DE2817133A1 - AGENT FOR TREATING HYPERPIGMENTAL DERMATOSES, NEW COMPOUNDS CONTAINING THEREOF AND METHOD FOR THE PRODUCTION THEREOF - Google Patents

Description

Patent attorneys

PROF. DR. DR. J. REITSTÖTTER DR.-ING. WOLFRAM BUNTE DR. WERNER KINZEBACH

BAUERSTRASSE 22. D-BOOO MUNICH AO FERNRUF (O39I 37 65 83 TELEX S21S2O8 ISAR D POSTAL ADDRESS: POST BOX 7BO.D-BOOO MUNICH 43

Munich, April 19, 1978 M / 19 081

Dr. Marcella Nazzaro Porro Via Viminale 38
Rome, Italy

Preparations for the treatment of hyperpigmentary dermatoses, therein | contained new compounds and processes for their preparation '

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The invention relates to that characterized by the claims Object. j

Dermatoses, for example skin hyperpigmentation, which often |

disfiguring, as in the case of chloasma on the face, '

not only pose an aesthetic problem, but also ι

a therapeutic problem. A basic solution ι

this problem has not yet been identified. So far _;

have only hydroquinone and its derivatives in vivo at < the treatment of skin hyperpigmentation a certain amount

Effectiveness shown. However, they bring as a side effect; the development of long-lasting hypopigmented zones

with them that can sometimes no longer be eliminated. ;

The field of hyperpigmentary dermatoses includes, however
with much more serious aspects and consequences, j certain pathological conditions that can be classified under the generic j designation of precancerous and cancerous dermatoses (for example premelanoma conditions and melanomas).
There is currently no effective therapy for these dermatoses.

The invention is based on the object of providing medicaments which can be used for the above purposes. she
in particular, are intended to normalize the color of skin suffering from hyperpigmentation and the appearance of certain shapes
Prevent and prevent cancerous and cancerous dermatoses
cause their regression.

The invention thus provides a medicament for treatment
hyperpigmentary dermatosis, which has a dicarboxylic acid
7 to 13 carbon atoms or a derivative thereof which
contains at least one reducing group in the molecule,
includes.

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According to the invention it is possible to treat hyperpigmentary dermatoses using a therapeutically effective amount administered a drug containing a dicarboxylic acid having 7 to 13 carbon atoms, a derivative thereof, which is at least has a reducing functional group in the molecule, or a pharmaceutically acceptable salt thereof, includes.

The dicarboxylic acids used in the present invention are particular selected from the following group: pimelic acid, suberic acid, azelaic acid, sebacic acid, 1,9-nonanedicarboxylic acid, 1,10-decanedicarboxylic acid and 1,11-undecanedicarboxylic acid. The derivatives can be mono- or disubstituted by a reducing functional group. To the Pharmaceutically acceptable salts include the sodium salts.

Contains in a preferred embodiment of the invention the drug is a mercapto derivative of dicarboxylic acid.

In a further preferred embodiment it is the dicarboxylic acid is azelaic acid.

The medicaments according to the invention can be administered orally or subcutaneously Injection or topical application, for example in the form of creams, ointments, pastes or lotions will.

It was found that the medicament according to the invention is particularly capable of inhibiting the formation of the skin pigment (melanin) by blocking the dopatyrosinase reaction through a reaction which proceeds according to a kind of competitive mechanism. The exact mechanism of this effect has not yet been clarified. According to the application, however, a mechanism was found which corresponds to or is related to the mechanism exerted by the active component of the medicament.

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While these observations are brought to a better understanding of the invention, they should not be taken as definitive Explanation of the mechanism at work is to be understood, nor is it intended to in any way exceed the scope of the present Limit the invention.

When investigating the behavior of a fungus pityrosporum orbiculare, which is to be understood as the cause of dermatoses that are achromatic or whitish Observations in vitro and in vivo have reasonable indications supplied to ensure that the fungus is usually present on the skin and for survival and diffusion i Metabolized fatty acids required and leads to the formation of I dicarboxylic acids, which then have an inhibiting effect exhibit on melanogenesis.

In this context, an examination of cultures of Pityrosporum orbiculare to which technical oleic acid was added as a lipid supplement found after saponification that pimelic acid, azelaic acid and 1,9-nonanedicarboxylic acid (C ₁₁ ) were also present. These were identified by gas chromatography and mass spectrometry. The Cg and C ₁₁ representatives of the dicarboxylic acids have shown considerable anti-tyrosinase activity in vitro, which was also found in dicarboxylic acids with 8, 10 and 12 carbon atoms.

