GB2262524A

GB2262524A - Pharmaceutically active 2-hydroxy-benzylamines

Info

Publication number: GB2262524A
Application number: GB9126831A
Authority: GB
Inventors: Helmut Egger; Peter Nussbaumer; Neil S Ryder
Original assignee: Sandoz AG
Current assignee: Sandoz AG
Priority date: 1991-12-18
Filing date: 1991-12-18
Publication date: 1993-06-23
Also published as: GB9126831D0

Abstract

Compounds of the formula <IMAGE> wherein R1, R2 and R3 are the same or different and represent hydrogen, hydroxy, trifluormethyl, alkyl, alkoxy, acyloxy, halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, and R4 represents hydrogen or a group of formula <IMAGE> wherein R5 represents hydrogen or alkyl and R6, R7 and R8 are the same or different and represent hydrogen, hydroxy, trifluormethyl, alkyl, alkoxy, acyloxy, halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, in free form or, where such forms exist, in acid addition salt form, are useful as PKC-inhibitors in the treatment of inflammatory diseases such as arthritis, in the treatment and prophylaxis of immunological disorders, of allergies, of akin diseases, in transplantations and in the field of oncology.

Description

Organic compounds, processes for their production and their use The present invention concerns new organic compounds, processes for their production, pharmaceutical compositions containing them and their use as pharmaceuticals especially as PKCinhibitors in the treatment of inflammatory diseases such as arthritis. in the treatment and prophylaxis of immunological disorders, of allergies, of skin diseases, in transplantations and in the field of oncology.

More particulariy the invention concerns compounds of the formula

wherein R1, R2 and R3 are the same or different and represent hydrogen, hydroxy, trifluormethyl. alkyl, alkoxy, acyloxy, halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, and R4 represents hydrogen or a group of formula

wherein RS represents hydrogen or alkyl and R6, R7 and R8 are the same or different and represent hydrogen, hydroxy, trifluormethyl, alkyl, alkoxy, acyloxy. halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, in free form or, where such forms exist. in acid addition salt form.

The present invention also provides a process for the production of compounds of formula which comprises reducing a compound of formula

wherein the substituents are as defined above, whereby in this reaction functional groups may be protected by suitable protecting groups. which may be removed subsequent to the reaction in conventional manner, and recovering the compounds thus obtained in free form or in acid addition salt form.

Functional groups may be protected by suitable protecting groups which may be removed subsequent to the reaction in conventional manner. End products can be isolated and purified according to known methods.

Unless otherwise stated alkyl moieties are preferably straight or branched chains having 1 to 12 expecially 1 to 8 carbon atoms. particuiarly 1 to 6 and especially 1 to 4. Any lower alkyl present as or in a substituent is straight or branched-chain and has preferably 1 to 4 especially 1 or 2 carbon atoms. R1, R2 and F3 represent preferably hydrogen or form together with the carbon atoms they are attached an aromatic ring, e.g. phenyl. Rs represents preferably hydrogen or methyl. Re. R7 and R8 represent preferably hydrogen or form together with the carbon atoms they are attached an aromatic ring, e.g. phenyl.

The process of the invention may be performed following standard procedures for the reduction of Schiff bases, preferably with a complex hydride such as NaBH4. It is preferably carried out in an inert solvent, e.g. in a lower alkanol such as methanol, and at room temperature.

The final products can be isolated and purified in conventional manner. Free bases of the compounds of formula I can be converted into salt forms and vice versa. Suitable acid addition salts are the hydrochloride, hydrogen fumarate or naphthalin-1 .5-disulphonate.

The starting compounds of formula Ill can be prepared by reaction of the corresponding amine of formula

with an aldehyde of formula

wherein the substituents are as defined above. This reaction may be carried out in conventional manner, e.g. in an inert solvent such as an aromatic hydrocarbon, e.g. toluene, and at room temperature or elevated temperature, preferably at lightly elevated temperature and in the presence of a water binding agent, e.g. molecuiar sieve. The thus obtained reaction products of formula Ill may be reacted further without isolation or purification to give the end products of formula I.

The amines of formula IV, wherein R4 represents a group of formula 11, can be prepared according to the following reaction scheme:

In this reaction scheme X represents a leaving group, e.g. halogen such as chlorine or a sulfonate, and the substituents are as defined above.

The remaining infermediates are either known or can be prepared analogously to known processes or e.g. as described in the examples.

