Stereospecific lipids for locoregional therapy with long-term circulating stimuli-sensitive nanocarrier systems

Abstract

The invention relates to stereospecific lipids for the locoregional therapy with long-term circulating stimuli-sensitive nanocarrier systems. A preferred embodiment thereof is a thermosensitive liposome for treating tumors, especially urinary bladder tumors and other localized tumors.

Claims

1. A stimuli-sensitive nanocarrier system comprising at least one phosphatidyloligoglycerol of formula (IIa) ##STR00033## wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms and m is an integer from 0 to 50, wherein the linkage from the glyceride to the phosphate group is stereospecific and is in the form of an sn-3 linkage and the linkage from the phosphate group to the oligoglycerol is stereospecific and is in the form of an sn-1 linkage.

2. The stimuli-sensitive nanocarrier system of claim 1, wherein the system is a thermosensitive liposome.

3. The stimuli-sensitive nanocarrier system of claim 1, the system comprising: (i) at least one phosphatidylcholine having a main transition temperature of from 0 C. to 80 C.; and, (ii) at least one phosphatidyloligoglycerol and/or at least one phosphatidylglyceroglycol and/or at least one cardiolipin.

4. The stimuli-sensitive nanocarrier system of claim 1, wherein R.sup.1 and R.sup.2 independently of one another are a linear saturated C12- to C24-alkyl functional group and m is 0 or 1.

5. The stimuli-sensitive nanocarrier system of claim 1, the system further comprising at least one phosphatidyldiglycerol and/or at least one phosphatidyltriglycerol.

6. The stimuli-sensitive nanocarrier system of claim 1 further comprising a phosphatidylcholine of formula (I) ##STR00034## wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms.

7. The stimuli-sensitive nanocarrier system of claim 1 comprising at least one phosphatidylcholine selected from the group consisting of 1-palmitoyl-2-oleoylglycero-3-phosphocholine, 1-stearoyl-2-oleoyl-3-phosphocholine, 1-palmitoyl-2-lauroylglycero-3-phosphocholine, 1-behenoyl-2-oleoylglycero-3-phosphocholine, 1-stearoyl-2-lauroylglycero-3-phosphocholine, 1,3-dimyristoylglycero-2-phosphocholine, 1,2-dimyristoylglycero-3-phosphocholine, 1-palmitoyl-2-myristoylglycero-3-phosphocholine, 1-stearoyl-2-myristoyl-glycero-3-phosphocholine, 1-myristoyl-2-palmitoylglycero-3-phosphocholine, 1,3-palmitoylglycero-2-phosphocholine, 1,2-dipalmitoylglycero-3-phosphocholine, 1-myristoyl-2 stearoylglycero-3-phosphocholine, 1-stearoyl-3 myristoylglycero-2-phosphocholine, 1-stearoyl-2-palmitoylglycero-3-phosphocholine, 1 palmitoyl-2-stearoylglycero-3-phosphocholine, 1,3-di stearoylglycero-2-phosphocholine, 1,2-distearoylglycero-3-phosphocholine, 1,2-diarachinoylglycero-3-phosphocholine, 1,2-dibehenoylglycero-3-phosphocholine and 1,2-dilignoceroylglycero-3-phosphocholine.

8. The stimuli-sensitive nanocarrier system of claim 1 comprising at least one phosphatidylcholine having a main transition temperature in the range of from 35 C. to 42 C. or in the range of from 40 C. to 43 C.

9. The stimuli-sensitive nanocarrier system of claim 8, wherein the at least one phosphatidylcholine is 1,3-dipalmitoyl-phosphatidylcholine or 1,2-dipalmitoylphosphatidylcholine.

10. The stimuli-sensitive nanocarrier system of claim 1, wherein the system does not comprise cholesterol.

11. A method for treating a tumor in an individual having the tumor comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

12. The method of claim 11 wherein the tumor is a soft-tissue sarcoma, osteosarcoma, bladder carcinoma, ovarian carcinoma, stomach carcinoma, breast carcinoma, hepatocellular carcinoma, uterine carcinoma, carcinoma of the thyroid gland, head-neck tumor, prostate carcinoma, chordoma, desmoid tumour, glioblastoma or other tumor disease.

13. The method of claim 11 wherein the tumor is a bladder tumor.

14. The method of claim 11 wherein the stimuli-sensitive nanocarrier system further comprises an active ingredient selected from the group consisting of anthracyclines, oxazaphosphorines, platinum analogues, gemcitabine, 5-fluorouracil, paclitaxel, docetaxel, etoposide, topotecan, vincristine, irinotecan, methotrexate, bleomycin, tyrosine kinase inhibitors, small molecules, DNA therapeutics and radiosensitisers.

