GB2190836A

GB2190836A - Anti-inflammatories

Info

Publication number: GB2190836A
Application number: GB08711051A
Authority: GB
Inventors: Zvi Dori; David Gershon; Yehuda Scharf
Original assignee: D G S RESEARCH AND DEV
Current assignee: D G S RESEARCH AND DEV
Priority date: 1986-05-13
Filing date: 1987-05-11
Publication date: 1987-12-02
Anticipated expiration: 2007-05-11
Also published as: GB9004351D0; IT8720473A0; AT397036B; CA1306681C; DE3715525C2; IL82376A; DE3715525A1; GB8711051D0; ATA115287A; FR2598616B1; IT1221920B; FR2598616A1; GB2190836B

Description

1 GB 2 190 836 A 1 SPECIFICATION combination of the complexes can be

administered subcutaneously or even topically in a suitable vehicle, Anti-inflammatory substances 65 e.g. physiological saline in the case of s.c. administra tion and dimethyisuifoxide (DIVISO) in the case of a The present invention relates to anti-inflammatory topical administration, although ointments, salves or substances forthe treatment of mammalian subjects like conventional vehicles may be employed.

for inflammatory conditions and, more particularly, For oral administration, the complex or mixture of arthritis and like active-oxygen or superoxide based 70 complexes may be prepared in suitable dosageforms.

conditions. Forexample it may be prepared as dragees, as It has been recognised forsometimethat inflamma- capsules, astablets, as an elixiror other oral dosage tion in mammalian species can betraced at least in fo rm.

partto active oxygen species, including superoxide, The dose may be administered oneto sixtimes and radicals associated therewith atthe inflammatory 75 daily, depending upon the severity of the inflamma site. Considerable research has been undertaken to tory condition, preferably under medical supervision measure and detect oxygen radicals, to establish the so thatthe dosage can be reduced orthe number of mechanisms whereby enzymes such as superoxide daily administrations limited as the inflammatory dismutase is effecteve in countering oxygen radical condition subsides.

toxicity and even in the developmentand use of 80 The compounds of the invention may also have copper amine oxidases in preventing tissue damage prophylactic properties in preventing the spread of and even in promoting damaged-tissue recovery. arthritic inflammation and has been found to be However, the compounds which have been de- effective in reducing the severity of the actual condi veloped heretofore for active-oxygen or superoxide tion which develops in subjects who are prone to such antagonism and destruction in vivo have not proved 85 inflammatory states. The compounds may also be as effective as desired orwere characterised by side effective in preventing postischemic heart damage reactions or could not be made in commercially and for geriatfic applications otherthan as antiarthri significant quantities at reasonable cost. tics.

The substances to be described hereinafter also We have found thatthe anti-inflammatory effect is provide an improved method of treatment of mamma- 90 exceptionally pronounced with low-toxicity metallo lian subjects for inflammatory conditions wherebythe organic complexes selected from the group which aforedescribed drawbacks are obviated. consists of:

The substances to be described hereinafter also [Co{2,3,9,1 0 tetraflower - alkyi} - 1,4,8,11 - tet provide an improved method of treating a mammalian raazacyclotetradeca-1, 3,8,10-tetraene}C121CI, subjectfora condition resulting from active oxygen or 95 a 1,1 dimethylferroceniurn or other alkyl ferrocenium superoxide toxicity which may result in acute or salt, chronic inflammation orany other disorder. a cobalt (111)-bis(acetyl or propio acetone)-ethylene The method of treatmentwhich is nontoxic and has diiminecomplex, no measurable side-effects butwhich is particularly and effective in thetreatmentof arthritic conditions orthe 100 [Co{2,12 - dimethyl - 3,7,11,17 - like. tetraazabicyclo[l 1.3.1 1heptadeca - 1(17),2,11,13,15 According to the present invention there is provided -pentaene}C121CI.H20.

