase any degree of applicability in stability studies. Payable stability data show that these species should not reach the ICH threshold of 10 weight / 0 in 24 months when stored below about 30% relative humidity and below 30% relative humidity. The specific examples of the composition and method of the invention, in any case, we do not expect that it will give us safety concerns. Further research has shown that 'after 24 months, the total degradation product of the composition of the present invention is less than about 5% when stored below about thief and below ⑽%. We also know that vitamin D is induced by free radical initiators or spontaneously undergoes auto-oxidation in the solid or solution phase to form a variety of products, some of which have been identified. In a specific embodiment of the composition of the present invention, a representative characteristic of vitamin D degradation (specifically vitamin d3 in this example) was confirmed when vitamin d3 was converted into an oil or non-crystalline solid and exposed to 2 (M (rc temperature, The oxidative damage of vitamin d3. Within a few hours, the analysis of Ηρχχ showed the long-term destruction of vitamin d 923.doc -53- 200538136 and the occurrence of various unresolvable degradation products showing extremely low ultraviolet (UV) Absorption. At longer exposure times, these absorptions continue to decrease as further reactions occur. A more detailed analysis of vitamin D3 auto-oxidation utilizes the radical initiator nitrogen-bis-isobutylnitrile (AIBN ). In this experiment, the self-oxidation of vitamin D3 in the AIB_ ^ starting solution. The product was generated using UV, mass spectrometry (MS), and evaporative light scattering (EL_detection, determined by sail C). The results show that: the self-oxidation of the rhenium solution phase will also cause multiple degradation products' (b) auto-oxidation will gradually destroy the UV coloring performance, resulting in significant loss of the original 2; however, on the other hand, ELS detection provides significantly more Mass recovery results' (C) Mass spectrum m / z ratio and observed UV / visible spectrum of some cases confirm the oxidative characteristics of these reaction products. Radiometric calibration studies were performed to include about 7 () mg of alendurate And, the spoon of 2800 IU (70M grams) of vitamins used in the specific embodiment of the composition of the present invention, the granular formulation of vitamin h degradation characteristics. The use of standard vitamin D3 as tracking vitamin a degradation Method, which is independent of the change of the receiving characteristics. The radiolabeled vitamin A is incorporated into the vitamin D granules used in the specific examples of the composition of the present invention as a model prescription, and then its stability is analyzed. The amount of antioxidants is a reduced amount taken into consideration by commercially available prescriptions to ensure that degradation occurs within a reasonable time. After 14 weeks, liquid scintillation counting (LSC) and reversed-phase high-performance liquid chromatography ( RP-HPLC) plus 1; ¥ and online radiation detection, analysis of samples stored at 40 ° C / 75% RH and 70t. It can be observed that vitamin a stored at 40 ×: / 75% RH conditions is about 40% loss Among them, the low temperature control shows the better stability of 92852.doc -54- 200538136. The results of this analysis are listed in the radiochromatogram of the degradation product in Figure 6, which shows a large area of unresolved degradation products, none of which is shown as The main products. These results provide further evidence that when not properly stabilized, vitamin D3 is oxidatively degraded to a variety of products with reduced uV absorption, and these products explain the loss of vitamin 03. According to these stability analyses We do not expect individual oxidative degradation products to achieve safety concerns in specific embodiments of the bonding agent of the composition of the present invention. The present invention also includes a method for determining the pharmacological parameters of mammals when the composition of the present invention is administered. The pharmacological parameters can determine, for example, total urine excretion, urine excretion, and area under the plasma concentration versus time curve (AUC), Median Plasma Concentration (Cmax), Cmax Time (Tmax), and Plasma Concentrations in Lozenges such as, for example, Lozenges including about 70 mg alendurate and about 2800 IU cholecalciferol Obvious half-life Gw). These measurements confirm that specific examples of the compositions and methods of the present invention produce a pharmaceutically effective amount of alendurate and cholecalciferol in the body (the latter being the recommended daily dose of vitamin D through use with the compositions and methods of the present invention). Compare shown). In a specific embodiment, the present invention includes a method for measuring bileol in human blood aggregates after administering a pharmaceutical composition comprising alendurate and bile bilesol, the method comprising: (1) administering to a human A composition comprising alendurate and cholecalciferol, (2) obtaining a plasma sample from a human, (3) extracting bile from the plasma sample to form the first solution, and order the first Cholesterol in solution reacts with dienophiles to form one or more diels-alderM products of cholecalciferol. (5) Separation of cholecalciferol by high performance liquid chromatography (HpLC) separation method 92852.doc 200538136 Diels-alder addition product, and (6) using a mass spectrometer to detect the position of cholecalciferol in the sample. In a specific embodiment of this method, the dienophile includes 4-phenyl_1,2,4_triazoline-3,5-dione (P-TAD0 or PTAD). Moreover, the detection step can be performed in a positive ionization reaction mode using a heated cloud-like probe, and can further include adding deuterated internal standard cholecalciferol to each human plasma sample, and smoothing the sample bile. Calcified alcohol extracts, reacts, separates, and detects the deuterated internal standard cholecalciferol. When measuring 1 ml of plasma, this method has a quantitative limit (LOQ) of cholecalciferol of less than about 0.5 nanograms / ml of cholecalciferol. A specific embodiment of the present invention is a vitamin D / bisphosphate composition, in which the serum volatility map of mammals for 120 hours after applying the hair composition produces at least one of the following situations: the least square of cholecalciferol (LS ) The average AUCwum, hour> is about 296.4 nanograms · hour / ml. The determination of the pharmacokinetic parameters did not take into account the baseline reduction of blood cholesterol / month / agronomy 'minimum square (LS) average AUC ((M20 Hour) is about 297.5 nanograms · hour / ml, where the pharmacokinetic parameters, using the serum cholecalciferol concentration 0 hours before the dose as a covariate, taking into account the baseline cholecalciferol serum concentration; and The least squares (LS) mean Emei, hour) is about 143 nanometers · hour / ml, where the pharmacokinetic parameters are determined by subtracting the estimated baseline cholecalciferol after 120 hours, And consider the baseline _ alcohol concentration. In another specific embodiment, the group: the substance comprises a dibasic acid salt and fine alcohol, wherein a mammalian blood concentration map of the composition over 120 hours after administration of the composition produces at least the following-species: over 12 hours The mean value of the least squares (ls) of the steady state maximum blood concentration (c_) is about 5.9 nanograms per milliliter. Among them, the measurement of the pharmacokinetic parameters was not 92852.doc -56- 200538136 Considering the basis: bile feeding Serum concentration of alcohol; The average value of the minimum squared (LS) of the steady state of the hour is about 5.9 nanoliters. Among them, the determination of the pharmacokinetic parameters is based on the serum cholecalciferol concentration 0 hours before the dose. The covariates, 士, ^ spoon, and ali are measured taking into account the baseline of the serum of cholecalciferol; and by greedy energy-I, the minimum of the maximum blood concentration (Cmax) of shiwen ^ = average The value is about 4 phases of micrograms / ml, and when measuring the pharmacokinetics / count, the estimated baseline _ alkanol after the 12G hours has been subtracted, and the serum concentration of the baseline cholcalciferol is taken into account. The present invention also encompasses the composition. In i, the cholecalciferol and the blood of the cholecalciferol after 120 hours of administration of the composition have been shown to have a blood %% noise. Degree (cmax) is calculated as the average time of occurrence of cmax (Tmax), 12 hours, and ^ 'where the pharmacokinetic parameter does not take into account the baseline cholesteryl alcohol serum concentration. In a further example of your step back, the composition is calcified in the neck of a mammalian composition, Calcium & Liu Yue 7? The apparent median plasma concentration half-life is about 23.8 hours ; And 'The pharmacokinetic parameters are based on the baseline cholinesterol serum subtraction, taking into account the estimated baseline _ alcohol program. In order to determine the pharmacokinetic characteristics of the composition of the present invention, we conducted an open-label, crossover study of 236 healthy non-pregnant women and men between 18 and 65 years old. In this study, it is detailed in the following Example 7 'Vitamin a administered as 7 () mg of alendurate / 2 melons vitamin D3 combined tablets versus 2800 m of vitamin h tablets Pharmacokinetic parameters (AUCVm hours, Cmax, τ_ and median serum concentration half-month half-life (t ^)) were studied. In addition, the study of the combination bond relative to 92852.doc -57- 200538136 urinary excretion of alendurate salt of 70 mg fosamax tincture once a week. In short, (1) it has been shown that, with regard to the bioavailability of alendurate, 70 mg of alendurate / 280 mg of vitamins according to the present invention is used in combination with 70 mg Alendurate bond has the same biological equivalent, (2) 70 mg alendurate / 280m vitamin 仏 combination lozenge, and a lozenge containing 2800 IU vitamin D3 (without alendurate) The bioavailability of vitamin a is similar. (3) It has been shown that 70 mg of alendurate / 2800 IU of vitamin 03 combined tablets according to the present invention is very tolerable. Based on this, we expect that, for example, a bisphosphonate / vitamin D compound of the present invention administered once a weekly dose will provide the equivalent of a bisphosphate / vitamin D compound from a dose such as 〇 The recommended daily dose of vitamin D for vitamin D is the blood content and / or efficacy of vitamin Ds. These and other specific examples of this month will be further illustrated by the following non-limiting examples. Examples The following examples further illustrate and show specific embodiments within the scope of the present invention. _ Examples are given for illustrative purposes only and should not be construed as limiting the invention, as many changes may be made without departing from the scope and spirit of the invention. Example 1 The finished pharmaceutical product of bisphosphate and vitamin D lozenges contains sodium alendurate (about 70 mg of anhydrous free acid equivalent) and vitamin 03 (about 2_Lu • (about 70 micrograms)), Add the components identified in Table m ·. All formulations are rough and selected for maximum physical and chemical stability. 92852.doc -58- 200538136 Table 1-1 Ingredients Sodium alendurate 70 mg / vitamin D32800 I.U. Lozenges mg / bond weight% Sodium alentrum 91.37 28.1% Dry Vitamin D3100 granules 26.67 8.2% Microcrystalline Cellulose NF 131.0 40.3% anhydrous lactose NF 62.35 19.2% croscarmellose sodium NF 9.740 3.0% colloidal silica NF 0.8120 0.25% magnesium stearate NF 3.0870 0.95% total 325 100% used according to the method of the present invention The resulting lozenges are used, for example, to prevent, suppress, reduce or treat osteoporosis. Similarly, preparations of alendurate including other related weights of the base of alenduronic acid are prepared, including but not limited to approximately 2.5 mg, 5 mg, 8.75 mg, 17.5 mg, 70 mg, 140 mg per tablet Mg, 280 mg, 560 mg, or 1120 mg. Similarly, tablets containing vitamin D3 of other relevant weight per unit dose are prepared, including but not limited to approximately 1,400 IU, 2,800 IU, 5,600 IU, 7,000 IU, 8,400 IU, 14,000 IU, 28,000 IU per tablet Or 3 6,000 IU. These lozenges can be administered at intervals from once a week to once every two months. Example 2 Bisphosphate and Vitamin D Compositions Compositions including bisphosphate and vitamin D can be prepared using mixing and prescription techniques as described in this patent specification. A composition comprising about 35 mg of alendurate based on alenduric acid activity and about 5600 IU of vitamin D3 can be prepared using the following relative weight ingredients. 