The invention also relates to new derivatives of dicarboxylic acids having 7 to 13 carbon atoms, which are characterized are that they have at least one reducing group, and their pharmaceutically acceptable salts.

The derivatives have an inhibiting effect on the dopatyrosinase reaction, that of the effect of the dicarboxylic acids with 7 to 13 carbon atoms from which they are derived,

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is superior.

In a preferred embodiment, the reducing group of the dicarboxylic acid derivative is a Mercapto group.

In a further preferred embodiment, the dicarboxylic acid derivatives contain 2 reducing groups, which are preferably both times are mercapto groups.

The invention also relates to a process for the preparation of mercapto-substituted derivatives of dicarboxylic acids.

The process is characterized in that the dicarboxylic acid is brominated to add as many bromine atoms to the acid introduce how to be replaced by mercapto groups, the bromo derivative with thiourea in a solvent reacted to form the corresponding thiouronium salt and then saponified the salt thus formed to give the corresponding Obtain mercapto derivatives.

The bromination of the dicarboxylic acid can be achieved by first reacting the dicarboxylic acid with phosphorus pentachloride until the mixture has completely melted and then adding anhydrous bromine ^{to the melt with stirring at a temperature which is kept at 60 to 70 ° C.} The bromine derivative formed in this way can then be isolated in the usual way.

In a preferred embodiment for the preparation of the monomer capo derivatives of the dicarboxylic acids, 1 mol is used Dicarboxylic acid with 50 g of phosphorus pentachloride. The reaction product is then reacted with 60 ml of anhydrous bromine, the anhydrous bromine being added in small portions over about 6 hours. Here is the monobromo derivative of the dicarboxylic acid. Then add 1 mole of the

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Monobromo derivative with 50 ml of ethyl alcohol and 1.1 mol of thiourea in order to produce the corresponding alkylthiouronium salt which is then saponified to obtain the corresponding monomer capo derivative.

In a preferred embodiment for the preparation of the dimercapto-substituted dicarboxylic acid derivatives ^!

1 mol of acid with 50 g of phosphorus pentachloride and sets 120 ml of anhydrous bromine to the resulting melt in the course of 18 hours in small increments while keeping the temperature

at 90 to 100 ⁰ C holds. Then 1 mole of the dibromide obtained is reacted with 2.2 moles of thiourea in 100 ml of ethyl alcohol to form the corresponding alkylthiouronium salt, which is then saponified to give the corresponding dimercapto derivative

to obtain.

The above-described dicarboxylic acids or their derivatives or pharmaceutically acceptable salts can be in various pharmaceutically acceptable carriers, the choice of which depends on the type of treatment. So they can for example in the form of an injectable suspension of the active material in a saline solution. But if they are intended for topical administration, you can they are applied as creams. They can also be presented in an orally administrable form. The amount of active ingredient present in these drugs may vary somewhat. It is only essential that the Dicarboxylic acids or their derivatives are present in therapeutically effective amounts.

The amount of dicarboxylic acid or its derivatives or pharmaceutical compatible salts of these compounds, which are present in the agent according to the invention, varies, depending on the dosage form and / or the condition being treated. When administered orally, the amount of active ingredient can reduce the Total amount or essentially the total amount of the dose

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make shape. However, when the active substance is contained in a dosage form suitable for subcutaneous injection is, it can be about 20 to 30 wt .-%, and preferably about 25 wt .-%, based on the total weight of the Means, make it up. In the case in which the agent is present as a cream or lotion for topical administration, the Dicarboxylic acid or its derivative between about 10 and 20% by weight, preferably about 15% by weight, based on the total weight of the agent.

It has also been found that it is advantageous to include a keratolytic agent in the medicaments of the invention bring in. This is particularly the case when the agents are to be applied to the skin. As examples for keratolytic agents used in the medicament according to the invention salicylic acid, vitamin A acid, resorcinol, phenol, cresol and the like can be mentioned. The amount of keratolytic agent that is beneficially used also varies, usually making it approximately 2 to about 4% by weight, based on the total weight of the agent.

The following examples serve to explain the preparation of the derivatives according to the invention without restricting the subject matter of the invention.

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example

(a) Preparation of the alpha monobromo derivative:;

1 mole of dicarboxylic acid (for example azelaic acid) is placed in a 3 liter flask placed on a magnetic stirrer with a hot plate, and 500 g of phosphorus pentachloride are added. After the reaction has taken place (complete melting), 60 ml of anhydrous bromine are added in small portions over a period of about 6 hours with continuous stirring. The reaction temperature is kept at 60 to 70 ^° C.

When the reaction is complete, the mixture is cooled and then about 500 ml of distilled water are carefully added, heated then about 30 minutes on the magnetic stirrer and cools down.