an anti-inflammatory substance fortreating an in- In the case of the ferrocenium derivatives, we have flammatory condition in a mammalian subject, com- discovered that the key to the effect is the ferrocenium prising at least one metallo-organic complex capable 105 ion, so that any physiologically compatible or phar of in vivo su peroxide anti-oxidant effects and having a maceutically effective saitthereof can be employed.

metal selected from the group which consists of Furthermore, the methyl groups are important only in cobalt andiron, in combination with a carrier. the sense that two methylene structures -CH2 According to the present invention there is further moieties are attached to the ferrocenium moiety. The provided the use of compound comprising at least one 110 methyls can thus be substituted by Cl to C5 -alkyl, metallo-organic complex capable of in vivo superox- alkenyl or alkoxy. Best results are obtained wth ide anti-oxidant effects and in which the metal of said 1,1 - dimethylferroceniurn salts.

complex is selected from the group which consists of In the case of the (Co{2,3,9,1 0 -tetra{lower- alkyl} - cobalt and iron in the manufacture of a medicament 1,4,8,11 -tetra azacyc 1 otetrad eca - 1,3,8,10 - forthe treatment of an inflammatory condition in a 115 tetraene}C]2)CI, we have found thatthe lower alkyl can mammalian subject. be Cl to C6-alkyl butthat best results are obtained Substances embodying the invention will now be whenthe described byway of example with reference to the (Co{2,3,9,10-tetra{iower- aikyi}-1,4,8,11-tet- following Examples. raazacyclotetradeca - 1,3,8,10 -tetraene}CIACI is The method is used in thetreatmentof acute or 120 Co{2,3,9,10 - tetramethyl - 1,4,8,11 -tetraazacy chronic arthritis in a dosageof 0.1 to 250 mglkg of clotetradeca - 1,3,8, 10 -tetraene}C]2)CI.

bodyweight, butin all cases at most50% of the LD50 Preferablythe cobalt(iii)-bis(acetyl or propio ace value of the compound,when the complex is adminis- tone)-ethylenediimine complex is a cobalt(I11) tered orally as is preferred. Howeverthe complex or a bis(acetylacetone)ethylenediimine complex, and Formulae in the printed specification were reproduced from drawings submitted after the date of filing, in accordance with Rule 20(14) of the Patents Rules 1982.

2 GB 2 190 836 A 2 more specifica Ity the standing was recrystd. from ethanollwater (com [Co(bis{acetylacetone} - ethyleneidimine) (NH3)21 pound 23).

cl-. Anal. Calcd. for C12H2402N4COCI C, 41.09; H, 6.90.

We have found thatthe complexes specifically Found: C, 40.8 1: H, 7.00.

3 identified above can be used togetherwith hitherto- 55 [Co(CR)021C1.H20 (Compound39) known anti-inflammatories with propionic acid side CR = 2,12 - dimethyl - 3,7,11,17 - chains, especially indomethacineto further alleviate tetraazabicyclo[II 1. 3.1] heptadeca - 1(17),2,11,13,15 the suffering of arthritic inflammation. -pentaene Preparation of the Compounds The Co(I1) complex, [Co(CR)C12Lwas prepared and [Co(TiM)C121CI Compound 8) 60 oxidized bythe procedure described by Poon et al. for Tim = 2,3,9,10 -tetramethyl - 1,4,8,11 -tetraazcyc- the analogous perchlorate (J. Chem. Soc. Dalton, lotetradeca - 1,3,8,10 -tetraene. 1977,1247-1251). The complex (compound 39) was The procedure of Bush et al (Inorg. Chem., 1972, 11, recrystallized from acetone.

