DK169345B1 - Retinoic acid esters of erythromycin A, their preparation and pharmaceutical and cosmetic preparations thereof - Google Patents

Description

DK 169345 B1

The present invention relates to novel retinoic acid esters of erythromycin A, their preparation as well as pharmaceutical and cosmetic compositions containing them.

The retinoic acid esters are used in the treatment of various dermatoses, especially in the treatment of acne.

Furthermore, ongoing studies also show that the retinoic acid esters of this invention have antitumor effect.

The object of the present invention is primarily to provide novel retinoic acid esters for the treatment of infectious or non-infectious dermatoses which may be of bacterial or mycobacterial origin, and / or may be associated with the implantation of some yeasts which have a pathogenic character.

Acne is a polymorph (different lesion types occur in the same individual) skin disorder that occurs at puberty and which subsides spontaneously at 20-25 years of age in most cases.

In the affected individuals, acne occurs in areas rich in sebaceous glands, such as the forehead, face, sides 20 of the nose, chest and back, and this shows that this dermatosis has some dependence on sebum, the product synthesized by the gland.

There is no acne without seborrhea.

Although seborrhea is one of the consequences of the sudden onset of hormone levels that occur at puberty, acne does not appear to be associated with any hormonal disorder.

Although the etiopathogenesis of acne is not well defined, it begins with the formation of a characteristic lesion, the comedone. The latter is due to a clogging of the hair-fat channel as a result of a dekeratinization of the funnel-shaped region of the channel.

The main effect of this clogging is a change in the viscosity of the sebum and in the physical / chemical properties of the environment (pH, oxygen vapor pressure and the like).

This modification permits the hyper-proliferation of strains found in the skin, mainly Propionibacterium aenes, the anaerobic or air-tolerant strain.

Acne is by no means infectious in the sense that this dermatosis does not correspond to an implantation of a particular pathogenic strain and that it cannot be transmitted.

Finally, the bacterial hyper-proliferation results in a release into the environment of some proteases or hyaluronidases of bacterial origin that cause lysis of the follicle, and thus release of inflammatory compounds in the cutis, triggering an inflammatory reaction 10 in the organism.

Although the nature of the inflammatory compounds has not yet been determined, there seems to be little doubt about their bacterial origin, thus explaining the successful therapeutic effect of antibiotic compounds administered orally or locally in the treatment of inflammatory acne.

Among antibiotics, erythromycin and clindamycin are very frequently highlighted, but relatively high concentrations (especially in the case of erythromycin) are required to achieve a satisfactory effect.

As recent studies have also shown, some strains of Propionibacterium aenes develop a gradual resistance to erythromycin, lincomycin, and clindamycin, so treatment with these antibiotics may prove very effective.

Local use of clindamycin, and particularly of erythromycin, also poses a problem with penetration through the horn layer, thereby limiting their effectiveness.

The novel retinoic acid esters of erythromycin 30 A according to the invention provide a satisfactory solution to the problems associated with the use of this antibiotic. Studies have shown that these novel esters have a selective effect on the main organism responsible for the inflammation, ie. Propionibacterium aenes, 35 while having a very weak effect on skin organisms, such as Staphylococcus epidermidis, which allows skin diseases to be treated without disturbing the equilibrium of the skin.

It should also be noted that the novel retinoic acid esters, all-trans and 13-cis-retinoic acid esters of 5 erythromycin A, have been shown to be effective against strains of Propionibacterium aenes which are resistant to the parent antibiotic.

Furthermore, the novel retinoic acid esters of the invention have been found to be effective without the disadvantages of retinoic acid.

Thus, the novel esters are better tolerated by the skin and have been found to be much less toxic orally than the antibiotic / retinoic acid combination.

Compared to other known esters of antibiotics 15, the retinoic acid esters of the invention have the advantage of having a keratolytic effect in the case of the all-trans-retinoic acid ester and a potential antiseborrhea effect in the case of 13-cis. -retinoic acid ester, giving these esters an image as "prodrug".

The novel retinoic acid esters of erythromycin A according to the invention are more lipophilic, which makes it possible to improve penetration through the epidermis.

The known technique for the combination of retinoic acid and erythromycin is the product sold by 25 Cilag Laboratories under the name "Antibio-Aberel".

The prior art of esters of erythromycin A is represented by U.S. Patent No. 2,862,921, which relates to the preparation of saturated and monounsaturated fatty acid esters of erythromycin A such as erythromycin A monostearate and erythromycin A monooleate.

