BG98439A - Quaternary ammonium derivatives of (-) and (+) 10-(1-azobicyclo /2.2.2/oct-3-il-methyl)-1oh-phenothiazine, pharmaceutical compositions on their basis and method for their preparation - Google Patents

Abstract

The invention relates to a dextrorotatory (+) and levorotatory (-) enantiomorphic form of the 10- (1-azabicyclo [2.2.2] oct-3-ylmethyl) -10n- phenothiazine quaternary ammonium derivatives of the general formula is C1-4 alkyl or cyclopropyl, x is halogen, dimethyl sulphate or p-toluenesulfonic acid, a process for their preparation, as well as pharmaceutical compositions containing as an active ingredient and for use in the treatment of bronchospastic and allergic diseases, in particular of bronchial asthma and rhinitis.

Description

The invention relates to right-handed (+> and left-handed (-) enantiomorphic Forms of quaternary ammonium derivatives of 10- (1-azobicyclo C2.2.23 oct-3-yl-methyl) -10H-Phenothiazine with the following general Formula (I)

where

R = C1- a, alkyl, cyclopropyl,

X = halogen, dimethyl sulphate, p-tolu uffonic acid, to the method aa thereof, and to their containing

Pharmaceutical compositions which are useful in the treatment of bronchospastic and allergic diseases, in particular bronchial asthma and rhinitis.

2.

BACKGROUND OF THE INVENTION

From the literature known 10- (1-Azobicyclic Phenothiazine (internationally the following Formula (A), e.g., FR-A-2034605, is

E2.2.23 oct-3-yl-methyl) -10H Known as mecitazine with

as well as its antihistamine properties described therein.

In EP-A-159059, the Applicant describes the quaternary derivatives of the compound of Formula (A) in the Form of Racemic Modifications, having appreciable anti-birch-elastic activity.

SUMMARY OF THE INVENTION

The quaternary derivatives of mecitazine, as well as mecitazine itself, have an asymmetric carbon atom (position 3 on 1 — the azobicycle E2.2.23 octane). Each racemic quaternary derivative thus consists of two (+) and (-) enantiomorphic Forms in the ratio of 50:50.

quaternary derivatives of

It was surprisingly found,

the enantiomorphic forms of

with Formula (I)

X where

R = C1- + alkyl or cyclopropyl,

X = halogen, dimethyl sulfate or p-toluenesulfonic acid exhibit different properties, which makes each of them particularly suitable for different treatments, respectively.

According to the present invention C 1-3 alkyl means: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl.

According to the present invention halogen means: iodine, bromine, chlorine.

In particular, it has been found that while both enantiomorphic Forms according to the general Formula (I) are equally active with acetylcholine, each has surprisingly different properties, for example the right-handed (+) enantiomorphic Form is significantly more effective with histamine. than the corresponding left-spinning (-) enantiomorphic Form. In addition, the right-handed enantiomorphic form was found to be less toxic than the left-handed one.

The right-handed (+) enantiomorphic Form can therefore be administered according to the present invention as an agent particularly suited for the treatment of bronchospastic and allergic diseases, especially bronchial asthma and rhinitis, as well as all histamine-related pathologies, one of the main mediators asthmatic bronchial spasms, and abnormal response of the nasal mucosa; or with acetylcholine - which controls both the tone of the respiratory system and the mucous membrane of the nose. As is known, the mediators leading to asthmatic bronchial spasms and rhinitis are many, and the increase in their impact depends on the type and extent of the disease, as well as on the patient. The fact that the right-handed enantiomorphic Form influences the two mediators when in fairly significant concentrations makes this Form extremely interesting as a tool for treating the relevant diseases.

For the preparation of the enantiomorphic Forms according to the invention, the racemic mixture of mecitazine (Formula A) is separated into two corresponding right- and left-handed enantiomorphic Forms by the phase chromatographic separation technique. In particular, cellulose chromatographic columns may be used, and as eluent respectively hexane / ethyl alcohol (94/6), hexane methyl alcohol (98/2), hexane / ethyl alcohol / TEA (98 / 0.8 / 1.2). The enantiomorphic Forms obtained are then quaternary derivatives by the reagents described in the examples given hereinafter by their indications, which do not limit the invention.

The enantiomorphic Formulations prepared according to the invention are particularly useful for the preparation of Pharmaceutical compositions for external use (mainly by inhalation or by smearing) or for oral administration. One of the Pharmaceutical compositions suitable for administration based on an active ingredient according to the invention is disclosed in Example 3 in a manner that does not limit the invention.

Examples of carrying out the invention

Example 1.

