AU675224B2 - Quaternary nitrogen-containing phosphonate compounds, for treating abnormal calcium and phosphate metabolism - Google Patents

Description

OPI nATE 30/12/93 AOJP DATE 10/03/94 APPLN. ID 424 3/93 PCT NUMBER PCT/US93/04469 AU9342453 INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (51) International Patent Classification 5 International Publication Number: WO 93/24494 CO7F'/38, A61K 31/66 Al C07 //59 (43) International Publication Date: 9 December 1993 (09.12.93) (21) International Application Number: PCT/US93/04469 (74) Agents: REED, David et al.; The Procter Gamble Company, 5299 Spring Grove Avenue, Cincinnati, OH (22) International Filing Date: 11 May 1993 (11.05.93) 45202 (US).

Priority data: (81) Designated States: AU, BB, BG, BR, CA, CZ, FI, HU, JP, 07/891,355 29 May 1992 (29.05.92) US KP, KR, KZ, LK, MG, MN, MW, NO, NZ, PL, RO, RU, SD, SK, UA, VN, European patent (AT, BE, CH, DE, DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, (71)Applicant: THE PROCTER GAMBLE PHARMA- SE), OAPI patent (BF, BJ, CF, CG, CI, CM, GA, GN, CEUTICALS, INC. [US/US]; Route 320, Woods Corn- ML, MR, NE, SN, TD, TG).

ers, Norwich, NY 13815-0191 (US).

(72) Inventors: EBETINO, Frank, H. 11249 Acrewood Drive, Published Cincinnati, OH 45249 KAAS, Susan, M. R.D. With international search report.

Box 162, Sherburne, NY 13460 FRANCIS, Marion, David 10018 Winlake Drive, Cincinnati, OH 45231 (US).

7 5224 rS7 404 (54)Title: QUATERNARY NITROGEN-CONTAINING PHOSPHONATE COMPOUNDS, MAL CALCIUM AND PHOSPHATE METABOLISM FOR TREATING ABNOR-

R

2 R R I P0 3

H

R

2 C N- C-R e Rn. R Rn (57) Abstract The present invention relates to quaternary nitrogen-containing phosphonate compounds, and the pharmaceutically-acceptable salts and esters thereof having general formula The present invention further relates to pharmaceutical compositions containing a safe and effective amount of a compound of the present invention, and pharmaceutically-acceptable excipients. Finally, the present invention relates to methods for treating or preventing pathological conditions characterized by abnormal calcium and phosphate metabolism such as osteoporosis and arthritis, especially rheumatoid arthritis, and osteoarthritis, in humans or other mammals. This method comprises administering to a human or other mammal in need of such treatment a safe and effective amount of a compound or composition of the present invention.

WO 93/24494 PCT/US93/04469 QUATERNARY NITROGEN-CONTAINING PHOSPHONATE COMPOUNDS, FOR TREATING ABNORMAL CALCIUM AND PHOSPHATE METABOLISM.

BACKGROUND OF INVENTION This invention relates to novel quaternary, nitrogencontaining phosphonate compounds, including bisphosphonates, phosphonoalkylphosphinates, phosphonocarboxylates, and phosphonosulfonates. This invention also relates to pharmaceutical compositions containing these novel compounds as well as to a method of treating or preventing certain metabolic bone disorders characterized by abnormal calcium and phosphate metabolism by utilizing a compound or pharmaceutical composition of the present invention. Specifically, this invention relates to a method of treating or preventing osteoporosis and arthritis, especially rheumatoid arthritis and osteoarthritis by utilizing a compound or pharmaceutical composition of the present invention.

A number of pathological conditions which can afflict warmblooded animals involves abnormal calcium and phosphate metabolism. Such conditions may be divided into two broad categories.

1. Conditions which are characterized by anomalous mobilization of calcium and phosphate leading to general or specific bone loss, such as osteoporosis and Paget's disease, or excessively high calcium and phosphate levels in the fluids of the body, such as hypercalcemia of tumor ori.gin. Such conditions are sometimes referred to herein as pathological hard tissue demineralizations.

WO 93/24494 PCT/US93/04469 -2- 2. Conditions which cause or result from deposition of calcium and phosphate anomalously in the body, such as arthritis, particularly rheumatoid arthritis and osteoarthritis. These conditions are sometimes referred to herein as pathological calcifications.

The first category includes the most common metabolic bone disorder, osteoporosis; osteoporosis is a condition in which bone hard tissue is lost disproportionately to the development of new hard tissue. Osteoporosis can be generally defined as the reduction in the quantity of bone, or the atrophy of skeletal tissue. Marrow and bone spaces become larger, fibrous binding lecreases, and compact bone becomes fragile. Osteoporosis can be subclassified as menopausal, senile, drug-induced (e.g.

adrenocorticoid, as can occur in steroid therapy); disease-induced (arthritic and tumor), etc.; however, the manifestations are essentially the same.

In general, there are two types of osteoporosis: primary and secondary. "Secondary osteoporosis" is the result of a separate disease process or agent. However, approximately 90% of all osteoporosis cases are "primary osteoporosis". Such primary osteoporosis includes postmenopausal osteoporosis, age-associated osteoporosis, disuse osteoporosis (affecting a majority of individuals over the age of 70 to 80), and idiopathic osteoporosis affecting middle-aged and younger men and women.

For soe osteoporotic individuals, the loss of bone tissue is sufficiently great so as to cause mechanical failure of the bone structure. Bone fractures often occur, for example, in the hip and spine of women suffering from postmenopausal osteoporosis. Kyphosis (abnormally increased curvature of the thoracic spine) may also result.

The mechanism of bone loss in osteoporotics is believed to involve an imbalance in the process of "bone remodeling". Bone remodeling occurs throughout life, renewing the skeleton and maintaining the strength of bone. This remodeling involves the erosion and filling of discrete sites on the surface of bones, by an organized group of cells called "basic multicellular units" or WO 93/24494 PC17US93/04469 -3- "BMUs". BMUs primarily consist of "osteoclasts", "osteoblasts", and their cellular precursors. In the remodeling cycle, bone is resorbed at the site of an "activated" BMU by an osteoclast, forming a resorption cavity. This cavity is then filled with bone by an osteoblast.

Normally, in adults, the remodeling cycle results in a small deficit in bone, due to incomplete filling of the resorption cavity. Thus, even in healthy adults, age-related bone loss occurs. However, in osteoporotics, there may be an increase in the number of BMUs that are activated. This increased activation accelerates bone remodeling, resulting in abnormally high bone loss.

Although its etiology is not fully understood, there are many risk factors thought to be associated with osteoporosis.

These include low body weight, low calcium intake, physical inactivity, and estrogen deficiency.

Current osteoporosis treatment consists primarily of calcium and estrogen administration.

The second category, involving conditions manifested by anomalous calcium and phosphate deposition, includes myositis ossificans progressiva, calcinosis universalis, and such afflictions as arthritis (including, for example, rheumatoid arthritis and osteoarthritis), neuritis, bursitis, tendonitis, and conditions which predispose involved tissue to deposition of calcium.

In addition to osteoporosis, bone loss can result from rheumatoid arthritis and osteoarthritis. Rheumatoid arthritis is a chronic, systemic and articular inflammatory disorder characterized by weakening of the joint capsules and ligaments, followed by destruction of cartilage, ligaments, tendon and bone, and a decrease in viscosity and other alterations in the synovial fluid. Rheumatoid arthritis symptoms include systemic weakness, fatigue, localized pain, stiffness and weakness and swelling and deformation of the joints of the body. Rheumatoid arthritis is most common in women in the fourth to sixth decade of life.

WO 93/24494 PCrI/US93/04469 -4- The pathogenesis of rheumatoid arthritis, leading to the destruction of the joints, is characterized by two phases: 1) an exudative phase involving the microcirculation and the synovial cells that allow an influx of plasma proteins and cellular elements into the joint and 2) a chronic inflammatory phase occurring in the sub-synovium and sub-chondral bone, characterized by pannus (granulation tissue) formation in the joint space, bone erosion, and cartilage destruction. The pannus may form adhesions and scar tissue which causes the joint deformities characteristic of rheumatoid arthritis.

The etiology of rheumatoid arthritis remains obscure.

Infectious agents such as bacteria and viruses have been implicated. A current hypothesis is that the Epstein-Barr (EBV) virus is a causative agent for rheumatoid arthritis.

Current rheumatoid 'arthritis treatment consists predominantly of symptomatic relief by administration of non-steroidal anti-inflammatory drugs. Non-steroidal anti-inflammatory drug treatment is mainly effective in the early stages of rheumatoid arthritis; it is unlikely it will produce suppression of joint inflammation if the disease is present for more than one year., Gold, methotrexate, immunosuppressants and corticosteroids have been tried with limited success.

On the other hand, osteoarthritis, is an inherently non-inflammatory disorder of the movable joints characterized by deterioration and abrasion of articular cartilage, as well as by formation of new bone at the joint surface. As osteoarthritis progresses, the surface of the articular cartilage is disrupted and wear particles gain access to the synovial fluid which in turn stimulates phagocytosis by macrophage cells. Thus, an inflammatory response is eventually induced in osteoarthritis.

Common clinical symptoms of osteoarthritis include cartilaginous and bony enlargements of the finger joints and stiffness on awakening and painful movement.

Common symptomatic treatments for osteoarthritis include analgesics, anti-inflammatories, steroids, and physical therapy.

WO 93/24494 P~7/US93/044Q69 A variety of phosphonic acid derivatives have been proposed for use in the treatment and prophylaxis of diseases involving abnormal calcium and phosphate metabolism. For example, numerous references, all incorporated by reference herein, disclose compositions containing polyphosphonates, in particular diphosphonates such as ethane-l-hydroxy-1,1-diphosphonic acid and their use in inhibiting anomalous deposition and mobilization of calcium and phosphate in animal tissue: U.S.

Patent 3,683,080, issued August 8, 1972 and U.S. Patent 4,230,700, issued October 28, 1980, both to Francis, and U.S.

Patent 4,868,164 to Ebetino, issued September 19, 1989. Numerous other references describe heterocyclic substituted diphosphonic acids useful for the treatment of osteoporosis and/or arthritis, and are hereby incorporated by reference herein: U.S. Patent 5,071,840, to Ebetino, et al., issued December 10, 1991; U.S.

Patent 4,868,164, to Ebetino, et al., issued September 19, 1989; U.S. Patent 5,104,863, to Benedict, et al., issued April 14, 1992; U.S. Patent 4,267,108, to Blum et al., issued May 12, 1981; U.S. Patent 4,746,654 to Breliere et al., issued May 24, 1988; U.S. Patent 4,876,247 to Barbier, et al., issued October 24, 1989, and European Patent Application Publication No.

