Tizanidine pyridinesulfonamide acid or their pharmaceutically acceptable salts or esters, pharmaceutical composition and method of treatment

 

(57) Abstract:

The invention relates to new tizamidine pyridinylmethyl acids f-ly R2-Z-Q-(CR1R1)n-CH[P(O)(OH)2]2(I) where R1-H, -SH, -(CH2)mSH or-S-C(O)-R3, R3- C1-C8-alkyl, m = 1 - 6, n = 0 to 6, Q is a covalent bond or-NH-, Z - pyridinyl, R2- H, -SH, -(CH2)mSH, -(CH2)mS-C(O)R3or-NH-C(O)-R4-SH, where R3and m have the above meaning, R4- C1-C8-alkylen, or their pharmaceutically acceptable salts or esters. The invention relates also to pharmaceutical compositions for treating or preventing diseases associated with abnormal calcium and phosphate metabolism, which contains a compound of the formula (I) in an effective amount and method of treatment of this disease. 3 S. and 7 C.p. f-crystals.

Background of the invention

The present invention relates to new nitrogen-containing, tizamidine heterocyclic phosphonate compounds, including bisphosphonates, phosphonocrotonate, phosphonocarboxylates and phosphosulfate.

This invention also relates to pharmaceutical compositions containing these is, characterized by abnormal calcium and phosphate metabolism, which uses the compound or pharmaceutical composition of the present invention.

Specifically, the present invention relates to a method of treating or preventing osteoporosis and arthritis, especially rheumatoid arthritis and osteoarthritis, when using the compounds or pharmaceutical compositions of the present invention.

A number of pathological conditions that affect warm-blooded animals, includes abnormal calcium and phosphate metabolism. Such States can be divided into two broad categories.

1. States that are characterized by abnormal mobility of calcium and phosphate, leading to General or specific lesion of bone, such as osteoporosis and Paget's disease, or to an exceptionally high content of calcium in the body fluids, such as hypercalcemia tumors.

Such States are sometimes referred to in this description of pathological demineralization of hard tissues.

2. Conditions that cause or are the result of abnormal deposits of calcium and phosphate in the body, such as rheumatoid arthritis and osteoarthritis. These States ibolee common metabolic bone disease osteoporosis osteoporosis is a condition in which solid bone collapse disproportionate to the formation of new hard tissue. In General, osteoporosis can be defined as the decline in the quality of bone or atrophy of skeletal tissue. And bone marrow cavity of bones become more reduced binding fibers, and a compact layer of bone becomes fragile.

Osteoporosis can be sub-classified as menopausal, senile caused by medicines (for example, adrenocorticoids that can be used when steroid therapy), caused by disease (arthritis and tumor), and so on; however, expression in both cases is almost the same. In General there are two types of osteoporosis: primary and secondary.

Secondary osteoporosis is a result of a separate set of painful process or agent. However, approximately 90% of all cases of osteoporosis is primary osteoporosis".

Such primary osteoporosis includes postmenopausal oreopoulos, (disuse) osteoporosis, age-related osteoporosis (affecting most individuals over the age of 70 - 80 years) and idiopathic osteoporosis, striking the m the rarefaction of bone tissue is high enough and leads to mechanical failure of the bone structure. Often bone fractures, such as hip and spine, in women suffering from postmenopausal osteoporosis. Kyphosis (hump) (abnormally increased curvature of the thoracic spine) can also be the result of osteoporosis.

The mechanism of bone destruction in osteoporosis is considered to include the deregulation process of bone rebuilding". Bone rebuilding occurs throughout life, it updates the skeleton and supports bone strength. This rebuild includes erosion and filling of the discrete areas on the surface of the bone with the help of an organized group of cells called the "basic multicellular units" or "OME". OME primarily consist of osteoclasts, osteoblasts and their cell precursors.

During the refresh cycle, the bone is resorbed in the area of "activated" OME with osteoclast with the formation of cavity absorption. This cavity is then filled with bone by osteoblasts.

Usually in adults update cycle leads to a slight deficiency of bone tissue vsledstvii zegenie bones. However, osteoporosis may increase the number of activated OME. This increased activation accelerates the renewal of bone tissue, leading to abnormally high bone loss.

Although the etiology of osteoporosis is still not fully clear, there are many risk factors that are associated with osteoporosis. These include low body weight, low calcium absorption, physical inactivity and estrogen deficiency.

Currently, the treatment of most cases of osteoporosis is calcium and estrogen.

The second category includes state, manifested in the abnormal deposition of calcium and phosphate, includes progressive mosinee bone formation (myositis ossificans progressiva), the total calcification, as well as diseases such as arthritis (including, for example, rheumatoid arthritis and osteoarthritis), neuritis, bursitis, tendonitis and other conditions that create a predisposition to the deposition of calcium.

In addition to osteoporosis, the bone loss may be a result of rheumatoid arthritis and osteoporosis. Rheumatoid arthritis is a chronic, systemic and sustavnoi inflammatory disease characterized by the weakening of the joint capsules and SWAT novelny liquid.

Symptoms of rheumatoid arthritis include General weakness, fatigue, localized pain, stiffness and weakness, and swelling and deformity of the joints of the body. Rheumatoid arthritis is the most common disease in women aged 40 to 60 years.

Pathogenesis of rheumatoid arthritis, leading to destruction of the joints, characterized by two phases:

1) exudative phase, including circulation and synovial cells, which allows flow into plasma proteins and cellular elements in the joint, and

2) chronic inflammatory phase flowing at subsynovial membrane, which is characterized by the formation of pannus (granulation tissue) in the joint and Podhradie bone, bone erosion and destruction of cartilage.

Pannus may lead to the merger and the scar tissue that causes deformity of the joint, is characteristic of rheumatoid arthritis.

The etiology of rheumatoid arthritis remains unclear. Infectious agents such as bacteria and viruses that play a significant role. Modern hypothesis is that the Epstein-Barr virus (EBV) is the agent causing rheumatoid arthritis.

Modern treatment reumatoideyanni drugs. Non-steroidal anti-inflammatory drug treatment is effective mainly in the early stages of rheumatoid arthritis; it is unlikely that it will suppress the inflammation of the joints, if the illness continues for more than one year.

Gold, methotrexate, immunosuppressants and corticosteroids were used with satisfactory results.

On the other hand, osteoarthritis is a congenital non-inflammatory disease of the synovial joints, which is characterized by attrition and abrasion of articular cartilage and formation of new bone on the surface of the joint. With the development of osteoarthritis, the surface of the articular cartilage is destroyed and worn particles gain access to the synovial fluid, which, in turn, stimulates phagocytosis by macrophage cells. Therefore, the inflammatory response ultimately occurs in osteoarthritis.

Common clinical symptoms of osteoarthritis include cartilage and bone growth of the finger joints and stiffness when waking up, and pain during movement. Conventional treatment of symptoms of osteoarthritis is the use of analgesics, anti-inflammatory agents, steroid-is in the treatment and prevention of diseases, including abnormal calcium and phosphate metabolism.

For example, a number of works that are included in this description as a reference, disclose compositions containing polyphosphonate, in particular, bisphosphonates, such as ethane-1-hydroxy-1,1-diphosphonic acid (AGDC), and their use for inhibiting anomalous deposition and mobility of calcium and phosphate in animal tissues: U.S. Patent 3683080, issued August 8, 1972, and U.S. Patent 4230700, issued October 28, 1980 (both Francis), U.S. Patent 4868164 (Ebetino), issued September 19, 1989

A number of other works describes a heterocycle-substituted diphosphonic acid, which can be used for the treatment of osteoporosis and/or arthritis: U.S. Patent 4868164 (Ebetino et al.), issued September 19, 1989; U.S. Patent 5104863 (Benedict et al.), issued April 14, 1992; U.S. Patent 4267108 (Blum et al.), issued may 12, 1981; European patent application (Boehringer Mannheim GmbH) N 170228, published February 5, 1986; European patent application N 186495 (Benedict and Perkins), published July 2, 1986; U.S. Patent 4754993 (Bosies et al. ), issued November 15, 1988; U.S. Patent 4939130 (Jaeggi et al.), issued July 3, 1990; U.S. Patent 4971958 (Bosies et al.), issued November 20, 1990; DE 4011777 (Jaeggi), published on October 18, 1990; application WO 90/12017 (Dunn et al.), published on October 18, 1990; application WO 91/10646 (Youssefyeh R., et al. ) Is And 4208401 (Bauman), issued June 17, 1980.

In addition, some studies have described sulfur-containing phosphonic acid, which, as indicated, can be used in the treatment of symptoms of inflammation (see, for example, U.S. Patent 4746654 (Breliere et al., transferred Sanoli), issued may 24, 1988; U.S. Patent 4876247 (Barbier et al.), issued October 24, 1989; and EPO 100718 (Breliere et al., transferred to Sanofi), published February 15, 1984.

