Formulations Containing Alkylphosphocholines Using Novel Negative Charge Carriers

Abstract

The invention relates to novel medicament formulations containing, as active ingredients, alkylphosphocholines and the like, alkyl-alkanediol-phosphocholines and the like, and (ether)lysolecithines and the like, in different forms of embodiment. Said active ingredients are integral constituents of liposomes, also containing cholesterol and the like and a negative charge carrier. The medicament formulations are especially suitable for the treatment and/or prophylaxis of cancer, protozoan diseases such as leishmaniasis and amoebic diseases, acariasis and diseases caused by arthropods, and bacterial diseases, such as ehrlichiosis. Ocular diseases accompanied by uncontrolled cellular processes can also be advantageously influenced.

Description

EXAMPLES

Example Group 1

Variation of the Oleylphosphocholine/Cholesterol Ratio

Oleylphosphocholine (MG 433.61)(Ol-PC)
Cholesterol         (MG 386.66)(Chol)
Oleic acid          (MG 282.47)
NaOH                (MG 40.00)

Example 1 (a)

O1-PC, 40 mM; Chol 35mM

Ol-PC	40.0 mM; 1.73 g
Chol	35.0 mM; 1.35 g	weighed-in
		portion: 100 g = 100 ml
Oleic acid	10.0 mM; 0.283 g
NaOH	9.5 mM; 9.50 g 0.1 N NaOH

Preparation

The weighed-in portion was dissolved in 30 ml CH₂Cl₂, CH₂Cl₂ was removed, dried to a constant weight, mixed with 50 g 0.3 M 1,2-propanediol and 9.5 g 0.1 N NaOH and sufficient 0.3 M 1,2-propanediol was added to make the overall weight 100 g.

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

0.2 μ of the mixture was filtered when still warm and stored at from +4° C. to +8° C.

Observations—stable, at least 1 year at from +4° C. to +8° C.

Example 1 (b)

O1-PC, 40 mM; Chol 40 mM

0l-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in
		portion: 100 g = 100 ml
Oleic acid	10.0 mM; 0.283 g
NaOH	9.5 mM; 9.50 g 0.1 N NaOH

Preparation—as in Example 1 (a)

Observations—stable, at least 1 year at from +4° C. to +8° C.

Example 1 (c)

O1-PC, 40 mM: Chol 45 mM

Ol-PC	40.0 mM; 1.73 g
Chol	45.0 mM; 1.74 g	weighed-in
		portion: 100 g = 100 ml
Oleic acid	10.0 mM; 0.283 g
NaOH	9.5 mM; 9.50 g 0.1 N NaOH

Preparation—as in Example 1 (a)

Observations—stable, at least 1 year at from +4° C. to +8° C.

According to the results obtained in Examples 1 (a) to 1 (c), a preparation for the subcutaneous treatment of dogs can have the following composition:

	mM
	Ol-PC	(active ingredient)	35-45
	Chol	(auxiliary)	35-45
	Oleic acid	(95%)	8-12

Example Group 2

Variation of the Oleic Acid Content

The composition was varied with regard to the oleic acid content, although in all cases at a degree of protonation of 95%. These variations were examined on

Ol-PC 40.0 mM
Chol  40.0 mM

Oleic acid as sodium salt was varied between 2.0 mM and 25 mM. Some easy-to-use storable formulations are described precisely under 2 (a) to 2 (d).

Oleylphosphocholine (MG 433.61),
Cholesterol         (MG 386.66)
Oleic acid          (MG 282.47)
NaOH                (MG 40.00)

Example 2 (a)

O1-PC, 40 mM; Chol 40 mM; oleic acid 16 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in
		portion: 100 g = 100 ml
Oleic acid	16.0 mM; 0.452 g
NaOH	15.2 mM; 7.60 g 0.1 N NaOH

Preparation—as in Example 1 (a)

Observations—stable, at least for 1 year at from +4° C. to +8° C.; does not irritate tissue in dogs.

Example 2 (c)

O1=PC, 40 nM; Chol 40 mM; oleic acid 8 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in portion
		100 g = 100 ml
Oleic acid	8.0 mM; 0.225 g
NaOH	7.6 mM; 7.60 g 0.1 N NaOH

Preparation—as in Example 1 (a)

Observations—stable, for at least 1 year at from +4° C. to +8° C.; does not irritate tissue in dogs.

Example 2 (c)

O1-PC, 40 mM; Chol 40 mM; oleic acid 6 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in portion
		100 g = 100 ml
Oleic acid	6.0 mM; 0.170 g
NaOH	5.6 mM; 5.60 g 0.1 N NaOH

Preparation—as in Example 1 (a)

Observations—stable, at least for 1 year at from +4° C. to +8° C.; does not irritate tissue in dogs.

Example 2 (d)

O1-PC, 40 mM; Chol 40 mM; oleic acid 4 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in
		portion: 100 g = 100 ml
Oleic acid	4.0 mM; 0.112 g
NaOH	3.8 mM; 3.80 g 0.1 N NaOH

Preparation—as in Example 1 (a)

Observations—stable, at least for 1 year at from +4° C. to +8° C.; does not irritate tissue in dogs.

