HK1101065B - Use of substituted 1,2,3 indolizine derivatives for the preparation of medicaments for treating diseases associated with a pathological choroidal angiogenesis - Google Patents

Description

Use of 1,2, 3-substituted indolizine derivatives for the preparation of a medicament for the treatment of pathologies associated with choroidal angiogenesis

It has been shown in vitro and in vivo that several growth factors are involved in producing a "pro-angiogenic" imbalance, which leads to the uncontrolled proliferation of endothelial cells observed in the neovascular phenomenon. FGF2 or bFGF (fibroblast growth factor 2 or b) was first and best characterized. FGF2 is a 18000D protein that induces the proliferation, migration, and production of proteases from cultured endothelial cells and neovascularization in vivo. FGF2 interacts with endothelial cells via two classes of receptors, the high affinity tyrosine kinase receptor (FGFR) and the low affinity heparan sulfate proteoglycan receptor (HSPG), located on the cell surface and in the extracellular matrix. FGF2 and its receptor therefore represent a very suitable therapeutic target for the purpose of inhibiting angiogenic processes (Keshet E., Ben-Sasson S.A., J.Clin.Invest., (1999), Vol.501, pp.104-1497; Presta M., Rustina M., Dell' Era P., Tanghetti E., Urbinati C., Giulini R. et al., New York: PleniumPublisers, (2000), pp.7-34, Billottet C., Janji B., Thiery J.P., Joanneauj., Oncogene, (2002), Vol.21, pp.8128-).

Recent studies have demonstrated the effect of FGF on neovascularization by experimental study models on the mouse eye. (Rousseau et al.

《Involvement of Fibroblasts Growth Factors in angiogenesis and retinervationization, Experimental Eye Research. Laboratoire des M ü ssismeMol Molluires de I' angiogen de INSERM EML01-13, Universal de Bordeax, France, 13 mai 2003, p.147-156). However, the precise role of FGFs, and the essence of their receptors in the process of ocular pathological angiogenesisThe exact action and in particular the precise action in the choroid canal is still poorly understood and sometimes results in a different approach than that assumed.

(Tobe et al. & gt Targeted delivery of the FGF2 gene dots not presentneovacuolization in a music model, am.J.Pathol.153, 1641-1646. 1998/Yamada et al, Cell in underlying unmask a prosthetic nutritional type microorganism with an embedded expression of FGF2 in the retina, J.cell Physiol.185, 135-142, 2000).

International patent application WO03/084956 discloses FGF inhibitory derivatives, in particular for the treatment of pathologies in which angiogenesis appears to have a significant effect on the progression of, for example, chronic inflammatory diseases such as rheumatoid arthritis or IBD (inflammatory bowel disease). International patent application WO03/084956 also discloses that the compounds are useful in the treatment of diseases caused by diabetic vascular complications, such as diabetic retinopathy, in which retinal blood vessels are ruptured or blocked.

It has now been found that certain 1,2, 3-substituted indolizine (indolizine) derivatives, which are antagonists of FGF receptors known as FGFRs, are useful in the treatment of diseases associated with pathological choroidal angiogenesis. Surprisingly, these compounds are active when administered orally, topically, and especially intraocularly. The invention thus relates to the use of a 1,2, 3-substituted indolizine derivative, corresponding to the following general formula I, optionally in the form of one of its pharmaceutically acceptable salts, for the preparation of a medicament useful for the treatment of pathologies associated with choroidal angiogenesis:

wherein R is₁Represents hydroxyl, straight chain or branched chain of 1-5 carbon atomsAlkoxy, carboxyl, alkoxycarbonyl of 2 to 6 carbon atoms, or a group of the formula:

