NZ199546A - Lysine 1-benzylindazol-3-yloxyacetates;occular formulations for the treatment of cataract - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number 1 99546 199546 >\ ■ Piioi.'V .

Complete Xpecifocation FiledP?~.'l Class: A?nV?£j. '/£?A<T... Publication Dsto: .5 J, !tAX P.O. Journal No: r»k «*&, Si ty% ■<*?£ Patents Form No. 5 .. iv- NEW ZEALAND PATENTS ACT 1953 COMPLETE SPECIFICATION (i-BENZYL-lH-INDAZOL-3-YL)OXYJ ACETIC ACID SALT WITH LYSINE" HE AZIENDE CHIMICHE RIUNITE ANGELINI FRANCESCO A.C.R.A.F. S.p.A, an Italian company, of Viale Amelia, 70, Rome, Italy, hereby declare the invention, for which i-/we pray that a patent may be granted to -rae-/us, ,and the method by which it is to be performed, to be particularly described in and by the following statement:- 199546 In the following New Zealand Patent Specification No. 151247 a series of compounds is claimed belonging to the general formula wherein X = H, CI, OCH3, NC>2, NH3 or NHCOCH3; and R = H, phenylybenzyl, or phenyl or benzyl substituted with methyl, methoxy, halogen, trifluoromethyl, or dimethylsulfamido and possessing interesting pharmacological properties. Particularly [(l-benzyl-lH-indazol-3-yl)oxy] acetic acid (bendazac) was found to be very interesting as a local anti-inflammatory agent and it was introduced in medicine and used for this activity.

The [(lH-indazol-3-yl)oxy] alkanoic acids particularly [ (l-benzyl-lH-indazol-3-yl)oxy] acetic acid, are only slightly absorbed, even as a salt with alkali metals, alkali earth metals or with organic bases such as morpholine, diethanolamine, piperazine, triethanolamine, diisopropylamine,or 4-(2-hydroxyethyl) morpholine. The bendazac blood concentrations are found in any case to be too low or erratic to permit the systemic use of the drug. 109546 As reported by Silvestrini, Catanese and Lisciani in*"Proceedings of an International sym-' posium on Inflammation Biochemistry and Drug Interaction" Excerpta Medica International Congress Series n° 188 (1968) - the mean serum concentration found in man after an 8-day treatment with daily doses from 150 to 900 mg is only 8 fig/ml.

It has now been found that 1-benzyl-lH-indazol-3-yl)-ox^/acetic acid (bendazac) as a salt with a lysine has a better absorption. As shown in Table I, after a single oral administration of 500 mg of a lysine salt, corresponding to about 300 mg of l_(l-benzyl-lH-indazol-3-yl)-ox£/acetic acid, the mean serum concentration found in 6 volunteers, two hours after administration, is SO^jug/ml; under the same conditions, the blood .concentration obtained with 300 mg j_(l-benzyl-lH-indazol-3-yl) — ox^/acetic acid or bendazac is 8 ji'g/ml.

The availability of a bendazac salt (i,e.lysine salt) capable of producing higher blood concentrations than those previously obtained with bendazac gave results which dramatically modify the drug's possible uses. v-? P , . ^- .'La'U' # • TADt.E T Serun; concentrations in man after a single oral administration of bendazac as such or bendazac.salt with L-lysine.

Compound Sex "Weight in kg Dose in mg Concentrations in jig/ml 1 hr 2 hr 4 hr 8 hr bendazac g ? 50 70 80 60 50 40 300 6 7 2 8 3 9 16 9 11 12 8 7 6 9 10 8 5 4 4 4 X + SE .2 j-0.94 1.1.7 ± 1.26 8.7 + 0 .88 .0 + 0 .63 bendazac salt with /.-lysine le, X + SE 60 55 80 75 60 45 500 [ (equiv. to about 300 of acid) 20 18 23 •20 15 56 32 37 46 27 22 I 6 18 10 II 18.5 + l .28" 36.7 + 5.13" 18.3 ± 3.21 8 9 16 5 5 8.8 + 1.66" 'O QJI 139546 Cataract is a disease of the eye involving opacification of the lens, a bioconvex body interposed between the anterior chambers and the vitreous body of the eye. The lens has the function of focusing the light rays on the retina where they are perceived and transmitted to the brain in the form of visual impulses. Cataract is, therefore, accompanied by a decrease in visual capacity, which in the more severe cases leads to blindness.