2893) was employed to prepare [Co(Tim)CIAPF6. Anal. Calcd. for: C161124ON4COCI3: C, 42.35; H, 5.33; A saturated solution of tetramethylammonium 65 N,12.35; C1,23.45. Found: C,41.35; H,5.69; N,12.07; chloride in acetonewas added slowly to a saturated Cl, 23.64.

solution of [Co(TiM)C121PF6 in acetone until the In vivo Inflammatory Studies solution became cloudy. Cooling afforded light green The drugs used are prepared just before performing crystals of compound 8. the experiment. Drugs are dissolved at a concentra- Anal. Calcd. for C14H24N4COCI3: C,40.63; H, 5.8; N, 70 tion of 1 O-IM (or 2 x 10-2M or4x 10-2M) in pyrogen 13:54. Found: C, 40.57; H, 5.67; N, 13.15. - free sterile saline. The dissolved drug isthenfiltered 1,1 -Dimethylferrocenium Salts (PR6 Cl-, through a sterile and pyrogen- freeO.2 micron filter BF4 -, Br-, C104 -) - Compound 19) (acrodisc, Gelman).

The procedure of Wahl eta]. (J. Phys. Chem., 1975, Procedure 79,2049-52) was employed. 75 8-12 CD-1 female mice (Charles river), age 2- 5 A solution of dimethy[ferrocene (0.65 g. 3 mmol) in months are numbered, weighed and distributed to 2-3 concentrated sulfuric acid (8 mi) was stirred at room cages, 3-4 mice in each cage. 0.2 mi of pyrogen-free temp. for 10 min. and then poured into 60 mi. of cold, saline or drug are injected subcutaneously in a dist.water. The insoluble material was removed by randomized order.

filtration,the deep-blue filtrate treated with 0.82 g (5 80 Thirty minutes after injecting the drugs orthe saline, mmol) of ammonium hexafluorophosphate, stirred the right paw of each mouse is injected with 25 for 112 hr. and cooled. The separated solid (compound microliters of 1 % carrageenin (viscarin type, Marine 19) was collected, washed with ethyl ether and dried Colloids) in pyrogen- f ree saline orwith 5 microliters undervacuum fortwo days. (0.227 U) of xanthineoxidase (Sigma).

Anal. Calcd. for C12H14FePF6: C, 40.14; H, 3.93. 85 1.5 hr to 2 hrs after injecting the inf lammatory Found: C, 39.98; H, 4.23. stimulus to the right paw, both paws of the animal are Cobalt(I11) Bis(acetylacetone)-ethylenediimine Com- amputated atthe knee joint and weighed. The plex (Compound 23) uninjected left paw serves as an internal control forthe [Co(BAE) (NH3)2]CI degree of swelling of the right paw in each animal.

This complex was prepared by a modification of the 90 In some experiments the surface temperature of the procedure described byG. Costa etal.,J. Organomet- right and left paws were recorded also before the al. Chem., 1966,6,181 -187. amputation was performed.

Bis(acetylacetone)-ethylenediimine (BAE, 2.24 g ' 10 Calculations mmol) was dissolved in methanol (100 m]) and treated The difference in mg between the weight of the right with cobalt chloride hexahydrate (2.38 9, 10 mmol). 95 and left paw in control animals (injected with saline) The brown solution was stirred at room temperature represents 100% of the acute inflammatory response.

forl 112 days in order to allow oxidation of the cobalt Concomitantly, the difference between paws of the (no precipitate formed), treated with 7 m] of conc. aq. drug treated animals is calculated and compared to ammonia solution, and then heated under reflux for 2 control.

hrs. The yellow-brown solid that separated upon TABLE 1: Effect of several products on carrageenin paw oedema in mice measured by paw weight.

Paw Weight Product No. of total route of average % % expts. mice admin. mglkg activity inhibition saline - - S.C. - 100% 0 23 4 14 S.C. 25 63.1% 36.9% 8 4 15 S.C. 29.8 73.2% 26.8% 39 5 18 S.C. 35.7 64.7% 35.3% 19 1 4 S.C. 23.5 88.2% 11.8% 19 3 11 S.C. 58.9 54 % 46 % subcutaneously 3 GB 2 190 836 A 3 TABLE2: Effectof several products on xanthineoxidase paw oedema measured bytemperature reduction.

