DK151023B - METHOD OF ANALOGY FOR THE PREPARATION OF APOVINCAMINOL-3 ', 4', 5'-TRI-METHOXYBENZOATE DERIVATIVES - Google Patents

Description

in 151023

The present invention relates to an analogous process for the preparation of apovincaminol-3 ', 4', 5'-trimethoxybenzoate derivatives of the general formula I

Wherein R1 and R2 are both hydrogen or one of the symbols R1 and R2 represents hydrogen and the other represents nitro or pharmaceutically tolerable acid addition salts thereof, which process is characterized by the reaction of apovincaminol or an acid addition salt thereof. with 3,4,5-trimethoxybenzoic acid or a reactive acylating derivative thereof, whereby compounds of the formula la ΟςψψH, Ia CH3 °W «OCH₂ are isolated, or converted into a pharmaceutically tolerable acid addition salt thereof, or, if desired, nitrated, whereupon the mixture obtained by nitration mixes the compounds of formula Ib and le Νθ2 2 151023

"" VYS

oy) 2η5 ςΐΐ ° ννί lc cwV 0 CH30 is separated into the two constituents and a prepared compound of formula Ib or le, if desired, is converted into a pharmaceutically tolerable acid addition salt thereof.

The acid addition salts of the compounds of formula I can be formed with inorganic or organic acids. From the salts formed with inorganic acids, the hydrochlorides, sulfates and phosphates are particularly useful, as are the hydrogen tartrates, succinates, citrates and ascorbates from the salts formed with organic acids.

3 151023

It is known that the apovincaminol and its acetate have an effect on the corona artery (cf. French Patent No. 2,035,784). It is also known that the apovincaminol benzoate possesses general vasodilator properties (cf. Hungarian Patent No. 166,476; CA 82, 129 5,279 v (1975)) and that the esters of apovincaminol formed with alkanecarboxylic acid having 3-12 carbon atoms, have a cerebral vasodilatory effect (cf. Hungarian Patent No. 171,662, corresponding U.S. Patent No. 4,108,996 and German Patent No. 26,322,118).

Thus, all the known esters of apovincaminol have vasotropic action. However, the novel compounds of the present invention inhibit the enzyme activity of phosphodiesterase and may be used primarily to treat skin diseases associated with pathological cell proliferation and to prevent new onset of such diseases.

15 Diseases associated with the pathological growth of the epidermis are relatively frequent and comprise a few percent of the population.

These diseases can be both benign and malignant such as psoriasis atopias dermatitis, primary contact dermatitis, allergic contact dermatitis, baso- and spinocellular carcinoma, ichthyosis, premalignant hyperkeratosis, light-induced keratosis, acne and seborrhoea dermatitis. Some diseases occur only in humans, while others occur in both humans and animals.

Since some of the skin diseases associated with pathological cell growth (e.g., psoriasis) do not occur in animals, the effect of the compounds against psoriasis can only be indirectly demonstrated in animal experiments.

Voorhees et al. (Arch. Derm. 704, 359-365, (1971)) have demonstrated that a pathological cell proliferation is accompanied by a decrease in the level of cyclic adenozine monophosphate (c-AMP). It is known that c-AMP 30 is formed under the action of adenyl cyclase and is degraded by phosphodiesterase. Voorhees has succeeded in affecting psoriasis by agents that stimulate the function of adenyl cyclase (such as norepinephrine) or inhibit the function of phosphodiesterase (e.g. papaverine).

4 151023 J planning a model experiment test is based on the assumption that Voorhees's statements are also relevant in the opposite direction.

Thus, if it can be proven that a particular compound inhibits the function of phosphodiesterase, it is indirectly likely that this compound is suitable for the treatment of skin diseases associated with pathological cell proliferation.

This assumption has proven to be true; compounds that have phosphodiesterase inhibitory effect in in vitro tests have also been shown to be effective in the treatment of psoriasis in clinical experiments.