100,718, of Breliere, published February 15, 1984; European Patent Application Publication No. 170,228, of Boehringer Mannhein GmbH, published February 5, 1986; European Patent Application Publication No. 186,405, of Benedict and Perkins, published July 2, 1986; European Patent Application io. 298,553, of Ebetino, published January 11, 1989; U.S. 4,754,993, to Bosies, et al., issued November 15, 1988; U.S. 4,939,130, to Jaeggi, et al., issued July 3, 1990; U.S. 4,971,958 to Bosies, et al., issued November 20, 1990; WO 90/12017, Dunn, et al.

published October 18, 1990; WO 91/10646, Youssefyeh, et al.

published July 25, 1991; AU-A-26738/88, Jaeggi, publication date June 15, 1989; AU-A-45467/89 of Ciba-Geigy, publication date May 31, 1990.

WO 93/24494 PCF/US93/0~469 -6- Finally, U.S. 4,208,401 to Bauman, issued June 17, 1980, discloses non-heterocyclic ring substituted quaternary ammonium bisphosphonates useful as anti-calculus agents.

DE 40 11 777 to Jaeggi, disclosed October 18, 1990; (DE '777) discloses a heterocyclic ring substituted diphosphonate wherein said heterocyclic ring can be lower alkyl substituted.

Said heterocyclic ring is bridged to the phosphonic acid group via a quaternary non-ring nitrogen atom. DE '777 also discloses that the compounds produce pronounced inhibition of bone resorption and thus are useful in treating osteoporosis, inflammatory and degenerative joint diseases, peridontitis, and hyperparathyroidism. The disclosures of these references are incorporated by reference herein.

The compounds of the present invention have osteoprotective activity at the site of joint destruction in arthritis conditions and have that activity as an additional benefit in the treatment of arthritis over the above merely relieving the symptoms of inflammation. The term "osteoprotecive activity" as used herein means disease-modifying activity on bone and surrounding soft tissue at the site of joint destruction.

It has been surprisingly discovered that the compounds of the present invention, wherein the nitrogen is quaternized, have more potent bone antiresorptive activity and therapeutic utility in treating osteoporosis and rheumatoid arthritis and osteoarthritis than nitrogen-containing compounds where the nitrogen atom is not quaternized. Moreover, the compounds of the present invention exhibit unusual solubility properties. Thus, the compounds of the present invention may be more readily orally absorbed compounds. The more readily absorbed a compound, the more effective it may be at lower doses. Lower doses are generally preferable because undesirable side effects are decreased.

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

WO 93/24494 IPCTUS93/4469 -7- It is a further object of the present invention to provide pharmaceutical compositions useful for the treatment and prophylaxis of osteoporosis and arthritis, especially rheumatoid arthritis and osteoarthritis. In addition, it is an object of the present invention to provide methods for treating or preventing osteoporosis and arthritis, especially rheumatoid arthritis and osteoarthritis.

These and other objects of the present invention will become apparent from the detailed disclosure of the present invention provided hereinafter.

SUMMARY OF THE INVENTION The present invention relates to quaternary nitrogencontaining phosphonate compounds, and the pharmaceutically-acceptable salts and esters thereof having the following general formula: R

R

2

R

s I I

PO

3 H2

R

2 C N- C C-R 8 Rm R wherein m is an integer from 0-10; and n is an integer from 1-10; m n is from 1-10; R1 is-selected from the group consisting of nil; -SR 6

-R

9

SR

6 hydrogen; substituted or unsubstituted CI-C8 alkyl;

-OR

3 -C02R 3 -gtR -NR 3 2; -N(R 3 )C(0)R 3 -C(0)N(R 3 )2; halogen; -C(0)R 3 nitro; hydroxy; substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic rings;

R

5 is selected from the group consisting of -SR 6

-R

9

SR

6 hydrogen; substituted or unsubstituted CI-C8 alkyl; WO 93/24494 PGF/US93/04469 -8-

-OR

3 -C02R 3 -B -NR 3 2; -N(R 3

)C(O)R

3

-C(O)N(R

3 2 halogen; -C(O)R 3 nitro; hydroxy; substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyclic carbocyclic rings and combinations thereof; each R 2 is selected from the group consisting of substituted or unsubstituted C-C35 alkyl; unti-e4t- utcd or- -substituted pheny-l; benzyl; or R 9

SR

6

R

3 is selected from the group consisting of H; unsubstituted or substituted CI-C8 alkyl; R 9

SR

6

R

6 is selected from the group consisting of -H; -C(0)R7; -C(S)R7; -C(O)N(R7) 2 -C(0)OR 7

-C(S)N(R

7 )2;

-C(S)OR

7 where R 7 is hydrogen or unsubstituted or substituted CI-C8 alkyl; R is selected from the group consisting of -COOH; -SO3H; -P03H2; and -P(O)(OH)R 4 where R 4 is an alkyl group having 1-3 carbons.

R

9 is substituted or unsubstituted C1-C8 alkyl;

R

8 is selected from the group consisting of hydrogen, halogen; SR 6

R

9

SR

6 amino; hydroxy; substituted and unsubstituted C 1

-C

8 alkyl; In this general structure, the quaternary nitrogen atom must be linked to the phosphonic acid containing carbon atom via a linking chain. It cannot be bonded directly to the phosphonic acid containing carbon atom.

The present invention further relates to pharmaceutical compositions containing a safe and effective amount of a compound of the present invention, and pharmaceutically-acceptable excipients. Finally, the present invention relates to methods for treating or preventing pathological conditions characterized by ,abnormal calcium and phosphate metabolism such as osteoporosis and arthritis, especially rheumatoid arthritis, and 05/12 '96 THU 16:08 FAX 61 3 9614 1867PILPSOCJD AP-OM STNR (1o PHILLIPS ORMONDE A AIPO-COMMISS'NER 2003 osteoarthritis, in humans or other mammals. This method comprises administering to a human or other mammal In need of such treatment a safe and effective amount of a copmound or composition of the present Invention.

In a further aspect the invention provides a pharmaceultical composition comprising: a safe and effective amount of a quaternary nitrogen-containing phosphonate compound having the general formula M 2 wherein m is an integer from 0-10; and n is an integer from 1-10; m ±n is from 1-10; Riis select~pd from the group consisting of nil -SR 6 6 -R SR hydrogen; substituted orunsub~stituted C 1 -C alkyl; -O -CO R ;-NR 2 -N(R _C(O)N(R 3 *:halogen; nitro; hydroxy; substituted or unsubstituted saturated inonocyclic or polycyclic ****hoterof.cyclic rings; substitute.1 or unsubstituted saturated monocyclic or polycyclic carbocyclic rings-, R5is selected from the group consisting of -R

-R

9

SR

6 hydrogen; substituted or unsubstituted C 1

-C

8 alkyl;

-OR

3 -CO 2 R 3; -NR 3 2 halogen; nitro,, hydroxy; substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic rings; substituted or un -,substituted saturated rnonocyclic or polycyclic carbocyclic rings; substituted or unsubstituted unsaturated monocyciic or polycyclic heterocyclic rings; substituted or unsubstituted unsaturated nionocyclic or poiycyclic carbocyClic rings and combinations thereof; '1111' Th 10.-08 FAX 01 1001.4 1407 III1111[1AP OM')NOU I APO M 9. NV C 01 0 0 4 ()each R is su'1ectad from the group consisting of substituted or unsubstituted C 1 C: S alkyl; benzyl; or R 9

SR

5

R

3 is selecttd from the group consisting of H; 6nusiue orsutituted rC 1

-C

8 alkyl; R 9 SR 6

R

5 is selected from the group consisting of -H; where R7 is ,hydrogen or unsubstituted or substituted C I'C, alkylo; R is selected from the group consisting of -P0 3

H

2 and -P(O)(OH)R 4 whe-"R is an alkyl group having 1-3 carbons.

R9 is substituted or unsubstituted C alkyl; R 8 is selectead from the grotip consisting of hydrogen, :::halogen; SF1 6

R

9 S 6 amino, hydroxy; substituted and unsubstituted C-C~ alkyl; and a pharmaceutir:Aily-acceptable excipient.

WO 93/24494 1'CI'7 US93/04469 -9comprises administering to a human or ii--ina n need of such treatment effective amount of a compound or Definitions and Usage of Terms The following is a list of definitions for terms used herein.

"Heteroatom" is a nitrogen, sulfur, or oxygen atom. Groups containing one or more heteroatoms may contain different heteroatoms.

"Alkyl" is an unsubstituted or substituted, straight-chain or branched, saturated or unsaturated hydrocarbon chain, said hydrocarbon chain may be saturated having 1 to 8 carbon atoms, and preferably, unless otherwise stated, from 1 to 4 carbon atoms; said hydrocarbon chain may be unsaturated, having 2 to 8 carbon atoms, and preferably, unless otherwise stated, 2 to 4 carbon atoms. Accordingly, the term "alkyl", as used herein, encompasses alkenyl hydrocarbon unsaturated chains having at least one olefinic double bond and alkynyl hydrocarbon unsaturated chains having at least one triple bond. Preferred alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, and butyl.

"Carbocyclic ring" or "Carbocycle" as used herein is an unsubstituted or substituted, saturated, unsaturated or aromatic, hydrocarbon ring; Carbocyclic rings may be monocyclic or polycyclic: Monocyclic ring generally contain from 3 to 8 atoms, preferably 5 to 7 atoms. Polycyclic rings containing two rings contain 6 to 16, preferably 10 to 12, atoms and those with three rings generally contain 13 to 17, preferably 14 to 15, atoms.

"Heteroalkyl" is an unsubstituted or substituted, saturated chain having from 3 to 8-members and comprising carbon atoms and one or two heteroatoms.

"Heterocyclic ring" or "Heterocycle" as used herein is an unsubsti.tuted or substituted, saturated, unsaturated or aromatic WO 93/24494 PCO/WS93/04469 ring comprised of carbon atoms and one or more heteroatoms in the ring. Heterocyclic rings may be monocyclic or polycyclic rings.

Monocyclic rings generally contain from 3 to 8 atoms, preferably 5 to 7 atoms. Polycyclic ring systems consisting of two rings generally contain 6 to 16, preferably from 10 to 12 atoms. Polycyclic ring systems consisting of three rings generally contain 13 to 17 atoms, preferably 14 to 15 atoms. A heterocyclic ring moiety may consist of heterocycles or heterocycles and carbocycles. Each heterocyclic ring moiety must have at least one nitrogen atom. Unless otherwise stated any additional heteroatoms may be independently chosen from nitrogen, sulfur, and oxygen.