92852.doc -59 · 200538136 Ingredients Each bond Alenduric acid monosodium salt trihydrate 45.68 mg Dry Vitamin D3100 granules 56 mg * Anhydrous lactose, NF 71.32 mg microcrystalline cellulose, NF 80.0 mg magnesium stearate, NF 1.0 mg croscarmellose sodium, NF 2.0 mg * contains about 100,000 IU per gram of granules, so 56 mg of granules is equivalent to about 5600 IU. The resulting dosage form is used according to the method of the present invention, for example to prevent, suppress, reduce or treat osteoporosis. Similarly, alenduronic acid dosage forms based on alenduric acid activity basis including other relevant weights are prepared, including but not limited to about 2.5 mg, 5 mg, 8.75 mg, 17.5 mg, 70 mg, 140 mg per lozenge , 280 mg, 560 mg, or 1120 mg. Similarly, preparations of each unit dose including other relevant weights of vitamin D3, including but not limited to, each of the dosage forms are about 1,400 IU, 2,800 IU, 5,600 IU, 7,000 IU, 8,400 IU, 14,000 IU, 28,000 IU, or 36,000 IU ° These dosage forms can be administered from weekly to bimonthly. These dosage forms may be, for example, lozenges or capsules. Example 3 Alendurate and Vitamin D Lozenges Using the method disclosed herein, the following relative weight ingredients were used to prepare approximately 70 mg of alendurate based on alenduric acid activity and 2800 IU of vitamins. D3 tablets. 92852.doc -60- 200538136 Table 3-1 Composition (per lozenge) Sodium alendurate colloidal silica, CAB-0-SILP Dry Vitamin D3100 granules Microcrystalline cellulose NF Avicel PH-102 anhydrous lactose NF croscarmellose sodium rough grade magnesium stearate NF (non-fetal bovine) 91.37 mg t 0.81 mg 26.67 mg * 131 mg 63.35 mg 9.74 mg 3.09 mg t is equivalent to 70 · 0 mg free state acid * Dry VitaminD3100 granules also contain medium-chain triglycerides, gelatin, sucrose, medroxybenzylbenzene, starch and sodium aluminum silicate. * 26.67 grams of Dry Vitamin D3100 granules contain 105,000 IU / gram of vitamin D3. The produced lozenges are used according to the method of the present invention, for example, to prevent, suppress, reduce or treat osteoporosis. Similarly, tablets of alenduric acid based on other related weights of alenduric acid based actives are prepared, including but not limited to about 2.5 mg, 5 mg, 8.75 mg, 17.5 mg, 70 mg of each tablet. Home grams, 140¾: grams, 280 mg, 560 mg, or 1120 mg. Similarly, tablets of each unit dose including other relevant weights of vitamin D3 are prepared, including, but not limited to, about 1,400 IU, 2,800 IU, 5,600 IU, 7,000 IU, 8,400 IU, 14,000 IU, 28,000 IU or 36,000 IU. Such bonding agents can be administered at intervals from once a week to once every two months. Example 4 The absorption effect of vitamin D3 (powder type) on alendurate To test the effect of vitamin D3 (cholcalciferol) administered in a single dose on powder form of alendurate, we studied the effects of 14 85-year-old healthy, non-pregnant women and men underwent a two-stage, crossover study. Patients were instructed to receive one 70 mg alenduronic acid tablet every 92852.doc -61-200538136. In one of these two stages', a single dose of 5600 IU of vitamin D3 was co-administered with alendurate tablets according to computer-generated patient allocation progress. When vitamin D3 is administered with alendurate, the vitamin 03 powder is flushed back in 60 ml of normal tap water and applied to the patient with alendurate tablets (Treatment A). This vitamin 03 bottle was moistened and filled three times with 60 ml of normal tap water, and then administered to the patient each time. Therefore, a total of 240 ml in volume of ordinary tap water was applied together with vitamin D3. When alendurate is administered alone, a total volume of 240 ml of normal tap water is administered with the dose (Treatment B). Separate at least 14 days of clean-up at each stage. Treatment diagrams and assignments are listed in Table 4-1. Treatment Diagram and Assignment 7 and Beyond ~ '-Phase 1 Phase 2 (N = 7) ANs 0002, 0004, 0005, 0008, 0009, 00Π, 0013 AB-2 (N = 7) ANs 0001, 0003 , 0006, 0007, 0010, 0012, 0014 BA Zhiluo A 70¾ grams of alendurate tablets plus 5 ^ IU of vitamin B treatment 70 = alendurate. The patient was isolated in the research unit the night before each treatment. After overnight fasting (except water), the patient was given relevant treatment. After the application, the patient was fasted until the dose was administered. Two hours later, a fixed diet was administered. The clinical supply information is listed in Table 4-2. The composition and analysis results of alendurate tablets and vitamin Da used in this study are listed in Tables 4_3 and 4_4. 92852.doc -62- 200538136 Table 4-2 Clinically available drug efficacy dosage form Alendurate Vitamin d3 70 mg 5600 IU Lozenge granules Table 4-3 Alendurate Lozenge Formula Characteristic composition (each lozenge) Aaron Sodium succinate salt anhydrous lactose NF microcrystalline cellulose NF Avicel 102 magnesium stearate powder NF croscarmellose sodium NF type A 91.37 mg t 113.38 mg 140.00 mg 1.75 mg 3.50 mg t equivalent 70.0 mg of free-state acid. Table 4-4 Vitamin D3 Granular Powder Prescription Compositions (per bottle): Dry Vitamin D3 Type 100 CWS / HP 51.96 mg t 卞 equivalent to 5600 IU. All doses are administered after overnight fasting ( Except water). A dose of 70 mg of alendurate tablet plus 240 ml of regular tap water. When administered to a patient at a dose of 5600 IU of vitamin D3 plus alendurate, we instructed the patient to give 70 mg of alendurate Tartrate tablets are resuspended and co-administered with a vitamin D3 dose (provided in pellet form and flushed back with water at the study site). The total liquid volume per dose with alendurate dose was 240 ml. In the administration study After the drug, the patient was allowed to continue fasting for 2 hours, and a quantitative diet was given thereafter. The patient was allowed to stand up to 2 hours between the administration of the study drug and the quantitative diet. Each dose was separated at least 14 days apart 92852.doc -63- 200538136 Urine samples collected for alendurate analysis were collected for pharmacokinetic analysis at the following intervals ...-2-0 hours before dose, 0-8 hours after dose, 8-24 Post-hour dose and 24-36 hour post-dose. Urine collection during the 2 hour period before study drug administration provides the alendurate salt that we used to determine the baseline. 0-8, 8-24 and For 24-36 hours of urine collection, at intervals At the beginning, add 12-5 grams of boric acid to the container as a preservative. At the end of each time collection interval, the entire urine collection is weighed, the specific gravity is determined and the net volume is determined. The urine sample is acidified in place. Add 6.0 ml of hydrochloric acid (HC1) at a concentration of 6.0 equivalents per 200 ml of urine to make ρΗ ^ 2 · 0 of the urine sample. After acidification, agitate the urine sample and drop an equal amount of the sample into the polymer. The propylene container was stored frozen (-20 ° C) until high performance liquid chromatography (HPLC) analysis was completed. The total urine volume at each stage including boric acid and HC1 is used to determine the total urine excretion of alendurate at regular intervals. Analytical methods for determining alendurate in human urine involve three different know-hows: (1) separation of analytes from the urine and the internal standard (palmiturate), (2) formation of strong fluorescent derivatives (3) HpLC separation and fluorescence detection to generate derivatives. By adding calcium carbonate and sodium hydroxide, alendurate and intrinsic standard and naturally occurring phosphates are co-precipitated from the urine. The clot separated by centrifugation was backflushed in 1 mole hydrogen hydrogen acid, and added to an anion-exchanged diethylamine (DEA) cartridge in an acetate buffer solution at pH 4. Alendurate was dissolved out of the DEA box by a solution of 0.20 mol sodium citrate and 0.20 mol sodium disodium phosphate (adjusted to pH 9). Alendurate was derived at room temperature in the presence of N-ethylfluorenyl-D-penicillamine using 2,3-naphthyldiethylacetate. This derivative was then added to a non-silicone based polymer column consisting of styrene ethyl copolymer 92852.doc -64- 200538136 and diethylbenzene copolymer. The mobile phase initially consisted of 85% 0.025 Molar concentration of sodium citrate, 025 Molar concentration of disodium hydrogen phosphate, (pH 6.95), and 15% acetonitrile, and the flow rate was 1 ml / min. Dissolved later in the urine, the product is removed by increasing the acetonitrile concentration to 50%. With a coefficient of variation <10%, the analysis is valid in human urine between 5 ng / ml and 125 ng / ml. In order to obtain a detection limit of nanograms per milliliter, a 5-ml urine sample is required. Total urinary excretion of alendurate at specific intervals (-2-0, 0-8, 8_24, 24_36 hours) is by multiplying the concentration of alendurate in the equivalent analysis by the total of the interval Urine volume (including boric acid and hydrogen acid). The comparison of total urinary excretion of 70 μg alenduric acid tablets plus 5600 I1 Lj vitamin VII with 70 mg alenturic acid alone tablets is based on a two-stage, cross-designed Variation analysis (ANOVA) mode. The ANOVA model contains factors for order, patient (order), stage, and treatment. , ', Excretion of heart urine is logarithmic deformed. The result of the normal test by the office of the ______ world, plus the residue pattern by the "Hai mode", did not show any assumptions that deviated from the ANOVA mode. To estimate the relative bioavailability of 70 mg alendurate tablets plus vitamins relative to 70 mg alendurate tablets alone, we calculated the total urine excretion gmr to 95% CI based on the t-distribution . In addition, true posterior probabilities of clinically important boundaries exceeding 0.50 are also calculated. One patient withdrew from the above analysis because the alendurate concentration in all three collection intervals (0-8, 8-24, and 24-36 hours) for this particular patient was lower than the quantification for both treatments limit. Because the arrangement in the treatment order is slightly uneven, 92852.doc -65- 200538136. Data obtained from this patient for which we reported the mean of the least squares of total urine excretion were excluded from the analysis. The least square average is obtained by the AN0VA model by inverse transformation. All P values are rounded to three decimal places before the report. The result of its report of 0.050 is considered statistically significant. Table 4 shows that when each patient ingested 70 mg of alendurate tablets plus vitamin D3 and 70 g of alendurate tablets, the total urine of alendurate was excreted. The summary statistics with GMR and the total urine excretion of alendurate corresponding to 95% CI are listed in Table '6. The geometric mean of the least squares of total urinary excretion of 70 mg of alendurate tablets plus 5600 μv of vitamin 03 is 183.61, and for 70 mg of alendurate tablets alone Is 157.97. The gmr and its 95% CI corresponding to 70 mg of alendurate + vitamin Ds are 1.16 (0.74, 1.83) relative to those of alendurate alone. The GMR may exceed the clinically important 0.50 line and the posterior probability is 0.999. Table 4-5 Individual total urine of alendurate 36 hours after a single dose of 70 mg alendurate tablet plus 5600 IU of vitamin D3 and 70 mg of alendurate alone Excretion (μg) 92852.doc -66- 200538136 70 mg alendurate plus 70 mg alendurate 5600 IU vitamin D3 194.22 68.02 311.32 111.11 367.40 290.62 291.47 310.48 181.42 113.40 127.41 68.64 494.92 169.86 &lt; LOQt &lt; LOQt 21.75 61.29 185.46 257.75 101.51 68.73 97.86 259.69 248.71 341.90 464.51 519.58 237.54 203.16 The arithmetic mean standard deviation 143.63 140.51 t &lt; LOQ = Quantitative limit below 1 nanogram / ml Table 4-6 A single dose of 70 mg plus 5600 IU of vitamin D3 With individual administration of 70 mg of alendurate alone, individual total urine of alendurate over 36 hours: diarrhea (micrograms), summary statistics and geometric mean ratio of 95% CI for N LSt Mean Median Minimum Maximum SDi GMR§ 95% of GMR Cl11 Alendurate + 13 183.61 194.22 21.75 494.92 260.40 1. 16 (0.74, 1.83) Vitamin d3 Alendurate 13 157.97 169.86 61.29 519.58 177.07 Root Mean Square Error tLS Mean from Logarithmic Round Method of ANOVA Mode = Least Square Mean (inversely converted from Log Round Method) * SD = Inter-patient Standard The inverse conversion of the difference SGMR = least square mean ratio (alantoate + vitamin 03 / alantoate) CI = confidence interval 92852.doc -67- 200538136 Example 5 Vitamin D3 (included in alenduric acid Effect of salt / vitamin D tablets) on alendurate absorption

