CH685674A5 - Ultramicroemulsions spontaneously dispersible concentrates with pharmacologically active esters of Apocarotinolen. - Google Patents

Description

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

description

INTRODUCTION

The present invention relates to ultramicroemulsions from spontaneously dispersible concentrates with esters of apocarotenols, esters with apocarotenols, processes for their preparation, and the use of the spontaneously dispersible concentrates containing apocarotinyl esters as agents for producing a pharmaceutical preparation having activity against tumors, eczema and psoriasis.

Selected esters of apocarotenols surprisingly have a very good effect against tumors, psoriasis and eczema, especially when they have been incorporated into spontaneously dispersible concentrates which, when diluted with water, give rise to thermodynamically stable ultramicroemulsions with micelles with a hydrodynamic radius of 1.5 to 3 nm.

DESCRIPTION OF THE INVENTION

The esters with apocarotenols selected for the present invention have the formulas (I) to (IX):

o ch2 — o — cr,

(I)

—Ri

(ii)

2nd

CH 685 674 A5

10th

15

20th

O

II

ch2 — o — c - r

(Ml)

ch2 — o— c — rl

(IV)

25th

30th

(V)

35

40

ff

CHj-O C-R,

45

50

55

(VI)

I.

ch2 — o —- c — r,

(VU)

60

65

3rd

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

O

II

ch2 — o — c — r1

(VIII)

cho

(IX)

where in the formulas (I) to (IX) Ri a Cr to C32 alkyl, a C2 to C32 alkenyl or alkapolyen, a C2 to C32 alkynyl group or a radical of the formulas (X ) or (XI) represents:

?

H,

r2 - (- c h == c h-c = c h-)

n

(x)

Ç ":

- (- ch = ch-c = ch -) - {- ch = ch-ch- (j -) - ch = ch

ch,

(XI)

where n denotes the numbers 1, 2, 3, 4 or 5 and R2 denotes one of the radicals of the formulas

4th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

stands.

The alkyl, alkenyl or alkapolyene groups (i.e. the corresponding alkadienes, alkatrienes, alkatatenes, alkapentaenes, alkahexaenes or alkapentaenes) and the alkynyl groups in R 1 can be straight-chain or branched.

Preferred compounds of the formulas (I) to (IX) are those in which R 1 is a C 1 -C 4 -alkyl, a C 2 -C 8 -alkenyl or alkapolyen group, a C 2 -C -alkynyl group or a radical of the formulas (XII) and (XIII) denote:

(XII)

5

5

10th

15

20th

25th

30th

35

40

45

50

55

60

CH 685 674 A5

ch3

(XIII)

Compounds of the formulas (I) to (IX) are particularly important, in which R 1 is a C4 to C is alkyl, a C2 to cis alkenyl or alkapolyen, a C2 to Cis alkynyl group or Represents radical of formula (XII).

The compounds of the formulas (I) to (IX) can exist in various stereoisomeric or rotational forms. Examples of compounds of the formulas (I) to (IX) include:

8'-Apo-β-carotin-3-ol-10-undecenoate

8'-Apo-β-carotene-3,8'-diol-di-10-undecenoate

3 ', 4'-Dihydro-2'-apo-β-carotin-2-ol-10-undecenoate

12'-Apo-β-carotene-12'-ol-10-undecenoate

10'-apo-β-carotene-10'-ol-10-undecenoate

8'-Apo-β-carotene-8'-ol-10-undecenoate

8'-apo-β-carotene-8'-ol laurate

8-apo-β-carotene-8'-ol palmitate

8-apo-β-carotin-8'-ol-10 stearate

6'-Apo-β-carotene-6'-oi-10-undecenoate

Azafrinyl 10 undecenoate

Azafrinyl Iaurate

Azafrinyl palmitate

Azafrinyl stearate

The compounds of the formulas (I) to (IX) can generally be prepared by the following known processes a) or b):

a) implementation of a compound of formula

R, COOH

wherein Ri represents a Cr to C32 alkyl, a C2 to C32 alkenyl, a C2 to C32 alkapolyen, a C2 to C32 alkynyl group or a radical of the formulas (X), or (XI ) is, with N, N'-carbonyldiimidazole at 0 to 50 ° C with a protective gas supply and addition of a catalytic amount of an alcoholate in an inert solvent and subsequent alcoholysis of the imidazolides formed with a compound of formulas (XV) to (XXIII):

(XV)

12'-apo-β-carotin-12'-ol

Total synthesis: R. Rüegg et al., Helv. Chim. Acta 42, 847-864 (1959)

6

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

ch2oh

(XVI)

10-apo-ß-carotin-10'-ol

Total synthesis: R. Rüegg et al., Helv. Chim. Acta 42, 847-864 (1959)

(XVII)

8'-apo-p-carotene-8'-ol

Total synthesis: R. Rüegg et al., Helv. Chim. Acta 42, 847-864 (1959)

(XVIII)

6'-apo-β-carotin-6'-ol

Total synthesis: R. Rüegg et al., Helv. Chim. Acta 42, 847-864 (1959)

(XIX)

4'-apo-β-carotin-4'-ol

Total synthesis: R. Rüegg et al., Helv. Chim. Acta 42, 847-864 (1959)

7

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

(XX)

2'-apo-β-carotin-2'-ol

Total synthesis: R. Rüegg et al., Helv. Chim. Acta 42.847-864 (1959)

(XXI)

Azafrinol; (5R, 6R) -5,6-dihydroxy-5,6-dihydro-10'-apo-ß-carotin-10'-ol

Total synthesis: W. Eschenmoser and C.H. Eugster, Helv. Chim. Acta gl, 822 (1978). Merck index 11,911

(XXIi)

ß-citraurol; ß-Citraurinol

Synthesis: H. Pfander et al., Helv. Chim. Acta 22, 716-727, (1980)

(XXIII)

ß-citraurin; ß-Citraurinin, Apo-2-Zeaxanthinal, (3R) -3-Hydroxy-8'-apo-ß-carotin-8'-al Synthesis: H. Pfander et al., Helv. Chim. Acta SS. 716, 1377 (1980). Merck index 11.2326.

8th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

b) formation of the chloride or bromide of a compound of the formula r., cooh in which R3 is a Ci to C32 alkyl, a C2 to C32 alkenyl, a C2 to C32 alkapolyen, a C2 to C32-alkynyl group or a radical of the formulas (X) or (XI), with a chlorinating or brominating agent, such as, for example Thionyl chloride, oxalyl chloride, and subsequent reaction with a compound of the formulas (XV) to (XXIII) at a temperature of 0 to 50 ° C with supply of protective gas, in an inert solvent, such as e.g. Toluene or tetrahydrofuran, and in the presence of a catalyst such as e.g. Pyridine, dimethylformamide or p-dimethylaminopyridine.

The procedures c) and d) are new.

c) Production of fatty acid esters of 8'-apo-ß-carotin-8'-ol by the DIBAH process.

The compound (XVII), i.e. the C3o alcohol:

(XVII)

has so far been produced by reduction of 8'-apo-ß-carotin-8'-al with LiAIH4 in ether. [R. Rüegg, M. Montavon, G. Ryser, G. Saucy, G. Schwieter and O. Isler, Helv. Chim. Acta 1959, 42, 854]. The C30 alcohol was characterized as follows: unstable orange crystals with mp 148-149 ° C; /.max (petroleum ether): 426, 453 nm; O-acetyl compound: orange flake, mp 130-132 ° C, Àmax (petroleum ether): 426, 452 nm; easy elimination reaction in chromatography experiments on AI2O3. Other, especially higher esters have not yet been described.

The new, economical production process for the C3o alcohol leads to the reduction of the 8'-apo-ß-ca-rotin-8'-acidic acid ester by means of DIBAH (diisobutylaluminium hydride).

Can alcohol. 8'-apo-β-carotene-8'-ol

3.0 g of methyl 8'-apo-β-carotene-8'-oat and 300 ml of absolute diethyl ether are placed in a dry 1 liter three-necked flask with an inlet tube for protective gas, cooler, magnetic stirrer and septum. The mixture is then cooled to -30 ° C. under N2 and 3.0 ml of an approximately 4.5 molar solution of DIBAH in hexane are then slowly added dropwise using a syringe. There is no visible reaction. Now the mixture is slowly brought to RT and the completeness of the reduction is checked using UV / VIS spectra and DC. If all the esters have not yet been reduced, the mixture is cooled again to -30 ° C. and the required amount of DIBAH is added; etc. An excess of DIBAH considerably reduces the achievable yield of C30-Al-alcohol.

For working up, the mixture is cooled to 0 ° C., then mixed with 5 ml of ethyl acetate and finally carefully mixed with small pieces of ice until no more foaming occurs. The cooling bath is then removed, the mixture is mixed with Celite and then suction filtered. The filter cake is washed with Et20 / AcOEt / isopropanol 10: 4: 1 dye-free and the clear, red filtrate i.V. evaporated to dryness. The already crystalline, red residue can be recrystallized either from hot toluene / hexane or ethyl acetate / hexane.

