WO2015114194A1

WO2015114194A1 - Liposomed sulphur composition

Info

Publication number: WO2015114194A1
Application number: PCT/ES2015/070062
Authority: WO
Inventors: Pedro González Enseñat
Original assignee: Enoc Solutions, S. L.
Priority date: 2014-01-29
Filing date: 2015-01-29
Publication date: 2015-08-06
Also published as: ES2542088B1; ES2542088A1

Abstract

The invention concerns a composition comprising a suspension of liposomes which contain elemental sulphur incorporated in the lipid membrane thereof. The invention also relates to the use of this composition in the treatment of skin diseases, such as seborrheic dermatitis or acne.

Description

COMPOSITION OF LIPOSOMATE SULFUR DESCRIPTION The present invention relates to a composition of liposomes containing sulfur element incorporated into its lipid membrane and the use of said composition in the treatment of diseases, mainly dermatological, such as seborrheic dermatitis or acne, in addition to others. . Therefore, the invention could be framed in the field of pharmacology and cosmetics.

STATE OF THE TECHNIQUE

The use of sulfur as a therapeutic agent for various dermatological and non-dermatological disorders or diseases dates from time immemorial. For many years, element sulfur has been considered topically in different forms or galenic preparations as the specific remedy of seborrhea. Due to their low solubility or their solubility in solvents not allowed or not suitable for human or animal use, the activity of said formulations has always been relative, low or low and therefore the preparations have had to incorporate high concentrations of said element to compensate for this problem, between 2% and 10% or 20% of it. Said low activity or efficiency has always been related and determined or conditioned by its suspension / dilution form and the particle size of the sulfur element incorporated in the formulation, both direct consequences of its low or almost zero solubility.

The topical applications of the element sulfur in different forms or galenic preparations are multiple. Due to its keratolytic effect, it is useful in the treatment and prevention of seborrheic dermatitis, acne, rosacea, pityriasis versicolor, some parasitosis such as scabiosis, folliculitis and other skin diseases such as some superficial dermatophytosis, perioral dermatitis, necrotic acne, and certain forms of eczema and psoriasis. It has also been used at low concentrations as keratoplastic, as an antipruritic, and, in addition, it has oxidoreductive, vasomotor, photosensitizing, "depigmenting" activity, certain germicidal and fungicidal activity. At high doses it can be irritating, produce itching / burning, reddening of the skin, exfoliation or hyperproliferation. In general, the element sulfur has been used in the treatment of some intestinal parasitosis, also as a trace element because of its "nutritive" character essential for the synthesis of many amino acids, for hair loss, strengthening nails, etc.

Of all its therapeutic possibilities, the topical dermatological applications mentioned are the most frequent and relevant so far.

The most historically known topical element sulfur formulations or forms are precipitated sulfur and sublimed sulfur. Both formulations or forms contain element sulfur with a minimum purity grade of 99.5% in anhydrous form. The galenic and therapeutic experience with the element sulfur, sublimed or precipitated, has led experts to conclude that the smaller the particle size of the element sulfur in the formulation, the greater and more homogeneous the degree of interaction or contact with the area to be treated and, therefore, greater will also be its therapeutic efficacy.

In order to improve its galenic and functional characteristics, other sulfur preparations such as colloidal sulfur, "bioazufre", sulfurized potash, etc. have been developed. Other examples of this are polythionic acids, their alkaline salts, quaternary ammonium polythionates, and mercaptocarboxylic acids, either in the form of acid, esters or amides. Interesting functional results have been obtained using some of the mineral or organic sulfur derivatives, whether natural or synthetic, although generally their organoleptic characteristics, especially the bad smell they give off, complicate or condition their use and acceptance.

In addition, the galenic problem of the element sulfur and other forms thereof includes in many cases the need to use certain components as solvents or dispersants, such as fats or oils, for their final formulation, with an occlusive and greasing effect, the application of which may be counterproductive. In the pathology to be treated, an example of this happens with this type of formulation and pathologies such as seborrheic dermatitis, acne, etc. The therapeutic sulfur is currently applied to the skin and legs in very different forms or galenic preparations, such as ointments, creams, gels, lotions or milks, soaps or shampoos, masks and ointments. Its application at high doses and concentrations can be irritating, produce peeling, burning, reddening of the skin, exfoliation, hyperproliferation ... so there is a need for a sulfur delivery system that is effective and does not require high concentrations of this to minimize possible adverse reactions, facilitate its application and improve its organoleptic characteristics.

