FR2537585A1 - Method and product for stimulating the synthesis of the melanin pigment of the skin, especially for the treatment of vitiligo, and method of preparing this product - Google Patents

Abstract

A compound of the alpha-lipoprotein type for stimulating the synthesis of the melanin pigment of the skin is prepared in order to cure vitiligo without toxic effect, with an efficacy which is irreversible after ceasing the treatment, by extracting this compound from human placental cotyledons by means of organic solvents, followed by filtration and then freeze-drying and dissolving in ethyl alchol. The compound is topically applied to the skin and then the treated part is exposed once daily to sunlight or ultraviolet light for 15 min.

Description

"Method and product for stimulating the synthesis of the melanin pigment of the

  skin, in particular for the treatment of vitiligo, and process for the preparation of this product "

  The present invention relates to a method and a pro-

  duit to stimulate the synthesis of the melanin pigment of the

  skin, useful mainly for the treatment of vitili-

  go, dermatological disease produced by the absence of pig-

  melanin in various regions of the human body.

  Various chemical compounds are known, of vegetable origin.

  tale or semi-synthetic, known as psoralens which, when ingested by the patient or applied

  on the skin of it, concentrate in the melano-

  phores, i.e. the cells that produce the pig-

  dermal by absorbing the energy of ultra-

  purple and producing the dermal (melanic) pigment.

  Unfortunately these known chemicals present

  have the disadvantage of being toxic; when they are applied to the skin, they can produce dermatosis and necrosis of the skin; if such compounds are ingested,

  they can produce chemical disorders.

  Such compounds are described in the following publications: Arnold, M J Jr "Psoralens and Suntan" Hawaii Med J 1957

16-391.

  Becker, S W Jr "Methods of increasing skin pigmentations".

J Sec Cosmetic Ehem 1958-9-80.

  Fitzpatric, T B "Pigmentary Diseases" Current Therapy

1958, p 514.

  Becker, S W Jr "Effects of 8-Methoxy Posralen and ultra-

  violet light in human skin "Science 1958-127-878.

  Sidi, E Planat, P "Practical considerations on actual

  treatment of vitiligo "Revue of Medicine, 1968-23 Dec.

  To date psoralens are the best

  known drugs for the treatment of vitiligo or leu-

  codermia, but they have, in addition to their toxicity, a certain number of disadvantages, in particular: a very high cost, a slow and reversible effect when the treatment is stopped, some difficulties of application, given that it

  is necessary to gradually increase the duration of

  position to ultraviolet radiation, and severe reactions

  Toxic tions, such as burns, can occur when applied to the skin; finally their ingestion

  risk of producing liver failure and

digestive and nervous.

  On the other hand, the product according to the invention stimulates the

  synthesis of the melanin pigment of the skin and the reproduction

  tion of the cells producing the melanic pigment, as is

  proven by phaxracological tests, without the aforementioned drawbacks.

  Toxicological, teratological and clinical trials

  that have also proven that there are no reactions

  severe condaries with the product according to the invention.

  The product according to the invention is easy to obtain and easy to use, since it only requires a short time

  exposure of the sick person to ultra-

  purple; its effectiveness begins very quickly (after 15 to 20 days) after the initial treatment, the color of the skin resulting from the treatment with the product according to the invention is identical or substantially identical to the color of the normal skin of the sick person (before illness) and the intensity of healthy skin color does not increase, as is the case when using

psoralens.

  Since the product according to the invention is obtained

  held from human tissue, it does not cause

  immunological reaction and has no contraindications

  cation relating to age, sex or race.

  The product according to the invention is intended primarily-

  to cure vitiligo This disease is one of the most

  ancient human diseases It affects approximately

  1% of the population of the world without discrimination of age, sex or race Its etiology is unknown and

  there is no known definite drug to combat it.

  This disease is characterized by discoloration pro-

  aggressive skin of affected people, who are

  throw to stress or significant nervous tension She

  adversely affects the psychology of people with

  tints, as well as their social behavior, because 1o its symptoms appear in the form of white skin areas surrounded by a halo of hyperpigmentation of the

  -figure, trunk and other regions.

  The product according to the invention is obtained by a new

  calf process for extracting a compound from the cotyledons

  centenary humans We treat cotyledons using

  organic solvents (ethyl alcohol and benzol acid-

  ques) Then it is filtered using a Sephadex type gel.

  The product obtained contains a lypo-protein and the electrophoresis migration test makes it possible to identify it as an alpha lypo-protein. The product is lyophilized and

  -dissolved in ethyl alcohol.

  We will now give some examples of preparations

tion of the compound according to the invention.

  We start from a kilogram of human placental cotyledons after their freezing for 7 to 10 days, then they are triturated and soaked in 95% ethyl alcohol in equal parts (weight / volume). The product obtained is then filtered to through fine gauze and we leave it

decant for 24 hours.

  The pigment activity of the pro-

  said active supernatant, which was positive for 20 to 50

  days after its use on the skin of guinea pig udders.

