WO2001008632A2

WO2001008632A2 - Colloidal composition for esthetic correction

Info

Publication number: WO2001008632A2
Application number: PCT/BR2000/000085
Authority: WO
Inventors: Mateus Sommer Neto
Original assignee: Mateus Sommer Neto
Priority date: 1999-08-02
Filing date: 2000-08-01
Publication date: 2001-02-08
Also published as: AU6142500A; JP2003519093A; BR9904471A; CA2378155A1; WO2001008632A3; EP1200057A1; MXPA02001210A

Abstract

The present invention refers to a method and a chemical composition of carboxy glyconate hydrolactic or magnesium, to be employed with the aim of correcting dermatological disorders. More specifically, the present invention refers to a composition to be applied, in humans, for the correction of dermatological imperfections, such as elimination of wrinkles, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear lobe, depressions in the gluteal region, senile spots and alterations of the skin associated to natural aging, or through the action of external agents, such as sunlight.

Description

COLLOIDAL COMPOSITION FOR ESTHETIC CORRECTION FIELD OF THE INVENTION

This invention refers to a method and a chemical composition to be employed with the aim of correcting dermatological disorders. More specifically, the present invention refers to a composition to be applied, m humans, for the correction of dermatological imperfections, such as elimination of wrinkles, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear looe, depressions in the gluteal region, senile spots and alterations of the skin associated to natural aging, or through the action of external agents, such as sunlight. BACKGROUND OF THE INVENTION Human skin, when intact, is a barrier for many natural and synthetic substances. Cosmetic and pharmaceutical agents can be pharmacologically effective through oral administration or other systematic administrations of these agents, however, many agents presently known are barely or totally ineffective for topical application to the skin.

Topical effectiveness of pharmaceutical agents depends on two major factors: (a) the biological availability of an active ingredient m the topical preparation, and, (b) the absorption, penetration and distribution of the active ingredient m the area of the skin to be treated.

The formation of wrinkles may be due to natural aging and/or damages caused by the sun. A majority of the fine crinkles on the face are caused by natural or congenital aging, whilst the deeper wrinkles on the face are the consequence of a skin with acne or action of the sun. Despite the real mechanism of the formation of wrinkles remaining unknown, the formation of the fine visible wrinkles due to the reduction in the number and diameter of the elastic fibers in the papillary derails is accepted, as well as the atrophy of the dermis and reduction of adipose tissue.

Histopathological and electronic microscopy studies indicate that the tnicker wrinkles are caused due to an excessive deposit of aonormal elastic materials in the superior dermis and thickening of the skin.

Despite the fact that imperfections of the skin do not possess any physiological significance, these imperfections may cause serious physiological problems to the human being that possesses them. In many societies, the bearers of these imperfections dislike showing the signs of aging, and wrinkles are the greatest preoccupation of the human race since time immemorial . Implants and grafts presently available used to correct depressions and corporal imperfections demonstrate the difficulties in handling, complications and deficiency n long term results.

It is known in the state of the art the use of various vehicles with the aim of correcting esthetical imperfections. One of these vehicles is bovine collagen, which is a long chain protein with a high risk of allergy and passive to transmit viruses, such as bovine encephalitis, or, also, lasting future complications, such as inflammatory processes, intense allergic phenomena and, mainly, autoimmune diseases (for instance, rheumatoid arthritis) . Another vehicle also known in the state of the art is liquid silicone, however liquid silicone produces siliconomas, and has not been entirely approved by the authorities for medical use.

Amongst the classic treatments for the elimination of wrinkles, estnetic surgery may be mentioned, which, wnilst eliminating wrinkles, presents numerous disadvantages, including the scars left on the skin by tnese surgeries, considering that these scars are, frequently, very difficult to occult . Another method for eliminating wrinkles includes tne use of implants made of various materials. For instance, the Brazilian patent application PI 9503052-2 filed on 22 June 1995, in the name of Miguel Marques Oliveira Jr., incorporated herein for reference. The PI 9503052-2 describes an alloplastic surgical implant for increase and/or esthetic modeling for esthetic definition of the forehead. A prosthesis built m accordance with the PI 9503052-2, is made in semi-rigid solid silicone, and implanted m the frontal region for the correction of deficiencies, however, the disadvantages of this implant are numerous and include rejection, allergic reactions and the effectiveness is limited to a restπcteα area. Brazilian patent PI 8907235-9, granted on the 24 September 1996 to Dr. Martin Lemperle, refers to compositions for use m esthetic medicine, and is incorporate herein for reference. The PI 8907235-9 describes a composition for filling imperfections at skin level. This composition employs polymethylmetacrilate microballoons in a suspension medium. The suspension medium consists of water, alcohols, or a mixture of these, and also a tensoactive, such as Tween 80. However, as this composition is not stable, it requires to be kept under rigorous storage conditions and under constant refrigeration.