"Aryl" is an aromatic carbocyclic ring. Preferred aryl groups include, but are not limited to, phenyl, tolyl, xylyl, cumenyl, and naphthyl.

"Heteroaryl" is an aromatic heterocyclic ring. Preferred heteroaryl groups include, but are not limited to, thienyl, furyl, pyrrolyl, pyridinyl, pyrazinyl, oxazolyl, thiazolyl, quinolinyl, pyrimidinyl, and tetrazolyl.

"Alkoxy" is an oxygen atom having a hydrocarbon chain substituent, where the hydrocarbon chain is an alkyl or alkenyl -0-alkyl or -0-alkenyl). Preferred alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and alkyloxy.

"Hydroxyalkyl" is a substituted hydrocarbon chain which has a hydroxy substituent and may have other substituents. Preferred hydroxyalkyl groups include, but are not limited to, hydroxyethyl, hydroxypropyl.

"Carboxyalkyl" is a substituted hydrocarbon chain which has a carboxy substituent -COOH) and may have other substituents. Preferred carboxyalkyl groups include carboxymethyl, carboxyethyl, and their acids and esters.

"Aminoalkyl" is a hydrocarbon chain alkyl) substituted with an amine moiety NH-alkyl-) such as aminomethyl.

"Alkylamino" is an amino moiety having one or two alkyl substituents -N-alkyl) such as dimethylamino.

WO 93/24494 IC/U9/46 "Al kenyl amino" is an amino moiety having one or two alkenyl substituents -N-alkenyl).

"Al kynal amino" is an amino moiety having one or two alkynyl substituents -N-alkynyl).

"Alkylimino" is an imino moiety having one or two alkyl substituents -N-alkyl-).

"Arylalkyl" is an alkyl moiety substituted with an aryl group. Preferred arylalkyl groups include benzyl and phenyl ethyl.

"Arylamino" is an amine moiety substituted with an aryl group -NH-aryl).

"Arylaxy" is an oxygen atom having an aryl substituent -0-aryl).

"Acyl" or "carbonyl" is a carbon to oxygen double bond, e.g.

Preferred acyl groups include, but are not limited to, acetyl, propionyl, butanoyl, and benzoyl.

"Acyloxy" is an oxygen atom having an acyl substituent -0-acyl); for example, -0-C(-O)-alkyl.

"Acylamino" is an amino moiety having an acyl substituent -N-acyl); for example, -NH-(C-O)-alkyl.

"Halo", "halogen", or "halide" is a chloro, bromo, fluoro, or iodo, atom radical. Chioro, bromo, and fluoro are preferred hal ides.

As referred to herein, a "lower" hydrocarbon moiety "lower" alkyl) is a hydrocarbon chain comprised of from, unless otherwise stated, 1 to 6, preferably from 1 to 4, carbon atoms.

Also, as used herein, the term "thio-substituent" (SR 6 or

R

9

SR

6 includes thiols where R 6 thioesters [-SC(0)R 7 where R 6 uC(O)R 7 dithioesters [-SC(S)R7] where R 6 -C(S)R7; thiocarbamates [-SC(0)N(R 7 where R 6 -C(0)N(R 7 )2; dithiocarbamates [-SC(S)N(R 7 where R 6

-C(S)N(R

7 )2; thiocarbonates [-SC(0)OR 7 where R 6 -C(0)OR7; and dithiocarbonates [-SC(S)OR7] where R 6 uC(S)0R 7

R

7 is generally a hydrogen or Cj-C 8 alkyl. Any of the SR 6 substituents may themselves be substituted with an R9 moiety, i.e. R 9

SR

6 where R 2 is a substituted or unsubstituted Cj-CS alkyl. Accerdingly, WO 93/24494 PCT/US93/04469 -12additional thio-substituents denoted by R 9

SR

6 are alkylthiols, alkylthioesters, alkyldithioesters, alkylthiocarbamates, alkyldithiocarbmates, alkylthiocarbonates, and alkyldithiocarbonates.

The terms "bisphosphonate" or "bisphosphonic acid" as used herein relate to those phosphonate or phosphonic acid compounds that have two phosphonate groups attached to the same carbon atom and are used interchangeably with the terms "diphosphonate" and "diphosphonic acids." Using the structures described herein, the moiety R is PO3H2.

As used herein, the term "phosphonic acid carbon" refers to the carbon atom to which a phosphonic acid group (P03H2) is attached. When another phosphonic acid group is attached to said carbon atom, the resulting compound is bisphosphonate. When a sulfonate group is attached to said carbon atom, the resulting compound is a phosphonosulfonate. When a carboxylate group is attached to said carbon atom, the resulting compound is a phosphonocarboxylate. When a phosphinic acid group is attached to said carbon atom, the resulting compound is a phosphonoalkylphosphinate.

A "pharmaceutically-acceptable" salt is a catonic salt formed at any acidic carboxyl) group, or an anionic salt formed at any basic amino) group. Many such salts are known in the art, as described in World Patent Publication 87/05297, Johnston et al., published September 11, 1987, hereby incorporated by reference herein. Preferred catonic salts include the alkali-metal salts (such as sodium and potassium), and alkaline earth metal salts (such as magnesium and calcium).

Preferred anionic salts include the halide (such as chloride), acetate and phosphate salts.

A "biohydrolyzable ester" is an ester of the quaternary nitrogen-containing heterocyclic phosphonate compounds that does not interfere with the therapeutic activity of the compounds, or that is readily metabolized by a human or other mammal. Many such esters are known in the art, as described in World Patent Publication 87/05297, Johnston et al., published WO 93/24494 PC'/JS93/0"469 -13- September 11, 1987, and hereby incorporated by reference herein.

Such esters include lower alkyl esters, lower acyloxyalkyl esters (such as acetoxymethyl, acetoxyethyl, aminocarbonyloxymethyl, pivaloyloxymethyl, and pivaloyloxyethyl esters), lactonyl esters (such as phthalidyl and thiophthalidyl esters), lower alkoxyacyloxyalkyl esters (such as methoxycarbonyloxymethyl, ethoxycarbonyloxyethyl and isopropoxycarbonyloxyethyl esters), alkoxyalkyl esters, choline esters, and acylamino alkyl esters (such as acetamidomethyl esters).

As defined above and as used herein, substituent groups may themselves be substituted. Such substitution may be with one or more substituents. Such substituents include, but are not limited to, those listed in C. Hansch and A. Leo, Substituent Constants for Correlation Analysis in Chemistry and Biology (1979), hereby incorporated by reference herein. Preferred substituents include, but are not limited to, alkyl, alkenyl, alkoxy, hydroxy, oxo, amino, aminoalkyl aminomethyl, etc.), cyano, halo, carboxy, alkoxyacetyl carboethoxy, etc.), thio, thiol, aryl, cycloalkyl, heteroaryl, heterocycloalkyl piperidinyl, .orpholinyl, piperazinyl, pyrrolidinyl, etc.), imino, thioxo, hydroxyalkyl, aryloxy, arylalkyl, and combinations thereof.

DETAILED DESCRIPTION OF THE INVENTION Novel Quaternary Nitrogen-Containing Phosphonate Compounds The compounds of the present invention are quaternary nitrogen-containing phosphonate compounds, and the pharmaceutically-acceptable salts and esters thereof, having a quaternary nitrogen atom. The quaternary nitrogen atom is bonded to the phosphonic acid containing carbon via a linking chain to the phosphonic acid containing carbon.

The carbon atom which has the phosphonic acid group attached to it may be unsubstituted a hydrogen atom) or substituted. The phosphonic acid carbon may contain two phosphonate groups, rendering a bisphosphonate compound; a phosphonate group WO 93/24494 PCr/US93/044699 -14and an carboxylate group, rendering a phosphonocarboxylate compound; a phosphonate group and a sulfonate group, rendering a phosphonosulfonate compound, a phosphinate group and a phosphonate group, rendering a phosphonoalkylphosphinate compound.

Thus, the quaternary nitrogen-containing phosphonate compounds of the present invention, and the pharmaceutically-acceptable salts and esters thereof, have the general structure: R1 R 2

R

P03H2 I I I /POH

R

2 c N C C-R 8 1 I R R'm R 2

R

5 n In this general structure, R 1 is selected from a variety of non-ring moieties such as nil, -SR 6

-R

9

SR

6 hydrogen, alkyl having 1-8 carbons, -OR 3 -C02R 3 -t21, -NR 3 2, -N(R 3

)C(O)R

3 -C(0)N(R 3 halogen, -C(0)R 3 nitro, hydroxy, substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic rings, substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic rings.

Also, in this general structure, the quaternary nitrogen atom is linked to the phosphonic acid carbon by a linking chain.

Further, in this general structure, n, which is an integer from 1-10, represents said linking chain.

Said linking chain members are selected from a variety of R moieties. R 5 can be -SR 6

-R

9

SR

6 hydrogen, C1-CB alkyl, -OR 3 -C02R 3 4it0 -NR 3 2; -N(R 3

)C(O)R

3

-C(O)N(R

3 halogen, -C(0)R 3 nitro; hydrogen; unsubstituted or substituted saturated monocyclic or polycyclic heterocyclic ring, unsubstituted or substituted saturated monocyclic or polycyclic carbocyclic rings, unsubstituted or substituted unsaturated monocyclic or polycyclic VO 93/24494 VC 9324494PCr/US93/04469 heterocyclic rings, unsubstituted or substituted unsaturated monocyclic, or. polycyclic carbocyclic, rings and combinations thereof.

Finally, in the quaternary nitrogen containing phosphonate compounds of the present invention, R can be -COOH, -SO3H, -P03H2 and -P(O)(OH)R 4 where R 4 is C 1

-C

8 alkyl. Preferred R is P03H2 and P(O)(OH)R 4

R

8 is a substituent on the phosphonic acid containing carbon selected from hydrogen, halogen, SR 6

R

9

,SR

6 amino, hydroxy, substituted and unsubstituted Cj-C8 alkyl.