To test the possibility of the interaction of alendurate with orally administered vitamin D3, we administered 14 healthy adult subjects (6 males, 8 females, 33-61 years old) in a single-dose of 70 mg of alentol Acid salt, without vitamin D3, plus vitamin D3 (5 pairs of powder doses suspended in 240 ml of water. This study is a two-way design of openness, opportunity and intersection. The purpose of this study was to obtain vitamin D3 -Preliminary estimate of the relative bioavailability of alendurate after administration of 7G mg of alendurate lozenges relative to those who did not contain vitamin A.

/ In each of these two stages, the aluminate was orally administered in 7 () mg lozenges. In one stage, the lozenge is used with vitamin DA powder flushed in tap water &amp; in the replacement stage, the money is taken alone with tap water. For the analysis of excreted Asian salt, the urine was collected for 2 hours before each dose of alendurate, and 3 "hours thereafter. The relative biological benefit was based on the time after the dose was given. The total urine recovery of alendurate was evaluated. Urine recovered from 70 mg of alendurate without vitamin D3 was 202 micrograms, while 90% α was (126 micrograms, μ Micrograms); The average recovery of a 70 mg dose administered with vitamin d3- is 238 micrograms, and at the same time 同时 micrograms, 316 micrograms. The geometric mean ratio (α) is estimated to be 1.18 (0.80, 1.74). The Investigations have shown that orally administered vitamin D3 plus σ dose of sodium alendurate has the lowest to no effect on the bioavailability of alendurate 92852.doc -68 · 200538136. Example 6 Vitamin D3 and Aaron Stability Study of Tartrate We studied the stability of the composition of the present invention comprising a combined tablet form of sodium alendurate (70 mg of free free acid equivalent in anhydrous form) and vitamin D3 (2800 IU / 70 micrograms) Table 6-1 contains Alendurate / Vitamin Biotin D combined with lozenges in a specific embodiment of the lozenge composition. All excipients are rough grades, and their selection is to achieve maximum physical and chemical stability. _ Table 6-1 Lozenge composition Alenduric acid sodium salt 70 mg / vitamin D3, 2800 IU Lozenge mg / bond weight% Alenturonic acid 91.37 28.1% Dry Vitamin D3100 granules 26.671 8.2% microcrystalline cellulose NF 131.0 40.3% anhydrous lactose NF 62.35 19.2 % Croscarmellose sodium NF 9.740 3.0% Colloidal silica NF 0.8120 0.25% Magnesium stearate NF (intragranular) 2.275 0.7% Magnesium stearate NF (extragranular) 0.8120 0.25% Total 325 100% 92852.doc -69- 1 26.67 grams of Dry Vitamin D3100 granules contain 105,000 IU / gram of vitamin D3. Alendurate analysis and dissolution method can be used similar to the previously reported Fushanmei tincture tablets plus the front column Reversed-phase HPLC of 9-fluorenylmethyl chloride formate (FMOC) derivatives. The method of analysis and degradation of vitamin D3 can also be used to resolve and quantify the various phases of vitamin D3 and vitamin D3, which may reduce the reversed phase of the product. Gradient HPLC method (RP-HPLC). Vitamin D3 content consistency and dissolution analysis also uses reversed-phase HPLC. Because vitamin D3 is poorly soluble in water ’, this dissolution method can use a surfactant medium (1% SDS). Due to the low potency of vitamin D3 in the combined lozenge (70 micrograms), we can use 3 lozenges to 500 ml of vehicle to get the signal. The 52-week analysis and degradation product information for the entire batch of combined bonding agents stored at 30 ° C / 65% RH and 40 ° C / 75% RH are provided below (see Tables 6-2 and 6-3). Although the data produced do indicate a slight degradation of vitamin D3, these data do not show acceptable stability for the specific examples of the compositions of the present invention. Larger degradation was observed in higher temperature Ming foil vacuum formed packaging and HDPE bottles without desiccant. Table 6-2 Summary of the stability analysis results of Vitamin D3 Weeks of storage conditions t Vitamin d3 &lt; Aluminum foil sealed! Gram dry, M (% label statement) HDPE bottle of 75 ml J gasket, 1 good, 4 per bottle Tablets Vitamin D3 (% labeling aluminum foil vacuum forming packaging batch number 001 batch number 002 batch number 003 batch number 001 batch number 002 003 start 0 ^ 98.6 97.3 99.2 98.6 97.3 25 ° C / 60% RH ~ Ϊ ~ NT 97.3 99.5 99.2 97.7 26 99.9 98.0 NT 100.8 NT 39 99.0 NT 98.1 NT 93.8 44 99.1 97.1 99.6 97.9 95.4 _99T ^ 52 98.2 96.4 99.3 98.8 94.9 993 ^ 30〇C / 65% RH 13 ---- 100.1 96.8 NT 99.3 NT ^ 99T ^ 26 99.7 NT 99.5 NT 96.7 39 NT 94.4 97.1 97.6 94.3 NT ^ 44 97.3 95.7 97.6 97.9 96.1 H98O ^ 52 97.5 95.4 97.5 97.7 94.1 971 ^ 40 ° C / 75% RH ~ 13 99.3 96.0 99.0 97.1 96.1 ~ 26 97.1 94.5 96.9 96.8 94.7 The theoretical point in time-one NT = not tested 92852.doc -70- 200538136 Table 6-3 Summary of the stability results of vitamin D3 degradation: sodium