Also in U.S. Patent 5071840 (Ebetino et al.), issued December 10, 1991, disclosed sulfur containing heterocycle-substituted diphosphonates in which diphosphonate-substituted carbon fragment is attached to the carbon atom in the nitrogen-containing six-membered heterocycle. Compounds described in the patent, can be used to treat conditions involving abnormal calcium and phosphate metabolism, especially osteoporosis and arthritis.

In addition, in the European patent 0298553 (Ebetino), published on 11 January 1989, describes tiltability along with a large number of other substituents, are acceptable as substitutes to methylentetrahydrofolate acids. However, do not indicate that tylny Deputy increases protivorazgonnoy and anti-arthritis activity in comparison with other described substituents.

Titlestyle described in this paper include thiol, alkylthiol, thioethers, alkylthiophene, dayevery and alkyldimethyl, THIOCARBAMATE, alkyldiethanolamine, dithiocarbamate, alkyldiethanolamine, thiocarbonate, alkylthiocarbamates, alkyldithiophosphate and dithiocarbonate.

In addition, the compounds of the present invention have osteotomies activity in the destruction of the joints in arthritis and have this activity as an additional positive effect in the treatment of arthritis as compared with the above simple weakening of the symptoms of inflammation.

The concept of "osteopata activity" used in this description, means of modifying disease activity on the bone and surrounding soft tissues of the site.

Unexpectedly found that compounds of the present invention are more potent bone protivorazgonnoy activity and greater therapeutic use in the treatment of osteoporosis and arthritis than heterocyclic bisphosphonate compounds that do not contain titlestatus. the developments and these compounds are potent inhibitors of bone resorption, useful in the treatment of osteoporosis, as well as anti-arthritis agents, useful in the treatment of osteoarthritis and rheumatoid arthritis.

An additional object of the present invention is to provide pharmaceutical compositions useful for the treatment and prevention of abnormal calcium and phosphate metabolism and for the treatment and prevention of arthritis, especially rheumatoid arthritis and osteoporosis.

In addition, an object of the present invention is to provide a method for treatment and prevention of diseases characterized by abnormal calcium and phosphate metabolism in humans and other mammals, including osteoporosis and arthritis, especially rheumatoid arthritis and osteoarthritis.

These and other objects of the present invention will become apparent from the detailed description of the invention, which is presented below.

The present invention relates to tizamidine nitrogen-containing heterocyclic phosphonate compounds, including bisphosphonates, phosphonocrotonate, phosphonocarboxylates and phosphosulfate, and their pharmaceutically acceptable salts and ester and an effective amount of the compounds of the present invention and pharmaceutically acceptable filler.

Finally, the present invention relates to a method of treatment or prevention of pathological conditions characterized by abnormal calcium and phosphate metabolism in humans or other mammals, including treatment or prevention of osteoporosis and arthritis, especially rheumatoid arthritis and osteoarthritis.

This method comprises the administration to a human or other mammal in need of such treatment, a safe and effective amount of the compounds or compositions of the present invention. These compounds have the following General formula:

< / BR>
where

R1- hydrogen group-SH, -(CH2)mSH or-S-C(O)-R3;

R3- C1-C8-alkyl;

m = 1 - 6, an integer;

n = 0 - 6, an integer;

Q is a covalent bond or a group-NH-,

Z - pyridinyl;

R2- hydrogen group-SH, -(CH2)mSH, - (CH2)mS-C(O)R3or-NH-C(O)-R4-SH, where R3and m have the above meanings;

R4- C1-C8-alkylen,

or their pharmaceutically acceptable salts or esters.

Acceptable thio-substituents in the compounds of the present invention are thiols, alkylthiol, thioethers, alkylthiophene, dayevery, alkelda, dithiocarbonate and alkalitolerant.

The present invention also relates to pharmaceutical compositions containing a safe and effective amount of a compound of the invention and a pharmaceutically acceptable filler.

Finally, the present invention relates to a method of treating or preventing pathological conditions characterized by abnormal calcium and phosphate metabolism in humans and other mammals. This method includes the introduction of the above for a person or other mammal in need of such treatment, a safe and effective amount of the compounds or compositions of the present invention.

Below is a list of definitions for terms used in this description.

"Alkyl" represents an unsubstituted or substituted, linear or branched, saturated or unsaturated hydrocarbon chain, and the above-mentioned hydrocarbon chain can be saturated, containing from 1 to 8 carbon atoms, and preferably, unless otherwise stated, from 1 to 4 carbon atoms; the above-mentioned hydrocarbon chain may be unsaturated, containing from 2 to 8 carbon atoms, and preferably, Elsa in this description, covers alkeneamine unsaturated hydrocarbon chain containing at least one olefinic double bond and alkyline unsaturated hydrocarbon chain containing at least one triple bond. Preferred alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl and butyl.

The term "bisphosphonate" or "bisphosphonic acid", which are also used in the present description relate to those phosphonates and phosphonic acids, which contain two phosphonate groups attached to the same carbon atom, and used interchangeably with the concepts of diphosphonate and diphosphonic acids.

"Pharmaceutically acceptable" salt is a cationic salt formed at any acidic group (e.g. carboxyl group), or an anionic salt formed at any basic group (e.g. amine).

In this area there are a large number of such salts, which are described in World patent application 87/05297 (Johnston et al.), published September 11, 1987, the Preferred cationic salts are salts of alkali metal (such as sodium and potassium) and alkaline earth salt of a metal (such as magnesium and calcium). Predpochtitaemye esters are tizamidine phosphate compounds, which do not interfere in the activity of compounds or which are easily metabolized in the body of man or other mammal with the release of active compounds.

A large number of such esters are described in World patent application 87/05297 (Johnston et al.), published on 11 September 1987 and included in this description as a reference.

Such esters are the lower alkalemia esters, lower aryloxyalkyl esters (such as acetoxymethyl, ecotoxicology, aminocarbonylmethyl, pivaloyloxymethyl and pivaloyloxymethyl esters), lactonase esters (such as Caligraphy and tittilicious esters), lower alkoxyalkanols esters (such as methoxycarbonylmethyl, ethoxycarbonylmethyl and isopropoxycarbonyloxymethyl esters), alkoxyalkyl esters, kalinovye esters, and acylaminoalkyl esters (such as acetamidomethyl ether).

As mentioned above, the substituting groups may themselves be substituted. Such substituents include, but are not limited to, the substituents listed in the work of C. Hansch, A. Leo, Substituent Constants for Correlation Analysis in Chemistry and Biology (1979), which is entered in the description as a reference, materilas, hydroxy, oxo, amino, aminoalkyl (for example, aminomethyl and others), cyano, halogen, carboxy, alkoxyalkyl (for example, carboethoxy and others), thio, thiol, aryl, cycloalkyl, heteroaryl, heteroseksualci (for example, piperidinyl, morpholinyl, pyrrolidinyl, piperazinil and the like.), imino, thioxo, hydroxyalkyl, aryloxy, arylalkyl, and combinations thereof.

Tizanidine nitrogen-containing heterocyclic phosphonate compounds.

Compounds of the present invention are tizamidine heterocyclic phosphonic acids and their pharmaceutically acceptable salts and esters in which the carbon atom containing the phosphonic acid residue attached to the carbon atom of nitrogen-containing heterocyclic fragment, preferably a pyridine ring.

The connection of the carbon atom containing the phosphonic acid residue, a heterocyclic residue may be carried out directly covalent bond, preferably a single bond) or with a chain length of from about 1 to 6 atoms.

If the connection is made through the chain, the chain may consist only of carbon atoms, represent a nitrogen atom or nitrogen-containing My carbon and nitrogen in binding chains can be independently from each other are not substituted or substituted by one or more substituents, choose from titlestyle (including thiols, alkylthiols, thioesters, alkylthiophenes, dityatev, acidification, thiocarbamates, architekturbuero, dithiocarbamates, alkyldiethanolamine, thiocarbonates, alkylthiophenes, dithiocarbonates and alkyldithiophosphate), hydrogen atom, hydroxy group, methyl, ethyl or propyl.

The carbon atoms and nitrogen atoms in the chain may be unsubstituted. Preferred chain with one atom, i.e.,- CH2-, -NH - and Acting

For compounds in which the nitrogen atoms, sulfur or oxygen in the linking chain attached to the heterocyclic fragment, this atom is nitrogen, sulfur or oxygen attached to the carbon atom of the cycle and not attached directly to the nitrogen atom of the cycle.

The present invention also includes those compounds in which the nitrogen atom in the linking chain is attached to the heterocyclic ring, where the nitrogen atom linked to the carbon atom directly adjacent to the nitrogen atom in the heterocycle, and these compounds have ridenow structure (which is more fully described below).

When Q represents NH, Q is preferably connected with the cycle of the carbon atom. is Yes, or to the nitrogen atom of the ring.