The model tests are performed using phosphodiesterase, which is isolated from animal body tissue [rat brain, bovine brain and bovine heart]. The enzyme is isolated by that of J. Schroder and H.V. Richenberg (Biochem. Biophys. Acta 302, 50 (1973)) described the isolated phosphodiesterase purified by the J.G. Hardman and E.W. Sutherland (J. Biol. Chem. 240, 3704 (1965)), and finally the activity of the purified enzyme is measured by the radioisotope method described by G. Poch in the presence of an excess of tritiated c-AMP (10.1 millimoles of c. AMP substrate, of which 3H-C-AMP is 2.59 K Bq) in an incubation system first without an inhibitor and then - in the presence of apoquinaminol-3r, 4 ', 5'-trimethoxybenzoate derivative as an inhibitor after 20-minute incubation period (NS Arch. Pharmacol. 268, 272 (1979)). From the test compound, a 1 millimolar stock solution is prepared and varying amounts are added to the incubated enzyme preparation 25 using this stock solution so that the concentration of the test compound in the incubated sample corresponds to 5 x 10, 1 x 10, 5 x 10, 1, respectively. x 10, 5 x 10 and 10 mol / liter. The aqueous solution of the compound used for comparison (papaverine) is added to the enzyme which is prepared in a similar manner.

The effect of the control sample (enzyme solution containing no inhibitor) is given the value 100%, while the effect of the solutions containing the compounds of the general formula I as papaverine is expressed as a percentage of the control sample. The results measured on the enzyme isolated from rat brains are shown in the table below.

Table 5

Test compound Concentration of test compound, (enzyme inhibitor) mol / liter 5 - 5 x 10 1 x 10 5 x 10

Effect on enzyme activity as a percentage of the control sample 10 Apovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 64.5 49.2 47.2 (-) - 11-Nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 53.6 44.4 37.3 (-) - 9-Nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 62.9 61.6 37.7

Papaverine 91.2 89.7 60.5

The results for the enzyme isolated from bovine brain and bovine heart are similarly measured. Using the results obtained, the enzyme activity is calculated based on the logarithm of the enzyme inhibitor concentration (expressed in ymol). The concentration of the enzyme inhibitor which reduces the enzyme activity by 50% (1Q value) is read by the curve. The results obtained are shown in the table below.

Table 6 151023 values in ymoles of phospho-Test compound diesterase enzyme isolated from 5 bovine heart rat brain

Apovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 1.5 (-) - 11-Nitro-apovincamino-3 ', 4', 5'-trimethoxybenzoate. HCl 18 - (-) - 9-Nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 1 15

Papaverin. HCl 50 70 15 The above table shows that the novel compounds of the invention are 33-50 and 4.5-9 times more effective, respectively, on the enzyme isolated from bovine heart and rat brain than the papaverine used as a reference compound.

The first clinical tests were performed with topical preparations containing the active ingredient (ointment, cream, solution, tincture, paste and aerosol). Creams containing 2%, 1%, 0.5%, 0.25% and 0.1% of (-) - 9-nitro- or (-) - 11-nitro-apo-vi respectively were used. caminol -3 ', 4', 5- 'trimethoxybenzoate.

Patients suffering from psoriasis are treated. A basic selection criterion is that patients do not simultaneously receive systemic treatment of their basic disease (e.g., immunosuppressive, cytostatic or glucocorticoid therapy).

Groups, each consisting of five patients, are investigated by the so-called piaque method. One side of the symmetrical skin lesions is treated with the cream containing the active ingredient, while the other side is treated with a placebo. The other affected skin surfaces of the patient are treated by other topical methods, including an ointment usually used for the treatment of psoriasis and containing flumethasone pivalate and salicylic acid, which ointment is used as a reference preparation.

The test begins with creams that have a high active ingredient content, and additional patients are treated with a cream with the lowest active ingredient content but still effective. Treatment continues for two weeks, twice a day, in an open dressing.

The effect is assessed by observing three different symptoms, namely inflammation, infiltration and desquamation (peeling). The strength of the symptoms is expressed on the scale below between 0 and 30 0 = no symptoms 15 1 = moderate symptom 2 = severe symptom 3 = very severe symptom.