Preferred R 8 is hydroxy, halogen and amino. R 2 is substituted or unsubstituted Cl-C 3 5 alkyl, substituted or unsubstituted phenyl, benzyl; or R 9

SR

6 Preferred R 2 is substituted or unsubstituted C-CB alkyl and

R

9

SR

6 Preferred quaternary nitrogen-containing phosphonates having an R 1 moiety selected from the RIl moieties described herein before include,

CH

3 I+ PO

H

2 3

(CH

2 3

C

2 _N0+ r

CH

3 P0 3

H

2 N-(3-hydroxy-3,3-diphosphonopropyl)-N,N-dimethyl-N-pentylammonium chloride

CH

3 +1 P0 3

H

2

CH

3 OH CF' I P0 3

H

2

CH

3 N-(4-hydroxy-4,4-diphosphonobutyl)-N,N,N-trialkylamm~fonium salt WO 93/24494 WO 9324494PCT/1J593/04469 -16-

OH

3 1+ P0 3

H

2 CH3-N *".LOH

CI*

CH

3 P0 3

H

2 N-(3-hiydroxy-3,3-diphosphonopropyl )-N,N,N-trialkylammonium salts Also, in this general structure, the RI moiety can also be saturated monocyclic or polycyclic heterocycle.

Preferred quaternary nitrogen-containing phosphonates having a satu.,ated monocyclic or polycyclic heterocycle as an R 1 moiety wherein the quaternary nitrogen atom is linked via a linking chain to the phosphonic acid carbon include: H H P(O)(O-I) 2 I~ P(O)(OH) 2 N-(3-hydroxy-3,3-diphosphonopropyl)-N,N-dimethyl-N-(2-piperidinemethyl) ammnonium chloride Additionally, preferred compounds of the present invention include those compounds having the following structures:

R

2 R2_ I+ P0 3

H

2

R

2 P0 3

H

2 WO 93/24494 WO 9324494PCT/US93/04 469 -17-

R

2 R+ I P0 3

H

2

O'

2

-RN

P0 3

H

2 Preferred compounds of the present invention also include the thia-substituted quaternary nitrogen containing phosphonates.

R

2 R SR 9 N P0 3

H

2

R

2 P0 3

H

2

R

2 9R R2 I+ P03 NtL.j

.H

R2 P0 3

H

2 R 2 +1 P0 3

H

2

R

2 P0 3

H

2

R

2

R

9

SR

6 +1 ~{P 3

H

2

R

2 -N R 8 12 P0 3

H

2 Specific examples of compounds of the present invention i ncl ude: N-(4-hydroxy-4,4-diphosphonobutyl)-N,N,N-trimethyl ammoniumn iodide; N-(3-hydroxy-3,3-diphosphonopropyl)-N,N-dinethyl-N-pentyl ammnonium iodide; 35N-(3-hydroxy-3,3-diphosphonopropyl)-.N,N,N-trimethyl ammronium Siodide;' N-cycloheptyl-N,N-diniethyl-N-(diphosphonomethyl) ammnonium iodide; PCr/US93/04469 WO 93/24494 -18- N-(2-acetylthioethyl)-N-(4-hydroxy-4,4-diphosphonobutyl)-NN-dimethyl ammunkum bromide; N-(2-acetylthioethyl)-N-(3-hydroxy-3,3-diphosphonopropyl)-N-methyl-N-pentyl ammonium bromide; N-(4-hydroxy-4,4-diphosphonobutyl)-N-(3-mercaptopropyl)-NN-dimethyl ammonium chloride; N-(4-hydroxy-4,4-diphasphonobutyl)-N-(mercaptomethyl)-NN-dimethyl ammonium chloride; N-(4-hydroxy-4,4-diphosphonobutyl)-N-(4-methoxybutyl)-NN-dimethyl ammonium chloride; N-(4-hydroxy-2-mercapto-4,4-diphosphonobutyl)-NNN-trimethyl amnonium chloride; N-(4-hydroxy-2-acetylthio-4,4-diphosphonobutyl)-NNN-trimethyl ammonium chloride; N-(3-hydroxy-2-mercapto-3,3-diphosphonopropyl)-N,N-diine.thyl-Npentyl anmonium chloride; N-(3-hydroxy-2-acetylthio-3,3-diphosphonopropyl)-NN-dimethyl-Npentyl ammonium chloride; N-(3-hydroxy-3,3-diphosphonopropyl)-N-methyl-N-pentyl-N-(2-(3pyridyl)ethyl) amnonium chloride; N-cycloheptyl-N-(2-mercaptoethyl)-N-methyl-N-(diphosphonomethyl) ammonium chloride; N-cycloheptyl-N-(mercaptomethyl)-N-methyl-N-(dlphosphonomethyl) ammonium chloride; N,N-dlmethyl-N-(4,4-diphosphonobutyl)-N-(2-(3-piperidinyl)ethyl) ammonium chloride; In order to determine and assess pharmacological activity, testing of the phosphonate compounds in animals is carried out using various assays known to those skilled in the art. Thus, the In vivo bone antiresorptive activity may be conveniently demonstrated using an assay designed to test the ability of these compounds to inhibit the resorption of bone, which bone resorption is characteristic of abnormal calcium and phosphate metabolism. One such test known to the art is the Schenk model.

Another -useful art-known test is the adjuvant arthritis test.

Also useful is the in vitro hydroxyapatite crystal growth VO 93/24494 PCT/US93/0469 -19inhibition test. These and other appropriate tests for pharmacological activity are disclosed and/or referred to in Shinoda et al., Calcified Tissue International, 35, pp 87-99 (1983); Schenk et al. Calcified Tissue Research, 11, pp 196-214 (1973); Russell et al., Calcified Tissue Research, 6, pp 183-196 (1970); Muhlbauer and Fleisch, Mineral Electrolyte Metab. 5 pp 296-303 (1981); Nancollas et al., Oral Biol., 15, 731 (1970); U.S. Patent 3,683,080, to Francis, issued August 8, 1972; U. S.

Patent 4,134,969, to Schmidt-Dunker, Issued January 16, 1979; and EPO Patent Application Publication No. 1891662, published August 6, 1986; the disclosures of all these articles and patent specifications being incorporated herein by reference in their entirety. Certain of these tests for pharmacological activity are also described in more detail in the Examples provided hereinafter.

In addition to being useful for treating or preventing pathological conditions characterized by abnormal calcium or phosphate metabolism, the compounds of the present invention may have other uses. For example, the compounds of the present invention are believed to be useful as bone scanning agents after labeling with 99m-technetium. In addition, the compounds of the present invention are useful as sequestering agents for polyvalent metal ions, particularly di-(e.g. calcium and magnesium) and trivalent indium) metal ions. Thus, the compounds of the present invention are useful as builders in detergents and cleansers, or for treating water. They are also useful as stabilizers for compounds. In addition, they may be useful in preventing the formation of tartar calculus) and/or plaque on teeth. Finally, the compounds of the present invention may be useful as herbicides which are non-toxic to animals.

The quaternary nitrogen-containing phosphonates to be included in the pharmaceutical compositions of the present invention can be made according to the following non-limiting Examples I1 to WO 93/24494 I-IC/US93/04469 Comoositions Containing Novel Quaternary Nitrogen Containing Phosphonate Compounds The novel quaternary nitrogen-containing phosphonate compounds of the present invention may be administered to humans or other mammals by a variety of routes, including, but not limited to, oral dosage forms and injections (intravenous, intramuscular, intraperitoneal and subcutaneous). Numerous other dosage forms containing the novel quaternary nitrogen-containing phosphonate compounds of the present invention can be readily formulated by one skilled in the art, utilizing the suitable pharmaceutical excipients as defined below. For considerations of patient compliance, oral dosage forms are generally most preferred.

The term "pharmaceutical composition" as used herein means a combination comprised of a safe and effective amount of the quaternary nitrogen-containiin. phosphonate compound active ingredient, or mixtures thereof, and pharmaceutically-acceptable excipients.

The phrase "safe and effective amount", as used herein, means an amount of a compound or composition large enough to significantly positively modify the symptoms and/or condition to be treated, but small enough to avoid serious side effects (at a reasonable benefit/risk ratio), within the scope of sound medical judgment. The safe and effective amount of active ingredient for use in the pharmaceutical compositions to be used in the method of the invention herein will vary with the particular condition being treated, the age and physical condition of the patient being treated, the severity of the condition, the duration of the treatment, the nature of concurrent therapy, the particular active ingredient being employed, the particular pharmaceutically-acceptable excipients utilized, and like factors within the knowledge and expertise of the attending physician.

The term "pharmaceutically-acceptable excipients" as used herein includes any physiologically inert, pharmacologically inactive material known to one skilled in the art, which is compatible with the physical and chemical characteristics of the WO 93/24494 PCFUS93/04469 -21particular quaternary nitrogen-containing phosphonate compound active ingredient selected for use. Pharmaceutically-acceptable excipients include, but are not limited to, polymers, resins, plasticizers, fillers, binders, lubricants, glidants, dibintegrants, solvents, co-solvents, buffer systems, surfactants, preservatives, sweetening agents, flavoring agents, pharmaceutical grade dyes or pigments, and viscosity agents.

The term "oral dosage form" as used herein means any pharmaceutical composition intended to be systemically administered to an individual by delivering said composition to the gastrointestinal tract of an individual, via the mouth of said individual. For purposes of the present invention, the delivered form can be in the form of a tablet, coated or non-coated; solution; suspension; or a capsule, coated or non-coated.

The term "injection" as used herein means any pharmaceutical composition intended to be systemically administered to a human or other mammal, via delivery of a solution or emulsion containing the active ingredient, by puncturing the skin of said individual, in order to deliver said solution or emulsion to the circulatory system of the individual either by intravenous, intramuscular, intraperitoneal or subcutaneous injection.

The rate of systemic delivery can be satisfactorily controlled by one skilled in the art, by manipulating any one or more of the following: the active ingredient proper; the pharmaceutically-acceptable excipients; so long as the variants do not interfere in the activity of the particular active ingredient selected; the type of the excipient, and the concomitant desirable thickness and permeability (swelling properties) of said excipients; the time-dependent conditions of the excipient itself and/or within the excipients; the particle size of the granulated active ingredient; 0 93/24494 PCT/US93/04469 -22the pH-dependent conditions of the excipients.

In particular, the solubility, acidity, and susceptibility to hydrolysis of the different quaternary non-ring nitrogen-containing phosphonate active ingredients, such as acid addition salts, salts formed with the carboxylic group, e.g., alkali metal salts, alkaline earth metal salts, etc., and esters, alkyl, aryl, aralkyl, may be used as guidelines for the proper choice. In addition, suitable pH-conditions might be established within the oral dosage forms by adding a suitable buffer to the active ingredient in accordance with the desired release pattern.

As stated hereinabove, pharmaceutically-acceptable excipients include, but are not limited to, resins, fillers, binders, lubricants, solvents, glidants, disintegrants cosolvents, surfactants, preservatives, sweetener agents, flavoring agents, buffer systems, pharmaceutical-grade dyes or pigments, and viscosity agents.

The preferred solvent is water.

