HUT69732A - Quaternary nitrogen-containing phosphonate compounds, for treating abnormal calcium and phosphate metabolism - Google Patents

Description

The present invention relates to novel quaternary nitrogen-containing phosphonate compounds, including diphosphonates, phosphonoalkylphosphinates, phosphonocarboxylates and phosphonosulfates. The invention also relates to pharmaceutical compositions containing the novel compounds, and to a method of treating or preventing certain metabolic-related bone disorders characterized by abnormal calcium and phosphate metabolism using a compound or pharmaceutical composition of the invention. Specifically, the invention relates to a method of treating or preventing osteoporosis and arthritis, in particular rheumatoid arthritis and osteoarthritis / osteoarthritis, by use of a compound or pharmaceutical composition of the invention.

Many of the pathological conditions that can torture warm-blooded animals are associated with abnormal calcium and phosphate metabolism. These conditions fall into two broad categories:

1. conditions characterized by abnormal calcium and phosphate motility leading to general or specific bone loss, such as osteoporosis and Paget's disease; or excessively high levels of calcium and phosphate in body fluids, such as tumor-derived hypercalcemia. These are sometimes referred to herein as demineralization / desalting of abnormal calcified tissues;

> - 3 *

2. conditions caused by or as a result of abnormal deposition of calcium and phosphate in the body such as arthritis / arthritis / predominantly rheumatoid arthritis and osteoarthritis / osteoarthritis /. These conditions are sometimes referred to as abnormalities.

The first category includes the most common osteoporosis, osteoporosis. Osteoporosis is a condition in which the loss of calcified bone tissue is disproportionate to the formation of new calcified / hardened / tissue. Osteoporosis can generally be defined as a decrease in bone volume or skeletal tissue atrophy. The spaces between the bone marrow and the bone become larger, the fibrous bond is reduced, and the solid bone is fragile. Osteoporosis can be caused by cessation of menstruation, old age, medication (e.g., adrenocorticoid, which may occur with steroid therapy), disease / arthritis and tumor / etc, but the manifestation of the disease is essentially the same.

There are generally two types of osteoporosis: primary and secondary osteoporosis. Secondary osteoporosis is the result of another disease or agent. However, about 90% of all osteoporosis cases are primary osteoporosis. Primary osteoporosis includes post-menstrual osteoporosis, age-related osteoporosis, osteoporosis due to non-use (this affects most 70-80 year olds), and «

* ·· • · · ··· • ·

- 4 primary / spontaneous / osteoporotic disorders affecting middle-aged and younger men and women.

In some individuals with osteoporosis, bone loss is large enough to cause mechanical failure in bone structure. Bone fractures are common, for example, in women with postmenopausal osteoporosis in the hip and spine. Pupus may also occur (abnormally increased curvature of the thoracic spine).

The mechanism of bone loss in osteoporosis most probably involves an imbalance in the process of bone remodeling. Bone renewal takes place throughout life, renewing the skeleton and maintaining the strength of the bones. This regeneration involves the erosion and filling of individual sites on the bone surface by an organized / organized / group of cells, the basic multicellular units / or BMUs. BMUs consist primarily of osteoclasts, osteoblasts and their cellular precursors. In the cycle of remodeling, bone is absorbed at an activated site of the BMU by an osteoclast / bone disrupting cell, forming a resorption cavity. This cavity is then filled with bone by an osteoblast.

The regeneration cycle in adults usually results in a small loss of bone due to imperfect filling of the resorption cavity. Therefore, even in healthy adults, age-related bone loss occurs. Osteoporosis

however, individuals may experience an increase in the number of activated BMUs. Increased activation accelerates bone remodeling, resulting in abnormally high bone loss.

Although the cause of the disease is not fully understood, there are many risk factors that are probably related to osteoporosis. These include low body weight, low calcium intake, physical inactivity and estrogen deficiency.

Current treatment for osteoporosis consists primarily of calcium and estrogen.

The second category, caused by an abnormal deposition of calcium and phosphate, manifests itself in several diseases, such as myositis ossificans progressive myositis, calcinosis universalis, and diseases such as arthritis and rheumatoid arthritis osteoarthritis (osteoarthritis), neuritis (neuritis), bursitis (mucosal inflammation), tendonitis (inflammation of the mucosa), and conditions that tend to cause the tissue in question to deposit calcium.

In addition to osteoporosis, bone loss can also result from rheumatoid arthritis and osteoarthritis. Rheumatoid arthritis is a chronic, systemic, and arthritic disorder characterized by weakening of the articular capsules and ligaments, followed by destruction of cartilage, ligaments, in and bone, and other changes in the viscosity of the joint fluid. Symptoms of rheumatoid arthritis include general weakness, fatigue, stationary pain, and stiffness, weakness, swelling and deformation of the joints of the body. Rheumatoid arthritis is very common in women between the fourth and sixth decades of life.

The pathogenesis of rheumatoid arthritis leading to joint destruction is characterized by two phases: 1 / an exudative / swollen / phase, which involves microcirculation and synovial / synovial fluid / cells, allowing the entry of plasma proteins and cellular elements into the joint; and 2 / a chronic inflammatory phase occurring in the sub-synovium and the sub-chondral bone, characterized by pannus / granular tissue / articular formation, bone erosion and cartilage destruction. The pannus can form joints / adhesions and scar tissue, causing joint deformities typical of rheumatoid arthritis.

The disease of rheumatoid arthritis is not known. Infectious substances such as bacteria and viruses have been linked to this. According to current assumptions, Epstein-Barr virus / EBV / causes rheumatoid arthritis.

Currently the treatment of rheumatoid arthritis consists mainly of the needle _easing} nets using a non-steroidal anti-inflammatory drugs. Treatment with non-steroidal anti-inflammatory drugs is mainly effective in the early stages of rheumatoid arthritis; it is unlikely that inflammation of the joints will be eliminated if the disease has been present for more than a year. Gold, Methotrexate, Immunosuppressive ••• 4 ···

- 7 substances and corticosteroids have been tried with moderate success.

On the other hand, osteoarthritis is an inherited, non-inflammatory disorder of the movable joints characterized by deterioration and wear of the articular cartilage and the formation of new bone on the surface of the joints. As osteoarthritis progresses, the surface of the articular cartilage breaks down and worn parts are released into the joint fluid, which then stimulates phagocytosis / macrophages / macrophages. This eventually results in an inflammatory reaction in osteoarthritis.

Common clinical symptoms of osteoarthritis are cartilage and bone enlargement of the finger joints, initial stiffness and painful movement.

Common symptomatic treatments for osteoarthritis include painkillers, anti-inflammatory drugs, steroids and physical therapy.

Many phosphonic acid derivatives have been suggested for the treatment and prevention of disorders associated with abnormal calcium and phosphate metabolism. For example, numerous literature discloses compositions containing polyphosphonates, particularly diphosphonates, such as ethane-1-hydroxy-1,1-diphosphonic acid (EHDP), and their use to prevent and promote abnormal calcium and phosphate deposition in animals. Examples of such literature include U.S. Patent Nos. 3,683,080; 4,230,700; and 4,868,164. There are many other places ···

- 8 (Heterocyclic-Substituted Diphosphonic Acids which are useful in the treatment of osteoporosis and / or arthritis, such references being 5,071,840,

U.S. Patent Nos. 868,164, 5,104,863, 4,267,108, 4,746,654, and 4,876,247; European Patent Application Nos. 100,718, 170,228, 186,405, and 298,553; and U.S. Patent Nos. 4,754,993, 4,939,130 and 4,971,958; WO 90/12017 and WO 91/10646; and Australian Patent Applications AU-A-26738/88 and AU-A-45467/88. Finally, U.S. Pat. No. 4,208,401 discloses non-heterocyclic ring-substituted quaternary ammonium diphosphonates which are useful as anti-stone agents.

DE 40 11 777 discloses heterocyclic-substituted diphosphonates in which the heterocyclic ring may be substituted by lower alkyl. The heterocyclic ring is attached to the phosphonic acid group via a quaternary, non-ring nitrogen atom. The above German Patent also discloses that these compounds are specifically inhibitors of bone resorption and are thus useful in the treatment of osteoporosis, inflammatory and degenerative joint diseases, peridontitis and hyperparathyroidism / hyperthyroidism.

The compounds of the present invention have a bone protective effect on arthritic conditions at the site of joint deterioration, and this effect is an additional benefit in the treatment of arthritis over drugs that only reduce the inflammatory symptoms. The term "bone protective effect" as used herein means that the compounds have a disease modifying effect on the bone and surrounding soft tissue at the site of joint deterioration.

