EP2167067A1

EP2167067A1 - Use of eflucimibe for the production of a medicament for the prevention or treatment of a disease caused by sebaceous gland dysfunction in humans or animals

Info

Publication number: EP2167067A1
Application number: EP08826957A
Authority: EP
Inventors: Thibaud Biadatti-Portal
Original assignee: Galderma Research and Development SNC
Current assignee: Galderma Research and Development SNC
Priority date: 2007-07-19
Filing date: 2008-07-17
Publication date: 2010-03-31
Also published as: FR2918889B1; US20100172957A1; FR2918889A1; CA2692837A1; WO2009019385A1

Abstract

The invention relates to the use of at least one of the compounds (S)-2-dodecylsulfanyl-N-(4-hydroxy-2,3,5-trimethyl-phenyl)-2-phenyl-acetanilide (eflucimibe) of formula (I), (R)-2-dodecylsulfanyl-N-(4-hydroxy-2,3,5-trimethyl-phenyl)-2-phenyl-acetanilide ((R) enantiomer), and the pharmaceutically acceptable salts and solvates thereof for producing a medicament or a dermatological composition for preventing and treating disorders caused by sebaceous gland dysfunction and/or any state or disease related to excess sebum production in humans or animals.

Description

Use of eflucimibe for the preparation of a medicament for preventing or treating a disease caused by dysfunction of the sebaceous glands in humans or animals

The present invention relates to a new use of eflucimibe, or a pharmaceutically acceptable salt thereof, pharmaceutically acceptable solvates or hydrates, for the preparation of a medicament for preventing or treating a disorder due to dysfunction of the sebaceous glands. in humans or animals, such as acne and / or any state or pathology related to overproduction of sebum, such as seborrheic dermatitis, oily skin or oily scalp.

The patent application WO 97/19918 describes a family of 2,3,5-trimethyl-4-hydroxy anilide derivatives which are inhibitors of the enzyme ACAT (Acyl CoA: cholesterol O-Acyl transferase). These compounds are presented in this patent application as hypocholesterolemic and antioxidant compounds, which can be used for the treatment of hypercholesterolemia or atherosclerosis. Inhibition of the ACAT enzyme blocks the esterification of free cholesterol to cholesterol esters. Moreover, this enzyme is expressed in the sebaceous glands. The sebaceous glands are holocrine glands that secrete a mixture of lipids known as sebum. Cholesterol esters account for 2-3% of the lipids in human sebum. The excess of sebum is conducive to the appearance of acne or other dermatological pathologies, and in any case gives the skin a greasy and unattractive appearance. Current treatments to reduce excess sebum are unsatisfactory. By way of example, mention may be made of isotretinoin, which, although decreasing by nearly 90% the production of sebum, has significant side effects. No effective ACAT inhibitor treatment has yet been developed.

It has now been demonstrated that among the inhibitors of the ACAT enzyme described in WO 97/19918, (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2 -phenylacetanilide, known as eflucimibe, of formula (I): optionally in the form of a salt, a solvate and / or a pharmaceutically acceptable hydrate, may be used to prevent or treat disorders resulting from dysfunction of the sebaceous glands in humans or animals and / or any state or pathology related to overproduction of sebum. In the context of the invention, the compound (I) will preferably be used in its pure enantiomeric (S) form as represented, but may also be used in its enantiomeric (R) form or in the form of a mixture (racemic ) of these two compounds, in any proportion.

Thus, the subject of the present invention is the use of at least one compound chosen from (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenyl- acetanilide (eflucimibe) of formula (I):

(R) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide (enantiomer (R)), pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, for the preparation of a medicament for preventing or treating disorders due to dysfunction of the sebaceous glands in humans or animals or any condition or condition related to overproduction of sebum.

The subject of the invention is also a product chosen from (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide

(eflucimibe) of formula (I): (R) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide (enantiomer (R)), pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, for its use in the treatment or prevention of a disorder due to a dysfunction of the sebaceous glands in humans or animals and / or a condition or pathology related to overproduction of sebum.

Preferably, the product chosen from (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide (eflucimibe) of formula (I):

(R) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide (enantiomer (R)), their pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, are used in the treatment or prevention of acne, seborrheic dermatitis, oily skin or oily scalp.

Eflucimibe is described in Example 7 of WO 97/19918. The pharmaceutically acceptable salts of the compound of formula (I) include those with organic or inorganic bases, for example salts of alkali or alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium salts. The solvates of eflucimibe, of its enantiomer (R), or of their salts, represent the hydrates of such compounds and / or the combination of such compounds with a linear or branched C ₁ -C ₄ alcohol, such as methanol, ethanol, isopropanol, or n-propanol.