Example Group 3

Preparation of O1-PC Dispersions for Application in Tests on Animals

Oleylphosphocholines (MG 433.61) 39.20 mM
Cholesterol (MG 386.66)          41.40 mM
Oleic acid (MG 282.47)           5.66 mM
NaOH (MG 40.00)                  5.40 mM

Weighed-in portion

	Per 1 kg	Per 0.5 kg	Per 0.1 kg
	Ol-PC	17.00 g	8.5 g	1.700 g
	Chol	16.00 g	8.0 g	1.600 g
	Oleic acid	1.60 g	0.8 g	0.160 g
	NaOH, 0.1 N	54.00 g	27.0 g	5.400 g

Example 3 (a)

Preparation of a Sample Dispersion of 100 g O1-PC

The amounts of substance required for 0.11=100 ml were weighed out into a 500 ml round-bottom flask and completely dissolved in 40 ml CH₂Cl₂. The solution had to be clear and free from particles. CH₂Cl₂ was removed on a rotary evaporator under a slight vacuum and the residue was dried under vacuum to a constant weight, most simply overnight. Result: 3.44 to 3.46 g corresponding to >99% of the weighed-in portion.

The residue was mixed with a solution consisting of 90 ml 0.289 M 1,2-propanediol and 5.4 ml 0. 1 M NaOH and the overall weight was brought to 100 g. The error in volume occurring in this case with 0.289 M 1,2-propanediol was low, as the density of the dispersion corresponded to approximately 1. The mixture was heated to 55° C. with rotation and the system kept at this temperature:

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

The mixture was sterile-filtered while still warm through 0.2 μ and stored at from +4° C. to +8° C.

Observations—the dispersion is stable, storage at least 1 year. It meets all important criteria.

• Stability in storage
• Filtration under sterile conditions possible
• Heat sterilisation possible
• Synthesis for auxiliary dispensed with
• Auxiliary pharmaceutically known
• Auxiliary also has a buffer capacity at pH 5.0

Example 3 (b)

Preparation of a Dispersion of 500 g O1-PC

The amounts of substance required for 0.5 1=500 ml were weighed out in a 11 round-bottom flask and completely dissolved in 100 ml CH₂Cl₂. The solution has to be clear and free from particles. CH₂Cl₂ was removed on a rotary evaporator under a slight vacuum and the residue was dried to a constant weight, most simply under vacuum overnight. Result: 17.2 to 17.3 g, corresponding to >99% of the weighed-in portion.

The residue was mixed with a solution consisting of 450 ml 0.289 M 1,2-propanedioland 27 ml 0. 1 N NaOH and the overall weight brought to 500 g. The specific weight of the dispersion was almost 1, so the error in volume was slight. The mixture was heated to 55° C. with rotation and the system kept at this temperature using a water bath:

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

The mixture was sterile-filtered while still warm through 0.2 μ and stored at from +4° C. to +8° C.

Observations—the dispersion is stable, storage at least 1 year. It meets all important criteria.

• Stability in storage
• Filtration under sterile conditions possible
• Heat sterilisation possible
• Synthesis for auxiliary dispensed with
• Auxiliary pharmaceutically known
• Auxiliary also has a buffer capacity at pH 5.0

Example Group 4

Fatty Acid Amides of Amino Acids as Negative Charge Carriers

Oleylphosphocholine         (MG 433.61)
Cholesterol                 (MG 386.66)
N-oleoyl-alanine, Na(+)salt (MG 375.53)

Example 4 (a)

O1-PC, 40 mM; Chol 40 mM; N-oleoyl-alanine, 5 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in portion: 100 g = 100 ml
N-oleoyl-Al	5.0 mM; 0.19 g

Preparation

The weighed-in portion was dissolved in 30 ml CH₂Cl₂, the solvent was removed and the residue dried to a constant weight. The residue was brought to a total of 100 g with 0.3 M 1,2-propanediol and heated to 55° C.:

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

The mixture was filtered under sterile conditions while still warm through 0.2 μ and stored at from +4° C. to +8° C.

Example 4 (b)

O1-PC, 40 mM; Chol 40 mM; N-oleoyl-alanine, 10 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in portion: 100 g = 100 ml
N-oleoyl-Al	10.0 mM; 0.38 g

Preparation

The weighed-in portion was dissolved in 30 ml CH₂Cl₂, the solvent was removed and the residue dried to a constant weight. The residue was brought to a total of 100 g with 0.3 M 1,2-propanediol and heated to 55° C.:

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

The mixture was filtered under sterile conditions while still warm through 0.2 μ and stored at from +4° C. to +8° C.