Surprisingly, it has been found that the compounds of the present invention wherein the nitrogen atom is quaternized have better activity against bone resorption and are more therapeutically useful in the treatment of osteoporosis, rheumatoid arthritis and osteoarthritis than the nitrogen-containing compounds which are not quaternized. The compounds of the invention also exhibit unusual solubility properties. Therefore, the compounds of the invention are more readily absorbed orally. The easier the compound is absorbed, the more effective it may be at lower doses. Lower doses are generally beneficial because they reduce unwanted side effects.

It is an object of the present invention to provide new, more potent compounds useful in the treatment of osteoporosis and anti-arthritis agents which are particularly useful in the treatment of osteoarthritis and rheumatoid arthritis.

It is a further object of the present invention to provide pharmaceutical compositions for the treatment of osteoporosis and arthritis / arthritis, in particular rheumatoid arthritis and osteoarthritis / osteoarthritis.

- 10 · * ♦ <· «* ·

Finally, it is an object of the present invention to provide methods of treating and preventing osteoporosis and arthritis, in particular rheumatoid arthritis and osteoarthritis.

The above and other objects of the invention will be apparent from the following detailed description of the invention.

The present invention relates to quaternary nitrogen-containing phosphonate derivatives and their pharmaceutically acceptable salts and esters, wherein:

m is an integer from 0 to 10;

n is an integer from 1 to 10;

and m + n = 1-10;

6 9 6 / R is nothing, -SR, -R SR, hydrogen, optionally, substituted with three C1-8 alkyl, -0R, ^C0 2 ^{R '-0} 2 ^CR' '^ ^N A, N / R ³ / C / O / R ^3, C / 0 / N / R _^3/2, halo3 atom, C / O / R is nitro, or hydroxy, optionally substituted saturated monocyclic or polycyclic heterocyclic ring or an optionally substituted saturated a monocyclic or polycyclic carbocyclic ring;

b / R ³ is -SR ³ , -R ^ SR ³ , hydrogen, optionally "3-substituted C 1 -C 8 alkyl, -OR,

-CO ₂ R ^3, -O2CR ³ N / R ^3/2, -N / R ³ / c / O / R ^3, C / O / N / R _^3/2, halogen, C / O / R ³ , nitro or hydroxy, optionally substituted saturated monocyclic or polycyclic heterocyclic ring, optionally substituted saturated monocyclic or polycyclic carbocyclic ring, optionally substituted unsaturated monocyclic or polycyclic heterocyclic, optionally monocyclic or polycyclic and combinations thereof.

, c / each R is optionally substituted C 1-35 alkyl, optionally substituted phenyl, benzyl or -R SR;

d / R H, optionally substituted C 1-8 -C 6 alkyl or -R SR;

e / Κθ hydrogen, C / O / R ^7, C / S / R ^7, C / O / N / R ⁷ / C / O / ^OR7, -C / S / N / R _^7/2 , -C / S / OR ⁷ , wherein R ^{7 is} hydrogen or optionally substituted C 1 -C 8 alkyl;

f / R is -COOH, -SO ₃ H, -PO ₃ H _2, or -P / O / OH / R ⁴ , e ⁴ R is C 1-3 alkyl;

g / R is optionally substituted C 1 -C 8 alkyl; and h / r8 is hydrogen, halogen, -SR4, -R4R5R6, amino or hydroxy or optionally substituted C1-C8 alkyl.

In the general formula (1), quaternary nitrogen · ·, ·

- 12 atoms must be linked to the carbon atom containing the phosphonic acid via a linking chain and must not be directly attached to the carbon atom bearing the phosphonic acid.

The present invention also provides pharmaceutical compositions comprising a safe and effective amount of a compound of the invention and pharmaceutically acceptable excipients.

Finally, the invention relates to methods for treating or preventing conditions in humans or other mammals characterized by abnormal calcium and phosphate metabolism such as osteoporosis and arthritis / arthritis / osteoarthritis / osteoarthritis inflammation/. The method of treatment comprises administering to a human or other mammal in need of treatment a safe and effective amount of a compound or composition of the invention.

The definitions of the terms used above are listed below.

The heteroatom is nitrogen, sulfur or oxygen. Groups containing one or more heteroatoms may contain different heteroatoms.

The alkyl group is an optionally substituted straight or branched, saturated or unsaturated hydrocarbon chain, if the hydrocarbon chain is saturated, it has from 1 to 8 carbon atoms, preferably unless otherwise stated, having from 1 to 4 carbon atoms, and if the hydrocarbon chain is unsaturated unless otherwise stated, contains from 2 to 4 carbon atoms. so

The term "alkyl" as used herein includes unsaturated alkenyl hydrocarbons having at least one olefinic double bond and unsaturated alkynyl hydrocarbons having at least one triple bond. Preferred alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, and butyl.

The carbocyclic ring or carbocyclic group as used herein refers to an optionally substituted, saturated, unsaturated or aromatic hydrocarbon ring. The carbocyclic ring may be monocyclic or polycyclic. Monocyclic rings generally contain from 3 to 8 atoms, preferably from 5 to 7 atoms. Two-ring polycyclic rings

6-16 atoms, preferably 10-12 atoms, and those consisting of three rings generally contain 13-17 atoms, preferably 14-15 atoms.

The heteroalkyl group is an optionally substituted saturated C 3 -C 8 chain containing carbon atoms and one or two heteroatoms.

The term heterocyclic ring or heterocyclic group as used herein denotes an optionally substituted, saturated, unsaturated or aromatic ring containing carbon atoms and one or more heteroatoms in the ring. Heterocyclic rings can be mono- or polycyclic rings.

Monocyclic rings generally contain from 3 to 8, preferably from 5 to 7, atoms. Polycyclic ring systems consisting of two rings generally contain from 6 to 16, preferably from 10 to 12, atoms.

Polycyclic ring systems consisting of three rings generally contain from 13 to 17, preferably from 14 to 15, atoms. A heterocyclic ring system may consist of heterocyclic or heterocyclic and carbocyclic rings. Each heterocyclic ring system must have at least one nitrogen atom. Unless otherwise stated, any other heteroatom is independently selected from the group consisting of nitrogen, sulfur and oxygen.

The aryl group is an aromatic carbocyclic ring. Preferred aryl groups include, but are not limited to, phenyl, tolyl, xylyl, cumyl and naphthyl.

The heteroaryl group is an aromatic heterocyclic group. Preferred heteroaryl groups include, but are not limited to, thienyl, furyl, pyrrolyl, pyridyl, pyrazinyl, oxazolyl, thiazolyl, quinolyl, pyrimidinyl, and tetrazolyl.

The alkoxy group is an oxygen atom having a hydrocarbon chain substituent, wherein the hydrocarbon chain is an alkyl or alkenyl group (e.g., -O-alkyl or -O-alkenyl). Preferred alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and alkyloxy groups.

The hydroxyalkyl group is a substituted hydrocarbon chain having a hydroxy substituent (e.g., -OH) and may also have other substituents. Preferred hydroxyalkyl groups include, but are not limited to, hydroxyethyl and hydroxypropyl.

The carboxyalkyl group is a substituted hydrocarbon chain having a carboxy substituent (e.g., -COOH).

- 15 • ·· and may have other substituents. Preferred carboxyalkyl groups are carboxymethyl and carboxyethyl and their acids and esters.

The aminoalkyl group is a hydrocarbon chain (e.g., an alkyl chain) substituted with an amino group (e.g., NH-alkyl), such as an aminomethyl group.

The alkylamino group is an amino group having one or two alkyl substituents, such as -N-alkyl, such as dimethylamino.

The alkenylamino group is an amino group having one or two alkenyl substituents (e.g., -N-alkenyl).

The alkynyl amino group is an amino group with one or two alkenyl substituents (e.g., -N-alkynyl).

The alkylimino group is an imino group with one or two alkyl substituents (e.g., -N-alkyl).

The arylalkyl group is an alkyl group substituted by a group. Preferred arylalkyl groups are benzyl and phenylethyl.

The arylamino group is an amino group substituted with an aryl group (e.g., -NH-aryl).

The aryloxy group is an oxygen atom with an aryl substituent (e.g., -O-aryl).

The acyl or carbonyl group is a carbon atom double bonded to an oxygen atom, for example R-C (= O). Preferred acyl groups include, but are not limited to, acetyl, propionyl, butanoyl, and benzoyl.

··· ··· ···

The acyloxy group is an oxygen atom with an acyl substituent (e.g., -O-acyl), such as -O-C (= O) -alkyl.

The acylamino group is an amino group with an acyl substituent (e.g., -N-acyl), such as -NH-C (= O) -alkyl.

The halo or halo group is chloro, bromo, fluoro or iodo. Preferred halogen groups are chlorine, bromine and fluorine.