DESCRIPTION OF THE INVENTION The incorporation of the element sulfur in liposomes with a good functional activity, in a galenically correct formulation, and, organoleptically very acceptable is the innovation developed and object of this patent. This encapsulation of the sulfur element in liposomes has been carried out in order to solubilize, suspend and / or disperse it without the need to use non-permitted solvents or other non-convenient products, thereby solving the main galenic problem thereof, as well as its Once its functional aspect also improves considerably, because, due to its nanometric size and the physicochemical characteristics of said liposomes, once applied, the sulfur element in liposomal form will be transported by said nanostructures, improving its pharmacokinetic and pharmacodynamic characteristics. In addition to the problems of solubility, galenic formulation, and functional sulfur element, the resulting product, in its liposomal form, is organoleptically much better, easier to apply and tolerable or pleasing to patients, both for its smell and its possible textures and for its ease of application or use, all in comparison with the conventional or conventional forms thereof.

Just as the concentrations of element sulfur used in conventional forms are high, between 20 and 200 mg / g, in the liposomal form they range between 0.02 and 0.4 mg / ml.

The final product, a water-based liposomal suspension, is very easy and quick to apply, such as atomization, rapid absorption and has no occlusive or greasing effect on the treated area. It can be easily applied to faneras, hairs and nails, with the same advantages. Just stains the skin or clothes. In the liposomal form of the invention, the efficacy of the element sulfur in the treatment or prevention of certain pathologies is enhanced, not only by the galenic, pharmacokinetic and pharmacodynamic contributions of the liposomal structure, but also by the recognized anti-seborrheic-anticomedogenic activity of phosphatidylcholine in particular, and unsaturated phospholipids in general, main components of the liposomal membrane, and that act synergistically with sulfur in these pathologies. The antiseborrheic-anticomedogenic efficacy of these compounds is mainly attributed to their high content of linoleic acid.

The advantages of the present invention are listed below:

- The liposome when solubilizing, suspending or dispersing the element sulfur in an aqueous phase in the form of tiny and very homogeneous particles offers a very good galenic alternative to said active ingredient.

- The liposome due to its special physicochemical characteristics improves the therapeutic properties of the element sulfur compared to the classical or conventional galenic forms. This improvement is due to the special pharmacodynamic and pharmacokinetic characteristics of the sulfur element encapsulated in the liposomes.

- The improvements or advantages mentioned above allow the use of lower concentrations and doses of the asset while maintaining its therapeutic activity, reducing the risk of adverse or unwanted reactions, such as irritation, burning ...

- Pharmaceutical formulations that incorporate the liposomal form of sulfur element have better organoleptic characteristics, especially odor, compared to those that contain it in their concentrations and conventional galenic forms.

- The product in liposomal, water-based form, in addition to being applied and absorbed much more easily than conventional formulations, especially fats, does not grease and occlude the skin, leaves no color on it, and hardly stains clothing.

- The special and very versatile characteristics of the liposomal formulation of sulfur element allow it to be incorporated and presented in many galenic forms, both in fluid form, lotion, suspension, serum, atomizer, ... as well as in semi-solid forms such as gels, emulsions , emulgeles, ointments, masks, etc. - The liposomal "concentrate" or the liposomes with sulfur element is / are easily incorporated / so associable / s to other formulations or products that in addition to it contain other or other synergistic or complementary assets intended for the treatment of different pathologies in order to improve their effectiveness and / or tolerance.

- The galenic, pharmacokinetic and pharmacodynamic improvements provided by the liposomes of the invention in general, because of its great biocompatibility can be administered by any route improving and enhancing the effects of the element sulfur, allow us to think about new applications of the same by any route of administration and for pathologies not currently contemplated as capable of being treated with sulfur element.

- The antiseborrheic-anticomedogenic effect of phosphatidylcholine, the main component of the liposomal membrane, which acts synergistically with the element sulfur in certain pathologies, also means an important advantage of the liposomal form of the same compared to conventional ones.

- The increase in bactericidal capacity of liposomes containing element sulfur, compared to element sulfur in its conventional form, means an improvement in its activity and has the advantage of reinforcing the antiacneic, anti-medogenic, anti-rosacea effect, etc. of the product.