Symptoms are assessed before treatment (I), after treatment for seven days (II) and after treatment for fourteen days (III). The following table shows the average number of points (the total number of points divided by the number of patients). A cream containing 2% active ingredient is used.

Table 8 151023

Average number of points

Test compound Infiltration Inflammation Desquamation

I II III I II III I li III

5 ----

Compound of formula Ia 1.8 0.6 0.5 1.8 1.3 0.75 1.2 0.4 0

Compound of Formula Ib 2.2 1 0.7 2.5 2.5 1.3 1.5 0.5 0 10 Compound of Formula Le 2.4 1.8 1.3 2.2 1.3 1.3 1.6 1.2 0.8

The process of the invention is preferably carried out in the presence of an organic solvent, in particular a chlorinated hydrocarbon or an aliphatic ketone or pyridine, in particular methylene chloride, chloroform or acetone. If an 3,4,5-trimethoxy-benzoyl halide is used as the acylating agent, the reaction is carried out in the presence of an equimolar amount or a small excess of an acid binder. For this purpose, for example, an alkali metal carbonate, alkali metal hydrogen carbonate 20 or an organic amine may be used. If 3,4,5-trimethoxybenzoic acid is used as the acylating agent, the reaction is carried out in the presence of a catalytic amount of an acid, preferably hydrochloric or sulfuric acid, or an activator of the carboxy group and / or a dehydrating agent. The carboxy group may be activated by a halogenated phenol, preferably pentachiorphenol. As dehydrating agent, for example, N, N'-di-cyclohexylcarbodimide may be used. The acylation may be carried out at a temperature between -20 ° C and the boiling point of the reaction mixture, preferably 20-60 ° C.

The compound of formula Ia can be isolated from the reaction mixture by extraction and / or evaporation.

9 151023

The product thus obtained can be converted into a pharmaceutically tolerable acid addition salt as indicated. Salt formation can be carried out by using an inorganic or organic acid (e.g. hydrochloric, sulfuric or phosphoric or tartaric, succinic, citric or ascorbic).

5 Salt formation is carried out in a manner known per se. It is preferable to proceed by adding a solution of the acid in ethyl ether or acetone to the solution of the base. Salt formation is achieved at a pH of 3-6.

The compound thus obtained of formula Ia may be nitrated as indicated if desired. Nitration can preferably be carried out by means of concentrated nitric acid. The reaction is preferably carried out in glacial acetic acid as a medium. The nitration is preferably carried out under cooling at a temperature of approx. 0 ° C. When the reaction is complete, the excess acid is neutralized and the thus obtained mixture of the compounds of formula Ib and le is isolated from the reaction mixture by extraction and / or evaporation.

The isomer mixture thus obtained is separated into the two components. The separation of the isomers can preferably be carried out by chromatography.

The compounds of formula Ib and le may, if desired, be converted into pharmaceutically tolerable acid addition salts thereof by the methods described in connection 20 with the preparation of the acid addition salts of the compound of formula la.

The method of the invention allows for the provision of pharmaceutical compositions with phosphodiesterase inhibitory effect which can be used mainly for the treatment of skin diseases associated with pathological cell proliferation, and the prevention of new attacks of these diseases, these compositions comprising the compound of the active ingredient. general formula I or a pharmaceutically tolerable acid addition salt thereof and optionally additional therapeutically effective compounds in a mixture with conventional pharmaceutical carriers and / or diluents.

The active ingredient content of the pharmaceutical compositions of the invention is preferably 0.1-8.0%, especially 0.2-2.0%. The compositions may optionally further contain pharmaceutically and therapeutically active compounds such as antibiotics, cytostatic agents, prostaglandins, ditranol, salicylic acid, tar, anti-inflammatory agents, immunosuppressants, glucocorticoids and - in the case of preparations suitable for parenteral administration - topical drugs. As a glucocorticoid, triamcinolone-acetonide can preferably be used. The active ingredient may preferably be pre-treated in the form of topical preparations such as creams, ointments, solutions, jellies, aerosols, aerosol foams or adhesive patches.

