WO2008061647A1

WO2008061647A1 - Use of a compound as vegf inhibitor

Info

Publication number: WO2008061647A1
Application number: PCT/EP2007/009767
Authority: WO
Inventors: Torbjörn Lundstedt; Gunilla EKSTRÖM
Original assignee: Acure Pharma Ab
Priority date: 2006-11-23
Filing date: 2007-11-12
Publication date: 2008-05-29
Also published as: WO2008061647A8; GB0623381D0

Abstract

There is disclosed the use of N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof as an agent for use in the inhibition of Vascular Endothelial Growth Factor (VEGF). Such an agent is likely to be of application in the treatment or prevention of undesired blood vessel formation during tumour growth and/or in inflammatory conditions.

Description

Use of a compound as VEGF inhibitor.

The present invention relates to the use of N-(2-chloro-3,4-dimethoxybenzylidene- amino)-guanidine for the treatment, hindrance or prevention of blood vessel development and formation (angiogenesis).

A number of benzylideneamino guanidines known in the art have shown anti- depressive effects (US 4060640). Examples of other pharmacologically active guanidines known in the art are described in patents US3982020, GBl 223491, WO02/11715, WO02/81430 and WO02/80896. Other application areas are also known in the art and are described in patents US3896332, DEl 165013, and US3941825. Even though the benzylideneamino guanidines known from the prior art are structurally similar to the compound in the present invention, they have never been reported as, and there is no suggestion that they might be, effective to inhibit vascular endothelial growth factor (VEGF).

Vascular Endothelial Growth Factor (VEGF) is a mitogen primarily for vascular endothelial cells and is structurally related to platelet-derived growth factor. It is a stimulator of endothelial cell proliferation that has been implicated in the vascularisation and growth of a variety of tumours, and is believed a major regulator of tumour angiogenesis in vivo. It is also implicated in some inflammatory conditions. For the treatment (reduction, prevention or hindrance) of undesired blood vessel formation during tumour growth and/or in inflammatory conditions, a VEGF inhibitor is likely to play a useful role.

We have now surprisingly found that N-(2-chloro-3,4-dimethoxybenzylideneamino)- guanidine and salts thereof can act as a VEGF inhibitor. Accordingly, in one aspect, the invention provides N-(2-chloro-3,4 dimethoxybenzylideneamino)-guanidine and salts thereof for use as a VEGF inhibitor. The invention further provides the use of N-(2-chloro-3,4-dimethoxybenzylidene- amino)-guanidine in the manufacture of a medicament for use as a VEGF inhibitor.

The invention also relates to methods for the manufacture of and pharmaceutical preparations comprising N-(2-chloro-3,4-dimethoxybenzylideneamino)-guanidine, and salts thereof, as well as to their use in medical and veterinary practice related to inhibition of vascular endothelial growth factor (VEGF). Such practice may include, but is not limited to, the inhibition or prevention of blood vessel formation and the treatment of angiogenesis.

EXAMPLES

The following examples are intended to illustrate but not to limit the scope of the invention.

Example 1

Preparation of N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine A solution of 2-chloro-3,4-dimethoxybenzaldehyde (1.0 g, 5 mmol), aminoguanidine bicarbonate (0.68 g, 5 mmol) and acetic acid (1 ml), in 15 ml of methanol was heated at reflux for 10 min. The reaction mixture was cooled down to 0 ⁰C and the residue was filtered off.. The filtrate was evaporated under vacuum and the product was crystallised from ethanol. Yield of the title compound was 1.1 g (70%), M.p. 198-200 ⁰C. Example 2

This example illustrates the potency of N-(2-chloro-3,4-dimethoxybenzylidene- amino)-guanidine and its therapeutically active acid addition salts for treatment of blood vessel formation.

Method

In rats with induced arthritis, antigen induced arthritis (AIA), pannus tissue from knee- joints was taken on day four after challenge. (Andersson, S. E., Lexmuller, K., and Ekstrom, G. M. 1998. Physiological characterization of mBSA antigen induced arthritis in the rat. I. Vascular leakiness and pannus growth. J Rheumatol 25:1772-1777).

The tissue was digested with collagenase type I, and the cells were allowed to proliferate for 7-10 days before stimulation and exposure to the compounds. The culture consisted of. a mixed cell population with the principal constituents being fibroblasts and macrophages (Andersson, S. E., Johansson, L. H., Lexmuller, K., and Ekstrom, G. M. 2000. Anti-arthritic effect of methotrexate: is it really mediated by adenosine? Eur J Pharm Sci 9:333-343).

The cells were stimulated with IL-I with simultaneous addition of AMAP 102. After three days of stimulation and compound exposure, the supernatants were collected, and the VEGF content measured using ELISA (DuoSet^® rat VEGF, R&D Systems, MN, USA). The cell viability was taken into account using the Cell Proliferation Reagent WST-I.