·- NR5R6

·- NH-SO2-Alk

·- NH-SO2-Ph

·- NH-CO-Ph

·-N(Alk)-CO-Ph

·-NH-CO-NH-Ph

·-NH-CO-Alk

·-NH-CO₂-Alk

·-O-(CH₂)_n-cAlk

·-O-Alk-COOR₇

·-O-Alk-O-R₈

·-O-AlK-OH

·-O-Alk-C(NH₂):NOH

·-O-Alk-NR₅R₆

·-O-Alk-CN

·-O-(CH₂)_n-Ph

·-O-Alk-CO-NR₅R₆

·-CO-NH-(CH₂)_m-COOR₇

·-CO-NH-Alk

wherein:

alk represents a linear or branched alkyl or alkylene group having from 1 to 5 carbon atoms,

cAlk represents a cycloalkyl group of 3 to 6 carbon atoms,

n represents an integer from 0 to 5,

m represents an integer from 1 to 5,

·R₅and R₆Identical or different, represent a hydrogen atom, a linear or branched alkyl group of 1 to 5 carbon atoms, or a benzyl group,

·R₇represents a hydrogen atom, or an alkyl group of 1 to 5 carbon atoms,

·R₈table 1-5 alkyl groups of carbon atoms or-CO-Alk groups,

ph represents a phenyl group, optionally substituted by one or more halogen atoms, by one or more alkoxy groups of 1 to 5 carbon atoms, by one or more carboxyl groups, or by one or more alkoxycarbonyl groups of 2 to 6 carbon atoms,

R₂represents a hydrogen atom, an alkyl group of 1 to 5 carbon atoms, an alkylhalo group of 1 to 5 carbon atoms containing 3 to 5 halogen atoms, a cycloalkyl group of 3 to 6 carbon atoms or a phenyl group optionally substituted by one or more halogen atoms, by one or more alkoxy groups of 1 to 5 carbon atoms, by one or more carboxyl groups, or by one or more alkoxycarbonyl groups of 2 to 6 carbon atoms,

a represents-CO-, -SO-or-SO₂-a group of,

R₃and R₄Which are identical or different, represent a hydrogen atom, an alkoxy group having 1 to 5 carbon atoms, an amino group, a carboxyl group, an alkoxycarbonyl group having 2 to 6 carbon atoms, a hydroxyl group, a nitro group, a hydroxyamino group, a group of the formula:

·-Alk-COOR₇

·-NR₅R₆

·-NH-Alk-COOR₇

·-NH-COO-Alk

·-N(R₁₁)-SO₂-Alk-NR₉R₁₀

·-N(R₁₁)-SO₂-Alk

·-N(R₁₁)-Alk-NR₅R₆

·-N(R₁₁)-CO-Alk-NR₉R₁₀

·-N(R₁₁)-CO-Alk

·-N(R₁₁)-CO-CF₃

·-NH-Alk-HetN

·-O-Alk-NR₉R₁₀

·-O-Alk-CO-NR₅R₆

·-O-Alk-HetN

wherein n, m, Alk, R₅、R₆And R₇Has the above structure of R₁The meanings given; and is

R₉And R₁₀Which are identical or different and each represent a hydrogen atom or an alkyl group having from 1 to 5 carbon atoms,

R₁₁represents a hydrogen atom or-Alk-COOR₁₂Group, wherein R₁₂Represents a hydrogen atom, an alkyl group of 1 to 5 carbon atoms or a benzyl group,

HetN represents a 5-or 6-membered heterocyclic ring containing at least one nitrogen atom and optionally another heteroatom selected from nitrogen and oxygen;

or R₃And R₄Together form an unsaturated 5-or 6-membered heterocyclic ring, provided however that when R₃Represents alkoxy and R₄represents-O-Alk-NR₉R₁₀When radical or hydroxy, R₁Alkoxy groups are not represented.