The lower hydrocarbon group mentioned herein, for example lower alkyl, unless otherwise specified, contains from 1 to 6, preferably from 1 to 4, carbon atoms.

g In the specification, the thio substituent (-SR or -R SR) also includes thiols (-SH),

67 when R = H; thioesters f-SC / O / R j when R = -C / O / R;

67 the dithioesters are f-SC / S / RJ when R = C / S / R; the thiocarbamethers are -SC / O / N / R / ₂ J when R = C / O / N / R / ₂ ; dithiocarbamates -SC £ / S / N / R ^7/7 ^> where ^R6 = C / S / N / R _^7/2; thiocarbonates 7 67 = SC / O / OR J when R = C / O / OR; and dithiocarbonates

6 77 £ -SC / S / OR 7 when R = C / S / OR. R is generally hydrogen or C 1-8 alkyl. Any SR 1 substituent may itself be substituted with an R group, i.e.

6 9. , r SR, wherein R is optionally substituted with 1 to 8 senatenes

6 alkyl groups. Thus, other thio substituents designated R SR include alkyl thiols, alkyl thioesters, alkyl dithioesters, alkyl thiocarbamates, alkyl dithiocarbamates, alkyl thiocarbonates, and alkyl dithiocarbonates.

The term "diphosphonate or diphosphonic acid" is used herein to refer to those phosphonate or phosphonic acid derivatives.

- 17, in which two phosphonate groups are attached to the same carbon atom and are used interchangeably with the terms diphosphonates and diphosphonic acids. Using this structure, R is po ₃ h ₂ .

As used herein, the term "phosphonic acid carbon" refers to the carbon atom to which the phosphonic acid group is attached. When another phosphonic acid group is attached to this carbon atom, the compound is a diphosphonate. When a sulfonate group is attached to the above carbon atom, the compound is a phosphonosulfonate. When a carboxylate group is attached to said carbon atom, the compound is phosphonocarboxylate. When a phosphinic acid group is attached to this carbon atom, the compound is phosphonoalkylphosphinate.

A pharmaceutically acceptable salt is a cationic salt formed on an acidic (e.g. carboxyl) group; or an anionic salt formed on a basic (e.g. amino) group. Many such salts are known in the art, for example from International Publication No. 87/05297. Preferred cationic salts are the alkali metal salts (such as the sodium and potassium salts) and the alkaline earth metal salts (such as the magnesium and calcium salts). Preferred anionic salts are halides (e.g. chlorides), acetates and phosphates.

The biohydrolyzable ester is an ester of a quaternary nitrogen-containing heterocyclic phosphonate that does not impair the therapeutic effect of the compounds or is readily metabolised by humans or other mammals. Many such esters are known in the literature, for example, · <·

• · · · · ♦ ····· ·· · ···

- International Publication Number 87/05297. Such esters include lower alkyl esters, lower acyloxyalkyl esters such as acetoxymethyl, acetoxyethyl, carbamoyloxymethyl, pivaloyloxymethyl and pivaloyloxyethyl esters. , lactonyl esters such as phthalidyl and thiophthalidyl esters, lower alkoxy acyloxyalkyl esters such as methoxycarbonyloxymethyl, ethoxycarbonyloxyethyl and isopropoxycarbonyl. oxyethyl esters, alkoxyalkyl esters, choline esters and acylaminoalkyl esters, such as acetamidomethyl esters.

As mentioned above, and as described herein, substituent groups may themselves be substituted. This substitution may take place with one or more substituents. Possible substituents include, but are not limited to, C. Hansch and Leo A., Substituent Constans for Correlation Analysis in Chemistry and Biology, 1979 / c. It works. Preferred substituents include, but are not limited to, alkyl, alkenyl, alkoxy, hydroxy, oxo, amino, aminoalkyl (e.g., aminomethyl, etc.), cyano, halo, carboxy, alkoxy. acetyl (e.g. carboethoxy, etc.), thio, thiol, aryl, cycloalkyl, heteroaryl, heterocycloalkyl (e.g., piperidyl, morpholinyl, piperazinyl, pyrrolidinyl, etc.), imino, thioxo , hydroxyalkyl, aryloxy, arylalkyl and combinations thereof.

This invention relates to novel quaternary nitrogen-containing phosphonate compounds and their pharmaceutically acceptable salts and esters containing quaternary nitrogen. The quaternary nitrogen atom is phosphonic acid • · «

- It is linked to 19 carbon atoms through a linking chain.

The carbon atom to which the phosphonic acid group is attached may be unsubstituted (i.e., hydrogen) or substituted. Phosphonic acid carbon can carry two phosphonate groups to form a diphosphonate derivative; a phosphonate and a carboxylate moiety, such as a phosphonocarboxylate derivative; a phosphonate group and a sulfonate group such as a phosphonosulfonate derivative; a phosphinate group and a phosphonate group such as a phosphonoalkyl phosphinate derivative.

The phosphonate compounds of the invention containing the quaternary nitrogen atom and their pharmaceutically acceptable salts and esters have the formula (I).

It has many meanings in this general formula

9 6 non-cyclic group may thus nothing, -SR, -R SR, hydrogen 3 3 3 H, C1-8 alkyl, -OR, -CO ₂ R, -O ₂ CR, N / R ^3/2, N / R ³ / C / O / R ^3, C / O / N / R ^3/2, halogen, C / QLT ?, nitro, hydroxy, optionally substituted saturated monocyclic or polycyclic heterocyclic ring, and optionally substituted, saturated monocyclic or polycyclic carbocyclic ring.

In the above general formula, the quaternary nitrogen atom is attached to the carbon atom bearing the phosphonic acid via a linking chain. In the general formula, n, which is an integer from 1 to 10, represents the length of the linking chain.

• · · · · ·····································•

- 20 These connecting links are composed of different R ³ groups. R ³ is selected from -SR ^, R ^ SR ^, 3 3 is hydrogen, C1-8 alkyl, -0R, -CC> R _2, -O ₂ CR ^3, N / R ^3/2, -N / R ³ / C / O / R ^3, C / 0 / N / R ^3/2, halogen,

-C / O / R, nitro, hydrogen, an optionally substituted saturated monocyclic or polycyclic heterocyclic ring, optionally substituted, saturated monocyclic or polycyclic carbocyclic ring, optionally substituted, unsaturated monocyclic or polycyclic heterocyclic, optionally substituted polycyclic carbocyclic ring and combinations thereof.

Finally, phosphonate compounds of the invention containing a quaternary nitrogen atom R can be -COOH, -SO ^ H, -PC> ₃ H _2, or P / 0 // 0H / R ^4, where R ⁴ -C 1-8 al ~ - 4 alkyl . A preferred group R is -PO ^ H. es a -P / O // OH / R.

g r is a substituent on a carbon atom bearing a phosphonic acid, denotes hydrogen, halogen, -SR, -R SR °, amino or hydroxy, or an optionally substituted C 1-8 alkyl group. Preferred R groups are hydroxy, halo and amino. R is optionally substituted C 1 -C 35 alkyl, optionally substituted phenyl 9 -C 2 or benzyl or -R SR. Preferred R groups are given

- 9 6-substituted-C 1 -C 8 alkyl and -R SR groups.

Preferred quaternary nitrogen-containing phosphonates of the above R ³ groups are N- (5-hydroxy-3,3-diphosphonopropyl) -N, N-dimethyl-N-21-pentylammonium (11). iodide; N- (4-hydroxy-4,4-diphosphonobutyl) -N, N, N-trimethylammonium chloride (III); and N- (3-hydroxy-3,3-diphosphonopropyl) -N, N, N-triylethylammonium chloride (TV).

In the formula (I), R ¹ can also be a saturated monocyclic or polycyclic heterocyclic group.

Preferred phosphonates containing a quaternary nitrogen atom which contains a monocyclic or polycyclic heterocyclic group as R 4 and in which the quaternary nitrogen atom is bonded to a carbon atom bearing the phosphonic acid are represented by N- [3-hydroxy-3 diphosphonopropyl / -N, N-dimethyl-N- / 2-methyl-piperidine / ammonium chloride.

Further preferred are the compounds of formula (VI) and (VII) of the invention.

Further preferred compounds of the invention are thio-substituted quaternary nitrogen-containing phosphonates of the formulas VIII, IX, X and XI.