Flavoring agents among those useful herein include those described in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, 1990, pp. 1288-1300, incorporated by reference herein. The pharmaceutical compositions suitable for use herein generally contain from 0-2% flavoring agents.

Dyes or pigments among those useful herein include those described in Handbook of Pharmaceutical ExciDients, pp. 81-90, 1986 by the American Pharmaceutical Association the Pharmaceutical Society of Great Britain, incorporated by reference herein. The pharmaceutical compositions herein generally contain from 0-2% dyes or pigments.

Preferred co-solvents include, but are not limited to, ethanol, glycerin, propylene glycol, polyethylene glycols. The pharmaceutical compositions of the present invention include from 0-50% co-solvents.

Preferred buffer systems include, but are not limited to, acetic, .boric, carbonic, phosphoric, succinic, malaic, tartaric, citric, acetic, benzoic, lactic, glyceric, gluconic, glutaric and WO 93/24494 rPCT/US93/04469 -23glutamic acids and their sodium, potassium and ammonium salts.

Particularly preferred are phosphoric, tartaric, citric, and acetic acids and salts. The pharmaceutical composition of the present invention generally contain from 0-5% buffer systems.

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

Preferred preservatives include, but are not limited to, phenol, alkyl esters of parahydroxybenzoic acid, o-phenylphenol benzoic acid and the salts thereof, boric acid and the salts thereof, sorbic acid and the salts thereof, chlorobutanol, benzyl alcohol, thimerosal, phenylmercuric acetate and nitrate, nitromersol, benzalkonium chloride, cetylpyridinium chloride, methyl paraben, and propyl paraben. Particularly preferred are the salts of benzoic acid, cetylpyridinium chloride, methyl paraben and propyl paraben. The compositions of the present invention generally include from 0-2% preservatives.

Preferred sweeteners include, but are not limited to, sucrose, glucose, saccharin, sorbitol, mannitol, and aspartame.

Particularly preferred are sucrose and saccharin. Pharmaceutical compositions of the present invention include 0-5% sweeteners.

Preferred viscosity agents include, but are not limited to, 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. Compositions of the present invention include 0-5% viscosity agents.

Preferred fillers include, but are not limited to, lactose, mannitol, sorbitol, tribasic calcium phosphate, dibasic calcium phosphate, compressible sugar, starch, calcium sulfate, dextro WO 93/24494 PCr/US93/04469~ -24and microcrystalline cellulose. The compositions of the present invention contain from 0-75% fillers.

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

Preferred glidants include, but are not limited to, talc and colloidal silicon dioxide. The compositions of the present invention include from 1-5% glidants.

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

Preferred binders include, but are not limited to, acacia, tragacanth, hydroxypropylcellulose, pregelatinized starch, gelatin, povidone, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, sugar solutions, such as sucrose and sorbitol, and ethylcellulose. The compositions of the present invention include 1-10% binders.

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

Preferably the compounds of the present Invention comprise from about 20% to about 80% by weight of the pharmaceutical compositions of the present invention.

Accordingly, the pharmaceutical compositions of the present invention include from 15-95% of a quaternary nitrogencontaining phosphonate compound active ingredient, or mixture, thereof; 0-2% flavoring agents; 0-50% co-solvents; 0-5% buffer system; 0-2% surfactants; 0-2% preservatives; 0-5% sweeteners; viscosity agents; 0-75% fillers; 0.5-2% lubricants; glidants; 4-15% disintegrants; and 1-10% binders.

Suitable pharmaceutical compositions are described herein in Examples 9 to 11. It is well within the capabilities of one skilled -in the art to vary the non-limiting examples described herein to achieve a broad range of pharmaceutical compositions.

0O 93/24494 PCT/US93/04469 The choice of a pharmaceutical excipient to be used in conjunction with the quaternary nitrogen-containing phosphonate compounds of the present compositions is basically determined by the way the phosphonate is to be administered. If the compound is to be injected, the preferred pharmaceutical carrier is sterile, physiological saline, the pH of which has been adjusted to about 7.4. However, the preferred mode of administering the phosphonates of the present invention is orally, and the preferred unit dosage form is therefore tablets, capsules and the like, comprising from about 0.1 mg P to about 600 mg P of the phosphonic acid compounds described herein. Pharmaceutical carriers suitable for the preparation of unit dosage forms for oral administration are well known in the art. Their selection will depend on secondary considerations like taste, cost, and shelf stability, which are not critical for the purposes of the present Invention, and can be made without difficulty by a person skilled in the art.

The term "mg as used herein, means the weight of the phosphorus atoms present in an amount of a phosphonic acid compound of the present invention. This unit is used to standardize the amount of the phosphonic acid compounds of the present invention to be used in the pharmaceutical compositions and methods of the present inventions. For example, N-(4-hydroxy-4,4-diphosphonobutyl)-N,N-dimethyl-N-(2-mercaptoethyl)-ammonium chloride has a molecular weight of 373.5 g/mole, of which 17% 62 g/mole) is due to the two phosphorus atoms present in this molecule. One milligram of this compound is therefore calculated to have 0.17 mg P. Thus, to prepare a pharmaceutical composition containing 0.17 mg P of this compound, the composition should contain 1 mg of the compound; and to dose 0.17 mg P/kg of this compound to a 50 kg patient, the patient would be dosed with 50 mg of this compound.

The pharmaceutically-acceptable excipient employed in conjunction with the diphosphonates of the present invention is used at a concentration sufficient to provide a practical size to WO 93/24494 PCT/US93/04469 -26dosage relationship. Preferably, the pharmaceutically-acceptable carriers, in total, may comprise from about 0.1% to about 99.9% by weight of the total composition and more preferably from about to about Method for Treating or Preventing Diseases Characterized by Abnormal Calcium and Phosphate Metabolism Another aspect of the present invention is methods for treating or preventing diseases characterized by abnormal calcium and phosphate metabolism. Such methods comprise administering to a human or lower animal in need of such treatment a safe and effective amount of phosphonate compound of the present invention.

The preferred mode of administration is oral, but other known methods of administration are contemplated as well, e.g., dermatomucosally (for example, dermally, rectally and the like) and parenterally (for example, by subcutaneous injection, intramuscular injection, intra-articular injection, intravenous injection and the like). Inhalation is also included. Thus, specific modes of administration include, without limitation, oral, transdermal, mucosal, sublingual, intramuscular, intravenous, intraperitoneal, and subcutaneous administration, as well as topical application.

The term "abnormal calcium and phosphate metabolism", as used herein, means conditions which are characterized by anomalous mobilization of calcium and phosphate leading to general or specific bone loss, or excessively high calcium and phosphate levels in the fluids of the body; and conditions which cause or result from deposition of calcium and phosphate anomalously in the body. The first category includes, but is not limited to, osteoporosis, Paget's disease, hyperparathyroidism, hypercalcemia of malignancy, heterotopic ossification, and osteolytic bone metastases. The second category includes, but is not limited to, myositis ossificans progressiva, calcinosis universalis, and such afflictions as arthritis, osteoarthritis, WO 93/24494 PC1'US93/04469 -27neuritis, bursitis, tendonitis and other inflammatory conditions which predispose involved tissue to deposition of calcium and phosphate.

The term "rheumatoid arthritis" as used herein, means a chronic systemic and articular inflammatory disorder of unknown etiology. It is characterized by destruction of articular cartilage, ligaments, tendons, and bone.

The term "osteoarthritis" as used herein, means a non-inflammatory disorder of the movable joints. It is characterized by deterioration and abrasion of the articular cartilage; and new bone formation at the joint surface.

The terms "person at risk" and "person in need of such treatment", as used herein, mean any human or lower animal which suffers a significant risk of abnormal calcium and phosphate metabolism if left untreated, and any human or lower animal diagnosed as being afflicted with abnormal calcium and phosphate metabolism. For example, postmenopausal women; persons undergoing certain steroid therapy; persons on certain anti-convulsant drugs; persons diagnosed as having Paget's disease, hyperparathyroidism, hypercalcemia of malignancy, or osteolytic bone metastases; persons diagnosed as suffering from one or more of .the various forms of osteoporosis; persons belonging to a population group known to have a significantly higher than average chance of developing osteoporosis, e.g., postmenopausal women, men over age 65, and persons being treated with drugs known to cause osteoporosis as a side effect; persons diagnosed as suffering from myositis ossificans progressiva or calcinosis universalis; and persons afflicted with arthritis, osteoarthritis, neuritis, bursitis, tendonitis and other inflammatory conditions which predispose involved tissue to deposition of calcium and phosphate.

The phrase "safe and effective amount", as used herein, means an amount of a compound or composition of the present invention high enough to significantly positively modify the condition to be treated, but low enough to avoid serious side effects (at a reasonable benefit/risk ratio), within the scope of '0 93/24494 PCI'/US3/4469 -28sound medical judgment. The safe and effective amount of phosphonate compounds of the present invention will vary with the particular condition being treated, the age and physical condition of the patient being treated, the severity of the condition, the duration of the treatment, the nature of concurrent therapy, the specific phosphonate employed, the particular pharmaceutically-acceptable carrier utilized, and like factors within the knowledge and expertise of the attending physician. However, single dosages can range from 0.01 mg P to 3500 mg P, or from 0.0002 to 70 mg P/kg of body weight (based on a body weight of 50 kg). Preferred single dosages are from 1 mg P to 600 mg P, or from 0.02 to 12 g P/kg of body weight (based on a body weight of 50 kg). Up to four single dosages per day may be administered. Daily dosages greater than 500 mg P/kg are not required to produce the desired effect and may produce undesirable side effects. The higher dosages within this range are, of course, required in the case of oral administration because of limited absorption.

The following Examples further describe and demonstrate the preferred embodiments within the scope of the present invention.

The Examples are given solely for the purpose of illustration, and are not to be construed as limitations of the present invention since many variations thereof are possible without departing from its spirit and scope.

Example 1 Synthesis of N-(4-hvdroxv-4.4-diohosohonobutvll- N.N.N-trimethvl ammnium iodide CH3 +1 POH 2 CH- "i-OH r

H

3 P0 3

H

2

CH

3 WO 93/24494 PCr/US93/04469 -29- I. Synthesis of r4-(N,N-dimethvlamino)-1-hvdroxy butylidenelbisprhosphonic acidl A solution containing 4-(N,N-dimethylamino) butanoic acid (2.9 mmol), phosphorus trichloride (2.0 mmol) and diethylphosphite (12 mmol) is stirred 30 minutes at room temperature and then heated at 60C for 24 hours. The reaction mixture is then cooled to room temperature and concentrated hydrochloric acid (50 ml) is added. The reaction mixture is then heated an additional 24 hours at reflux; then cooled to room temperature and filtered through celite and the filtrate is concentrated under vacuum. The crude product is triturated in ethanol, collected by filtration and then dried under vacuum.