alendurate 70 mg / vitamin D3 2800 1.U. Combination tablets, vacuum aluminum foil Storage packaging weeks of degradation products Statement% (% by weight relative to vitamin D) Lot No. 0001 Lot No. 002 Lot No. 003 0.74 RRT (trans-vitamin d3) Start 0 0.4 0.3 0.3 25 ° C / 60% RH 13 0.2 0.3 NT 26 0.2 NT 0.2 39 NT 0.3 0.2 44 0.2 0.3 0.2 52 0.2 0.3 0.2 30 ° C / 65% RH 13 0.2 NT 0.3 26 NT 0.3 0.2 39 0.2 0.3 NT 44 0.2 0.3 0.2 52 0.2 0.3 0.2 40 ° C / 75% RH 13 0.2 0.3 0.2 26 0.1 0.2 0.1 0 7 RRRT (vitamin D3 isomer) starting 0 0 · 〇ί o.ot o.ot 25〇C / 60% RH 13 0 · 〇ί 0.1 NT 26 0.1 NT 0.1 39 NT 0.1 0.1 44 0.1 0.1 0.1 52 0.1 0.1 0.1 30 ° C / 65% RH 13 0.1 NT 0.1 26 NT 0.1 0.1 39 0.2 0.2 NT 44 0.2 0.2 0.2 52 0.2 0.2 0.2 40 ° C / 75% RH 13 0.2 0.2 0.2 26 0.3 0.3 0.3 0.96 RRT (Vitamin D3 isomer) Initial 0 0.2 0.3 0.2 25 ° C / 60% RH 13 0.2 0.2 NT 26 0.2 NT 0.1 39 NT 0.2 0.1 44 0.1 0.2 0.1 52 0.1 0.2 0.1 30 ° C / 65% RH 13 0.2 NT 0.2 26 NT 0.2 0.1 39 0.1 0.2 NT 44 0.1 0.2 0.1 52 0.1 0.2 0,1 40 ° C / 75% RH 13 0.1 0.2 0.1 26 0.0 ^ 0 · 〇ϊ o.ot 1.09RRT (degradation of vitamin D3) Initial 0 NE NR NR 25〇C / 6 0% RH 13 ο.σ 0.〇i NT 26 ο.σ NT 0 · 0 this 39 NT 0_0 this 0.0 0.0 44 0.0; 0 · 〇ί Ο.Ο 92852.doc -71-200538136 Degraded matter storage condition weeks t Identification statement ° / 〇 (% by weight relative to vitamin D) Lot No. 001 Lot No. 002 Lot No. 003 52 o.ot o.ot o.ot 30〇C / 65% RH 13 o.ot NT o.ot 26 NT 0 · 〇 ί O.Oi 39 o.ot o.ot NT 44 o.ot 0.1 0.〇i 52 0.1 0.1 0.1 40 ° C / 75% RH 13 o.ot 0.0i o.ot 26 0.2 0.2 0.1 1 ^ QPPT (C8 Vitamin D3 ester) Starting 0 0.1 0.1 0.1 25 ° C / 60% RH 13 0.2 0.1 NT 26 0.2 NT 0.2 39 NT 0.2 0.2 44 0.3 0.2 0.3 52 0.3 0.3 0.3 30 ° C / 65% RH 13 0.2 NT 0.2 26 NT 0.2 0.3 39 0.4 0.3 NT 44 0.4 0.4 0.4 52 0.5 0.4 0.5 40 ° C / 75% RH 13 0.3 0.3 0.3 26 0.6 0.5 0.6 1.52 RRT (CIO Vitamin D3 Ester) Starting 0 o.ot 0.〇i 0.〇 i 25〇C / 60% RH 13 o.ot 0.0 NT NT 26 0.1 NT 0.1 39 NT 0.1 0.1 44 0.2 0.1 0.2 52 0.2 0.2 0.2 30 ° C / 65% RH 13 o.ot NT 0.1 26 NT 0.1 0.2 39 0.2 0.2 NT 44 0.2 0.2 0.2 52 0.3 0.2 0.3 40 ° C / 75% RH 13 0.2 0.2 0.2 26 0.4 0.3 0.4 Total degradation starting 0 0 .7 0.7 0.6 25 ° C / 60% RH 13 0.6 0.8 NT 26 0.8 NT 0.8 39 NT 1.0 0.9 44 0.9 1.0 0.9 52 1.0 1.1 1.0 30 ° C / 65% RH 13 0.7 NT 0.8 26 NT 1.0 0.9 39 1.0 1.1 NT 44 1.1 1.3 1.1 52 1.3 1.4 1.4 40 ° C / 75% RH 13 0.9 1.1 0.9 26 1.6 1.6 1.5 92852.doc -72- 200538136 Weeks of storage conditions of degradable substances 卞 Labeling statement% (wt% vs. vitamin D) Lot number 001 Batch No. 002 Batch No. 003 indicates the number of weeks at the theoretical point in time. Representative results &lt; 0.1% or untested NT = not tested NR = unreported stability results of vitamin D3 degradation summary: sodium alendurate 70 mg / vitamin D3 2800 IU Combined tablets, 75 cc HDPE bottles, 4 tablets per bottle and one gram of desiccant degradant, storage conditions, number of weeks t labeling statement (% by weight relative to vitamin D) Lot No. 001 Lot No. 002 Lot No. 003 0.74 RRT (trans -Vitamin d3) Start 0 0.4 0.3 0.3 25 ° C / 60% RH 13 NT 0.4 0.3 26 0.2 0.3 NT 39 0.2 NT 0.2 44 0.2 0.3 0.2 52 0.2 0.3 0.2 30 ° C / 65% RH 13 0.2 0.3 NT 26 0.2 NT 0.2 39 NT 0.3 0.2 44 0.2 0.3 0.2 52 0.2 0.3 0.2 40 ° C / 75% RH 13 0.2 0.3 0.2 2 6 0.2 0.2 0.2 0 78 RRT (vitamin D3 isomer) Starting 0 0.1 0 · 0 o.ot 25 ° C / 60% RH 13 NT 0.0 o.ot 26 0.1 0.1 NT 39 0.1 NT 0.1 44 0.1 0.1 0.1 52 0.1 0.1 0.1 30 ° C / 65% RH 13 0.1 0.1 NT 26 0.1 NT 0.1 39 NT 0.1 0.1 44 0.2 0.1 0.1 52 0.2 0.2 0.2 40 ° C / 75% RH 13 0.1 0.1 0.1 26 0.2 0.2 0.2 0.96 RRT (Vitamin D3 isomer) Initial 0 0.2 0.3 0.2 25 ° C / 60% RH 13 NT 0.2 0.2 26 0.1 0.2 NT 39 0.1 NT 0.1 44 0.1 0.2 0.1 52 0.1 0.2 0.1 30 ° C / 65% RH 13 0.2 0.2 NT 26 0.1 NT 0.1 39 NT 0.2 0.1 44 0.1 0.2 0.1 92852.doc -73- 200538136 Degraded matter storage conditions Week t (weight i label statement ° / 〇ί% relative to vitamin D) Lot No. 001 Lot No. 002 Lot No. 003 52 0.1 0.2 0.1 40〇C / 75% RH 13 0.2 0.2 0.1 26 0.0 * 0.1 0.0i 1.09RRT (vitamin D3 degradation product) Start 0 NR NR NR 25〇C / 60% RH 13 NT NR NR 26 o.ot o.ot O .ot 39 o.ot NT 0 · 0 44 44 O.Oi 0.0 2 52 0.1 o.ot 0.1 30 0C / 65% RH 13 0 · 0 0.0 0.0 NT 26 0.1 NT o.ot 39 NT 0.1 0.1 44 0.2 0.1 0.1 52 0.1 0.1 0.2 40 ° C / 75% RH 13 0.1 0.1 NR 26 0.2 0.1 0.1 1.39 RRT (C8 Vitamin D3 Ester) Start 0 0.1 0.1 0.1 25 ° C / 60% RH 13 NT 0.1 0.1 26 0.2 0.2 NT 39 0.2 NT 0.2 44 0.2 0.2 0.2 52 0.2 0.2 0.3 0.3 30 C / 65% RH 13 0.1 0.1 NT 26 0.2 NT 0.2 39 NT 0.3 0.3 43 0.3 0.3 0.3 52 0.4 0.3 0.4 40 ° C / 75% RH 13 0.2 0.2 0.2 26 0.5 0.4 0.5 1.52 RRT (C10 Vitamin D3 Ester) Start 0 0.0 * 0 · 0 this 0 · 0 this 25 ° C / 60% RH 13 NT 0 · 0 this 0 · 0 this 26 0.0 * 0.0ί NT 39 0.1 NT 0.1 44 0.1 0.1 52 52 0.1 0.1 0.1 30 ° C / 65% RH 13 o.ot o.ot NT 26 0.1 NT 0.1 39 NT 0.1 0.2 44 0.2 0.2 0.2 52 0.2 0.2 0.2 40 ° C / 75% RH 13 0.1 0.1 0.1 26 0.3 0.2 0.3 Total degradation starting 0 0.7 0.7 0.6 25〇C / 60% RH 13 NT 0.7 0.6 26 0.6 0.8 NT 39 0.8 NT 0.8 44 1.0 0.9 0.8 92852.doc -74- 200538136

