FI117053B - New pyridinyl-amino:methylidene bis:phosphonic acid derivs. - used to treat bone disease e.g. osteolytic bone diseases due to malignancy, Paget's disease and osteoporosis - Google Patents

Abstract

Pyridinyl-aminomethylidene bisphosphonic acid tetraesters of formula (I) are new: R1-R4 = 1-5C alkyl; X, Y = H, 1-5C alkyl, halo, OH, 1-5C alkoxy, benzyloxy, acyloxy, NO2, CF3 or NR5R6; R5, R6 = H, 1-5C alkyl or acyl. Also claimed is the use of (I) for the treatment of bone diseases, and a pharmaceutical compsn. contg. (I). In an example, 0.2 moles 2-aminopyridine were mixed with 0.8 mol triethylorthoformate and BF3.OEt2 and the mixt.heated at 150[deg]C for 4 hr. and the EtOH formed was distilled off. Triethylorthoformate was distilled off in vacuum. 0.4 mol diethylphosphite was then added to the reaction mixt. which was heated at 150[deg]C while distilling off the EtOH formed. The mixt. was cooled and the prod. purified to give 29g ((2-pyridinylamino) methylidene)bisphosphonic acid tetraethyl ester.

Description

Pyridyl bisphosphonates for use as a Therapeutic agent

The invention relates to the use of a particular class of pyridylbisphosphonic acid tetraes 5 as a therapeutic agent, er in bone diseases.

Bisphosphonates are therapeutic agents for the treatment of pathologists of various causes, such as lytic bone diseases due to malignant tumor, Paget's disease and oososis. The compounds are physiologically present: analogs of gaseous pyrophosphates. Due to the bisphosphonate P-C-P basic structure, it is possible to exchange a large number of different compounds either! liatom side chains or by adding phosphai

In general, bisphosphonates inhibit bone cancer cells (clasts), which are responsible for bone resorption. Ti 20 bisphosphonates bind to the bone matrix, penetrate into the resorptive bone cancer cells, and reduce bone cell activity. They inhibit bone sores! in vitro and in vivo. Limited absorption from the area, rapid elimination of bone tissue and excretion of unchanged urine are all characteristic of: • 1 * 1 bisphosphate.

• a a • · * * a a a a

The invention is based on the idea of preparing a bisphosphon; a female with high oral bioavailability ·· «: 30 odorous bone affinity. This way you avoid; effect without loss of anti-resorptive active: In patents US 4,447,256, DE 28 31 578 (Suzuki does not · · · · ··· JP 55089210. JP 55098105. JP 55043054. JP 550430! 2

EP 337 706 (Isomura et al.) Discloses the preparation of tetraesters of l- or heterocyclyl-substituted aminomethyl phosphonic acid in which one or more of the ring substituents are saturated. Tetraesters have not been tested.

U.S. Patent No. 4,973,576 to Sakamoto et al. isoxazolyl-substituted aminomethylenylonic acid tetraalkyl esters, but these are the head; 10 tested for arthritis. Their oral bioavailability is low.

EP A 186 405 describes pyridinylamino] methylidene bisphosphonic acids and their esters for use as a medicament for the treatment of bone diseases: Esters are commonly mentioned and no individual inhibitor has been described or tested. Oral] bisphosphonic acids and their salts (bisphosphon; bioavailability is known to be only

EP 282 309 describes azoleaminomethyl phosphonic acids and lower alkyl esters. Tetraei * · '' has not been tested.

EP 325 482 discloses cycloalkylamine <nib bisphosphonic acids and esters. Tetraesters: Tested.

· * • * · • · · · · ·

The invention relates to a number of pyridyl bisphosphonates

Is a novel pharmacological and pharmacokinetic proJ

The present invention relates to pyridylaminomethyl phosphonic acid tetraalkyl esters which are then substituted on the pyridine ring, in particular, the invention relates to methyl phosphonic acid derivatives of general formula I Wherein each of R 1 to R 4 is a straight or branched C 1 -C 5 alkyl group, each of X independently of one another hydrogen, straight or branched C 1 -C 5 alkyl, halogen, hydroxyalkoxy, benzyloxy, acyloxy, nitro, trifluoromethyl or NR 5 R 5 wherein R 5 and R 6 are the same V * and are hydrogen, C 1 -C 5 alkyl or -as> for use as therapeutically active agents v> 25, * X and Y as well as methylene bisphosphonic acid; the amino group of the structure can bind to this position 2-6 of the • · * dyl ring. The groups X and Y are hydrogen or hydroxyl groups, the latter. In this case, one or two hydroxyl groups are preferred

The pyridinyl group is preferably 2-pyridinyl group 4-pentyl, preferably methyl or ethyl. In the definition of an alkoxy group defined by X, the alkyl group has the same meaning as above and is preferably methyl or in the definition of acyloxy as X and Y or R5, the acyl is preferably a lower alkyl group wherein the alkyl group contains 1-5 C mi or ethyl. The groups R 1 -R 4 are preferably the same preferably ethyl.