II. Synthesis of N-(4-hydroxy-4,4-diDhosphonobutyl)-N,N,N-trimethyl ammonium iodide The bisphosphonic acid (0.30 mmol) is dissolved in water ml) and ethanol (15 ml) and the pH is adjusted to 7.0 by the addition of IN NaOH. To this is added methyl iodide (1.50 mmol) and the reaction is heated at reflux for 24 hours. The mixture is 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 the addition of isopropanol. The product is collected by filtration, rinsed with acetone and then further dried under vacuum.

Example 2 Synthesis of N-(3-hvdroxv-3.3-diDhosohonopropvl)- -N.N-dimethvl-N-oentvl ammonium iodide

CH,

CH(CH2)3CH2-I PO3H I rOH r CH, P0 3

H

2

I

WO 93/24494 WO 9324494PC1Y US 93/04469 I. Synthesis of ('3-(N,N-dimethylamino)pronvlidenelbiscphosphonic acidi Using essentially the same procedure as described in Example 1, part 1, hereinbefore, !J-(N,N-dimethyl amino) propanoic acid is converted to [3,-(N,N-dimethylamino)propylidene]bis- [phosphonic acid].

II. Synthesis of N-(3-hydroxy-3.,3-diohosphonooropyl)-N.N-dinlthylbN-pentyl ammonium iodide The bisphasphonic acid (0.50 ninol) is dissolved in water ml) and acetonltrile (20 ml) and the pH is adjusted to 7.0 by the addition of IN NaOH. To this is added pentyliodide (2.50 mol) and the reaction mixture is heated at reflux for 22 hours. The mixture is then concentrated under reduced pressure and the solid residue is triturated in acetone. The product can then be recrystallized from water and ethanol, 20Exml3 Synthesis of N-(3-hvdroxy-3,3-diphosohonooroovl 1- NN.N-trimethlamionium iodide

CH

3 1+

CH

3 -N OH 2

CH

3 P0 3

H

2 Using essentially the same procedure as described in Examnple 1, part II, hereinbefore, [3-(N,N-dimethylamino)propylidene~bis~phosphonic acid], prepared as described in Example.2, part 1, hereinbefore, is converted to VO 93/24494 PT S9/46 PCT/US93/04469 -31- N-(3-hydroxy-3,3-diphosphonopropyl)-N,N,N.trimethylammonium iodide.

aythess oExample 4 Synhess f N-(2-acetvylthioethyl)-N-(4-hvdroxy-4,4diphosphoflobut-yl )-N.N-djmethyI ammonium bromide

CH

3 +1I P0 3

H

2

C

3

C(O)SC

2

C

2 P&OH2 B

CH

3 [4-(N,N-dimethylamino)-l-hydroxybutylidene~bis[phosphonic (0.75 mmol), prepared as described in Example 1, part 1, hereinbefore, is dissolved in water (50 ml) and acetonitrile ml). To this is added S-acetyl-2-bromoethanethiol (3.75 mmnol) and the reaction mixture is heated at reflux for 12 hours. The is then concentrated under reduced pressure and the solid is triturated in acetone. The quaternized product can be recrystallized from water and ethanol.

of N-(2-acgtylthioethL (3-hvdrgxy-3.3-diohosphonoorgovl 1-N-methyl -N- Dgntyl anmnoniuM bromide

CH

3 CH3(CH 3 CH- S0 O H CH I-OH Bt CH3C()S,) P0 3

H

2 1. Synthesis- of r3-fN-methyl-N-oent DmioloroRvljdenelbis,poi gd WO 93/24494 r/ US93/04469 Using essentially the same procedure as described in Example 1, part I, hereinbefore, 3-(N-methyl-N-pentylamino)propanoic acid is converted to [3-(N-methyl-N-pentylamino)propylidene]bis[phosphonic acid].

II. Synthesis of N-(2-acetylthioethyl)-N-(3-hydroxy-3,3-diphosphonoprovyl)-N-methyl-N-pentvl ammonium bromide Using essentially the same procedure as described in Example 4, hereinbefore, [3-(N-methyl-N-pentylamino)propylidene]bis[phosphonic acid] is converted to N-(2-acetylthioethyl)-N-(3hydroxy-3,3-diphosphonopropyl)-N-methyl-N-pentyl ammonium bromide.

Examle 6 Schenk Model The compounds are evaluated for in yjvi bone resorption inhibition and mineralization inhibition in an animal model system known in the field of bone metabolism as the Schenk Model.

The general principles of this model system are disclosed in Shinoda et al., Calcif. Tissue Int., 5, 87-99 (1983); and in Schenk et al., Calcif. Tissue Res. 11 196-214 (1973), the disclosures of which are incorporated herein by reference.

Materials and Methods: Animals Preweaning 17-day-old (30 gms) male Sprague Dawley rats (Charles River Breeding Laboratories) are shipped with their mothers and placed in plastic cages with their mothers upon arrival. At 19 days of age, pups receiving Rat Chow and water ad libitu are randomly allocated into treatment or control groups comprising seven animals per group. On day 1 and again on day 7 all animals are given an intraperitoneal injection of Calcein solution in 0.9% saline solution; dosed at 0.2 ml/100 g body weight). On day 4 all animals are given an IP injection of tetracycline hydrochloride solution in 0.9% saline '0 93/24494 PC1US93/0469 -33solution; dosed at 0.2 ml/100 g body weight). These compounds label actively mineralizing bone and cartilage.

Dose Solutions and Dosing Procedure All solutions are prepared For subcutaneous injection in 0.9% normal saline and adjusted to pH 7.4 using NaOH and/or HCI.

Dose solution calculation is made by considering the mass of powder (based on molecular weight, hydration) of the active material in mg/kg (body weight) that corresponds to mgp/kg.

Concentrations are based on dosing 0.2 ml/100 g body weight.

Typically, all compounds are administered at 0.01, 0.1, 1.0 and 10.0 mg P/kg/day for 7 days. Compounds showing activity at 0.1 mg P/kg/day are then tested at logarithmic decrements down to 0.001 mg P/kg/day. Adjustments in dosage based on changes in body weight are made on a daily basis.

Necrops,. Tissue Processing and Histomorphometry On day 8 after the start of dosing, all animals are sacrificed by IP overdose of pentabarbitol. Tibias are dissected free and placed in 70% ethyl alcohol. One tibia is dehydrated in graded ethanol solutions and embedded in methyl methacrylate as described in Schenk, Methods of Calcified Tissue Preparation Dickson, Editor; Elsevier Science Publ., The Netherlands; 1984), the disclosures of which are incorporated herein by reference in their entirety. The tibia is sectioned longitudinally through the metaphyseal area. Specimens are stained on one surface with silver nitrate and mounted on microscope slides for evaluation with a Quantimet Image Analyzer (Cambridge Instruments, Inc.) using both incandescent and ultraviolet illumination. Metaphyseal trabecular bone content is measured in the region between the fluorescent label and the growth plate: expressed as percent of total area (bone marrow).

Epiphyseal growth plate width is obtained as the mean value of equally-spaced measurements across the section.

Statistical evaluation of data is made using parametric and non-parametric analysis of variance and Wilcoxons rank sum test to determine a statistically significant effect compared to 0 93/24494 PCT/US93/04469 -34control animals. The Schenk model provides data for in vivo bone resorption inhibition by the compounds.

Example 7 Adjuvant Arthritis Model There are numerous animal models of arthritis, among these is adjuvant-induced arthritis using Mycobacterium butyricum.

This model in a number of ways mimics rheumatoid arthritis in the human (joint swelling associated with cellular and pannus invasion of the joint space, bone resorption, and release of chemotaxic factors and lysosomal constituents into the joint space) A number of prophylactic and therapeutic studies have indicated the potential use of anti-inflammatory drugs (3,4) and diphosphonates in arthritis

REFERENCES

1. Pearson, 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, Burns, Framer, Radziwonik, H., Westwick, J. (977), An X-ray Analysis of Adjuvant Arthritis in the Rat. The Effect Prednisolone and Indomethacin, Agents and Actions, 7:145-151.

3. Winter, Nuss, G.W. (1966), Treatment of Adjuvant Arthritis in Rats with Anti-inflammatory Drugs, Arth.

RheumL, 9:394-404.

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

Francis, Flora, L. King, W.R. (1972), The Effects of Disodium Ethane-1-Hydroxy-1-Diphosphonate on Adjuvant Induced Arthritis in Rats, Calcif. Tiss. Res.., 9:109-121.

WO 93/24494 PC/US931/04469 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 cellulitis and ovitis induced in male rats (either Sprague Dawley or Lewis strain) by a single subcutaneous (SC) injection of Mycobacterium butyricum (8 mg/ml) in mineral oil on day 0. The compounds are dosed once daily either orally (PO) or parenterally (SC) and can be tested in either prophylactic (from day 0) or therapeutic (from day 9 or or 14) protocols. Antiarthritic efficacy can be measured as a reduction in paw volume, body weight loss, bone loss or reactive new bone formati,,. compared to the saline-treated arthritic controls. Treatment can be stopped and the "flare" response (rapid increase in inflammation) examined, which indicates a compound's ability to maintain efficacy.

Materials and Methods A. Animals Animals used are male Lewis rats (LEW). On arrival, the rats are randomized by computer generated random numbers and placed in individual wire suspended cages. Food and water are administered ad libitum, throughout the entire study. Routine care and maintenance of the animals are performed according to State and Federal regulations. Each rat is identified with a number placed in front of the cage and on the tail of the rat.

B. Exaeslmntj~__esian On day 1 body weights (BW) and hind paw volume [(PV) recorded by a mercury displacement method using a pressure transducer linked into a computer] measurements are taken on all rats. On day 0, the induction of arthritis using MFA [Mycobacterium butyricum (Mb) 4.4 mg/kg in oil] is as follows: rats are anesthetized and receive a single SC injection of MFA at the base of the tail under aseptic conditions.

Paw volumes and body weights are measured thereafter on various days, usually twice a week. For the prophylactic WO 93/24494 PC/US93/04469 -36protocol, rats are randomly allocated into groups of 8-10 rats and treatment begins on day 0 and continues daily until termination. For the therapeutic protocol, the rats are randomized into treatment groups of 8-10 rats according to their PV on day 10. Dosing begins on day 10 and continues daily until terminatiton. or both protocols, animals are placed in shoe box cages with deep bedding on or before day Dosing Solutions For Compounds Unlikely to Oxidize Drugs are weighed out on a calibrated balance and then mixed with distilled water in a volumetric flask. The solution is adjusted to pH 7.4 with 0.1N NaOH. Then the solution is filtered through a 0.45 jm sterile filter into a sterile storage container. When not in use, the solution is stored in the refrigerator.