Specific examples of compounds of the invention include:

N- / 4-hydroxy-4,4-diphosphono-butyl / N, N, N-trimethyl-ammonium iodide

N- / 3-hydroxy-3,3-diphosphonopropyl / -N, N-dimethyl-N-pentyl ammonium iodide;

N- / 3-hydroxy-3,3-diphosphonopropyl / N, N, N-trimethyl ammonium iodide;

- 22 ··· ··· * · · · «» · «« · ·· ·

N-cycloheptyl-N, N-dimethyl-N- / diphosphonopropyl meti1 / ammonium iodide;

N- / 2-acetylthio-ethyl / -N- / 4-hydroxy-4,4-diphosphono-butyl / -N, N-dimethyl ammonium bromide;

N- / 2-acetylthio-ethyl / -N- / 3-hydroxy-3,3-diphosphonopropyl / -N-methyl-N-pentyl ammonium bromide;

N- / 4-hydroxy-4,4-diphosphono-butyl / -N- / 3-mercapto-propyl / N, N-dimethyl ammonium chloride;

N- / 4-hydroxy-4,4-diphosphono-butyl / -N- / mercaptomethyl / -N, N-dimethyl ammonium chloride;

N- (4-hydroxy-4,4-diphosphono-butyl) -N- (4-methoxybutyl) -N, N-dimethylammonium chloride;

N- / 4-hydroxy-2-mercapto-4,4-diphosphono-butyl / N, N, N-trimethylammonium chloride;

N- / 4-hydroxy-2-acetylthio-4,4-diphosphono-butyl / N, N, N-trimethylammonium chloride;

N- / 3-hydroxy-2-mercapto-3,3-diphosphonopropyl / -N, N-dimethyl-N-pentyl ammonium chloride;

N- / 3-hydroxy-2-acetylthio-3,3-diphosphonopropyl / -N, N-dimethyl-N-pentyl ammonium chloride;

N- / 3-hydroxy-3,3-diphosphonopropyl / -N-methyl-N-pentyl-N- £ 2- / 3-pyridyl / ethyl / ammonium chloride;

N-cycloheptyl-N- / 2-mercapto-ethyl / -N-methyl-N- / diphosphonomethyl-/ ammonium chloride;

N-cycloheptyl-N / mercaptomethyl / -N-methyl-N- / diphosphonomethyl-/ ammonium chloride;

N, N-dimethyl-N- (4,4-diphosphonobutyl) -N- (2- [3-piperidinyl] ethyl] -monium ammonium chloride.

·· "

For the purpose of determining and evaluating the pharmacological action, the phosphonate compounds are assayed in animals using various assays well known in the art. Thus, the in vivo action against bone resorption is conveniently determined by assaying the ability of these compounds to inhibit bone resorption characterized by abnormal calcium and phosphate metabolism. Such a test is known in the art as the Schenk model. Another useful assay known in the art is the adjuvant arthritis test. The in vitro hydroxyapatite crystal growth inhibition assay can also be used. These and other suitable assays for pharmacological activity can be found in the literature, for example, Shinoda et al., Calcified Tissue International, 35, 87-99 (1983); Schenk et al. , Calcified Tissue Research, 11, 196-214 (1973); Russell et al., Calcified Tissue Research 6: 183-196 (1970); Muhlbauer and Fleisch, Mineral Electrolyte Metab., 5, 296-303 (1981); Nancollas et al., Órai Bioi., 15, 731 (1970); U.S. Patent Nos. 3,683,080; and 4,134,969; and European Patent Application EP 189,662. Some of the above tests for pharmacological activity are described in detail in the following examples.

In addition to being useful in the treatment and prevention of abnormal calcium or phosphate metabolic conditions, the compounds of the present invention are also useful in treating and preventing disorders associated with abnormal calcium or phosphate metabolism. · 4 · •• 4 ♦ · · ··· ···

- There are 24 areas of application. For example, the compounds of the invention are likely to be used as bone scanners after labeling with 99 µm technetium. The compounds of the present invention may also be used as chelating / complexing agents for polyvalent metal ions, especially divalent (e.g. calcium and magnesium) and trivalent (e.g. indium) metal ions. Thus, the compounds of the present invention can be used as builders in detergents or for treating / softening water. The compounds may also be used as stabilizers for compounds. In addition, they can be used to prevent stones (such as gall, kidney stones, etc.) and / or to prevent the formation of tartar on the teeth. Finally, the compounds of the invention can be used as non-toxic herbicides for animals.

Quaternary nitrogen-containing phosphonates for use in the pharmaceutical compositions of the present invention are described, without limitation, in Examples 1-5. can be prepared according to the example of.

Pharmaceutical preparations containing novel phosphonates containing quaternary nitrogen

The novel quaternary nitrogen-containing phosphonate compounds of the present invention may be administered to humans or other mammals in a variety of ways including, but not limited to, oral dosage forms or injections such as intravenous, intramuscular, intraperitoneal and subcutaneous injection. The novel quaternary nitrogen according to the invention.

Many other dosage forms containing phosphonate compounds having 25 atoms can be readily prepared by one of ordinary skill in the art using the appropriate pharmaceutical excipients / excipients. Oral dosage forms are generally the most preferred, according to the wishes of the patients.

As used herein, the term "pharmaceutical composition" refers to combinations containing a safe and effective amount of a quaternary nitrogen-containing phosphonate compound or a mixture thereof with pharmaceutically acceptable excipients.

As used herein, the term "safe and effective amount" means an amount of a compound or composition that is large enough to significantly and positively modify the symptoms and / or condition to be treated, but small enough to avoid more serious side effects / acceptable benefits. / risk ratio / according to reasonable medical judgment. The safe and effective amount of the active ingredients used in the pharmaceutical compositions of the present invention will depend on the particular condition being treated, the age and physical condition of the patient being treated, the severity of the condition, the nature of the concurrent treatment, the particular active ingredient used. pharmaceutically acceptable excipients and similar factors known and experienced by a physician.

The term "pharmaceutically acceptable excipient" refers to an inert, pharmaceutically inactive substance known to those skilled in the art, which is compatible with the specific quaternary nitrogen-containing phosphonate compound selected for use. Pharmaceutically acceptable binders / excipients / - but are not limited to polymers, resins, plasticizers, fillers, binders, lubricants, lubricants, disintegrants / solvents, co-solvents, buffer systems, surfactants, preservatives, sweeteners, pharmaceutical grade dyes and pigments; and viscosity regulators.

The term oral dosage form as used herein refers to any pharmaceutical composition which is administered systemically to a subject by oral administration to the subject's gastrointestinal tract. For purposes of the present invention, the dosage unit dosage form may be a tablet, coated or uncoated, solution, suspension or capsule, coated or uncoated.

The term "injection" as used herein refers to a pharmaceutical composition that is systemically administered to a human or other mammal by administering a solution or emulsion containing the active ingredient through the skin of an individual to deliver the solution or emulsion to a subject's intravenous, intramuscular, or by subcutaneous injection.

The extent of systemic administration is well within the reach of one skilled in the art

- 27 can control it by influencing one or more of the following:

the active ingredient;

b / pharmaceutically acceptable excipients, insofar as the changes do not affect the action of the selected specific active ingredient;

c / the type of binder and the required hardness and swelling properties of the binder;

d / the time-dependent states of the binder itself and / or the time-dependent states in the binders;

e / particle size of the granulated active ingredient; and f / pH dependent states of the binders.

In particular, the solubility, acidity and sensitivity of the various non-ring-containing phosphonate phosphonate agents can be used as a guideline for their hydrolysis. Thus, acid addition salts, carboxylic acid salts such as alkali metal salts, alkaline earth metal salts and the like can be used. and esters such as alkyl, aryl, aralkyl esters. In addition, oral dosage forms may provide appropriate pH conditions by adding a suitable buffer to the active ingredient, according to the desired mode of release.

As noted above, pharmaceutically acceptable binders / excipients /, but not limited to, resins, fillers, binders, lubricants, solvents, lubricants, disintegrants, co-solvents, surfactants, preservatives, · · ··· ··· · · · · · · ·

- sweeteners, excipients, buffer systems, pharmaceutical grade dyes or pigments and viscosity regulators.

The preferred solvent is water.

Materials useful in the present case are described in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, 1990, p. 1288-1300. The pharmaceutical compositions useful in the present case generally contain from 0% to 2% of the raw material.

Dyes and pigments useful in the present invention are described in Hphi adbook of Pharmaceutical Excipients, p. 81-90, 1986 by the American Pharmaceutical Association and the Pharmaceutical Society of Great Britain. The pharmaceutical compositions of the present invention generally contain from 0 to 2% of a dye or pigment.

Preferred co-solvents include, but are not limited to, ethanol, glycerol, propylene glycol, and polyethylene glycols. The pharmaceutical compositions of the invention contain 0-50% co-solvent.

Preferred buffer systems include, but are not limited to, acetic acid, boric acid, carbonic acid, phosphoric acid, succinic acid, maleic acid, tartaric acid, citric acid, benzoic acid, lactic acid, glyceric acid, gluconic acid, glutaric acid and glutamic acid and their sodium, potassium and ammonium. Particularly preferred are phosphoric acid, tartaric acid, citric acid, acetic acid and salts thereof. The pharmaceutical compositions of the present invention generally contain from 0 to 5% buffer.

