JP2009502755A - Thiocolchicoside analogues with muscle relaxant and anti-inflammatory effects - Google Patents

Abstract

  The present invention relates to 3-demethoxy-3-aminothiocolchicine glycosyl derivatives of general formula (I), wherein R is hydrogen, methyl or hydroxymethyl. The compound of formula (I) has a muscle relaxing action and an anti-inflammatory action.

Description

  The present invention relates to a 3-demethoxy-3-aminothiocolchicine derivative having a muscle relaxing action and an anti-inflammatory action.

[Technical background]
Muscle relaxants share the property of reducing muscle tone and are commonly used to reduce pain due to tissue tension such as muscle contracture.

  Muscle contractures characterize some of the pathologies of motor organs and are one of the major factors in the persistence of pain associated with these pathologies. Muscle contractures are also inflammatory-rheumatic and occur in degenerative orthopedic conditions, and when they affect joints, they produce not only pain but also stiffness, mutual mobility of the joint ends, and thus Limit the functionality of the affected area. For these reasons, studies of molecules that confer muscle relaxant and non-contractive properties are still of considerable interest from a clinical point of view.

As is known, colchicine is a pseudoalkaloid that has been widely used in therapy for a long time for the treatment of gout. Thiocolchicoside known as (thiocolchicoside) 3- demethylthiocolchicine glucoside (C ₁₀ holds the thiomethyl group, and colchicine derivatives 3-position hydroxy groups are etherified by glucose molecules) are also inflammatory conditions muscle It is widely used in therapy for the treatment of skeletal contractures (Ortopedia e Traumatologia Oggi XII, n. 4. 1992). Recently, it has been demonstrated that the activity of thiocolchicosides can be attributed to the ability to interact with strychnine-sensitive glycine receptors, and therefore compounds with glycine-like activity have been treated with rheumatic orthopedic surgery due to their muscle relaxant properties. Can be used in the field.

[Disclosure of the Invention]
The present invention is directed to general formula (I):

Relates to a 3-demethoxy-3-aminothiocolchicine glycosyl derivative wherein R is hydrogen, methyl or hydroxymethyl.

  Thus, the pentose of 3-demethoxy-3-aminothiocolchicine and the α and β anomers of the D- and L-glycopyranosides of hexose are included in the compounds of formula (I).

  The compounds of the present invention generally have 3-demethoxy-3-aminothiocolchicine (II):

And formula (III):

Wherein X is OH, F, Cl, Br, I,
R ₂ is hydrogen, methyl, —CH ₂ —O—R ₃ ,
R ₃ is hydrogen or a hydroxy protecting group)
Prepared by reaction with a protected glycopyranoside.

  Of the radicals X, fluorine and bromine are preferred. Of the protecting groups, an acetyl group is preferred.

  As disclosed by Clark D et al. In WO 00/35865, 3-demethoxy-3-aminothiocolchicine (II) is prepared from 3-O-demethylthiocolchicine triflate, tosylate or mesylate by the Buchwald reaction.

  More particularly, 3-demethoxy-3-aminothiocolchicine is reacted with an appropriate glycoside and optionally protected with a hydroxy residue. The reaction is carried out at a temperature ranging from 25 ° C. to the boiling point of the solvent, preferably in a polar protic solvent selected from alcohols (MeOH). The reaction is usually complete in a time period ranging from 8 hours to 6 days. If necessary, the hydrolysis of the protecting group can also be carried out directly on the crude reaction product without recovering the intermediate.

  When the reaction is carried out using electromagnetic waves, a significant decrease in reaction time and an increase in yield are observed. More particularly, 3-demethoxy-3-aminothiocolchicine is carried out at 110 ° C. and 150 watts in a polar protic solvent selected from suitable glycosides and preferably alcohols (eg MeOH). The reaction is usually complete after 3 hours.

In order to determine the affinity of the resulting product for the strychnine-sensitive binding site of the murine spinal cord, a displacement test was measured in vitro using ^[3H] strikinine as the ligand. The results suggest that the derivative behaves as an allosteric compound on spinal strychnine-sensitive glycine receptors. This interaction is a sign of muscle relaxant action of the compounds of the invention (Cimino M. et al., Eur. J. Pharmacol. 1996, 318. 201-204; Balduini W. et al., Neuropharmacol. 2001, 40. 1044-1049).

  It has been found that the compounds of formula (II) are active and therefore the invention also relates to the use of the compounds for the preparation of a medicament having a muscle relaxant action.

  The compounds of the present invention can be formulated by conventional methods, together with excipients, as a formulation intended for oral, intravenous, intramuscular, transdermal, topical administration (eg, Remington's Pharmaceutical Sciences). Handbook, XVII Ed., Mack Pub., NY, USA). Naturally and synthetic phospholipids are particularly preferred as excipients useful for the preparation of liposome forms for parenteral or topical administration. The dose can range from 5 to 50 mg depending on the disease and route of administration.

  The invention will be explained in more detail by means of several examples.

[Experiment part]
Melting points were measured using a Buchi 510 instrument. NMR spectra were obtained on a Bruker AC 500 instrument. IR spectra were obtained with a Jasco IR Report 100 spectrophotometer. The reaction was carried out in a Milstone Micro SYNTH microwave oven. [Α] ₂₅ ^D values were measured with a 343-PLUS Perkin-Helmer polarimeter.

Example 1.3-O-trifluoromethanesulfonyl -3-O-demethyl-thio coli heat Chin nitrogen atmosphere, at 0 ° C., 3-O-demethylthiocolchicine in anhydrous _{CH 2 Cl 2 (50ml) (} 2g, 4 To a solution of .98 mmol) and p-DMAP (1.77 g, 15.78 mmol) was added trifluoromethanesulfonic anhydride (1.24 ml, 7.40 mmol).