The carbon atom, which contains a phosphonate group may be unsubstituted (i.e., a hydrogen atom or substituted. The carbon atom may be replaced by two phosphonate groups (turning into bisphosphonate connection); or one phosphonate group and one phosphinate group (forming phosphonocrotonate connection); phosphonate group and a sulfate group (giving phosphosulfate connection); or phosphonate group and a carboxyl group (giving phosphonocarboxylate connection).

In addition, the carbon atoms in the heterocyclic ring can be unsubstituted or substituted independently of one another by one or more substituents. The nitrogen atom in the heterocyclic ring may also be substituted or unsubstituted.

Significant is the fact that the compounds of the present invention must contain at least one titlestyle, for example, -SH, or -(CH2)mSH. Accordingly, at least one of the substituents R1and R2should be-SH, or - (CH2)nSH.

In order to identify and assess pharmacological activity tests diphosphonate compounds n there.

Thus, bone antiresorptive activity in vivo can be demonstrated using a test designed to study the ability of these compounds to inhibit bone resorption and the bone resorption is characterized by abnormal calcium and phosphate metabolism.

Examples of such known test is the Schenk model in rats and adjuvant arthritic test. Also useful is the test in vitro for inhibition of crystal growth of hydroxyapatite. These and other suitable tests for the assessment of pharmacological activity are described and/or referenced in the works of Shinoda et al. , Calcified Tissue International, 35, pp. 87 - 99 (1983); Schenk et al., Calcified Tissue Research, 11, pp. 196 - 214 (1973); Russel et al. , Calcified Tissue Research, 6, pp. 183 - 196 (1970); Muhlbauer and Fleisch, Mineral Electrolyte Metab., 5, pp. 296 - 303; Nancollas et al., Oral Biol., 15, 731 (1970); U.S. Pat. USA 3683080 (Francis), issued on August 8, 1972; U.S. Pat. USA 4134969 (Schmidt-Dunker), issued January 16, 1979; and in the application of EPO 189662, published on August 6, 1986; all work included in this description as a reference.

Some of these tests for pharmacological activity are also described in more detail in the Examples below.

In addition to the use for the treatment or prevention of pathological conditions, characterposition and for other purposes. For example, the compounds of the present invention, it is believed, can be used as a bone scanning agent after labeling with 99m-technetium.

In addition, the compounds of the present invention can be used as an agent that binds ions of polyvalent metal chelate, preferably two ions (e.g., calcium ions and magnesium) and trivalent metals (e.g., India). Therefore, the compounds of the present invention can be used as components of detergents and cleaning agents, or for water treatment. They can also be used as stabilizers for compounds.

In addition, they can be useful in preventing the formation of Tartar (e.g., calculus) and/or Tartar. Finally, the compounds of the present invention can be used as herbicides that are toxic to animals.

Tizanidine nitrogen-containing heterocyclic phosphonate compounds of the present invention can be obtained according to the methods presented in Examples A-H.

Compositions containing the new tizamidine nitrogen-containing heterocyclic Vostok introduced to people or other mammals in different ways, including, but not limited to, using oral dosing forms or injection (intravenously, intramuscularly, intraperitoneally and subcutaneously).

A number of other dosing forms containing the new tizamidine compounds of the present invention, can be easily retseptoriani any qualified in this field specialist with the use of suitable pharmaceutical excipients, which are described below. From the point of view of convenience for the patient, the most preferred are oral dosing forms.

The term "pharmaceutical composition" as used in the description means a combination that includes a safe and effective amount tizamidine phosphonate compounds as the active ingredient, or mixtures thereof, and a pharmaceutically acceptable excipient.

The phrase "safe and effective amount" means such amount of the compound or composition, which is enough to significantly positively modify the symptoms and/or condition that must be treated, but small enough to avoid serious side effects (at a reasonable correlation of the positive EF is the active ingredient for use in pharmaceutical compositions which should be used in the method of the present invention will vary depending on the specific conditions being treated, the age and physical condition of the patient in need of treatment, severity of disease, duration of treatment, the nature of concurrent therapy, the specific active ingredient, the particular pharmaceutically acceptable carrier, and such factors are known and accounted attentive doctor.

The term "pharmaceutically acceptable carrier" as used herein, includes physiologically inert, pharmacologically inactive material known to the skilled in this field specialist, which is compatible with the physical and chemical characteristics of a specific phosphonate compounds selected as an active ingredient.

Pharmaceutically acceptable carriers are, but not limited to, polymers, resins, plasticizers, fillers, binders, lubricants, chemicals for slide, dezintegriruetsja substances, solvents, co-solvents, buffer systems, surfactants, preservatives, paudie viscosity.

The concept of "oral dosing form" means any pharmaceutical composition intended for the systematic introduction to the patient by delivery of the above composition in the gastrointestinal tract of the patient through the mouth.

For the purposes of the present invention form for delivery can be a tablet with shell or without shell, solution, suspension or capsule shell or without shell.

The term "injection" as used herein, refers to any composition intended for the systematic introduction to the human or other mammal by delivery of a solution or emulsion containing the active ingredient, when piercing the skin of the patient in order to introduce the above-mentioned solution or emulsion in the circulating system of the patient, by intravenous, intramuscular, intraperitoneal or subcutaneous injection.

The speed of the systematic introduction can be appropriately controlled qualified by manipulating one or more parameters from among the following:

(a) the actual active ingredient;

(b) pharmaceutically acceptable excipient, pascalc is also desired density and permeability (the ability to swell above excipient;

(d) time dependence of the state of excipient and/or within the excipients;

(e) the particle size of the granulated active ingredient;

(f) pH-dependent state of exipient.

In particular, solubility, acidity and sensitivity to hydrolysis of various tizanidine phosphonate active ingredients, such as acid additive salts, salts formed by the carboxyl group, for example, salts of alkali metals, salts of alkaline earth metals, etc. and ethers, for example, alkalemia, alkenilovyh, arrouye, Arakelova, can be used for guidance in selecting components.

In addition, acceptable pH-conditions must be created in the dosing form by adding buffer to the active ingredient in accordance with the desired model selection.

As mentioned above, pharmaceutically acceptable exipients (fillers) are, but not limited to, resins, fillers, binders, lubricants, solvents, substances to improve slip, dezintegriruetsja additives, co-solvents, surfactants, preservatives, sweetening agents, additives, corrective taste and smell, what="ptx2">

The preferred solvent is water.

Supplements, corrective taste and smell, which can be used in the present invention are additives described in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, 1990, pp. 1288-1300. Pharmaceutical compositions that can be used in the present invention contain 0 to 2% corrective agents.

Dyes or pigments that can be used are described in Handbook of Pharmaceutical Excipients, pp. 81 - 90, 1986, published by the American Pharmaceutical Association &the Pharmaceutical Society of Great Britain. The pharmaceutical compositions of the present invention contain 0 to 2% of the dye or pigment. Preferred co-solvents include, but are not limited to, ethanol, glycerin, propylene glycol, polyethylene glycol. The pharmaceutical compositions of the present invention contain 0 to 50% co-solvents. Preferred buffer systems include, but are not limited to, acetic, boric, carbonic, phosphoric, succinic, maleic, tartaric, citric, benzoic, lactic, glyceric, gluconic, glutaric, and glutamic acid and their sodium, potassium and ammonium salts. Especially preferred are phosphoric, tartaric, and 5% buffer systems.

Preferred surfactants include, but are not limited to, polyoxyethylenesorbitan esters of fatty acids, polyoxyethylenesorbitan esters, monetary sucrose and ethers and esters of lanolin, alkylsulfate salt, sodium, potassium and ammonium salts of fatty acids.

The pharmaceutical compositions of the present invention contain 0 to 2 percent surface-active substances.

Preferred preservatives include, but are not limited to, phenol, alkalemia esters by parahydroxybenzoic acid, o-phenylpropanamine acid and its salts, boric acid and its salts, sorbic acid and its salts, chlorbutanol, benzyl alcohol, thimerosal, acetate and nitrate of finalstate, nitromersol, benzalconi chloride, pyridinium chloride, methylparaben and propylparaben.

Especially preferred are salts of benzoic acid, pyridinium chloride, methylparaben and propylparaben. Compositions of the present invention typically contain 0 - 2% preservatives.

Preferred sweetening additives include, but are not limited to, sucrose, glucose, saccharin, sorbitol, mannitol and aspartame; especially predpochteniyami additives.

Preferred agents for increasing the viscosity include, but are not limited to, methylcellulose, sodium carboxymethyl cellulose, hypromellose, hydroxypropylcellulose, sodium alginate, carbomer, povidone, Arabian gum, diatomaceous earth, xanthan gum and tragakant.

Especially preferred are methyl cellulose, carbomer, xanthan gum, diatomaceous earth, povidone, sodium carboxymethylcellulose, and the silicate of magnesium. Compositions of the present invention contain 0 to 5% of an agent that increases the viscosity.