In the context of the invention, the inventors have demonstrated that eflucimibe makes it possible to fight against disorders due to dysfunction of the sebaceous glands in humans or animals and / or any state or pathology related to overproduction. of sebum. Disorders due to dysfunction of the sebaceous glands are preferably acne.

As a condition or pathology related to an overproduction of sebum is meant, in particular, seborrheic dermatitis, oily skin or oily scalp.

For its use as a drug, eflucimibe, its enantiomer or a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, must be formulated into a pharmaceutical composition, preferably dermatological.

The present invention therefore also relates to pharmaceutical compositions, preferably dermatological, comprising, in a physiologically acceptable carrier, at least one compound selected from eflucimibe, the enantiomer (R) of eflucimibe, their pharmaceutically acceptable salts, and pharmaceutically acceptable solvates thereof for the treatment and / or prevention of disorders due to sebaceous gland dysfunction and / or any condition or condition related to overproduction of sebum. By physiologically acceptable support is meant a medium compatible with the skin, mucous membranes and / or integuments.

Eflucimibe or its enantiomeric form (R), optionally in pharmaceutically acceptable salt form, or pharmaceutically acceptable solvate or hydrate may also be used for the manufacture of pharmaceutical compositions, preferably dermatological for treating oily skin or fatty scalps. Such compositions may be intended and therefore suitable for oral, topical, enteral, parenteral, ocular, sublingual, inhaled, subcutaneous, intramuscular, intravenous, transdermal, local or rectal.

Oral and topical forms are nevertheless preferred. Eflucimibe or its enantiomeric form (R) optionally in the form of a salt or a pharmaceutically acceptable solvate, alone or in combination with another active ingredient, may be administered in unit dosage form, in admixture with conventional pharmaceutical carriers or excipients, to animals and humans. Preferably, the pharmaceutical composition is packaged in a form suitable for oral or topical application. Topically, we mean an application on the skin and / or the mucous membranes.

The compositions according to the invention comprise eflucimibe, or its enantiomeric form (R), or one of its pharmaceutically acceptable salts, or pharmaceutically acceptable solvates, in an amount sufficient to obtain the desired prophylactic or therapeutic effect. The dosage varies with the age, sex and weight of the patient. The compound (I), or its enantiomeric form (R) or a salt thereof, or solvates will preferably be administered at a rate of 0.01 to 100 mg / kg and per day, advantageously from 0.01 to 50 mg / kg and per day. It is also possible to administer such doses in 2 to 4 daily administrations. Although these assays are examples of average situations, there may be special cases where higher or lower dosages are appropriate, such assays also belong to the invention.

The compositions according to the invention comprise a physiologically acceptable carrier chosen according to the desired pharmaceutical and preferably dermatological form and the chosen mode of administration. Said support comprises at least one pharmaceutically acceptable excipient.

For oral administration, the pharmaceutical composition, preferably dermatological, may be in the form of tablets, capsules, dragees, pills, syrups, suspensions, solutions, powders, granules, emulsions, capsules, microspheres or nanospheres or lipid or polymeric vesicles allowing controlled release. Parenterally, the composition may be in the form of solutions or suspensions for infusion or injection. To obtain a solid composition for oral administration, the active ingredient may be mixed with at least one inert diluent, such as sucrose, lactose or starch. In general, other additives, such as a lubricant such as magnesium stearate, may be added. In the case of capsules, tablets or pills in particular, a buffer may be added. In the case of oral liquid compositions, an inert diluent such as water may be used.

Topically, the pharmaceutical composition according to the invention is more particularly intended for the treatment of skin and mucous membranes and may be in the form of ointments, creams, milks, ointments, powders, impregnated swabs, syndets, solutions, gels, sprays, mousses, suspensions, lotions, sticks, shampoos, or washing bases. It may also be in the form of suspensions of microspheres or nanospheres or lipid or polymeric vesicles or polymeric patches and hydrogels allowing controlled release. This topical composition may be in anhydrous form, in aqueous form or in the form of an emulsion.

Eflucimibe (I), or its enantiomeric form (R) or a salt or solvate thereof, when administered orally is administered at a rate of 0.01 to 100 mg / kg and per day, preferably 0 to , 01 to 50 mg / kg and per day.