Product No. of total route of average % expts. mice admin. mglkg activity inhibition saline - - S.C. - 100% 0 23 2 6 S.C. 22.7 31 % 69 % 8 3 10 S.C25.1 33.2% 66.8% 19 1 4 S.C. 46.8 69.5% 30.5% 19 3 12 S.C. 91.5 49.6% 50.4% subcutaneously TABLE3: LD50 and ED50 of various drugs on xanthine 40 Example 3 rng. of active ingredient (one or more of oxidase are carrageenin paw oedema in mice om4tlm6t2m5t3m5 compounds 8,19,23 or 39) are dissolved in 1000 mi.

of distilled water. The solution isfilled into 500 stimulus:xanthine carrageenin ampoules. In this way ampoules containing 2 m[. of a oxidase measurement:temp. weightmg 45 solution containing 25 mg.lmi. of active agent each Drug LDo ED50 ED50 are obtained. The contents of an ampoule are injected mglkg mglkg mglkg subcutaneously.

23 75 <38 25 Example 4 19 >>875 93 65 Gelatin capsules of the following composition are 39 85 - 43 50 prepared:

8 172 <24 not active ingredient (one or more determined of compou nds 8,19,23 or 39) 25.0 mg.

maize starch 122.0 mg.

colloidal silica 3.0 m g.

Orally and S.C. Administerable Compositions 55 150.0 mg.

Example 1 The ingredients are homogenized, and the The composition of tablets is as follows: horn-ogenate is filled into hard gelatine capsules.

active ingredient (one or more 1000 capsules of 150 mg. (filling) weight each, of compounds 8,19,23 or 29 25.0 mg. 60 containing 25.0 mg. of active ingredient per capsule, corn starch 97.0 rng. make a lot.

Claims

lo polyvinyl pyrrolidone 175.0 mg. CLAIMS magnesium stearate 3.0 mg. 1. An

anti-inflammatory substance fortreating an inflammatory condition in a mammalian subject, 300.0 mg.65 comprising at least one metallo-organic complex Theactive ingredient and the corn starch are capableof in vivo su peroxide a nti-oxida nt effects and wetted by an aqueous polyvinyl pyrrolidone solution having a metal selected from the group which of approx. 15% w/V1, followed by granulation, and consistsof cobaltand iron, in combination with a drying of the wet granules at about 40-45'C. The dried carrier.

granulate is thoroughly mixed with magnesium, 70
2. A substance according to Claim 1 wherein said stearate, and the mixture so obtained is further metallo-organic complex is selected from the group processed by a tablet machine, equipped with an which consitst of:

appropriate pressing tool, to give tablets of 300 mg. [Co{2,3,9,1 0 tetraflower- alkyl} - 1,4,8,11 -tetra - weight containing 25 mg. of active ingredient. One azacyclotetradeca -1 3, 8,10 -tetraene}C121CI, manufacturing lot includes 1000 tablets. 75 a 1,1-dimethyiferrocenium salt, Example 2 a cobait(iii)-bis(acetyl or propio acetone)-ethylene Dragees of thefollowing composition are pre- diiminecomplex, pared: and active ingredient (one or more [Co{2,12 - dimethyl - 3,7,11,17 - of compounds 8,19,23 or 39) 50.0 mg. 80 tetraazabicyclo[l 1.
3.1 1heptadeca 1(17),2,11,13,15 lactose 94.0 mg. -pentaene}C]2]CI-H20.

polyvinlypyrrolidone 154.0 mg. 3. A substance according to Claim 2 wherein said magnesium stearate 2.0 mg. 1,1-dimethy[ferrocenium salt is 11,11dimethylfer rocenium hexa-fluorophosphate.