  Then we took 20 ml in this extract

  with the pigmentation factor and 10 ml of al-

  cool ethyl saturated with 95% benzoic acid in use

a hypodermic needle.

  Saturated solution of benzoic acid / ethanol A mixture of 2 to 8 g of benzoic acid in 5 to 12 ml of 95% ethyl alcohol is prepared and this is stirred.

  mixing for 10 to 15 minutes, then it is filtered on a

  Buchner's barrel with Whatman No. 2 to 5 filter paper, then the precipitate is washed using 200 to 500 ml of water

saturated with benzoic acid.

  Saturated solution of water / benzoic acid A mixture of 2 to 10 g of benzolque acid in 300 to 1000 ml of distilled water is prepared. Then the precipitate is removed on the filter paper using a spatula.

  and then add 100 to 500 ml of pure acetone or

  tone at 30% Then the resulting product is centrifuged at 1500 revolutions / minute for 10 to 30 minutes, putting aside the

supernatant portion.

  The precipitate is washed again 2 to 10 times in u-

  using 100 to 500 ml of acetone, then it is dried under

  vacuum and at room temperature Then dissolve the pro-

  duit in 95% alcohol and we use a guinea pig or guinea pig to determine the pigmentary activity Then we filter ml of this product with biological activity by means of a Sephadex g-30 or 100 filter balanced by means of '' a buffer of sodium phosphate 5 by 10 to 5 by 10 6 M at p H 4 to 8, then a column of 1 to 3 cm in diameter and 18 to 40 cm in height is filled with it in fractions of 5 to 10 ml c / u We obtained two protein peaks, the first between 100 and

  300 mcg / ml and the second at 20-150 mcg / ml The pigmen-

  shut up was positive for the second peak She appeared

  10 to 20 days after the lyophilized mixture has been applied

  stuck on the skin of the guinea pig.

  The efficacy of the product was determined according to the invention.

  tion by clinical experiments carried out on 200

  sick people, men and women of the derma- service

  tology of Calixto Garcia Hospital in Havana The ma-

  lades presented the characteristic lesions of the vitili-

  go, scattered or localized on their body and having

  of various dimensions The age of the patients was understood

  be two and sixty four years old and they were only found

  under no treatment at the start of the experiments.

  results obtained for four years and two months by implementing the invention can be summarized as follows: total of sick people 200 (100%) total of repigmented people 168 (84%) total of completely people 63 (31.5%) repigmented total of partially 105 (52.5%) people repigmented total of non-repigmen 5 (2.5%) total of people who have 27 (13.5%) discontinued treatment

  There were no local or systemic secondary reactors

  no evidence of relapse after treatment Biopsies were performed on the basement membrane, which proved that the product according to the invention produced the synthesis of the melanin pigment in the skin by eliminating the defect which is characteristic of this disease and which is produced

  in the absence of melanin pigment in the basement membrane.

  The biopsies mentioned were carried out at the

  pathological anatomy at Calixto Garcia Hospital.

  The method of using the product according to the invention

  to cure vitiligo is as follows -

  The product according to the invention is applied, which is

  smells like a lotion, topically three times a day, rubbing the areas strongly with your fingers

  affected, until reddening Then we expose to the lu-

  sun or ultraviolet light the areas thus treated, for 15 min once a day after one

  daily applications of the product.

  The daily amount to use depends on the size

  the number of affected areas of the patient's body.

  This quantity is not critical since there is no side or toxic effect, as has been observed during the treatments carried out at the Hospital.

Calixto Garcia.

  Preclinical trials in mice, rats, guinea pigs and rabbits, with application

  topical of the product three times a day, followed by an expo-

  15 min daily exposure to sunlight, did not reveal any macroscopic or microscopic alteration, although the duration of the experiment lasted a very long time (a whole year with adult animals, 75 days with pregnant females and 15 days with newborn animals)

  As it goes without saying and as it results from elsewhere

  In addition to the above, the invention is in no way limited to those of its modes of application and embodiments which have

  been more specifically contemplated; she embraces it,

milk, all variants.

Claims (4)

  1 Process for the preparation of an alpha-type compound   hypo-protein to stimulate the synthesis of the pigment mela-   nique of the skin in order to cure vitiligo without toxic effect, with non-reversible efficacy after cessation   tion of the treatment, characterized in that this com-   posed of human placental cotyledons by means of sol-   organic fibers, then filtration followed by lyophilisa-   tion and a solution in ethyl alcohol.   2 Method according to claim 1, characterized in   that said organic solvent consists of al-   cool ethyl and / or benzoic acid.   3 Method according to claim 1 or 2, characterized   in that the filtration takes place by means of a Sepha gel   dex. 4 Compound of the alpha-lypo-protein type to stimulate the synthesis of the melanin pigment in the skin in order to cure vitiligo without toxic effect, with non-reversible efficacy after cessation of treatment, characterized in that it is prepared by the process according to any one of claims 1 to 3 '