- The physicochemical characteristics, the ease and versatility of the formulation of said liposomal element sulfur form, have the advantage of not needing products that interfere or may be counterproductive for the treatment of certain pathologies, or for the patient's own health. For example, you do not need fats, oils or other solvents or components for its formulation.

- The preparation has characteristics, particle size, which allow it to be sterilized both in its concentrate phase and also in its diluted form, as a finished product. This possibility implies the advantage of allowing the products to be packaged and administered without the need to add preservatives to the formulation.

Therefore, a first aspect of the present invention relates to a composition comprising an aqueous phase liposome suspension characterized in that it comprises:

Phospholipids with a phosphatidylcholine ratio between 75 and 99.5% by weight,

^■ at least one bile salt,

^■ at least one alcohol and

^■ water,

and because liposomes contain sulfur element incorporated into the phospholipid bilayer. The phospholipids of the composition may come from natural lecithins extracted or be of synthetic origin. The term "liposome" means a spherical vesicle with at least one membrane composed of a double layer of phospholipids, also called phospholipid bilayer, which consist of hydrophilic and lipophilic parts.

The liposomes can be single, oligo- or multilamellar, that is, they can comprise one or more double layers of phospholipids. Although those of the invention, the most suitable and preferred for topical application are unilamellar.

By "composition comprising liposomes" is meant compositions that give rise to the formation of liposomes, whether uni-, oligo- or multilamellar and micellar solutions convertible by dilution in liposomes, although preferably unilamellar liposomes are of interest.

The term "sulfur element incorporated into the bilayer" means that sulfur is part or is included in the lipid bilayer structure of the liposome. The combination of the components of the liposome and the sulfur element as an active principle takes place during their preparation and prior to the manufacture of the liposomal suspension. The term "sulfur element" means any form of mineral sulfur with a minimum of 99.5% purity, although preferably the substance known as "flower sulfur" is used which, having an adequate grain size and characteristics and therapeutic and pharmaceutically recognized properties, it becomes the active of said liposomal formulation or preparation.

The encapsulation of sulfur element in liposomes leads to a good alternative or solution of the difficulties of its galenic formulation, to an improvement of the functional aspects or characteristics thereof, and, consequently, to a significant reduction of the necessary dose thereof with the same or superior therapeutic efficacy, compared to conventional or non-conventional liposomal forms. The disadvantages of its formulation and presentation, as well as a large part of its possible Side effects are drastically reduced or even disappear. Some examples of this general improvement are better galenic characteristics of the product, especially, an improvement of its organoleptic characteristics, greater ease of formulation, greater ease and convenience of application, greater safety and safety margin of the product, less possibility of overdosing and the occurrence of adverse reactions.

In a preferred embodiment the element sulfur liposomes are first produced or manufactured in a concentrated form that can be stored for long periods of time, and / or, secondly, be diluted or incorporated into another formulation at a certain concentration for application. in patients In certain cases the product can be applied in the concentrated form of the product, although it must be done very carefully. In a preferred embodiment of a final product for application in patients, the composition is presented or prepared in the form of an aqueous suspension at a concentration of sulfur element in liposomal form appropriate or suitable for direct topical application in the form of lotion, atomized or with eyedropper, associated or not with other components or assets.

In another preferred embodiment, the composition is presented in a form suitable for topical, oral, intravenous, intramuscular, intraperitoneal, subcutaneous, cutaneous, nail, capillary or inhalation topical administration. Preferably by topical cutaneous, nail and capillary route (skin and pimples).

In another preferred embodiment, the liposomes have an average diameter between 40 nm and 500 nm, preferably between 60 nm and 180 nm, and more preferably between 75 nm and 145 nm. In another preferred embodiment of the invention, the sulfur element incorporated in the liposomes is pharmaceutical grade sulfur flower. Preferably in the final product for the patient the concentration of this element is between 0.004 gr per liter to 0.4 gr per liter, more preferably between 0.012 gr per liter and 0.25 gr per liter, and even more preferably between 0.02 gr per liter and 0.12 gr per liter. In the concentrate resulting from the manufacture of the flower sulfur liposomes typical of invention, the sulfur is preferably at a concentration of 0.4% and in the final product for the patient the preferred concentration is 0.04%.