The lower organic phase, which consists of a dark yellow oily liquid, represents the crude Monobromo derivative that is distilled in vacuo.

(b) Preparation of the mercapto derivative:

Adding 1 mol of the alpha-monobromderivats under a hood in a 2-liter flask equipped with condenser, and then at 95 ⁰ C 50 ml of ethyl alcohol and 1.1 mol of thiourea to. The mixture is heated at the boiling point for 6 hours, after which the alkyl thiouronium salt is separated off by cooling.

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In order to saponify this salt, 500 ml of 5N NaOH are added to the mixture and the mixture is kept at the boiling point under reflux for a further 2 hours. After cooling, the reaction mixture is acidified with 5N HCl and, after stirring for about 10 minutes, an oily layer forms which is removed. Extract the aqueous layer three times with ethyl ether, and outputs this ether extract to the previously removed oily layer which then over anhydrous ₀ SO. is dried. After getting the ether

Z 4

removed by distillation, the crude mercapto derivative is purified in a silica gel column.

To prepare the dimercapto derivatives, repeat the procedure of the previous example with the following

Differentiated:

1. The dibromo derivative of the dicarboxylic acid is produced like the monobromo derivative, but the amount of bromine added is doubled and the reaction times are tripled; the reaction temperature is maintained at 90 to 100 ⁰ C.

2. In the production of the mercapto derivative, double the amounts of alcohol, thiourea and Sodium carbonate used per mole of dibromide.

To the effectiveness of the agents according to the invention and their corresponding To evaluate active ingredients, both pharmacological studies and tests in vivo were carried out.

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From the combined lipid extracts (from cells and from the filtrate) of Pityrosporum orbicular cultures (strain 4709) which were grown for 20 to 30 days from a conventional synthetic medium to which oleic acid was added, obtained a saponifiable portion with considerable tyrosinase. inhibiting activity. During an investigation of this portion by means of thin layer chromatography (TLC), a fraction with Rf = 1.3 was isolated, which the Can inhibit DopH tyrosinase reaction.

Gaschromatographxsche analysis and subsequent mass spectrometric analysis revealed that a number of Cj.-C _q dicarboxylic acids were contained in this fraction, said pimelic acid and azelaic acid prevailed quantitatively.

In comparative tests using samples of pure acids, _{any tyrosinase-inhibiting effect could be excluded for glutaric acid (C 5} ), while this effect is evident in compounds with 7 or more carbon atoms and is maximum in compounds with 9 and 11 carbon atoms.

In particular, enzymatic kinetic experiments showed that azelaic acid is a useful tyrosinase inhibitor with Ki = 4.10 ~ ⁴ M.

example

The in vivo use of the agents according to the invention is described in illustrated in the following experiments;

A cream based on azelaic acid (see the following Examples 3 to 5) is applied to the dark spots of the following patients for 30 days:

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Azelaic acid 15.0

Chlorocresol 0.1

Titanium dioxide - 1.0

Salicylic acid 2.0

Glycerol monostearate 2.0

Cetyl alcohol 3.0

Tween 80 5.0

Sodium lauryl ether sulfate 10.0

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(a) 20 patients suffering from chloasma,

(b) 3 patients suffering from poikiloderma according to Civatte,

(c) 1 patient suffering from delimited melanosis in the pre-cancerous stage according to Dubreuilh (malignant freckles) suffers.

After the end of the treatment, all patients showed a clear lightening of the hyperpxgmentated zones and in most cases an apparently complete cure with no side effects was achieved. After 5 months of observation none of the treated patients showed any traces of leukoderma.

It should be noted that precancerous circumscribed melanosis falls into the category of precancerous dermatoses, and it is of fundamental importance that there should be not only clinical cure in this patient but also revealed a histological cure.

Examples 3 to 5 are exemplary of the creams used in Example 2 above.

Example 3

Wt%

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Wt%

Ethanolamine lauryl ether sulfate 1.0

Olive oil 2.0

Vitamin C 1.0 [

make up to 100% by weight with distilled water

Example 4

The agent corresponds to that of Example 3, with the exception that 3% salicylic acid is used.

Example 5

The agent corresponds to that of Example 3, with the exception that 4% salicylic acid was used.

example

With regard to the treatment of melanoma, laboratory animals carried out the following tests:

Hardey-Passey melanoma samples were taken from infected rats and placed in small pieces in the subcutaneous tissue the left side of BaIb's C. Rats planted. the Animals were divided into two groups of 30 rats each, one of which by daily intraperitoneal Injections of saline suspension containing 2 mg alpha-mercaptoazelaic acid was treated.