Preferred surfactants include, but are not limited to, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene monoalkyl ethers, sucrose monoesters, and lanolin esters and ethers. , sodium, potassium and ammonium salts of alkyl sulfate salts and fatty acids. The pharmaceutical compositions of the present invention contain from 0 to 2% surfactant.

Preferred preservatives include, but are not limited to, alkyl esters of phenol, para-hydroxybenzoic acid, o-phenylphenol, benzoic acid and its salts, boric acid and its salts, sorbic acid and its salts, chlorobutanol, benzyl alcohol, thimerosal, phenylmercury acetate and nitrate, nitromerzole, benzalkonium chloride, cetylpyridinium chloride, methyl paraben and propyl paraben. Particularly preferred are the salts of benzoic acid, cetylpyridinium chloride, methyl and propyl paraben. The compositions of the invention generally contain from 0% to 2% of a preservative.

Preferred sweeteners include, but are not limited to, sucrose, glucose, saccharin, sorbitol, mannitol, and aspartame. Sucrose and saccharin are particularly preferred. The pharmaceutical compositions of the present invention contain 0-5% sweetener.

Preferred viscosity control agents include methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium alginate, carbomer, povidone, acacia, guar gum, xanthan gum and tragacanth. Particularly preferred are methylcellulose, carbomer, xanthan gum, guar gum, povidone, sodium carboxymethylcellulose and magnesium aluminum silicate. The compositions of the present invention contain 0-5% of a viscosity regulator.

• ·

Preferred excipients include, but are not limited to, lactose, mannitol, sorbitol, calcium phosphate (three bases), calcium phosphate (dibasic), compressible sugar, starch, calcium sulfate, dextrose, and microcrystalline cellulose. The compositions of the present invention contain from 0% to 75% filler.

Preferred lubricants / lubricants / materials include, but are not limited to, magnesium stearate, stearic acid and talc. The pharmaceutical compositions of the present invention contain 0.5 to 2% of a lubricant.

Preferred lubricants include, but are not limited to, talc and colloidal silica. The compositions of the present invention contain from 1% to 5% of a lubricant.

Preferred disintegrants / disintegrants include, but are not limited to, starch, sodium starch glycolate, crospovidone, croscarmelose sodium and microcrystalline cellulose. The pharmaceutical compositions of the present invention contain from 4% to 15% of a disintegrant.

Preferred binders include, but are not limited to, gum arabic, tragacanth, hydroxypropylcellulose, pregelatinized starch, gelatin, povidone, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, sugar solutions such as sucrose and sorbitol solution and ethyl cellulose. The compositions of the invention contain from 1% to 10% of a binder.

The compounds of the present invention may comprise from about 0.1% to about 99.9% by weight of the compositions of the invention.

··· • t · · · · · · · · · · · · · · · · · · ···

The compounds of the present invention preferably comprise from about 20% to about 80% by weight of the compositions of the invention.

Thus, the compositions of the present invention comprise 15-95% of a quaternary nitrogen-containing phosphonate compound, or mixtures thereof, 0-2% of a raw material, 0-50% of a co-solvent, 0-5% of a buffer system, 0-2% of a surfactant, 0-2% of a preservative. 0-5% sweetener, 0-5% viscosity regulator, 0-75% filler, 0.5-2% lubricant, 1-5% lubricant, 4-15% disintegrant and 1-10% binder ·

Suitable pharmaceutical compositions are described in FIGS. Examples are described in Examples. One of ordinary skill in the art will know to vary these non-limiting examples for the preparation of a wide variety of pharmaceutical formulations.

The choice of pharmaceutical excipients / excipients for use with the quaternary nitrogen-containing phosphonate compounds of the present invention is essentially determined by the manner in which the phosphonate is administered. If the compound is to be injected, the preferred pharmaceutical carrier is sterile saline, the pH of which is adjusted to about 7.4. However, the preferred mode of administration of the phosphonates of the present invention is oral administration, and the preferred unit dosage form is therefore tablets, capsules, and the like, containing from about 0.1 to about 600 mg of the phosphonic acid compound. For unit dosage forms suitable for oral administration • · ···

32 suitable pharmaceutical carriers are well known in the art. The choice of these depends on secondary considerations, such as taste, cost and storage, which are not critical to the objectives of the invention, and will be readily accomplished by those skilled in the art.

The term mg P as used herein is the weight of a phosphorus atom present in a certain amount of the phosphonic acid compound of the invention. This unit is used to standardize the amount of phosphonic acid compounds present in the pharmaceutical compositions and methods of the invention. For example, N- (4-hydroxy-4,4-diphosphonobutyl) -N, N-dimethyl-N- (2-mercaptoethyl) -ammonium chloride has a molecular weight of 373.5 g / mol, trans (62 g / mol) is the two phosphorus atoms present in the molecule. Thus, 1 mg of this compound contains 0.17 mg of P. Thus, to contain 0.17 mg of P in the pharmaceutical composition, the composition must contain 1 mg of the compound; and to administer 0.17 mg P / kg of compound to a 50 kg patient, 50 mg of compound is required.

The pharmaceutically acceptable carrier used in connection with the diphosphonates of the present invention is used in a concentration sufficient to provide a suitable particle size ratio. Pharmaceutically acceptable carriers are preferably about 20-80% by weight of the total composition.

- 33 Methods for the Treatment and Prevention of Disorders Associated with Abnormal Calcium and Phosphate Metabolism

The invention also relates to methods of treating and preventing disorders associated with abnormal calcium and phosphate metabolism. These methods comprise administering to a human or inferior animal in need of such treatment a safe and effective amount of a phosphonate compound of the invention.

The preferred mode of administration is oral, but other known modes of administration may be contemplated, such as transmucosal (dermatomucosally), transdermal, rectal, etc., and parenteral, such as subcutaneous, intramuscular, intra-articular, injection, and the like. Administration may also be by inhalation. Specific routes of administration include, but are not limited to, oral, transdermal, mucosal, sublingual, intramuscular, intravenous, intraperitoneal, and subcutaneous, and topical applications.

As used herein, abnormal calcium and phosphate metabolism refers to conditions 1 / which are characterized by abnormal movement of calcium and phosphate leading to general and specific bone loss or excessive levels of calcium and phosphate in body fluids; and 2 / which are characterized by causing or resulting from abnormal calcium and phosphate deposition in the body. The first category includes, but is not limited to, the * ·

- 34 osteoporosis, Paget's disease, hyperthyroidism, malignant high blood calcium mirror, heterotopic / abnormal bone formation and bone dissolution bone metastases. The second category includes, but is not limited to, progressive osteoarthritis with osteoporosis, general calcinosis (limescale deposition in various tissues), and diseases such as arthritis / arthritis, inflammation of the nerves, which tend to cause the tissue in question to deposit calcium and phosphate.

As used herein, the term rheumatoid arthritis is an unknown pathogenic, chronic, generalized and arthritic disease. This disease is characterized by deterioration of articular cartilage, ligaments, tendons and bone.

The term osteoarthritis refers to a non-inflammatory disorder of the moving joints. This is characterized by the deterioration and wear of the articular cartilage and the formation of new bone on the joint surface.

The terms "person at risk" and "person in need of such treatment" refer to a human or inferior animal in whom there is a significant risk due to abnormal calcium and phosphate metabolism if left untreated, and to a human or inferior animal in which abnormal calcium and phosphate metabolism have been diagnosed. Such persons are, for example, women after the menopause, on certain steroid treatments. · · ·································• ·· 4 «· ···

- 35 patients, persons treated with anticonvulsants, persons diagnosed with Paget's disease, patients with parathyroid hypertrophy, persons with malignant elevated calcium calculus or bone metastases; persons suffering from one or more different forms of osteoporosis; persons in the population known to have a significantly higher than average chance of developing osteoporosis, such as post-menopausal women, men over 65 and persons treated with osteoporosis as a side effect, persons with advanced osteoporosis muscle inflammation or generalized calcinosis; and persons having arthritis, osteoarthritis, neuritis, mucosal inflammation, inflammation, or other inflammatory disease which predisposes the tissue to deposition of calcium and phosphate.

As used herein, the term "safe and effective amount" means an amount of the compositions or compounds of the invention that is large enough to significantly alter the condition to be treated, but small enough to balance the serious side effects / reasonable benefits / risks. should be avoided. The safe and effective amount of the phosphonate compounds of the present invention will depend on the particular condition being treated, the age and physical condition of the patient being treated, the severity of the condition, the duration of treatment, · * ·· «« · · · * ·

- the nature of the concomitant therapy, the specific phosphonate used, the specific pharmaceutically acceptable carrier employed, and similar factors known and experienced by the physician. However, a single dose may be 0.01 mg P - 3500 mg P or 0.0002 - 70 mg P / kg body weight (based on 50 kg body weight). A preferred single dose is 1 mg P to 600 mg P or 0.02 to 12 g P / kg body weight based on 50 kg body weight. Up to four such doses may be administered daily. A daily dose greater than 500 mg P / kg / day is not required to achieve the desired effect and may result in unwanted side effects. Higher doses in the above range are, of course, required for oral administration due to limited absorption.