Yield of C3o alcohol 2.2 to 2.4 g (80-90%). Orange-red needles, mp 148-149 ° C; UV / VIS (Et20, qualitative): Xmax: 416, 425, 452 nm; IR (KBr): 2.77 (3603 crrri, OH free), no bands in the car

9

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

bonyl area; CI-MS (NH3): 419 [M + 1, (16)} 401 [M + 1-H20, (100)], 402 [M + 2-H20, (30)], 309 [M + 1-To -Ilio I, (5%)]

Preparation and characterization of Undec-10-enoic acid -r8-Aoo-B-carotin-8'-vl-ester1

A solution of 400 mg C3o alcohol in 20 ml abs. EkO and 1.5 ml abs. Pyridine is cooled in a three-necked flask with a magnetic stirrer, N2 inlet tube, reflux condenser and septum under N2 to minus 30 ° C. and 200 ml of undec-10-enoyl chloride are then added dropwise while stirring. When the addition is complete, the temperature is allowed to rise slowly to 20 ° C. and the completeness of the esterification is checked by TLC on silica gel plates with the eluent benzene / petroleum ether 1: 1. The ester has a significantly higher Rf. value than the alcohol. Now the mixture i.V. evaporated, the residue taken up in benzene and the solution quickly filtered through a short column of CaC03 (Merck Art. 2066). The filtrate is evaporated and the residue is chromatographed on 40 g of silica gel (Merck, 15-40 (im) with benzene. The substance from the red zone is collected, evaporated in vacuo and the ester obtained from t-butyl methyl ether / pentane at -20 ° C. 450 mg of light red crystals are obtained after drying at 40 ° C./0.01 Torr, yield 80%, mp 76-77 ° C., after sintering from 75 ° C.

The esters were prepared in an analogous manner with lauric acid, palmitic acid and stearic acid chloride. Analytical data below.

d) Production of azafrinol and azafrinylesters

Azafrinol

The connection

Azafrinol; (5R, 6R) -5,6-Dihydro-10'-apo-8-ß-carotene-5,6,10'-triol) was first produced by W. Eschenmoser and CH Eugster by reduction of azafrin methyl ester and as an unstable oil described; s. Helv. Chim. Acta 1975, § £, 1722.

Through a modified reduction and work-up, azafrinol can be isolated as a well-crystallized compound and then also characterized in detail. Azafrinol is thus obtained by crystallization from benzene / hexane in bright light orange-yellow needles. The compound is particularly sensitive to acids and loses water as a result of ring splitting. This reaction already occurs during chromatography on silica gel.

Manufacturing and characterization

In a carefully dried three-necked flask, equipped with a reflux condenser, gas inlet pipe, sep

h2oh

(XXI)

10th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

Tum and magnetic stirrer are poured over 3 g of azafrin methyl ester with 300 ml of absolute ether and then cooled to -30 ° C. under protective gas and with good stirring. Then 3.5 ml of an approximately 4.5 molar solution of DIBAH (diisobutylaluminium hydride) in hexane are added dropwise through the septum by means of a syringe; there is no visible reaction. During a slow temperature increase to 0 ° C, the course of the reaction is checked using UVA / IS spectra and TLC.

If the reduction is still incomplete, the mixture is cooled again to -30 ° C. and a corresponding amount of DIBAH solution is added. A small excess of DIBAH does not have a significant impact on the yield, but a larger excess must be avoided.

After the reaction has ended, small pieces of ice are added to the solution very carefully until foaming no longer occurs and the precipitation of aluminum hydroxide has ended. After adding enough Celite and 0.2 ml of ethyldiisopropylamine, the product is filtered off with suction and washed with a mixture of ether / isopropyl alcohol 5: 1 without dye. The yellow-orange colored filtrate is then i.V. evaporated and the crystalline residue dissolved hot in toluene / isopropyl alcohol 99: 1 and, in the case of turbidity, the solution was quickly filtered through a small, wide column of CaC03. The filtrate is evaporated again and the residue is recrystallized from hot benzene / hexane or ethyl acetate / hexane. After drying at 40 ° C / 0.01 Torr, 2.2 g of bright bright /

orange needles. Mp 133-134 ° C (vacuum tube). See the analytical data reported below.

Production of undec-10-enoic acid [azafrinyl] ester

180 mg of freshly recrystallised azafrinol under N2 in 15 ml of EtïO and 0.5 ml of abs.

Dissolved pyridine and then cooled to minus 30 ° C. Then 100 ml of 10-unden-oyl chloride are added dropwise with a syringe. After stirring for one hour at a temperature of minus 30 ° C., the mixture is allowed to come to RT. The mixture is then i.V. evaporated, the residue taken up in hexane / ether 3: 1 and the solubles through a short and wide column of CaCC> 3 (Merck, Art. 2066)

filtered. The lemon-yellow eluate is evaporated again and chromatographed on a column of Sephadex LH-20 (swollen in the same solvent) using Et20 / hexane / isopropanol 50: 49: 1 + a trace of ethyldiisopropylamine. 161 mg of vacuum-dry, viscous oil were obtained from the main orange zone, which gradually crystallized. No significant m.p. could be obtained. UVA / IS (Et2Ö,

qual.): sharp three-band spectrum with Xmax 375, 394.5, 419 nm; (Qual. Benzene): 377, 402, 428 nm.

in an analogous manner, the esters n = 9; n = 13; n = 15, or

Azafrinyl laurate, azafrinyl palmitate and azafrinyl stearate. Analytical data are summarized below.

The apocarotenes of the formulas (I) to (IX) surprisingly have an excellent antitumor effect, especially when they have been incorporated into spontaneously dispersible concentrates which, when diluted with water, give rise to thermodynamically stable ultramicroemulsions with a hydrodynamic radius of 1.5 to 3 nm . (Measurements, against water, with dynamic, quasi-elastic light scattering ÖLS). The present invention therefore essentially also relates to spontaneously dispersible concentrates with the apocarotinyl esters of the formulas (I) to (IX).

The apocarotenes of the formulas (I) to (IX) are almost water-insoluble and polymer agglomerated compounds. However, in order that the molecules of these compounds diffuse through the membrane barrier of the tumor cells and become effective inside the cell, such compounds first have to be solubilized in a suitable manner in the aqueous medium. By means of the formation of thermodynamically stable oil-in-water ultramicroemulsions with the help of specially selected co-surfactants or solubilizers on the one hand and suitable surfactants on the other hand, an optimal degree of solubilization of the apocarotinyl esters can be achieved.

All experimental observations on ultramicroemulsions designed in this way can be interpreted uniformly by the assumption that the selected surfactants and cosurfactants, taken as a balanced system, form organized aggregates, so-called micelles, in the aqueous phase. They are more or less spherical in shape, with a hydrodynamic radius of 1.5 to 3 nm.

3rd

n ^ CH3

Oh

11

5

10th

15

20th

25th

30th

35

40

45

50

55

60

CH 685 674 A5

The surfactants and hydrotropes (cosurfactants) allow a boundary layer to form between the outer, aqueous phase and the inner, oily phase of the microemulsion [containing the active pharmaceutical ingredients dissolved in the bio-surfactant solubilizer], as a result of which the mixing of these two phases is avoided. In the oily, inner phase, the active substance molecules are in monomeric or oligomeric agglomerated form.

The micelles of the ultramicroemulsions according to the invention with the active substance-containing inner phase are provided with a surfactant coat at their boundary layer, which makes them easy to diffuse through the cell membrane into the interior of the cell. The diffusion through the plasma membrane of the cells takes place exclusively due to thermal molecular movements.

The direction that a specific diffusion process takes is determined by the difference in concentration that exists on the plasma membrane between outside and inside the individual cell. The diffusion continues along the concentration gradient until it is broken down. The concentration of active substance or of the active substance system (“multiple drug system”) is balanced between the extracellular zone and the interior of the individual cell, whereby delayed release effects (“slow release effects”) can also occur. Such diffusion processes take place independently of any energy supply. They have no relation to cellular metabolic energy.

The diffusion speed obeys Fick's law for diffusion processes in the direction of a concentration gradient:

- - = -D - EQUATION (A)

dt q dx where dm is the amount in moles of the active substance molecules penetrating a cell membrane surface q (in cm2) per time dt (in seconds). D is the diffusion coefficient and de is the concentration difference over the distance dx.

According to Nernst, the diffusion coefficient depends on the absolute temperature and the frictional resistance f

R T kT

D = - - = EQUATION (B)

N f f

The frictional resistance depends on Stoke's law f = 6 re -n r EQUALIZATION (C)

on the viscosity of the diffusing solution and on the radius of the diffusing particles. Substituting f with 6 rj r in the Nernst equation gives the Sutherland-Einstein equation for the diffusion coefficient

RT 1 kT

D = = EQUATION (D)

N 6 it i] r 6 each ri r where k represents the Boltzmann constant.

If there is a uniform drop in concentration in the membrane of the

dc Ac morzelle assumed, the expression in the diffusion law can be replaced by

dx x den. (Concentration difference ac over the cell membrane of thickness x). x is a constant size for a certain cell membrane, which is why it can be combined with the diffusion coefficient to form a new constant, the permeability coefficient P.

D

P = - EQUATION (E)

X

12th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

dm 1

The expression - in the diffusion equation the flux is called J and has the dimensions

dt q on moles per second per cm2. The negative sign on the right side of the diffusion equation indicates that the transport of the active substance molecules or the active substance system takes place in the direction away from the decreasing concentration.

So it is

■ RT 1 A kT 1

J = -Päc = - ~ - Ac = - Ac

Nx6 jc ri r x 6 n ti r equation (f)

From this equation it follows that the speed of the diffusion process or the strength of the active substance transport through the membrane of the tumor cell is determined:

1. the difference in concentration Ac in the two compartments

2. of the particle radius of the diffusing drug molecule or drug system

3. the viscosity of the diffusing aqueous solution (emulsion)

4. on the temperature.