Eflucimibe (I), or its enantiomeric form (R) or one of its salts or solvates, when administered topically is used at a concentration generally of between 0.001 and 10% by weight, preferably between 0, 01 and 5% by weight, relative to the total weight of the composition.

It is also possible, for the treatment of disorders due to dysfunctions of the sebaceous glands, eflucimibe, or its enantiomeric form

(R) or a pharmaceutically acceptable salt or a pharmaceutically acceptable solvates or hydrates thereof, in combination with another active ingredient.

The pharmaceutical compositions, preferably dermatological as described above, may therefore contain inert or even pharmacodynamically active additives, or combinations of these additives, and in particular: - wetting agents;

- flavor enhancers;

preserving agents such as esters of parahydroxybenzoic acid;

stabilizing agents; humidity regulating agents;

pH regulating agents;

osmotic pressure modifying agents;

emulsifying agents;

UV-A and UV-B filters; - emollients;

moisturizing agents such as glycerol, PEG 400, thiamorpholinone and its derivatives or urea.

Of course, those skilled in the art will take care to choose the optional compound (s) to be added to these compositions in such a way that the effect on the disorders due to dysfunctions of the desired sebaceous glands is not, or not substantially, impaired by the addition envisaged.

The compositions below are given for illustrative purposes:

Example 1: COMPOSITIONS

A- ORAL WAY

Tablet of 0.2 α

- Eflucimibe 0.001 g

- 0.1 14 g starch

- Dicalcium phosphate 0.020 g

- Silica 0.020 g

- Lactose 0.030 g

- Talc 0.010 g - Magnesium stearate 0.005 g

B- TOPICAL ROUTE (a) Ointment

- Eflucimibe 0.30 g - White vaseline codex qs 100 g (b) Lotion

- Eflucimibe 0.10 g

- Polyethylene glycol (PEG 400) 69.90 g - 95% Ethanol 30.00 g

The study of the properties of eflucimibe has shown that eflucimibe, as well as its enantiomer (R), its salts, or pharmaceutically acceptable solvates are not toxic, and allow to reduce sebum production by the sebaceous glands.

EXAMPLE 2 BIOLOGICAL RESULTS

Measure of ACAT / SOAT activity

The purpose of this test is to visualize the ACAT / SOAT enzyme activity by measuring the synthesis of cholesterol esters.

The test is inspired from the following publication: "Identification of ACAT1- and ACAT2- specifies inhibitors using a novel, cell-based fluorescence assay: individual ACAT uniqueness," J. Lipid.Res (2004) vol 45, pages 378-386.

The principle of this test is based on the use of NBD-Cholesterol, a cholesterol analogue whose fluorescence depends on its environment. When it is in a polar environment, it is weakly fluorescent whereas in a non-polar environment, it is strongly fluorescent. NBD-free Cholesterol is localized in cell membranes and is weakly fluorescent in this polar environment. When NBD-cholesterol is esterified with ACAT, the NBD-cholesterol ester is localized in the non-polar lipid droplets and is then strongly fluorescent.

Method:

The HepG2 cells are incubated in the presence of NBD-cholesterol (1 μg / ml) and the product to be tested in 96-well black plates with a transparent bottom at the rate of 30,000 cells per well. After incubation for 6 h at 37 ° C, 5% CO2 medium is removed by inversion and the cells are washed with 2 times 100 μl of PBS. After addition of 50 .mu.l of lysis buffer (10 mM NaPO.sub.4, 1% Igepal), the plates are stirred for 5 min and read in fluorescence (490 nm excitation, 540 nm emission) on the FUSION (Perkin Elmer).

Eflucimibe in this test potently inhibits the synthesis of cholesterol esters with an IC ₅₀ of 4 nM.

Evaluation of the activity of eflucimibe on the size of the sebaceous glands in the female Fuzzy rat by topical administration

The purpose of the study inspired by the publication of Uno and Kunata, 1993, JID, 101, 143S-147S. Fang Ye et al. 1997, is to evaluate the ability of eflucimibe to alter the size of sebaceous glands in female Fuzzy rats.

Method:

After daily topical administration of a volume of 100 μl_ of test product or vehicle for 25 days in the appropriate vehicle, the size of the sebaceous glands on epidermal split is evaluated.

6 animals per group are treated with daily oral administrations with a volume of 10ml / kg of test product or vehicle.

8mm biopsies are taken from each animal to evaluate the size of the epidermal split sebaceous glands.

After a three-day incubation period in 4 DMEM medium, the biopsies are transferred to a 12-well plate each containing 1 ml of 1M sodium bromide for 2 hours at 37 ° C. The dermis is delicately separated from the epidermis and an image is acquired by a binocular magnifying glass and then analyzed using the TINA software to identify the size of the sebaceous glands.