Preferred fillers include, but are not limited to, lactose, mannitol, sorbitol, rejonowy calcium phosphate, dibasic calcium phosphate, compressible sugar, starch, calcium sulfate, dextro - and microcrystalline cellulose. Compositions of the present invention contain 0 to 75% fillers.

Preferred lubricating agents include, but are not limited to, magnesium stearate, stearic acid, and talc. The pharmaceutical compositions of the present invention include a 0.5 - 2% lubricants.

Preferred substances for improving the slip are, but not oguche 1-5% substances to slip.

Preferred dezinfeciruyuhimi (dispersant) substances include, but are not limited to, starch, starch glycolate, sodium, crosspovidone, crosscarmellose sodium and microcrystalline cellulose. The pharmaceutical compositions of the present invention include 4 - 15% dispersant additives.

Preferred binders include, but are not limited to, Arabian gum, tragakant, hydroxypropylcellulose, pregelatinization starch, gelatin, povidone, hydroxypropylcellulose, hypromellose, methylcellulose, solutions of sugar, such as sucrose and sorbitol, as well as ethylcellulose. Compositions of the present invention contain 1 to 10% of binders.

Compounds of the present invention can comprise from about 0.1 to 99.9 wt.% the pharmaceutical compositions of the present invention. Preferably the compounds of the present invention comprise from about 15 to 95 wt.% compositions of the present invention.

Thus, the pharmaceutical compositions of the present invention contain 15 - 95% tizamidine phosphonate active ingredient or a mixture thereof, 0 to 2% corrective agents, 0 to 50% co-rasevic, 0 - 5% additives to improve viscosity, 0 to 75% filler, 0.5 to 2% connecting substances, 1-5% of the substances to improve slip, 4-15% dispersant additives, and 1 to 10% of binders.

The choice of pharmaceutical filler, which should be used in combination with tizamidine the phosphonates of the present invention, is fundamentally determined by the method of introduction of phosphonate compounds.

If the connection needs to be injected, the preferred pharmaceutical carrier is a sterile saline solution, pH of which is brought to approximately 7,4. However, the preferred method of introduction of the phosphonates of the present invention is oral and preferred dosing form is therefore tablets, capsules and other similar forms, containing from about 0.1 mg P to 600 mg P diphosphonic acids described herein.

Pharmaceutical carriers suitable for the preparation dosing unit forms for oral administration are well known in this field. Their choice will depend on the secondary consideration of taste, cost, stability, which is not critical to this invention and may beatie "mg P, used in this description, means the mass of phosphorus atoms, which are present in a number of diphosphonic acids of the present invention. This unit is used to standardize the number of diphosphonic acids of the present invention, which should be used in pharmaceutical compositions and method of the present invention.

For example, [[5-(2-mercapto-1-oxopropyl-2 - pyridinyl] aminomethyl] bis[phosphonic acid] has a molecular weight of 371 g/mol, where 16.7% (62 g/mol) due to the two phosphorus atoms present in the molecule. One milligram of this connection, therefore, contains 0.17 mg P (1 mg x 16.7%).

Thus, to prepare a pharmaceutical composition containing 1 mg P this compound, the composition should include 6 mg of the compound; and to provide a dose of 1 mg P/kg of this compound for a patient weighing 50 kg, the patient should take 300 mg of the compound.

Pharmaceutically acceptable carrier used in a mixture with phosphonates of the present invention, is applied in sufficient concentration to provide practical dosing. Preferably pharmaceutically acceptable carrier is from 20 to 80%.

Acceptable pharmaceutical compositions are presented in this description and in the Examples I - L. Qualified in this field specialist can produce changes in non-limiting examples, presented in the description, to obtain a wide range of pharmaceutical compositions.

A method of treating or preventing diseases characterized by abnormal calcium and phosphate metabolism.

Another object of the present invention is a method of treating or preventing diseases characterized by abnormal calcium and phosphate metabolism.

Such methods include administration to a human or lower animal in need of such treatment, a safe and effective amount diphosphonate compounds of the present invention.

The preferred method of administration is oral, but other known techniques can also be used, for example, namensliste (e.g., transdermal, rectal and parenteral (e.g. subcutaneous injection, intramuscular injection, intra-articular injection, intravenous injection, etc.). Can also be used inhalation.

Therefore, concrement, intravenous, intraperitoneal and subcutaneous methods, as well as local application method.

The concept of "abnormal calcium and phosphate metabolism" as used herein, means that [1] States that are characterized by abnormal mobility of calcium and phosphate, leading to General and specific cases of bone or excessively high content of calcium and phosphate in the body fluids; and [2] States that cause or are the result of abnormal deposits of calcium and phosphate in the body.

The first category includes, but is not limited to, osteoporosis, Paget's disease, hyperparasites, hypercalcemia malignant tumors, heterotropia ossification, and osteolytic bone metastases.

The second category includes, but is not limited to, progressive mosinee bone formation, total calcification, as well as diseases such as arthritis, rheumatoid arthritis, osteoarthritis, neuritis, bursitis, tendonitis and other conditions that create a predisposition for the deposition of calcium and phosphate in the tissues.

The term "rheumatoid arthritis" as used herein, refers to a chronic systemic and the United States, ligaments, tendons and bones.

The term "osteoarthritis" means a non-inflammatory disease of the synovial joints. It is characterized by attrition and abrasion of the articular cartilage and new bone formation on the surface of the joint.

The term "subject at risk" or "subject in need of such treatment" as used herein, refer to any human or other mammal, which has a significant probability of occurrence of abnormal calcium and phosphate metabolism, if left untreated, as well as to any person or other mammal, which established the presence of abnormal calcium and phosphate metabolism.

For example, women in the postmenopausal period, subjects exposed to a specific steroid therapy, subjects taking anticonvulsive drugs, subjects with Paget's disease, hyperparasitism, hypercalcemia malignant tumors or osteolytic bone metastases, subjects suffering from one or more of various forms of osteoporosis, the subjects belonging to groups whose risk of developing osteoporosis is significantly higher than average, for example, W is the n drugs which, as you know, as a side effect can cause osteoporosis, subjects suffering from progressive myositis the osteogenesis or General calcification, as well as subjects suffering from arthritis, osteoarthritis, neuritis, buritama, tendonitis and other inflammatory conditions that create a predisposition to the deposition of calcium and phosphate in the tissues.

The term "safe and effective amount" as used herein, means an amount of compound or composition of the present invention, high enough to significantly positively modify the condition to be treated, but low enough to eliminate the occurrence of side effects (at a reasonable correlation of the positive effect/risk) within the limits of medical law.

Safe and effective amount diphosphonate compounds of the present invention will vary depending on the specific condition that must be treated, the age and physical condition of the patient in need of treatment, severity of disease, duration of treatment, the Ki acceptable fillers, and similar factors within the knowledge and experience of the attentive physician.

However, single dosages can be in the range from about 0.01 mg P to 3500 mg P, or approximately from 0.0002 to 70 mg P/kg (based on body weight 50 kg).

Preferred single dosing the dose is a dose from about 1 mg P to 600 mg P or from about 0.02 to 12 g of P/kg of body weight (based on body weight 50 kg). Can be applied approximately to four single doses per day. Required daily dosage of about 500 mg P/kg in order to obtain the desired result, and a higher dose may cause unwanted effects.

Higher doses within the specified interval are required, of course, in the case of oral administration due to limited absorption.

The following additional Examples describe and illustrate a preferred embodiment within the scope of claims of the present invention. The examples are for illustrative purposes only and should not be construed as limiting the scope of the claims of the present invention, because of the possibly large number of changes in these examples, which do not affect the amount and existence is slots]

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The above connection get through the procedure described below.

I. Synthesis of tetraethyl ester [(5-bromo-2-pyridinyl) aminomethyl] bis[phosphonic acid].

2-Amino-5-bromopyridin (12.5 g, 72 mmol), triethylorthoformate (79.2 mmol) and diethylphosphate (158, 4 mmol) is heated at 140oC in a round bottom flask, equipped with a distillation attachment for collecting ethanol in the reaction. After heating for 8 h, the reaction mass is then cooled and then concentrated in vacuo. The desired product produce by rapid chromatography on silica gel using as eluent a 5% solution of isopropanol in methylene chloride.

II. Synthesis of tetraethyl ester [(5-(3-hydroxypropyl)-2 - pyridinyl)aminomethyl]bis[phosphonic acid].

To a solution of tetraethyl ester [(5-bromo-2-pyridinyl) aminomethyl] bis[phosphonic acid] (10 mmol) in THF (10 ml), cooled to - 78oC, add a solution of n-utility (2.1 equivalent) in hexane for 30 minutes, the Reaction mass is maintained at -78oC for a further 30 minutes

To the resulting solution was added C-itproportalmicrosoft ether (TMS) (2.5 equivalents) and the reaction mass allowed to warm up to comnatural-, TMS-ether)-2-pyridinyl)aminomethyl] bis[phosphonic acid] , which is used in the next stage without purification.