The concentrates which are spontaneously dispersible according to the invention contain:

0.001 to 30% by weight of individual esters of apocarotenols of the formulas (I) to (IX), or combinations of such esters, and

0 to 40% by weight of a pharmaceutically compatible solvent or solvent mixture serving as a hydrotrope or co-emulsifier 0.001 to 90% by weight of a pharmaceutically compatible surfactant or surfactant mixture

0 to 10% by weight of a vitamin or provitamin

0 to 10% by weight of a penetration enhancer, a stabilizer or radical scavenger and conventional carriers and / or diluents.

The surfactants or surfactant mixtures to be used according to the invention can be anionic, cationic, amphoteric or non-ionic. They are preferably amphoteric or non-ionic and have an HLB ratio (i.e. a “hydrophilic-lipophilic balance”) between 2 and 18; it is preferably between 2 and 6 on the one hand and 10 and 15 on the other. HLB values provide information about the hydrophilic and lipophilic properties of an emulsifier. See also "Hydrophile-Lipophile Balance: History and recent Developments" by Paul Becher in the Journal of Dispersion Science and Technology 5 (1), 81-96 (1984).

Suitable anionic surfactants can be both so-called water-soluble soaps and water-soluble synthetic compounds.

Suitable soaps are the alkali, alkaline earth or optionally substituted ammonium salts of higher fatty acids (C-io to C22), e.g. the natural Na or K salts of oleic or stearic acid, or of natural fatty acid mixtures, which among other things Made from coconut or tallow oil. The surfactants also include the fatty acid methyl taurine salts and modified and unmodified phospholipids.

However, so-called synthetic surfactants are used more frequently, in particular fatty sulfonates, fatty sulfates, sulfonated benzimidazole derivatives or alkylarylsulfonates.

The fatty sulfonates and sulfates are generally present as alkali, alkaline earth or optionally substituted ammonium salts and generally have an alkyl radical having 8 to 22 carbon atoms, alkyl also including the alkyl part of acyl radicals. Examples of these are the Na or Ca salt of Li gninsulfoic acid, the dodecylsulfuric acid ester and sulfonic acids of fatty alcohol-ethylene oxide adducts. The sulfonated benzimidazole derivatives preferably contain two sulfonic acid groups and a fatty acid residue with about 8 to 22 carbon atoms. Alkylaryl sulfonates are e.g. the sodium, calcium or triethanolamine salts of dodecylbenzenesulfonic acid, dibutylnaphthalenesulfonic acid or a naphthalenesulfonic acid / formaldehyde condensation product.

Ais nonionic surfactants primarily to choose from, the polyglycol ether derivatives of aliphatic or cycloaliphatic alcohols, saturated or unsaturated fatty acids and alkylphenols, which contain 3 to 30 glycol ether groups and 8 to 20 C atoms in the (aliphatic) hydrocarbon radical and 6 to 18 C atoms in the alkyl radical can contain. Other suitable nonionic surfactants are the water-soluble, 20 to 250 ethylene glycol ether groups and 10 to 100 propylene ether groups13

5

10th

15

20th

25th

30th

35

40

45

50

55

CH 685 674 A5

holding polyethylene oxide adducts with polypropylene glycol and alkylpolypropylene glycol with 1 to 10 carbon atoms in the alkyl chain. The compounds mentioned usually contain 1 to 5 ethylene glycol units per propylene glycol unit.

Examples of non-ionic surfactants are:

Nonylphenolpolyäthoxyäthanole, castor oil polyglycol ether, Polypropylenpolyäthylenoxyd-adducts, Tri-butylphenoxypolyäthoxyäthanol, polyethylene glycol and octylphenoxypolyäthoxyäthanol. Fatty acid esters of polyoxyethylene sorbitan, such as the polyoxyethylene sorbitan trioleate, are also suitable.

The cationic surfactants are primarily quaternary ammonium salts which contain at least one alkyl radical having 8 to 22 carbon atoms as N substituents and, as further substituents, have low, optionally halogenated alkyl, benzyl or low hydroxyalkyl radicals . The salts are previously in the form of halides, methyl sulfates or ethyl sulfates, e.g. the stearyitrimethylammonium chloride or the benzyldi (2-chloroethyl) ethylammonium bromide.

On the one hand, highly preferred for the production of spontaneously dispersible concentrates according to the invention are phosphoric acid ester surfactants, such as

TristyryIphenolpolyoxyäthylen-1 8-mono / dimethylphosphorsäureester o

II

Diphasol® 3873 (CIBA-GEIGY), or the identical Sermul® EA 188 (SERVO), a mixed emulsifier, each consisting of 50% of the two compounds with the formulas:

o c9 h19 - ^} - 0 (-ch2-ch2-o VP— ° ch3

oh oh

C9 H19- ^ 3 (-CH2 * CH2 '° W-P-OCH3

och3

Diphasol® 3873 (CIBA-GEIGY)

(Surfactant 508, CIBA-GEIGY);

14

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

O

(—C h2 —-ch2 — o) 2-p och3

och3

(Surfactant 508, CIBA-GEIGY);

Tinovetin® JU (CIBA-GEIGY), a hydroxybiphenyl-10-ethoxy-phosphoric acid ester butyl-mono-4-ethoxy-phosphoric acid ester (Zerostat® AT, CIBA-GEIGY), or

O

I.

ch3- (ch2) j-çh-ch2-0 (-ch2-ch2-o) 6-p och3

och3

(Zerostat ® AN, CIBA-GEIGY)

and on the other hand betaine compounds, i.e. amphoteric, salt and water-free imidazole derivatives, such as:

where Me [+] stands for hydrogen, alkali and alkaline earth atoms and Rx for Cr to C32-alkyl or C2 to C32-alkenyl groups.

So-called "muiti-functional glucose derivatives" are also used, such as Glucate® SS (methyl glucose sesquistearate) and Glucamate® SSE-20 (PEG-20 methyl glucose sesquistearate) from Amer-chol, Edison, N.J.

Invadin® JFC 800% (CIBA-GEIGY) is an anhydrous, tert. Octylphenyipolyoxyethylene ether surfactant with 9 to 10 oxyethylene groups.

Pharmaceutically acceptable solvents which can be used as hydrotropes or as co-emulsifiers can be used, e.g .:

Esters of an aliphatic alcohol (C3 to Cis) with an aliphatic carboxylic acid (C10 to C22) such as isopropyl laurate, hexyl laurate, decyl laurate, isopropyl myristate (IPM), isopropyl palmitate and lauryl myristate; Hydrocarbons with a straight carbon chain (C | 2 to C32), which is substituted by 6 to 16 methyl groups and can have 1 to 6 double bonds, of which terpenes such as polymethylbutane and polymethylbutenes may serve as examples.

Mono-esters of ethylene glycol or propylene glycol with an aliphatic carboxylic acid (C6 to C22), such as propylene glycol monolaurate and propylene glycol monomyristate.

Esters of an aliphatic alcohol (C12 to C22) with lactic acid, e.g. Myristyl or preferably lauryl lactate; Mono-, di- or triesters of glycerol with an aliphatic carboxylic acid (C6 to C22), e.g. Glyceryl caprylate, or Miglyol® 812 neutral oil (oleum neutral).

Esters of a poly (2-7) ethylene glycol glycerol ether with at least one free hydroxyl group with an aliphatic carboxylic acid (C6 to C22), e.g. aliphatic alcohols (C12 to C22), including

ch2 ch2 oh

Rx-C

15

5

10th

15

20th

25th

30th

35

40

45

50

55

CH 685 674 A5

Dodecanol, tetradecanol, oleyl alcohol, 2-hexyldecanol and 2-octylidecanol. Esters with at least one free hydroxyl group, from poly (2-10) glycol with an aliphatic carboxylic acid (C6 to C22), monoethers from a polyethylene glycol with an aliphatic alcohol (C12 to Cis), e.g. Polyoxyethyne (Cio) octyl ether.

Heterocyclic compounds, e.g. 1-methyl-2-pyrrolideone. Biosurfactant esters of the general formula:

R4 eoo — Rfi

(XXIV)

wherein R4 citronellyl, geranyl, farnesyl, phytyl or isophytyi and R5 is a Cr to C32 alkyl, or a C2 to C32 alkenyl or alkapolyene group.

Before being added to the spontaneously dispersible concentrates, all technical surfactants were filtered or chromatographed over neutral aluminum oxide with an inert solvent such as e.g. Cleaned tetrahydrofuran, ethyl alcohol or methylene chloride.

Vitamins and provitamins (such as vitamin A, retinol, tocopherols), as well as selected penetration enhancers ("flux enhancers") and radical scavengers are suitable as additives in the spontaneously dispersible concentrates according to the invention.

The daily dose required for pharmaceutical use is 0.001 to 25 mg / kg body weight, if possible divided into 2 to 3 individual doses. For this purpose, the new esters with apocarotenols or the spontaneously dispersible concentrates with these compounds can be used in the common pharmaceutical preparations and dosage forms such as pills, tablets, capsules, powders, granules, pellets, solutions, ampoules, emulsions, creams or suppositories together with the usual carriers and / or incorporate diluents and stabilizers.