(-) - 1-methyl-3- (1 (-H-phenothiazine-10-yl-methyl) -1-azobicyclo C2.2.2J octane iodide according to Formula (I) at R = CH ₃ and X = I ^- .

g (0.1 mol) of mecitazine (-) enantiomer The form was obtained in a chromatographic chiral-phase column from the racemic mixture of mecitazine, which was suspended in 200 ml of acetonitrile. The suspension was stirred for 10 minutes until homogenized. Add 50 ml (0.2 mol) of methyl iodide. The solution thus obtained is heated to 50 ° C, with heating at this temperature for 16 hours. The reaction mixture was then cooled to 0-4 ° C afterwards, holding the mixture at trays for 48 hours. The resulting solid crystals were filtered under vacuum, washed on the filter with ethyl ether, and then dried under vacuum at SO ^ C to constant weight.

Obtained a final product weighing 42 g (yield 9O'X)

5.

under the form of solid crystals of cream color and the following

Physico-chemical characteristics:

150 x 4.6 column with HPLC Nucleozil

Eluent: MeOH / CH ₃ CN / NH 2 H ₃ PO 4 = 43/25/32;

Flow rate (flow rate): 0.5 ml / min, = 254 nm;

Injection: 4 ^ in methyl alcohol;

Overall peak at r.t .: 6.4;

Melting point: 142-144 ° C;

Ct /] D ^2O s -21.3 ° C (1% conc. In OH).

Elemental analysis of C ₃ iH _3lb IN ₌ S:

p N. I N S calculated 54.31 5.59 27.35 6.03 6.89 found 54.06 5.60 27.20 5.94 6.80

1HNMR (CDCl3): 7.21-6.84 (2m, 8H, 11 ', 2 *, 3', 4 ', 6', 7 ', 8', 9'1); 4.27-3.93 (m, 2H, 191); 3.93-3.46 (m, 6H 12,6,71);

3.21 (s, 3H, C101); 2.71 (m, 1H, 131); 2.30 (D, 1H 141); 1.99-1.77 (2t, 4H, 15.81)

1SC NMR (CDCl ₃ ): 144.62 (C-9'a, C-10 * a); 127.77 <C-1 ', C-4',

C-6 ', C-9'); 126.97 (C-4'a, C-5a); 123.26 (C-2 *, C-8 ');

116.25 (C-3 ', C-7'); 60.49 (C-9); 57.38 (C-6); 57.21 (C-7);

52.54 (C-3); 48.30 (C-2); 32.06 (C-10); 24.87 (C-8)

20.90 (C-4); 19.57 (C-5).

The optical purity product was chromatographically analyzed in the free phase by chiral-phase HPLC column, for example CHIRACEL O.D.® eluent column - hexane / ethyl alcohol = 94/6, flow rate 0.5 ml / min. = 254 nm, 4 ^ injection into hexane / ethyl alcohol, peak - 98X at rt: 15.03.

Example 2.

(+) -1-methyl-3- (1—-H-Phenothiavin-10-yl-methyl) -1-azobicyclo 12.2.21 octane iodide according to Formula (I) at R = CH 3 and X = I '.

g (0.08 mol) of mecitazine (+) enantiomorphic Form according to the invention is obtained by separation by chromatographic chiral-phase column of the racemic mixture of mecitazine suspended in 160 ml of acetonitrile.

6.

The suspension was stirred for 10 min. until homogenized. 40 ml (0.64 mol) of methyl iodide are added. The resulting solution was heated to 5O &lt; 0 &gt; C, held at this temperature for 16 hours. The reaction mixture was then cooled to 20 &lt; 0 &gt; O, then to 0-4 &lt; 0 &gt; C and stored at this temperature for hours. The resulting solid crystals were filtered off under vacuum, washed on the filter with ethyl ether and dried under vacuum at 50 ° C to constant weight.

35 g of the desired product (947 yield) are obtained in the form of cream-colored solid crystals having the following physicochemical characteristics:

NMR; NMR was as in Example 1.

HPLC was as in Example 1, with the chiral phase HPLC peaking at r.t. - 16.73 (977), the melting point was 142-145 ° C, CcilD20: + 21ο (1Ζ conc in CH 2 OH).

Elemental analysis for C21H2 & .IN2S

P N. I N S calculated 54.31 5.59 27.35 6.03 6.89 found 54.00 5.62 27. 18 5.96 6.82

Example 3.

In vitro tests on a pharmaceutical composition consisting of: active ingredient - E (+) or (-) product

according to Formula (1) 3 - 100 mg; sorbitan trioleate (Arlacel 85 ^j) - 20 mg; Freon 11 ^it - 3.380 mg; Freon 12-114 ^R (65:35) - 10,500 mg.

The above composition is placed in a small cylinder with a metering valve.

The composition prepared as above was divided into unit doses containing from 50 to 200 / hg of active ingredient.

The composition thus prepared is further tested with:

Activity tests

The enantiomorphic Forms prepared according to Example 1 and Example 2 were tested and compared for their pharmacotoxicological properties and were specifically tested for the following aspects: acetylcholine

Ί.

and histamine antagonism on guinea pig isolated from guinea pigs; retention of acetylcholine- and histamine-induced bronchial contraction in guinea pigs (by Konzett-Roessler method), acute toxicity in mice.