It is now Deing discovered that dermatological imperfections may oe corrected without presenting the disadvantages related in the state of the art. SUMMARY OF THE INVENTION The present invention refers to a method and a chemical composition, particularly to be employed with the ai^ of correcting dermatological disorders, however, that may be used for any other purpose in reconstructive surgery.

One objective of the present invention is to present a method and a composition possessing a perfect combination of materials, which are practical concerning storage, not requiring refrigeration, are easily handled by the doctor, do not induce allergies or infections, are not cancerigenous, are biocompatible and, finally, are low cost. One of the main objectives of the present invention is to offer a new method and composition to correct dermatological disorders, such as the elimination of wrinkles m the skin of humans, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear lobe, depressions in the gluteal region, senile spots and alterations of the skin associated to natural aging, or through the action of external agents, such as sunlight .

The composition of the present invention is applicable through the injection of the composition immediately below the area of the skin where the dermatological imperfections occur. The composition of the present invention is not, substantially, absorbable by the human body, and, consequently, will not be rejected by the human oemg, and will also not cause allergic reactions.

In addition, the method of injecting the composition, in accordance with the present invention, is easily and dependably applicable, without requiring the anesthetization of the patient . DETAILED DESCRIPTION OF THE INVENTION

The composition, in accordance with the present invention, is composed of a colloidal formulation termed carboxy glyconate hydrolactic of magnesium. This formulation consists of:

(a) approximately 5 to 70g of an ether of cellulose;

(b) approximately 800 to 1300ml of Ringuer lactate; (c) approximately 2 to 20g of EDTA;

(d) 20 to 120ml of calcium glyconate; and,

(e) optionally, between 0 and 10% xylocaine. The quantities of the ingredients may vary within the optimal ranges until adequate viscosity is attained.

Amongst the polymers of ether of cellulose, those with low molecular weight or low viscosity are preferable, such as: hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxybutylcellulose , hydroxypropylmethy1cellulose, carboxymethylcellulose of sodium, carboxymethylcellulose of calcium, methylcellulose and/or ethylcell αlose .

The Ringuer lactate is an inert solution composed of sodium chloride, potassium chloride, dihydrated calcium chloride, sodium lactate and water.

The EDTA, termed tπhydrated ethyleneαia me tπsodium tetracetate is cnemically inert, and consequently, appropriate for medical use, oecause apart from not decomposing in other substances, it does not react with other chemical bodies whether in the composition of the invention or in the human body. Its chemical formula is Cio-Hiβ-N∑-Oβ .3Na .

The preparation process of the composition of the present invention consists m mixing all the ingredients in stainless steel tubes, followed by agitation. The temperature of the mixing may vary between 30°C and 70°C, preferably at around

50°C, and for a time period for approximately 3 hours. This procedure is repeated twice more, until obtaining the final product .

At this stage a colloidal solution is obtained.

The resulting colloidal product is then filtered, and its pH is verified so as to present a value of approximately 6.0.

The density of the product shall present a value equal to 1.

The resulting product, that is, the carboxy glyconate hydrolactic of magnesium composition, already having the pH and αensity required, __s injected nto vials of 20ml, and after labeling, are ready for distribution.

To illustrate the present invention, experiments in animals were performed. These experiments were based on the investigation of the reactions occurring m the issues of the animals after the application of the exogenous materials m accordance with the present invention. EXPERIMENT I

Comparative histological sτuαy of the cellular alterations and of the extracellular matrix resulting from the injectable implant of the carboxy glyconate hydrolactic of magnesium colloid m the hypodermis of albino rats. Preliminary results

In Experiment I the possible different tissue reactions resulting from the injection of carboxy glyconate hydrolactic of magnesium were evaluated m a group of 108 rats, obtaining samples in the 1^st, 2^nd, 4^th, 8^th and 16^th weeks, with another 20 rats injected with a physiological solution of sodium chloride, kept m identical conditions to act as a control. Materials and Method

Seventy-two adult rats, albino, females of the istar breed, were superficially anesthetized through the inhalation of ether, trichotomized on the back (interscapular regionj with an electric hair clippers, selecting, on the median dorsal line, an area where two surgical stitches were applied, placed sagitally, with black l ne (having a distance of 1.5cm between them) . On the back, m the interscapular region, a first group of 36 rats was injected with 0.5ml of the camoxy glyconate nydroiactic of magnesium colloida_L comoositior, and the rats were placed in three appropriate plastic cages 13cm each) with a bottom of sterile wood shavings and supplying them with food and filtered water ad l ibi tum . A second group of 36 rats was, treated similarly, however a physiological solution of sodium chloride was injected and these rats were kept m identical conditions to act as a control.