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The control animals were given 0.1 ml of 0.9% saline solution; injected. Treatment was continued until death of the control | animals continued, then histological examinations were carried out both on the melanomas of the treated animals as ι

also carried out for the control animals. j

The following preliminary conclusions can be drawn from these experiments:

(a) The mean survival time of the animals treated with the substance containing azelaic acid, is extended;

(b) in the treated animals there is a substantial delay in the onset of melanoma development observed;

(c) in some of the animals treated, the melanomas are significantly reduced in size and extent;

(d) in other treated animals, instead of the

The formation of a black crust is observed in the tumor, i or no melanoma occurs at all; and J

i (c) finally were on histological examination extensive necrotic areas were found in the treated animals.

example

Human volunteers afflicted with malignant melanoma were treated orally with pure azelaic acid. This was given orally in a daily dose

of 10 "grams for a month with good tolerance administered.

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example

The following agent is suitable for use with subcutaneous injections of alphamercapto-azelaic acid:

Alpha-monomercapto-azelaic acid 2 mg Tween 80 5 mg

Vitamin C 1 mg

example

For further investigation of the inhibiting effect j of azelaic or dodecanedicarboxylic acid and their corresponding ^ Sodium salts on Harding-Passey melanomas in intraperitoneal, The following tests were carried out subcutaneously or orally:

Balb's C albino rats, which Harding-Passey melanomas transplanted were treated intraperitoneally, subcutaneously or orally with agents containing azelaic acid (C-9) or dodecanedicarboxylic acid (C-12) or its equivalent Contained sodium salts. Treatment was started 24 hours after the melanoma was inoculated. The Control animals were given daily intraperitoneally or subcutaneously 120 μΐ saline solution, which contained 1 mg of vitamin C and 2.5 mg Tween 80 was injected.

Each test animal was treated in one of the following ways: ^μ

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(a) Cq intraperitoneally (1):

Daily intraperitoneal injection of 120 μΐ saline solution containing 1 mg, azelaic acid, dissolved in 5 mg of its dimethyl ester, 1 mg of vitamin C and containing 2.5 mg Tween 80th

(b) Cg subcutaneous (1):

Daily subcutaneous injection of the agent described under (a) above.

(c) C. _? subcutaneous (1):

Daily subcutaneous injection of 120 μΐ saline solution, which 1 mg of dodecanedicarboxylic acid dissolved in 5 mg of its dimethyl ester, 1 mg of vitamin C and 2.5 mg of Tween 80 contains.

orally:

Oral administration ad libitum of an aqueous solution containing 20 mg of the sodium salt of azelaic acid and

Contains vitamin C per cm of water. ;

orally:

Oral administration ad libitum of an aqueous solution containing 20 mg of the sodium salt of dodecanedicarboxylic acid and contains 1 mg of vitamin C per cm of water.

The results of these tests are summarized in Table I below. It turns out that intraperitoneal, subcutaneous or oral administration of azelaic acid, dodecanedicarboxylic acid or their sodium salts to albino mice with transplanted Harding-Passey melanomas caused a significant delay in tumor growth.

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Tabel

Inhibiting effect of C _Q - and C _1. , - dicarboxylic acids on the growth of Harding-Passey melanomas

Balb's C albino mice

Vers. Number
the
animals

No.

links

Day of Occurrence Results at the end of the second week Results at the end of the 3rd week

Number of available dead of
Tumors

Size of the number of the present- size of the tumor by- killing of tumor- _f knife * tumor diameter

II

30th
30th

15th
15th
15th
15th
15th

control

C ₀ intraperi ^s toneal (1)

control
Cg subcutaneous (1) C ₉ oral (2) C ^ subcutaneous (1) (2)

7th

15. 8. 14. 15. 15. 15. 29/29

10-15 mm

15th 9/15 2-3 mm 1 14/14 10-15 um 3
***
9 7/12
4/6 2-4 mn
2-3 mm 2
***
7th 11/13
7/8 2-3 mn
2-4 mn

1
7th
2
1
2
0

29/29

11/14
7/7

10/10
4/5
9/11
7/8

15-20 mm

2-10 mm 15-20 mm 2-10 by 2-10 nm 2-10 mm 2-10 mm

Approximate size in vivo ** Size after death

The animals probably died because they did not drink for the first two days.

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Treatment was started 24 hours after inoculation.

Control: Daily intraperitoneal or subcutaneous injection of: 120 μΐ saline solution, 1 mg Vit. C, 2.5 mg Tween 80.