Stirring was continued at 0 ° C. for 20 hours and then at room temperature for 3 hours. The reaction was monitored by TLC analysis (10: 1, CH ₂ Cl ₂ / MeOH, Rf: initial value = 0.27, product = 0.38). After evaporation of the solvent, the residue was purified by column chromatography on alumina using a CH ₂ Cl ₂ / MeOH mixture (increasing polarity gradient). The product was obtained as a yellow solid after crystallization from ethanol (1.84 g, 70%):

Example 2.3- demethoxy-3-under-amino thiocolchicine nitrogen _{atmosphere, Cs 2 CO 3 (685mg,} 2.09mmol), Pd (OAc) 2 (68mg, 0.29mmol), (±) BINAP (290mg, 0.44 mmol) and the compound obtained in Example 1 (800 mg, 1.48 mmol) were introduced into a Schlenk round bottom flask. The mixture was heated to 40 ° C. under reduced pressure for 2 hours. After cooling, properly degassed anhydrous toluene (3 ml) was added followed by benzophenone imine (0.25 ml, 1.49 mmol). The round bottom flask was sealed with a rubber stopper and filled with nitrogen. The reaction mixture was stirred for 45 minutes at room temperature, then heated to 120 ° C. for 16 hours and monitored by TLC analysis (20: 1, CH ₂ Cl ₂ / MeOH). The mixture was cooled to room temperature, diluted with AcOEt, filtered and concentrated under reduced pressure. The crude product containing the desired imino adduct was dissolved in MeOH (15 ml) at room temperature. To the solution was added AcONa (509 mg, 6.21 mmol) and _{NH 2 OH · HCl (323mg,} 4.65mmol). After 30 minutes, the mixture was diluted with CH ₂ Cl ₂ (5 ml) and acidified to pH 2 with 2N HCl. The organic phase was separated and the aqueous phase was alkalized with 25% NaOH solution and extracted with CH ₂ Cl ₂ (3 × 20 ml). The organic phase is dried over Na ₂ SO ₄ and concentrated under pressure to give the desired product (TLC, 10: 1.5, CH ₂ Cl _{2 /} MeOH, Rf 0.43), which is then Crystallized from EtOH (373 mg, 60%):

Example 3. General procedure for the synthesis of 3-demethoxy-3-aminothiocolchicine glucoside

Method A
To a solution of 3-demethoxy-3-aminothiocolchicine (404 mg, 1 mmol) in MeOH (8.5 ml) was added commercially available glycopyranoside (1 mmol) and the mixture was heated at 80 ° C. for 6 days in a sealed tube. did. The reaction was monitored by TLC analysis (10: 1.5 CH ₂ Cl ₂ / MeOH). When the solvent was distilled off under pressure, TLC analysis of the crude reaction product showed the presence of the starting reagent (8-10%) and an isomer mixture of aminoglycosides. This was separated by column chromatography on silica gel (100: 1 to 100: 5 CH ₂ Cl ₂ / MeOH) and then crystallized from MeOH / iPr ₂ O to give the desired compound.

Method B
To a solution of 3-O-demethoxy-3-aminothiocolchicine (404 mg, 1 mmol) in MeOH (8.5 ml) was added commercially available glycopyranoside (1 mmol) and the mixture was placed in an electromagnetic oven for 4 hours at 110 ° C. And heated at 150 watts. The reaction was monitored by TLC analysis (10: 1.5 CH ₂ Cl ₂ / MeOH). The solvent was distilled off under pressure and TLC analysis of the reaction crude product showed the presence of starting reagent and an isomeric mixture of aminoglycosides. These were separated by column chromatography on silica gel (100: 1 to 100: 5 CH ₂ Cl ₂ / MeOH) and then recrystallized from MeOH / iPr ₂ O to give the desired compound.

  The table below lists the yield and diastereoselectivity of the process.

Example 4. 3-N-α-D-glucopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 5. 3-N-β-D-glucopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 6. 3-N-α-D-arabinopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 7. 3-ND-arabinopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 8. 3-N-α-D-Loxopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 9. 3-N-β-D-lyxopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 10.3-N-α-D-xylopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 11.3-N-β-D-xylopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 12. 3-N-α-L-rhamnopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 13. 3-N-β-D-mannopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Example 14. 3-N-β-L-fucopyranosyl-3-O-demethoxy-3-aminothiocolchicine

Claims (5)

Formula (I):

(Where
R is hydrogen, methyl or hydroxymethyl). 3-ND-xylopyranosyl-3-O-demethoxy-3-aminothiocolchicine;
3-ND-glucopyranosyl-3-O-demethoxy-3-aminothiocolchicine;
3-ND-mannopyranosyl-3-O-demethoxy-3-aminothiocolchicine;
3-ND-arabinopyranosyl-3-O-demethoxy-3-aminothiocolchicine;
3-ND-lyxopyranosyl-3-O-demethoxy-3-aminothiocolchicine;
3-NL-rhamnopyranosyl-3-O-demethoxy-3-aminothiocolchicine;
A compound selected from 3-N-L-fucopyranosyl-3-O-demethoxy-3-aminothiocolchicine.   A pharmaceutical composition comprising the compound of claim 1 or 2 mixed with an acceptable carrier and / or excipient.   Use of the compound according to claim 1 or 2 for the manufacture of a medicament having muscle relaxant action and anti-inflammatory action. Formula (II) for the manufacture of a medicament having muscle relaxant action:

Use of the compound.