EP1648915A1

EP1648915A1 - Steroid derivatives and use thereof as medicaments

Info

Publication number: EP1648915A1
Application number: EP04767779A
Authority: EP
Inventors: Gilles Gutierrez; Mostafa Serrar; Zohra Hadid
Original assignee: Inovat; INOVAT Sarl
Current assignee: Inovat; INOVAT Sarl
Priority date: 2003-07-25
Filing date: 2004-07-23
Publication date: 2006-04-26
Also published as: US20060258753A1; US7186756B2; FR2857968B1; WO2005014614A1; CA2536486A1; JP2006528652A; FR2857968A1

Abstract

The invention concerns the field of chemistry and, more particularly, the field of organic chemistry. The invention specifically relates to novel A-nor steroids according to general formula (I) in which R represents hydrogen, a lower alkyl radical having 1 to 7 carbon atoms, a lower cycloalkyl radical having 3 to 7 carbon atoms, an aryl radical having 5 to 10 carbon atoms, an arylalkyl radical in which the alkyl group has 1 to 6 carbon atoms, or an acyl radical derived from an aliphatic carboxylic acid, from a cycloalkyl carboxylic acid or from an aryl carboxylic acid. The invention also relates to pharmaceutical compositions containing, as an active substance, at least one compound of general formula (I) in association or mixed with an excipient or an appropriate vehicle. The inventive pharmaceutical compositions can also contain another active substance that having a similar or synergetic action. The compounds of general formula (I) are used as medicaments that aid in fixing calcium by osteoblasts.

Description

STEROID DERIVATIVES AND THEIR USE AS MEDICINES

The present invention relates to the field of chemistry and more particularly to that of organic chemistry. More particularly, it relates to new steroids of semi-synthetic origin belonging to the A-nor Steroids family. It specifically relates to new steroids corresponding to the general formula I

(I)

where R is:

- hydrogen

- a lower alkyl radical having from 1 to 7 carbon atoms,

- a lower cycloalkyl radical having from 3 to 7 carbon atoms,

- an aryl radical having 5 to 10 carbon atoms,

- an arylalkyl radical in which the alkyl group has from 1 to 6 carbon atoms,

- or an acyl radical derived from an aliphatic carboxylic acid, a cycloalkylcarboxylic acid or an arylcarboxylic acid. Its chemical name is 2-oxo 3-OR A-nor 25- ethylidene Cholest 3-ene. In addition, the keto-enol structure of this molecule allows the formation of complexes such as for example the formation of a complex with Iron, Copper or Zinc. The compound of formula I for which R is hydrogen, may exist in one of the tautomeric forms of the keto-enol form, depending on the medium and according to the pH. The ketone form can become more or less completely enolized to result in either a ketone / enol mixture or a completely enolized enol + ketone compound. An alkaline medium or the use of polar solvents are the factors favorable to enolization and complete enolization of the molecule makes it possible to obtain a pure and stable 2-keto 3-enol compound. Among the compounds of general formula I, there may be mentioned very particularly: 2-oxo 3-hydroxy 25-ethylidene A-nor cholest-3ene (R = H), - 2-oxo 3-acetoxy 25-ethylidene A-nor cholest -3ene (R = CH ₃ CO), 2-oxo 3-methoxy 25-ethylidene A-nor cholest-3ene (R = CH ₃ ) which are the preferred compounds. It is also possible to prepare a tetrahydropyran ether by the action of dihydrofuran in an acid medium, a benzyl ether by the action of benzyl chloride or nitrobenzyl chloride in the presence of dimethylformamide. It is also possible to prepare various carboxylic esters such as a propionate, a valerate, a benzoate, a naphthoate, a terephthalate, a succinate, a malonate, a nicotinate, a glucuronate, or a lactobionate. The compounds of general formula I can be produced by hemisynthesis from fucosterol according to a process in which an oxidation of fucosterol to cholestene-5 3-one is carried out followed by oxidation by DDQ to cholestestene-5-2,3 dione and rearrangement in 2-keto 3-hydroxy A-nor cholestene-3 in an alkaline medium. The formation of ethers from the 3-hydroxylated compound is carried out by means of a diazoalkane in an inert solvent such as isopropyl ether or tetrahydrofuran, or by the action of an alkylating agent in basic medium. The alkylating agent is preferably a halide, a sulfate, an alkyl tosylate, a cycloalkyl halide, an arylalkyl or aryl halide. The procedure is carried out in the presence of pyridine, lutidine, collidine, dimethylformamide, dimethylacetamide, or else in the presence of 4-dimethylaminopyridine. The esters of the compound of formula I can be prepared by the action of an acylating agent, such as a functional derivative of organic carboxylic acid, an acid halide, an acid anhydride, or a mixed anhydride of acid, in a polar aprotic solvent in the presence of an acylation catalyst such as 4-dimethylaminopyridine or 4-hydroxybenzotriazole on the 3-hydroxylated compound. The structure of the compounds of formula I was demonstrated by determining the NMR spectrum and, in particular, the NMR spectrum with a high resolution device (500 MHz) (NMR of the proton [Η], of carbon ¹³ C, in HMBC, NOE, TOCSY and NMR-LC). Additional analysis by proton NMR [ ^X H] shows that the product, having spent ten days in cold CD3OD, has not degraded. The compound of formula I, for which R is hydrogen, can give rise to metal complexes with metals of valence II, III or IV such as the complexes of Iron, Nickel, Copper, Zinc, Manganese or Chrome. They can serve as a mode of identification, purification or separation of the compounds of formula I during physical analyzes. The compounds of general formula I also give rise to derivatives of the free ketone function such as, for example, a ketal, a thioketal, a hemithioketal, an oxime, an 0-carboxymethyloxime, or an optically active or racemic (dicarboxyalkylene ketal). general I have seven centers of asymmetry and can therefore exist in different spatial structures, so that the junction of cycles B and C can have the natural configuration 9β-δα or the antipodal configuration 9α-8β depending on the synthesis conditions. The configuration of methyl in position 20 on the side chain is in principle β. This orientation can, if desired, be reversed. The compounds of formula I are also defined by the nature of the UV spectrum which exhibits a strong absorption at 220 and 240 nm. The addition of acid does not modify the peaks of UV absorption.