The lens is surrounded by a collagenous elastic capsule containing fiber cells whose main component is a homogeneous, transparent protein gel which has the same function as the glass in an artificial lens. With increasing age, or due to a variety of different factors, the lens fibers become opaque and loose their transparency. Until recently these changes were considered irreversible. It was generally believed that the only possible cure for cataract was surgical removal of the lens which was then substituted with external or internal artificial lens implanted behind the eye.

It has now surprisingly been _found, according to the present invention that the oral administration of a therapeutic amount of a bendazac salt, with L-lysine, not only delays cataract development but also actually causes regression. _ 6- 1:39546 The present invention provides the use of bendazac lysine salt having the formula: OCH COOH , 2 ^2-^2 CH2 I 2 COOH for the treatment of cataract, especially in the form of pharmaceutical compositions for oral administration or for administration topically to the eye: the compositions should, of course, provide effective tissue concentration of a lysine salt of bendazac.

Study on the anti-cataract activity in humans A first experiment was conducted on 20 patients with different types of cataract (cortical, corticonuclear, posterior subcapsular). The patients' medical history showed that their cataract was rapidly progressing. The experiment included an examination of vision, refraction and slit-lamp test; the investigator was asked to express an overall judgement. These evaluations were performed before the beginning of treatment, after 4 weeks and again at the end of treatment when prolonged. A lysine salt 109546 bendazac was administered at the dose of 1500 mg « (corresponding to 911.5 mg of bendazac) divided into three daily administrations during meals. The average duration of treatment was 50.A days with time limits from 28 to 86 days. The study group consisted of 11 females and 13 males with an average age of 59.5 (47 - 74).

The effects of treatment are summarized in Table 2: the values obtained in each patient are illustrated instead in Table 3. For practical purposes the results are expressed as: improvement (+), no change (0), worsening (-).

TABLE 2 Summary of the results obtained in the first experiment conducted with a lysine salt of bendazac in cataract Evaluation Vision Refraction Slit-lamp and/or Transillum Over all Judgement improvement (+) 18 14 18 no change (0) 6 13 worsening (-) 1 23^!;U4 TABLE 3 109546 Details on each patient used in the first experiment conducted with a lysine salt of bendazac in cataract Name Age Sex Type of cataract Days treatment Vision Refraction Slit- lamp and/or transil-lumin Overall judgement Note V.C. 47 F subcapsular 63 + 0 + + S.E. 51 F cortical 28 + + + + I.A. 53 F cortical 78 + + + + F.A. 55 M cortical 33 + + + + D.M. 56 M corticonuclear 84 + 0 + + D.A.P. 58 M corticonuclear 72 + + 0 +, A.M. 63 F cortical 79 + + 0 + A.G. 65 M subcapsular 0 I.D. 66 M subcapsular 34 + 0 + + A.C. 66 F subcapsular 33 + + + + C.M. 67 F subcapsular 65 + 0 + + C.L. 67 M cortical 54 + + 0 + T.A. 68 M cortical 31 + + 0 + i.e. 68 F corticonuclear 61 + + + + P.A. 69 M cortical 0 0 + 0 L.G. 69 M subcapsular 86 + 0 + + C.R. 73 F cortical 49 + + + + L.G. 73 M subcapsular 73 0 + + + L.C. 51 M subcapsular 54 + 0 0 0 Q. A. 73 F cort.-nucl. 29 + + 0 + L.E.N. 58 M subcapsular 0 0 0 0 diabetes F.G. 74 F cort.nucl. 72 0 0 0 0 II L.E. 50 F subcapsular 29 0 + 0 0 II E.P. 63 M subcapsular 29 + 0 + + If No. cases 1 2 3 4 7 8 9 11 12 13 14 16 17 18 19 21 22 23 24 2r3JWri984 109546 \ In this experiment a lysine salt pf bendazac t produced an improvement in a high number of patients (10 - 18, depending on the evaluation method), whereas only 1 patient showed a worsening effect. Relatively poor results were obtained in 5 patients, one of whom was a diabetic.

The experiments were repeated under double blind conditions comparing a lysine salt of bendazac with placebo. A total of 35 patients were used; 19 were treated with placebo and 16 with L-lysine salt of bendazac, using the same experimental methods except that the duration of treatment was always A weeks. The results are summarized in Tables 4 and 5.

