SE449863B - APOVINCAMINO DERIVATIVES, PREPARATION AND PHARMACEUTICAL PREPARATION THEREOF - Google Patents

Description

(44) 863 to 2 (Ib) CH 3 O CH 3 O CH 3 O and 11-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate of the formula OZN (IC) CH 3 O \\ 0: 0 CH 3 O CH 3 O as well as also acid addition salts thereof.

In the general formula I, either R 1 and R 2 represent hydrogen or one of the symbols R 1 and R 2 represents hydrogen and the other nitro.

The acid addition salts of the compounds I may be formed with inorganic or organic acids. Among salts with inorganic acids, the hydrochlorides, sulphates and phosphates are particularly useful, and among the salts with organic acids, hydrogen tartrate, succinate, citrate and ascorbate are particularly useful.

It is known that apovinaminol and its acetate have an effect on the coronary arteries (French Pat. No. 2,035,784). It is also known that apovinquinol benzoate has quite general vasodilatory properties (Hungarian Patent Specification 166,476; CA 82, 129,279 v / 1975 /) and the esters of apovinquinol formed with alkane carboxylic acids having 3 to 12 carbon atoms have a cerebral vasodilatory effect (Hungarian Patent 171,662, the corresponding U.S. Patent 4,108,996 and the BR West German Patent 26 32,118).

Thus, all known esters of apovinkaminol show vasotropic effects. On the other hand, the novel compounds of the present invention inhibit the enzyme activity of phosphodiesterase and can be used primarily for the treatment of skin diseases which accompany pathological cell proliferation and as prophylaxis against recurrence of such diseases.

Diseases that accompany pathological proliferation of the epidermis are relatively common and affect a few percent of the population. These diseases can be of both benign and malignant nature, such as psoriatic atopidermatitis, primary contact dermatitis, allergic contact dermatitis, baso- and spinocellular carcinoma, inchthyosis, premalignant hyperceratosis, easily induced seratosis, acne and seborrheic dermatitis. Some diseases only affect humans, while others affect both humans and animals.

Since some of the skin diseases that accompany pathological cell proliferation (eg psoriasis) do not occur in animals, the activity of compounds against psoriasis can only be indirectly detected by animal experiments.

Voorhees o.a. (Arch. Derm. 104, 359-365 / 1971 /) have found that pathological cell proliferation is accompanied by a decrease in the level of cyclic adenosine monophosphate (c-AMP). It is known that c-AMP is formed under the action of adenyl cyclase and degraded by phosphodiesterase. Voorhees succeeded in affecting psoriasis with agents that stimulate adenyl-cyclase function (such as nor-epinephrine) or inhibit phosphodiesterase function (eg papaverine).

When planning the model experiment test, it has been assumed that Voorhees' statement is relevant as well as its inversion. Thus, if it can be proved that a certain compound inhibits the function of the phosphodiesterase race, this indirectly makes it probable that said compound is suitable for the treatment of skin diseases which accompany pathological cell proliferation.

This proved to be true: compounds that exhibited phosphodiesterase inhibitory activity in in vitro tests were also found to be active in the treatment of psoriasis in clinical trials.

The model tests performed using phosphodiesterase isolated from animal tissues (brain from rats and brain and heart from cattle). The enzyme was isolated according to the method described by J. Schröder and H.V. Richenberg (Biochem. Biophys. Acta 302, 50/1973 /) and the isolated phosphodiesterase were purified according to the method developed by J.G. Hardman and E.W.

Sutherland (J. Biol. Chem. 240, 3704/1965 /) and finally the activity of the purified enzyme was measured according to the radioisotope method prepared by G. Pöch in the presence of an excess of tritiated c ~ AMP (10, 1 mmol c-AMP substrate, of which 3H-c-AMP is 2.59 K Bq) in an incubation system first without inhibitor and then in the presence of apovincaminol 3 ', 4', 5'-trimethoxy-benzoate derivatives as an inhibitor after an incubation period of 20 minutes (NS Arch. Pharmacol. 268, 272/1979 /). From the test compound a 1 millimolar stock solution was prepared and to the incubated enzyme preparations different amounts were added by means of said stock solution, so that the test compound in the incubated sample corresponding to 5 x 10-7; 1 x 10-6; 5 x 10%; 1 x 104; 5 x 1o "5 moi / liter.