Removal of the TMS-ether of the molecules of the product is carried out by stirring it in THF with added dropwise within 30 min tetrabutylammonium fluoride (1M in THF). After standard aqueous processing allocate end of the primary alcohol in the form of oil and used in the next stage.

Ill. Synthesis of tetraethyl ester [(5-(3-bromopropyl)-2-pyridinyl)aminomethyl]bis[phosphonic acid].

A mixture of tetraethyl ester [(5-(3-hydroxypropyl)-2 - pyridinyl)aminomethyl] bis[phosphonic acid] (10 mmol), tetrabromide carbon (11 mmol) and triphenylphosphine (11 mmol) in dichloromethane (100 ml) was stirred at room temperature for 5 hours. Add water and the product extracted with dichloromethane. The combined organic extracts are dried and concentrated. The resulting residue is purified quick column chromatography, get tetraethyl ester [(5-(3-bromo-propyl)-2-pyridinyl)aminomethyl] bis[phosphonic acid].

IV. Synthesis of tetraethyl ester [(5-(3-acetylthiophene)-2-pyridinyl)aminomethyl]bis[phosphonic acid].

A solution of tetraethyl ester [(5-(3-br is by thioacetate sodium (5.2 mmol). The mixture was stirred at 50oC for 12 hours. After cooling to room temperature, evaporated in vacuo of the solvent. The crude residue is dissolved in methylene chloride and washed with water. The organic layer is then dried and concentrated in vacuo. The desired product was then purified by rapid chromatography on silica gel using as eluent 5 - 10% solution of isopropanol in methylene chloride.

V. Synthesis of [(5-(3-mercaptopropyl)-2-pyridinyl)aminomethyl]bis[phosphonic acid].

Thioacetate (4.2 mmol) is heated at the boiling point under reflux in 1N HCl (15 ml) for 5 hours. The reaction mixture is cooled, treated with charcoal, filtered and concentrated in vacuo. The target product of the desired degree of purity obtained when rubbing with acetone and with additional drying in vacuum over night.

Example b Synthesis of [(5-(3-acetylthiophene)-2-pyridinyl)aminomethyl] bis[phosphonic acid].

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[(5-(3-Acetylthiophene) -2-pyridinyl)aminomethyl] bis[phosphonic acid] obtained by heating at the boiling point under reflux tetraethyl ester [(5-(3-acetylthiophene)-2-pyridinyl) aminomethyl] bis[phosphonic acid] (received IU the camping was concentrated in vacuo and the product recrystallized from water and isopropanol.

Example C.

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The above compound is obtained according to the method described below.

I. Synthesis of tetraethyl ester [(5-nitro-2-pyridinyl) aminomethyl] bis[phosphonic acid].

2-Amino-5-nitropyridine (10 g, to 71.9 mmol), triethylorthoformate (11,7 g, 79,1 mmol) and diethylphosphate (21,86 g, 158,2 mmol) is heated at 140oC in a round bottom flask, equipped with a distillation attachment for collecting ethanol during the reaction. After heating for 10 h, the reaction mass is then cooled, and then concentrated in vacuo. The desired product is obtained with fast chromatography on silica gel using as eluent a 5% solution of isopropanol in methylene chloride.

II. Synthesis of tetraethyl ester [(5-amino-2-pyridinyl) aminomethyl]bis[phosphonic acid].

Tetraethyl ester [(5-nitro - 2-pyridinyl)aminomethyl]bis[phosphonic acid] (5.29 g, 12.4 mmol), absolute ethanol (100 ml) and 10% palladium on coal (1.3 g) was placed in a hydrogenation apparatus Parra 500 ml and hydronaut for 4 h at a pressure of 40 pounds per square inch.

The reaction mixture was filtered through celite, then concentrated in vacuo. The obtained solid product is used without further purification.

To nitrosodi tetrafluoroborate (NOBF4) (22 mg, 0,19 mmol) in methylene chloride (6 ml) at room temperature add tetraethyl ester [(5-amino-2-pyridinyl)aminomethyl]bis[phosphonic acid] (75 mg, 0,19 mmol).

The reaction mixture is stirred for 3 h and then concentrated in vacuo. The resulting crude residue is dissolved in acetonitrile (6 ml) and add sodium sulfide (46 mg, 0,19 mmol). After stirring for 12 h at room temperature the reaction is stopped by the addition of water and the mixture extracted with methylene chloride.

The organic extracts are combined and washed with 10% aqueous Na2S2O3. The organic extracts are then dried with sodium sulfate, filtered and concentrated in vacuo. The desired thiol get cleaning with fast chromatography, eluent of 2% isopropanol in methylene chloride.

IV. Synthesis of [(5-mercapto-2-pyridinyl)aminomethyl]bis[phosphonic acid] .

Bisphosphonic acid is obtained by boiling under reflux tetraethyl ester (0.5 mmol) in distilled water (25 ml) for 12 h under nitrogen atmosphere.

The reaction mass is treated with activated charcoal, filtered and concentrated in vacuo. Raw is th acid].

Example d Synthesis of [(4-(4-mercaptomethyl)-2-pyridinyl)aminomethyl] bis[phosphonic acid].

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The above connection get through the procedure described below.

I. Synthesis of tetraethyl ester [(4-bromo-2-pyridinyl) aminomethyl] bis[phosphonic acid].

Using the method of example A (part I) from 2-amino-4-bromopyridine, triethylorthoformate and diethylphosphate get tetraethyl ester [(4-bromo-2-pyridinyl) aminomethyl]bis[phosphonic acid].

II. Synthesis of tetraethyl ester [(4-(4-hydroxybutyl)-2 - pyridinyl)aminomethyl]bis[phosphonic acid].

To a solution of tetraethyl ester [(4-bromo-2-pyridinyl)aminomethyl] bis[phosphonic acid] (10 mmol) in THF (10 ml), cooled to -78oC, add a solution of n-utility (2.1 equivalent) in hexane over 30 minutes

The reaction mass is maintained at -78oC for a further 30 minutes To the resulting solution was added 4 - iodocholesterol ether (TMS) (2.5 equivalents) and the reaction mass allowed to warm to room temperature within 30 minutes After a standard water-treatment produce tetraethyl ester [(4-(4-butanol-, TMS-ether)-2-pyridinyl)aminomethyl]bis [phosphonic acid], which ISRI stirring it in THF with added dropwise within 30 min tetrabutylammonium fluoride (1M in THF). After standard aqueous processing allocate end of the primary alcohol in the form of oil and used in the next stage.

III. Synthesis of tetraethyl ester [(4-(4-acetyltributyl)-2-pyridinyl)aminomethyl]bis[phosphonic acid].

Using the same sequence of reactions described in Example A (part III-IV), tetraethyl ester [(4-(4-hydroxybutyl)-2-pyridinyl)aminomethyl] bis [phosphonic acid] in turn tetraethyl ester [(4-(4-acetylthio-butyl)-2-pyridinyl)aminomethyl]bis[phosphonic acid].

IV. Synthesis of [(4-(4-mercaptomethyl)-2-pyridinyl) aminomethyl] bis[phosphonic acid].

Thioacetate (5.0 mmol) is heated at the boiling point under reflux in 1N HCl (20 ml) for 8 hours. The reaction mass is then cooled, treated with charcoal, filtered and concentrated in vacuo. The desired product with the desired degree of purity receive subsequent rubbing with acetone and after further drying in vacuum.

Example e Synthesis of [(4-(4-acetyltributyl)-2-pyridinyl) aminomethyl] bis[phosphonic acid]

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[(4-(4-Acetyltributyl)-2-pyridinyl)aminomethyl]bis [phosphonic acid] obtained by heating at boiling point with obratnyye by the method of Example D, see above) in distilled will for 18 h in an argon atmosphere.

The reaction mixture was concentrated in vacuo and the product recrystallized from water and isopropanol.

Example f Synthesis of [[5-[(2-mercapto-1-oxopropyl)amino]-2-pyridinyl]aminomethyl] bis[phosphonic acid].

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The above connection receive in accordance with the methodology described below.

I. Synthesis of tetraethyl ester [[5-[(2-mercapto-1-oxopropyl) amino]-2-pyridinyl]aminomethyl]bis[phosphonic acid].

Timelocal acid (1,95 g, 18,38 mmol) is added slowly to the reagent pair hydrochloride 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (3,52 g, 18,38 mmol) in methylene chloride (15 ml) at 0oC. Then, to this mixture add tetraethyl ester [(5-amino-2-pyridinyl)aminomethyl] bis[phosphonic acid] [obtained by the method described in Example C (part II)] (4,84 g, 12,25 mmol) in methylene chloride (10 ml).

The reaction mixture was stirred at room temperature under nitrogen atmosphere for 24 hours. The reaction mass is diluted with methylene chloride (150 ml), then washed with water (2 x 150 ml), then saturated aqueous NaCl (1 x 125 ml).