The retinoic acid esters of erythromycin A according to the invention have the following formula 35

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DK 169345 B1 4 H3c ^> ^ Sw, * 'CH3 5 HW> ° H V0H ^ CH3 H3C- ·] T' "CH3 R0V | Vc Hj CH3 CH3 V" tr 15 wherein R represents an all-trans-retinoyl group or a 13-cis-retinoyl group, the retinoyl group having the following formula ^ (II)

The present invention also relates to a process for the preparation of the all-trans and 13-cis-retinoic acid esters of erythromycin A, which is characterized by the characterizing part of claim 2.

30 Different esterification processes can be used.

The esterification is carried out by the process of the invention preferably in tetrahydrofuran alone or mixed with another organic solvent, such as pyridine, by reacting an excess of a mixed anhydride of all-trans or 13-cis-retinoic acid and carboxylic acid (produced in situ, f for example using ethyl chloroformate and all-trans (13-cis acid) with basic erythromycin A in the presence of an organic or inorganic base such as pyridine and / or sodium bicarbonate.

This method, using a mixed 5 anhydride, allows the formation of esters with retinoic acid at the 2 'position of erythromycin A without isomerization of the retinoyl group.

The present invention also relates to pharmaceutical compositions which may be administered topically, orally, parenterally or rectally, as well as cosmetic preparations for the treatment of various dermatoses, especially acne, these compositions being in anhydrous form and containing at least one all-trans or 13 -cis-retinoic acid ester of erythromycin A according to the invention, preferably at a concentration of between 0.01 and 10%, but more preferably between 0.05 and 1% by weight, based on the total weight of the composition.

For the preparation of the compositions of the invention containing as active ingredient at least one all-trans or 13-cis-retinoic acid ester of erythromycin A, carriers and adjuvants described in the pharmaceutical, cosmetic and related literature can be used.

For the preparation of solutions, e.g. one or more organic solvents are used which are acceptable from a physiological point of view.

In particular, the acceptable organic solvents are selected from acetone, isopropyl alcohol, fatty acid triglycerides, glycol ethers, C 1 -C 4 alkyl esters of short chain acids and polytetrahydrofuran ethers.

The compositions of the invention may also contain thickeners such as cellulose and / or cellulose derivatives at a concentration of 0.5-20% by weight, based on the total weight of the composition.

The compositions of the invention may further contain at least one other known anti-acne agent in combination DK 169345 B1 6 with at least one retinoic acid ester of Erythromycin A according to the invention.

A common adjuvant selected from antioxidants, preservatives, perfumes and dyes can be added if needed.

5

Among the antioxidants which can be used, e.g. mention is made of tert.butyl hydroxyquinone, butyl hydroxyanisole, butyl hydroxytoluene and α-tocopherol and its derivatives.

The conversions of the compounds of the invention into pharmacological and galenic forms are carried out in a known manner.

The galenic forms for local use may be creams, milk, gels, more or less thickened lotions, lotions contained in tampons, ointments or sticks or aerosol preparations in the form of spray or foam.

The compositions for oral use may be in the form of tablets, capsules, dragees, syrups, suspensions, emulsions, powders, granules or solutions. The oral dose is approx. 0.1-5 mg / kg per per day, and preferably 1-2.5 mg / kg per day. day.

The preparations may also be in the form of suppositories.

The treatment of acne using compositions of the invention for topical application consists of applying a sufficient amount of two to three times a day to the areas of the skin to be treated, for a period of 6-30 weeks, and preferably 12 -24 weeks.

The compositions of the invention can also be used preventively, i.e. on skin areas that are susceptible to acne attacks.

Examination of the effect of retinoic esters of erythromycin A.

The effect of retinoic acid esters of erythromycin A is investigated by the dilution method to determine the minimum inhibitory concentration (MIC), a method described in 35 and used by G.A. Denys et al., Antimicrobial Agents and Chemotherapy (1983) 23, pp. 335-337, and J.J. Leyden et al., DK 169345 B1 7 J. Am. Acad. Dermatol. (1983) 8, (1) pp. 41-45, using as the Propionibacterium acnes strain, strain P37, supplied by Cunliffe and the Netherlands.