The active ingredient may preferably be used in the form of the base, but acid addition salts may also be used.

. It is preferred to incorporate the active ingredient into a cream which can be washed.

Creams can be prepared by dissolving the active ingredient in an alcohol-type solvent, preferably in propylene glycol or ethylene glycol or a mixture thereof formed with a small amount of water, and mixing the thus obtained solution with a fat phase which can be easily applied and is skin compatible.

The fat phase may include cetyl alcohol, stearyl alcohol, cetostearyl alcohol, paraffin oil or glycerine monostearate.

The cream may also contain an emulsifier, preferably polyoxyethylene sorbitan monoosteate or monostearate, and a preservative such as benzoic acid derivatives, preferably methyl p-hydroxybenzoate.

The creams may preferably contain 0.25-2.0% active ingredient, 45-50% glycol, 23-27% paraffin oil, 11-15% stearyl alcohol and optionally other adjuvants up to 100%.

The active ingredient may also be formulated in the form of an ointment which cannot be washed off with water by incorporating the active ingredient directly into the fat phase.

11 151023

The active ingredient may also be formulated in the form of a solution or tincture which may contain, for example, 20-40% propylene glycol or dipropylene glycol, 40-55% 96% ethanol and distilled water in up to 100%.

The aerosol formulations can be prepared by adding to the solution of the active ingredient in propylene glycol a fat, e.g., isopropyl myristate, and a propellant, e.g., Freon.

Injectable solutions suitable for parenteral administration and preferably useful by subcutaneous or intracutaneous route can be prepared by dissolving a salt of the active ingredient in a 0.72% 10 aqueous sodium chloride solution and adjusting the pH of the solution to 5th

The pharmaceutical compositions can be prepared by methods known in the pharmaceutical industry. It can be proceeded by mixing the active ingredient and optionally additional therapeutically active compounds with suitable inert non-toxic known pharmaceutical carriers and / or additives and finalizing a thus obtained mixture until it has a form suitable for medical use.

The invention is further illustrated by the following examples.

EXAMPLE 1

Preparation of (-j-apovincaminol-5 '' - trimethoxybenzoate 3.10 g (10.1 millimoles) (-) - apovincaminol is dissolved in 60 ml of anhydrous dichloromethane, then 3.10 g of anhydrous sodium carbonate and 2, 50 g (10.9 millimoles) of 3,4,5-trimethoxybenzoyl chloride and the reaction mixture is stirred at room temperature for 24 hours. The reaction mixture is diluted with 100 ml of water, the organic phase is separated and the aqueous phase is extracted twice with each time. The combined dichloromethane phases are dried over magnesium sulfate, filtered and the filtrate is evaporated in vacuo to give 4.50 g of the title compound, yield 89.1%.

12 151023

Gross formula ^ 39 ^ 34 ^ 2 ^ 5- Molecular weight 502.61.

IR Spectrum (film): v = 1725 (-C = O) and 1620 (= C = C =) cm

max NMR Spectrum (CDCl3): δ (ppm) = 1.01 (t, 3H, CA / g 3.72 (s, 6H, 2 x -OCH 3); 3.85 (s, 3H, -OCHg); 4.25 (s, 1H, annealing); 5.29 (s, 1H, -CH =); 5.4 (m, 2H, -OCl · +); 7.0-7.8 (m, 6H, aromatic).

Mass spectrum: m / e = 502 (53), 432 (100), 290 (17), 261 (41), 220 (19), 216 (23), 212 (18), 195 (35).

[α] D = -22.0 ° (c = 0.7; CH 2 Cl 2).

EXAMPLE 2

Preparation of (-) - apovincaminol-3 ', 4', 5'-trimethoxybenzoate hydrogen tartrate

A compound prepared in Example 1 is dissolved in diethyl ether. To the solution is added a saturated solution of D-tartaric acid in diethyl ether until the precipitation of the hydrogen tartrate salt is complete. The salt is filtered off and dried. Melting point, 120-121 ° C.