TABLE 4 Summary of the results obtained in the second experiment conducted with the double blind method to study the activity of a Lysine salt of bendazac in cataract .1 Table 4 No. Cases r*" ■1 1 - - Sex Type of Cataract Treatment Evaluation Vision Refraction Slitlamp Transillumination Overall Judgement ■ 12F Cortical 13 (+) 1 1 _ _. 1 19 7M Corticonuclear 6 .

Placebo (0) (-) 6 12 12 6 2 9 1 6 2 16 16 ' 14F 2M i Cortical 13 Corticonuclear 3 Bendazac L-lysine salt (+) (05 (-) \ V 6 9 7 2 1 4 2 " _ 11 23 i 2 i • • "11_ 1:19546 TABLE 5 Details on each patient used in the second experiment conducted with the double blind method to study the activity of lysine salt of bendazac in cataract.

No. of Name Sex Type of Treatment Vision Refrac Slit- Transil- Overall cases cataract tion lamp lumin judgement 1 P.A.

F corticonuclear placebo 4 L.G.C.

M cortical placebo + + 0 + <r M.M.

F cortico placebo - - - — nuclear 8 F.P.

F cortical bendazac L 0 0 0 9 L.O.

F corticonuclear placebo — M.M.

F corticonuclear bendazac L + 0 + + + ^11 S.M.

F cortical placebo 0 0 0 mi2 C.P.

M cortical placebo 13 L.V.

F cortical bendazac L + 0 + + 14 S.K.

M cortical placebo - F.F.

F cortical bendazac L 0 0 0 0 0 16 G.A.

F cort.nucl. bendazac L 0 0 - - 17 P.E.

F cortical bendazac L + + + + 18 C.G.

F cort.nucl. placebo - - 19 A.R.

F cortical bendazac L + + + + T.D.

M cort.nucl. placebo - - - 21 C.L.

M cortical placebo 23 C.I.

F cortical placebo - - 24 C.L.

F cortical bendazac L 0 + + + A.A.

F cortical placebo 0 0 . - - - 26 V.F.

M cortical placebo 27 D.S.E.

F cortical bendazac L + + + + 28 F.C.

M cortical placebo 0 0 0 0 0 29 M.R.

F cortical bendazac L 0 + + + + M.A.

M cort.nucl. bendazac L + 0 0 0 0 31 T.A.

F cortical placebo 0 - - - - 32 L.C.

F cortical placebo 0 0 - - - k 33 D.A.L.

F cortical bendazac L + + + + + m::.4 G.A.

F cortical placebo 0 - - - W 35 E.D.

F cortical bendazac L + + + + 36 P.A.

F cortical bendazac L 0 0 - - 37 C.M.

F cortical placebo - 38 A.M.

M cortical bendazac L + + + + 39 V.P.

F cort.nucl. placebo - - - 40 P.E.

F cortical bendazac L + + + + * 199546 These studies confirm without doubt that the administration of bendazac L. Lysine salt has a healing effect on cataract.

Finally eye drops containing 0.2 5% W/V of the lysine salt of bendazac were administered to 4 patients. Eye drops were instilled 3 times daily for one month. Even in this group of patients a clear-cut regression of the cataract was observed. In order to obtain the bendazac salt with a lysine the preparation is performed by heating,preferably in ethanol or water-acetone, equimolecular quantities of ^(1-benzyl-lH-indazol-3-yl)ox^[/ acetic acid and the aminoacid. The salt crystallizes on cooling as a bihydrate.

The so-obtained salt can b.e used as such or after drying under vacuum to constant weight. Both the bihydrate and the anhydrous salts with one of two optically active forms of lysine, as well, as the salt with racemic lysine can be employed in different pharmaceutical forms.

In use, the compound of the invention is administered orally in conventional formulations, namely in association with pharmaceutical excipients generally used for the production of compositions for oral administration,'"Singles doses between 0.3 g and 1 g have to be administered 2-3 times a day. administration may be used such as tablets and capsules; the unit dose for both tablet and capsule of active ingredient may be 500 mg. sitions are the excipients known in the pharmacist art. In the preparation of tablets, typical excipients include disintegrating agents, e. g. maize starch and lubricant agents-, e. g. magnesium stearate; in the preparation of capsules, standard gelatin capsules may be used containing the active ingredient alone or admixed Conventional pharmaceutical compositions for oral The carriers used in the preparation of these compo- with a diluent. »1> 199546 \ Typical and actual formulations are shown below Tablets Each tablet contains: Bendazac L. lysine salt hydrate mg 500. 0 cellulose ti 91. maize starch 11 75. 0 formalin-casein ti . 0 polyvinyl pyrrolidone ii . 0 silicon dioxide 11 7. magnesium stearate II 6. 0 hydroxy propyl cellulose 11 3. 0 talc 1! 1.