After 1, 2, 4 and 8 weeks, 14 animals (sever injected with carboxy glyconate hydrolactic of magnesium and seven in^jected with the physiological solution were put down in oeep anesthesia with ether, and, then had fragments of skin, kidney, liver, spleen, heart and lungs removed, at the same time that their principal lymph nodes were examined for eventual histopathological studies, in the case of growtn being found. In the 16^th week, the remaining animals were put down. The fragments were fixed in a solution of formaldehyde at 10% in PBS, at 4°C, for a period of 3 to 5 days, after which they were washed for 15 minutes m running water. In addition, a second fragment of skin from the region injected was removed from each animal, and fixed for 4 hours m a solution of paraformaldehyde at 5% n PBS for later lmmuno-histochemical study and, after washing in PBS, the fragment was treated m the same way as the others for histological processing.

All the fragments were then dehydrated m solutions of increasing concentrations of ethanol, diaphanized by xylene, to be later impregnated and, finally, included m paraffin. Cuts of 5 micrometers were obtained using a Spencer 820 rotative microtome from American Optical Co., and, those previously fixed m formaldehyde-PBS, after being mounted on histological slides, were dyed by the Hematoxillm-Eosin technique. The preparation could then be examined by an Optiphot model Nikon microscope, coupled to an automatic AFX II photometer with a 35mm camera, thus obtaining photomicrographies on 100 ASA Kodacolor film from Kodak.

During the first phase of observations, it is possible to observe in tie first week, througn the visualization of the skin from the animals injected with carboxy glyconate hydrolactic of magnesium, that droplets of colloid

(identifiable as amorphous and basophilic material) are surrounded oy mononuclear cells, which develop intense infiltration m the area of the injection around the annexes below the epidermis. In the second week, the infiltration persists and macrophages can already be observed around the droplets of colloid, as well as giant multmuclear cells. In the fourth and eighth weeks, macrophages appear, either molding incipient capsules or infiltrating between the colloi^'d droplets. Finally, in the 16^th week, macrophages, fibroblasts and conjunctive matrix surround conglomerates of colloid, still distinguishable by their basophilia. In addition, the observation of images of the retraction of the blood plasma fixed inside the blood vessels close to the endothelial surface, which could be confused with a possible presence of minute mcroballoons m the blood, become, m reality, impossible to be mistaken, as in the blood vessels of tne animals injected with NaCl, such linages are also present. No signs of neoplasia were detected m the fragments of the skin studied.

From the second week of experiment, the colloid is already retained in the granuloma, m stage of formation, present m a general manner below the muscle, but that can pass the muscular layer and reach the dermis . This granuloma evolves until constituting a sac containing the colloid inside still surrounded oy the macrophages. In tne 16" week, conjunctive matrix can be observed between the macrophages and fibroblasts as part of the conjunctive capsule surrounding the granuloma. Conclusion of Experiment I

From the observations of Experiment I, it can be concluded that the injection of carboxy glyconate hydrolactic of magnesium, in the hypodermis of albino rats, promotes the formation of a granuloma reacting to a foreign body, localized preferentially at the place of injection, that is, in the inferior part of the skeletal muscular layer (present m the rat) of the hypodermis. In some situations, however, it reaches the supra muscular region of the hypodermis, attaining deep dermis. This granuloma becomes stable around the second week, after the injection, and the carboxy glyconate hydrolactic of magnesium colloid can be visualized m an apparently stable form surrounded oy giant foreign body cells. After the eighth week, the granulomas becomes surrounded by conjunctive capsules that extend septa inwards, turning the granulomas lobulated. The presence of neoplasia was not observed. EXPERIMENT II

Comparative histological study of the cellular alterations and the extracellular matrix resulting from the injected implant of the carboxy glyconate hydrolactic of magnesium colloid in the hypodermis of albino rats. Evaluation of the matrix In Experiment II, adult female albino rats of the istar breed were used.