Preference is given to using compounds of the general formula I, optionally in the form of one of its pharmaceutically acceptable salts:

wherein R is₁Represents a hydroxyl group, a linear or branched alkoxy group of 1 to 5 carbon atoms, a carboxyl group, an alkoxycarbonyl group of 2 to 6 carbon atoms, or a group of the formula:

·-NR₅R₆

·-NH-SO₂-Alk

·-NH-SO₂-Ph

·-NH-CO-Ph

·-N(Alk)-CO-Ph

·-NH-CO-NH-Ph

·-NH-CO-Alk

·-NH-CO₂-Alk

·-O-(CH₂)_n-cAlk

·-O-Alk-COOR₇

·-O-Alk-O-R₈

·-O-Alk-OH

·-O-Alk-NR₆R₈

·-O-Alk-CN

·-O-(CH₂)_n-Rh

·-O-Alk-CO-NR₅R₆

·-CO-NH-(CH₂)_m-COOR₇

·-CO-NH-Alk

wherein:

alk represents a linear or branched alkyl or alkylene group having from 1 to 5 carbon atoms,

cAlk represents a cycloalkyl group of 3 to 6 carbon atoms,

n represents an integer from 0 to 5,

m represents an integer from 1 to 5,

·R₅and R₆Identical or different and each represents a hydrogen atom, a linear or branched alkyl group of 1 to 5 carbon atoms or a benzyl group,

·R₇represents a hydrogen atom, or an alkyl group of 1 to 5 carbon atoms,

·R₈represents an alkyl group of 1 to 5 carbon atoms or a-CO-Alk group,

ph represents a phenyl group, optionally substituted by one or more halogen atoms, by one or more alkoxy groups of 1 to 5 carbon atoms, by one or more carboxyl groups, or by one or more alkoxycarbonyl groups of 2 to 6 carbon atoms,

R₂represents alkyl of 1 to 5 carbon atoms, trifluoromethyl, cycloalkyl of 3 to 6 carbon atoms or phenyl, which phenyl is optionally substituted by one or more halogen atoms, by one or more alkoxy of 1 to 5 carbon atoms, by one or more carboxyl groups, or by one or more alkoxycarbonyl of 2 to 6 carbon atoms,

a represents-CO-or-SO₂-a group of,

R₃and R₄Which are identical or different, represent a hydrogen atom, an alkoxy group having 1 to 5 carbon atoms, an amino group, a carboxyl group, an alkoxycarbonyl group having 2 to 6 carbon atoms, a nitro group, a hydroxyamino group, a group of the formula:

·-Alk-COOR₇

·-NR₅R₆

·-NH-Alk-COOR₇

·-NH-COO-Alk

·-N(R₁₁)-SO₂-Alk-NR₉R₁₀

·-N(R₁₁)-SO₂-Alk

·-N(R₁₁)-Alk-NR₅R₆

·-N(R₁₁)-CO-Alk-NR₉R₁₀

·-N(R₁₁)-CO-Alk

·-N(R₁₁)-CO-CF₃

·-NH-Alk-HetN

wherein n, m, Alk, R₅、R₆And R₇Has the above structure of R₁The meanings given; and is

R₉And R₁₀Which are identical or different, represent a hydrogen atom or an alkyl group having from 1 to 5 carbon atoms,

R₁₁represents a hydrogen atom or-Alk-COOR₁₂Group, wherein R₁₂Represents a hydrogen atom, an alkyl group of 1 to 5 carbon atoms or a benzyl group,

HetN represents a 5-or 6-membered heterocyclic ring containing at least one nitrogen atom and optionally another heteroatom selected from nitrogen and oxygen.