IR Spectrum (KBr): v = 1730 (-C = 0) and 1640-1665 (= C = C =) cm

ΓΠ3Χ [a] + = -8.5 ° (c = 1 in pyridine)

Molecular Weight 652.7.

EXAMPLE 3

Preparation of (-10-nitro-apovincaminol-5'-5'-trimethoxybenzoate and (-) - 11-nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate g (-) - apovincaminol-3 ', Dissolve 4 ', 5'-trimethoxybenzoate in 50 ml of glacial acetic acid. To the solution obtained, add a mixture of 20 ml glacial acetic acid 25 and 10 ml of concentrated nitric acid (d - 1.52) at 0 ° C with stirring. water and the pH is adjusted to 9 by adding a 25% solution of aqueous ammonium hydroxide. The alkaline solution is first extracted with 250 ml and then twice with 200 ml of dichloromethane each time. The total organic solution is dried. over anhydrous sodium sulfate and filtered and the filtrate evaporated to dryness in vacuo.

An isomer mixture thus obtained is separated into the two components by chromatography using silica gel 60 as adsorbent and a 10: 2 mixture of benzene and acetone as the developing agent. Under these conditions, the R ^ value of the 9-nitro compound is greater than the R ^ value of the 11-nitroderate. Thus, 1.7 g of the 9-nitro compound and 1.8 g of the 10-11 nitro derivative are obtained.

The physical constants of the (-J-O-Nitro-apovincaminol-S '^' / S 'trimethoxybenzoate are as follows, m.p. 77-78 ° C.

[α] D = -78.5 ° (c = 1 in CHCl 3).

1 H-NMR Spectrum (CDCl 3): δ (ppm) = 3.8 (s, 6H, 2 x CH 3 O); 3.91 (s, 3H, 1 x CH 3 O-); 7.3 (d, 1H, H12); 7.96 (d, 1H, Hn); 8.1 (d, 1H, H10).

The physical constants of the (-) - 11-Nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate are as follows, m.p. 72-73 ° C.

[α] D = -134.3 ° (c = 1 in CHCl 3).

1 H NMR Spectrum (CDCL 3): δ (ppm) = 3.8 (s, 6H, 2 x CH 3 <>); 3.9 (s, 3H, 1 x CH 3 O); 7.56 (d, 1H, Hg); 8.1 (d, 1H, H1 ()); 8.9 (d, 1H, H12).

EXAMPLE 4

Preparation of O-Nitro-Apovincaminol-S '^^ S'-trimethoxybenzoate hydrochloride and 11-nitro-apovincaminol-3', 4 ', 5'-trimethoxybenzoate hydrochloride 5 The 9-nitro prepared in Example 3 - or 11-nitro-apovincaminol-S '^ jSS trimethoxybenzoate is dissolved in methanol and the pH of the solution is adjusted by adding a solution of hydrogen chloride and methanol. The hydrochloride salt formed is precipitated by the addition of diethyl ether. The salt is filtered off, washed and dried. The 9-nitro derivative hydrochloride has a melting point of 144-150 ° C, while the 11-nitro derivative hydrochloride has a melting point of 141-144 ° C.

EXAMPLE 5

A cream of the following composition is prepared

Ingredient Quantity (g) 15 - -

Apovincaminol-3 ', 4', 5'-trimethoxybenzoate 2

Propylene Glycol 50

Paraffin Oil 26

Polyethylene glycol 5 20 S tea ry lal carbon 15

Glycerol Monostearate 2

The active ingredient is dissolved in propylene glycol in a water bath (since the temperature of the bath does not exceed 50 ° C). The other ingredients are heated until melted and then cooled to 40-45 ° C with constant stirring. To the melted ingredients, the solution of the active ingredient is added, with stirring, and the cream thus obtained is cooled with stirring.