The organic layer is dried sulfate NAT is anola in methylene chloride, and receives a yellow oil (3,05 g), yield 52%.

II. Synthesis of [[5-[(2-mercapto-1-oxopropyl)amino] -2-pyridinyl] aminomethyl]bis[phosphonic acid].

Tetraethylpyrophosphate (3,05 g of 6.31 mmol) is treated with bromotrimethylsilane (5,80 g, 37,89 mmol) in chloroform (25 ml) at room temperature under nitrogen atmosphere for 22 h.

The reaction is stopped by the addition of methanol and then the reaction mass was concentrated in vacuo. The crude residue is triturated with ethyl acetate and optionally dried in a high vacuum, get bisphosphonic acid (2,34 g) as a pale yellow solid, yield 100%.

Elemental analysis for C9H15N3O7P2S;

Calculated: C 29,11; H 4,07; N 11,32.

Found: C 28,48; H To 4.41; N 10,92.

Example g Synthesis of [2-acetylthio-2-(3-pyridinyl)ethylidene]bis[phosphonic acid]

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The above connection receive in accordance with the methodology described below.

I. Synthesis of 4,4'-(3-pyridinylmethyl)bomarton.

The suspension containing benzene (10 ml), 3-pyridine-carboxaldehyde (of 3.97 g, 37,09 mmol), boron trioxide (or 4.31 g, 61,94 mmol) and morpholine (7,76 g, 89,02 mmol), stirred at room temperature for 2 hours. The reaction mass is, what are square-73% (7,17 g) bisamides with a good degree of purity.

II. Synthesis of tetraethyl ester [3-(2-pyridinyl)attalides] bis[phosphonic acid].

To bicameral (1,00 g of 3.80 mmol) in toluene (6 ml) is added triperoxonane acid (0,89 g, 7,79 mmol). The mixture is heated for 15 min at 60oC add tetraethylethylenediamine (1.10 g, of 3.80 mmol) and the reaction mass is stirred for 22 h at 60oC.

The reaction mixture is cooled and water is added. The layers are separated and the aqueous layer was extracted with methylene chloride (3 x 15 ml). The organic layers are combined, dried with sodium sulfate, filtered and concentrated in vacuo. Bisphosphonate is separated from unreacted methylenediphosphonate and pyridinecarboxylic with fast chromatography on silica gel (eluent methylene chloride/isopropyl alcohol, 97:3).

Get the vinyl adduct (296 mg) with a yield of 20% in the form of a light yellow oil.

III. Synthesis of [3-(2-pyridinyl)attalides]bis[phosphonic acid]

Bisphosphonate (1.66 g, 4,39 mmol) is treated with bromotrimethylsilane (5,38 g, 35,12 mmol) in chloroform at 50oC for 12 h under nitrogen atmosphere. The reaction mixture was then stirred for 30 min with water (20 ml) and ethyl acetate (20 m bisphosphonic acid (0.66 g, yield 57%) as a pale yellow oil.

VI. Synthesis of [2-acetylthio-2-(3-pyridinyl)ethylidene]bis[phosphonic acid].

To [3-(2-pyridinyl)atenolole]bis[phosphonic acid] (0.56 g, 2,11 mmol) in water (5 ml) is added teoksessa acid (0,80 g, 10,55 mmol). After stirring at room temperature for 5 h, the reaction mass was concentrated in vacuo, triturated with acetone and then dried in high vacuum. Get bisphosphonic acid as a light yellow solid product (375 mg, yield 52%).

NAMR (D2O) 8,89 (1H, s); 8,71 (1H, m); 8,55 (1H, d, J=6 Hz); 7,88 (1H, m); the 5.51 (1H, dt, J=6,18 Hz); 2,85 (1H, dt, J=6,21 Hz); of 2.24 (3H, s).

PAMR (D2O) / 15,99; 15,29.

Example h Synthesis of [2-mercapto-2-(3-pyridinyl)ethylidene]bis[phosphonic acid].

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The above connection receive in accordance with the methodology described below.

I. Synthesis of tetraethyl ester [2-acetylthio-2-(3-pyridinyl) ethylidene] bis[phosphonic acid].

Tetraethyl ester [3-(2-pyridinyl)attalides] bis[phosphonic acid] (1.0 g, to 2.65 mmol) [prepared according to the procedure described in Example G (part II)] and teoksessa acid (0,30 g, 3,98 mmol) stirred in anhydrous chloroform (15 ml) for 48 h at room temperatute the second time in a vacuum, get thioacetate (1.01 g, yield 83%).

II. Synthesis of [2-mercapto-2-(3-pyridinyl)ethylidene]bis[phosphonic acid].

Bisphosphonic acid is obtained by heating under reflux tetraethyl ester [2-acetylthio-2-(3-pyridinyl) ethylidene]bis[phosphonic acid] (1.01 g, 2.21 mmol) in concentrated hydrochloric acid for 3 hours.

The solution is then evaporated to dryness in a vacuum. The crude residue is dissolved in warm water and treated with charcoal, then filtered through celite. The aqueous filtrate is extracted twice with methylene chloride. The product precipitated from the aqueous filtrate by adding ethanol. The precipitate is filtered off, washed with diethyl ether and dried in vacuum desiccator.

Elemental analysis for C7H11NO6P2S:

Calculated: C 28,46; H 3,90; N 4,53.

Found: C 28,44; H 3,93; N To 4.52.

Example I. Synthesis of [5-mercapto-2-(3-pyridinyl) pentylidene]bis[phosphonic acid]

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The above connection receive in accordance with the methodology described below.

I. Synthesis of ethyl ether, tert-butyldimethylsilyl ester 5 - hydroxy-2-(3-pyridinyl)pentanol acid.

To a solution of ethyl 3-pyridylacetate (0,76 g, 4,60 mmol) in a Solution is stirred for 30 min at -78oC and then to this solution was added 3-iodopropane, tert-butyldimethylsilyloxy ether (5.00 mol) in THF (20 ml).

The reaction mass was stirred at -78oC additional 2 hours and then at room temperature for 8 hours. The reaction is stopped by adding an aqueous solution of ammonium chloride. The layers are separated and the aqueous layer was extracted with diethyl ether. The organic layers are combined, dried and concentrated. The product was then purified by rapid chromatography on silica gel (eluent 20% methylene chloride in hexane).

II. Synthesis of 5-tert-butyldimethylsilyl ether 2-(3-pyridinyl) pentane-1,5-diol.

Carboxylate (2.25 mmol) reduced to the corresponding alcohol by treatment with lithium aluminum hydride (5,50 mmol) by boiling under reflux in THF (100 ml) under nitrogen atmosphere. The reaction is stopped by careful addition of water followed by treatment of the aluminum salts diluted aqueous solution of NaOH.

The reaction mass is filtered through celite and then the layers separated. The aqueous layer was extracted with diethyl ether. The organic layers are combined, dried and concentrated in vacuo. The oil obtained is used without further purification.

III. Synthesis of tert-butyldimethylsilyl who yl)pentane-1,5-diol (10 mmol), tetrabromide carbon (11 mmol and triphenylphosphine (11 mmol) in dichloromethane (100 ml) was stirred at room temperature for 5 hours. Add water and the product extracted with dichloromethane. The combined organic extracts are dried and concentrated. The residue is purified by rapid chromatography on a column and get a tert-butyldimethylsilyloxy ester 5-bromo-4-(3-pyridinyl)pentanol.

IV. Synthesis of diethyl ether, tert-butyldimethylsilyl ester 5-hydroxy-2-(3-pyridinyl)Interfactional acid.

A solution of tert-butyldimethylsilyl ester 5-bromo-4-(3 - pyridinyl)pentanol (0.75 mmol) and triethylphosphite (1.12 mmol) maintained at 90oC for 72 h, keeping the reaction mixture in a stream of nitrogen. Excess trimethylphosphite distilled off and the crude residue chromatographic on silica gel using as eluent 2%- aqueous solution of isopropanol in methylene chloride. The resulting product is used in subsequent reactions without further purification.

V. Synthesis of diethyl ether, tert-butyldimethylsilyl ester [5-hydroxy-2-(3-pyridinyl) pentylidene]bis[phosphonic acid].

To a solution of diethyl ether, tert-butyldimethylsilyl ester 5-hydroxy-2-(3-pyridinyl)pencilfake) at 0oC. After the addition stirring is continued for another 30 minutes the resulting solution was slowly added to a solution of diethylphosphate (2.50 g, 14,47 mmol) in anhydrous THF (100 ml) at room temperature.

After stirring the reaction mixture overnight, the reaction stopped by the addition of saturated aqueous sodium bicarbonate solution and the reaction mass is extracted with methylene chloride. The combined organic extracts are dried with anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product is purified by rapid chromatography on silica gel using as eluent 30% solution of acetone in hexane.