The aqueous solution of the compound used for the comparison and 10_k (papaverine) was added to the enzyme prepared in the same manner.

The activity of the control sample (enzyme solution which did not contain any inhibitor) was designated as 100%, whereas the activity of the solutions containing the compounds I as papaverine is indicated as a percentage of the control sample. The resulting measurements of enzyme isolated from rat brain are summarized in the following table. 10 15 20 25, 30 35 40 il 449 863 Test compound Test compound concentration (enzyme inhibitor) mol / liter s x10 '° 1x1o'5 s x1o "s (Effect on enzyme activity, percentage of control) apovinkaminol-3', 4 ', 5'-trimethoxy-benzoate.HCl 64.5 49.2 47.2 (-) ~ 11-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate.HCl 53.6 44, 4 37.3 (-) - 9-nitro-apovincaminol-7 ', 4', 5'-trimethoxy-benzoate.HCl 62.9 61.6 37.7 Papaverine 91.2 89.7 60, The result on enzyme isolated from bovine brain and heart was measured in the same way, using the results obtained, the enzyme activity was plotted as a function of the logarithm of the enzyme inhibitor concentration (expressed in μmol) The concentration of the enzyme inhibitor which reduces the enzyme activity by 50% (Iso) was read on the curve and the results obtained are summarized in the following table.

Test compound Iso values in μmol of phosphodiesterase enzyme isolated from bovine heart rat brain apovincaminol-3 ', 4', 5'-trimethoxy-benzoate.HCl 1.5 10 (-) - 11-nitro-apovincaminol ~ 3 ', 4 ', 5'-trimethoxy-benzoate.HCl 1 8 (-) - 9-nitro-apovincaminol-3', 4 ', 5'-trimethoxy-benzoate.HCl 1 15 Papaverine.HCl 50 70 It appears from the results compiled in the above table that on the enzyme isolated from bovine creature 6 15 heart is seen and from rat brain the new compounds according to the present invention are 33-50 and 4.5-9 times respectively strong as the papaverine used as a reference substance. The first clinical trials were conducted with topical preparations containing the active ingredient (ointment, cream, solution, tincture, paste, aerosol). Creams containing 2%, 1%, 0.5%, 0.25% and 0.1% (-) - 9-nitro- and (-) - 11-nitro-apovincaminol-3 ', 4', 5'- trimethoxy benzoate was used. 7 I Patients with psoriasis were treated. An additional fundamental point of view in the choice was that the patient did not at the same time receive any systemic treatment of the underlying disease (for example immunosuppressive or cytostatic treatment or glucocorticoid treatment.

In groups of five patients each, each was examined by means of the so-called "plaque methods". One side of the symmetrical skin lesions was treated with the cream containing the active ingredient, whereas the other side was treated with placebo. The other affected skin surfaces of the patient were treated with other topical methods, e.g. with an ointment commonly used to treat psoriasis and containing flumethasone pivalate and salicylic acid. This ointment was used as a reference substance.

The test had been initiated with ointments with a high content of active ingredient and additional patients were treated with ointments that were still active but which contained lower levels of active ingredient. The treatment was carried out for two weeks twice daily in open dressings.

The effect was determined by observing three different symptoms, namely inflammation, infiltration and desquamation (peeling of the skin). The intensity of the symptoms was divided according to the following scale between 0 and 3. 0 = no symptoms 2 = severe symptoms f ”1 = moderate symptoms 3 = very severe symptoms The symptoms were estimated before the treatment (I), after the treatment for seven days (II) and after a treatment for 14 days (III). The table below indicates average points (the sum of points divided by the number of patients 10 15 20 25 30 35 40 449 863 patients). An ointment containing 2% active ingredient was used.