The compounds of this invention possess advantageous ohemotherapeutical properties and exhibit on local. systemic or oral application a PKC-inhibitory activity. This activity can be shown in the following test: Protein kinase C inhibitory test: a) Preparation of enzyme Freshly removed rat brains are washed in ice-cold physiological saline and the remaining procedure carried out at 4 . Brains are homogenised in buffer H (12 ml per brain) in a Potter S glass homogeniser. The homogenate is spun at 12.OOOg 15 minutes. then at 1O0.OOOg 60 minutes. The supernatant is loaded onto a DEAE-sephacel column run on a FPLC system. For two brains, a 20 ml column equilibrated with buffer A is used.After washing with 12O ml buffer A (1 mllmin), a linear gradient from 0 to 100% buffer B (0.3 M NaCI in buffer A) is applied (100 ml, 1 ml/min) and collected in 2 ml fractions. Fractions containing PKC activity appear between 45 to 75% buffer B. Active fractions are desalted on PD-10 columns, pooled and stored in 1 ml aliquots containing 10% added glycerol in liquid nitrogen. Enzyme activity remains stable for at least two months.

Buffer H: Tris buffer pH 7,5 (20 mM); sucrose (250 mM); EGTA (10 mM); EDTA (2 mM); dithiothreitol (2 mM); PMSF (1 mM); leupeptin (10 mg/litre).

Buffer A: Tris buffer pH 7,5 (20 mM); EGTA (2 mM); EDTA (2 mM); dithiothreitol (2 mM); b) Enzyme assay Assay components Final cone Enzyme, diluted in 200 mM tris ph 7,5 25 ul/assay, protein conc. approx. 13 llg/ml phosphatidylserine 10 ,ug/ml 1-oieyl-2-acetyl-sn-glycerol 1 llg/ml histone IllS 200 Fg/ml calcium chloride 0,2 mM magnesium chloride 10 mM ATP 10 uM [P-32] ATP 1 ACi/ml distilled water, to a total assay volume of 250 ul.

The water-soluble components are used as 10-fold concentrated stock solutions. The lipids are prepared from chloroform stock solutions, dried under nitrogen and resuspended in water at 1 O-fold final concentration by sonication (1 ,5 min, microtip, in ice) to form a mixed emulsion (25111/assay). Enzyme, buffer, water, calcium and histone are pre-mixed as a cocktail. ATP and magnesium are pre-mixed as substrate cocktail (25 ul/assay). Test compounds are dissolved in dimethylsulphoxide at 100-fold concentration (2,5 ul/assay).

Assay sequence: 1. Enzyme cocktail 2. Lipids 3. Test compound 4. Pre-incubation 5 min 5. Substrate cocktail Assays are performed in plastic tubes incubated 5 min at 30 on a shaking waterbath. Assays are stopped by addition of 1 ml ice cold 25% trichloracetic acid pius 10 ul 2% serum albumin and placed in ice. Insoluble material is filtered on Whatman GFlC filter discs (2,5 cm), washed with 10% TCA, dried and counted for radioactivity.

Assays are performed with and without addition of lipids; PKC activity is defined as the difference between these two in terms of incorporation of P-32 into the TCA-insoluble fraction. All assays are in triplicate.

The compounds of the invention may therefore be used as PKC-inhibitors. For this use, the effective dosage will, of course, vary depending on the particular compound employed, the mode of administration and the treatment desired. However, in general, satisfactory results can be obtained when the compounds are administered at a daily dosage of from about 1 to 30 mglkg of animal body weight, given in suitably divided doses two to four times daily. For most large mammals, the total daily dosage is from about 70 to 2000 mg and dosage forms suitable for internal administration comprise about 17.5 to 1000 mg of the compound in admixture with a solid or liquid chemotherapeutical carrier or diluent. The compounds of the invention may be administered in similar manner to known standards for use in such indications.

The invention therefore also concerns a method of treatment of inflammatory diseases such as arthritis. of treatment and prophylaxis of immunological disorders, of allergies, of skin diseases, in transplantations and in the field of oncology, which comprises administering to a subject in need of such treatment an effective amount of a compound of the invention or a chemotherapeutically acceptable acid addition salt thereof as well as compounds of the invention or chemotherapeutically acceptable acid addition saits thereof for use as chemotherapeutic agents especially as PKC-inhibitory agents.