Preferred compounds of the invention include the following compounds identified: [{2-pyridinylamino) methylidene] bisphosphone] tetraethyl ester, [[(3-hydroxy-2-pyridinyl) amino] methylidene: phosphonic acid tetraethyl ester, [[(6-methoxy-3-) Pyridinyl) amino] methylidene: Tetraethyl ester of phonic acid, 20 [(4-pyridinylamino) methylidene] bisphosphone] tetraethyl ester, [[(5-chloro-2-pyridinyl) amino] methylidene; • · * Formic tetraethyl ester, [[{5-methoxy-2-pyridinyl) amino] methylidene: ♦ · 25 Formic tetraethyl ester, * [[(6-amino-2-pyridinyl) amino] methylidene]] formic acid tetraethyl ester, [[( 3-nitro-2-pyridinyl) amino] methylidene] 1-Phenyl Tetraethyl Ester,: 30 [[(3,5-Dichloro-2-pyridinyl) amino] methylide] ··· * phosphonic acid tetraethyl ester, Λ 5 [[(3-chloro) -5-Trifluoromethyl-pyridinyl) methylidene] bisphosphonic acid tetraethyl ester, [[{2-chloro-3-pyridinyl) amino] methylidene fonic acid tetraethyl ester, 5 [[(6-chloro-3-pyridinyl) amino] methylidene fonic acid tetraethyl ester [ 3-Benzyloxy-2-pyridinyl) amino] methyl bisphosphonic acid tetraethyl ester, [[(5-nitro-2-pyridinyl) amino] methylidene] 10 -onic acid tetraethyl ester, [[(5-benzyloxy-2-pyridinyl) amino] methyl bisphosphonic acid tetraethyl.

N-Substituted (Aminoalkylidene) Bisphosphonic Acid: The esters can be prepared in a known manner by reacting the amino-substituted compound with the aliquotate and reacting the imino ether derivative with the dialkylphos either as such or in the purified form.

In another method, the appropriate aminopyridine is first "reacted with formic acid / acetic anhydride" * · * *. The resulting formamide is then subjected to the treatment of the tetraesters of amino acid pyridine with ··· halomethyl phosphonate by the reaction of re <* phosphorus trihalide and trialkyl phosphite: Y: 25 ββ * Γ Aminoalkylidene bisphosphonic acid. * the diste, after bromination, is reacted with 1; 30 Lifosphite (Schrader et al. Synthesis (1! :: i .: 372).

• · & * 6

The activity of the compounds was confirmed by animal and animal tests, the methods and results of which are given below. In normally growing rats, the type 5 compound, [(2-pyridinylamino) methylidene: phosphonic acid tetraethyl ester, was reduced spontaneously; urinary tetracycline secretions for sorption; chronically pre-labeled rats. The said extract was also effective in preventing bone loss in <10 osteoporosis induced by hip surgery on rats. No effect was observed in the in vitro mouse head capsule culture with calcium release. This may be because: is metabolized before the effects of Farmaco] can be detected. The parent compound also showed no binding to hydroxyapatite in vitro.

The pharmacokinetic properties of the compound [(2-pyridinylamino) methyl] -diphosphonic acid tetraethyl ester were studied in rats. From the intravenous site, Ann <was excreted in urine in small amounts as a parent compound within 2 ^ * · **, which is also an indication of extensive i. About half of the oral compound of this compound was absorbed in rats.

• * · »:: *: The following examples illustrate the invention in a number of ways.