Alkaline products like sodium carbonate, potash or lithine cause a shift from 240 nm to 255 nm. The compounds according to the invention can also be characterized by other methods of analysis such as circular dichroism, the infrared spectrum in the dry state, or dispersed in Nujol, thin layer chromatography or chromatography. high performance in liquid phase, or rotational power in ethanol. The compounds of general formula I are conveniently called Maltadiolone and their esters. We will call the molecule Maltadiolone

(I) the maltan skeleton being

The compounds of general formula I show interesting biological properties which make them useful as active principles of medicament. The substances according to the invention exert an influence on the synthesis of connective molecules such as desomosial proteins and cytokeratins of the skin even in the presence of corrosive agents such as sulfuric acid. The compounds of general formula I are distinguished by remarkable repair properties of the extracellular matrix, they increase the synthesis of collagen, they promote the synthesis of glycosaminoglycans, even in the presence of deleterious substances such as Interleukins and mainly Interleukin IL - 1. For example, a culture of bone cells (line

UMR 106 or G 292) treated with 10 ng of Maltadiolone shows an increase in fixed calcium compared to the untreated control cells. For example, a culture of bone cells (UMR line 106 or G 292) will be treated with 5 μl dlnterleukin-1, (IL-1), ie 1 ng per ml. Analysis by atomic spectrophometric of the calcium fixed in the extracellular matrix shows a rate of 700 ng / ml in the control cells. This value is lowered to 25 ng / ml in the presence of dlL-1. Under the same conditions, the cells are treated with 10 μg of Verapamil or any other calcium channel blocker such as Cinchonine or Diltiazem. It is found that the quantity of calcium fixed in the extracellular matrix of the osteoblasts, is equivalent to that of the cells treated with lnterleukin. Under the same conditions, the cells treated both with 1 ng dlL-1 and with Maltadiolone 10 μg, according to the invention, restore a calcium-binding activity equivalent to that of cells not treated with 11L-1. Under the same conditions, cells treated with both 10 μg of Verapamil or Diltiazem and 10 μg of Maltadiolone have a calcium-binding activity equivalent to that of cells not treated with a calcium channel blocker. Under the same conditions as above, it can be seen that the activity of restoring the fixation of calcium on the extracellular matrix is perceptible from 10 ng / ml. The invention also relates to pharmaceutical compositions containing, as active principle, at least one compound of general formula I, in which R has the meanings provided above, in combination or in mixture with an excipient or a vehicle suitable for administration by digestive, parenteral, rectal or topical, inert, non-toxic, pharmaceutically acceptable route. For digestive administration, the compounds of general formula I are in the form of naked or coated tablets, dragees, pills, flavored or non-flavored powders, capsules, capsules. For parenteral administration, the compounds of general formula I are packaged in the form of injectable solutions, injectable suspensions, injectable dispersions, in a water in oil or oil in water emulsion. A particularly vehicle suitable is an emulsion of medium chain fatty acids, marketed under the name intralipid. The pharmaceutical compositions according to the invention may also contain another active principle of similar or synergistic calciotropic action such as an estrogenic product such as for example estradiol, an estradiol ester, or an estradiol ether such as Metranol or Quingestanol, a SERM. The compounds of general formula I are distinguished by a high level of activity at low doses for very low toxicity. The normal dosage of compounds of general formula I ranges from 10 ng to 50 mcg / ml and preferably from 50 ng to 500 ng / ml. The phenomena of toxicity appear at doses greater than 50 mcg / ml and rather indicate an overactivity of the products. The following examples illustrate the invention without, however, limiting it.

Example I Formulation of 2-oxo 3-hydroxy 25-ethylidene A-nor Cholest 3-ene Tablets

Active ingredient 100 μg Lactose 100 g Microcrystalline cellulose 25 g Polyvinylpyprolidone K 30 5 g Sorbitol 10 g Calcium sulphate 25 g per 1,000 tablets Example II Formulation of 2-oxo 3-hydroxy 25-ethylidene A-nor Cholest 3-ene tablets

Active ingredient 2.25 mg Mannitol 5 g Sugar syrup 10 g Magnesium carbonate 500 g Talc 15 g

Compression on STOKES rotary machines to make 70 mg tablets.