In another preferred embodiment of the invention, the phospholipid concentration is from 10 g to 200 g per liter of composition, more preferably between 50 g and 150 g per liter of composition and even more preferably between 90 g and 10 g per liter of composition. In the concentrate resulting from the manufacture of the flower sulfur liposomes of the invention, the phospholipid concentration is preferably 100 g / liter and in the final product is 10 g / liter.

In another preferred embodiment, the phospholipids have the formula (I):

OR

II

R1 - C -0- CH2

I

R ₂ - C -0- CH O

YES II

O CH ₂ -0- P-0-CH ₂ - CH ₂ -N ⁺ (CH ₃ ) 3 O

)

where R-ι and R ₂ may be the same or different from each other, they are C12-C22 hydrocarbon chains, comprising between 0 and 8 cis double bonds, preferably between 0 and 6 cis double bonds.

In a more preferred embodiment, R-ι and R ₂ are selected from the list comprising oleic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, arachidonic acid eicosapentaenoic acid and docosahexaenoic acid, preferably oleic acid, linoleic acid, acid linolenic, palmitic acid, stearic acid.

Among the phospholipids comprising these fatty acids are soy lecithin, egg lecithin, lecithin from other seeds such as pumpkin, etc. and lecithin with omega 3 phospholipids of different origin, krill phospholipids, crustacean phospholipids, seal, etc.

In a particular embodiment of the present invention, the phospholipid has the following fatty acid composition: Fatty acid in R1 and / or R2% in oleic acid phospholipid 6-13

linoleic acid 61-71

Linolenic acid 4-7

10-15 palmitic acid

stearic acid 1, 5-3.5

arachidonic acid 0-2

In another particular embodiment of the present invention, the phospholipid has the following fatty acid composition:

This last particular embodiment is especially useful when liposomes of very high diffusion and exchange capacity, permeability and flexibility are desired. Preferably the phospholipid is selected from the group comprising soy lecithin, egg lecithin, pumpkin seed lecithin, krill lecithin and any of its mixtures, more preferably soy lecithin, although it can also be from egg or other source with high content in phosphatidylcholine. Preferably the minimum phosphatidylcholine content of the lecithins or phospholipids necessary for the manufacture or production of the liposomes of the invention is between 75% and 99.5% of the total lipid, more preferably between 96.5 and 98.5% of total lipid. In addition to phosphatidylcholine, the phospholipid bilayer of the liposome can be composed, in a proportion less than or equal to 25%, of phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, or any of its mixtures and / or degradation products such as lysophosphatidylcholine. In another preferred embodiment, the bile salt is selected from the group comprising sodium colalate, sodium deoxycholate, sodium glycocholate, sodium taurocholate, sodium taurodeoxycholate, sodium ursocholate and sodium chenoxycholate, more preferably sodium cholate.

In another preferred embodiment, the molar ratio of phospholipids / bile salt is between 1, 5 and 10, preferably between 2.5 and 5, and more preferably between 3 and 4. In another preferred embodiment, the composition further comprises an alcohol selected from the list comprising ethanol, propanol, isopropanol and any of its mixtures, preferably 96% pharmaceutical grade ethanol.

In another embodiment of the first aspect of the present invention, the ratio of alcohol / phospholipid or lecithin in volume / weight is between 0.25 and 3, preferably between 0.75 and 2.5, and more preferably between 1 and 2.

In another preferred embodiment, the aqueous phase comprises pure water or water with a salt selected from the list comprising sodium chloride, potassium chloride, magnesium chloride, sodium iodide and any mixture thereof, preferably sodium chloride. Preferably, the salt concentration is less than 9 grams per liter, more preferably between 0 and 0.9 grams per liter. However, its production with concentrations higher than 0.9% NaCl, or including different salts, especially salts in the concentration of seawater, with 35 to 38 g of salts per liter of water would also be possible. Preferably the aqueous phase comprises pure pharmaceutical quality water.

In another preferred embodiment, the composition further comprises glycated serum, or other sugars such as mannitol.