Preferred embodiments of the invention are illustrated by the following examples. The examples are by way of illustration only and are not intended to limit the scope of the invention in any way, since many variations thereof are possible without departing from the spirit and scope of the invention.

1 ^ __ Geld

Synthesis of N- (4-Hydroxy-4,4-diphosphonobutyl) -N, N, N-trimethylammonium iodide

I. Synthesis of 4-N, N-N-Dimethylamino-1-hydroxybutylidene-diphosphonic acid

Containing 2.9 moles of 4- (N, N-dimethylamino) -butanoic acid, 2.0 mmol of phosphorus trichloride and 12 mmol of diethyl phosphite

The solution was stirred for 30 minutes at room temperature and kept at 60 ° C for 24 hours. The reaction mixture was cooled to room temperature and concentrated hydrochloric acid (50 mL) was added. The reaction mixture was heated and refluxed for 24 hours, cooled to room temperature, filtered through celite and the filtrate evaporated in vacuo. The crude product was triturated with ethanol, filtered and dried in vacuo.

II. Synthesis of N- (4-Hydroxy-4,4-diphosphonobutyl) -N, N, N-trimethylammonium iodide

The above diphosphonic acid (0.30 mmol) was dissolved in a mixture of water (10 mL) and ethanol (15 mL) and the pH of the solution was adjusted to 7.0 by addition of 1 N sodium hydroxide solution. Methyl iodide (1.50 mmol) was added to the reaction mixture and the mixture was refluxed for 24 hours. The mixture was then cooled and concentrated under reduced pressure. The solid residue is dissolved in a minimum amount of water and the quaternized product is precipitated by addition of isopropanol. The product was filtered off, rinsed with acetone and dried in vacuo.

2 _r _2Give it

Synthesis of N- (3-Hydroxy-3,3-diphosphonopropyl) -N, N-dimethyl-N-pentylammonium iodide

I. Synthesis of? 3-N, N-Dimethylamino-propylidene-7-diphosphonic acid

Essentially as described in Example 1, Part I

- 38 · 4 ····························································· · · · · · · · · / 3- / N, N-dimethylaminopyridine / -propilidén7-difoszfonsavvá.

II. Synthesis of N- (3-Hydroxy-3,3-diphosphonopropyl) -N, N-dimethyl-N-pentylammonium iodide

0.50 mmol of the above diphosphonic acid is dissolved in a mixture of 15 ml of water and 20 ml of acetonitrile and the pH of the solution is adjusted to 7.0 by the addition of 1 N sodium hydroxide solution. To the mixture was added pentyl iodide (2.50 mmol) and heated at reflux for 22 hours. The reaction mixture was then concentrated under reduced pressure and the solid product was triturated with acetone. The product can be recrystallized from a mixture of water and ethanol.

3 _r __example

Synthesis of N- (3-Hydroxy-3,3-diphosphonopropyl) -N, N, N-trimethylammonium iodide

Essentially, Example 1, Example II. . 3- (N, N-Dimethylamino) -propylidene-diphosphonic acid prepared according to Example 2, Part I is converted to N- (3-hydroxy-3,3-diphosphonopropyl) -N, N-trimethyl ammonium iodide.

4 _r __example

N- (2-Acetylthioethyl) -N- (4-hydroxy-4,4-diphosphonobutyl) -N, N-dimethylammonium bromide

0.75 mmol of 4- (N, N-dimethylamino) -1-hydroxybutylidene-diphosphonic acid (0.75 mmol) prepared in Part I of Example 1 was dissolved in a mixture of water (50 mL) and acetonitrile (35 mL). To this solution was added 3.75 mmol of S-acetyl-2-bromoethane thiol and the reaction mixture was heated at reflux for 12 hours. The reaction mixture was then concentrated under reduced pressure and the solid product was triturated with acetone. The quaternized product can be recrystallized from a mixture of water and ethanol.

5 _r __Enter

Synthesis of N- (2-Acetylthioethyl) -N- (3-hydroxy-3,3-diphosphonopropyl) -N-methyl-N-pentylammonium bromide

I. E -3- (N-Methyl-N-pentylamino) -propylidene-diphosphonic acid

Essentially, as described in Example 1, Part I, 3- (N-methyl-N-pentylamino) -propionic acid is converted to 3- (N-methyl-N-pentylamino) -propylidene diphosphonic acid. .

II. Synthesis of N- (2-Acetylthioethyl) -N- (3-hydroxy-3,3-diphosphonopropyl) -N-methyl-N-pentylammonium bromide

Substantially as described in Example 4, N-3-N-methyl-N-pentylamino-propylidene-diphosphonic acid was converted to N- (2-acetylthioethyl) -N- (3-hydroxy-3,3-). diphosphonopropyl / -N-methyl-N-pentyl ammonium bromide.

6 _r __example

Schenk model

Compounds are evaluated for their inhibitory activity against in vivo bone resorption and mineralization in an animal model system known as the Schenk model in bone metabolism. The basic principles of this method are Shinoda et al., FCalcif. Tissue Int. 35, 87-99 / 1983/7 and Schenk et al. Tissue Slit. 11, 196-214 / 19 73 / J.

Materials and procedures

Animals

Previously weaned, 17 days / 30g / h

Sprague Dawley rats (Charles River Breeding Laboratories) were transported with their mothers and placed in plastic cages with their mothers on arrival. At 19 days of age, the small animals received rat food and water as desired and were randomly assigned to treatment or control groups, each containing seven animals.

On day 1, then again on day 7, all animals were administered intraperitoneal / IP / injection of calcein / 1% solution in 0.9% saline; dose is 0.2 ml / 100 g body weight /.

On day 4, all animals were administered IP injection of tetracycline hydrochloride / 1% solution in 0.9% saline;

dose is 0.2 ml / 100 g body weight /. These compounds are effective in signaling bone and cartilage mineralization.

Dosage solutions and method of administration

For subcutaneous injection, each solution was prepared in 0.9% normal saline and adjusted to pH 7.4 with sodium hydroxide and / or hydrochloric acid. Dosage solutions were calculated by taking into account the powder weight of the active ingredient / mg molecular weight and hydration / mg / kg body weight, which corresponds to a. mg P / kg. Concentrations were calculated for a dose of 0.2 ml / 100 g body weight. Specifically, all compounds were administered at doses of 0.01, 0.1, 1.0 and 10.0 mg P / kg / day for 7 days. Compounds that exhibited activity at a dose of 0.1 mg P / kg / day were then assayed by logarithmic decrements up to 0.001 mg P / kg / day. Doses were adjusted daily based on changes in body weight.

Autopsy. Histological examination and histology

All animals were sacrificed by IP overdose of pentabarbitol on day 8 after dosing commenced. The tibia / sternum was prepared and placed in 70% ethyl alcohol. A tibial was gradually dehydrated in ethanol solutions and placed in methyl methacrylate as described by Schenk in Methods of Calcified Tissue Preparation / G.R. Dickson, Editor; Elsevier Science Publ., The Netherlands, 1984 //. The tibia was cut lengthwise through the metaphyseal section. One surface of the preparations was stained with silver nitrate and placed on microscope slides and evaluated with Quantimet Image Analyzer / Cambridge Instruments ··· ·

Inc./, both in incandescent and ultraviolet light.

Metaphyseal trabecular bone content was measured between the fluorescent label and the grown bone plate as a percentage of total area / bone + bone marrow. The width of epiphyseal bone plate growth is averaged from 10 measurements over the profile at equal distances.

Statistical evaluation of data by parametric and non-parametric analysis of variance and Wilcoxon classifier summation! test was used to determine a statistically significant effect compared to control animals. The Schenk model provides data on the inhibitory effect of compounds on bone resorption in vivo.

7 .___ Geld

Adjuvant arthritis model

There are several animal models of arthritis, including adjuvant-induced arthritis using Mycobacterium butyricum. This model mimics human rheumatoid arthritis in various ways (joint, swelling, interconnected spaces between cells by penetration of cells and pannus, bone resorption and entry of chemotaxis factors and lysosomal components into the joint space / / 1,2). Numerous prevention and therapeutic studies have reported the effective use of anti-inflammatory drugs (3,4) and diphosphonates (5,6) in arthritis.