:, ·. Example 1 * * * ***** [(2-Pyridinylamino) methylidene] bisphosphonic acid • · 7 The ethanol formed in the reaction is distilled off: The triethyl orthoformate is distilled off in vacuo. Diethylphosphite (0.4 mol) was heated at 150 ° C when ethanol was distilled off from Form 5. The mixture was cooled and: the product was purified by chromatography (elution: dichloromethane / methanol, 1: 1) to yield 29 g of this product. Physico-chemical characteristics: 10: 31 P NMR (CDCl 3) 15.52 ppm 1 H NMR (CDCl 3): δ 1 ppm ppm protons multiplicity δ: 1.27 δ H m 4.21 δ H m m 4.78 1H d 5.57 1H dt 6.52 1H d CH (6.67 1H m CH (7 , 44 1 H m CH (8.11 1H d CH («1 ti) .V 15 Mass Spectrum (EI Mass):

\ L. 380 M

334 M-EtOH

• · * 243 m- p <o) (oc2h5) 2.

Example 2 • *: [[(5-Chloro-2-pyridinyl) amino] methylidene] bis [beta] 8 during the reaction. Triethyl orthoformate distillate: in vacuo. Diethyl phosphate was added to the reaction mixture; (0.4 mol) and the mixture was heated at 150 ° C to remove ethanol formed ε. The crude mixture was purified by chromatography (eluent: dichloromethane / methanol, 1: 1). There was 26.5 g (32%). (31 P NMR 15.20 ppm; CDCl 3).

Similarly, the following can be prepared: 10 [[(3,5-Dichloro-2-pyridinyl) amino] methyl] -diphosphonic acid tetraethyl ester (31 P-NMR 14.59 ppm [[(3-chloro-5-trifluoromethyl-pyridinyl); methylidene] bisphosphonic acid) tetraethyl ester (31 14.15 ppm; CDCl 3), 15 [[(5-hydroxy-2-pyridinyl) amino] methylidene phosphonic acid tetraethyl ester (mass spectrum (EI 396 M, 350 M-EtOH, 259 M-P (O) (OC 2 H 5)). 2, [[(5-nitro-2-pyridinyl) amino] methylidene] 1-formic acid tetraethyl ester (31 P-NMR 13.97 ppm; C 20 [[(5-benzyloxy-2-pyridinyl) amino] methyl bisphosphonic acid tetraethyl ester, [[ (5-Methoxy-2-pyridinyl) amino] methylidene:: Tetraethyl ester of phonic acid, [[(3,5-Dimethoxy-2-pyridinyl) amino] methyl] bisphosphonic acid tetraethyl ester.

· «· M • ** ·; Example 3 Preparation of tetraethyl ester of [[(3-hydroxy-2-pyridinyl) amino] methylidene] bis acid. . The benzyl 9 mixture was added overnight at room temperature. The reagent was concentrated and the residue was washed with diisopropyl to give 11.4 g of 3-benzyloxypyridinyl-2-formamide. 10 ml of phosphorus trichloride and 1.5 ml of 5 triethyl phosphite were heated at 60-70 ° C. To the solution was added 3-benzyloxypyridyl-2-f '(0.01 mol) and the mixture was stirred for 5 hours at room temperature. The reaction mixture was concentrated and purified by chromatography (eluent: 10 dichloromethane / methanol, 2: 1) to give 0.8 g of [[(: silyloxy-2-pyridinyl) amino] methylidene] bisphos; tetraethyl ester. The benzyl group was hydrogenated, m.p. 0.4 g of [[(3-hydroxy-2-pyridinyl) amino] methylidene bisphosphonic acid tetraethyl ester (Mass Spectrum 15 Mass): 396 M, 350 M-EtOH, 259 M-P (O) (OC 2 H 5). 2.

Similarly, the following can be prepared: [[(6-benzyloxy-3-pyridinyl) amino] methyl] bisphosphonic acid tetraethyl ester, 20 [[(6-hydroxy-3-pyridinyl) amino] methyl ide: phosphonic acid tetraethyl ester.

* · * V 'Example 4 - [[(6-Chloro-3-pyridinyl) amino] methylidene] bis [α] 25 p-tetraethyl ester * ·· *: Diethyl iodomethylphosphonate was prepared as a solution] * · * · Cade, J. Chem. in 1959; 2266.

: 6-Chloro-3-aminopyridine was alkylated with diethyl phosphate 9mL *, ** using sodium amide as the base, and the solution was concentrated in vacuo. The resulting 6-chloro-dinylamino (bromomethyl) -phosphonic acid diethyl (0.1 mole) was heated in tetrahydrofuran with styled phosphite (0.1 mole) at 50 ° C for 4 hours and the reaction mixture was concentrated in vacuo. Purify the product by chromatography (eluting with dichloromethane: toluene, 9: 1). The yield was 5.1 g.