This strain P37 has been the subject of studies 5 described in the following publications: - J. Greenman, K.T. Holland and W.J. Cunliffe,

Journal of General Microbiology (1983) 129, pp. 1301-1307, - E. Ingham, K.T. Holland, G. Gowland and W.J.

Cunliffe, ibid (1980) 118, pp. 59-65 and 10 - K.T. Holland, J. Greenman and W.J. Cunliffe, Journal of Applied Bacteriology (1979) £ 7, pp. 383-394.

Selection and isolation of sensitive and resistant populations.

Strain P37 is sensitive to erythromycin as shown at the minimum inhibitory concentration (MIC = 0.78 µg / ml).

In contrast, after 8 successive subcultures in the same medium (RCMX 19/20 and DMSO 19/20 after volume), in order to obtain a gradual adaptation of the strain to this medium, gradual resistance to erythromycin in the following form is shown. :

After distributing a standard inoculum (OD = 1.8 at 450 nm) on agar ("RCM" + furazolidone) in a Petri dish, a 9 mm diameter disc is placed in the center 25 of the dish. 50 µg Erythromycin (dissolved in DMSO) is deposited on the disc.

After 6 days at 36 ° C in an anaerobic medium (BBL Gas-Pak system), a zone over which the growth of the strain is inhibited is clearly visible (total diameter = 42 mm), with the majority of the colonies being in the periphery of inhibition zone.

In contrast, some colonies are clearly seen within this zone.

The two types of colonies are then removed by scraping the agar medium (sterilized platinum eye): x Reinforced Clostridium Medium ("OXOID")

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DK 169345 B1 8 Θ 1) strains designated P37 E due to their apparent resistance to erythromycin are withdrawn from within the inhibition zone and 2) strains designated P37 E are withdrawn from an area of 5 1 cm beyond the periphery of the inhibition zone.

Φ

After isolation and culture, strains P37 E Θ and P37 E also show very different sensitivities to erythromycin as illustrated by the following respective MIC values: 10 MIC (pg / ml) P 37 0.78 P 37 E® 0.78 P 37 Εθ 50 15 This phenomenon is confirmed by examination of IC IC q (50% inhibitory concentration), which represents the erythromycin concentration at which 50% of surviving organisms are found among the population at a constant culture period.

20 IC5Q (pg / ml) P 37 50 P 37 E® 5 P 37 E® 100 25 The minimum inhibitory concentration (MIC) expressed as µg / ml of the retinoic acid esters of erythromycin A tested against strains P 37® and P 37® is listed in the following table. 1 35

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DK 169345 B1 9

Retinoic acid esters P 37 ΕΦ P 37 Εθ according to the invention (sensitive) (resistant) 2'-0- (all-trans-retinoyl) -erythromycin A 14 13 5 2'-0- (13-cis-retinoyl) -erythromycin A 20 34

Check;

2'-O-oleoylerythromycin A

(2-9) 50 100 3-O-oleoylline comycin 19 42 3-O-oleoyl-clindamycin 54> 138

Erythromycin A 1> 50

Lineomycin 13 66 15

Clindamycin 1 10

The table below shows the minimum inhibitory concentrations of the retinoic acid esters of the invention for two strains of Staphylococcus epidermidis:

Retinoic acid esters of the invention Staph »epi. 3 Staph. epi. 6 2'-O- (all-trans-retinoyl) -25-erythromycin A 75 80 21-O- (13-cis-retinoyl) -erythromycin A 110 110

Check; 30 - erythromycin A 13 30 clindamycin 7 8 lincomycin 14 20 35 DK 169345 B1 10

The strain "Staph, epi. 3" is isolated from a patient carrying acne, whereas the strain "Staph, epi. 6" is isolated from a patient not carrying acne. The isolation of these strains is done by the method of Williamson and Kligman 5 ("A new method for the quantitative investigation of cutaneous bacteria", P. Williamson and A. Kligman, J.I.D.,

Vol. 45, No. 6, 1965). Decimal dilutions of the samples are made and a selective medium enabling Staphylococcus to be isolated is inoculated with 0.1 ml of these dilutions.

As can be seen from the first table, the retinoic acid esters of erythromycin A are more effective against the resistant strains of Propionibacterium aenes than the starting antibiotic. Furthermore, the 2'-oleic acid ester of erythromycin A (U.S. Patent No. 2S62,921) and the 3-oleic acid ester of clindamycin (German Publication No. 2,017,003), taken as comparative esters, are found to be substantially less active over for the sensitive (P 37 E +) and the resistant (P 37 E ") strains than the esters of the invention, thus emphasizing the value of the retinoic acid esters of erythromycin A. The second table, in turn, shows the value of these retinoic acid esters of erythromycin A with respect to to "skin ecology" in that they are much less effective against Staphylococcus epidermidis strains than the starting antibiotic.