t stands for result &lt; 0.1% or not tested NT = not tested NR = ▲ report

Example 7 The pharmacokinetic effect of vitamin D3 and alendurate M 23 36th, 18_65 years old non-annoyed; 1 F pregnant women and men are openly labeled 2 random, 2 parts, 2 The study of the crossover of stages. The study was conducted in two parts (parts-and two), each consisting of a two-phase, alternating design. Each patient participates in only-part of the study (ie each patient participates only in the first or part of the study). In each part of the study, the patients entered the study sequentially; in each part of the study, during the treatment phase, the washing phase was at least 2 to 12 days. The first part included treatments A and B. As for the first The second part includes treatments A and C. The treatment includes the following: Treatment of a single dose of 70 μg of Alenterate / 2_m vitamin D3 in combination with a single dose according to the following table M, treatment B- according to the single-dose of the following table 7_2-dose mg of aristoloate Acid salt bond, treatment c_ A single dose of 2800m vitamin 03 tablets (containing a placebo excipient in place of alendurate) according to Table 7-4 below. Urine was collected in Part- 'two hours before the start of the administered dose at each stage and at / λ' to determine total urine excretion of alendurate. 4th You'_24, _1, 8-, and 92852.doc -75- 200538136 before each stage of the application dose, the selected number of times during the hour, 0 hour (just before the administration) and 12 hours after the administration of the dose Collect blood to determine serum vitamin d3. · In the first part of the study, 7G mg of alendurate was evaluated according to Table 7_3 below. 2800 IU of vitamin D3 combined tablets and 70 mg of alendurate tablets according to Table 7_2 below Bioequivalence of Lentoate. In the second part of the study, the serum pharmacokinetic effect of vitamin 03 after administration of 70 mg alendurate / 280m vitamin a combined lozenges and 2800 IU vitamin lozenges was evaluated (AUC0- 120 hours, Cmax) 〇 The 2800 IU of vitamins 〇3 lozenges_ Contains 2800 R; inactive excipients of vitamin a and alendurate / vitamin a combination lozenges. The primary pharmacokinetic parameters in the first part were total urine excretion of alendurate 0 36 hours after administration of the oral dose. Because the plasma concentration after oral administration is very low and difficult to detect, the determination of total urinary excretion of alendurate is consistent with previous studies of oral bioavailability characteristics of alendurate via urine excretion. The primary pharmacokinetic parameters in the second part are AUCG_12㈣ 时 # and Cmax. Blood was collected at each of the following stages to determine the serum vitamin D3 concentration: _24, _18, _12, _6 hours, 0 hours (just before drug administration) and 2, 3, 5, 7 after drug administration , 16, 24, 36, 48, 72, 96 and 120 hours. Serum vitamin A concentrations were collected 24 hours before administration to provide an indication of the behavior of vitamin A endogenous activity in the control loop for 24 hours. Because vitamin A is synthesized on the skin by exposure to ultraviolet light, the patient is kept in the research unit for the duration of the pharmacokinetic sampling period (for example, up to 144 hours, 92852.doc -76- 200538136), 24 hours before the dose M hours after the dose was reached), no direct exposure; in i studies, patients were required to wear sun protection clothing (SPF45) and limit exposure days ^, including the washout phase. It also restricts patients' intake of high-vitamin D3 content (such as baby fish, goatfish, scalefish, finfish, finfish, swordfish, oysters and sardines), and foods supplemented with vitamin D3 (such as some Tinctures, fortified milk and certain cheeses). One patient in the mother of injury P received a single oral dose of 70 mg of alendurate / 28011; a combination tablet of vitamin D3 and a single oral dose of 70 mg in a randomized, alternating manner. Alendurate. In the second part, each patient received a single oral dose of mg of alendurate / 2800 IU vitamins &amp; a combination tablet and a single oral dose of 2800 IU vitamins &amp; Lozenges. Dosage was performed after fasting (except water) 'at 2100 the night before the dose was given, and used with 24 ml of tap water. Instruct patients not to lie down and stand upright (at least 45 degrees, sit or stand) between the drug application and the fixed meal. "Fast the patient until the standard dose is given two hours after the dose. Alendurate / Vitamins &amp; The method of combining lozenges is the same as that of alendurate. The compositions administered in the study are listed in the following Tables 7-1 to 7-4: / Vitamin d3 Alenturonic acid, Vitamin D3 70 mg / 2800 IU 70 mg 2800 IU Lozenges Lozenges 92852.doc -77- 200538136 Table 7-2 70 mg Alenturonic acid salt bond prescription composition (each Lozenges): sodium alendurate 91.37 mg ί '--- ~-microcrystalline cellulose NF 140.0 mg anhydrous lactose NF 113.4 mg croscarmellose sodium NF 3.50 mg magnesium stearate NF 1.75 mg content Analysis (alantoic acid) ·· Average i 70.42 mg Range 66.5-73.5 mg Ding manufactured by Merck's manufacturing department to become 70¾ g of sodium alanthionate ~-+ Free in 70 mM anhydrous free acid Table 7 -3 70mg Alendurate / 2800 IU Vitamin D3 Zirconium Sword Test_Composition (per lozenge): _ Sodium alendurate 1.37 mg t ~ --- Dry Vitamin D3100 granules 26.67 mg present anhydrous lactose NF 62.32 mg § Microcrystalline cellulose NF 131.00 mg colloidal silica NF 0.812 mg croscarmellose sodium NF 9.740 mg magnesium stearate NF (intragranular) 2.275 mg magnesium stearate NF (extragranular) 0.812 mg Content analysis (alantoic acid): average 70.5 mg RSD 0.89 % Range 69.7-71.6 mg content analysis (vitamin D3): average 2742 IU RSD 1.59% range ____ 卞 equivalent to 70.0 mg anhydrous free acid 2643-2822 ”,,, Qin Ge Wan ,,, y eight ,, on / Ding, No, Knife, and Bajia are two princes μ, and 4 accepting, such as methyltoluene, palace powder, and sodium glutamate. Made according to 5 analysis; 1 analysis of gelatin, sugar, and Ding reached 105,000 IU · The gram diaper is used for aging. It is estimated that a certain amount is used and the ink fixer weighs 325 mg based on the vitamin D amount of 100,000 IU · / g. ___ ^ 仃 Saqi · To achieve the end of the project 92852.doc -78- 200538136 Table 7-4 2800 IU vitamin D3 lozenge prescriptions

97.68 mg 46.61 mg 5.34 mg 0.445 mg 1.336 mg Composition (per lozenge) ...

Dry Vitamin D3100 Granules Microcrystalline cellulose NF anhydrous lactose NF croscarmellose sodium NF colloidal silicon dioxide NF magnesium stearate NF content analysis (vitamin D3): average RSD range

2789 IU 2.7% 2660-2957 IU + Dry VitaminD3100 pcs / ¾¾ Contains medium chain triglyceride No. Benzene, starch and sodium aluminum silicate. Make adjustments based on analysis. Presumption 4. Quantitative analysis and analysis reached 105,000 IU / g. Based on the amount of vitamin D3 added to 100 000 lu · / g, the weight of the lozenges has been reduced. ______ 一 ____ Use the unadjusted vitamin d3 concentration (ct) to calculate the area under the time curve (AUComch, hour) of the serum concentration versus time (AUComch, hour) after administration of the dose from 0 to 120 hours after the administration of the dose by the trapezoid method. Samples with concentrations below the LOQ limit are assigned zero values for calculation purposes. The time between the maximum observed concentration (Cmax) and cmax (Tmax) is obtained by measuring the concentration of vitamin d3 in the serum and confirming the recording of the sample collection. Vitamin d3 concentration in serum was also measured in three different ways, two of which explained the baseline serum vitamin 03 concentration in the following way. AUC 0.120 hours, cma ^ Tmaj are calculated in the same way. The bioavailability of the 70 mg alendurate tablet / 2800 IU vitamin combination tablet relative to the 70 mg alendurate tablet alone is based on the use of the alendurate / vitamin 03 combination tablet The dose was estimated relative to the GMR of alenturate total urinary excretion of 70 mg alendurate tablets alone. The relative bioavailability of the 70 mg alendurate / 2800 IU vitamin A combination tablet relative to 92852.doc -79- 200538136 as early as the 2800 IU vitamin C AI AI jtϊ cp f king i 1J3, using AUC0 -120 hours and Cmax GMR are estimated. The pharmacokinetic effect of the 70 mg alendurate / 280 m vitamin 03 combined tablet with the 28 mg IU vitamin D3 tablet alone after the administration of vitamin D3 was compared using three different methods. In the first method, the pharmacokinetic effect of vitamin h in the serum concentration of endogenous vitamin h is compared after the two treatments. Using this method, the serum concentration of endogenous vitamin a in the administration of 70 mg alendurate / 2800 m vitamin A combination tablets and 2800 IU vitamin D3 tablets was compared using the ANOVA model suitable for a 2-stage, cross-design. The pharmacokinetic effect of a single dose of vitamin 03 (AUCO) 20 hours and Cmax. Appropriate conversions are used for the pharmacokinetic parameters (i.e., the logarithmic conversion of the order of AUCowo hours, Cmax, Tmax 'and the inverse of the surface). It also reports inverse conversion summary statistics and inference results. Test the correctness of the ANOVA model assumptions. The correctness assumption usually meets AUC0_120 hours and Cmax.

In order to estimate the bioavailability of this 70 mg alendurate / 2800 IU vitamin D3 combination tablet with respect to the vitamin D3 of 2800 IU vitamin D3 tablet, the AMR between 0-120 hours and the CMR GMR (70 mg sub Lentoate / 2800 iu vitamin D3 combination tablets / 2800 IU vitamin D3 tablets) based on 90% CI of the t-distribution; then, compared with the previous specific bioequivalence boundaries (0 · 80, ι · 25). Summary statistics of Tmax for vitamin D3 and comparison between treatments are also provided. As another way of determining pharmacokinetic parameters, consideration of vitamin D3 in the plasma before dosing has led in particular to the development of a pattern of observed changes in vitamin D3 concentration during the experimental period. This model allows us to reduce the contribution of the baseline dimension of 92852.doc -80-200538136 to the serum concentration of biotin, and allows us to estimate the pharmacological parameters caused only by oral administration of this substance. This model relies on the following assumptions: ⑴ When no exogenous vitamin 仏 is used, the background inertia changes as a function of time in a linear fashion (Ci = Ci + Cm · t, where Ci and Q are the cuts of a straight line Distance and slope 'and t is relative to the number of hours of dose administration); ⑺ endogenous and ingested pharmacokinetic behavior of vitamin D3 are independent of each other, and it is also reduced whether or not additional doses are taken (for the purposes of this study) For the treatment or non-therapeutic), the body treats endogenously available vitamin 03 in the same way, and the intake of vitamin D3 is treated in the same way even if the body contains different amounts of this compound before the dose is administered; (3) When the dose occurs under the pharmacokinetic mode observed by the administration of a similar exogenous compound (the drug product of most swords), the concentration situation is returned to the baseline (that is, the terminal period of the regression to the baseline is a logarithmic straight line). Based on these assumptions, using the non-linear optimization method of Generalized Reduced Gradient performed by Micr0sftft Excel (soloverRoutin), CfCi + Cm will be described by the least square minimization method. The t-type baseline and the two-segment model (refer to the following equations) are substituted into the individual Ct vs. t cases (_24 to i hour after dosing). This substitution results in a logarithm of the terminal phase that approaches the baseline. The behavior of a straight line. Then use the best fit coefficient for each case to insert the baseline value into the range of 0-120 hours after the administered dose, and the interpolated baseline is subtracted from each case. = Ci + Cmt + Ae 'kd (t &quot; t, ae) + Be'kc, (t't, ae)-(A + B) e ~ ka (t't, ag) which is relative to the time of dose administration 92852.doc -81- 200538136 ct = predicted concentration of vitamin d3 in serum Ci = predicted value of baseline at t = 0 cm = slope of predicted baseline. As for A, B, kd, kel and ka, they have the first order of absorption. Parameter of segmentation pattern 'and tlag is the individual absorption delay after oral administration of vitamin 03. The pharmacokinetic parameters (AUcv12 (H, hour, Cmax, Tmax) determined by the method were calculated in the same way as described using the first measurement method. Summary statistics of total urine excretion of alenturate over 36 hours They are listed in Tables 7-5 below. After a single dose, total urine excretion of 70 mg of alendurate / 2800 IU vitamin D3 combined with 70 mg of alendurate lozenge alone was excreted. The average LS was 197.5 and 191.9 micrograms. Alendurate (70 mg alendurate / 2800 IU vitamin D3 combination lozenges vs. 70 mg of falconate lozenges alone) The gmR of the total urine excretion and the corresponding 90¼ CI are 1.03 (0.91, 1.17). The 90% CI falls within the previously specified bioequivalence boundaries (0.80, 1.25). Table 7-5 A single dose of 70 mg Alendurate / 2800 IU vitamin D3 combination bond or 70 mg Alentorate alone has the corresponding alenduric acid

