FI70215B - FOERFARANDE FOER FRAMSTAELLNING AV NYA TERAPEUTISKT ANVAENDBARA APOVINKAMINOLDERIVAT OCH PHARMACEUTISKT GODTAGBARA SYRAADDITIONSSALTER DAERAV - Google Patents

Description

7021 5

The invention relates to a process for the preparation of novel therapeutically useful compounds of the formula I R1 10 rί 'Γη ^ ν 5 * 2 C_H_ (I), 0 ^ 3 CHo0 r 3 15 II

ch30 T

Wherein either R and R are both hydrogen or one of R 1 and R 2 is hydrogen and the other is a nitro group, and pharmaceutically acceptable acid addition salts thereof.

The process of the invention provides the preparation of apovincaminol 3 ', 4', 5'-trimethoxybenzoate of formula Ia or a pharmaceutically acceptable acid addition salt thereof / 25 Γ Γ [Π “ϊ! h 'U Jl A? .-of.

9-nitroapovincaminol-31,41,5'-trimethoxy-35 benzoate of the formula Ib or a pharmaceutically acceptable compound thereof, CH2- (Ia) 0 C2H5 CH-, acceptable acid addition salt, 2,7021 5 no2 f ^ Sj—

L Jl 1 H XL

5 jjA ^ Äs (Ib) ^ 2 C-Ης 0 2 5 ^ 3 °. c- 'fT Yo ίο

CH30 I

ch30 or 11-nitroapovincaminol-3 ', 4', 5'-trimethoxybenzoate of formula Ie or a pharmaceutically acceptable acid addition salt thereof

Ori0> * J33 20 O "2 C2H5 CH O C ^ (Ic) CH3O!

CH.JD

25

The process according to the invention for the preparation of the novel therapeutically useful apovincaminol derivatives of the formula I and their pharmaceutically acceptable acid addition salts is characterized in that apovinc-30 minol or its acid addition salt is reacted with 3,4,5-trimethoxybenzoic acid compound or convert it to a pharmaceutically acceptable acid addition salt or, if desired, nitrate a compound of formula 35a and separate the two components of the resulting mixture of compounds of formulas Ib and Ie and, if desired, convert the compound of formula Ib or Ie thus obtained to a pharmaceutically acceptable acid addition salt. salt in.

The process according to 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 3,4,5-trimethoxybenzoyl halide is used as the acylating agent, the reaction is carried out using an acid-binding agent in an equimolar amount or a small amount of more than 10. For this purpose, for example, an alkali metal carbonate, an alkali-carbonate carbonate or an organic amine can be used. If 3,4,5-trimethoxybenzoic acid is used as the acylating agent, the reaction is carried out using a catalytic amount of acid (a suitable acid is hydrochloric acid or sulfuric acid) or in the presence of a carboxyl group activating agent and / or a dehydrating agent. The carboxyl group can be activated with a halogenated phenol, more preferably pentachlorophenol. As the dehydrating agent, for example, N, N'-dicyclohexylcarbodiimide can be used. The acylation can be performed at a temperature between -20 ° C and the boiling point of the reaction mixture, most preferably at 20-60 ° C.

The product of formula Ia can be separated 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 an inorganic or organic acid (e.g. hydrochloric acid, sulfuric acid or phosphoric acid or tartaric acid, succinic acid, citric acid or ascorbic acid). The salt formation is carried out by traditionally known methods. It is preferable to add an acid solution in ethyl ether or acetone to the base solution. The salt formation takes place completely at a pH of 3-6.

The compound of formula Ia thus obtained can be nitrated if desired. The nitration can preferably be carried out with concentrated nitric acid. The most preferred reaction conditions are obtained using glacial acetic acid as the reaction medium. It is preferred to perform the nitration with cooling to about 0 ° C. When the reaction is complete, the excess acid is neutralized and the mixture of compounds of the formulas Ib and Ie is separated from the reaction mixture by extraction and / or evaporation.

5 The two components of the isomer mixture thus obtained can be separated. It is most preferred to carry out the separation of the isomers by chromatographic methods.

The compounds of formulas Ib and Ie may be converted into their pharmaceutically acceptable salts, if desired, by the methods described in connection with the preparation of the acid addition salts of the compound of formula Ia.