The active ingredients or active ingredient mixtures forming the subject of the invention, as well as the spontaneously dispersible concentrates which contain these active ingredients or active ingredient mixtures, can be administered to humans orally, by injection (intravenously, subcutaneously or intramuscularly) or in another way. When prepared as solid dosage forms for oral use, it can be in the form of tablets, granules, pellets, powders or capsules, etc. The preparations can contain additives, e.g. a drug carrier such as a saccharide or cellulose base, a binder such as starch paste or methyl cellulose, a filler or a disintegrant, etc. - using additives which are usually used in the manufacture of medical or paramedical formulations. When the active ingredients or active ingredient mixtures according to the invention are administered orally as liquid dosage forms, they can have any form selected from aqueous preparations for internal use, from suspensions, emulsions and syrups etc., and they can also be in the form of vacuum-dried preparations, which are brought into solution or emulsion before use.

If the active compounds or active compound mixtures according to the invention are prepared in the form of aqueous solutions, suspensions or oily or aqueous emulsions, preferably as microemulsions from the spontaneously dispersible concentrates in accordance with the invention, they can also be injected. However, the injection solutions are usually prepared shortly before use by dissolving or suspending the extracts or concentrates in aqueous, liquid media such as sterile water, glucose solution or physiological saline.

If necessary, commonly used solvents, stabilizers, preservatives and additives for the preparation of isotonic solutions can be added to an injection preparation. The injection preparations obtained in this way are administered intravenously, intramuscularly, subcutaneously or in another suitable manner.

The present invention also relates to pharmaceutical preparations which contain the active substances or active substance mixtures and / or the described, spontaneously dispersible concentrates for combating the growth of tumor cells. The pharmaceutical preparations according to the invention are those for enteral (such as oral or rectal), as well as for parenteral or topical administration to warm-blooded animals, which contain the spontaneously dispersible concentrate alone or together with a pharmaceutically usable carrier material.

The dosage of the concentrates according to the invention depends on the warm-blooded species, the age and the individual condition, and on the mode of administration. For example, to achieve a killing effect of tumor cells on warm-blooded animals with low body weight, such as Mice, rats and hamsters, doses in the range of about 0.1 to 50 mg / kg of body weight when administered successively and doses in the range of 0.05 to 5 mg / kg of body weight when administered intraperitoneally.

The oral and rectal forms of the new pharmaceutical preparations contain between 1-95%, preferably between 10-95%, in particular between 20-95% of the spontaneously dispersible concentrate according to the invention. You can e.g. Available in unit dose form, i.e. as dragées, micropellets, tablets, suppositories or ampoules and above all as capsules.

16

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

Suitable pharmaceutically usable excipients for the oral forms are mainly fillers, such as sugar (e.g. lactose, sucrose, mannitol or sorbitol), cellulose preparations and / or calcium phosphates (e.g. tricalcium or calcium hydrogen phosphate), as well as binders such as starch paste using, among others. Corn, wheat, rice or potato starch, gelatin, tragacanth, methyl cellulose, hydroxyl methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and / or polyvinylpyrrolidone and / or disintegrants (if desired), such as the above-mentioned starches, furthermore car-boxymethyl starch, cross-linked Polyvinyl pyrrolidone, agar, alginic acid or a salt thereof, such as Sodium alginate.

The flow regulating agents are e.g. the polyethylene glycols No. 200 to 600 and higher are suitable.

The gelatin capsules preferred as a dosage form in humans are provided with suitable coatings, one of the things being: Concentrated sugar solutions - which may contain arabic gum, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide - lacquer solutions (aqueous or those that have been prepared using organic solvents), or enteric coatings from solutions of suitable cellulose preparations, such as microcrystalline cellulose (Avicel® ), Acetyl cellulose phthalate, hydroxyl methyl cellulose phthalate (Metolose®), hydroxyl propyl methyl cellulose acetate succinate (AQOAT®) or a copolymer such as Eudragit® L30D.

Push-fit capsules made of gelatin and a plasticizer, such as glycerol or sorbitol, can be used as a particularly suitable pharmaceutical dosage form which can be used orally. The soft or hard gelatin capsules, as well as those made from AQOAT® (hydroxypropyl methyl cellulose acetate succinate), can be the spontaneously dispersible concentrate according to the invention in a mixture with fillers, such as lactose, binders such as starch and / or lubricants such as talc or Magnesium stearate and optionally with stabilizers and antioxidants such as contain a-, ß- or 7-tocopherol. The use of suitable liquids such as liquid polyethylene glycols No. 200 to 600 can be recommended as a diluent, but stabilizers and antioxidants can also be added.

For parenteral administration, the concentrates according to the invention are mixed with distilled water. The resulting aqueous injection microemulsion can contain viscosity-increasing substances, e.g. Na carboxymethyl cellulose, sorbitol, mannitol and / or dextran, and optionally also stabilizers and antioxidants are added.

The pharmaceutical preparations for parenteral use preferably contain between 0.1 to 60%, and mainly between 1 to 40% of the spontaneously dispersible concentrate according to the invention.

As topically applicable preparations, which are primarily suitable for the prophylaxis and therapy of skin cancers, come e.g. Creams, ointments, pastes, pomades, foams, tinctures and solutions into consideration, which contain between 0.01 to 70% of the concentrate according to the invention.

For creams and oil-in-water emulsions which contain more than 50% water, primarily fatty alcohols, e.g. Lauryl, cetyl or stearyl alcohol, liquid to solid waxes, e.g. Isopropyl myristate, wool or beeswax and / or hydrocarbons such as Vaseline (petrolatum) or paraffin oil. For the emulsification of these oily bases, surface-active, pharmaceutically compatible substances with predominantly hydrophilic properties, such as e.g. Non-ionic emulators, especially fatty acid esters of polyalcohols or ethylene oxide adducts (such as polyglycerol fatty acid esters or polyethylene sorbitan fatty acid esters) with an HLB value of less than 8. Additions to the water phase include Agents which reduce the drying out of the creams, e.g. Polyalcohols such as glycerin, sorbitol, propylene glycol and / or polyethylene glycols No. 200 to 600, also preservatives, fragrances etc.

Ointments are water-in-oil emulsions that contain up to 70%, but preferably between 20 and 50% water or aqueous phases. The fatty phase is primarily hydrocarbons, e.g. Vaseline, paraffin oil and / or hard paraffins in question, which suitable hydroxide compounds to improve water binding capacity, e.g. Fatty alcohols or esters, including cetyl alcohol or wool wax alcohols.

In some cases, emulsifiers with an HLB value of 8 to 16, e.g. Sorbitan fatty acid esters (such as sorbitan isostearol) are added. Additions to the water phase include Humectants, such as polyalcohols (glycerol, propylene glycol, sorbitol and / or polyethylene glycols No. 200, 400, 600); also preservatives, fragrances etc.

Fatty ointments are anhydrous and mainly contain hydrocarbons, e.g. Paraffin, petroleum jelly and / or liquid paraffins; also natural or partially synthetic fats such as Co-fatty acid triglyceride, further: fatty acid partial esters of glycerin, e.g. the fatty alcohols, emulsifiers and / or additives mentioned in connection with the ointments, which increase the water absorption capacity.

Pastes are creams and ointments with secretion-absorbing powder components, such as metal oxides (such as titanium oxide or zinc oxide), talc and / or aluminum silicates, which have the task of binding moisture or secretions.

Foams are administered from pressure containers and are aerosol-form oil-in-water emulsions of the spontaneously dispersible concentrates according to the invention, whereby halogenated hydrocarbons (such as chlorofluoro-lower alkanes: e.g. dichlorodifluoromethane and dichlorotetrafluoroethane) as

17th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

Propellants are used. In addition, there may be the usual additives such as preservatives, etc.

PROCESS EXAMPLES for the preparation of esters with apocarotenols of the formulas (I) to (IX): a) Preparation of apo-β-8'-carotenoid-10-undecenoate

85 mg of apo-β-8'-carotenol (synthesis see R. Ruegg et al., Helv. Chim Acta 42, 847-864) 1959), 100 mg of pyridine and 100 mg of dimethylformamide in 20 ml of chloroform are added at 0 ° C and 100 mg of 10-undecenoyl chloride in 10 ml of chloroform were added dropwise under a protective gas supply. After stirring for 1 hour at 0 ° C and stirring for 2 hours at 20 to 40 ° C, the mixture is diluted with water and shaken out. The residue is dried gently and then chromatographed on a silica gel column using hexane / ethyl acetate (90:10) as the eluent.

The apo-β-8'-carotinyl-10-undecenoate is obtained with a melting point of 76-77 ° C, after sintering from 75 ° C; UV Xmax. 425 nm (in methylene chloride); Rf value in TLC with hexane / ethyl acetate (80:20) 0.92.

IR 3054 cm-1

8 (Olefin.CH)

2855 cm-1

v (CH)

1724 cm-1

v (C = 0) ester

1639 cm-1

v (C-C)

1465 cm-i

S (CH)

1374 cm-1

8 (CHs)

1185 cm-i v (C-O) ester

1032 cm-1

v (C-O)

914 cm-1

8 (CH) from olefin

trans disubst. C = C

Azafrinyl-10-undecenoate is also prepared in an analogous manner:

20th

Refractive index n— of 1,44210; UV Xmax. 420.0 nm (in methylene chloride);

Rf value in TLC with hexane / ethyl acetate (80:20): 0.94.

b) Preparation of apo-β-8'-carotinyl-10-undecenoate

Starting material: 8'-apo-ß-carotene-8-acid ethyl and / or methyl ester. In a suitable apparatus with magnetic stirrer, N2 inlet tube, reflux condenser and septum, 2 g of 8'-apo-ß-carotene-8'-acidic methyl ester are added to 25 ml of EfeO, the mixture is cooled to -35 ° C. with N2 and stirring and added dropwise the calculated amount of DIBAH (diisobutylaluminium hydride) in hexane. When the addition is complete, the cooling bath is removed.