300 mg. 85
4. A substance according to Claim 2 wherein said Granulates are prepared according to Example 1, [Co{2,3,9,1 0 -tetra {lower-alkyl} 1,4,8,11 -tetra - and from them dragee kernels of 150 mg. weight are azacyclotetradeca - 1, 3,8,10 -tetraene}CIACI is pressed. The dragee kernels are coated with a layer [Co{2,3,9,11 0 - tetramethyl - 1,4,8,11 -tetraazacy containing sugar and talc followed by coloring with clotetradeca - 1,3,8, 10 -tetraene}C121CI.

an approved food colorant and polishing with bees 90
5. A substance according to Claim 2 wherein said wax. cobalt(] 10-bis(acetyl or propio acetone)-ethyleneidi- 4 GB 2 190 836 A 4 mine complex is cobalt(111)-bis(acetylacetone) - Printed in the United Kingdom for Her Majesty's Stationery Office by the ethylenediimine complex. Tweeddale Press Group, 8991685, 12f87 18996. Published at the Patent Office,
6. A substance according to Claim 2 wherein said 25 Southampton Buildings, London WC2A lAY, from which copies may be cobalt(I11) - bis(acetylacetone) -ethylenediimine com- obtained.

plex is [Co(bis{acetylacetone} -ethylenediimine) (NH3)2]CI.
7. A substance according to any preceding claim wherein the carrier comprises indomethacin.
8. An anti-inf lammatory substance for treating a mammalian subject substantially as hereinbefore described.
9. The use of compound comprising at least one metallo-organic complex capable of in vivosuperoxide anti-oxidant effects and in which the metal of said complex is selected from the group which consists of cobalt and iron in the manufacture of a medicament forthe treatment of an inflammatory condition in a mammalian subject.
10. The use according to Claim 9 wherein said metallo-organic complex is selected from the group which consists of: [Co {2,3,9,1 0 -tetra flower-a 1 kyl} - 1,4,8,11 -tetraazacyclotetradeca - 1, 3,8,10 -tetraene}C121CI, a 1,1-dimethyiferrocenium salt, cobalt(iii)-bis(acetyl or propio acetone)-ethylenedilmine complex, and [Co{2,12 - dimethyl - 3,7,11,17 tetraazabicyclo[l 1.3.1]heptadeca - 1(17), 2,11,13,15 -pentaene}C121CI-H20.
11. The use according to Claim 10 wherein said 1,1-dimethyiferrocenium saltis 1,1-dimethyiferrocenium hexafluorophosphate.
12. The use according to Claim 10 wherein said [Co{2,3,9,1 0 -tetra {lower- alkyl} - 1,4,8,11 - tetraazacyclotetradeca-1, 3,8,11-tetraazacyclotetradeca -1,3,8,10-tetraene}C]2]CI.
13. The use according to Claim 10 wherein said cobalt(111)-bis(acetyl or propio acetone)-ethylene- diimine complex is cobalt(Iii)-bis(acetylacetone)ethylenediimine complex.
14. The use according to Claim 10 wherein said co ba lt(l 10-bis(acetyl acetone) - ethylenediimine cornplex is [Co(bis{acetylacetone}-ethylenediimine) (NH3)2]CI.
15. The use according to Claim 10 wherein said inflammatory condition is arthritis.
16. The use according to Claim 15 wherein said inflammatory condition is acute arthritis.
17. The use according to Claim 10 wherein said metallo-organic complex is administered to the subject in combination with a pharmaceutically effective amount of indomethacin.
18. The use according to Claim 10 wherein said metallo-organic complex is in a form suitable to be administered to a subject in an oral dosage form.
19. The use according to Claim 10 wherein said metallo-organic complex is in a form suitable to be administered to the subject in a topical form.
20. The use of a compound according to anyone of Claims 9to 19 in the manufacture of a medicament forthe treatment of an anti-inflammatory condition in a mammalian subject.