15. The method of claim 11 wherein the stimuli-sensitive nanocarrier system further comprises a cytostatic selected from the group consisting of mitomycin C, doxorubicin, epirubicin, gemcitabine, trabectedin, cisplatin, carboplatin and oxaliplatin.

16. A method for treating infectious disease in an individual having the infectious disease comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

17. The method of claim 16 wherein the infectious disease is caused by bacteria, viruses, fungi and/or parasites.

18. The method of claim 16 wherein the infectious disease is an infection of a medical implant, a localized infection and/or an infection caused by multiresistant pathogens.

19. The method of claim 16, the stimuli-sensitive nanocarrier system further comprising an active ingredient selected from the group consisting of antibiotics, virostatics, fungicides and medicinal drugs having an anti-parasitic effect.

20. The method of claim 16, the stimuli-sensitive nanocarrier system further comprising an active ingredient selected from the group consisting of antibiotics, virostatics and fungicides.

21. A method for treating an individual having an eye disease comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

22. The method of claim 21, wherein the eye disease is an inflammatory disease, a degenerative disease, an infectious disease and/or a neoplastic disease, a wound healing disorder and/or glaucoma.

23. A method for treating an individual having an autoimmune disease comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

24. The method of claim 23, wherein the autoimmune disease is rheumatoid arthritis and/or a chronic inflammatory intestinal disease.

25. The method of claim 23, wherein the stimuli-sensitive nanocarrier system further comprises an active ingredient.

26. The method of claim 25, wherein the active ingredient is a steroid, TNF- and/or an immunosuppressant.

27. A method for performing a diagnosis comprising using the stimuli-sensitive nanocarrier system of claim 1.

28. The method of claim 27, further comprising using an MR contrast agent and MR imaging.

29. The method of claim 27, further comprising using an active ingredient.

30. The method of claim 29, wherein the active ingredient is a CT or MRT contrast agent.

31. A method for treating an individual having a degenerative disease comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

32. The method of claim 31, wherein the stimuli-sensitive nanocarrier system further comprises an active ingredient selected from the group consisting of anti-inflammatory agents, analgesics and/or chondroprotectants.

33. A method for treating an individual having dementia, Alzheimer's disease and/or a focal neurological psychiatric disease comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

34. A method for treating an individual having atherosclerosis comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

35. A method for treating an individual having a thrombosis comprising administration of the stimuli-sensitive nanocarrier system of claim 1 to the individual.

36. The method of claim 35, wherein the stimuli-sensitive nanocarrier system further comprises an active ingredient comprising a fibrinolytic.

37. The stimuli-sensitive nanocarrier system of claim 1, wherein the stimuli is hyperthermia and/or ultrasound.

38. A stereospecific phosphatidyloligoglycerol of formula (IIa) ##STR00035## wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms and m is an integer from 0 to 50, wherein the linkage from the glyceride to the phosphate group is stereospecific and is in the form of an sn-3 linkage and the linkage from the phosphate group to the oligoglycerol is stereospecific and is in the form of an sn-1 linkage.

Description

Example 1

(1) The Synthesis of Phosphatidyl-Oligoglycerols

(2) Important Structural Units

(3) 1) 3-Allyl-2-benzyl-sn-G (endogenous synthesis)

(4) 2) 1-Allyl-2-benzyl-sn-G (endogenous synthesis)

(5) 3) 1,2-Isopropylidene-sn-G (commercial product)

(6) 4) 2,3-Isopropylidene-sn-G (commercial product)

(7) By means of these structural units, the desired phosphatidyloligoglycerols can be developed, having a natural or also a non-natural configuration. Hitherto, only racemic phosphatidyloligoglycerols have been prepared and tested in long-term circulating liposomes in this field.