You let come on RT. After the reduction has been completed, very little ice is added very carefully and is continued until it becomes cloudy. Cool now. AI (OH) 3 fails slowly. After the precipitation has ended, the mixture is filtered through sufficient Celite, washed with MeOH and the filtrate is evaporated. When the residue begins to crystallize, it can be recrystallized from hot benzene and with the addition of hexane without further purification. Yield 1.2 g of brown-red crystals; UVA / IS Xmax. 425, 452 nm (in Et20)

Esterification of 8'-apo-ß-carotin-8'-ol with undecenoyl chloride.

400 mg of C3o alcohol, 20 ml of Et20 and 1 ml of pyridine are added dropwise with stirring and under N2 at -30 ° C with 0.22 ml of undecenoyl chloride. The mixture is slowly brought to RT, diluted with petroleum ether and shaken out successively with water and brine. Then drying over MgS04, evaporation and chromatography on 40 g of silica gel (Merck, 15 to 40 µm grain size) with benzene / hexane 2: 1. After discarding a yellow pre-zone, the red main zone is collected, evaporated and dried at 0.01 Torr at 40 ° C. The product crystallizes completely. Mp 76-77 ° C after sintering from 75 ° C. UV / VIS Xmax 425.5, 452.5 nm (in EfeO). CI-MS (with very gentle chemical ionization) with peaks at 575 and 401 nm; m / z. Mw. (M + +1); IR spectrum corresponds to ester peak at 1724 cm-1.1H-NMR spectrum.

18th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

c) Preparation of azafrinyl-10-undecenoate

Starting material Azafrin, isolated from rhizomes from Escobedia scrabifolia. Preparation of the methyl ester from the C27 acid with diazomethane / ether, cf. W. Eschenmoser and C. H. Eugster, Helv. Chim. Acta, SS, 1722 (1975), reduction with DIBAH analogous to method b); no shaking out, but direct filtration with the addition of 0.5% Hunig base. Evaporate the filtrate, take up the residue in Et20H and separate the water present, dry over MgSO4, filter, evaporate, chromatograph the residue with toluene / hexane on a CaC03 column 4.3 x 20 cm. The main zone is eluted with toluene / isopropyl alcohol (99: 1), the filtrate i.V. brought to dryness. After recrystallization from benzene / hexane and AcOEt / hexane, bright yellow, very acid-sensitive needles with an mp of 135 ° C. are obtained.

Formation of azafrin-10'-ol [undec-10-enoyl] ester.

Equipment as in method b). Preparation of 450 mg of the compound with formula

1.5 ml pyridine, 2 ml DMF, 20 ml Et20; N2, stirrer, septum, temperature minus 40 to minus 50 ° C, addition of 0.20 ml undecenoyl chloride. Allow to come to 0 ° C, then add another 0.20 ml undecenoyl chloride and stir at RT for 31/2 h. Dilute with Et20 and H2O, phase separation in the separating funnel, add another H20, then brine, dry over MgS04; evaporate. Dissolve the residue in a little CH2CI2 / hexane (10:15) + 0.25% Hünig base (N, N-diisopropylethylamine) and chromatograph on a Se-phadex LH-20 column 3.6 x 34.5 cm. Catch the broad, yellow-orange zone and evaporate. Second chromatography passage on a silica gel column 3.3 x 17.5 cm with toluene / isopropyl alcohol (99: 1 to 98: 2), which gives a good separation. Collect the light orange fraction, evaporate, dry 0.002 Torr at 42 ° C. The product is obtained: 376 mg as a viscous, light orange-red oil.

AZAFRINYL-10-UNDECENOAT

Characterization: UV / VIS (Et20): sharp three-band spectrum Xmax. 374, 394.5, 419 nm.

IR spectrum (CH2CI2): carbonyl band pronounced at 1723 cm-1

CI-MS: m / z 579 (M + +1 weak), 395 (M + + 1- undecenylic acid, very strong),

561 (M + + I-H2O), 593 (M + + 1-H20-H20).

COMPOSITION EXAMPLES of spontaneously dispersible CONCENTRATES according to the invention which contain the active compounds of the formulas (I) to (IX) and which, when diluted with water, give thermodynamically stable ULTRAMICROEMULSIONS with a hydrodynamic radius of 1.5 to 3.0 nm a) 0.5 to 30% by weight of one or more active compounds of the formulas (I) to (IX)

1 to 40% by weight isopropyl myristate, isopropyl palmitate or Miglyol® 812 (Dynamit Nobel)

0 to 45% by weight of a phosphoric acid ester surfactant, e.g. Diphasol® 3873 (CIBA-GEIGY), tenside 508 (CIBA-GEIGY), Zerostat® AN or AT (CIBA-GEIGY), Tinovetin® JU (CIBA-GEIGY), Soprophor® FL (Rhône-Poulenc) 5 to 90 wt. -% Invadin JFC 800% (CIBA-GEIGY).

b) 0.001 to 30% by weight of one or more antitumoral active substances of the formulas (I) to (IX) 0.001 to 50% by weight of one or more biosensitive esters of the general formula ch2oh oh azafrinol

(xxi)

Oh

19th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

CH 685 674 A5

r4 eoo — Rg

(XXIV)

where FU means Citroneilyi, Geranyl, Farnesyl, Phytyl or Isophytyl and Rs is a Cr to C32-Akyl, or C2 to C32-Alkenyl or Alkapolyengruppe,

0.001 to 90% by weight of a pharmaceutically compatible surfactant or surfactant mixture 0 to 10% by weight of a vitamin or provitamin

0 to 10% by weight of a penetration enhancer or a stabilizer and carriers and / or diluents.

c) 10% by weight of an oily antitumor active substance of the formulas (I) to (IX)

0 to 20% by weight of isopropyl myristate, isopropyl palmitate or Miglyol® 812 or one or more of the biosurfactant esters of the general formula:

r4 — eoo — r5

(XXIV)

where R4 means citronyl, geranyl, farnesyl, phytyl or isophytyl and Rs is a Cr to C32-alkyl, or C2 to C32-alkenyl or alkapolyene group,

35 to 45% by weight Invadin® JFC 800%

35 to 45% by weight of Soprophor® FL.

d) 2 to 5% by weight of a crystalline active ingredient of the formulas (I) to (IX) 10 to 20% by weight of isopropyl myristate, isopropyl palmitate or Miglyol® 812 or one or more of the biosurfactant esters of the general formula:

r4 — eoo — r6

(XXIV)

where R4 means citronyl, geranyl, farnesyl, phytyl or isophytyl and Rs is a Cr to C32-alkyl, or C2 to C32-alkenyl or alkapolyene group,

35 to 45% by weight Invadin® JFC 800%

35 to 45% by weight of Diphasol 3873 or Soprophor® FL.

EXAMPLE for the pharmaceutical production of a system preparation with concentrates according to the invention in the form of “multiple units”.

a) Granulation

Metolose® 90 SH-4000 (Shin-Etsu Chemical) 90.0 g

Avicel® PH-101 80.3 g

CONCENTRATE according to the invention 139.4 g

Aerosil® 200 80.3 g

£ 390.0 g

Granulate / form in a high-speed mixer or in a rotary bed with the addition of 110 g ethanol, break, seven 18 to 42 mesh, dry for 24 hours at 40 ° C.

b) MSR and RETARD equipment in a rotary bed with AQOAT® AS-HG (Shin-Etsu Chemical) and talc. c) Composition of finished granules / or. Micropellets

Core material 44%

CONCENTRATE according to the invention 25%

MSR coating 31%

E 100%

N.B .: MSR = gastric juice resistance. The pellets / granules according to a) can also be filled directly into capsules made from AQOAT® (HPMC-AS-M or HPMC-AS-N) 20 without filming

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

are sealed with 1: 1 acetone / ethanol and thus adequately control the functions of the MSR and the delayed release (retard).

Biological tests.

The antitumor effect of spontaneously dispersible concentrates with active ingredients according to the preparation examples a) to d) for the esters of apocarotenols and the work-up examples a) to d) for spontaneously dispersible concentrates is confirmed on the basis of the following test results:

1. In vitro tests with suitable tumor cell lines

A biological assay system has been developed that works with microtiter plates and dilution series. Prepare 104 / ml tumor cells in culture medium RPMI 1640 with 10% fetal calf serum (GIBCO); they are sown so leaky that they can grow during the assay in so-called non-confluent monolayers. The sample is added after 6 to 24 hours, with 100 (il per row, which is mixed with 100 pj medium in the 1st hole. Half of this is removed and introduced into the following hole, again mixed with 100 ml medium, and so on a geometric dilution series nVz is created.

The samples are incubated in the plaque assay for 3 to 5 days at 37 ° C with 31/2% CO2. Then dye / fix with 0.1% crystal violet (Fluka, Buchs) in a solution of 70% methanol, 1% formaldehyde, 29% water. The evaluation is carried out on a microscope, magnification 300 times. The greatest cytotoxic dilution is determined. The quantitative evaluation can also be carried out by means of scanning and absorption measurement on the spectrophotometer.