Particular preference is given to the use of compounds of the formula I, optionally in the form of one of its pharmaceutically acceptable salts, wherein

R₁Represents an alkoxy group of 1 to 5 carbon atoms, a carboxyl group, an-O-Alk-COOH group, wherein Alk represents a linear or branched alkylene group of 1 to 5 carbon atoms, a group of formula-O-Alk-Ph, wherein Alk represents an alkylene group of 1 to 5 carbon atoms and Ph represents a phenyl group, which may optionally be substituted by one or more halogen atoms or one or more alkoxy groups of 1 to 5 carbon atoms or one or more carboxyl groups, a group of formula-NH-CO-Ph, a group of formula-NH-SO₂A group of-Ph, or a group of formula-NH-CO-NH-Ph,

R₂represents an alkyl group of 1 to 5 carbon atoms,

a represents a-CO-group,

R₃and R₄Which, in each case, represent a hydrogen atom, an alkoxy group having 1 to 5 carbon atoms, an amino group, a carboxyl group, an alkoxycarbonyl group having 2 to 6 carbon atoms.

Particular preference is given to using a compound selected from the following formula I, optionally in the form of one of its pharmaceutically acceptable salts:

(4-amino-3-methoxyphenyl) (1-methoxy-2-methylindolizin-3-yl) methanone

3- (4-amino-3-methoxybenzoyl) -2-methylindolizin-1-yl carboxylic acid

2- { [3- (4-amino-3-methoxybenzoyl) -2-methylindolizin-1-yl ] oxy } acetic acid

(4-amino-3-methoxyphenyl) {1- [ 4-chlorobenzyl) oxy ] -2-methylindolizin-3-yl } methanone

(4-amino-3-methoxyphenyl) {1- [ (3-methoxybenzyl) oxy ] -2-methylindolizin-3-yl } methanone

4- ({ [3- (4-amino-3-methoxybenzoyl) -2-methylindolizin-1-yl ] oxy } methyl) benzoic acid

3- (4-Carboxybenzoyl) -2-methylindolizin-1-yl carboxylic acid

3- [ (1-methoxy-2-methylindolizin-3-ylcarbonyl ] benzoic acid methyl ester

4- [ (1-methoxy-2-methylindolizin-3-yl) carbonyl ] benzoic acid

2-amino-5- [ (1-methoxy-2-methylindolizin-3-yl) carbonyl ] benzoic acid

2-amino-5- ({1- [ (3-methoxybenzoyl) amino ] -2-methylindolizin-3-yl } carbonyl) benzoic acid

2-amino-5- ({ 2-methyl-1- [ (3, 4, 5-trimethoxybenzoyl) amino ] indolizin-3-yl } carbonyl) benzoic acid

2-amino-5- ({1- { [ (3-methoxyphenyl) sulfonyl ] amino) -2-methylindolizin-3-yl } carbonyl) benzoic acid.

Pathological choroidal angiogenesis is a process that produces new capillary blood vessels of the choroid. The choroid is the highly vascularized membrane of the eye: an intact network of fine capillaries, which supply nutrition to the iris and retina at their distal ends. Pathological choroidal angiogenesis is primarily due to the appearance of an "preangiogenic" imbalance, but also due to deleterious changes in Bruch's membrane located beneath the retina. Thus, choroidal capillaries proliferate uncontrollably and invade the subretinal space via defects in Bruch's membrane (lafout et al, br.j. ophtalmol., 84, 239-.

Among the diseases associated with pathological choroidal angiogenesis, in particular due to abnormalities in the Bruch membrane, age-related macular Degeneration (DMLA), high myopia (quanta et al, Graefe's arch, clin. exp. ophtalmol., 238, 101-.

Thus, according to one aspect of the present invention, the present invention relates to the use of a 1,2, 3-substituted indolizine derivative of general formula I in the manufacture of a medicament useful in the treatment of a disease associated with pathological choroidal angiogenesis, such as age-related macular Degeneration (DMLA), high myopia, pseudoxanthoma, putative histoplasmosis syndrome, toxoplasmosis, sarcoidosis, or behcet's disease.

According to another aspect of the invention, a subject of the present invention is a pharmaceutical composition comprising at least one active ingredient corresponding to a compound of formula I or one of its pharmaceutically acceptable salts, optionally in combination with one or more inert and suitable excipients, for the treatment of diseases associated with pathological choroidal angiogenesis.