Example Group 5

Fatty Acid Amides of Glycero-Phospho-Ethanolamine as Negative Charge Carriers

Oleylphosphocholine              (MG 433.61)
Cholesterol                      (MG 386.66)
N-oleoyl-glycero-phospho-ethanolamide, Na(+)salt (MG 501.57)

Example 5(a)

O1-PC, 40 mM; Chol 40 mM; N-oleoyl-GPE, 5 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in portion: 100 g = 100 ml
N-oleoyl-GPE	5.0 mM; 0.25 g

Preparation

The weighed-in portion was dissolved in 30 ml CH₂Cl₂, the solvent was removed and the residue dried to a constant weight. The residue was brought to a total of 100 g with 0.3 M 1,2-propanediol and heated to 55° C.:

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

The mixture was filtered under sterile conditions while still warm through 0.2 μ and stored at from +4° C. to +8° C.

Example 5 (b)

O1-PC, 40 mM; Chol 40 mM; N-oleoyl-GPE, 10 mM

Ol-PC	40.0 mM; 1.73 g
Chol	40.0 mM; 1.55 g	weighed-in portion: 100 g = 100 ml
N-oleoyl-GPE	10.0 mM; 0.50 g

Preparation

The weighed-in portion was dissolved in 30 ml CH₂Cl₂, the solvent was removed and the residue dried to a constant weight. The residue was brought to a total of 100 g with 0.3 M 1,2-propanediol and heated to 55° C.:

• a) annealing at 55° C. for 10 min
• b) ultrasound (100%) at 55° C. for 20 min

The mixture was filtered under sterile conditions while still warm through 0.2 μ and stored at from +4° C. to +8° C.

Claims

1. Pharmaceutical preparation containing as an active ingredient a) a phospholipid compound of general formula I:

2. Pharmaceutical composition according to claim 1, characterised in that the composition is present in the form of liposomes.

3. Pharmaceutical preparation according to claim 1, characterised in that it contains from 10 to 60 mol % of the active ingredient a), 10 to 65 mol % of compound b) and 3 to 30 mol % of the negative charge carrier c).

4. Pharmaceutical preparation according to claim 1, characterised in that R1, R11, R21, R31 and/or R41 is a C15-C24 alkyl radical, a C15-C24 alkenyl radical, a C15-C24 alkadienyl radical or a C15-C24 alkatrienyl radical.

5. Pharmaceutical preparation according to claim 1, characterised in that n=2.

6. Pharmaceutical formulation according to claim 1, characterised in that R2, R3 and R4, each time it occurs, is a methyl radical.

7. Pharmaceutical preparation according to claim 1, characterised in that it contains as an active ingredient 1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine (ET18OCH3).

8. Pharmaceutical preparation according to claim 1, characterised in that R1, R11, R21, R31 or R41 is oleyl.

9. Pharmaceutical preparation according to claim 1, characterised in that the negative charge carrier is selected from oleic acid, linoleic acid, lauric acid and/or palmitic acid.

10. Pharmaceutical formulation according to claim 1, characterised in that it further contains a pharmacologically acceptable excipient and/or diluent.

11. Pharmaceutical preparation according to claim 1, characterised in that components a), b) and c) together form 100 mol %.

12. Pharmaceutical preparation according to claim 1, characterised in that it contains the compound of formula I in an amount of 0.1 to 200 μmol/g.

13. Pharmaceutical preparation according to claim 1, characterised in that it is in a form suitable for intravenous, oral or subcutaneous administration and, in the case of oral administration, is formulated as a tablet or capsule.

14. Process for preparing a pharmaceutical preparation according to claim 1, characterised in that a) a compound a) is mixed as an active ingredient with a compound b) and a compound c).

15. Use of a composition according to claim 1 for preparing a pharmaceutical composition for the stimulation of leukopoesis and/or for the treatment and/or prophylaxis of acarinosis and of diseases caused by arthropods or for the treatment and/or prophylaxis of tumour diseases or for the treatment and/or prophylaxis of protozoan diseases, in particular of leishmaniasis and/or amoebic diseases, or for the treatment and/or prophylaxis of diseases caused by ascarids, in particular acarines or ticks, especially of scabies, or for the treatment and/or prophylaxis of bacterial diseases, in particular of ehrlichiosis, or for the treatment and/or prophylaxis of ocular diseases associated with cellular proliferations, in particular of detachment of the retina.

16. Use of a composition according to claim 1 for preparing a pharmaceutical preparation for veterinary medicine.

17. Use according to claim 16 for the treatment and/or prophylaxis of tumour and/or protozoan diseases.

18. Pharmaceutical preparation according to claim 1, in particular for use in veterinary medicine, characterised in that it contains cholesterol, 7β-hydroxycholesterol and/or a β-sitosterol and as a negative charge carrier c) oleic acid.

19. Use of erufosine for preparing a composition for the treatment of tumours.

20. Use according to claim 19 in combination with radiation therapy.

21. Compound of formula VII

22. Process for the preparation of alkylphosphocholine compounds as defined in claim 1, characterised in that vegetable oils, in particular olive oil, are used as a starting material.

23. Pharmaceutical preparation according to claim 1, characterised in that it contains liposomal alkylphosphocholines and is provided for oral administration.