Similarly, creams containing 0.25%, 0.5%, 1.0% and 1.5% of the active ingredient are prepared, respectively.

in 151023

The active ingredient 9-nitro- or H-nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate can also be used.

EXAMPLE 6

A cream of the following composition is prepared. Component Amount (g) 9-Nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate 2

Triamcinolone Acetonide 0.1 Glycerol Monostearate 3.0

Polyethylene Glycol 400 5.0

Stearyl alcohol 13.0

Paraffin Oil 24.9

Propylene Glycol 53.0 15 Proceed in the same manner as in Example 5 except that two active ingredients are dissolved in propylene glycol. The 11-nitrode derivative or apovincaminol-3 ', 4', 5'-trimethoxybenzoate can also be used as active ingredient.

Example 7 20 Preparation of a tincture solution of the following composition

Ingredient Amount (g) 9-Nitro-apovincaminol-3,4,4,5'-trimethoxybenzoate 1 25 Propylene glycol 30 96% methanol 69 16 151023

The above process can also be carried out using the 11-nitro derivatives as the active ingredients.

EXAMPLE 8

A tincture solution of the following composition is prepared 5 Component Amount (%)

Apovincaminol-S '^^ S'-trimethoxybenzoate hydrogen tartrate 1

Propylene Glycol 30 10 96% Ethanol 47

Distilled water 22 EXAMPLE 9

An aerosol of the following composition is prepared

Ingredient Quantity (%) 15 - -

Apovincaminol-3 ', 4, 5, -trimethoxybenzoate hydrogen tartrate 0.5

Propylene glycol 30 in sopropyl myristate 4.5 20 Freon 65 9-Nitro or 11-nitro derivative may also be used as active ingredient.

Claims (8)

EXAMPLE 10 An aerosol foam of the following composition is prepared Component Amount (%) An analogous process for the preparation of apovincaminol-S '^^ S'-tri-methoxybenzoate derivatives of the general formula I R1 ΛΑΑΚ OCH3 wherein R1 and R2 are both hydrogen or one of the symbols R1 and R2 represents hydrogen and the other represents nitro, or pharmaceutically tolerable acid addition salts thereof, characterized in that apovincarmnol or an acid addition salt thereof is reacted with 3,4,5-trimethoxybenzoic acid or a reactive acylating derivative thereof, whereby compounds of the formula la Øgffi, S * Xj ° OCH3 are isolated, or is converted into a pharmaceutically tolerable acid addition salt thereof or, if desired, nitrated, and the mixture obtained by nitration of the compounds of formula Ib and le NO2 CH3O 151022 jOcXii ^^ 5 CH3 ° v ^ Y ^ y le α # τγ Cttf) is separated into the two constituents and, if desired, a compound of formula Ib or le prepared into a pharmaceutically tolerable acid addition salt thereof. Process according to claim 1, characterized in that apovincaminol-3, 4 ', 5'-trimethoxybenzoate or pharmaceutically tolerable acid addition salts thereof are prepared. Process according to claim 1, characterized in that 9-nitro-apovincaminol-10 3 ', 4', 5'-trimethoxybenzoate or pharmaceutically tolerable acid addition salts thereof are prepared. Process ... according to claim 1, characterized in that 11-nitro-apovincamino-3 ', 4', 5'-trimethoxybenzoate or pharmaceutically tolerable acid addition salts thereof are prepared. Process according to claim 1, characterized in that 3,4,5-trimethoxy-benzoyl chloride is used as an acylating agent. 151023 5 Apovincami nol -3 ', 4', 5 'trimethoxy benzoate hydrogen tartrate 2 Cetostearyl alcohol 1 Benzyl alcohol 2 Polyoxyethylene sorbitan monostearate 15 10 96% ethanol 30 Distilled water 30 Freon 20 Process according to claim 1 or 5, characterized in that the acylation is carried out in the presence of an organic solvent. Process according to claims 1, 5, characterized in that the nitration is carried out in the presence of glacial acetic acid. Process according to claim 1 or 7, characterized in that the nitration is carried out at a temperature of approx. 0 ° C.