In particular, a subject of the present invention is a pharmaceutical composition comprising at least one active ingredient corresponding to a compound of formula I, optionally in combination with one or more inert and suitable excipients, selected from:

(4-amino-3-methoxyphenyl) (1-methoxy-2-methylindolizin-3-yl) methanone

3- (4-amino-3-methoxybenzoyl) -2-methylindolizin-1-yl carboxylic acid

2- { [3- (4-amino-3-methoxybenzoyl) -2-methylindolizin-1-yl ] oxy } acetic acid

(4-amino-3-methoxyphenyl) { l- [ 4-chlorobenzyl) oxy ] -2-methylindolizin-3-yl } methanone

(4-amino-3-methoxyphenyl) {1- [ (3-methoxybenzyl) oxy ] -2-methylindolizin-3-yl } methanone

4- ({ [3- (4-amino-3-methoxybenzoyl) -2-methylindolizin-l-yl ] oxy } methyl) benzoic acid

3- (4-Carboxybenzoyl) -2-methylindolizin-l-ylcarboxylic acid

3- [ (1-methoxy-2-methylindolizin-3-ylcarbonyl ] benzoic acid methyl ester

4- [ (l-methoxy-2-methylindolizin-3-yl) carbonyl ] benzoic acid

2-amino-5- [ (1-methoxy-2-methylindolizin-3-yl) carbonyl ] benzoic acid

2-amino-5- ({1- [ (3-methoxybenzoyl) amino ] -2-methylindolizin-3-yl } carbonyl) benzoic acid

2-amino-5- ({ 2-methyl-l- [ (3, 4, 5-trimethoxybenzoyl) amino ] indolizin-3-yl } carbonyl) benzoic acid

2-amino-5- ({1- { [ (3-methoxyphenyl) sulfonyl ] amino) -2-methylindolizin-3-yl } carbonyl) benzoic acid.

The excipients are selected according to the pharmaceutical dosage form and the desired method of administration (oral, intraocular or topical).

The pharmaceutical compositions of the present invention are preferably administered orally or intraocularly.

Particularly preferably, the pharmaceutical composition of the present invention is administered orally.

In the pharmaceutical compositions of the invention for oral administration, the active ingredient may be administered in unit dosage form, as a mixture with conventional pharmaceutical carriers. Suitable unit forms of administration to be taken orally include, for example, tablets (optionally divisible), capsules, powders, granules and solutions or suspensions.

When preparing solid compositions in the form of tablets, the principal active ingredient is mixed with a pharmaceutical vehicle such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic, and the like.

The tablets may be coated with sucrose or other suitable material, or they may be treated to provide them with prolonged or delayed activity and sustained release of a predetermined amount of the active ingredient.

Formulations in the form of gelatin capsules may be obtained by mixing the active ingredient with a diluent and filling the resulting mixture into soft or hard gelatin capsules.

Formulations in the form of syrups or elixirs may include mixing the active ingredient with sweetening agents, preferably non-caloric sweetening and methylparaben and propylparaben as preservatives, suitable coloring and flavoring agents.

The water-dispersible powders or granules may contain the active ingredient in admixture with dispersing agents, wetting agents, or suspending agents such as polyvinylpyrrolidone and sweetening or flavoring agents.

The active ingredient may also be formulated in the form of microcapsules, optionally with one or more carriers or additives.

In the pharmaceutical composition of the present invention, the active ingredient may also be in the form of an inclusion complex in a cyclodextrin, an ether thereof or an ester thereof.

The pharmaceutical composition for topical administration may be various ophthalmic formulations. They may comprise the active ingredient, together with ophthalmologically acceptable preservatives, surfactants, binders, agents promoting the penetration of the active ingredient, buffers, sodium chloride and water, in order to obtain sterile solutions or suspensions for ophthalmic use.