Salt (Winter) ^ 90% confidence zone! The total anti-liquid excretion is to be broadcasted rTlvfR treatment N LSt mean median minimum maximum SD SD GMR§ 90% CI of GMR Salt + Vitamin d3 Alendurate 207 207 197.5 191.9 204.4 11.3 0.1 3617.7 1629.6 329.1 522.2 1.03 (〇.915 1.17) By ANOVA mode tLS = lowest mean (Θ iSD = by logarithmic scale § GMR = geometric mean ratio 41 = The error mean square error of the confidence interval ten scale b log scale inverse conversion). ίConverted patient-to-patient standard; 1, value (average LS of alendurate + vitamin (RMSE) = 0.778 0--0 of D3 / average LS of alendurate). 92852.doc -82- 200538136 Relative to plasma determination, the mean LS of vitamin D3 (aucg_12CH, eight without considering the serum concentration of endogenous vitamin D3) vs. 70 mg alendurate / 2800 IU vitamin D3 combined tablets with 28〇IXJ vitamin D3 lozenges are 296 · 4 and 337.9 nanograms · hour / ml (Table 7-6). 0AUC0-120 hour GMR (alantoate plus vitamin D3 combined lozenge / vitamin d3 lozenge) It was 0.88 and 90% CI was (0.81, 0.95). Table 7-6 70 mg alendurate / 2800 IU vitamin D3 lozenges or 2800 IU vitamin D3 lozenges alone after administration of a single dose does not take into account the endogenous vitamin D3 serum concentration of aucg_12 with the corresponding vitamin 仏 ( H, hour (femto

&amp; · Small / ml) Summary statistics of 9 ^^ Lai interval and GMR treatment N LSt Mean median minimum maximum value SDt between patients GMR§ 90% of GMR Cl II Alendurate / Vitamin d3 Vitamin d3 alone 28 28 296.4 337.9 257.5 309.6 85.0 111.9 1648.8 1485.9 375.5 344.2 0.88 (0.81, 0.95) Root mean square error () t by logarithmic scale iSD = by logarithmic ^ §GMR = geometry i trust zone R EIMSE) = 0.168 (reverted by ANOVA; the least squared average of the inversion is only the standard deviation of the inverse conversion of the late mode. The average production ratio (vitamin D3 + Aaron · expression). The average LS of glutamate / the average LS of vitamin D3 &gt; straight) . When the serum concentration of endogenous vitamin D3 is not taken into account, the mean LS of vitamin E &gt; 3 Cmax for 70 mg alendurate / 2800 IU vitamin D3 combination tablets and 2800 IU vitamin D3 tablets is 5.9 and 6.6 milligrams, respectively. Μg / ml (Table 7-7). CmaxGMR (alantoate plus vitamin d3 combined lozenge / vitamin D3 lozenge) was 0.89, and its 90% CI was (0.84, 0.95). Regardless of the AUCo-uch of the serum concentration of vitamin 〇3, and the 90% CI of CmaxGMR fall within the previous specific biological equivalent range (0.80, 1.25). 92852.doc -83-200538136 Table 7-7 A single dose of 70 mg alendurate plus 2800 IU vitamin D3 combination sharps or 2800 IU vitamin D3 revolver alone does not take into account the serum concentration of endogenous vitamin D3 Cmax with corresponding vitamin D3 (femto

G / ml) at 90 ° /. Summary statistics of confidence interval and GMR treatment N LS + Mean median minimum value Maximum value SDt between patients GMR§ 90% of GMR CI 丨 Alendurate / vitamin d3 Vitamin d3 alone 28 28 5.9 6.6 5.3 6.2 2.5 3.5 17.4 18.1 3.3 3.1 0.89 (0.84, 0.95) Root mean square error (] t by logarithmic scale iSD = by logarithm ^ SGMR = geometry = trust region R SE) = 0.138 (by ANOVA mode). Least squares mean of inverse conversion. fe See standard deviation of inverse mode conversion. Yield-to-average ratio (average LS of vitamin D3 + alendurate / average LS of vitamin D3). Tables 7-8 show the results of statistical analysis of vitamin D3Tmax obtained from serum without considering the serum vitamin D3 serum concentration. The median Tmax of vitamin D3 with or without alendurate was 12.0 and 9.0 hours, respectively. No significant difference was observed between the treatments (p-value> 0.200). Table 7-8 A single dose of 70 mg alendurate plus 2800 IU vitamin D3 lozenges or 2800 IU vitamin D3 lozenges alone Vitamin D3Tmax derived from serum conditions without regard to endogenous vitamin D3 serum concentration Summary of p p t (hour) SDt p value between patients with median minimum and maximum median value Alendurate / vitamin d3 Vitamin Eh alone 28 28 12.0 9.0 7.0 7.0 16.0 16.0 2.6 2.3 0.978 TSD = standard deviation. ίρ-values are calculated using hierarchical analysis. For 70 mg alendurate / 2800 IU vitamin D3 combined tablets and 2800 IU vitamin D; tablets of vitamin D3 aUCg_12 (average of LS for h and hour 92852.doc -84- 200538136 respectively 297.5 and 336 7 nanograms / hour / ml (Table 7-9). AUC0-120 hours GMR (alendurate plus vitamin D3 combination lozenge / vitamin D3 lozenge) is 0.88, and its 90% CI is 82, 〇95). Table 7-9 A single dose of 70 mg alenduronate plus 2800 IU of vitamin D3 combined lozenges or 2800π vitamin D3 lozenges alone, with a vitamin D3 concentration of time = 0 before the lozenges were administered Summary of the variables with the corresponding vitamin DsAUCcm: ^, hour (nanogram · hour / ml) confidence interval of 90 / ()

Calculate GMR treatment N LSt Mean Median Minimum Maximum Maximum SDi GMR§ 90% of GMR CI11 Alendurate / vitamin d3 Vitamin d3 alone 28 28 297.5 336.7 257.5 309.6 85.0 111.9 1648.8 1485.9 376.8 343.0 0.88 (0.82, 0.95) Root mean square error (RMSE) = 0.154 (referred to by ANOVA as the minimum squared mean of reversal from logarithmic scale $ SD = standard deviation of inverse conversion from logarithmic scale. SGMR = geometric mean ratio (vitamin d3 + Aaron wants 41 = confidence interval; formula). 0 • LS average of acid salt / LS average of vitamin D3). As shown in Tables 7-10 below, the average LS of vitamin D3 for 70 mg alendurate / 2800 IU vitamin D3 combination tablets and 2800 IU vitamin D3 tablets is 5.9 and 6.6 nanograms / ml, respectively. (: _〇] ^ 11 (70 mg alendurate / 2800 IU vitamin D3 combination tablets / 2800 IU vitamin D3 tablets) is 0.90, and its 90% CI is (0.85, 0.95). Time = 0 The vitamin D3 concentration before the administration dose was a covariate of AUC0-120 hours and 90% CI of CmaxGMR fell within the previous specific bioequivalence range (0.80, 1.25). Table 7-10 A single dose of 70 mg alendurate Plus 2800 IU vitamins 92852.doc -85- 200538136

〇3 combination bond or separate 2800 IU vitamin D3 lozenges, the time of administration of lozenges 〇 = vitamin D3 concentration of 〇 is a covariate, summary statistics with 90% confidence interval of corresponding vitamins and GMR treatment N LSt average Alan female salt / vitamin d3 Vitamin d3 alone 28 28 5.9 6.6 5.3 6.2 Minimum value 2.5 3.5 Maximum value 17.4 18.1 Patient-to-patient SD * 3.3 3.1 GMR§ 0.90 90% of GMR Cl &quot; (0.85, 0.95) t The logarithmic scale inverse transformation is the mean of the squared '. tSD = standard deviation of inverse conversion on a log scale.

^ GMR = Geometric Mean Ratio (Vitamin D3 + LS Means of Alendurate / Vitamin D3 Means) 41 = Vitamin D3AUC0-12 (H, Time The results of the data analysis are summarized in Tables 7-11. 70 mg alendurate / 2800 IU vitamin D3 combination tablets and 2800 IU vitamin D3 tablets of vitamin D3 AUC0_120 hours were measured using the model-based vitamin D3 baseline concentration. The average LS was 143.1 and 169.1 ng · hr / ml. The AUC0-i 20-hour GMR (alentorate plus vitamin D3 combined lozenge / vitamin D3 lozenge) was 0.85, and its 90% CI was (〇 · 76, 0.94). The lower limit of 90% CI falls below the previous specified lower limit of 0.80. Table 7-11