For Compounds Likely to Oidize Drugs are weighed out on a calibrated balance and then mixed with deoxygenated water in a volumetric flask. The stock solution is filtered through a 0.45 pm sterile filter into a sterile storage container. When not in use, the stock solution is kept refrigerated.

On a daily basis, a specific amount of solution is removed from the stock solution, put into small dosing beaker and then adjusted to pH 7.4 according to a predetermined calculation.

Further dilutions of the adjusted solution can be made if necessary (with deoxygenated water).

Drug calculations are made based on the molecular weight, the purity of the compound, the amount based on mg/kg (body weight) and the desired final concentration in mgP/kg. The volume dosed per rat is 0.1 ml/100 gm of body weight subcutaneously, given as an injection in the inguinal fold of the animal, alternating sides each day or 1 ml/200 gm BW given orally using a curved stainless steel dosing tube. Adjustments based on changes in body weight are made weekly.

WO 93/24494 11,CT/LJS93/04469) -37- Radiographs, Necropsy and Tissue Collection At termination, each rat is sacrificed with 1 ml Socomb® intraperitoneally Immediately a whole body radiograph is taken by a Torrox 120D x-ray unit at MA=5, ISUP-50 and second on Kodak non-screen medical film. Hind legs are removed from each rat and fixed in 10% buffered formalin along with a piece of liver, kidney, spleen, and thimus. The tibiotarsal joints are decalcified in 4% EDTA, pH 7.4 and processed routinely in paraffin blocks and H+E stain. The organ parts also processed in paraffin and stained H+E.

The histology sections are evaluated qualitatively for bone and soft tissue lesions using light microscopy. Radiographs are graded for bone resorption (BR) in 6 anatomical trabecular bone sites in each hind leg and 4 sites in each front leg on a scale of 0-3 giving an arbitrary score of 0-60 for all 4 legs. For reactive new bone formation (RNB), radiographs are graded on a severity scale of 0-3 for the lateral and medical surfaces of the tibia and then 0-2 for all other areas mentioned above, giving an arbitrary score of 0-44.

D. Statistical Analysis: Data analysis on paw volume, bone resorption and reactive new bone formation is performed by student's t-test and one-way analysis of variance with Tukeys (SAS) Differences are considered significant at p-0.05 or less.

This model provides in vivo data for the efficacy of antiarthritic compounds in terms of reducing paw swelling bone loss and reactive new bone formation compared to the saline treated arthritic animals.

Example 8 Capsules are prepared having the following composition: Active Ingredient Mq Per Capsule N-(3-hydroxy-3,3-diphosphonopropyl)- N,N,N-trimethyl ammonium chloride 350.0 WO 93/24494 PCr/US93/04469 -38- Excipients Lactose 90.0 Microcrystalline Cellulose 60.0 Magnesium Stearate The capsules having the above composition are prepared using conventional methods as described below, The active ingredient is mixed with the microcrystalline cellulose in a turn shell blender for approximately ten minutes.

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

The mixture is put back into the twin shell blender along with the lactose and is then mixed for approximately fifteen minutes.

The magnesium stearate is next added and blended for an additional five minutes. The resulting blend is then compressed on a piston-activated capsule filler.

Any of the compounds prepared according to Examples 1 to 2 may be substituted for the active ingredient in the capsule prepared hert nabove.

Example 9 Tablets are prepared having the following composition: 25 Active Inredient Mq Per Tablet N-(4-hydroxy-4,4-diphosphonobutyl)- 700.00 N,N-dimethyl-N-(2-mercaptoethyl) ammonium chloride ExciDients Lactose (spray-dried) 200.0 Starch (1500) 100.0 Magnesium Stearate 25.0 Tablets are prepared having the above composition using conventional methods as described below: WO8 9/24494 /2IYCr/US93/04469 -39- The active ingredient is ground in a ball mill for approximately thirty (30) minutes. The milled active ingredient is then blended in a twinblade mixer with the spray-dried lactose for approximately twenty (20) minutes.

The starch is added to the mixture and is then mixed for an additional fifteen (15) minutes. The blend is compressed into tablets on a standard tablet press.

Any of the compounds prepared according to Examples 1 to may be substituted for the active ingredient in the tablet prepared hereinabove.

Examle Injectable solutions are prepared by conventional methods using 10.0 ml of physiological saline solution and N-(4-hydroxy- 4,4-diphosphonobutyl)-N, N, N-trimethyl ammonium chloride, adjusted to pH 7.4.

One injection, one time daily for 4 days, results in appreciable alleviation of rheumatoid arthritis in patients weighing approximately 70 kilograms.

Any of the compounds prepared according to Examples 1 to may be substituted for the active ingredient in the injectable solution prepared hereinabove.

Example 11 A Caucasian male, weighing approximately 92 kilograms, seventy-two years of age, suffering from moderate to severe pain, and occasional swelling, of the right knee. After approximately one year of steadily increasing discomfort, he visits a physician who renders a clinical diagnosis of osteoarthritis of the right knee, which was subsequently verified by X-ray diagnosis.

After a period of ameliorative therapy of various NSAIDs, including aspirin, naprosen, and ketoprofen, his symptoms continue to worsen and his condition appears to degenerate. He returns to his physician who then prescribes the tablets prepared as described in Example 9 twice daily two hours before or after meals for a period of three months. His clinical symptoms of WO 93/24494 IIC'r/ US93/04469 pain and swelling, particularly with extended walking, improved significantly after his 3 months of therapy. At the conclusion of three months at a dosage of 2 tablets per day, the therapy is continued at one-half the dosage originally prescribed 1 capsule, prepared as described in Example 8, per day) indefinitely.

Example 12 A black female, weighing approximately 65 kilograms, fifty-five years of age, presents with swelling and deformation of the finger joints of both hands, with partial loss of strength and/or dexterity of her fingers and hands. Upon visual and X-ray examination and various appropriate clinical tests approved by the American Rheumatological Association (ARA) she is diagnosed with rheumatoid arthritis.

After an unsuccessful analgesic and anti-inflammatory therapy, her physician prescribes the tablets prepared as described in Example 9, two times daily two hours before or after meals for a period of four months. After a month of therapy, her symptoms of knuckle swelling noticeably improves and her range of finger motion increases significantly; she continues therapy for the remainder of the four months, after which her physician continues the prescribed dose for an additional two months.

Example 1 A female of Hispanic origin, twelve years of age, weighing approximately 37 kilograms, presents to the physician with idiopathic juvenile rheumatoid arthritis. Her symptoms include marked inflamation of multiple joints, complicated by heat and tenderness and indicating rapid and pathological degeneration of joint function.

Her physician refers her to a rheumatologist who immediately prescribes aggressive therapy by IV administration of the solution prepared as described in Example 10 over a period of three days, at the rate of 1 injection per day, administered over two hours. At the conclusion of the IV regimen, the physician WO 93/241494 110Y US93/0469 -41prescribes the tablets prepared as described in Example 9, for a period of two months, during which she exhibits marked improvement with increased mobility and decreased pain. For the succeeding two months, the physician reduces her dose to 3/4 of the original oral dose by prescribing 3 tablets over a period of two days, i.e. one 2-tablet day alternating with one 1-tablet day. At the conclusion of this regimen the dosage is again reduced to 1/4 of the original dose by giving her the tablets prepared as described in Example 9, 1 tablet every day for an additional four months.

Example 14 A 60-year-old Caucasian female weighing 62 kg, experiences severe back pain. Her physician, with the aid of a radiologist, diagnoses her as having a crush fracture of the LI vertebrae presumably due to osteoporotic bone loss. The patient is prescribed a three month, once-daily dosage regimen of a 700 mg tablet prepared described in Example 9. The 700 mg tablet is taken either two hours before or two hours after any given meal.

After three months, the dosage is reduced to a 350 mg capsule, prepared according to the procedure described in Example 8, taken every other day for a period of three months. Her physician then puts her on a maintenance dosing regimen wherein she takes a 100 mg capsule, prepared according to the procedures described in Example 8, every day for six months. After six months on the maintenance dosing regimen the patient is not experiencing any further back pain. Follow-up x-rays reveal no additional fractures.

Example A 75-year-old Oriental female weighing 53 kg suffers a fractured hip after a fall. She is hospitalized and diagnosed as having osteoporosis. A treatment regimen of calcitonin injections is prescribed. The calcitonin injections are painful to the patient and she is unable to comply with said calcitonin treatment. Her physician then switches her therapy to an oral WO 93/24494/ P'CIMS93/04469 -42phosphonate regimen. She is administered a 700 mg tablet prepared according to the procedure described in Example 9, twice daily for one month. At the end of this one month of therapy, she is given a 700 mg tablet once daily for two months. At the end of this two month period, she is given a 100 mg capsule daily, prepared according to the procedure described in Example 8, for three months. A follow-up visit to her physician reveals no apparent decrease in mineral density of the forearm as determined by photonabsorptimetry.

Examle 16 A 85-year-old Native American male weighing 65 kg presents to his physician with severe back pain. X-rays reveal multiple minor vertebral body collapse resulting from significant bone loss due to osteoporosis. The patient is prescribed a two month regimen of a 700 mg tablet and a 350 mg capsule to be taken on the same day, eight hours apart, prepared according to the procedures described in Examples 9 and 8, respectively. After two months on this regimen, his dosage is reduced to a 350 mg capsule once a day for two months. X-rays are then taken and an additional crush fracture is noted. He is then put on a maintenance regimen of a 100 mg capsule, prepared according to the procedure described in Example 8, once a day for six months.

At the end of this six months, no significant apparent decrease in bone density is observed.