Hard gelatine capsules Each capsule contains: Bendazac L.lysine salt Maize starch lactose magnesium stearate hydrate mS 500.0 " 75.0 75.0 " 4.5 Granules Each simple doie (in paper envelope) contains: bendazac L. lysine salt hydrate g 0. 5 saccarose " 4,85 " ^ orange flavour "0,15 •=* o> 130546 Syrup 100 ml contain: . bendazac L.lysine salt g 5.0 hydrate saccarose "oo.O ethyl p-hydroxybenzoate " 0.15 sodium benzoate " 0. 5 raspberry flavour " 0.5 distilled water to 100 ml 1% eye drops 100 ml contain: bendazac L.lysine salt hydrate g 1.00 hydroxy propyl-methyl-cellulose g 0.500 H NaPO . H O g 0.018 z 4. z * HNa P0 . 12H 0 g CX<190 2 4^ NaCl g 0.680 thimerosal -g 0.001 water for injections to 100 ml 0. 5% eye drops bendazac L.lysine salt hydrate g 0. 50 hydroxy-propyl-methyl cellulose g 0. 500 H Na P0 .HO g 0.018 z 4 2 g 0.190 HNa P0 .12H 0 e 2 4 2 NaCl g 0. 740 thimerosal g 0.001 water for injections to 100 ml . ^M/ft

Claims (12)

- 15- Th e following non-restrictive examples illustrate the preparation of a salt which is the matter of the present invention. Example 1 Salt of /_{l-benzyl-lH-indazol-3-yl)ox^/ acetic acid with L. lysine. 12 g (0.042 mol) of ^(l-benzyl_lH-indazol-3-yl) ox%/ acetic acid and 6.2 g (0.042 mol) of L. lysine were dissolved in 100 ml of 95% ethanol by heating. The mixture was filtered warm in order to remove the very small amounts of impurities and it was then left to stand overnight at room temperature. The crystalline product was filtered and recrystallized from 9 5% ethanol: 15 g of the salt were first obtained (yield 75»9%) 'while the mother-liquor yielded more product. The,compound consists of one molecule of ^(l-benzyl-lH-indazol-3-yl)ox^/ acetic acid, one of L. lysine and two of crystallization-water; the same shows melting point 178-81°C with loss of water during heating. Example 2 The salt obtained according to the Example 1 was dried under vacuum (5-10 Torr.) at 105°C to constant weight: in such a form it shows melting point 17 8-8l°C with decomposition. ^3/^vv 199546 - 16 - What we claim is;

1. A salt of [(l-benzyl-lH-indazol-3-yl)oxy ] acetic acid with lysine.

2. The salt of I(l-benzyl-lH-indazol-3-yl)oxy] acetic acid with L.lysine.

3. The salt according to claim 2 which contains crystallization water.

4. The salt according to claim 2 which is anhydrous.

5. A process for preparing a salt of claim 1 or claim 2 consisting in that equimolecular quantities of [(1-benzyl-lH-indazol-3-yl)oxy] acetic acid and a lysine are dissolved by heating in a solvent, from which the salt crystallizes on cooling and is then separated and obtained in hydrate form.

6. A process according to claim 5 in which the said lysine salt is separated and obtained in an anhydrous form.

7. A therapeutic composition useful in the treatment of cataract, in an acceptable pharmaceutical carrier and containing a bendazac lysine salt in a form suitable to obtain effective tissue concentrations.

8. A composition according to claim 7 in the form of eye drops containing 0.25% W/V of a lysine salt of bendazac.

9. A composition according to claim 7 prepared for oral administration.

10. A method for the treatment of cataract in non-human mammals, consisting in the administration to patients with cataract of a bendazac lysine salt in an acceptable pharmaceutical carrier and in 17 199546

11. A method according to claim 10 in which a lysine salt of bendazac is administered by. oral route at a daily dose of 300-1000 mg optionally divided in two or more daily administrations.

12. A lysine salt of bendazac substantially as specifically described herein in either of the Examples. BALDWIN SON & CAREY Attorneys for the applicants. 23 jygg