CYTOTOXICITY TEST

with Py6 cells (virus-transformed mouse fibroblasts)

Examination of a 2% concentrate of different composition with C 11: 1 AZAFRINYLESTER ACTIVE SUBSTANCE

CONCENTRATE

After 24 h cell toxic until:

After 48 h cell toxic until:

After 72 h cell toxic until:

No. 1 with IPM

1: 800,000

1: 3,200,000

1: 6,400,000

No. 2 with C 11-1-CITRON ELLYLESTER

1: 800,000

1: 6,400,000

1: 6,400,000

No. 3 with IPM and AOT

1: 800,000

1: 3,200,000

1: 6,400,000

No. 4 with C 11: 1-CITRO and CAA

1: 1,600,000

1:12 800,000

1:25 600,000

No. 1

2% by weight of active substance C 11: 1-AZAFRINYLESTER

12% by weight IPM (= isopropyl myristate)

86% by weight Invadin JFC 800% / Soprophor FL, 50:50

No. 2 with 12% by weight C 11: 1 citronellylester instead of IPM, otherwise the same composition as No. 1

No. 3 with 86% by weight Invadin JFC 800% / Soprophor FL / AOT, 40:40:20

No. 4 with 12% by weight C 11: 1 citronellylester instead of IPM and with 86% by weight Invadin JFC 800% / Soprophor FL / Amphonyl CAA 40:40:20

Microemulsions 1:10 000 (100 ppm W.S.), or 6 mg W. S. in 60 ml dist. Water N.B .: AOT (Fluka 86 139) = bis-2-ethylhexyl sulfosuccinate Na salt.

21

5

10th

15

20th

25th

30th

35

40

45

50

55

CH 685 674 A5

CYTOTOXICITY TEST

with Py6 lines (virus-transformed mouse fibroblasts)

Testing different concentrates of the same composition, but with different APOCA-ROTINYL / AZAFRINYL esters (calculated on 2% concentrate)

PREPARATION 48 h 72 h 96 h

In dilution In dilution In dilution effective to 1: effective to 1: effective to 1:

8-APOCAROTIN-10-UNDECENOAT 160,000 160,000 320,000

C 11: 1-AZAFRINYLESTER 320,000 320,000 640,000

CYTOTOXICITY TEST

with Py6 cells (polyoma virus transformed 3T3 mouse fibroblasts)

PREPARATION CELL TOXISGH to: CELL TOXIC to:

EXPOSURE 72 h EXPOSURE 72 h to concentrate calculated on active substance. calculated

C3o stearate 1

160

000

16 million

8-APOCAROTIN-10-UNDECENOAT 1

640

000

32 million

8-APOCAROTIN-LAU RAT 1

512

000

51 million

8-APOCAROTINE PALM ITAT 1

128

000

12.8 million

AZAFRINYL-10-UNDECENOAT 1

256

000

25.6 million

VITALITY TEST

with human tumor cell lines

A

HL 60 promyelolytic LEUKEMIA proliferation test (tritium: 1 (xCi / well H +) 1 x 104 cells per well

PREPARATION

10-5

IO "4

10-3

0.2 ppm W.S.

2 ppm W.S.

20 ppm W.S.

8-APOCAROTIN-10-UNDECENOAT

11.8%

3.5%

3.7%

C 11: 1 -AZAFRINYLESTER

32.4%

12.7%

2.7%

ID:% vitality after 48 hours with 2% MARIGENOL® CONCENTRATES, as an aqueous ultra-microemulsion (in dilution 1: 100,000, 1:10,000, 1: 1000) added to the medium once. When assessing, consider the relatively short exposure time of 48 hours. Controls: 76,589 cpm.

Test carried out by Dottoressa Anna Guarini, Università degli Studi di Torino, Clinica medica. 24-27.1. and February 14-17, 1994

VITALITY TEST

with human tumor cell lines

B

K 562 LEUKEMIA

Proliferation test (tritium: 1 nCi / well H +) 2 x 104 cells per well

PREPARATION

10-5

10-4

IO "3

0.2 ppm W.S.

2 ppm W.S.

20 ppm W.S.

8-APOCAROTIN-10-UNDECENOAT

28.5%

1.1%

0%

C 11: 1 AZAFRINYLESTER

68.6%

8.2%

0%

22

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

IDENTIFICATION:% vitality after 48 hours with 2% MARIGENOL® CONCENTRATES, added once to the medium as an aqueous ultra-microemulsion. When assessing, consider the relatively short exposure time of 48 hours. Controls: 247 125 cpm.

Tests carried out by Dottoressa Anna Guarini, Università degli Studi di Torino, Clinica medica.

VITALITY TEST

with human tumor cell lines

C.

TOM MELANOM

Proliferation test (tritium: 1 | iCi / well H +)

1 x 104 cells per well

2 x 104 cells per well

PREPARATION

10-5

10 "4

10-3

0.2 ppm W.S.

2 ppm W.S.

20 ppm W.S.

8-APOCAROTIN-10-UNDECENOAT

69.1%

65.5%

17.2%

C 11: 1 AZAFRINYLESTER

57.9%

56.8%

9.2%

ID:% vitality after 48 hours with 2% MARIGENOL® CONCENTRATES, as an aqueous ultramicroemulsion (in dilution 1: 100,000, 1:10,000 and 1: 1000) added to the medium once. When assessing, consider the relatively short exposure time of 48 hours. Controls: 57 816 cpm.

Tests carried out by Dottoressa Anna Guarini, Università degli Studi di Torino, Clinica medica.

VITALITY TEST

with human tumor cell lines

D

ADK: Human pulmonary adenocarcinoma proliferation test (tritium: 1 nCi / well H +) 2 x 104 cells per well

PREPARATION

10-5

10-t 10-3

0.2 ppm W.S.

2 ppm W.S. 20 ppm W.S.

8-APOCAROTIN-10-UNDECENOAT

11.7%

6.7% 0%

IDENTIFICATION:% vitality after 48 hours with 2% MARIGENOL® CONCENTRATES, added once to the medium as an aqueous ultra-microemulsion. When assessing, consider the relatively short exposure time of 48 hours. Controls: 57 816 cpm.

Tests carried out by Dottoressa Anna Guarini, Università degli Studi di Torino, Clinica medica.

VITALITY TEST

with human tumor cell lines

E

HL 60 promyeolytic LEUKEMIA proliferation test (tritium: 1 nCi / well H +) 2 x 104 cells per well cpm = 82 667

PREPARATION

(2% MARIGENOL® CONCENTRATE)

10-4

2 ppm W.S.

IO "3

20 ppm W.S.

C-SO ALCOHOL

2.3

0.99

CSO ACID

1.7

0.75

C 11: 1-β-8'-APOCAROTINYLESTER

1.7

1.5

ZEAXANTHIN-di-UNDECENOAT

3.5

1.2

b-CAROTINE

3.2

1.1

IDENTIFICATION:% vitality after 48 hours with 2-% MARIGENOL® CONCENTRATES, as aqueous Ul23

5

10th

15

20th

25th

30th

35

40

45

50

55

60

CH 685 674 A5

tramicroemulsion added to the medium once. When assessing, consider the relatively short exposure time of 48 hours. Controls: 82,667 cpm.

Tests carried out on June 7-10, 1994 by Dottoressa Anna Guarini, Università degli Studi di Torino, Clinica medica.

VITALITY TEST

with human tumor cell lines

F

K 562 Chronic LEUKEMIA (Leucemia mieloid cronica, crisi blastica) proliferation test (tritium: 1 nCi / well H +) 2 x 104 cells per well cpm = 138 833

PREPARATION

(2% MARIGENOL® CONCENTRATE)

10 "4

2 ppm W.S.

IO "3

20 ppm W.S.

C-30 ALCOHOL

37.1

0.7

C 30 ACID

18.6

0.5

C 11: 1-β-8'-APOCAROTINYLESTER

17.4

0.4

ZEAXANTHIN-di-UNDECENOAT

38.6

1.0

b-CAROTINE

18.5

0.6

IDENTIFICATION:% vitality after 48 hours with 2% MARIGENOL® CONCENTRATES, added once to the medium as an aqueous ultra-microemulsion. When assessing, consider the relatively short exposure time of 48 hours. Controls: 138 833 cpm.

Tests carried out on June 7-10, 1994 by Dottoressa Anna Guarini, Università degli Studi di Torino, Clinica medica.

LONG-TERM TRIAL

In order to find out whether significant changes to the tumor cell were to be found after prolonged exposure - be it in the direction of regression or redifferentiation - a long-term experiment was also carried out with Py6 cells, which was carried out over 41 days. (Experiment from May 24, 1994 to June 29, 1994). Non-transformed 3T3 mouse fibroblasts were used as controls, as were the poly-virus-transformed 3T3 MT 18 cells. The following were tested as lead preparations: a 2% PHYSALIEN concentrate (zeaxanthin-di-palmitate) and a 2% c11: 1-ß-8'-APOCAROT! NYLESTER concentrate.

Details of the test protocol: After sowing, the line growth is initially slow. After 12 days, dose-dependent, many necrotic cells appear. Washing out the cultures with EDTA. New addition of medium and active ingredient-containing microemulsion in 3 different concentrations (as at the beginning of the experiment) after 13 and 20 days. Resumption of cell growth in all surviving groups. Wash out again after 34 days and add fresh medium and concentrate. The experiment was stopped after 41 days.