The compounds may be admixed with conventional chemotherapeutically acceptable diluents and carriers, and, optionally, other excipients and administered e.g. orally in such forms as tablets or capsules. The compounds may aiternatively be administered topically (in such conventional forms as ointments or creams), parenterally or intravenously. The concentrations of the active substance will, of course, vary depending on the compound employed. the treatment desired and the nature of the form etc. In general, however, satisfactory results are obtained e.g. in topical application forms at concentrations of from 0.05 to 5, in particular 0.1 to 1 wt.%. Such compositions also form part of the invention.

Preferred in the above mentioned indications is the compound of example 1, e.g. the N-me thyl-N- ( 1 -naphthylmethyl) -4- (2-hydroxybenzylamino) benzyl amine.

The invention also comprises the compounds of the invention for use as pharmaceuticals, especially for use as PKC-inhibitors.

The invention further comprises a method of treatment of comprising administering a therapeutically effective amount of a compound of the invention to a person in need of such a treatment.

In the following examples which illustrate the invention but in no way limit its scope references to temperature are in degrees celsius.

The following abbreviations are used: DEAE = diethylaminoethyl FPLC = fast protein liquid chromatography PKC = protein kinase C EGTA = ethylenebis (oxyethylenenitrilo) tetraacetic acid EDTA = ethylene diamino tetraacetic acid PMSF = phenylmethyl sulfonyl fluoride ATP = adenosinetriphosphate Example 1: N-Methyl-N- (1 -naphthylmethyl)-4- (2-hydroxyb6nzylamino) benzyl amine 0 2 g of molecular sieve (4 A) and 0,33 g of salicyl aldehyde are added to a solution of 0,75 g of N-methyl-N-(1-naphthylmethyl)-(4-amino)benzyl amine in 50 ml of toluene and the reaction mixture is stirred at 400 for 3 hours. Then the mixture is filtered, the toluene evaporated and the residue dissolved in 30 ml of methanol.After addition of 0,10 g of NaBH4 the reaction mixture is stirred at room temperature for 15 hours. The reaction mixture is acidified with 1N HCI, then neutralized with 1N NaOH, evaporated and the residue dissolved in ethyl acetate and water. The organic phases are washed with sodium chloride solution, dried over sodium sulfate and evaporated. Purification of the crude product with ether/petroleum benzine and chromatography (hexane/ethyl acetate = 2/1) gives the title compound as colourless crystals.

m.p.: 138-140 .

The following compounds are prepared analogously:

I Ex.: P1 R2 R3 R4 m.p.

Z 2 H H H -CH2. NH. CH2-0 glassy mass O 3 > H H beige, 120-1250 The required starting materials may be obtained as follows: A) N-Methyl-N-(1-naphthylmethyl)-(4-amino)benzyl amine a) N-Methyl-N-(1-naphthylmethyl)-(4-nitro3benzyl amine The mixture of 20 g of N-methyl-(1-aminomethyl)naphthaline.hydrochlorid, 16,9 g of 4-nitrobenzyl chloride and 20 g of powdered patassium carbonate in 200 ml of dry dimethylformamide is stirred at room temperature for 48 hours. Then the reaction mixture is poured onto ice water, extracted with ether, the organic phases are washed twice with water and once with saturated sodium chloride solution, dried over sodium sulfate and evaporated.Chromatography of the residue (toluenelethyl acetate = 10/1) gives the title substance as yellow oil.

b) N-Methyl-N-(I -naphthylmethyl)-(4-amino)benzyl amine 4,1 g of hydraziniumhydroxyd and 1,0 g of palladium black are added to a solution of 10,3 g of N-methyl-N-(1-naphthylmethyl)-(4-nitro)benzyl amine in 150 ml of ethanol and the mixture is refluxed for 2 hours. Then the catalyst is filtered off, and the solvent evaporated. Chro matography of the residue (toluene/ethyl acetate = 10/1) gives the title compound as colourless oil.

B) N- (1 -Naphthylmethyl)- (4-amino)benzyl amine Proceeding analogously as described under A) gives the title substance as colourless oil.

1K-NMR-Spectra Ex. Spectra: 11) 2.18 (s, 3H); 3.53 (s, 2H); 3.92 (s, 2H); 4.40 (s, 2H); 6.77-6.90 (m, 4H); 7.14-7.24 (m, 4H); 7.37-7.52 (m, 4H); 7.75-7.86 (m, 2H); 8.23 (m, 1H).