Example III Formulation of 2-oxo 3-Methoxy 25-ethylidene A-nor Cholest 3-ene Tablets

Active ingredient 250 μg Calcium carbonate 300 g Microcrystalline cellulose 20 g Ethylene oxide polymer (PLURONIC F 88) 20 g Colloidal silica 15 g

Per 1,000 tablets

Example IV Solution for injection of 2-oxo-3-acetoxy-25-ethylidene A- nor Cholest 3-ene 2-oxo 3-acetoxy 25-ethylidene A- nor Cholest 3-ene 5 mg Intralipid 250 ml Distilled water 500 ml

The solution is filled in 2 ml ampoules. It is filtered through a membrane and sterilized at 120 ° C for ten minutes. The bulbs are distributed in rigid boxes pointed at the bottom. By applying a vacuum, the ampoules are filled, they are wiped, dried and the tips are welded in a sterile atmosphere. Example V 2-oxo-3-hydroxy 25-ethylidene A-nor Cholest 3-ene ferrous complex tablets. 0.82 g of 2-oxo 3-hydroxy 25-ethylene A-nor Cholest 3-ene is dissolved in 25 ml of dimethylformamide. This solution is diluted with an equal volume of water and then added without delay

10 ml of a 5% solution of ferrous sulfate in dilute sulfuric acid. It is mixed vigorously and then left to stand overnight. The precipitate formed is separated on a sintered glass crucible, rinsed several times with water and then twice consecutively with acetone. 1.02 g of ferrous complex are thus collected and dried at 80 ° C. in a ventilated oven. After an overnight stay, the complex, which is in the form of dark brown crystals, is collected. Weigh 0.100 g of ferrous complex and 500 g of lactose.

One homogenizes carefully then one adds 40 g of microcrystalline cellulose then 25 g of colloidal silica (Aerosil 200) and, finally, 4 g of Magnesium stearate. The powder, after homogenization, is sieved through a 200 sieve. 27 mg capsules containing 4 mcg of active principle per unit dose are prepared with this powder.

Claims

R E V E N D I C A T I O N S 1 °) New steroids corresponding to the general formula I

(I)

in which R is hydrogen, a lower alkyl radical having from 1 to 7 carbon atoms, a lower cycloalkyl radical having from 3 to 7 carbon atoms, an aryl radical having from 5 to 10 carbon atoms, an arylalkyl radical where the alkyl group has from 1 to 6 carbon atoms, or an acyl radical derived from an aliphatic carboxylic acid, a cycloalkoylcarboxylic acid or an arylcarboxylic acid.

2 °) Tautomeric forms of the new steroids according to claim 1.

3) metal complexes of the compound according to claim 1 for which R is hydrogen.

4 °) steroid blocking compounds of general formula I according to claim 1, in which the ketone function is blocked in the form of a ketal, thioketal, hemithioketal, oxime or optically active ketal or (dicarboxyalcoylene) or racemic 5 °) A compound according to one of the preceding claims, namely 2-oxo-3-hydroxy 25-ethylidene A-nor Cholest-3-ene

6 °) A compound according to claim 1, namely 2-oxo-3-acetoxy 25-ethylidene A-nor cholest-3-ene

7 °) A compound according to claim 1, namely 2-oxo 3-methoxy 25-ethylidene A-nor Cholest-3-th.

8 °) Process for the preparation of the compounds of general formula I according to one of the preceding claims, in which R is an alkyl, aralkyl, cycloalkyl or aryl radical, characterized in that the 3-hydroxylated derivative is subjected to the action of 'a diazoalkane in an inert solvent of an alkyl, cycloalkyl, aryl or arylalkyl halide, sulfate or tosylate.

9 °) Process for the preparation of the compounds of general formula I according to one of the preceding claims, in which R is an acyl radical derived from an aliphatic carboxylic, cycloal ylcarboxylic, arylcarboxylic or arylalkylcarboxylic acid in which the compound is subjected to which R is hydrogen to the action of an acylating agent in a polar aprotic solvent in the presence of an acylation catalyst.

10 °) Pharmaceutical compositions containing as active principle at least one compound of general formula I in association or in mixture with an excipient or an inert, non-toxic vehicle which is therapeutically compatible.

11 °) Pharmaceutical compositions according to claim 10, in which the excipient or vehicle is one of those suitable for administration by the digestive, parenteral, rectal or topical route.

12 °) Pharmaceutical compositions according to claim 10 or claim 11, in which the content of compound of general formula I ranges from 50 ng to 20 mcg per unit dose.

13 °) Pharmaceutical compositions according to claim 12, in which the content of compound of general formula I ranges from 50 ng to 500 ng per unit dosage.

14 °) Pharmaceutical compositions according to one of claims 10 to 13, in which another active ingredient of similar or synergistic action is introduced.