Result: The morphological differences between the transformed and the non-transformed cells can still be determined in the in-vitro test even at the end of the test on the surviving populations. The transformed cells have longer filopodia and retain this characteristic.

Analytical evidence

1) Identification of the APOCAROTIN-ESTER

Capillary zone electrophoresis with a device from Beckman Instruments or from BIORAD. Conditions:

DL-a-tocopherol 50 mM; Buffer pH = 9.5 Na-tetra-Borat Run 15 kV, measurement at 195 nm The active substance peak appears after approx. 4 minutes.

Resolution up to 0.5 ppm.

2) Identification of the concentrate micelles in the aqueous microemulsion / or. in the cell plasma of the tumor cells.

Same methodology as 1).

The characteristic peak for the apocarotenes ester appears after about 8 minutes; the delay compared to the pure W.S. measurement is due to the coating of the inner phase of the concentrates with surfactants. It gives an important indication of the diffusion behavior of the ultramicroemulsion.

3) Detection of membrane penetration on the tumor cell

24th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

On the light microscope (as well as on the electron microscope) it can be shown that a ring of vacuoles forms around the cell nucleus a few hours after the incubation (example 3T3 Py6 cells of the mouse; thinly sown, medium dilution of the active ingredient concentrates).

The analytical proof that these vacuoles contain the apocarotenes active substances can be provided very clearly by removing the cell plasma from the cleaned incubated tumor cells, centrifuging them and using a 0.05% solution of Uvitex® CF conc. (CIBA-GEIGY) in Ace-tonAA / asser (85:15) or by Uvitex® EBF (CIBA-GEIGY) or by Tinopal®19 GS (CIBA-GEIGY).

The apocarotenes esters used in accordance with the invention extinguish the fluorescence caused by the marker Uvitex CF conc. Or Uvitex EBF or Tinopal GS in the long-wave UV light range; a blue color develops on the thin-layer plate. UV scanning at 366 nm. Control using micellar capillary zone electrophoresis, (Beckman Instruments, P / ACE System 2100).

EFFECT ON BLOOD

(Compatibility of MARIGENOL® PREPARATIONS)

TOXICITY of MARIGENOL® CONCENTRATES on the BALB / c mouse

% Of blood cells

preparation

L

M

N

E

B

G 17

10-7

70 ± 6

11 ± 3

13 ± 4

6 ± 4

0

10-5

77 + 6

6 + 3

11 + 4

5 ± 4

1 + 1

10-3

69 ± 10

7 + 5

22 ± 8

2 + 2

0

G 41

10-7

77 ± 6

6 ± 3

13 ± 5

3 ± 3

0

10-s

78 ± 4

10 ± 2

10 ± 4

1 + 1

1 ± 1

10-3

80 ± 6

8 ± 2

10 + 6

12 + 1

0

G 44

10-7

74 ± 17

10 ± 1

20 ± 9

1 + 1

0

10-5

74 ± 6

9 ± 4

14 + 7

4 + 3

0

10-3

76 ± 5

6 ± 4

16 ± 8

2 ± 1

0

G 55

10-7

78 + 4

10 ± 4

10 ± 4

2 ± 1

0

10-5

69 ± 10

11 ± 3

18 ± 4

1 ± 1

0

10-3

77 ± 5

6 ± 4

14 ± 2

2 ± 1

1 ± 1

CHECKS

76 ± 5

8 + 2

15 + 4

1 ± 1

0

(Physical buffer)

G 17 2% concentrate with C 5: 0-iso-CHOLESTERYLESTER (cholesteryl-iso-valerate)

G 41 2% concentrate with C 11: 1 ERGOSTERYLESTER (ergosteryl 1O undecenoate)

G 44 2% concentrate with C 18: 2-CHOLECALCIFERYLESTER (C 18: 2-D3; vitamin Ds-linolate) G 55 2% concentrate with C 4: 1-CHOLECALCIFERYLESTER (C 4: 1-D3; vitamin D3 crotonate) Dilutions: 10 ~ 7 = 0.1 ppm concentrate; 0.002 ppm active substance 10-5 = 10 ppm concentrate; 0.200 ppm active substance

10 ~ 3 = 1000 ppm concentrate; 20,000 ppm active substance (calculated on the aqueous microemulsion)

Legend:

L = lymphocytes M = monocytes (macrophages)

N = neutrophilic granulocytes E = eosinophilic granulocytes B = basophilic granulocytes

Execution of the samples: Prof. Dott. Guido FORNI, Dotta Stefania VAI, Università di Torino, Dipartimento di Scienze Cliniche e Biologiche, Ospedale San Luigi Gonzaga, 1-10 043 ORBASSANO (TO), August / September 1993.

Test with normal 8-week female BALB / c nAncr (H-2d) mice supplied by Charles River Laboratories, Calco (Italy). IV injection twice daily for 4 weeks. of 0.250 ml aqueous microemulsion, formed from the specified concentrates, or with physiological buffer for the controls. Coloring with May Grünwald-Giemsa.

Treatment time: 28 days

Blood analysis after the last injection Number of animals: 13 groups of 5 per animal

RESULT: There are no significant differences from the controls. There could be no toxic

25th

5

10th

15

20th

25th

30th

35

40

45

50

55

60

CH 685 674 A5

of the concentrates on the leukocyte population. The erythrocyte population also showed normal values. All animals were and remained healthy until the end of the experiments.

A) Measurement data for the determination of APOCAROTINYL ESTERS 1.0 undec-10-enoic acid [8'-apo-ß-carotin-8'-yl] esters (APOCAROTIN YL-1 O-UNDECENOAT)

Mp 76-77 ° C, after sintering from 75 ° C; UVA / IS (EfcO), qual.): Xmax: sh 413, 425.5, 450.5 nm; IR (CH2CI2, selection of bands); free of OH bands, 1724 cm-1 (vs., ester carbonyl); CI-MS (NH3): 585 [M + 1, (14)] 401 [M + 1 undecylenic acid (100)], 309, [M + 1-toluene, (88)].

2.0 SCHEME of the esters below

3.0 APOCAROTIN LAURIC ACID ESTER, n = 9

Orange crystals from t-butyl methyl ester / pentane. Mp 80-81 ° C. UVA / IS (CH2CI2, quantitative): Xmax: sh 415 (72,700), 434 (103,400), 458 (92,200) nm; IR (CHCI3, selection of bands); no OH bands, 1726 cm-1 (vs., ester carbonyl); 1H NMR (CDCI3), 300 MHz: 0.888 (t, w-CHs); 1.040 [CH3 (16.17)]; 1,269 [(CH2) 9]; 1.726 [CH3 (18)]; 1.848 [CHs (19 ')]; 1.957 [CH3 (20 ')]; 1,982 [CH3 (19.20)]; 2,346 [t, a-CH2); 4.567 [s, CH2 (8 ')]; 6.1 to 6.7 ppm (vinyl protons).

4.0 APOCAROTINE PALMITAT, n = 13

Orange crystals of t-butyl methyl ester / pentane at -20 ° C. Mp 73.5-75.5 ° C, after sintering from 70 ° C; Resolidification and final mp 87-91 ° C. UV / VIS (CH2CI2, quantitative): Xmax sh 410 nm (44,000), 432 nm (59,400) 457 nm (50,800). IR (CHCI3, selection of bands); free of OH bands, 1728 cm_i (vs., ester carbonyl); 1H NMR (CDCI3), 300 MHz: 0.888 (t, w-CH3); 1.038 [CH3 (16.17)]; 1.264 [(CH2) 16]; 1.725 [CH3 (18)]; 1,846 [CH3 (19 ')]; 1.956 [CH3 (19.20)]; 2,344 [t, a-CH2); 4.565 [s, CH2 (8 ')]; 6.1 to 6.7 ppm (vinyl protons).

5.0 APOCAROTINE STEARATE, n = 15

Orange crystals from t-butyl methyl ester / pentane at - 20 ° C. Mp 80-81 ° C, after re-solidification 80-89 ° C; IR (CHCI3, selection of bands); free of OH bands, 1728 cm_i (vs., ester carbonyl); UV / VIS (CHCI2), quantitative): Xmax sh 410 nm (61 700), 433 nm (87 900) 458 nm (76 400). 1H-NMR (CDCI3),

300 MHz: 0.888 (t, co-CH3); 1.039 [CH3 (16.17)]; 1.264 [(CH2) 16]; 1.726 [CH3 (18)]; 1.848 [CH3 (19 ')]; 1.957 [CHs (20 ')]; 1.982 [CH3 (19.20)] 2.345 [t, cu-CH2); 4.566 [s, CH2 (8 ']; 6.1 to 6.7 ppm (vinyl protons).

B. DATA on AZAFRINOL and AZAFRINYL-ESTERN 1.0 Properties of Azafrinol:

Mp 133-134 ° C (vacuum tube). Combustion analysis for C27H40O3 (412.6): calc. C 78.59 H 9.77%; found C 78.30 H 9.56%. UVA / IS (EtOH, log e): 432.4 (4.514), 375.2 (4.805), 396.0 (5.011), 420.0 (5.005);

[<x] - = 85.1 ° (c = 21.80 mg in 2.00 ml EtOH):

D

CD ((EtOH.c = 1,192.10-s M, Ae, major bands): 237.2 (-2.5), 370.4 (-1.4), 386.0 (-1.2), 420.4 (-1.6). IR (KBr): very broad OH bands around 3450 cm-1, no bands in the carbonyl region; IR (CH2CI2): 3600 cm-1, no cabonyl bands; CI-MS (NH3): 413 [ M + 1 (13)], 396 [M + 2-H20 (30)], 395 [M + 1-18 (100)].