References

1. Pearson, C., Wood F., Studies in Polyarthritis and

Other Lesions Induced by Injection of Mycobacterial

Adjuvant. 1. Generic Clinical and Pathological Characteristics and Somé Modifying Factors, Arth.

Rheum., 2, 440-459 (1959);

2. Blackman, A., Burns, J.W., Framer, J.B., Radziwonik,

H., Westwick, J., An X-ray Analysis of Adjuvant

Arthritis in the Rat. The Effect of Prednisolone and

Indomethacin, Agents and Actions, T_, 145-151 (1977);

3. Winter, C.A., Nuss, G.W., Adjuvant Treatment

Arthritis in Rats with Anti-inflammatory Drugs, Arth. Rheum., 9, 394-404 (1966);

4. Winder, C.V., Lembke, L.A., Stephens, M.D., Comparative Bioassay of Drugs in Adjuvant-Induced Arthritis.

Rats: Flufenamic Acid, Mefenamic Acid, and Phenylbutazone, Arth. Rheum., 12, 472-482 (1969);

5. Francis, M.D., Flora, L. King, W.R., The Effects of Disodium Ethane-1-Hydroxy-1-Diphosphonate on Adjuvant

Induced Arthritis in Rats, Calcif. Tiss. Slot, 9_,

109-121 (1972);

6. Flora, L., Comparative Anti-Inflammatory and Bone Protective Effects of Two Diphosphonates in Adjuvant Arthritis,

Arth. Rheum, 22, 340-346 (1979).

Adjuvant arthritis is a severe, cellulitis / cellular / connective tissue inflammation / and synovitis / arthritis / induced in male rats / Sprague Dawley or Lewis strain / caused by a single mineral of Mycobacterium butyricum ♦ ··· • SC / by injection / 8 mg / ml_ / day 0.

The compounds are administered once daily orally IV or parenterally (SC) and assays are performed prophylactically (from day 0) or therapeutic (day 9, 10 or 14) / protocol. Efficacy against arthritis can be measured by foot volume, weight loss, bone loss, or reactive new bone formation compared to saline-treated arthritis control animals. Treatment may be stopped and the inflammatory flare reaction may be tested, indicating the ability of a compound to maintain efficacy.

Materials and procedures

A. Animals

Animals were male Lewis rats (LEW). When the rats arrived, were they computer controlled _? randomly distributed according to numbers and placed in individual suspended wire cages. Food and water can be taken at any time during the experiment. The care and care of the animals is carried out in accordance with state and federal regulations. Each animal is identified by a number placed on the front of the cage and on the rat's tail.

B. Experimental layout

On day 1, we measured the body weight (BW) and hind paw volume of each rat using the f / PW / mercury displacement method, a computer-connected pressure transducer

·· · ·

- 45 applied ^. On day 0, arthritis was induced at an oil dose of 4.4 mg / kg of Mycobacterium butyricum as follows: the rats were anesthetized and aseptically injected with the same amount of Mycobacterium butyricum in a single SC injection into the base of the tail.

Thereafter, leg volumes and body weights are measured on different days, usually twice a week. For the prophylactic / preventive / protocol, the rats are randomly divided into groups of 8-10 rats and treated daily from day 0 until completion of the experiment. For the therapeutic protocol, the rats were randomly assigned to a treatment group of 8-10 rats, based on the hind paw volume at day 10. Dosing of the drug is started on day 10 and daily until the end of the experiment. For both protocols, the animals were placed in deep-litter shoe boxes on day 10 or before.

The solutions added

For compounds not prone to oxidation

The drugs are weighed on a calibrated balance and mixed with distilled water in a volumetric flask. Adjust the pH of the solution to 7.4 with 0.1 N sodium hydroxide solution. The solution is then filtered through a sterile filter of 0.45 µm into a sterile container. When not in use, store the solution in a refrigerator.

For compounds susceptible to oxidation

The drugs are weighed on a calibrated balance and mixed with deoxygenated water in a volumetric flask.

The stock solution was sterile filtered through a 0.45 µm sterile filter. When not in use, dilute the stock solution.

Remove a stock-specific amount of the stock solution in a container cooler daily, place in a small beaker and adjust the pH to 7.4 according to pre-calculated calculations.

The adjusted solution can be further diluted as necessary / with deoxygenated water.

The drugs are calculated based on the molecular weight, the purity of the compound, the amount based on mg / kg body weight and the desired final concentration in mg P / kg. The volume administered was 0.1 ml / 100 g body weight per rat, injected subcutaneously into the groin of the animal, alternating daily; or a dose of 1 ml / 200 g body weight orally using a bent stainless steel dosing tube. We make weekly adjustments based on changes in body weight.

X-rays, necropsy and tissue collection

IRISH)

Finally, all rats were sacrificed by intraperitoneal administration of 1 ml Socomb. Immediate radiography of the whole body is then performed (Torrox 120D X-ray unit, MA = 5, ISOP = 50, time 60 sec, on Kodak non-screen medical film). The hind legs of each rat are removed and

Fixed in a 10% buffered formaldehyde solution together with a piece of liver, kidney, spleen and thimus (thymus). The stomach joints were descaled in 4% EDTA (pH 7.4) and routinely preserved in paraffin blocks and H + E staining. The organ parts are also preserved in paraffin and stained (H + E).

Histological sections were qualitatively evaluated for bone and soft tissue disorders using a light microscope. X-rays for bone resorption were assigned to six anatomical trabecular bone sites on each hind foot and four positions on each front foot on a 0-3 scale, giving a score of 0-60 for each of the four feet. For reactive new bone formation, X-rays were graded on a 0-3 severity scale for the lateral / lateral and medical surfaces of the tibia, and on a 0-2 scale for each of the other sites mentioned above, giving a 0-44 arbitrary score.

D. Statistical Analysis

Data analysis of foot volume, bone resorption, and reactive new bone formation was performed using the Student's t-test and one-way analysis of variance / square mean deviation / £ 7 ^Tu keys / SAS / / 12 / J. The differences are significant at p = 0.05 or less.

This model provides in vivo data on the efficacy of anti-arthritic compounds that reduce leg swelling, bone loss, and reactive new bone formation compared to arthritic animals treated with saline alone.

8i__példa

Capsules containing the following preparation:

mg / capsule

N- (3-Hydroxy-3,3-diphosphonopropyl) -N, N, N-trimethylammonium chloride 350

Binders / Excipients /

Lactose

90.0

Microcrystalline cellulose

60.0

Magnesium stearate

1.0

Capsules containing the above formulation are prepared by the following conventional methods.

The active ingredient is mixed with the microcrystalline cellulose in a turn shell blender for about 10 minutes.

The mixture thus obtained is passed through a hammer mill equipped with a 80 mesh screen.

The mixture is then returned to the mixer along with the lactose and stirred for about 15 minutes. Magnesium stearate is then added to the mixture and the mixture is stirred for another 5 minutes. The mixture thus obtained is then compressed on a piston-operated capsule-filler.

The capsules so prepared contain the active ingredient

1-5. may be substituted by any other compound prepared according to example.

- 49 9 _r __example

Tablets with the following composition:

agent

N- / 4-Hydroxy-4,4-diphosphono-butyl / -N, N-dimethyl-N- / 2-mercapto-ethyl / ammonium chloride

Binders / Excipients /

Lactose / spray dried /

Starch / 1500 /

Maq-Sodium Stearate mg / tablet

700.0

200.0

100.0

25.0

Tablets of the above composition are prepared by the following conventional methods.

The active ingredient is ground in a ball mill for about one minute. The milled active ingredient is then mixed with the spray dried lactose in a twin paddle blender for about 20 minutes. The starch is added to the mixture and stirred for another 15 minutes. The mixture is compressed into tablets using a standard tabletting machine.

The tablets thus prepared contain the active ingredient

1-5. may be substituted by any other compound prepared according to example.

10 _Λ __Example

Preparing injections

Injectable solutions are prepared by conventional procedures, with 10.0 ml of physiological saline and N- / 4.

Using 50-hydroxy-4,4-diphosphonobutyl / -N, N, N-trimethylammonium chloride and adjusting the pH to 7.4.

An injection once daily for 4 days results in a significant improvement in rheumatoid arthritis in patients weighing approximately 70 kg.

In the injectable solutions prepared in this manner, the active ingredient is prepared according to the method of claims 1-5. may be substituted by any of the compounds prepared according to example.

ll _x __example

A 72-year-old Caucasian male of about 92 kg had moderate to severe pain and occasionally had swelling in his right knee. After about 1 year of steadily increasing malaise, he visited a doctor who provided a clinical diagnosis of right knee osteoarthritis, which was confirmed by X-rays.