The liposome composition may also contain or comprise excipients. The term "excipient" refers to a substance that aids in the administration (absorption) of any of the components of the product of the invention, stabilizes said components or aids in the preparation of the pharmaceutical composition in the sense of giving it consistency and thus stabilizing the suspension, or provide odors that make it more pleasant. Thus, excipients may have the function of maintaining joined components such as starches, sugars or cellulose, smell or deodorize function, color function, drug protection function. Therefore, the term "excipient" is defined as that matter that, included in the galenic forms, is added to the active ingredients or to their associations to help their administration, penetration into the skin or pimples, enable their preparation or formulation and stabilize it , modify its organoleptic properties or determine the physicochemical properties of the pharmaceutical composition and its bioavailability. The "pharmaceutically acceptable" excipient must allow the activity of the compounds of the pharmaceutical composition, that is, to be compatible with said components. Examples of excipients are binders, fillers, disintegrators, lubricants, flavorings or flavors and dyes. More specific non-limiting examples of acceptable excipients are starches, sugars, hyaluronic acid, xanthan, glycerol, xylitol, sorbitol, or glycerin among others. In addition, the composition of the invention may also comprise various antioxidants, such as vitamin E, vitamin A and vitamin C, which would primarily provide a chemical stabilizing effect of the composition, and may also contain perfumes or deodorizing substances. The composition may also contain one or more additives such as buffers, surfactants, thickening agents, preservatives, or any of their mixtures.

The composition may also contain one or more synergistic or complementary actives of the element sulfur action such as, for example, antibacterials, antiseptics, hair growths, astringent, keratolytic, exfoliants, oxidoreductive assets, etc.

Another aspect of the present invention relates to the use of the composition of the invention for the manufacture of a medicament or a cosmetic or a dietary or a nutraceutical.

Another aspect of the invention relates to the use of the composition of the invention for the manufacture of a medicament for the topical treatment and / or prevention of a disease that is selected from seborrheic dermatitis, acne, rosacea, pityriasis versicolor, dermatophytosis. superficial, scabiosis, perioral dermatitis, warts, folliculitis, psoriasis, eczema, pruritus, pigmentation problems or spots on the skin, and keratosis.

Another aspect of the invention relates to a method of treatment and / or prevention of a disease that is selected from seborrheic dermatitis, acne, rosacea, pityriasis versicolor, superficial dermatophytosis, scabiosis, perioral dermatitis, warts, folliculitis, psoriasis, eczema, pruritus, pigmentation problems or spots on the skin, or keratosis, which comprises the administration of the composition of the invention, preferably topically, to a patient in need.

Another aspect of the invention relates to a process for obtaining the composition of the invention comprising the following steps:

a) preparing an aqueous solution comprising the bile salt,

b) preparing an alcohol solution comprising the phospholipid,

c) dilute by simple agitation the sulfur flower, sulfur element, in said lipid-alcohol mixture of step (b),

d) jointly homogenize the solutions obtained in steps (a) and (c); Said homogenization can be carried out by different mechanical means, stirring, shearing, extrusion, etc. or a mixture of them.

e) optionally performing a filtering stage of the product obtained in step (d), preferably sterilized, that is, filtering with simultaneous sterilization, f) diluting the liposomal concentrate with sulfur element, obtained following steps a) through e), in water without or with preservative, such as 0.5% phenoxyethanol, until the preparation or formulation has the desired concentration of sulfur element in liposomal form.

g) optionally the product can be used in concentrated form.

The product in its concentrated form obtained in steps (d) or (g) can be packaged and stored for later use mixed with other components.

Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention. EXAMPLES

The invention will now be illustrated by tests carried out by the inventors, which shows the use of the compositions of the invention as an active principle in the treatment of different dermatological disorders, both human and veterinary.

The following examples and figures are provided by way of illustration, and are not intended to be limiting of the present invention.

Example 1.

Obtaining a sulfur liposome composition element. 80 g of soy lecithin in 80 ml of 96% ethanol were dissolved by simple stirring. 320 mg of Sulfur Flor was dissolved by simple stirring in the above mixture, soy lecithin / ethanol. 1.1 g, 20 g of sodium cholate in 640 ml of water were dissolved by simple stirring. The solutions obtained were mixed and homegenized with a homogenizer and subsequently subjected to sterilization or filtering by 0.45 and / or 0.2 microns with simultaneous sterilization, obtaining a very fine and homogeneous dispersion and suspension of liposomal particles. These particles may or may not be diluted to the desired concentration of administration, for example in a 1/10 ratio to a sulfur concentration of 0.04 mg / ml. The preparation can be sterilized by 0.45 or 0.2 microns also in its diluted form. This possibility allows you to manage the products without adding preservatives. To this final preparation can be added or not preservative, odorizing substances ... This preparation is preferably carried out under vacuum or inert atmosphere to avoid degradation of the different components. Example 2

Dermatological activity tests performed with topical liposome sulfur.