26

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

CH 685 674 A5

2.0 Properties of AZAFRINYL ESTERS

2.1 AZAFRINYL-10-UNDECENOAT

UVA / IS (Et20, qual.): Sharp three-band spectrum with Xmax 375, 394.5, 419 nm; (Qual. Benzene): 377, 402, 428 nm. IR (CH2CI2, selection of bands): 2.78 n, (= OH-free), approx. 2.9 n = OH intermolecularly bound), 5.8 p (1723 cm-1), vs. Ester carbonyl; 1H-NMR (CDCI3), 300 MHz. Selection of bands): for the azafrinol part: recent NMR spectra of azafrin and derivatives, cf. P. Übelhart and C. H. Eugster, Helv. Chim. Acta 1982, SS, 353: 0.85, (CH3 (16)); 1.15 (CH3 (17)); 1.19 (CH3 (18)); 1.91 (CH3 (20 ')); 1.98 (CH3 (19.20)); for the acid part: 1.30 ((CH2) 7); 2.33 (t, a-CH2); 4.9 (m, w) -CH2); 5.8 (m, irCH).

2.2 Other esters:

öh n = 9; n = 13; n = 15, or

Azafrinyl laurate UV / VIS 376, 402, 428 nm,

Azafrinyl palmitate UV / VIS 372, 402, 428 nm

Azafrinyl stearate UV / VIS 379, 402, 428 nm.

Claims (11)

Claims 1. Spontaneously dispersible concentrates, which are diluted with water, give thermodynamically stable ultramicroemulsions with micelles that have a hydrodynamic radius of 1.5 to 3 nm, characterized in that the following components are combined to form a system: 0.001 to 30% by weight individual esters of apocarotenols of the formulas (I) to (IX), or a combination of such esters 27 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 —R. (IV) 28 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 O II CH ^ -O— C— ^ ch, £ h '4 ch., ch (VU) o ch2-o-c - R. (Vili) cho (IX) where in the formulas (I) to (IX) R 1 is a Cr to C32 alkyl, a C2 to C32 alkenyl or alkapolyen group, a C2 to C32 alkynyl group or a radical of the formulas (X), or (XI) represents: 29 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 ? H, r2 - (- ch = ch-c = ch-) n (X) Ro ÇH: CH & CH — C — ch -) - (- chzch — chz <jî -) - chzch ch, (xi) where n denotes the numbers 1, 2, 3, 4 or 5 and Ffe for one of the radicals of the formulas ch ch, ch, ch3 * 0ch ' çh, ^ Jy ° ch3 ch, ch, ch. ch CJJjpH, Ä CH'-kAoH ch, ch ch, oh oco ch .ch, ch, Oh 30th 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 OCO CH, or —CO stands, 0 to 40% by weight of a pharmaceutically acceptable solvent or solvent mixture serving as a hydrotrope or co-emulsifier 0.001 to 90% by weight of a pharmaceutically acceptable surfactant or surfactant mixture, 0 to 10% by weight of a vitamin or provitamin, 0 to 10 wt .-% of a stabilizer or a penetration enhancer and carriers and / or diluents. 2. Spontaneously dispersible concentrates according to claim 1, characterized in that the following components are combined to form a system: 0.5 to 30% by weight of one or more active compounds of the formulas (I) to (IX), 1 to 40% by weight of isopropyl myristate, isopropyl palmitate or neutral oil (oleum neutral), 0 to 45% by weight of a phosphoric acid ester surfactant and 5 to 90 wt .-% of anhydrous tert. Octylphenylpolyoxyethylene ether surfactant with 9 to 10 oxyethylene groups. 3. Spontaneously dispersible concentrates according to claim 1, characterized in that the following components are combined to form a system: 0.5 to 30% by weight of one or more active compounds of the formulas (I) to (IX) 1 to 40% by weight of one or more biosensitive esters of the general formula (XXIV): • COO — R « (XXIV) where FU means Citroneilyi, Geranyl, Farnesyl, Phytyl or Isophytyl and Rs is a Cr to C32-Akyl, or C2 to C32-Alkenyl or Alkapolyengruppe, 5 to 90% by weight of a pharmaceutically acceptable surfactant or surfactant mixture, 0 to 10% by weight of a vitamin or provitamin, 0 to 10 wt .-% of a stabilizer or a penetration enhancer and carriers and / or diluents. 4. Spontaneously dispersible concentrates according to claim 1, characterized in that the following components are combined to form a system: 10% by weight of an oily active ingredient of the formulas (I) to (IX), 0 to 20% by weight of isopropyl myristate, isopropyl palmitate, neutral oil or one or more biosensitive esters of the general formula (XXIV): r4 — eoo r6 (xxiv) where R4 means citronyl, geranyl, farnesyl, phytyl or isophytyl and Rs is a Cr to C32-alkyl, or C2 to C32-alkenyl or alkapolyene group, 35 to 45 wt .-% of anhydrous tert. Octylphenyl polyoxyethylene ether surfactant with 9 to 10 oxyethylene groups, 35 to 45% by weight of an alkylphenol polyglycol ether phosphate surfactant or of the tristyrylphenol polyoxyethylene-18-mono / dimethyl phosphoric acid ester surfactant. 5. Spontaneously dispersible concentrates according to claim 1, characterized in that the following components are combined to form a system: 2 to 5% by weight of a crystalline active ingredient of the formulas (I) to (IX), 10 to 20% by weight of isopropyl myristate, isopropyl palmitate, neutral oil or one or more biosensitive esters of the general formula (XXIV): ra — eoo — r6 (xxi v) where R4 means citronyl, geranyl, farnesyl, phytyl or isophytyl and Rs is a Cr to C32-alkyl, or C2 to C32-alkenyl or alkapolyene group, 31 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 35 to 45 wt .-% of anhydrous tert. Octylphenylpolyoxyethylene ether surfactant with 9 to 10 oxyethylene groups, 35 to 45% by weight of an alkylphenol polyglycol ether phosphate surfactant or of the tristyrylphenol polyoxyethylene-18-mono / dimethyl phosphoric acid ester surfactant. 6. Esters of the apocarotenols of the formulas (I) to (IX) ch2 — o — c — r1 —R, CH (II) ch, O > Il ) —C — R, (iii) CH2 — O— c - r1 (iv) ch ch2 — o— c— r, (V) 32 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 fi CHr-O-C-R1 CH, CH3 CH, CH (VI) il C H2 —- O — C— (VU) fi CH2-O-c-R1 (IX) wherein Ri for a C4 to Cis-alkyi, a C2 to Cis-aikenyl, a C2 to Cis-aikapoiyen, a C2 to Ci8-alkynyl group or for a radical of the formula (XII) (XII) stands. 7. The 8'-apo-ß-carotinyl-10-undecenoate, 8'-apo-ß-carotinyl laurate, 8'-apo-ß-carotinyl palmitate, 8'-apo-ß-carotinyl stearate, and the azafrinyl-10-undecenoate , Azafrinyl laurate, azafrinyl palmitate and the azafrinyl stearate according to claim 6. 8. A process for the preparation of esters of apocarotenols according to claim 6, characterized in that an apocarotene of the formulas (XV) to (XXXIII): 33 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 h2oh (XV) (xv!) (xvii) (XVIII) (XIX) 34 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 CH 685 674 A5 îh, ÌH, : h2oh ch5 ch, ch, ch, ch, hh with the chloride or bromide of an acid of the formula (XIV): R3 COOH (xx) ch2oh (XXI) ch2oh (XXIII) (xiv) where R3 represents a C4 to Gra alkyl, a C2 to Cis alkenyl, a C2 to Cis alkapolyen, a C2 to Ci8 alkynyl group or a radical of the formula (XII): ch (xii) at a temperature of 0 to 60 ° C with inert gas supply in an inert solvent and in the presence of a catalyst. 9. A process for the preparation of 8'-apo-ß-carotinyl esters according to formula (III) according to claim 1, characterized in that the 8'-apo-ß-carotene-8'-acidic acid ester at minus 30 ° C with DIBAH ( Diisobutylaluminiumhydrid) reduced to obtain the C3o alcohol as an intermediate, which then with the chloride of an acid of the formula: 35 5 10th 15 20th 25th 30th 35 40 45 50 55 CH 685 674 A5 R, COOH is esterified in EfeO and pyridine at minus 30 ° C and then at room temperature, where Ri has the same meaning as in claim 1. 10. A process for the preparation of azafrinyl esters according to formula (VII) according to claim 1, characterized in that azafrinol is first prepared from azafrin methyl ester by means of a DIBAH reaction and this compound is then immediately mixed with the chloride of an acid of the formula: R1 COOH esterified in EfeO and pyridine at minus 30 ° C and then at room temperature, where Ri has the same meaning as in claim 1. 11. Spontaneously dispersible concentrate, containing esters of the apocarotenols of the formulas (I) to (IX) according to claim 1, as an agent for producing a pharmaceutical preparation effective against tumors, psoriasis and eczema. 36