Test compound Average number of points Infiltration Inflammation Desquamation I II III I II III I II III Compound of formula Ia 1.8 0.6 0.5 1.8 1.8 0.75 1.2 0.4 Compound of formula Ib 2 , 2 1 0.7 2.5 2.5 1.3 1.5 0.5 o Compound of formula Ic 2.4 1.8 1.3 2.2 1.3 1.3 1.6 1.2 0.8 According to the invention there is also provided a process for the preparation of the novel compounds I, wherein either both R 1 and R 2 represent hydrogen or one of the symbols R 1 and R 2 represents hydrogen and the other nitro, and pharmaceutically acceptable acid addition salts thereof, which process comprises reacting apovincaminol or an acid addition salt thereof with 3,4,5-trimethoxy-benzoic acid or a reactive derivative thereof which can be acylated, whereupon the compound of formula Ia is isolated or converted into a pharmaceutically acceptable acid addition salt thereof, or, if if desired, the compound of formula Ia is nitrated and the mixture of the compounds of formulas Ib and Ic thus obtained is separated into two components. enter and, if desired, a compound of the general formula Ib or Ic thus obtained is converted into a pharmaceutically acceptable acid addition salt. The process according to the invention is suitably carried out in the presence of an organic solvent, preferably a chlorinated hydrocarbon or an aliphatic ketone or. pyridine, especially in methylene chloride, chloroform or acetone. If a 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 slight excess of an acid-binding agent. For this purpose, for example, alkali metal carbonate, alkali metal bicarbonate or organic amine are used. If 3,4,5-trimethoxy-benzoic acid is used as acylating agent, the reaction is carried out in the presence of a catalytic amount of an acid (preferably hydrochloric acid or sulfuric acid) or an activator of the carboxyl group and / or a dehydrating agent. The carboxyl group can be activated with a halogenated phenol, preferably pentachlorophenol. As the dehydrating agent, for example, N, N'-dicyclohexylcarbodiimide was used. The acylation can be carried out at a temperature between -20 ° C and the boiling point of the reaction mixture, preferably between 20 and 60 ° C.

Compound Ia can be isolated from the reaction mixture by extraction and / or evaporation.

The product thus obtained can be converted into a pharmaceutically acceptable acid addition salt. The salt formation can be carried out using inorganic or organic acid (for example hydrochloric acid, sulfuric acid, phosphoric acid or tartaric acid, succinic acid, citric acid or ascorbic acid). The salt formation is carried out in a manner known per se. The acid dissolved in ethyl ether or acetone is preferably added to the solution of the base. Salt formation takes place at pH 3-6.

The compound Ia thus formed can, if desired, be nitrated. The nitration can preferably be carried out with the aid of concentrated nitric acid. The reaction is conveniently carried out in glacial acetic acid as a reaction medium. The nitration is suitably carried out under cooling at a temperature of approximately 0 ° C. After the reaction is complete, the excess acid is neutralized and the mixture of the compounds Ib and Ic thus formed is isolated from the reaction mixture by extraction and / or evaporation.

The isomer mixture thus obtained can be separated into two components. This is preferably accomplished by chromatographic methods.

Compounds Ib and Ic may, if desired, be converted into pharmaceutically acceptable acid addition salts in the manner set forth above for the preparation of the acid addition salts of compound Ia.

The invention also relates to pharmaceutical preparations having a phosphodiesterase inhibitory effect and the main utility of these preparations is for the treatment of skin diseases which accompany pathological cell proliferation and for the prevention of recurrence. These preparations contain as active ingredient a compound of the general formula I or a pharmaceutically acceptable acid addition salt thereof and optionally further therapeutically active compounds in admixture with Pharmaceutical carriers and / or diluents.

The active ingredients are included in the pharmaceutical preparations according to the invention, preferably 0.1-8.0, in particular 0.2-2.0%. The formulations may optionally contain other pharmaceutically therapeutically active compounds, such as antibiotics, cytostatics, prostaglandins, ditranol, salicylic acid, tar, anti-inflammatory agents, immunosuppressive agents, glucocorticoid and - where the formulations are suitable for parenteral administration - local anesthetics. As glucocorticoid, triaminolone acetonide is preferred. The active ingredient may be prepared, preferably, in the form of preparations for topical use, such as in the form of creams, ointments, solutions, gels, aerosols, aerosol foams, adhesive patches, etc.