This Experiment II was an evaluation of tissue alterations resulting from the injected implant of carooxy glyconate hydrolactic of magnesium in 108 albino rats, injecting these 108 rats with carboxy glyconate hydrolactic of magnesium m the previously tπchotomized dorsal interscapular region. Another 20 rats were injected with a physiological solution of sodium chloride, and were kept m identical conditions to act as a control . Material and Methodology

The methodology used for obtaining the tissue of the animals is described below. The albino rats, after being put down m deep anesthesia, had fragments of skin, kidney, liver, spleen, neart and lung removed. These fragments were fixed in a solution of formaldehyde at 10% in PBS, at 4°C, for a period of 3 to 5 days, after which they were washed for 15 minutes m running water. All the fragments were then dehydrated in solutions of increasing concentrations of ethanol, diaphanized by xylene, to be later impregnated and, finally, included in paraffin. Cuts of 5 micrometers were obtained using a Spencer 820 rotative microtome from tne Amer_can Optical Co., and after oeing mounted on histological slides, were dyed by either the Hematoxillin-Eosin technique, or by the Gomori tπcnromic method. The preparation could then be examined by an Optiphot model Nikon microscope, coupled to an automatic AFX II photometer with a 35mm camera, thus obtaining photomicrographies on 100 ASA Ektachrome film from Kodak.

Samples were obtained of tissues from rats m the 1^"", 2^nα, 4^th, 8^" and 16^tr weeks, to evaluate possible differences in tissue reactions resulting from the injection of carboxy glyconate hydrolactic of magnesium.

It was verified that the injection of carboxy glyconate hydrolactic of magnesium m albino rats promotes the formation of a granuloma in reaction to foreign bodies, localized preferably in the area of the injection, that is, m the inferior skeletal muscular layer (present in the rat) of the hypodermis In some situations, however, it reaches the supra muscular region of the hypodermis, attaining deep dermis. This granuloma becomes stable around the second week, after the injection, and the carboxy glyconate hydrolactic magnesium colloid can be visualized in an apparently stable form surrounded by giant foreign body cells. After the eighth week, the granulomas becomes surrounded by conjunctive capsules that extend septa inwards, turning the granulomas lobed. The presence of neoplasia was not observed.

Through photomicrographies, it was possible to observe that when employing the composition of the present invention, collections of droplets of carboxy glyconate hydrolactic magnesium colloid, of irregular shape and showing basophilic content, were observed, either grouped between macrophages or surrounded by giant foreign body cells.

The injection of carboxy glyconate hydrolactic of magnesium colloid produced inflammatory reactions at the end of the first week filling the area with macrophages infiltrating between the droplets of amorphous and basophilic colloid, promoting a matrix deposit well visible below the muscular layer, with an abundance of reticular fibers, revealed by the green refπngency, comparable to that of the peri and endomysium.

In the second week, macrophages already form a thick layer around the collections of colloid droplets and infiltrate between colloid conglomerates, forming septa that show more extensive deposits of extracellular matrix rich in reticular fibers in the more slender ones and collagen fibers in the thicker ones. In the fourth and eighth weeks, it is possible to observe, with clarity, that when of carboxy glyconate hydrolactic of magnesium is injected, amorphous spaces occur containing basophilic material (colloid) forming separate aggregates. In the fourth week and the eighth week, the predominance of fibers with green birefringency in the interstices of the aggregates of the colloid droplets is observed, compatible with the predominance of collagen III, particular to reticular fibers.

In the sixteenth week, it is observed that the colloid granulomas show a predominance of green birefringency in the interstice with slenderer septa and yellow or red birefringency in the thicker septa and capsules. This is in accordance with the greater quantity of collagen III present in the interstice of the smaller septa. Conclusion of Experiment II

From the observations of Experiment II, it is possible to conclude that the injection of carboxy glyconate hydrolactic of magnesium, in the hypodermis of albino rats, promotes the formation of granulomas reactional to foreign bodies having the same characteristics as extracellular matrix. The granulomas thus constituted become stable around the second week after the injection and, the colloid can be visualized m an apparently stable form surrounded by giant foreign body cells. Already, in the first week, the actual macrophages when infiltrating the colloid begin to promote the septation of the future granuloma, with an initial production of extracellular matrix. The granuloma, in the second week, exhibits more advanced septation and, m the 8^th week, possesses a thick conjunctive capsule r ch in collagen I that sends septa inwards, making the mentioned granuloma distinctly lobulate. However, inside the lobules, the richness of collagen III is notable. This richness of collagen III inside the stabilized granuloma tends to indicate that the slender conjunctive fibers present there do not characterize a state of fibrosis, when thick conjunctive fibers (collagen fibers) rich in collagen I would predominate. METHOD FOR TREATMENT OF DERMATOLOGICAL DISORDERS A method to treat dermatological disorders oy filling and correcting unaesthetical alterations with the composition of carboxy glyconate hydrolactic of magnesium, in accordance with the present invention, is described below.