Corresponding vitamin D3AUCg.12 (H, hour (nanogram · hour), as measured by a single dose of 70 mg alendurate / 2800 IU vitamin D3 combination lozenges based on the vitamin D3 baseline concentration based on the mode of use / Ml) summary statistics of 90% confidence interval and GMR 92852.doc -86- 200538136 treatment N LS 卞 mean median minimum maximum value SDi GMR between patients 90% of GMR Cl &quot; Salt / vitamin d3 Vitamin d3 alone 28 28 143.1 169.1 153.5 175.0 61.1 107.2 236.1 251.4 47.7 37.3 0.85 (0.76, 0.94) Root mean square error () 卞 by logarithmic scale by logarithm; §GMR = geometry j uci = trust zone p RJVISE ) = 0.224 (Lean squared mean value transformed by ANOVA's standard deviation of inverse conversion in late mode. ^ Mean ratio (Vitamin) 3 + Aaron 妾 0: LS mean of acid salt / LS of vitamin D3 (Mean value) 0 As shown in Table 7-12, 70 mg of alendurate / 2800 m vitamin d3 combination tablets and 2800 m of vitamin D3 tablets of vitamin D3 were used based on the model-based vitamin D3 baseline concentration. The average LS Cmax is 4.0 and 4.6 ng / Liters. CmaxGMR (70 mg alendurate / 2800 IU vitamin 03 combined tablets / 2800 IU vitamin d3 tablets) was 0.88, and its 90% CI was (0.83, 0.93). Utilization mode-based vitamins 90% CI of CmaxGMR as measured by A baseline concentration falls within the previous specific bioequivalent boundary (080, 1.25) ° Yi / _12 A single dose of 70 mg alendurate plus 2800m vitamin D3 combination Tablets or separate 2800 ⑴ vitamin A troche use mode based on vitamin D3 baseline concentration measured with corresponding vitamin treatment N 〇 LSt mean median minimum maximum SDI GMR between patients § ^ 90% of UMK GMR PT11 hypoepitate / vitamin d3 Vitamin D3 alone error root mean square () 28 28 EIMSE 4.0 4.6 1 &gt; 〇Λ \ 5 (FREE 4.0 4.6 ΑΝΟΛΜ lift 1.9 3.0 -4- '\ 6.0 7.2 1.1 0.9 0.88 7 V // 〇L 丄 (0.83, 0.93)-▼ 『-\ 一 JVAA w / I &gt; \ JV Μ ^ 平均值 The least square average value inversely converted from logarithmic scale: 矻 0 = Standard deviation of inverse conversion from a logarithmic scale ^ Mean ratio (average LS of vitamin alendurate / vitamin D3 LS mean value). — 1 ^ ~ -------—__ Table 7-13 shows the results of vitamin 仏: ^ statistical analysis of the situation obtained from the determination of the baseline vitamin h concentration based on the utilization mode 92852.doc 200538136. The median D3 of this vitamin with or without alendurate was 120 and 90 hours, respectively. No significant difference was observed between the treatments (13 values> 0 2 00). Table 7-13 Vitamin U3Tmax obtained from a single dose of 70 mg alendurate plus 2800 IU of vitamin D3 combined tablets or 2800 IU of vitamin A tablets alone based on the baseline vitamin concentration determination (Hours) Voice Statistics

Table 7-14 summarizes the results of the analysis of the vitamin concentration concentration based on the use model and quasi-biotin 03: 1 /: 2. With or without alendurate

Significant tw2 of the harmonic mean of vitamin 03 was 24.0 and 25.5 4 respectively. No significant difference between treatments was observed (p value> 0.200).

Table 7-14. Significant vitamins determined based on baseline vitamin Ds concentration based on the utilization pattern of a single dose of 70 mg alendurate / 2800 IU vitamin D3 combination tablet or 2800 IU vitamin A tablet alone. Heart 2 (hours) summary statistics

92852.doc -88- 200538136 Example 8 Degradation detection method A combination of alendurate / vitamin 03 bond (70 mg alendurate / 2800 IU vitamin DO of vitamin 03) has been developed. Synthetic analysis method. Vitamin D3 is extracted from 15 lozenges in about 50 ml of 5d / 0 water / 95% methanol diluent solution. The solution is stirred for 10 minutes, ultrasonically shaken for 30 minutes, and stirred for another 3 hours. Centrifuge the sample and inject 100 microliters of the supernatant into a Phenomenex Phenosphere 80 A DS (l) column (150x4.6mm, 3 microns) for reverse-phase HPLC analysis. The detection wavelength is 265 nm 65-minute gradient method. Pre-vitamin D3 and vitamin A peaks were quantified and totaled to calculate the total vitamin D3 in the sample. The method was validated to meet specificity, linearity, recovery, accuracy, and repeatability The stability, sensitivity, and strength of the alkaline solution are shown in the table below. Flow rate: 1.2 liters / min. Jiangli official column temperature: 25 ° C —-- 1, 100 μl ~~ --- Mobile phase: gradient, A = 0.025% phosphoric acid, B = 99% acetonitrile 7ϋ ~-running Duration: 65 minutes &quot; Column: Phenosphere80A, ODS (l) column, 150x4.6 mm, 3 micron sample tray temperature: 5 ° C Detector wavelength: 265 nm Gradient schedule: T (minutes) 0 16 39 43 57 57.01 65% aqueous solution 51.5 13 10 0 0 51.5 51.5 _% mixture 48.5 87 90 100 100 48.5 48.5 92852.doc -89- 200538136 combination lozenge composition (70 mg alendurate / 2800 IU Vitamin D3) Unit weight of the composition (mg) Weight% Sodium alendurate 91.5 28.2% Dry Vitamin D3100 granules 26.7 * 8.2% Avicel PH102 131 40.3% anhydrous lactose 62.2 19.1% croscarmellose sodium 9.75 3.00% colloid Silicon dioxide 0.81 0.25% intragranular magnesium stearate NF 2.28 0.70% extragranular magnesium stearate NF 0.81 0.25% lozenge weight: 325 100% * 26.7 grams of Dry Vitamin D3 100 granules containing 105,000 IU / g vitamin D3 Table 8-1 lists typical retention times and relative retention times (RRT) of excipient peaks and degradants relative to vitamin D3. As shown in Figure 5, the main degradation pathways of vitamin D3 are photoisomerization, thermal isomerization, and transesterification. Table 8-1 Summary of peak identification of combined tablets Retention time (minutes) RRT classification 3.08 0.09 related to excipients 3.63 0.10 related to excipients 3.88 0.108 related to excipients 4.05 0.11 related to excipients 4.23 0.12 related to excipients Related to vehicle 4.78 0.13 Related to vehicle 7.53 0.21 Unknown 11.33 0.32 Related to vehicle 17.25 0.48 Unknown 17.47 0.49 Related to vehicle 18.05 0.50 Related to vehicle 20.64 0.57 Related to vehicle 26.70 0.74 Trans-vitamin d3 28.15 0.78 Vitamin d3 isomer 92852.doc -90- 200538136 31.33 --- 0.87 Pre-vitamin d3-34.42 0.96 Vitamin D3 isomer-35.94 1.00 Vitamin d3 39.17 --- 1.09 Vitamin d3 isomer-43.45 1.21 and Excipients related 49.85 1.39 C8-vitamin D3 ester—50.42 1.40 C8-vitamin D3 ester_ 54.52 1.52 C10-vitamin D3 ester—55.65 1.55 C10 pre-vitamin 03 ester Determined by the main injection of different concentrations of vitamin D3 solution (10Q) and select the lowest concentration with a signal-to-noise ratio of 10 or more. The determined L0q ^ is about 9 nanograms / ml (100 microliters of injection volume), which is 0.04% of the method concentration with an average signal-to-noise ratio of 11.1 determined in 10 iterations. Example 9 Dosage therapy with one dose per week We can prepare alenduronic acid salt containing about 70 mg of altartoic acid salt based on alenduric acid activity and about 5,600,000 vitamin D of alendurate With vitamins £) lozenges, or other solid dose prescriptions (see examples 丨, 2 and 3). The lozenge or other solid dose prescription can be administered orally to a patient once a week, that is, once every 7 days with _, spoon, preferably (for example, every week) for a period of at least one year. We expect that the Haitian method will be useful and convenient for treating or preventing osteoporosis while providing vitamin D replacement. We also expect this method to be used to improve ϋacceptance and obedience ’and to ensure that all patients taking biquatate can receive adequate vitamin D nutrition. · 代 Generation 'we can prepare about 70 mg of almonduric acid activity ** ρ Chu hypophagenic acid salt, and about vitamin D alenduronic acid salt and biotin D tablets' or other Solid Dose Prescription (see Example 3 and 3). The tablets 92852.doc -91-200538136 or other solid dose formulations can be administered orally to patients once a week, that is, preferably once every 7 days (for example, every Sunday) for a period of at least one year. This method of administration is expected to be useful and convenient for the simultaneous treatment and prevention of osteoporosis and biotin D nutrition. We also expect this approach to be used to improve patient acceptance and compliance, and to ensure that all patients taking bisphosphonates receive adequate vitamin D nutrition.

Alternatively, we can prepare lorentulate containing about 35 mg to about 70 mg, based on the activity of esophageal acid, and hypochondrite and vitamin D tablets of about 2,8 G0IU vitamin D, or Other solid-dose prescriptions (see Example 31): The bond or other solid-dose prescriptions can be administered orally to a patient orally once a week, that is, once every 7 days (eg, every Sunday) for at least one month This method of application is expected to be useful and convenient for providing vitamin D nutrition for the treatment or prevention of osteoporosis. We also expect this method to be used to improve patient acceptance and compliance, and to ensure that all patients taking double-filling salts can receive adequate vitamin D nutrition.