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

It is preferred to mix the active ingredient in a cream which can be washed off.

The cream can be prepared by dissolving the active ingredient in an alcohol-type solvent, preferably in propylene glycol or ethylene glycol or in a mixture thereof with a small amount of water, then mixing the solution thus obtained with an easily applicable oily phase which is compatible with the skin.

Said fatty phase may be cetyl alcohol, stearyl alcohol, cetostearyl alcohol, paraffin oil, glycerol monostearate, etc.

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

The creams may preferably contain 0.25-2.0% of the active ingredient, 45-50% glycol, 23-27% paraffin oil, 11-15% stearyl alcohol and optionally up to 10 15 20 25 30 35 40 449 10 865 100% other aids.

This active ingredient can also be prepared in the form of an ointment which cannot be washed off with water. In this case, the active ingredient is mixed directly into the fatty phase. w) The active ingredient can also be prepared in the form of a solution or tincture, which may contain for example 20-40% propylene glycol or dipropylene glycol, 40-55% of a 96% ethanol and up to 100% distilled water .

The aerosol formulations can be prepared by adding to the solution of the active ingredient in propylene glycol a fatty substance, for example isopropyl myristate, and a propellant (for example freon).

Injectable solutions suitable for parenteral administration, preferably subcutaneously or intracutaneously, can be prepared by dissolving a salt of the active ingredient in a 0.72% aqueous solution of sodium chloride and adjusting the pH of the solution to 5.

The pharmaceutical preparations according to the invention can be prepared in ways which are applied in the pharmaceutical industry. It can be carried out by mixing the active ingredient and any further therapeutically active compounds with suitable, inert, non-toxic, known pharmaceutical carriers and / or additives and preparing the mixture thus obtained in a suitable form for medical use.

Further details regarding the present invention appear from the following examples, but the invention is of course not limited to the particular features and details set forth in the examples.

Example 1 Preparation of (-) - apovincaminol-3 ', 4', 5'-4,9-trimethoxy-benzoate 3.10 g (10.1 mmol) 60 ml of anhydrous dichloromethane, then 3.10 g of anhydrous (-) -apovincaminol was dissolved in sodium carbonate and 2.50 g (10.9 mmol) of 3,4,5-trimethoxy-benzoyl chloride were added and the reaction mixture was stirred at room temperature for 24 hours. The reaction mixture was diluted with 100 ml of water, the organic phase was separated and the aqueous was extracted with dichloromethane (2 x 20 ml). The combined dichloromethane phases were dried over magnesium sulphate, filtered and the filtrate evaporated in vacuo. 4.50 g of the title compound were thus obtained. Yield 89.1%.

Gross formula C30H36N2O5. Molecular Weight 502.61.

IR spectrum (film): νma 1725 cm -1 (-C = O); 1620 cm_1 (= C = C =). NMR Spectrum (deuterochloroform): δ: 1.01 (t, 3H, CH 3 CH 2 -); Δ: 3.72 (s, 6H, 2 x -OCH 2); Δ: 3.85 (s, 3H, -OCH 3); 6: 4.25 (s, 1H, annealing); Δ: 5.29 (s, 1H, -CH =); 6: 5.4 (m, ZH, -OCH 2); Δ: 7.0-7.8 (m, 6H, aromatic).

MS (m / e): 502 (53), 432 (100), 290 (17), 261 (41), 220 (19), 216 (23), 212 (18), 195 (35). tungs = -22.0 ° (C = 0.7; dichloromethane).

Example 2 X Preparation of (-) - apovincaminol-3 ', 4', 5'-trimethoxy-benzoate-hydrogen tartrate The compound prepared as in Example 1 was dissolved in diethyl ether. To the solution was added a saturated solution of D-tartaric acid in diethyl ether, until the hydrogen tartrate was completely precipitated. The salt was filtered off and dried. Melting point 120-121 ° C.

IR spectrum (KBr) = νmax 1730 cm -1 (-c = o); 1640-1665 cm -1 (= C = C =).