22) 3.62 (s, 2H); 4.08 (s, 2H); 4.18 (d, J=6Hz, 2H) 5.89 (t, J=6Hz, 1H); 6.54 (d, J=7,6Hz, 2H); 6.72 (t, J=7,6Hz, 1H); 6.81 (d, J=7,8Hz, 1H); 7.04 (m, 3H); 7.18 (d, J=6,3Hz, 1H); 7.41-7.52 (m, 4H); 7.80 (d, J=7,9Hz, 1H); 7.90 (m, 1H); 8.11 (m, 1H); 9.50 (s, 1H).

31) 4.20 (s, br, 1H); 4.74 (d, J=2,8Hz, 2H); 6.68 (m, 3H); 7.16-7.27 (m, 4H); 7.38 (t, J=7Hz, 1H); 7.61 (d, J=9Hz, 1H); 7.69 (d, J=8Hz, 1H); 7.85 (d, J=8,4Hz, 1H); 9.52 (s, br, 1H).

Aa1) 2.23 (s, 3H); 3.63 (s, 2H); 3.99 (s, 2H); 7.2-7.6 (m, 6H); 7.80-7.90 (m, 2H); 8.16 (m, 2H); 8.30 (m, 1H).

Ab1) 2.16 (s, 3H); 3.50 (s, 2H); 3.60 (br, 2H); 3.88 (s, 2H); 6.65 (m, 2H); 7.16 (m, 2H); 7.38-7.48 (m, 4H); 7.74-7.84 (m, 2H); 8.20 (m, 1H).

Ba1) 3.99 (s, 2H); 4.25 (s, 2H); 7.39-7.57 (m, 6H); 7.74-7.89 (m, 2H); 8.08-8.20 (m, 3H).

1) CDCI3 2) sMSO-d6

Claims

Ciaims: 1. Compounds of the formula

wherein R1, R2 and Rg are the same or different and represent hydrogen, hydroxy, trifluormethyl, alkyl, alkoxy, acyloxy, halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, and R4 represents hydrogen or a group of formula

wherein R5 represents hydrogen or alkyl and R6, P7 and Ra are the same or different and represent hydrogen, hydroxy, trifluormethyl, alkyl, alkoxy, acyloxy. halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, in free form or, where such forms exist, in acid addition salt form.
2. Process for the production of compounds of formula

wherein R1, R2 and R3 are the same or different and represent hydrogen, hydroxy, trifluormethyl. alkyl, alkoxy, acyioxy, halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, and R4 represents hydrogen or a group of formula

wherein R5 represents hydrogen or alkyl and R6, R7 and R8 are the same or different and represent hydrogen, hydroxy, trifluormethyl, alkyl, alkoxy, acyloxy, halogen or benzyloxy, or two of these substituents together with the carbon atoms to which they are attached form an aliphatic, heteroaliphatic, aromatic or heteroaromatic ring, in free form or, where such forms exist, in acid addition salt form, which comprises hydrogenating a compound of formula

wherein the substituents are as defined above, whereby in this reaction functional groups may be protected by suitable protecting groups. which may be removed subsequent to the reaction in conventional manner, and recovering the compounds thus obtained in free form or in acid addition salt form.
3. A pharmaceutical composition comprising a compound according to claim 1 in free form or as appropriate in pharmaceutically acceptable salt form together with a ieast one pharmaceutically acceptable carrier or diluent.
4. A process for the preparation of a pharmaceutical composition according to claim 3 comprising mixing the compound with at ieast one pharmaceutically acceptable carrier or diluent.
5. A compound according to claim 1 in free form or as appropriate in pharmaceutically acceptable salt form for use as a pharmaceutical.
6. A compound according to claim 1 in free form or as appropriate in pharmaceuticaily acceptable salt form for use as PKC-inhibitors in the treatment of inflammatory diseases such as arthritis, in the treatment and prophylaxis of immunological disorders, of allergies, of skin diseases, in transplantations and in the field of oncology.
7. A method of treatment of a compound according to claim 1 in free form or as appropriate in pharmaceutically acceptable salt form for use as PKC-inhibitors in the treatment of inflammatory diseases such as arthritis, in the treatment and prophylaxis of immunological disorders, of allergies, of skin diseases, in transplantations and in the field of oncology, comprising admini stering a therapeutically effective amount of a compound according to claim 1 in free form or as appropriate in pharmaceutically acceptable salt form to a subject in need of such treatment.