Ophthalmic solutions can be prepared by dissolving the active ingredient in an isotonic buffer solution. These solutions may include surfactants to aid in the dissolution of the active ingredient. For better application, the ophthalmic solution may also include a thickening agent such as hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, or polyvinylpyrrolidone.

To prepare sterile ophthalmic ointments, the active ingredient is combined with a preservative in a suitable vehicle such as liquid lanolin. Sterile gels can be prepared by suspending the active ingredient in a hydrophilic group comprising carbopol 940 or other similar compound.

For topical application, the pharmaceutical composition according to the invention is preferably in the form of a solution or suspension at pH 4-8. The concentration of the active ingredient is between 0.0001% and 5% w/w, preferably between 0.0001% and 1%. Depending on the order of the physician, these compositions can be administered in 1-2 drops, 1-4 times daily.

The amount of active ingredient to be administered depends, as usual, on the degree of progression of the disease and on the age and weight of the patient.

Specifically, the orally administrable pharmaceutical compositions of the invention contain a recommended dose of the active ingredient of 1-900 mg/kg/day.

Preferably, the orally administrable pharmaceutical compositions of the invention contain a recommended dose of active ingredient of 3-300 mg/kg/day.

Particularly preferably, the orally administrable pharmaceutical compositions of the invention contain a recommended dose of the active ingredient of 1 to 100 mg/kg/day.

The following examples illustrate the invention without limiting it.

Example (b): laser induction of choroidal neovascularization

1. Apparatus and method

C57BI6 mice were used in these experiments. A laser-induced choroidal neovascularization model (4 strokes per eye in the vicinity of the papilla) was applied to the animals as described by Tobe et al in am.j.pathol., 1998.

After induction and 14 days after detection by angiography (which allows assessment of the percentage of damage developing neovascularization), animals were sacrificed and the eyeballs removed for histological analysis. During the last 7 days, mice were orally administered one of the compounds of formula I. For the monohydrate of the sodium salt of 2-amino-5- [ (1-methoxy-2-methylindolizidin-3-yl) carbonyl ] benzoic acid, the dose chosen was 30 mg/kg/day. The vehicle used was a 0.6% solution of methylcellulose in water.

4 mice were used as a control group and 6 mice were used as a treatment group.

Morphological assessment of thickness was performed using a computerized image analysis system, and dimensions of neovascular responses were estimated from frozen sections. These sections were either simply stained with hematoxylin or examined immunofluorescently using an antibody (anti-CD 31) that localizes the vascular structure.

Evaluation was performed by measuring the ratio B/C between EPR and the peak of neovascular response ("B") on the one hand, and the adjacent intact choroid layer thickness ("C") on the other hand. Such a quantification system is preferred to measure wound surface area because it is independent of the positioning of the tissue slices.

2. Results

Angiography performed at day 14 showed induction of new blood vessels by laser in 72% of wounds in control mice.

Histological analysis and immunohistochemistry using the antibody anti-CD 31 confirmed the presence of newly formed capillaries in these areas, which showed significant spreading of fluorescein in the angiograms in the control group. In contrast, animals treated with the compound showed only moderate thickening where the shock occurred, with no obvious signs of neovascularization.

Quantification of the laser-induced response by measuring the B/C ratio showed a significant reduction of the response in mice treated with the compounds of the invention, compared to control mice, of the order of 50% (p < 0.001).

These results confirm the great advantage of the use of the compounds of formula I for the preparation of a medicament useful for the treatment of pathological choroidal angiogenesis-related diseases.

Claims (2)

1. Use of 2-amino-5- [ (1-methoxy-2-methylindolizidin-3-yl) carbonyl ] benzoic acid or a sodium salt thereof in the manufacture of a medicament for inhibiting pathological choroidal angiogenesis.
2. 2. Use according to claim 1, for the preparation of a medicament for the treatment of DMLA or high myopia, age-related macular degeneration induced by pathological choroidal angiogenesis.