After treatment with a variety of drugs to improve his condition, such as aspirin, naprosen and ketoprofen, his symptoms worsened again and his condition seemed to degenerate. He went back to the doctor, who then prescribed the Example 9 talbets twice a day, 2 hours before or after meals for 3 months. The clinical symptoms of pain and swelling, notably after prolonged walking, improved significantly after 3 months of treatment. After the three-month dose of two tablets had been completed for three months, treatment was continued with half of the originally prescribed dose, i.e., one capsule as described in Example 8, for an indefinite period of time.

- 51 12 ..__ Example

A 55-year-old 55-year-old black woman had swollen and deformed finger joints in both hands, partially losing the strength and / or dexterity of her fingers and hands. Visual and X-ray examination and various appropriate clinical studies confirmed by the American Rheumatology Association (ARA) have confirmed rheumatoid arthritis in the woman.

After an unsuccessful painkiller and anti-inflammatory therapy, her doctor prescribed her the tablets of Example 9 twice a day, two hours before or after meals for four months. One. after months of treatment, swollen finger joints! her symptoms were significantly improved and her finger movement significantly increased. The woman continued treatment until the end of the fourth month, after which her doctor prescribed the same dose for another two months.

13λ__ £ Eldad

A 12-year-old girl of Spanish origin weighing approximately 37 kg reported to the doctor with primary adolescent rheumatoid arthritis. Symptoms: severe inflammation of the joints, complicated by heat and pressure, and rapid and pathological deterioration of joint function.

She was referred by her doctor to a rheumatologist, who immediately prescribed a vigorous therapy to her, receiving the solution of Example 10 intravenously for three days, with one injection per day for 2 hours. At the end of the intravenous course, the doctor described the tablets prepared as in Example 9 • · «··» ·

- 52 for two months, during which time the patient showed significant improvement, increased mobility and reduced pain. For the next two months, the doctor reduced the dose to 3/4 of the original oral dose, prescribing three tablets for two days, ie two for one day and one for the next. At the end of this course, the doctor again reduced the dose to one-quarter of the original dose of one of the tablets of Example 9 for a further four months.

14 Example _Λ __

A 62-kg, 60-year-old Caucasian woman had severe back pain. Your doctor diagnosed you with an x-ray that your L ^, l vertebra was probably broken by osteoporosis. The patient was prescribed a three-month course of treatment and received a 700 mg tablet of Example 9 once daily. You should take the 700 mg tablet 2 hours before or after a meal. After three months, the dose was reduced to a 350 mg capsule prepared according to Example 8, which was taken every other day for three months. The doctor then took the patient to a maintenance course, taking the 100 mg capsule prepared as in Example 8 daily for six months. After six months of maintenance treatment, the patient no longer experienced back pain and the X-ray showed no fracture.

- 53 »· * ·

15 ^ __ Field

A 75-year-old, 53 kg eastern woman fell and her hip fractured. He was hospitalized and had osteoporosis. As a treatment, she was prescribed injections of calcitonin. Calcitonin injections are painful for the patient and could not adapt to this calcitonin treatment. At this point, your doctor switched your treatment to an oral phosphonate cure. You received a 700 mg tablet of Example 9 twice daily for one month. After one month of treatment, you received one 700 mg tablet once a day for two months. After two months, he took a 100 mg capsule prepared as in Example 8 daily for three months. The medical examination then found that the mineral density of the forearm had not noticeably decreased, as demonstrated by photonabsorption.

16 _r __example

An 85-year-old, 65-kg American native man reported to the doctor with severe back pain. The X-ray revealed the collapse of several smaller vertebral bodies, which resulted from significant bone loss due to osteoporosis. The patient was prescribed a two-month course and on the same day, taken eight hours apart, a 700 mg tablet and a 350 mg

They were prepared according to Example 8. After a two-month course, this dose is reduced to a 350 mg capsule once daily for two months. He was then x-rayed and another fractured

• • · · »« ·· ·· ♦ · ·· ♦ • «· ♦ . · ·. · ·· " ··. · ..

observed. He was then treated for maintenance treatment and received once daily for six months, prepared according to Example 8

100 mg capsules. No significant decrease in bone density was observed after six months.

Claims (10)

Claims CLAIMS 1. Compounds of formula (I) containing a quaternary nitrogen atom and pharmaceutically acceptable salts and esters thereof for use in the treatment or prevention of abnormal calcium and phosphate metabolism, wherein the substituents have the following meanings: m is an integer from 0 to 10; n is an integer from 1 to 10; and m + n = 1-10; a / R is nothing, -SR ^, R ^ SR ^, H, an optionally 'substituted with three C1-8 alkyl, -0R, -CO ₂ R ^3, -o2cr ³ -n / r ^3/2, n / R ³ / C / O / R ³ c / p / n / r _^3/2, 3, halo, C / O / R, nitro, or hydroxy, optionally substituted saturated monocyclic or polycyclic heterocyclic ring or an optionally substituted saturated monocyclic or polycyclic carbocyclic ring, preferably optionally substituted saturated monocyclic or polycyclic heterocyclic ring, optionally substituted saturated monocyclic or polycyclic carbocyclic rings and nil; hydrogen, -SR ^ SR ^ and -R ^9; b / R ^ is -SR ^, -R ^ SR ^, hydrogen, optionally 3-substituted C1-8 alkyl, -0R, -CO ₂ R ^3, -o2cr ³ -n / r ^3/2, -N / R ³ / C / O / R ^3, C / O / N / r _^3/2, 3, halo, C / O / R, nitro, or hydroxy, optionally substituted saturated monocyclic or ···· ·· · • • ♦ ♦ «· ··· polycyclic heterocyclic ring, optionally substituted saturated monocyclic or a polycyclic carbocyclic ring, optionally substituted, unsaturated monocyclic or polycyclic heterocyclic ring, optionally substituted unsaturated monocyclic or polycyclic carbocyclic ring 6 9 6 or combinations thereof, preferably -SR, -R SR, hydrogen, optionally substituted C 1 -C 8 alkyl or optionally substituted saturated monocyclic or polycyclic heterocyclic ring or optionally substituted saturated monocyclic or polycyclic carbocyclic ring; c / each R is optionally substituted C 1 -C 35 alkyl, optionally substituted phenyl 96, benzyl or -R SR, preferably optionally substituted C 1 -C 35 alkyl or -R SR; d / R H, optionally substituted C 1-8 -C 6 alkyl, or -R SR, preferably hydrogen; e / R ^{6 is} hydrogen, -C / O / R ⁷ , -C / S / R ⁷ , -C / O / N / R ⁷ , -C / O / OR ⁷ , 77 77. -C / S / N / R / ₂ , -C / S / OR, wherein R is hydrogen or optionally substituted C 1 -C 8 alkyl; preferably R is hydrogen, -C / O / R or -C / S / R; f / R is -COOH, -SO ₃ H, -PO ₃ H _2, or -P / O // OH / R ⁴ , ***: ········································································································· »· * ··· ·· ♦ * ·· 4- wherein R is C 1-3 alkyl; preferably R is -PO ₃ ^h ₂ or -P / O / OH / R ⁴ ; 9-g / R is optionally substituted C 1 -C 8 alkyl; and 8 to 6 96, h / R hydrogen, halogen, -SR, -R SR, amino, hydroxy or optionally substituted C1-C8 alkyl, preferably hydrogen or -SR. Compounds according to claim 1, wherein R 1 is an optionally substituted saturated monocyclic or polycyclic heterocyclic ring or an optionally substituted saturated monocyclic or polycyclic carbocyclic ring. Compounds according to claim 1 which are R 6 is hydrogen, -SR, -R SR, optionally substituted C 1 -C 8 alkyl, or optionally substituted saturated monocyclic or polycyclic heterocyclic rings. Compounds according to claim 1 which are In the formula 1 6 9 6, R is nothing, -SR, -R SR, or hydrogen. Compounds according to claim 1, wherein R 1 is -SR 4, -R 4 SR 5, hydrogen or optionally substituted C 1 -C 8 alkyl. The compound of claim 1, wherein R ^{5 is} optionally substituted C 1 -C 8 alkyl. • ·· ♦ · · · * ·· • • ·· » · · · • • • 9 · · · • «· • • 9 999 Compounds according to claim 1 which are 5, 9 6, R is -R SR. Compounds according to claim 1, wherein R 4 is an optionally substituted saturated or unsaturated heterocyclic ring. Pharmaceutical compositions for the treatment of conditions associated with abnormal calcium and phosphate metabolism, comprising a safe and effective amount of a phosphonate compound according to claim 1, b / a pharmaceutically acceptable excipient. Use of a compound according to claim 1 for the preparation of a medicament for the treatment or prevention of abnormal calcium and phosphate metabolic disorders in humans or other mammals in need of such treatment, characterized in that a compound according to claim 1 is safe and effective in an amount of said human or other mammal.