During a period of six months, functional activity tests of the research product, the invention, were carried out cutaneously in volunteers with seborrheic dermatitis, seborrhea and acne. The final product used as a research product had the following composition, presentation and dosage. Composition of the final product:

- Soy Lecithin 10.00 mg / ml

- Ethanol 7.89 mg / ml

- Sodium Colato 1, 4 mg / ml

- Sulfur Flor 0,04 mg / ml

- Water, sufficient quantity up to 1 mi

- pH of the suspension 7.25 Presentation:

Aqueous suspension of sulfur sterilized liposome element at a concentration of 0.04 mg / ml of sulfur, without preservative, packed:

1- in a 20 ml / u topaz glass vial with a graffiti atomizer head.

2- in a 125 ml volume topaz glass bottle with dropper

Posology:

The sufficient amount of product was applied with the atomizer to moisten the area to be treated and perform a light massage to distribute it and make it penetrate better. In most cases, a daily application was performed, preferably at night. In some cases, the application was repeated, spacing them in time approximately 12 hours, for example, apply in the morning and at night.

Results: a. 1) Seborrheic dermatitis:

The final research product, with the composition and dosage mentioned above, was tested in eight adult volunteers, between 48 and 76 years of age, affected by seborrheic dermatitis with an important inflammatory and scaling component, mainly in the nasogenian groove area and front. The clinical trial of the final research product was performed in treatments lasting 30 days for each patient. The treatment was performed on an outpatient basis and there were two control visits per patient, one at the beginning of the treatment, with the delivery of the product under study, and, another at the end of the treatment, with the collection of data and opinions. Within a few days, (2 or 3), of the beginning of the treatment all the patients noticed the proper functioning of the product and reported a rapid decrease in the secretion or amount of fat from their skin, accompanied by a significant decrease in the inflammatory component and by both of the redness or erythema thereof, typical of seborrheic dermatitis. In patients with a significant skin scaling component (seven of the eight) there was also a rapid reduction in the intensity and number of scales, reaching, in a relatively short period of time, 2 to 7 days, at their total disappearance in all cases. The final research product and its presentation had an important degree of acceptance by the patients. All of them reported and praised the good organoleptic characteristics of the product, as well as its ease of handling and application. The final research product had a very good skin tolerance and in no case did any adverse reaction occur. to. 2) Alopecia + Seborrheic dermatitis:

The same final research product with the same dosage but with a presentation in 125 ml volume topaz glass containers with droppers, was used in five other voluntary patients, between 38 and 57 years of age, affected by androgenic alopecia with a important seborrheic component. In these cases the product was associated with other preparations or anti-hair loss / crepe products such as the 2% topical minoxidil lotion, in all cases with significant success. The ease of application, texture and organoleptic characteristics were liked by all volunteers. The product had a very good tolerance and in no case adverse reactions occurred. b) Acne:

The final research product, sulfur liposome element, was tested or tested in twelve voluntary patients of both sexes, between the ages of 16 and 24, affected by acne of varying severity. The final research product has the same dosage and presentation described in the "presentation" section.

The clinical trial of the final research product was extended over a period of six months and 21-day treatments were performed on each patient. The treatment was performed on an outpatient basis and there were two control visits per patient, one at the beginning of the treatment, with the delivery of the product, and, another at the end of the treatment, with the collection of data and opinions. Between the second or third day of the beginning of treatment, all patients could see an improvement in their general picture. To a greater or lesser degree, all patients noticed and reported a decrease in the amount of fat and skin luster, accompanied by a decrease in the inflammatory or erythematous component of comedones.

In the following days the comedones were decreasing in size and, over time, approximately between five and nine days after the start of the treatment, the decrease in their number also began. In some cases, eight, the decrease in the number of comedones was very important until it almost disappeared in its entirety during the period of treatment.

The product was very well tolerated by all patients, and, except for some dry skin that occurred in three of the cases, there were no allergic or other adverse reactions.

All patients appreciated and appreciated the ease of use of the final product, as well as its texture and organoleptic characteristics, especially its low odor.

The final product and the liposomes in the contents, as well as in the concentrate, remained physically stable throughout the period that the tests lasted and remain stable a year and a half later.