A composition is obtained by the process already described herein, consisting of mixing all the ingredients m stainless steel tubes, followed by agitation. The temperature of the mixing may vary between 30°C and 70°C, preferably at around 50°C, and for a time period for approximately 3 hours.

This procedure is repeated twice more, until obtaining the final product.

At this stage a colloidal solution is obtained. The resulting colloidal product is then filtered, and its pH is verified so as to present a value of approximately 6.0 and a density equal to 1. The resulting product, that is, the compositon of carboxy glyconate hydrolactic of magnesium, already having the pH and density required, is injected into vials of 20ml. For the injection of the product, it is initially necessary to identify the areas to be treated and, if necessary, to mark these areas in advance w th a patient standing upright. Next, the patient is placed in dorsal decubitus, m a seated position, at an angle of between 45 and 60 degrees, when the area to be treated is the ventral part of the head, or without ventral decubitus, when the area to be treated is the dorsal area.

The asepsis and antisepsis is then performed with the usual antiseptics.

As routine, a blocking or even local infiltration with anesthetics is performed for greater comfort and less stress for the patient during the procedure.

There is no necessity of prior tests as the product is inert and biocompatible, that is, exempt of any allergic or toxic reaction. The formulation of the composition, in accordance with the present invention, impedes contamination by fungi or bacteria.

The content of the vial is aspirated with a needle or cannula, this latter with a rhombic point. The size of the needle or cannula varies in accordance with the quantity necessary for filling the area to be treated.

The application of the composition of carboxy glyconate hydrolactic of magnesium is performed with a needle or a cannula, depending on the area to be treated. The injection of the composition should¹- avoirT The" formation of sequential bubbles, like a pearl necklace, because in this manner the result would be irregular.

Being a permanent implant, hypocorrections were performed and, when necessary, more than one or two complementary sessions can occur, with an interval of 25-30 days between each session. To obtain a better result in total safety, this time interval was established for a further injection. This interval is fundamental for the deposit of collagen and fibrin fibers, providing the necessary support for the microimplant.

Claims

1. Colloidal composition for esthetic corrections characterized by being composed of a colloidal formulation termed carboxy glyconate hydrolactic of magnesium, consisting of:

(a) approximately 5 to 70g of an ether of cellulose;

(b) approximately 800 to 1300ml of Ringuer lactate;

(c) approximately 2 to 20g EDTA;

(d) 20 to 120ml of calcium glyconate; and, ιe) optionally, between 0 and 10% xylocame .

2. Colloidal composition m accordance with Claim 1, characterized by the fact that the ether of cellulose is a polymers of ether of cellulose selected from hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxybutyl cellulose, hydroxypropylmethyl cellulose, carboxymethyl cellulose of sodium, carboxymethyl cellulose of calcium, methylcellulose and/or ethylcellulose .

3. Colloidal composition m accordance with Claim 1 characterized by the Ringuer lactate being an inert solution composed of sodium chloride, potassium chloride, dihydrated calcium chloride, sodium lactate and water.

4. Composition in accordance with Claim 1 characterized by being applied, in humans, for the correction of dermatological imperfections, such as elimination of wrinkles, correction of the nasogenian furrow, labial reduction and contour, keratosis, augmentation of the malar region, reduction of the ear lobe, depressions m the gluteal region, senile spots and alterations of the skin associated to natural aging, or through the action of external agents, such as sunlight.

5. Process for the preparation of the colloidal composition for esthetical correction characterized by including the stages of:

(a) mix an ether of cellulose, a Ringuer lactate solution, EDTA and calcium glyconate;

(b) agitate the ingredients of stage (a) , at a temperature varying between 30°C and 70°C, for a time period for approximately 3 hours;

(c) repeat the procedures of stage (b) twice;

(d) filter the product obtained in stage (c, , and

(e) adjust the pH of the filtered product to an approximate value of 6.0.

6. Process in accordance with Claim 5 characterized by also including the addition of xylocaine, as an optional ingredient .