More examples of the preparation of retinoic acid esters of the invention and more examples of pharmaceutical or cosmetic compositions for use in the treatment of dermatoses, especially acne, will be given below.

30

Example 1

Preparation of 2'-O- (13-cis-retinoyl) -erythromycin A

5 g (16.6 mmol) of 13-cis-retinoic acid are dissolved in 35 ml of anhydrous tetrahydrofuran in a round bottom flask under an inert atmosphere. The reaction mixture is cooled to 0 ° C and 3 ml (38 mmol) of anhydrous pyridine and 1.6 ml (16.6 mmol) of ethyl chloroformate are then poured into the mixture. The solution is stirred for 5 minutes and 2.5 g (30 mmol) of sodium bicarbonate are added and then 4.9 g (6.7 mmol) of erythromycin A, which is already dissolved in 150 ml of tetrahydrofuran. The reaction mixture is then stirred for 10 hours, allowing the temperature to rise to ambient temperature (thin layer chromatography on silica gel: methylene chloride / methanol, 10%). The solution is then poured into 60 ml of water and then extracted with ethyl acetate. The organic phase is dried over magnesium sulfate, filtered and then concentrated under partial vacuum. The crude product thus formed is chromatographed on a silica gel column (high pressure liquid chromatography, hereafter abbreviated HPLC), using a mixture of ethyl acetate and hexane (7: 3) as the eluent, resulting in isolation of 4.4 g (65% yield). ) purely 2'-0-

Q

- (13-cis-retinoyl) -erythromycin A. Melting point: 82 C (hexane / ethyl acetate).

[α] 25 = -17 ° (C = 6 mg / ml dichloromethane).

Microanalysis: M = 1016.4 C% H% N%

Calcd: 67.36 9.22 1.38

Found: 67.48 9.32 1.38

Infrared Spectroscopy: band at 1735 cm -1 1 (ester) NMR (CDCl3, internal reference TMS).

The negative γ effects at positions 1 '(-2.2 ppm) and 3' (-2.1 ppm) show the 2 'position of the ester. The carbon atoms C "20 (20.94 ppm), C" 14 (117.28 ppm) and C "12 d · 31'® PPm) of the retinoic acid chain are consistent with the 13-cis stereochemistry of the retinoic acid chain.

30

Example 2

Preparation of 21-O- (all-trans-retinoyl) -erythromycin A

5 g (16.6 mmol) of all-trans-retinoic acid are dissolved in 35 ml of anhydrous tetrahydrofuran in a round bottom flask under an inert atmosphere. The reaction mixture is cooled to 0 ° C and 3 ml (38 mmol) of anhydrous pyridine and 1.6 ml (16.6 mmol) of DK 169345 B1

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12 ethyl chloroformate is then poured into the mixture. The solution is stirred for 5 minutes and 2.5 g (30 mmol) of sodium bicarbonate are added and then 4.9 g (6.7 mmol) of erythromycin A, which is already dissolved in 150 ml of tetrahydrofuran.

The reaction mixture is then stirred for 10 hours, allowing the temperature to rise to ambient temperature (thin layer chromatography on silica gel: methylene chloride / methanol, 10%). The solution is poured into 60 ml of water and then extracted with ethyl acetate. The organic phase is dried over magnesium sulfate, filtered and then concentrated under par * -tile vacuum. The crude product thus formed is chromatographed on a silica gel column (HPLC) using a mixture of ethyl acetate and hexane (7.:3) as the eluent, resulting in isolation of 4.1 g (60% yield) of pure 2'-0. - (all-trans-retinoyl) -erythromycin A. Melting point = 76 ° C (ethyl acetate / hexane).

[α] D = -65 ° (C = 2 mg / ml dichloromethane).

Microanalysis: C57 H93 NO4 * 4ii20i = 1088.5 C% H% N% 20 Calculated: 62.89 9.35 1.29

Found: 62.91 8.90 1.29.