(8) Hitherto:

(9) ##STR00009##

(10) Configuration: racemic (non-natural)

(11) Name: 1,2-diacyl-sn-3-glycero-phospho-rac-diglycerol (correspondingly tri-OF tetra-glycerols)

(12) Explanations: R.sub.1, R.sub.2saturated alkyl functional groups; sn stereospecific numbering; racracemic linkage; correspondingly also tri- or tetra-glycerols

(13) According to the Invention:

(14) ##STR00010##

(15) Configuration: natural

(16) Name: 1,2-diacyl-sn-3-glycero-phospho-sn-1-diglycerol (correspondingly also tri- or tetra-glycerols)

(17) Explanations: R1, R2saturated alkyl functional groups; snstereospecific numbering; diglycerolscorrespondingly also tri- or tetra-glycerols

(18) The substances of natural configuration to the phosphate esters, that is to say (sn-3) linkage of diacylglycerol but (sn-1) linkage of the di-, tri- or tetra-glycerols, are described for the first time herein. They cannot be obtained from lecithins by transesterification with phospholipase D in the presence of glycerol, but only by targeted synthesis. The substances are in the natural configuration, that is to say (sn-3) and (sn-1) linkage.

(19) In the preparation of the further substances too, such as glycero-glycols glycero-glycerols (branched) glycero-glycero-glycols,

(20) synthesis routes have been developed which ensure that the substances contain a glycerol functional group which has an (sn-1) linkage of the phosphate ester to the phosphatidyloligoglycerol. These substances accordingly contain in the structure at least one glycerol molecule having a free OH group, which permits an (sn-1) linkage. The substances are novel, because an (sn-3) linkage of the diacyl-glycerolphosphoric acid ester and an (sn-1) linkage to the glycerol-glycol is possible here too. In addition, the structure here is also novel, because the terminal group is not glycerol but glycol or the like.

Example 2

(21) Examples of the Synthesis of Phosphatidyl-Oligoglycerols Having Uniform and Natural Configuration

(22) Abbreviations used; P, palmitic acid; O, oleic acid; S, stearic acid; L, lauric acid; B, behenic acid; M, myristic acid; A, arachidic acid; Li, linoceric acid,

(23) PC, phosphocholine; PG.sub.2, phosphodiglycerol; PG.sub.3, phosphotriglycerol; PG.sub.4, phosphotetraglycerol,

(24) ##STR00011##

Example 3

(25) Examples of the Synthesis of Phosphatidyl-Sn-1-Glycero Compounds of Natural Configuration (Novelty Through Structural Variation as Well as Through Uniform and Natural Configuration)

(26) Glycero-glycols (G-Gly)

(27) Structure:

(28) ##STR00012##

(29) Glycero-glycerols (branched)

(30) Structure:

(31) ##STR00013##

(32) Diglycero-glycos (G.sub.2-Gly)

(33) Structure:

(34) ##STR00014##

Example 4

(35) Use of Thermosensitive Liposomes in the Treatment of Bladder Carcinoma

(36) Thermosensitive liposomes according to the invention comprising 30 mol. % dipalmitoylphosphatidyldiglycerol or dipalmitoylphosphatidyltriglycerol were used.

(37) FIG. 1 shows the principle of the intravascular active-ingredient release from thermosensitive liposomes.

(38) Active-ingredient release generally occurs by passive transfer across the lipid membrane owing to a concentration gradient. At their phase transition temperature (T.sub.m), phospholipids transition from a solid gel phase (L.sub.) to a liquid disordered phase (L.sub.). The L.sub. phase is characterised by greater permeability compared with the L.sub. phase. The permeability is greatest at temperatures close to the phase transition temperature T.sub.m owing to the coexistence of membrane regions in which both phases occur, whereby boundary regions having packing defects occur. The thermosensitive liposomes according to the invention release their contents into the bloodstream when they pass through heated tissue. A high rate of active-ingredient release is thereby provided.

(39) In vivo experiments were carried out on female F344 rats weighing from 170 to 200 g which had developed an orthotopic bladder cancer through inoculation with AY27 cells. Pharmacokinetics and accumulation of doxorubicin (Dox) were evaluated by HPLC measurements. Focal tumour growth was initiated by chemical preconditioning of the bladder wall followed by AY27 cell instillation. Tumour growth was inspected by cystoscopy. TSL(Dox) (thermosensitive liposomes comprising doxorubicin) or free Dox were injected intravenously (i.v.) or introduced intravesically. Heating of the bladder was achieved by means of warm water.

(40) FIG. 2 shows the pharmacokinetic profile of TSL(Dox).

(41) A TSL(Dox) administration of 2 mg/kg was carried out on female F344 rats on day 0 with a repeat administration after 7 or 14 days. TSL(Dox) exhibit high stability over 120 minutes.

(42) FIG. 3 shows the experimental design of the rat bladder carcinoma model (modified by Postius, Szelinyi; J. Pharmacol. Methods. 1983, 9: 53-61).