The extent to which N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine reduced the ILl -induced VEGF production in primary rat synoviocytes in a dose-dependent manner in the 25-100 μM range is shown in Fig. l hereto.

Suitable forms of pharmaceutical preparation for administration according to the invention include for example tablets, capsules, solutions, syrups, or emulsions. The content of the pharmaceutically effective compound should desirably be in the range from 0.1 to 5 wt. % , of the total composition. The preparations may be administered orally in the form of a tablet, as a powder, as a powder in a capsule (e.g. a hard gelatine capsule), as a solution or suspension.

It is preferable if the composition is administered orally. Suitable tablets may be obtained, for example, by mixing the active substance(s) with known carriers, diluents or excipients, such as calcium carbonate, calcium phosphate or lactose, disintegrants such as corn starch or alginic acid, binders such as starch or gelatine, lubricants such as magnesium stearate or talc and/or agents for delaying release, such as carboxymethyl cellulose, cellulose acetate phthalate, or polyvinyl acetate. The tablets may also comprise several layers.

Coated tablets may suitably be prepared by coating cores produced similarly to the tablets with substances normally used for tablet coatings, for example collidone or shellac, gum arabic, talc, titanium dioxide or sugar. The tablet coating may consist of a number of layers to achieve delayed release, possibly using the excipients mentioned above for the tablets.

Syrups containing the active substances or combinations thereof according to the invention may additionally contain a sweetener such as saccharin, cyclamate, glycerol or sugar and a flavour enhancer, e.g. a flavouring such as vanillin or orange extract. They may also contain suspension adjuvants or thickeners such as sodium carboxymethyl cellulose, wetting agents such as, for example, condensation products of fatty alcohols with ethylene oxide, or preservatives such as /7-hydroxybenzoates.

Capsules containing one the active substance may for example be prepared by mixing the active substance with inert carriers such as lactose or sorbitol and packing them into gelatine capsules. Suppositories may be made for example by mixing with carriers provided for this purpose, such as neutral fats or polyethyleneglycol or the derivatives thereof. Excipients which may be used include, for example, water, pharmaceutically acceptable organic solvents such as paraffins (e.g. petroleum fractions), vegetable oils (e.g. groundnut or sesame oil), mono- or polyfunctional alcohols (e.g. ethanol or glycerol), carriers such as e.g. natural mineral powders (e.g. kaolins, clays, talc, chalk), synthetic mineral powders (e.g. highly dispersed silicic acid and silicates), sugars (e.g. cane sugar, lactose and glucose), emulsifiers (e.g. lignin, spent sulphite liquors, methylcellulose, starch and polyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc, stearic acid and sodium lauryl sulphate).

For oral administration the tablets may contain, in addition to the abovementioned carriers, additives such as sodium citrate, calcium carbonate and dicalcium phosphate together with various additives such as starch, preferably potato starch, gelatine and the like. Moreover, lubricants such as magnesium stearate, sodium lauryl sulphate and talc may be used at the same time for the tabletting process. In the case of aqueous suspensions, the active substances may be combined with various flavour enhancers or colourings in addition to the excipients mentioned above.

A solution for parenteral administration by injection can be prepared in an aqueous solution of a water-soluble pharmaceutically acceptable acid addition salt of the active substance preferably in a concentration of 0.1 % to about 5 % by weight. These solutions may also contain stabilising agents and/or buffering agents.

EXAMPLE 3

Example of a preparation comprising a capsule

Per capsule Active ingredient, as salt 5 mg

Lactose 250 mg

Starch 120 mg Magnesium stearate 5 mg

Total up to 380 mg In case higher amounts of active ingredient, the amount of lactose used may be reduced. The ingredients are then packed into a gelatin capsule.

Example of a suitable tablet formulation.

Per tablet

Active ingredient, as salt 5 mg

Potato starch 238 mg

Colloidal Silica 10 mg

Talc 20 mg

Magnesium stearate 2 mg

5 % aqueous solution of gelatine 25 me

Total up to 300 mg

Claims

Claims:

1. N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof for use as a VEGF inhibitor.

2. N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof for use to inhibit or prevent blood vessel formation.

3. N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof for the treatment of angiogenesis.

4. The use of N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof in the preparation of a medicament for use as a VEGF inhibitor.

5. The use of N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof in the preparation of a medicament for use to inhibit or prevent blood vessel formation.

6. The use of N-(2-chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof in the preparation of a medicament for the treatment of angiogenesis.

7. A method of inhibiting the action of VEGF in a subject which comprises administering to said subject an effective amount of N-(2-chloro-3,4- dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof.

8. A method of inhibiting or preventing blood vessel formation in a subject which comprises administering to said subject an effective amount of N-(2- chloro-3,4-dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof.

9. A method of treating angiogenesis in a subject which comprises administering to said subject an effective amount of N-(2-chloro-3,4- dimethoxybenzylideneamino)guanidine or a pharmacologically acceptable salt thereof.