The results showed that the reduction of 50% in the size of the sebaceous glands is obtained at a concentration of 0.00089% of the compound according to the invention. This value of EC50 demonstrates a very strong activity of the eflucimibe.

Inhibition of cholesterol ester synthesis in rat preputial sebocytes in primary culture Rat preputial gland sebocytes are a recognized model for studying the activity of compounds on sebum production and lipid synthesis in sebocytes. The general principle is based on the collection of preputial glands, the separation and culture of sebocytes, and the treatment of these cell cultures by the test compounds. These molecules are supplemented with radioactive substrates that must be included in the lipid composition of sebum (acetate, fatty acids, waxy alcohols or cholesterol labeled with Carbon 14). An analysis of the radioactivity contained in the supernatant after cell lysis, by thin layer chromatography and reading of the radioactivity indicates the loss of radioactivity after treatment with the test compounds and thus the reduction of lipid synthesis.

The glands are removed sterilely and put in a solution of DMEM + antibiotics 1% + Vancomycin 100μg / ml. A first wash with PBS is performed, then the connective tissue of all glands is removed before a second wash. The cut glands are diluted in a solution of collagenase 1 mg / ml at the rate of 1 gland / ml of solution. The culture is stirred in 37 ° water bath for 15 minutes. After treatment with trypsin then a sequence of centrifugations-samples, the cells are diluted in a volume of complete DMEM so as to obtain a concentration equal to 1 .106 cells / ml. The obtained sebocytes are seeded in the wells in the presence of 3 t3 mitomycin cells thawed extemporaneously and the plates are incubated at 37 ± 5% CO 2 for 3 days.

At D + 3 after culturing, the medium is removed. The cells are rinsed with PBS. PBS is removed and the serum free Cellgro medium is added along with the solutions of test compounds in DMSO. At D + 6, the Cellgro medium is again added together with the test molecules under the same conditions as at D + 3.

At D + 8-9, the 14C-acetate radiolabelled substrate (equivalent of 2 μCi / well) is added and the medium is incubated between 18 and 24 h at 37 ° + 5% CO 2, then the medium is removed, the cells rinsed with PBS. (-). Lysis of the cells is carried out with a trypsin-EDTA mixture, incubation for 30 min at 37 °. The contents of each well are taken and a 2/1 dichloromethane / methanol mixture is added, and the samples are then deposited on a silica plate using a pipettor-depositor automaton.

Elution: 1st heptane migration 9cm, 2nd migration toluene 8.5cm, 3rd migration mix 5.75cm (heptane, ethyl ether, acetic acid 70/30/1). The image analyzer after drying takes place on a STORM phosphorimager associated with TINA software to analyze the image.

In this model, eflucimibe inhibits the synthesis of cholesterol esters in rat preputial sebocytes with 83% inhibition and an IC50 of 625 nM.

Claims

Use of at least one compound chosen from (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethyl-phenyl) -2-phenyl-acetanilide of formula (I):

(R) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide, pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, for the preparation of a medicament intended to prevent or treat a disorder due to dysfunction of the sebaceous glands in humans or animals and / or any state or pathology related to overproduction of sebum.

2. Use according to claim 1 characterized in that the disorder due to dysfunction of the sebaceous glands in humans or animals is acne.

3. Use according to claim 1 characterized in that the condition or pathology related to overproduction of sebum in humans or animals is seborrheic dermatitis, oily skin or oily scalp.

4. Use according to one of claims 1 to 3 characterized in that the drug is suitable for oral administration.

5. Use according to one of claims 1 to 3 characterized in that the drug is adapted for topical administration.

6 - Use according to one of claims 1 to 5, characterized in that the salts of (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethyl-phenyl) -2-phenyl- acetanilide or (R) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethylphenyl) -2-phenylacetanilide are the salts of these compounds with alkali or alkaline earth metals.

7. Use according to one of claims 1 to 5, characterized in that the solvates of (S) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethyl-phenyl) -2-phenyl- acetanilide or (R) -2-dodecylsulfanyl-N- (4-hydroxy-2,3,5-trimethyl-phenyl) -2-phenyl-acetanilide are the hydrates of these compounds.

8. Use according to claim 5 characterized in that the medicament is in the form of an ointment, cream, milk, ointments, powder, swab soaked, syndet, solution, gel, spray, foam, suspension, lotion, stick, shampoo, or washing base.