In a similar manner, the following can be prepared: 10 ([(2-chloro-3-pyridinyl) amino] methylidene fonic acid tetraethyl ester, [[(6-methoxy-3-pyridinyl) amino] methylidene fonic acid tetraethyl ester, [(4-pyridinylamino) methylidene] bisf. osion] 15 tetraethyl ester, [[(6-amino-2-pyridinyl) amino] methylidene]] fonic acid tetraethyl ester, [[(3-nitro-2-pyridinyl) amino] methylidene]] fonic acid tetraethyl ester, 20 [[(2- chloro-3-pyridinyl) amino] methylidene] fonic acid tetraethyl ester, [[(5-acyloxy-2-pyridinyl) amino] methyl] * * * phosphonic acid tetraethyl ester.

* ·? Effect on spontaneous bone resorption as assessed for urinary tetracycline secretion in marked rats: * Male Sprague-Dawley rats were used. Starting from the first week, rats were injected subcutaneously with 30 µl of 10 µCi / ml (7-3H (N) tetracycline! ***; taken in physiological saline. Injections, mm λ 11, and feeding to adult animals was commenced on the fifth day. ; randomly grouped and placed individually in: metabolic cages. 24 h 5 urine samples were collected for 10 days. [(2-Pyridinylamino) methylidene; Formic tetraethyl ester of phonic acid, formulated in physiological saline and injected] from day two at different doses. For 6 days Control animals ί 10 saline Urine volume mital The radioactivity of urine samples was determined by n-coil count The results were calculated as the treated roll ratio daily to determine the inhibition of tetracycline secretion.

15

The secretion of unmetabolized tetracycline is a sign of its removal from the bone resorptively during the processes and thus allows for the tracking of the bone orb. As shown in Table 1, inhibition of 20 doses of labeled tetracycline (Compound I) indicates bone marrow. Inhibit.

* · · • · · "1 Table 1 • · · * · *; * 25 ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^:: : avg I 1 mg / kg 5 12.2: 10 mg / kg 5 19.8 ί 100 mg / kg 5 50.0 200 mg / kg 5 72.9 • aa _ ....._______....

12

Male Sprague-Dawley rats weighing 200 + 25 were randomized to weight and grouped using Hypnozm / Mebunat and Temgesic. a physical opening was made to the right or left 5 through which the iliac nerve was exposed and placed within 0.5 cm. The muscle and skin were stitched and returned to their cages. Counterparty foot is left to inexperienced ones. Compound I was dissolved in physiological saline and injected subcutaneously every 10 days at various dose levels starting two before surgery and continuing for 20 days after nerve part removal. Control animals received saline. Animals double-labeled with chromium were sacrificed at standard times 21p after partial excision of the 15 nerve and resected. Thighs were immersed in methyl methacrylate and stained. Methyseal thymus and diaphyseal cortical bone of the thighs were histomorphometrically screened. In control rats, the total bone surface area was reduced in immobilized j As shown in Table 2, dose-dependent increases in thigh bone

• I I

'♦ on an extended foot. Cortical bone mineral · ♦ · "·; no adverse effects * * * 25 were observed on growth rate (results not shown).

• * »• ·· · • · · f · *« 9,999 9 9 9 9 9 9 9 • · '· 30 • · 13

Table 2

Effect on immobilization-induced osteoporosis in rats

Immobilized jc total thigh bone area _% n centers: control __ _ 20 5.3 I 1 mg / kg 4 6.0 37.5 mg / kg 5 17.3 75 mg / kg 5 24.5 __200 mg / kg 7 35 , 5 5

Effect on bone in vitro

Newborn mice were labeled by blue injection of 4SCa 4 days before sacrifice. Fragments of the top 10 were micro-sectioned from the top patches, pre-in culture medium with indomethacin, washed ... after three days, using the binding of the compound of 14 to hydroxyapatite up to a concentration of 500 μΜ (tau 5).

Effect on bone in vitro PTH-stimulated Bone-induced bone loss; inhibition of resorption 100 (PTH-x) /% PTH inhibition ____ (SE) __ I 1 pmole / L no N.D.

5 pmoles / L inhibition did not sit at 10 pmoles / L N.D. ta] 100 pmol / l no 200 pmol / l inhibition bound 500 pmol / l no 1000 pmol / l inhibition bound N.D. no N.D. committed 12.9 (1.3) N.D.