13 C NMR (CDCl 3, internal reference TMS).

The negative γ effects at positions 1 '(-2 ppm) and 3' (-1.9 ppm) show the 2 'position of the ester. The carbon atoms 25 C "20 PPm)" c "14 (II9 / 36 ppm) and C" 12 (135.19 ppm) are consistent with the all-trans stereochemistry of the retinoic acid chain.

30 35

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13 DK 169345 B1

Pharmaceutical and Cosmetic Preparations A. Gels for Local Acne Treatment 1. Hydroxypropyl Cellulose 1 g

Butyl hydroxytoluene 0.05 g of 5-O- (13-cis-retinoyl) lincomycin 0.5 g

Isopropanol, up to 100 g 2. Hydroxypropyl cellulose 1.5 g

Butyl hydroxytoluene 0.05 g 3-O- (all-trans-retinoyl) clindamycin 0.3 g

Isopropanol, up to 100 g B. Lotions for local treatment of acne 1. Butyl hydroxytoluene 0.05 g 21-0- (all-trans-retinoyl) -erythromycin A 1 g 15

Cg-C ^ fatty acid triglycerides, up to 100 g

The active compound in this example can be replaced by the same amount of 2'-O- (13-cis-retinoyl) -erythromycin A.

2. Butyl hydroxytoluene 0.05 g 3-O- (13-cis-retinoyl) clindamycin 0.7 g

Polytetrahydrofuran dimethyl ether (viscosity 22 cP) of the formula CH3 0- £ (CH 2) 2 “C® 2 2 3 η 25 wherein η = '5, up to 100 g C. Pin for local treatment of acne

White Vaseline 52 g

Vaseline oil 15 g 30

Refined paraffin 32 g 2'-O- (all-trans-retinoyl) -erythromycin A 1 g D. Suppository (1 unit preparation) 2'-O- (all-trans-retinoyl) -erythromycin A 0.05 g

Cg-C2-phthalic acid triglycerides 0.25 g 35 Semi-synthetic glycerides, up to 2 g 14 Capsules of 500 mg

The capsule walls consist of glycerine, sorbitol and gelatin.

1. Capsule with 50 mg of active compound 5 2'-O- (13-cis-retinoyl) -erythromycin A 50 mg

Liquid Vaseline Oil 200 mg

Thickened petroleum jelly 250 mg 2. Capsule with 10 mg active compound 10 2'-O- (13-cis-retinoyl) -erythromycin A 10 mg

Butyl hydroxyaminosis 0.05 mg

Butyl hydroxytoluene 0.05 mg

Glycerol Tribhenate 100 mg

Cr “Ci9 fatty acid triglycerides, up to 500 mg 15 0 z F. Capsules

The capsule walls consist of gelatin and titanium dioxide.

21-O- (all-trans-retinoyl) -erythromycin A 20 mg 20

Colloidal silica 2 mg

Magnesium stearate 2 mg

Corn starch * 76 mg

Lactose, up to 250 mg 25 30 35

Claims (6)

1. All-trans and 13-cis-retinoic acid esters of erythromycin A of formula I 5. CH3 H (4 ^ J> 0H 0H ^ CH3 H3C '' '|' * CH3 R0 \ r / N ^ CH3 "Λ" JUL ^ mj 'p and 3 CH, IJ CH- wherein R is an all-trans-retinoyl group or a 13-20 -cis-retinoyl group, the retinoyl group having the following formula 25. dl) Γ <£) <E> or 30 mixtures thereof and salts of said esters. Process for the preparation of the all-trans and 13-cis-retinoic acid esters of erythromycin A according to claim 1, characterized in that an excess of a mixed anhydride of all-trans or 13-cis-retinoic acid in an anhydrous DK 169345 B1 16 organic solvent medium is reacted with erythromycin A in basic form in the presence of an organic or inorganic base. Pharmaceutical and cosmetic compositions for the treatment of various dermatoses, especially acne, characterized in that in an anhydrous carrier they contain at least one all-trans or 13-cis-retinoic acid ester of erythromycin A according to claim first Composition according to claim 3, characterized in that it contains from 0.01 to 10% by weight, and preferably from 0.05 to 1% of active compound. Composition according to any one of claims 3 to 4, characterized in that the carrier material is acetone, isopropyl alcohol, fatty acid triglycerides, glycol ethers, C Composition according to any one of claims 3-5, characterized in that it further contains a thickening agent such as cellulose and / or cellulose derivatives 20. an amount of 0.5-20% by weight, based on the total weight of the composition.