LQJÉS = -s, 5 ° (6 = 1; pyriain). Moiekyivikt 652.7.

Example 3 Preparation of (-) - 9-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate and (-) -11-nitro-apovincamino-3 ', 4', 5'-trimethoxy benzoate 5 g of (-) - apovincaminol-3 ', 4', 5'-trimethoxy benzoate were dissolved in 50 ml of glacial acetic acid. To the solution thus obtained was added a mixture of 20 ml of glacial acetic acid and 10 ml of concentrated nitric acid (d = 1.52) at 0 ° C with stirring. The reaction mixture was poured into 350 ml of ice-cold water and the pH was adjusted to 9 by adding a 25% aqueous solution of ammonium hydroxide. The alkaline solution was extracted with dichloromethane first 1 x 250 ml and then 2 x 200 ml. The combined organic solutions were dried over anhydrous sodium sulfate, filtered and the filtrate was evaporated to dryness in vacuo.

The isomer mixture thus obtained was separated into two components by chromatographic methods using "Kieselgel" 60 as adsorbent and with a mixture of benzene and acetone 10: 1 as developing agent. Under these conditions, the Rf value of the 9-nitro compound was greater than that of the 11-nitro derivative. 1.7 g of the 9-nitro compound and 1.8 g of the 11-nitro derivative were thus maintained.

The physical constants of (-) - 9-nitro--apovincaminol-3 ', 4', 5'-trimethoxy-benzoate were as follows: mp 77-78 ° C. tenn = -7s, 5 ° (e = 1; klereferm). 1 H-NMR (cDc13) = δ: 3.8 (e, en, 2 X cH, o-); se 3.91 (e, 3H, 1 x CH 3 O-); Δ: 7.3 (d, 1H, H12); Δ: 7.96 (d, 1H, H11); 6: 8.1 (d, 1H, H10).

The physical constants of (-) - 11-nitro--apovincaminol-3 ', 4', 5'-trimethoxy-benzoate are as follows: melting point 72-73 ° c; : still = -134.3 ° (e = 1; clothing). 1 H-NMR (δ, δ; 3.8 (ε, δ, 2 x cH 3 O-); δ: 3.9 (ε, δ, 1 X CH 3 O-); δ: 8.1 (d, 1H, H10); ε 8.9 (d, 1H, H12).

Example 4 Preparation of 9-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate hydrochloride and 11-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate-hydrochloride 9-nitro or 11-nitro-apovincaminol 3 ', 4', 5'-trimethoxy-benzoate prepared in the manner set forth in Example 3 was dissolved in methanol and the pH of the solution was adjusted by adding a solution of HCl in methanol. The hydrochloride formed precipitated upon addition of diethyl ether. The salt was filtered off, washed and dried. The hydrochloride of the 9-nitro derivative and the hydrochloride of the 11-nitro derivative had melting points of 144 DEG-150 DEG C. and 141 DEG-144 DEG C., respectively.

Example gel 7 7 A cream having the following composition was prepared: 10 15 20 25 30 35 40 13 449 865 Component Amount, grams of apovincaminol-3 ', 4', 5'-trimethoxy-benzoate 2 propylene glycol 50 paraffin oil 26 polyethylene glycol 5 stearyl alcohol 15 glycerol monostearate 2 The active ingredient was dissolved in propylene glycol in a water bath (water bath temperature not exceeding 500). The other components were heated to melt, then cooled to 40-45 ° C with continuous stirring.

To the melt was added the solution of the active ingredient with stirring, and the cream thus obtained was cooled with continued stirring.

In an analogous manner, creams containing 0.25 were prepared; 0.5; 1.0 and 1.5% active ingredient, respectively.

The active ingredient 9-nitro- or 11-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-benzoate may also be used.

Example Gel 6 A cream having the following composition was prepared: Component, Amount, Gram, 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 I 24.9 propylene glycol 53.0 It was carried out in a manner analogous to that given in Example 5 with the modification that two active ingredients were dissolved in propylene glycol. As the active ingredient, the 11-nitro derivative or apovincaminol-3 ', 4', 5'-trimethoxy-benzoate can also be used.