Claims (12)

1. A quaternary nitrogen-containing compound and it pharmaceutically-acceptable s:;ts and esters thereof having the general formula: RI R 2 /R I 1 P H, R-C N C C-R RIm AR 2 R go o* S wherein m is an integer from 0-10; and n is an integer from 1-10; m n is from 1-10; R is selected from the group consisting of nil; -SR 6 -R 9 SR 6 hydrogen; substituted or unsubstituted C -C 8 alkyl; -OR 3 -CO2R 3 2 -NR 3 2; -N(R 3 )C(0)R 3 -C(0)N(R 3 2 halogen; -C(0)R nitro; hydroxy; substituted or unsubstituted saturated monocyclic or polycyclic .heterocyclic rings; substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic rings; R is selected from the group consisting of -SR S-R9SR 6 hydrogen; substituted or unsubstituted C1-C 8 alkyl; -OR 3 -CO2R 3 -NR 3 2 -N(R 3 )C(O)R 3 -C()N(R3)2; halogen; -C(O)R nitro; hydroxy; substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyclic carbocyclic rings and combinations thereof; -44- each R 2 is selected from the group consisting of substituted or unsubstituted C -C 3 alkyl; -nsubstit4uted o substitutcd phcnylT; benzyl; or R SR; R 3 is selected from the group consisting of H; unsubstituted or substituted C 1 -C 8 alkyl; R SR 6 R 6 i selected from the group consisting of -H; -C(O)R 7 -C(S)R 7 -C(O)N(R 7 2 -C(O)OR 7 -C(S)N(R 7 2 -C(S)OR where R 7 is hydrogen or unsubstituted or substituted C 1 -Cg alkyl; R is selected from the group consisting of -PO 3 H 2 and -P(O)(OH)R where R 4 is an alkyl group having 1-3 carbons. R is substituted or unsubstituted C -C 8 alkyl; R is selected from the group consisting of hydrogen, halogen; SR6; R SR6; amino; hydroxy; substituted and unsubstituted C 1 -C 8 alkyl;

2. A compound according to Claim 1, wherein R 1 is a substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic ring or a substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic ring.

3. A compound according to Claim 2, where R 5 is selected from hydrogen; -SR 6 -R 9 SR 6 substituted or unsubstituted C 1 -C 8 alkyl, .3 -OR 3 -C0 2 R 3 z~ -NR 3 2 -N(R 3 )C(O)R 3 -C(O)N(R 3 2 halogen; -C(O)R nitro; hydroxy; substituted or unsubstituted saturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted saturated monocyclic or polycyclic carbocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyclic heterocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyclic carbocyclic rings and combinations thereof.

4. A compound according to Claim 1, wherein R1 is selected from nil; -SR 6 -R SR 6 hydrogen; substituted or unsubstituted C -C 8 alkyl; -OR -CO2R 3 -NR 3 -R R3; halogen; -C(O)R3; nitro and OH. 0012 00 F-RI 14:27 FAX 01 -1 001-1 3,07r I iP Ci SN1 r HILLUS ORMONDE AIM-COMARWNER la 005 A compound according to Claim 4, wherein R 5 is selected from -SR 6 *,79SR6- hydrogen; suIstituted or unsubstituted CI- C. alkyl; -OR 3 C0 2 R 3 -NR 3 2 -N(R 3 )C(O)fl 3 -C(O)N(R 3 2 halogen; -C(OR ,nitro; hydroxy;, substituted or- unsubstituted saturated monocyclic or polycyJic heterocyclic rings; substituted or unsubstituted saturated ronocyclic or polycyclic carbocyclic rings; substituted or unsubstituted unsaturated monocyclic or polycyc'Iic heterocycli~c rings; substituted or unsubstituted unsaturated monocyclic or polycyclic carbocyclic rings and combinations thereof. A compound according to Claim 4, wherein R 5 is a substituted or unsubstitutei C Ca alkyl. O 7. A compound accvrding to Claim 4, wherein Ris R 9 S 6

8. A compound acc.~ording to Claim 4, whe~rein R5is a substituted or unsubstituted, aturated or unsaturated heterocyclic ring.

9. A compound according to Claim 2, wherein R 5 is d substituted or unsubstitutec, -aturated or unsaturated heterocyclic ring. A pharmaceutical composition comprising: a af and effective amount of a quaternary nitrogen-containing phosphonate COMPOUne, according to Claim 1; and a phirmaceutically-acceptable excipient.

11. A pharnidc(!utical composition comprising: a~ sz fe and effective amount of a quaternary nitrogen- ion~aining phosphonate compound according to Claim 2; a nd pharmaceutically-acceptable excipient,

12. A pharmaceutical composition comprising; 00,12 003 FRI 1; 7 FAX 01 3 001.1 1,807 Il tJPJOM U LOCT(I3NR lJ PUILLIPS ORM(lNUE AIPO-CW(h1I'SS*NCR Q Ooo a safe and effective amount of a quaternary nitrogen-containing p.hosphonate compound according to Claim 4; and a pharmaceutically-acceptable excipient.

13. A pharmaceutical composition comprising: a safe and effective amount of a quaternary nitrogen-containing phosphonate compound according to claim 6; and a pharmaceutically-acceptable excipient.

14. A method for treating or praventing pathological conditions associated with abnormal calcium and phosphate metabolism in humans or other mammals in need of such treatment, comprising administering to said humans or other ***:mammals a safe and effectivc amount of a quaterniary nitrogen-containing phosphate compound having the general -formula: IFIB wherein m is an integer f rom 0-10D; and n is an irltc~er from 1 -10; m n is from 1 R' is selected from the group consisting of nii; -R 8 SR 6 hydrogen; substituted or unsubstitutecid C alkyl; -OR 3 -C0 2 R 3 -NR 3 2 -N(R 3 -C(O)N(R 3 2 halogen; -C(O)W 3 nitro; hydroxy; substitutced or unsubstituted saturated monocyclic or pc;yc -clic heterocyclic rings; substituted or unsubstltuted saturated monocyclic or pulycyolic carbocyclic rings: R5 is selected from- th~e group consisting of 6 -R 9 SR6; hydrogen; substituted or unsubstituted C 1 ,-C 8 alkyl; -OR. 3 -C0 2 R 3 -NR 3 2 -N(R 3 )C(O)R 3 -C(O)N(R 3 2 halogen; -COV:nitro; hydroxy; substituted or unsubstituted saturated monocyclic or polvoyclic heterocyclic rings; substituted or unsubstituted saturate(; nonocyclic or polyc~clic carbocycl ic rings; substituted or u nsubstituted unsaturated monocyclic or polycyclic heterocycilo rings; substituted or 00. 12 '00 FRI 1i 21 AX 031 3 0014 t110PILIP8OMNI+'~Al-~1I8~L 40J PHILLIPS ORMONDE, A]PO-COMMISS"NER Z 0 1) 7 urisubstituted unsaturated rnocyctic or polycyclic carbocyclic rings and combinations thereof; each R2 is selected from the group consisting of substituted or unsubstituted Cl-C33 alkyl; benzyl; or R8r R' Is selected from the gtoup consisting of H; unsubstituted or sJbstituted CI-C 8 alkyl; WSRW; R6 is selected from the group consisting of -C(Q)R 7 -C(S)R- -C(O)Nt.R7)z; -C(O)0R 7 -C(S)N (R 7 2 where R7 is hydrogen or unsubstituted or SUbstituted C- alkyl; R is selected from he group consisting of -PO 3 H 2 and -P(O)(OH)R 4 where R 4 is an alkyl group having 1-3 carbons; R' is substituted or unsubstituted C 1 -Ca alkyl; R 8 is selected from the group consisting of hydrogen, halogen; SR6 R 9 SR'; amino; hydroxy;. ,,,ubstituted and unsubstituted Cj-C8 alkyl.

15. A method liccorciing to claim 14 wherein said humans or other animals are suffering from rheumatoid arthritis, osteoarthritis or osteoporosis.

16. A quaternary nitrogen-containing compound according to claimn 1 substantially as herein described with reference to any one of the examples. DATED: 6 December,'19 THE PROCTER& GAMBLE PHARMACEUTICALS, INC, By their Patent Attorneys PHILLIPS ORMONDE FITZPATRICK INTERNATIONAL SEAIRCH REPORT International Application No PCT/US 93/04469 I-I- I I. CLASSIFICATION OF SUBJECT MATTER (if several classification symbols apply, indicate all)f According to International Patent Classification (IPC) or to both National Classification and IPC Int.Cl. 5 C07F9/38; A61K31/66; C07F9/59 I. FIELDS SEARCHED Minimum Documentation Searched 7 Classification System Classification Symbols Int.Cl. 5 C07F A61K Documentation Searched other than Minimum Documentation to the Extent that such Documents are Included in the Fields Searched s m. DOCUMENTS CONSIDERED TO BE RELEVANT 9 Category Citation of Document, 11 with indication, where appropriate, of the relevant passages 12 Relevant to Claim No 13 X DE,A,4 011 777 (CIBA-GEIGY AG) 1-10 18 October 1990 cited in the application see the whole document X PHOSPHORUS, SULFUR, AND SILICON AND THE 1 RELATED ELEMENTS vol. 54, nr.1-4, 1990 pa- ges 197-202 K.A. JAEGGI A novel rearran- gement in the series of gem-bisphosphonic acids' see page 202 A FR,A,2 319 646 (HENKEL CIE GMBH) 1,9,10 February 1977 see the whole document o Special categories of cited documents to later document published after the international filing date or priority date and not in conflict with the application but document defining the general state of the art which is not dted to understand the principle or theory underlying the considered to be of particular relevance invention "E earlier document but published on or after the international document of particular relevance; the claimed invention filing dat cannot be considered novel or cannot be considered to document which may throw doubts on priority claim(!) or involve an inventive step which Is cited to establish the publication date of another ry, document of particular relevance; the claimed invention citation or other special reason (sa specified) cannot be considered to involve an inventive step when the document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docu- other means ments, such combination being obvious to a person skilled P document published prior to the international filing date but in the art. later than the priority date claimed document member of the same patent family IV. CERTIFICATION Date of the Actual Completion of the International Search Date of Mailing of this International Search Report 19 AUGUST 1993 0 6. 09. 93 International Searching Authority Signature of Authorized Officer EUROPEAN PATENT OFFICE BESLIER L.M. Form PCT/ISA/210 (ucold I ked) (JaIary 195) AN'qNEX TO THE INTERNATIONA SEARCH REPORT ON INTERNATIONAL PATENT APPLICATION NO. us SA 9304469 74283 This annex lists the patent family memnbcrs relating to the patent documents cited in the above-mentioned international search report. The members are as contained in the European Patent Office EDP file on The European Patent Office is in no way liable for theme particulars: which are Merely given for the purpose of information. 19/08/93 Patent document cited in search Meort Publication dateI Patent family member(s) Publication I date DE-A-4011777 18-10-90 None FR-A-2319646 25-02-77 DE-A- 2534391 17-02-77 AT-B- 350161 10-05-79 AT-B- 349642 10-04-79 BE-A- 844649 31-01-77 CH-A- 599234 31-05-78 CH-A- 620359 28-11-80 GB-A- 1540238 07-02-79 JP-C- 1248823 25-01-85 JP-A- 52019628 15-02-77 JP-B- 59025798 21-06-84 NL-A- 7607703 03-02-77 US-A 4054598 18-10-77 jw For more details about this annex see Official Journal of the European Pat Office No. 12/82