The results obtained indicate that both enantiomorphic Forms are comparable antagonists of acetylcholine and histamine in guinea pig trachea. While both enantiomorphic Forms are equally effective for acetylcholine (pA = 7.96 and 7.81 aa for the left-rotating and for the right-rotating enantiomorphic Forms), the (+) right-rotating enantiomorphic Form (pAa for 8.05) is more reversible (pAa for 8.05). of histamine. The results of the in vitro assays are supported by the results of the in vivo assays, which demonstrate that the two enantiomorphic Forms are equally effective in inhibiting acetylcholine-induced bronchial contraction. At the same time, the spinning (+) enantio-morphine form is significantly more effective than the spinning (+) enzyme in inhibiting histamine-induced bronchial contraction (ED50: 63 and 239 nmol kg-1, respectively)

Toxicity Test

The acute toxicity test for iv administration to mice showed that the right-handed (+) enantiomorphic Form was significantly less toxic ELDoo and the base determined at 95% was 3.65 (3.45-3-86) mg kg ^_1 l compared to left-handed (-) enantiomorphic Form E2.68 (2.61

2.74) mg kg ~ ^x 3.

The test results are given in the following

Table 1:

8.

In vitro tests

Determination of antagonism of

TABLE 1 acetylcholine and histamine on guinea pig trachea isolated

Acetylcholine

Histamine pA

Left-handed (-> enantiomorphic

Form (Example 1)

7.96

7.09

Right-spinning (+) enantiomorphic

Form (Example 2)

7.81

8.05

Acetylcholine

Histamine

ED ₌ o (nmol kg “ ¹ )

Left-handed (-) enantiomorphic

Form (Example 1)

Right-spinning (+) enantiomorphic

Form (Example 2)

Acute toxicity in mice

LD50 (mg kg * and base (base) determined at 95 “Z)

Left-handed (-) enantiomorphic

Form (Example 1)

2.68 (2.61-2.74)

Right-spinning (+) enantiomorphic

Form (Example 2)

86)

Claims (9)

Patent Claims 1. Enantiomorphic Formula 10- (1-Azobicyclo [2.2.23 cct3-yl-methyl) -HO-Phenothiazine quaternary ammonium derivatives of the following general Formula CI) wherein R is C 1-6 alkyl or cyclopropyl; X is halogen, dimethyl sulfate or p-toluenesulfonate. acid. 2. Enantiomorphic (+) Form of 10- (1-azobicyclo [2.2.21] oct3-yl-methyl) -HO-Phenothiazine Quaternary ammonium derivatives of the following general Formula (I) 3 ' 2 * β ® (I) X 10. where R is C 1-6 alkyl or CINLOPROPYL X is halogen, dimethyl sulfate or p-toluenesulfonic acid. 3. Enantiomorphic Form according to claim 1 and —Y where R e dream, and I "1 1" 4. Enantiomorphic Form according to claim 1 and 'Ί Λ. , where R e CH's, · H · C1. 5. Enantiomorphic Form according to claim 1 and X-, where R e CH3 :, and I · Br, 6. Enantiomorphic Form according to claim 1 and 7-¾ X-, where R e CH3, and K is dimethyl sulfate. 7. Enantiomorphic Form according to claim 1 and Ο χ>, where R e CH: c, and X is p-toluenesulfonic acid. 8. Method for the preparation of (-) or (+) enantiomorphic Forms of 10- (1-Azobicyclo C2.2.23 oct-3 ~ yl-methyl) -10H ~ Phenothiazine quaternary ammonium derivatives of general Formula (I) wherein R is C 1-6 alkyl or cycloalkyl and X is halogen, dimethylsulfate or p-toluenesulfonate an acid characterized in that the racemic mixture of mecitazine of Formula (A) is separated into two right and left-handed enantiomorphic Forms by chromatographic separation technique using chiral Phases, then quaternary derivatives are obtained with the corresponding reagents. A pharmaceutical composition comprising, as an active ingredient, a 'levorotatory (-) enantiomorphic Form according to claim 1.3 to 7. A pharmaceutical composition comprising as an active ingredient a dextrorotatory (+) enantiomorphic Form according to claim 2 to 7. A method for preparing a pharmaceutical composition according to claim * 9 and 10, characterized in that a pharmaceutically effective amount of (+) or (-) 10- (1-azobicyclo C2.2.21 oct-3-yl-methyl> -10H -Phenothiazine quaternary ammonium derivative of Formula (I) 3 ' 2 * c ® (I) ι *? X X- where R is C1-, alkyl or cyclopropyl, X is halogen, dimethyl sulfate or p-toluenesulfonic acid mixed with suitable carriers to formulate a composition suitable for E = external or oral use. Use of the composition of claim 9 for the treatment of bronchial asthma and rhinitis. The composition of claim 1 for the treatment of bronchial asthma and rhinitis.