In Example 7 A tincture having the following composition was prepared: Component 1 Amount, amount of 9-nitro-apovincaminol-3 ', 4', 5'-trimethoxy-1-benzoate propylene glycol 96 % methanol _ 69 The process described above can also be carried out using the 11-nitro derivatives as active ingredient.

Example gel 8 A tincture having the following composition was prepared: Component - Amount,% apovincaminol-3 ', 4', 5'-trimethoxy-benzoate-hydrogen tartrate 1 propylene glycol 96% ethanol 47 distilled water 22 Example gel 9 An aerosol having the following composition was produced:.

Component Amount,% apovincaminol-3 ', 4', 5 '- trimethoxy-benzoate-hydrogen tartrate 0.5 propylene glycol 30 isopropyl myristate 4,5 "Freon" 65 The .9-nitro- or 11-nitro derivative can also be used as active ingredient. .

Example Gel An aerosol foam having the following composition was prepared: Component Amount,% 'Apovincaminol-3', 4 ', 5' - trimethoxy-benzoate-hydrogen tartrate 2 cetostearyl alcohol _ 1 benzyl alcohol polyoxyethylene sorbitan monostearate. 95% ethanol - 30 distilled water 30 "Freon" _ 20 NP 'w

Claims (6)

449 863 15 PATENT CLAIMS Compounds of general formula 5 (I) 10 cnso lf O CH 3 O ÛCH 3 wherein either R 1 and R 2 both represent hydrogen or one of the symbols R 1 and R 2 represents hydrogen and the other nitro, and pharmaceutically acceptable acid addition salts thereof. 20 Apovincaminol-3 ', 4', 5'-trimethoxy-benzoate according to claim 1 of the formula CH2 / C25 (Ia) O /// CHaO Ü 30 CH3O 10 and pharmaceutically acceptable acid addition salts hence. Α-nitro-apovincaminol-3 ', 4', 5'-trimethoxybenzoate according to claim 1 of the formula (Ib) CH 3 O 10 CH 3 O CH 3 O and pharmaceutically acceptable acid addition salts thereof. An 11-nitro-apovinquinol-3 ', 4', 5 '-trimethoxy-benzoate according to claim 1 of the formula OZN (Ic) CH 3 O Q CH 3 O 3 CH 3 O and pharmaceutically acceptable acid addition salts thereof. M 10 15 20 25 30 35 449 863 17 A process for the preparation of novel compounds according to claim 1 having the general formula - R 2 C 2 H 5 (I) CH 3 O 2 CH 3 O CH 2 wherein either R 1 and R 2 both represent hydrogen or a 1 and R 2 represent hydrogen and the other of the symbols R acid addition salts thereof, characterized by reacting apovincaminol or an acid addition salt thereof with 3,4,5--trimethoxy-benzoic acid or a reactive derivative thereof which can effect acylation, preferably 3,4,5-trimethoxy-benzoyl chloride, suitably in the presence of an organic solvent, whereupon the compounds of formula (Ia) are isolated CH 3 O 2 CH 3 OCH 2 OCH 2 or converted into pharmaceutically acceptable acid addition salts, or, if desired, the compound is nitrated with the formula (Ia), preferably at a temperature of about 0 DEG C., then separating the mixture of the compounds so obtained with the formulas NO2 (Ib) CH2 O WO CH3O CH3O and O2N (IC) /, / c two components, and, if desired, converting a compound of the general formula (Ib) or (Ic) thus obtained into a pharmaceutically acceptable acid addition salt. Pharmaceutical preparation for the treatment of diseases which accompany pathological cell proliferation, in particular psoriasis, and for the prevention of recurrence of such diseases, characterized in that it is an active ingredient, preferably in an amount of 0.1-8.0, more preferably 0.2-2.0%, contains a compound according to claim 1 of the general formula (I) CH 3 O CH 3 O ÛCH 3 wherein R 1 and R 2 have the meanings given in claim 1, and pharmaceutically acceptable acid addition salts thereof and optionally additional therapeutically active compounds in admixture with suitable inert, non-toxic, pharmaceutical carriers and / or excipients.