It is known that apovincaminol and its acetate have an effect on the coronary artery (FR patent 2,035,784). It is also known that apovincaminolbenzoo-15 accounts for general vasodilatory properties (HU Patent 166476; CA 83, 129279 v (1975)) and esters of apovinaminaminol formed with alkanecarboxylic acids having 3 to 12 carbon atoms show a vasodilatory effect on the brain. HU Patent 171662, 20 corresponding US Patent 4 108 996 and DE Patent 26 32 118).

Thus, all known esters of apovincaminol show vasodilatory effects. On the other hand, the novel compounds of this invention inhibit the enzyme activity of phosphodiesterase ent-25 and can be used primarily for the treatment of skin diseases involving the pathological rapid division of cells and the prevention of the recurrence of such diseases.

Diseases associated with rapid epidermal division are relatively common and occur in a few percent to 30 percent of the population. These diseases can be both benign and malignant in nature such as exfoliative disease, pruritic bending rash, allergic contact seizures, basal cell and germ cell cancer, fish scab disease, precursor of malignant keratosis, light-induced keratosis, pimple disease, and seborrheic disease. Some diseases occur only in humans, others in both humans and animals.

Il 5 70215

Because some skin diseases involving morbidly rapid proliferation of cells (e.g., dandruff disease) do not occur in animals, the effect of compounds against psoriasis can only be assessed indirectly from animal experiments.

5 Voorhess et al. (Arch. Derm. 104 (1971) 359-365) have found that the pathological rapid division of cells is associated with a decrease in the level of cyclic adenosine monophosphate (c-AMP). It is known that c-AMP is formed by adenyl cyclase and degraded by phosphodiesterase.

10 Voorhees was able to act on compounds with exfoliative disease that stimulate adenyl cyclase activity (such as norepinephrine) or inhibit phosphoric esterase activity (e.g., papaverine).

When designing the test as a model experiment, we proceeded from the 15 assumptions that Voorhees’s claim is also inversely true. Thus, if it can be shown that a particular compound inhibits phosphodiesterase activity, it can be implicitly assumed that said compound is suitable for the treatment of skin diseases involving the morbidly rapid division of cells.

This hypothesis proved to be correct: those compounds which had phosphodiesterase inhibitory activity in in vitro experiments also proved to be effective in the treatment of dandruff in clinical trials.

Our model experiment is performed with phosphorus esterase, which is isolated from animal body tissues (rat brain, bovine brain, bovine heart). The enzyme is isolated from J. Schröder and H.V. According to the method of Richenberg (Biochem. Biophys. Acta 302 (1973) 50), the isolated phosphodiesterase is purified by J.G. Hardman and E.W. Sutherland's method 30 (J. Biol. Chem. 240 (1965) 3704) and finally the activity of the purified enzyme is measured according to G. Pöch's radioisotope method for 10.1 millimoles of c-AMP substrate per excess of tritiated c-AMP. in the presence of 2.59 Bq Hc-AMP (in an incubation system, first without inhibitor and then in the presence of the apovincaminol-341,51-trimethoxybenzoate derivative as an inhibitor after 20 minutes of incubation (NS Arch. Pharmacol. 268 (1979) 272). A 1 millimolar stock solution of the test compound / 7021 5 is prepared and various amounts of said stock solution are added to the incubating enzyme preparation so that the concentration of the test compound in the incubated sample "7-6-6 is 5 x 10, 1 x 10, 5 x 10, -5 -5 - 4 5 1 x 10, 5 x 10 and 10 moles / liter An aqueous solution of the reference compound (papaverine) is added to an enzyme prepared in the same way.

The activity measured with the control (enzyme solution without inhibitor) is denoted as 100% and the activity of the solutions containing the compound of formula I 10 is expressed as a percentage of the control. The results measured with the enzyme isolated from rat brain are summarized in the following table.

Test compound (enzyme- Concentration of test compound in moles / liter of inhibitor) _, .6 Ί, .- 5 _, .- 5 _'_ 5 x 10_1 x 10_5 x 10 (Effect on enzyme activity,% of control)

Apovincaminol 3 ', 4', 5'-trimethoxybenzoate. HCl 64.5 49.2 47.2 20 (-) - 11-nitroapovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 53.6 44.4 37.3 (-) - 9-nitroapovincaminol-3 ', 4', 5'-trimethoxybenzoate. HCl 62.9 61.6 37.7 25 Papaverine 91.2 89.7 60.5

Results with an enzyme isolated from bovine brain and bovine heart are measured in the same manner. Using the results obtained, the enzyme activity is plotted per logarithm of the enzyme inhibitor concentration (expressed in μmol). The concentration of enzyme inhibitor that reduces enzyme activity by 50% (I ^ q) is read from the curve. The results obtained are summarized in the following table.