VI. Synthesis of diethyl ester [5-hydroxy-2-(3-pyridinyl) pentylidene] bis[phosphonic acid]

Silloway ether (0.50 mmol) were cleaved by treatment with tetrabutylammonium fluoride (0.75 mmol) in THF at room temperature for 30 minutes After removal of the protection reaction mass is washed with saturated NaCl solution. The organic layer is dried with sodium sulfate, filtered and then concentrated in vacuo. The resulting residue is used without further purification.

VII. Synthesis of diethyl ester of [5-bromo-2-(3-pyridinyl) pentylidene] bis[phosphono] bis[phosphonic acid] turn in diethyl ether [5-bromo-2-(3-pyridinyl)pentylidene]bis[phosphonic acid].

VIII. Synthesis of diethyl ester of [5-acetylthio-2-(3 - pyridinyl)pentylidene] bis[phosphonic acid].

A solution of diethyl ester of [5-bromo-2-(3-pyridinyl)pentylidene] bis[phosphonic acid] (5.0 mmol) is stirred in dry acetone (35 ml) and add thioacetate sodium (5.2 mmol). The mixture was stirred at 50oC for 12 hours. After cooling to room temperature the solvent is evaporated in vacuo, the resulting crude residue is dissolved in methylene chloride and washed with water.

The organic layer is then dried and concentrated in vacuo. The desired product was then purified by rapid chromatography on silica gel using as eluent a gradient of 5 to 10% solution of isopropanol in methylene chloride.

IX. Synthesis of [5-mercapto-2-(3-pyridinyl)pentylidene]bis[phosphonic acid] .

Diethyl ester of [5-acetylthio-2-(3-pyridinyl)pentylidene] bis[phosphonic acid] (4.2 mmol) is dissolved in 2.5 M hydrochloric acid (65 ml) and heated at boiling temperature under reflux for 7 hours. The reaction mass is then cooled and concentrated in vacuo. The solid residue triturated in acetone and then crystallized from ethanol / water. Get [5-mercapto-2- (3-pyridinyl)pentylidene]bis[phosphonic acid]and inhibition of bone resorption and inhibition of mineralization by using a model system in animals, known in the field of bone metabolism as a model of Schenk.

The General principles of this model system are revealed in the works of Shinoda et al., Calcif. Tissue Int., 35, 87 - 99 (1983); - and Schenk et al., Calcif. Tissue Res. II, 196-214 (1973), which are introduced in this description as a reference.

Materials and methods.

Animals.

Feeding on mother's milk 17-day (30 g) of male rats Sprague Dawley (Charles River Breeding Laboratories) are transported together with females and after arriving placed in a plastic chamber with females. At the age of 19 days of the web, follow, receiving rat food and water adlibitum randomly assigned to the subjects and the control group, including seven animals per group.

On the first day and on the 7th day again all animals injected intraperitoneally (b) Calcein (1% solution in 0.9% saline solution, the dose of 0.2 ml/100 g body weight). On day 4, all animals inject hydrochloride tetracycline (1% solution in 0.9% saline solution, the dose of 0.2 ml/100 g body weight).

These compounds serve as markers actively mineralizuyushchie (caliciviruses) bone and cartilage.

Dosage solutions and methods of dispensing.

All solutions prepared for subcutaneous injection in 0.9% northfleet based on consideration of the mass of powder (based on molecular weight, hydration) of the active material in mg/kg (body weight), which corresponds mg P/kg Concentrations are based on doses of 0.2 ml/100 g body weight. Usually all connections are assigned at the rate of 0.01, 0.1, 1.0, and 10.0 mg P/kg/day for 7 days.

The compounds exhibiting activity at 0.1 mg P/kg/day, then feel logarithmic reduction of the dose of 0.001 mg P/kg/day. The regulation of the dose, taking into account changes in body weight are conducted through the daily dosage.

The autopsy, tissue and histomorphometry.

On the 8th day after the start of injection of the compounds all animals wordplays by injecting intraperitoneally of overdose pentobarbital. The tibia is cut and placed in 70% ethyl alcohol. One large tibia dehydration in solutions of ethanol and contribute to the methyl methacrylate, as described by Schenk in the work Methods of Calcified Tissue Preparation (G. R, Dickson, Editor; Elsevier Science Publ., The Netherlands, 1984), which is introduced in this description as a reference.

Tibia-bone cut longitudinally through metafisico zone. According to one of the surfaces of the samples stained with silver nitrate and placed onto a glass slide for evaluation using a Quantimet Image Analyzer (Cambridge Instruments, Inc.), with COI is hydrated composition is measured in the area between the fluorescent label and plate neoplasms: is expressed as a percentage of total area (bone + bone marrow). The width of the epiphyseal plate neoplasms obtained as the average value of 10 equally oriented measurement cross-section.

Statistical evaluation of the data makes using parametric and non-parametric analysis of variations and test Wilcoxons rank sum for the evaluation of a statistically significant effect compared to control animals.

Using the model Schenk get the data for the in vivo inhibition of bone resorption by using compounds of the invention. The lowest effective (antiresorptive) dose (NED) for concrete tested compounds, which are determined by the model of Schenk.

Example K. Adjuvant arthritic model.

There are a number of examples of modeling of arthritis in animals, for example, provoked by the adjuvant arthritis, which is Mycobacterium butyricum.

This model represents one of the ways mimic rheumatoid arthritis in humans (swelling of the joints associated with the inversion of cells and synovial pannus plane, bone resorption and release of hemotoxicity factors and liposomal components in sustavnoi space) [1, 2].

A number of preventive and therapeutic research is nutov [5, 6].

Links

1. Pearson, S. , Wood F. (1959), Studies of Polyarthritis and Other Lesions Induced by Injection of Mycobacterial Adjuvant. 1. General Clinical and Pathological Characteristics and Some Modifying Factors, Arth. Rheum., 2:440-459.

2. Blackman, A., Burns, J. W.,*Radziwonik H., Westwick J. (1977), An X-ray Analysis of Adjuvant Arthritis in the Rat. The Effect of Prednisolone and Indomethacin, Agents and Actions, 7:145-151.*Framer J. B.

3. Winter C. A., Nuss, G. W. (1966), Treatment of Adjuvant Arthritis in the Rat with Anti-inflammatory Drugs, Arth. Rheum., 9:394-404.

4. Winder, S. V., L. Lembke, A., Stephens, M. D. (1969), Comparative Bioassay of Drugs in Adjuvant-Induced Arthritis in Rats: Flufenamic Acid, Mefenamic Acid, and Phenylbytazone, Arth. Rheum., 12:472-482.

5. Francis M. D., Flora L., King, W. R. (1972), The Effects of Disodium Ethane-l-Hydroxy-l-Diphosphonate on Adjuvant Induced Arthritis in Rats, Calcif. Tiss. Res., 9:number 109-121.

6. Flora L. (1979), Comparative Antiinflammatory and Bone Protective Effects of Two Diphosphonates in Adjuvant Arthritis, Arth. Rheum., 22:340-346.

Adjuvant arthritis is a severe cellulite and synovitis induced in male rats (or line Sprague Dawley or line Lewis) with a single subcutaneous injection (p/C) Mycobacterium butyricum (8 mg/ml) in mineral oil at 0 day.

Compounds are dosed once daily or oral (p/o), or parenteral (p/C) and can be tested for prophylactic (0 days) or therapeutic (9 - 10 or 14 days) modes.

Antartica efficiency can be measured as a reduction of the volume of the paws, bootanim saline solution. Processing can be stopped with further study of the "sudden flare-up" (the rapid growth of inflammation), which assess the ability of a joint to keep active.

Materials and methods.

A. Animals.

Used male Lewis rats (LEW). Upon arrival, rats randomly distribute the received data on a computer random number and put in a separate suspended on a wire chambers. Food and water are introduced ad libitum throughout the study. The care and safety of animals provide in accordance with State and Federal law. Each rat is identified by the number that is put in front of the camera and on the tail of a rat.

B. Experimental methodology.

First of laziness for all animals measured body weight (W) and the volume of the hind paws [(OL), recorded by the method of displacement of mercury using a pressure sensor connected to the computer].

At day 0 in the following way cause arthritis using MFA (Mycobacterium butyricum (Mb) 4.4 mg/kg in oil): rats anaesthetize and hold unit s/C injection MFA at the base of the tail under aseptic conditions.

The volume of the paws and the weight of the solids is measured in different days, about the with and the processing beginning at day 0 and continued daily until the end of the experiment. When the mode of treatment of rats randomly distributed into groups of 8 to 10 rats in accordance OL on day 10.

Dosing starting on day 10 and continuing daily until the end of the experiment. When both modes on day 10 or after 10 days, animals are placed in Shoe boxes with deeply located bedding.

Dosing solutions for connections that cannot be oxidized.

The drugs weighed on the calibration scale and then mixed in a volumetric flask with distilled water. The solution is brought to a pH of 7.4 using O 1N NaOH. Then the solution is filtered through a 0.45 μm sterile filter into a sterile container for storage. If the solution is not in use, store in the refrigerator.