Example 3

Microbiological study: bactericidal activity of liposome sulfur against sulfur powder. Objective:

Compare in the laboratory the bactericidal activity of liposome sulfur (S-LIP) versus that of powdered sulfur (S-POL) in order to demonstrate that the liposome form has a bactericidal activity superior to powdered sulfur.

Method: From cultures of known concentrations of bacterial strains representative of the human epidermis flora or causing acne-related or cutaneous infections (S.aureus, P.aeruginosa, coliforms such as E.coli) the bactericidal effect of the two was compared sulfur forms (S-LIP and S-POL).

Materials and media:

Working bacterial strains: S. aureus, P. aeruginosa, E.coli

5ml tubes sterile

Sterile water (H ₂ 0)

- Physiological serum (SF)

Peptone Water (PEP)

Nutritive Agar (AN)

Instruction:

1- A bank of dilutions in SF of each of the aforementioned bacterial strains was prepared so as to obtain known concentrations of each of them. The bacterial concentrations for each strain that will face both substances are: 10 ⁶ - 10 ⁵ - 10 ⁴ - 10 ³ CFU ^* / 0.1 ml.

* UFC: Cologne Training Units.

2- It started with S-LIP 0.4 g / 1 OOml.

3- The following dilution was prepared with sterilized water from S-POL: 0.4 g / 100ml

4- In sterile tubes preparations A and B were dispensed together with the bacterial strains according to the following table:

S.aureus A (S-LIP 0.4) B (S-POL 0.4)

10 ⁶ (0.1 ml. 10 ⁶ + 2ml. A) x 2 (0.1 ml. 10 ⁶ + 2ml. C) x 2

10 ⁵ (0.1 ml. 10 ⁵ + 2ml. A) x 2 (0.1 ml. 10 ⁵ + 2ml. C) x 2

10 ⁴ (0.1 ml. 10 ⁴ + 2ml. A) x 2 (0.1 ml. 10 ⁴ + 2ml. C) x 2

10 ³ (0.1 ml. 10 ³ + 2ml. A) x 2 (0.1 ml. 10 ³ + 2ml. C) x 2

E.coli A (S-LIP 0.4) B (S-POL 0.4)

10 ^B (0.1 ml. 10 ^B + 2ml. A) x 2 (0.1 ml. 10 ^B + 2ml. C) x 2 10 ^b ^(b + 2ml 0,1ml.10. A) x 2 (0,1ml.10 ^b + 2ml. C) x 2

^{April 10} ⁽⁴ + 2ml 0,1ml.10. A) x 2 ⁽⁴ + 2ml 0,1ml.10. C) x 2

^{Mar. 10} ⁽³ + 2ml 0,1ml.10. A) x 2 (0,1ml.10 ³ + 2ml. C) x 2

P.Aeruginosa A (S-LIP 0.4) B (S-POL 0.4)

^{June 10} ⁽⁶ + 2ml 0,1ml.10. A) x 2 ⁽⁶ + 2ml 0,1ml.10. C) x 2

10 ^b ^(b + 2ml 0,1ml.10. A) x 2 (0,1ml.10 ^b + 2ml. C) x 2

^{April 10} ⁽⁴ + 2ml 0,1ml.10. A) x 2 ⁽⁴ + 2ml 0,1ml.10. C) x 2

^{Mar. 10} ⁽³ + 2ml 0,1ml.10. A) x 2 (0,1ml.10 ³ + 2ml. C) x 2

Table 1

Positive control: 2ml were dispensed. of physiological serum with each bacterial strains at the different concentrations, according to the following table:

Table 2 - All tubes were mixed in vortex and incubated at 37 ° C for 48 hours. - 0.1 ml were inoculated. of each tube in an AN plate. - All plates were incubated at 37 ° C for 24 hours.

- The results obtained were recorded: (Growth - C / No growth - NC):

Table 3

Table 4

Conclusions:

According to the results obtained, liposome sulfur has a bactericidal activity superior to that of powdered sulfur for all strains that have been tested (S. aureus, P.aeruginosa and E.coli), since in the test with the liposome form no growth was obtained at inoculum concentrations of up to 10 ⁶ CFU in all cases.

In the case of the sulfur powder test, bacterial growth was obtained for the three strains tested at inoculum concentrations of 10 ⁵ CFU and 10 ⁶ CFU.