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Test compound 150 ~ values, in .moles by phosphorus esterase enzyme isolated from bovine heart from rat brain

Apovincaminol 3 ', 4', 5'-tri-methoxybenzoate. HCl 1,5 10 (-) - 11-nitroapovincaminol-3,4 ', 5'-trimethoxybenzoate · HCl 18 8 - (-) - 9-nitroapovincaminol-3', 4 ', 5'-trimethoxybenzoate · HCl 1 15

Papaverine · HCl 50 70

It can be seen from the above table that with the enzyme isolated from bovine heart and rat brain, the novel compounds of the present invention are 33-50 and 4.5-9, respectively, 15 times more active than papaverine used as a reference compound.

The first clinical trials were performed with topical compositions containing the active compound (ointment, cream, solution, tincture, paste, aerosol). Creams were used, each containing 2%, 1%, 0.5%, 0.25% and 0.1% of (-) - 9-nitro- or (-) - 11-nitroapovincaminol-3 ', 4', 51 -trimetoksibentsoaattia.

Patients with proriasis were treated. Another essential selection criterion was the fact that patients 25 did not receive concomitant systemic therapy for their underlying disease (i.e., immunosuppressive, cytostatic, or glucocorticoid therapy).

Groups of five patients were examined by the so-called spot method. One half of the symmetrically damaged skin area was treated with the active compound cream while the other half was treated with placebo. The patient's other damaged skin areas were treated with other local methods, e.g. an ointment commonly used for the treatment of dandruff and containing flumazone-35 pivalate and salicylic acid, said ointment being used as a reference.

____ Τ '~ 7021 5

The experiment was started with creams with a high concentration of the active substance, and other patients were treated with a cream with the lowest concentration of the active substance but still active. Treatment 5 was continued for two weeks twice daily without dressing.

The effect was assessed by observing three different symptoms - inflammation, cell proliferation and flaking. The intensity of symptoms was expressed on the following scale 0-3.

0 = no symptoms 2 = severe symptoms 10 1 = mild symptoms 3 = very severe symptoms

Symptoms were assessed before starting treatment (I), after seven days of treatment (II), and after fourteen days of treatment. The following table shows the mean scores (sum of scores divided by the number of patients).

An emulsion containing 2% of active compound was used.

Test compound Average number of points

Cell Accumulation Inflammation Exfoliation

_I II III I II III I II III

20

Compound of formula Ia 1.8 0.6 0.5 1.8 1.3 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 0

Compound of Formula Ie 2.4 1.8 1.3 2.2 1.3 1.3 1.6 1.20.8 25

The compounds according to the invention can be used in pharmaceutical compositions having phosphodiesterase inhibitory activity, which are mainly useful for the treatment of skin diseases involving the rapid proliferation of cells and for preventing the recurrence of such diseases, said compositions consisting of an active compound of formula I a compound or a pharmaceutically acceptable acid addition salt thereof and optionally other pharmaceutically active compounds in admixture with conventional pharmaceutical carriers and / or diluents.

9 70215

A suitable concentration of the active ingredient in the composition is 0.1-0.8%, preferably 0.2-2.0%. The compositions may optionally contain other pharmaceutically and medically active compounds such as antibiotics, cytosts, prostaglandins, ditranol, salicylic acid, tar, anti-inflammatory agents, immunosuppressants, glucocorticoids and, for parenteral administration, compositions suitable for parenteral administration. As a glucocorticoid, it is preferred to use triaminolone aceto-10-nide. It is preferable to prepare a composition containing the active compound suitable for topical use, such as creams, ointments, solutions, gels, sprays, foams, adhesives, etc.

The active ingredient is well suited for use in base-15 form, but acid addition salts may be used as well.

It is preferred to include the active ingredient in a cream which can be removed by washing.

Creams can be prepared by dissolving the active ingredient in an alcohol-type solvent, mainly propylene glycol or ethylene glycol or a mixture thereof with a small amount of water, and adding the resulting solution to an easily spreadable fat phase suitable for the skin.

Said fatty phase may be cetyl alcohol, stearyl alcohol, cetostearyl alcohol, paraffin oil, glycerol monostearate and the like.

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

The creams may preferably contain 0.25-2.0% of active ingredient, 45-50% of glycol, 23-27% of paraffin oil, 11-15% of stearyl alcohol and optionally up to 35 100% of other excipients.