N. D. »Unspecified Pharmacokinetic Droplet: T: 10

The bioavailability was determined within 24 hours from the total amount of urinary compound or serum serum values obtained at various times for oral administration. after intravenous administration. Urine and it. ···. 15 of the samples were analyzed for the Formula I to be tested: * by high pressure liquid chromatography methods.

,,% of the oral dose and 14% of the intravenous <* * * s was excreted as the parent compound within 24 hours (t * * a _____ CO Q- \ T ___ Ί _____- and _____).

Table 4

Excretion of the compound of formula I after a single intravenous or oral dose * AUC0-> co and bioavailability

Excretion 0-24 h AUC0-> ®% of dose F% h * μ9 / ιη1 Ρ · θ · __7.8_ 58 60.7 i-v- 13.6 138.7 5

Dose 114 mg / kg F »bioavailability AUC-> qo - area blood concentration / time curve alli · ·•••••••••••••• · ···· «• · · i *» »· · · ·« • M · · · · · · · · · ···

Claims (8)

1. The tetra-pyridinyl-aminomethylidenebisphosphonic acid tetra of the formula I is 5 γ 9 V-Λ I / OR, (Oh) -nhx / p \ surface V 10 \ p / O R 3 Ϊ XOR 4 0 characterized in that each of the residues R 1 - R 4 is a straight or C 1 -C 5 alkyl group, each of the groups X and Y a: hydrogen, a straight or branched, C 1 -C 5 alkyl group, halogen, hydroxyl, C 1 -C 5 alkoxy, acyloxy, nitro , trifluoromethyl group or N R 5 and R 6 are the same or different and are hydrogen, C 1 -C 5 -, ··· β or -acyl, for use as therapeutic agents. ♦ • · • · · 2. The compound of formula I as claimed in claim 2 as a therapeutically active substance, characterized by the fact that C 1 -C 5 alkyl one in the compound, calculated in the alkoxy and in the acyl group, is methyl ethyl and the halogen is chlorine. : 30 ·· ♦ ·: '** · 3. The compound of claim 1 or 2 for 20 The compound of claim 3 for the therapeutically active substance used, characterized in that the dinyl group is 2-pyridinyl. 5. The compound of claim 1 for therapeutically active substance used, characterized in that the purification is [(2-pyridinylamino) methylidene] bisphosphonic acid ethyl ester, [[(3-hydroxy-2-pyridinyl) amino] methylidene] in the tetraethyl ester of the phonic acid , [[(6-methoxy-3-pyridinyl) amino] methylidene] bisphonic acid tetraethyl ester, [{4-pyridinylamino) methylidene] bisphosphonic acid ethyl ester, [[(5-chloro-2-pyridinyl) amino] methylidene] bispharic tetraethyl ester , [[(5-methoxy-2-pyridinyl) amino] methylidene] bisphonic acid tetraethyl ester, [[({6-amino-2-pyridinyl) amino] methylidene] bisphance tetraethyl ester, C [(3-nitro-2-pyridinyl) ) amino] methylidene] bisphosphoryl tetraethyl ester, [[(3,5-dichloro-2-pyridinyl) amino] methylidene] b tetonic acid tetraethyl ester, ·· · ••• j [[(6-hydroxy-3- pyridinyl) amino] methylidene] bi-phonic acid tetraethyl ester, [[({5-hydroxy-2-pyridinyl) amino] methylidene] bi-phonic acid tetraethyl ester, Li ': [[(3-chloro-5-trifluoromethyl-pyridinyl) amino] m · *** ? the tetraethyl ester of bisphosphonate. 21 [[(3-Benzyloxy-2-pyridinyl) amino] methylidene] tetraethyl ester formic acid, [[(5-nitro-2-pyridinyl) amino] methylidene] bisphonic acid tetraethyl ester or 5 [[(5-benzyloxy-2-pyridinyl) amino] methylidene] tetraethyl ester formic acid. 6. The compound according to some of the above claims for use in the treatment of bone marrow, such as osteolytic bone disease caused by nausea, Paget's disease and primary and secondary os rosis. 7. Pharmaceutical composition for the treatment of 15 judgments, such as osteolytic bone diseases caused. malignancy, Paget's disease and primary and secondary. oporosis, characterized in that the composition pours a pharmaceutically acceptable carrier and an amount of a compound according to any of claims 20 Use of a compound according to any one of claims 1 to 5 as an active agent for dispensing a pharmaceutical composition for b . 1 2 3 f * «·« ·· M »•« · * · 1 · ♦ · »2 • t« 3 · «·· • · • t • M