Claims

1. - A composition comprising an aqueous phase liposome suspension characterized in that it comprises:

^■ phospholipids with a phosphatidylcholine ratio between 75 and 99.5% by weight,

^■ at least one bile salt,

^■ at least one alcohol and

^■ water,

and because liposomes contain sulfur element incorporated into the phospholipid bilayer.

2. - The composition according to claim 1, wherein the phospholipids have the formula (I):

OR

Ri-C-0-CH ₂ R ₂ -C -0-CH O

O CH ₂ -0- P-0-CH ₂ -CH ₂ -N ⁺ (CH ₃ ) 3 O

) where R-ι and R ₂ can be the same or different from each other, they are C12-C22 alkyl or alkenyl groups, comprising between 0 and 8 cis double bonds, preferably between 0 and 6 cis double bonds.

3. - The composition according to claim 2, wherein R-ι and R ₂ are independently selected from the list comprising oleic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid.

4. The composition according to claim 3, wherein R-ι and R ₂ are independently selected from the list comprising oleic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid.

5. The composition according to any of the preceding claims wherein the phospholipids comprise a proportion less than or equal to 25% by weight of phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine or any of their mixtures.

6. - The composition according to any of the preceding claims, wherein the phospholipids are selected from the list comprising soy lecithin, egg lecithin, pumpkin seed lecithin, krill lecithin and any of their mixtures.

7. - The composition according to the preceding claim, wherein the phospholipid is soy lecithin.

8. - The composition according to any of the preceding claims, wherein the bile salt is selected from the list comprising sodium colalate, sodium deoxycholate, sodium glycocholate, sodium taurocholate, sodium taurodeoxycholate, sodium ursocholate and sodium chenoxycholate.

9. - The composition according to the preceding claim wherein the bile salt is sodium cholate.

10. - The composition according to any of the preceding claims, wherein the molar ratio of phospholipids / bile salt is between 1, 5 and 10.

eleven . - The composition according to the preceding claim, wherein the molar ratio of phospholipids / bile salt is between 2.5 and 5.

12. - The composition according to the preceding claim, wherein the molar ratio of phospholipids / bile salt is between 3 and 4.

13. - The composition according to any of the preceding claims, wherein the alcohol is selected from the list comprising ethanol, propanol, isopropanol and any of their mixtures.

14. - The composition according to the preceding claim, wherein the alcohol is ethanol.

15. The composition according to any of the preceding claims, wherein the proportion of alcohol / phospholipid in volume / weight is between 0.25 and 3.

16. The composition according to the preceding claim, wherein the proportion of alcohol / phospholipid in volume / weight is between 0.75 and 2.5.

17. - The composition according to any of the preceding claims, wherein the composition is presented in a form suitable for topical, oral, intravenous, intramuscular, intraperitoneal, subcutaneous, topical, nail, capillary or inhalation administration.

18. - The composition according to the preceding claim wherein the composition is presented in a form suitable for topical, nail or capillary administration.

19. - The composition according to any of the preceding claims, wherein the liposomes have an average diameter between 40 nm and 500 nm.

20. - The composition according to the preceding claim, wherein the liposomes have an average diameter between 60 nm and 180 nm.

twenty-one . - The composition according to the preceding claim, wherein the liposomes have an average diameter between 75 nm and 145 nm.

22. The composition according to any of the preceding claims wherein the concentration of phospholipids is 10-200 g per liter of composition.

23. - The composition according to the preceding claim wherein the concentration of phospholipids is 50-150 g per liter of composition.

24. - The composition according to the preceding claim wherein the concentration of phospholipids is 90-1 10 g per liter of composition.

25. - The composition according to any of the preceding claims wherein the aqueous phase is pure water, pharmaceutical or cosmetic quality.

26. The composition according to any of the preceding claims wherein the aqueous phase comprises a salt selected from the list comprising sodium chloride, potassium chloride, magnesium chloride, sodium iodide and any of their mixtures.

27.- Use of the composition according to any of claims 1 to 26 for the manufacture of a medicament.

28.- Use of the composition according to any of claims 1 to 26 for the manufacture of a medicament for the treatment and / or prevention by topical route of seborrheic dermatitis, acne, rosacea, pityriasis versicolor, superficial dermatophytosis, scabiosis, perioral dermatitis, warts, folliculitis, psoriasis, eczema, pruritus, excess pigmentation and keratosis.