(1): Daily injection of 120 μΐ saline solution, 1 mg C or C - ^ - dicarboxylic acid (azelaic or dodecanedicarboxylic acid), dissolved in 5 mg of the corresponding dimethyl ester; 1 mg vitamin C, 2.5 mg Tween 80.

(2): Oral ad libitum: sodium salt of azelaic acid or sodium salt of dodecanedicarboxylic acid: 20 mg / cm H ₃ O; 1 mg vitamin C.

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Claims (21)

Claims 1. Means for the treatment of hyperpigmentary dermatoses or skin hyperpigmentations with at least one active ingredient in a pharmaceutically acceptable carrier, characterized in that there is at least one dicarboxylic acid with 7 to 13 carbon atoms as the active ingredient, a derivative thereof having at least one reducing functional group in the molecule or a contains pharmaceutically acceptable salt thereof. 2. Composition according to claim 1, characterized in that it i i in a form suitable for topical application is present. . ! 3. Means according to claims 1 or 2, characterized in that, that it contains a keratolytic agent, especially salicylic acid. 4. Composition according to claim 1, characterized in that it is in injectable form or in orally administrable form is present. 5. Agent according to one of claims 1 to 4, characterized in that that the reducing functional group is a mercapto group or groups acts. 809844/0824 original inspected M / 19 081 - J - 6. Composition according to Claims 1 to 4, characterized in that it contains azelaic acid as the active ingredient. 7. Composition according to Claims 1 to 4, characterized in that the active ingredient is a mercapto derivative the azelaic acid. 8. Composition according to Claims 1 to 4, characterized in that the active ingredient is a-Monomercaptoazelaic acid acts. 9. Means according to claims 1 to 4, characterized in that that the active ingredient is dimercaptoazelaic acid. 10. Composition according to one of claims 1 to 4, characterized in that it is pimelic acid, suberic acid, azelaic acid, sebacic acid, 1,9-Nonandi- ι carboxylic acid, 1,1O-decanedicarboxylic acid, 1,11-undecanedicarboxylic acid as the active ingredient and mono- or dimercapto extracts of these acids,; contains. ^: (11 bfono-a-mercapto derivative of suberic acid, azelaic acid, sebacic acid, j, 1, 9-nonanedicarboxylic acid, 1, 1 O-decanedicarboxylic acid and 1, 11 -Ündecandicarbonsäure. 12. Dimercapto derivative of a dicarboxylic acid having 7 to 13 carbon atoms. 8098U / Ö824 M / 19 081 13. The derivative of claim 12, wherein the dicarboxylic acid is selected is among pimelic acid, suberic acid, azelaic acid, sebacic acid / 1,9-nonanedicarboxylic acid, 1,10-decanedicarboxylic acid and 1,11-undecanedicarboxylic acid. 14. Process for the preparation of a mercapto derivative of a Dicarboxylic acid, characterized in that the dicarboxylic acid is used to introduce the desired number brominated by bromine atoms that are to be replaced by mercapto groups, the bromine derivative with thiourea reacted in a solvent to form the corresponding thiouronium salt and then the so formed Salt saponified to form the corresponding mercapto derivative. 15. The method according to claim 14, characterized in that a dicarboxylic acid having 7 to 13 carbon atoms is used. 16. The method according to claims 14 or 15, characterized in that the dicarboxylic acid is reacted with phosphorus pentachloride until the mixture is completely melted, adding anhydrous bromine to this melt with stirring while maintaining the temperature at 60 to 70 ⁰ C and then the bromine derivative isolated. 17. The method according to claims 14, 15 or 16, characterized in that the monobromo derivative is prepared by reacting 1 mol of acid with 50 g of PCl ₅ and then reacting the reaction product obtained with 60 ml of anhydrous bromine, the anhydrous 809844/082 * M / 1 & 081 Bromine added in small portions over approximately 6 hours. 18. The method according to claims 14, 15 or 16, characterized in that the dibromo derivative is prepared by reacting 1 mole of acid with 50 g of PCl ₅ and then reacting the product obtained with 120 ml of anhydrous bromine, the anhydrous bromine in the course of about 18 hours in small portions. 19. The method according to claim 14, characterized in that that one for the preparation of the Monomercaptoderxvats in the preparation stage of the alkylthiouronium salt Bringing 1 mol of monobromo derivative with 50 ml of ethyl alcohol and 1.1 mol of thiourea to implementation. 20. The method according to claim 14, characterized in that that one prepares the dibromide as an intermediate and then this with 2.2 mol of thiourea in 100 ml Ethyl alcohol reacts per mole of dibromide in the reaction mixture. 21. The method according to claims 14 to 20, characterized in that the saponification with sodium hydroxide performs. 8098U / 0824