(43) The rat is provided with two bladder catheters (1). A precision pump (2) transports heated water from the water bath (3) through one of the catheters into the bladder. The other catheter serves to discharge the warm water. When the temperature of the outgoing water reaches 41 C., thermosensitive liposomes comprising doxorubicin are injected into the tail vein and a one-hour hyperthermia treatment is started.

(44) FIG. 4 shows a tumour macroscopy seven days after tumour cell inoculation.

(45) The experiments were carried out on female F344 rats weighing from 170 to 200 g which had developed an orthoptic bladder cancer through inoculation with AY cells. Pharmacokinetics and accumulation of doxorubicin (Dox) was evaluated by HPLC measurements. Focal tumour growth was initiated by chemical preconditioning of the bladder wall followed by AY27 cell instillation. Multiple tumour plaque formations on the bladder wall were visualised by cytoscopy (A) and on a removed bladder (B and C).

(46) FIG. 5 shows the histology of a bladder.

(47) FIG. (A) shows a bladder of normal appearance of an F344 rat at 10 magnification. FIG. (B) shows a pT1G3 tumour seven days after tumour cell inoculation, likewise at 10 magnification.

(48) FIG. 6 shows tissue concentrations of Dox in the bladder wall of female F344 rats having a bladder tumour.

(49) The figure shows the mean Dox concentration in the urothelium and in the muscle layer after treatment for one hour with intravenously administered TSL(Dox) and hyperthermia, intravesically administered free Dox (fDOX) and intravenously administered free Dox. The Dox dose for the i.v. treatment was 5 mg/kg body weight, and for the intravesical treatment it was 0.5 mg/rat. The Dox concentration in the urothelium was higher in all cases, regardless of the treatment procedure. The concentration in the urothelium/in the muscle layer of a tumour-bearing rat which had been treated with TSL(Dox)+hyperthermia was up to 3- to 5-times higher in comparison with intravesically administered free Dox.

Example 5

(50) Important Structural Units for the Synthesis of Phosphatidyl-Diglycerols and Analogues

A) (sn)-1,2-Isopropylideneglycerol

(51) is a commercial product having a free (sn)-3-hydroxyl group. It can be used directly

(52) ##STR00015## for phosphorylation. Phosphoric acid esters having (sn)-3 linkage are formed:

B) (sn)-3-Allylglycerol

(53) can be obtained from A by reaction with alkyl chloride and then by acidic cleavage of the isopropylidene protecting group. It serves to synthesise optically pure glycerol derivatives which have a free hydroxyl group in the (sn)-1 position.

C) (sn)-1-Trityl-2-benzyl-3-allyl-glycerol

(54) can be obtained from B by tritylation in (sn)-1 and subsequent benzylation in (sn-2).

D) (sn)-1-Trityl-2-benzyl-3-glycero-glycerol

(55) can be developed from C by epoxidation and ring opening.

E) (sn)-2-Benzyl-3-glycero-dibenzyl-glycerol

(56) can be prepared from D by dibenzylation and subsequent detritylation. There is formed an important structural unit which permits (sn)-1 linkage and thus the preparation of (sn)-1-phosphoric acid esters.

(57) ##STR00016##

F) (sn)-1-OH-2-Benzyl-3-glycero-benzyl-glycol

(58) can be developed from D: vicinal diol cleavage using periodate and reduction of the aldehyde to the alcohol using sodium borohydride yields 1-trityl-2-benzyl-3-glycero-glycol. Benzylation of the free hydroxyl group and detritylation lead to the freeing of the (sn)-1 position, which can again be used for the preparation of (sn)-1-phosphoric acid esters.

(59) ##STR00017## In a corresponding manner, (sn)-1-OH-2-benzyl-3-glycero-2-benzyl-glycero-benzyl-glycol can be developed from triglycerols using the same protecting group strategy:

(60) ##STR00018##

G) (sn)-1-Acetyl-2-benzyl-glycerol

(61) this protecting group is only used to prepare complex phospholipids such as cardiolipin, which contains two phosphoric acid esters in one molecule. The phosphatidyl functional groups present in the molecule are bonded together via a glycerol bridge. The above structural unit allows the production of this bridge as well as the preparation of the two phosphoric acid esters in the (sn)-3 position, that is to say likewise in the naturally occurring configuration. G can be developed from structural unit C: detritylation in the (sn)-1 position, rearrangement of allyl to propenyl, acetylation of the (sn)-1 position and acidic cleavage of the propenyl protecting group yields structural unit G having a free (sn)-3 position:

(62) ##STR00019## For the preparation of cardiolipins and analogues, particular measures must be taken because two negative charge carriers, that is to say two phosphate functional groups per molecule, are present in these molecules. To that end, the already phosphorylated functional group R.sub.5 is used, after cleavage of the acetyl group, as the alcohol R.sub.6 OH (see in this connection preparation of the phosphoric acid triesters: preparation of the phosphoric acid triester from R.sub.5OH and preparation of the phosphoric acid triester from R.sub.6OH)

(63) ##STR00020## If R.sub.6OH is again reacted with phosphorus oxychloride, the diphosphate is obtained. The two phosphoric acid functional groups are then bonded together via a glycerol bridge.