The active compound can also be formulated in an ointment form which cannot be removed by washing with water, by incorporating the active compound directly into a fatty gas.

The active compound can also be formulated in solution or tincture form, which may contain e.g. 20-40% propylene glycol or dipropylene glycol, 40-55% 95% ethanol and up to 100% distilled water.

Spray compositions may be prepared by adding a fatty substance, e.g. myristate, and a propellant (e.g. freon) to a propylene glycol solution of the active compound.

Injectable solutions for parenteral administration, preferably subcutaneous or intradermal administration, may be prepared by dissolving the salt of the active compound in 0.72% aqueous sodium chloride solution and adjusting the pH of the solution to 5 ° C.

The pharmaceutical compositions may be prepared by methods known in the pharmaceutical art. It is possible to proceed by mixing the active compound and optionally other medically active compounds with suitable inert, non-toxic, known pharmaceutical carriers and / or excipients and formulating the mixture thus obtained in a pharmaceutically acceptable form.

The invention is illustrated in more detail by the following examples.

Example 1 Preparation of (-) - apovincaminol 3 ', 415'-trimethoxybenzoate 25.10 g (10.1 mmol) of (-) - apovincaminol are dissolved in 60 ml of anhydrous dichloromethane, followed by the addition of 3.10 g of anhydrous sodium carbonate and 2.50 g (10.9 mmol) of 3,4,5-trimethoxybenzoyl chloride-30 dia and the reaction mixture are stirred at room temperature for 24 hours, diluted with 100 ml of water, the organic phase is separated off and the aqueous phase is extracted twice with 20 ml of dichloromethane. The combined dichloromethane phases are dried over magnesium sulphate, filtered and the filtrate is evaporated to dryness in vacuo to give 4.50 g of the title product in a yield of 89.1%.

Il 7021 5

Molecular weight of the compound ^ 30 ^ 34 ^ 2 ^ 5 'IR 502.61 IR spectrum (as film): γ max 1725 cm -1 (-C = O); 1620 cm -1 (= C = O) 1 H-NMR spectrum (in deuterochloroform): <| : 1.01 (t, 3H, CH 3 CHO-); /: 3.72 (s, 6H, 2 -OCH 3); · {: 3.05 (s, 3H, --OCH3); Δ: 4.25 (s, 1H, ring prot.); Δ: 5.29 (s, 1H, --CH.dbd.); Δ: 5.4 (m, 2H, --OCH 2 -); d 6: 7.0-7.8 (m, 6H, aromatic). MS spectra: (m / e): 502 (53), 432 (100), 290 (17), 261 (41), 212 (18), 195 (35).

O c / jX, 7D = “2 2.0 ° (c = 0.7; dichloromethane) Example 2 Preparation of (-) - apovincaminol-3 ', 4', 5'-trimethoxybenzoate hydrogen tartrate

The compound prepared according to Example 1 is dissolved in diethyl ether. A saturated diethyl solution of D-tartaric acid is added to the solution until the hydrogen tartrate is completely precipitated.

The salt is filtered off and dried. Sp. 120-121 ° C.

IR spectrum (KBr): 1730 cm -1 (-C = O); 1640-1655 cm -1 (= C = C =) γ max K s / -X / p = -8.5 ° (c = 1; pyridine). Molecular weight 652.7. Example 3 Preparation of 20 - (-) - 9-nitroapovincaminol-31,41,51-trimethoxybenzoate and (-) - 11-nitroapovincaminol-3 ', 4', 5'-trimethoxybenzoate 5 g of (-) - apovincaminol Dissolve 31,4 ', 5'-trimethoxybenzoate in 50 ml of glacial acetic acid. A mixture of 20 ml of glacial acetic acid and 10 ml of concentrated nitric acid (d = 1.52) is added to the resulting solution at 25 ° C with stirring. The reaction mixture is poured into 350 ml of ice-cold water and the pH is adjusted to 9 by adding 25% aqueous ammonium hydroxide solution. The basic solution is extracted first with 250 ml and then twice with 200 ml of dichloromethane. The combined organic phases are dried over anhydrous sodium sulfate, filtered and the filtrate is evaporated to dryness under reduced pressure.