For compounds that can be oxidized.

The drugs weighed on the calibration scale and then mixed in a volumetric flask with deionized water. The original solution is filtered through a 0.45 μm sterile filter into a sterile container for storage. If the solution is not in use, store in the refrigerator.

Based on the daily demand a specific amount of solution is withdrawn from the base solution, placed in a small measuring a beaker and then bring Velich the e dilution of the solution set pH (using deionized water).

Calculation of medicines is carried out on the basis of molecular weight, degree of purity of the compounds, the number is based on mg/kg (body weight) and the desired final concentration in mg P/kg Volume per rat is 0.1 ml/100 g body weight subcutaneously obtained in the form of injection in the groin crease of the animal in different directions every day or 1 ml/200 g of W obtained orally using curved dosing cutting stainless steel.

Weekly, based on the changes of body weight, spend an adjustment of the doses.

Chest x-ray, autopsy and collection of tissues.

At the end of the experience every rat killed by administering intraperitoneally with 1 ml Socomb. Immediately get x-rays whole body using x-ray element Torrox at 1200 MA = 5, ISUP = 50 and at time = 60 sec on medical unshielded film Kodak.

The hind limb of each rat was separated and fixed in 10% buffered formalin together with pieces of liver, kidney, spleen and thimus. Tibiotarsal joints decalcification using 4% EDTA at pH 7.4 and transferred in the usual way in paraffin blocks and H + E stain.

Histological part of the evaluated quantitatively for payout on bone resorption (CR) 6 anatomical trabecular bone parts in each hind foot and 4 sections on each front paw on a scale 0 - 3, giving the conditional score 0 - 60 for all four limbs.

To reactivate the formation of new bone (RNA) radiograph graduate on a scale of 0 - 3 for secondary and medical surface of the tibia, and then on a scale of 0 - 2 for all other surfaces, which are mentioned above, assigning conditional score 0 - 44.

D. Statistical analysis.

Data analysis the volume of the paw, bone resorption and reactivated the formation of new bone is carried out using student's t-test and single-path analysis of variations with Tukeys (SAS) [12]. Differences are considered significant at a pH of 0.05 or below.

This model provides in vivo data on the effectiveness of antiarthritic compounds from the viewpoint of reducing the swelling of the bones of the legs and reactivated new bone formation compared to arthritic animals treated with saline.

Example L. Capsules prepared by conventional methods, and they contain the following components:

Active ingredient - mg capsule

[5-Mercapto-2-(3-pyridinyl)pentylidene]bis[phosphonic acid] - 350,0

Fillers

Lactose - 99,0

Microcrystalline cellulose - 60,0

Magnesium stearate - 1 is Anna below:

The active ingredient is mixed with microcrystalline cellulose in a rotating drum mixer for approximately 10 minutes

The resulting mixture is passed through a hammer mill with a screen of 80 mesh.

Mix together with lactose again placed in the mixer with dual drum and stirred for approximately 15 minutes

Next, add magnesium stearate and mix for another 5 minutes the resulting mixture is then compressed in a reciprocating device for filling capsules.

Example M. to Prepare tablets having the following composition:

The active ingredient mg per tablet

[5-Mercapto-2-(3-pyridinyl)ethylidene]bis[phosphonic acid] - 700

Fillers

Lactose - 200

Starch (1500) - 100

Magnesium stearate - 25

Tablets having the above composition is prepared in the usual way, which is described below:

The active ingredient is pulverized in a ball mill for about 30 minutes Powdered active ingredient is then mixed for about 20 min in a two-blade mixer with lactose, dried by spraying.

To the resulting mixture add the starch and the mixture is additionally stirred for 15 minutes the Mixture preemie 6 months, significantly reduce bone resorption in a patient weighing approximately 70 kg, suffering from Paget's disease.

Similar results were obtained when replacing in the above-described tablets [2-mercapto - 2-(3-pyridinyl)ethylidene] bis[phosphonic acid] to [(5-(3 - mercaptopropyl)-2-pyridinyl)aminomethyl] bis[phosphonic acid]; [(5-(3-acetylthiophene)-2-pyridinyl)aminomethyl]bis[phosphonic acid]; [(5-mercapto-2-pyridinyl)aminomethyl] bis[phosphonic acid] ; [(4-(4-acetyltributyl)-2-pyridinyl)aminomethyl] bis[phosphonic acid]; [(4-(4-mercaptomethyl)-2-pyridinyl)aminomethyl] bis[phosphonic acid]; and their pharmaceutically acceptable salts or esters.

Example N. the Solutions for injection are prepared by conventional means using 10.0 ml of physiological solution and 7.0 mg of P - [2-mercapto-2-(3-pyridinyl)ethylidene]bis[phosphonic acid], bringing the pH to 7.4.

One injection once a day for 4 days results in a measurable relief of hypercalcemia malignant tumors in patients with a body weight of approximately 70 kg

Example o Male-Caucasian weighing approximately 92 kg 72 years of age suffer from moderate to severe pain, as well as from periodic swelling pret doctor which for the clinical diagnosis sets osteoarthritis of the right knee, which was later confirmed by x-ray study.

After a period of improving the treatment of various NSAID-AMI, including aspirin, naproxenum and Ketoprofen, the symptoms continue to worsen and worsen. He returned to his doctor, who prescribes him the tablets prepared in accordance with example M, for the reception twice a day for two hours before or two hours after meals for three months.

Clinical symptoms of pain and swelling of the joints, especially during prolonged walking, significantly improved after three months of treatment. After three months of treatment continued indefinitely at a dose of 1 capsule prepared according to the method of example L, laziness, at half the dose prescribed initially (that is 1 capsule in laziness).

Example P. Female black race weighing about 65 kg 55 years of age suffer from swelling and deformity of the joints of the fingers of both hands with partial loss of strength and/or flexibility in their fingers and hands. After visual and x-ray examinations, as well as after various clinical trials conducted P> After unsuccessful treatment with analgesics and anti-inflammatory agents, the doctor prescribes her tablets prepared according to Example M, twice a day for two hours before or after a meal for four months. After treatment in the course of a month, the symptoms associated with swelling of the finger joints, improved noticeably and significantly increases the mobility of the fingers; she continues treatment for the remaining four months, after which her doctor prescribes continued treatment prescribed doses for 2 months.

Example Q. a Girl of Spanish origin 12 years of age, weighing approximately 37 kg, go to the doctor with idiopathic juvenile rheumatoid arthritis. Symptoms include inflammation of the large number of joints, accompanied by fever and pain, which indicates a quick and pathological degradation of function of joints.

The doctor sends her to a rheumatologist, who immediately prescribes her intensive treatment by introducing the mode IV solution prepared according to Example N, for three days at the dose of 1 injection per day, administered within 2 hours.

At the end of the fourth mode, the physician prescribes taking two tablets, p is of which there is a significant improvement, accompanied by the increase in mobility and decrease pain.

After two months the doctor reduces the dose to 3/4 of the original oral dose of 3 tablets prescribing within two days, once 2 capsules a day, and once 1 capsule per day.

At the end of this mode, the dose again reduced to 1/4 of the initial dose, giving the patient a capsule prepared according to Example L, based on 1 capsule each day for an additional four months.

1. Tizanidine pyridinesulfonamide acid of the General structural formula

< / BR>
where R1- hydrogen group-SH, - (CH2)mSH or-S-C(O)-R3;

R3- C1-C8-alkyl;

m = 1 - 6, an integer;

n = 0 - 6, an integer;

Q is a covalent bond or a group-NH-;

Z - pyridinyl;

R2- hydrogen group - SH, -(CH2)mSH, -(CH2)mS-C(O)-R3or-NH-C(O)-R4-SH, where R3and m have the above meanings;

R4- C1-C8-alkylen,

or their pharmaceutically acceptable salts or esters.

2. Connection on p. 1, where R1represents a mercapto group.

3. Connection on p. 1, where R2represents the atom bogorodceani, associated with abnormal calcium and phosphate metabolism in humans or other mammals, containing the active ingredient and pharmaceutically acceptable excipient, characterized in that as the active ingredient it contains an effective amount of compounds of formula I.

5. The composition according to p. 4, wherein the disease associated with abnormal calcium and phosphate metabolism in humans or other mammals, is arthritis.

6. A method of treating or preventing diseases associated with abnormal calcium and phosphate metabolism, characterized in that designate a person or other mammal in need of such treatment, a safe and effective amount of the compounds under item 1.

7. The method according to p. 6, characterized in that a person or other mammal suffering from osteoporosis.

8. The method according to p. 6, characterized in that a person or other mammal suffering from arthritis.

9. The method according to p. 6, characterized in that a person or other mammal suffering from osteoarthritis.

10. The method according to p. 9, wherein the human or other mammal suffering from rheumatoid arthritis.

 

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< / BR>
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