We can prepare alenduric acid salt and vitamin D ^ 'or other solid dosage formulations containing about 280 mg of altartoic acid salt based on alenduric acid activity and about 5,600 IU of vitamin D ( See Examples 2 and 3). This rotatable or corpus callosum dosage formulation can be administered orally to a patient once a week, that is, ^ about once every 7 days is preferred (for example, every Sunday) for at least one to six months. The application method is useful and convenient for treating osteitis osteitis and providing vitamins and electrics. We also expect this method to be used to improve patient accessibility and compliance, and to ensure that all ingested bilinamic acid can receive adequate vitamin 0 nutrition. U

92852.doc -92- 200538136 generation, we can also prepare alenduronic acid salt containing about 280 mg based on alenduric acid activity, and about 2,800 IU of vitamin D alendurate and vitamin D lozenge, Or other solid dose formulations (see examples). The lozenges or other solid dose formulations can be administered orally to a patient once a week, preferably at about every 7 days (for example, every Sunday) for at least one to six We hope that this method of administration will be useful and convenient for the simultaneous treatment of teratogenic osteitis / vitamin D. It is also expected that this method can be used to improve patient acceptance and compliance and to ensure that all bisphosphonates are ingested Patients can receive sufficient nutrition of vitamin D. We can prepare about 280 mg of alendurate based on the activity of alenduric acid = lentulate, about 5,600 IU or edulinate D ... D lozenges 'or other solid dose prescriptions (see Example 2) Pastes or other solid dose prescriptions can be administered orally to patients once a week', that is, about every 7 days __ / a /, .. (For example, every Sunday). This method of administration is useful and convenient for treating metastatic osteochondrosis at the same time providing vitamins = nourishment. We also expect that this method can be used to improve: compliance and ensure that all patients taking double salt are: adequately The nutrition of vitamin D. In the case of -.w which invention or application is dependent on God, it is obvious that this composition can be used to inhibit bone loss, which is very obvious. We also intend to declare patents in 4 and the attached The scope covers the correction, changes, and equivalent methods of the prescription composition of the present invention that inhibits moon scallops. [Simplified Illustration] 92852.doc -93- 200538136 Figure 1 illustrates the metabolism of vitamin d3. Figure 2 illustrates cholecalciferol and The chemical structure of alendurate monosodium salt trihydrate. Figure 3 illustrates the chemical structures of vitamin D2 and vitamin D3. Figure 4 shows a schematic representation of a summary of a specific example of a method of preparing the composition of the present invention. Figure 5 illustrates Thermal and photochemical isomerization and transesterification of vitamin D3. Figure 6 shows the results of a radiometric study of vitamin D3 degradation. 92852.doc 94-

Claims (1)

200538136 10. Scope of patent application: 1. A pharmaceutical composition, which includes: a bisphosphonate, a salt, derivative, or hydrate of a bisphosphonate, or a mixture thereof, and a vitamin D compound. 2. The pharmaceutical composition according to claim 1, wherein the bisphosphonate has the formula: HO I «1 I OH I 0: 1 = P — 1 I — C — II — p 0 1 HO IL where Rl is independently selected Since Η, OH and Cl, R2 is independently selected from CH3, C1, CH2CH2NH2, (CH2) 3NH2, CH2-3-pyridyl, CH2-S-phenyl-C1, ch2ch2n (ch3) (pentyl), CH2- Weltazole, CH2-2-imidazole-romidinyl, N · (cycloheptyl), CH2ch (Ch3) 2, (CH2) 5NH2 and CH2-1-pyrrole, and combinations thereof. 3_ The pharmaceutical composition according to claim 1, wherein the bisphosphate comprises alendurate or a pharmaceutically acceptable salt thereof. 4. The pharmaceutical composition according to claim 3, wherein the medically acceptable salt of the alenduric acid salt is selected from the group consisting of alenturic acid monosodium salt, alenturic acid monosodium salt monohydrate, and alentoric acid Acid monosodium salt trihydrate. 5. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises a vitamin 0 compound from about 100 IU to about 36,000 11; 6. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises from about 0.5 mg to about 1120 mg of a bisphosphate, or a pharmaceutically acceptable salt, derivative, or hydrate of the bisphosphate, Or a mixture thereof. 92852.doc 200538136 7. The pharmaceutical composition according to% of claim 1, wherein the pharmaceutical composition comprises about 2'8000111 of cholecalciferol and about 70 mg of alendurate, or alendurate Pharmaceutically acceptable salts, derivatives or hydrates, or mixtures thereof. 8. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises about 5,600,001 of cholecalciferol and about 70 milligrams of alendurate, or a side effect of alendurol. Party salt, derivative or hydrate, or a mixture thereof. 9 The pharmaceutical composition according to claim 1, further comprising one or more excipients' which is selected from the group consisting of a filler, a diluent, an adhesive, a lubricant, a lubricant, and a decomposing agent. 1 10. The pharmaceutical composition according to claim 1, further comprising one or more excipients selected from the group consisting of anhydrous lactose, microcrystalline cellulose, colloidal silicon dioxide, sodium parent carboxymethylcellulose and hard A group of magnesium stearate. The pharmaceutical composition according to claim 10, wherein the pharmaceutical composition comprises about 4 to 90 /. The weight of sodium fumarate, from about 1% to about 20% by weight of the bile-reduced alcohol particles (equivalent to about 0.0005Q /% to about the weight of the bile-reduced alcohol) is about 10. /% To about 80% by weight of anhydrous lactose, about 5% to about 50% by weight of microcrystalline cellulose, about 0.01% to about 5% by weight of colloidal dioxide, and about 0.5%. To about 10% by weight of cross-linked methylcellulose sodium, and about 0.5% to about 5% by weight of magnesium stearate. 12. The pharmaceutical composition according to claim 7, wherein the Guanhua alcohol comprises a pharmaceutical grade bile # 5hua alcohol. 13. The pharmaceutical composition according to claim 12, wherein the pharmaceutical composition is suitable for administration intervals of once a week, once every two weeks, once a month, twice a month, and once every two months. 92852.doc 200538136 14. · A method of preparing an alendurate-cholesterol composition, comprising: preparing a mixed powder including alendurate, and compressing the mixed powder to form an alendurate mixture | ) Grinding and mixing the alendurate mixture with cholecalciferol particles to form a final mixture; and lubricating and compressing the final mixture. 15. —A method for preparing alendurate salt_cholesterol solid dosage form, which comprises: alendurate salt, colloidal silica, anhydrous lactose, microcrystalline cellulose and croscarmellose sodium Mix to form a premix, mix the premix with magnesium stearate to form the first lubricating mixture, compress the roller of the first lubricating mixture to form a compressed strip, and grind the compressed strip to form lubrication The second lubricating mixture is mixed with the cholesteryl alcohol particles to form a second lubricating mixture; and the second lubricating mixture is compressed into a solid dosage form. 16. A pharmaceutical composition manufactured according to the method of claim 15. 17. The pharmaceutical composition according to claim 1, which is prepared by a method comprising the steps of: incorporating about 0.5% to about 90% by weight of sodium alendurate, and about 0.1% to about 5% by weight of a colloid Silicon dioxide, about 10% to about 80% by weight of anhydrous lactose, about 5% to about 50% by weight of microcrystalline cellulose, and about 0.5% to about 10% by weight of croscarmellose fiber Sodium, mixed with about 5% to about 5% hard 92852.doc 200538136 magnesium stearate and other ingredients to form the first mixture. The first mixture roller is compressed to form a compressed strip. The bar is ground to form a lubricating mixture, and the lubricating mixture is mixed with about 1% to about 70% by weight of cholecalciferol particles (equivalent to about 0.0005% to about 20% by weight of cholecalciferol) to form a second This mixture is used to condense the first mixture into a solid dosage form. 1 8 · A pharmaceutical composition according to μ seeking item 7 wherein the composition is formulated to be stored at about &lt; 30 ° C and about &lt; 30% relative humidity for 24 months, including each bile 1 tocopherol The isomers are less than about 1% by weight. 19. The pharmaceutical composition according to claim 7, wherein the composition is formulated so that, after storage at about &lt; 30 ° C and about &lt; 30% relative humidity for 24 months, there are few degradation products including bile alcohol. At about 5% by weight. 20. A pharmaceutical composition comprising: a bisphosphate, a pharmaceutically acceptable salt, derivative, or hydrate of the bisphosphate, or a mixture thereof, cholestanol, wherein 'when administered for more than one week, the The effect of cholecalciferol is substantially similar to the effect of about 400 IU of cholecalciferol per day; moreover, the pharmaceutical composition is suitable for giving one dose per week. 2 1. The pharmaceutical composition according to claim 1, wherein the bisphosphate is sodium alendurate, and the blood concentration map of mammals derived from sodium alendurate administered with the composition is substantially similar to that obtained from administration of 70 Graph of plasma concentrations of milligrams of sodium alendurate in mammals without cholecalciferol. 22. The pharmaceutical composition according to claim 1, wherein the di-salt salt is sodium alenduric acid 92852.doc 200538136, and the plasma concentration map of mammals from cholecalciferol to which the composition is administered is substantially similar to that obtained from administration Graph of plasma concentrations of 2800 IU cholecalciferol in mammals without sodium hypothionate. 23. The pharmaceutical composition according to claim 1, wherein the serum concentration map of mammals for which the composition has been administered for at least one hour produces at least one of the following situations: Least squared (LS) mean AUC of cholecalciferol (at oi2Gh) It is about 296.4 nanograms per hour per milliliter, in which the serum concentration of the bile # 5 alcohol was not taken into account when measuring the pharmacokinetic parameters, and the average AUC (0_i20 hours) of the least square (LS) was about 297.5 nanograms · hour / ¾ liter, in which the pharmacokinetic parameters were measured using the concentration of cholecalciferol 0 hours before the dose as a covariate, and taking into account the baseline serum concentration of cholecalciferol, and ° Least squares (LS) average value AUQm, μ is about 143. 丨 Nanogram hours / ml, where the pharmacokinetic parameters are determined by subtracting the 120-hour estimated baseline langitol and taking into account Serum concentration of baseline linear alcohol. ° 24. According to the request! Pharmaceutical composition, in which the plasma concentration map of mammals for which the composition has been administered for ⑶ hours produces at least the following conditions:-Stable after 120 hours The maximum mean value of the maximum plasma inertia u in the steady state is about 5.9 nanograms per milliliter, of which one determines the pharmacokinetics. For several hours, the serum concentration of the baseline cholecalciferol was not considered. The maximum plasma concentration (c: the average value of the minimum squared 为 is about 5.9 nanograms / ml, so that when measuring the parameters, the serum cholestanol concentration 0 hours before the dose was used as: 92852.doc 200538136 variable, Taking into account the baseline serum concentration of cholecalciferol and the minimum square of the steady state maximum plasma concentration (cmax), it is about 4.0 nanograms / ml. Among them, the determination of the pharmacokinetic parameters minus the 120 hours after Estimated baseline cholecalciferol 'and the serum concentration of baseline cholecalciferol. "Consideration 25. A pharmaceutical composition according to request !, wherein the plasma indifference of mammalian bile pure alcohol after hours of administration" The maximum plasma Han degree of 120 is produced in Figure 120 (C ~ ~ ~ ~ ~ ~ ~ about 12 hours, in which the determination of the majority of the pharmacokinetics, did not take into account the baseline concentration of cholestyrol serum production. 26 .: = Item 1 Pharmaceutical composition, wherein In mammals, the composition 2 of the composition 3: the apparent median half-life of biliol alcohol plasma concentration is approximately estimated =, and, where the pharmacokinetic parameter is determined, the serum concentration subtracted is used.纟 Consider the baseline bile hearing 92852.doc