The isomer mixture thus obtained is separated into two components by chromatography on silica gel (Kie-35 selgel 60) as a 10: 2 mixture of benzene and acetone as adsorbent and eluent. Under these conditions, the R 2 value of the 9-nitro derivative 7021 5 12 is greater than the R 2 value of the 11-nitro derivative. There were thus obtained 1.7 g of the 9-nitro derivative and 1.8 g of the 11-nitro derivative.

The physical quantities of (_) - 9-nitroapovincaminol-31,41,5'-trimethoxybenzoate are as follows:

Sp. 77-78 ° C. [.alpha.] D @ 20 = -78.5 DEG (c = 1; chloroform). 1 H NMR Spectrum (CDCl 3): δ: 3.8 (s, 6H, 2 x 3.91 (s, 3H, 1 x CH 3 O-): 3.7 (d, 1H, H 12); .96 (d, 1H, ^η)? RV-: 8.1 (d, 1H, H 2 q).

The physical quantities of (-) - 11-nitroapovincaminol-31,4 ', 5'-trimethoxybenzoate are as follows:

Sp. 72-73 ° C. [Α] D = -134.3 ° (c = 1, chloroform). 1 H NMR Spectrum (CDCl 3): δ: 3.8 (s, 6H, 2 CH 3 O] F: 3.9 (S, 3H, 1 CH 3 O -); δ: 7.56 (d, 1H, H 9 ): 8.1 (d, 1H, H 2); λ / ": 8.9 (d, 1H, H 2).

Example 4 Preparation of 9-nitroapovincaminol-3 ', 41,51-trimethoxybenzoate hydrochloride and 11-nitroapovincaminol-3', 4 ', 5'-trimethoxybenzoate hydrochloride

9-Nitro- or II-nitroapovincaminol-3 ', 4', 5'-trimethoxybenzoate prepared according to Example 3 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 filtered off, washed and dried. The hydrogen chloride of the 9-nitro derivative melts at 144-150 ° C, while the hydrogen chloride of the 11-nitro derivative melts at 141-144 ° C.

Claims (8)

A process for the preparation of novel therapeutically useful compounds of formula I f1 f 'i; γ jL> X a .n ^ cA-4-J 10 .2 (I) „o C2H5 CH-.0. c '3 .. jl 15 ° ™ 3, 1 2 wherein either R and R are hydrogen or one of the symbols 1 2 R and R is hydrogen and the other a nitro group, and pharmaceutically acceptable acid addition salts thereof, reacting apovincaminol or an acid addition salt thereof with 3,4,5-trimethoxybenzoic acid or a reactive derivative thereof capable of effecting acylation, and isolating the compound of formula Ia in H1U.1 J n ^ CH2 Y'h (Ia) CH, 0 0 C2H5; o = Or convert it to a pharmaceutically acceptable acid addition salt, or, if desired, nitrate the compound of formula Ia, and separate the two components of the particular mixture of the compounds of formulas Ib and Ie, 7021 5 18 NO, i ^! 5 Ο-® * C2H5 «bl OijO CX 2 5 {'ί Ö 10 CH, 0 "" 2. A process according to claim 1, characterized in that apovine aminol-3 ', 4', 5'-trimethoxybenzoate of formula Ia or a pharmaceutically acceptable acid addition salt thereof is prepared. 30 L I J da) <y ^ 2 c2h5 CH, 0 C 'J' v, / II 35 \\ x | ° ca3o "{0CH3 7021 5 19 3. A process according to claim 1, characterized in that 9-nitroapovincinol-345'-trimethoxybenzoate of formula Ib or a pharmaceutically acceptable acid addition salt thereof, NO-Cnf "1 / U J. J ( Ib) 10 CH2 ^ CH o - 2 c2h5 CH-0 C 'J χγ "CH30" CH3O 3 in CH3i 15. CU .- ° ¾ cX, Ic> CH, 0 '° 25 3 / ° JO 0 ch30 ^ i And, if desired, converting a compound of formula Ib or Ic into a pharmaceutically acceptable acid addition salt. 4. A process according to claim 1, characterized in that 11-nitroapovinca-minol-3 ', 4', 51-trimethoxybenzoate of formula Ie or a pharmaceutically acceptable acid addition salt thereof is prepared. Ono ° 2 "" CH, "(IC>" .0 2 C2H5 25 CH-0 C 1 'M Π 1 0 CH30 "CH30 30 5. A process according to claim 1, characterized in that 3,4,5-trimethoxybenzoyl chloride is used as an acylating agent. 6. A process according to claim 1 or 5, characterized in that the acylation is carried out in the presence of an organic solvent.