(64) ##STR00021## At the end of the synthesis, the four hydroxyl groups protected by isopropylidene are then freed. Acylation is then carried out, as is conventional, by known methods fatty acid chloride, for example palmitic acid chloride or palmitic acid. After cleavage of the benzyl ether protecting groups by catalytic hydrogenic hydrogenolysis with PD-C, the target product is obtained by cleavage of the methyl group using lithium bromide.

1-Acetylbropanediol-(1,3)

(65) can likewise be used as the bridging molecule and then yields the deoxy-cardiolipinslikewise with other terminal diols such as glycol, (1,4)-butanediol, etc. These compounds are of lesser interest in our concept, which intends to use preferably naturally occurring phosphatidyl derivatives.

Example 6

(66) Preparation of Strategically Important Phosphoric Acid Triesters by Stepwise Reaction of POCl.sub.3 with Three Different, Primary Alcohols

(67) The preparation of defined, position-specific phosphoric acid triesters which are clear in terms of configuration has been researched intensively (see in this connection publications 1-10). 1) Eibl H. Synthesis of glycerophospholipids. Chem Phys Lipids. 1980 June; 26(4): 405-29 2) Eibl H. Phospholipid synthesis. In: Liposomes: From physical structure to therapeutic applications. Ed Knight C G, Elsevier, Amsterdam 1981; 19-50 3) Eibl H, Kovatchev S. Preparation of phospholipids and their analogs by phospholipase D. Methods Enzymol. Ed Lwenstein J M. 1981; 72: 632-9. Academic Press, New York 4) Eibl H. Phospholipide als funktionelle Bausteine biologischer Membranen. Angew Chem. 1984 (259): 9188-9198 5) Eibl H. Phospholipids as functional constituents of biomembranes. Angew Chem Int. Ed Engl 1984 (23) 257-271 6) Eibl H. Phospholipid synthesis: Oxazaphospholanes and dioxaphospholanes as intermediates. Proc Natl Acad Sci USA. 1978; 75: 4074-77 7) Eibl H, Woolley P. Synthesis of enantiomerically pure glyceryl esters and ethers. I. Methods employing the precursor 1,2-isopylidene-sn-glycerol. Chem Phys Lipids 1986 (41): 53-63 8) Eibl H, Woolley P. Synthesis of enantiomerically pure glyceryl esters and ethers. II. Methods employing the precursor 3,4-isopropylidene-D-mannitol. Chem Phys Lipids. 1988 (47): 47-53 9) Eibl H, Woolley P. A general synthetic method for enantiomerically pure ester and ether lysophospholipids. Chem Phys Lipid 1988 (47): 63-68 10) Woolley P. Eibl H. Synthesis of enantiomerically pure phospholipids including phosphatidylserine and phosphatidylglycerol. Chem Phys Lipids 1988 (47): 55-62

(68) From the structural units described here, it is possible to prepare different phosphatidyl-oligo-glycerols and, correspondingly, phosphatidyl-glycero-glycols. It is particularly important that the fatty acid functional groups are prepared only at the end of the synthesis from a central intermediate, that is to say, unlike previous syntheses, the fatty acid functional groups are not introduced until the end. This is possible by preparing a central intermediate having two vicinal hydroxyl groups, for example

(69) ##STR00022##

(70) In German patent application 196 05 833.3 of 16 Feb. 1996 (inventor: H. Eibl; patentee: Max-Planck-Gesellschaft), the conditions for the stepwise esterification of phosphorus oxychloride using primary alcohols are discussed and described precisely. In contrast to the previous processes, in which 1,2-diacylglycerol, for example 1,2-dipalmitoylglycerol, was used in the first step (problem: fatty acid migration during the reaction or during storage of 1,2-dipalmitoylglycerol, which had to be prepared separately beforehand), the synthesis is initiated with (sn)-1,2-isopropylideneglycerol having a free sn-3 position, which can be obtained commercially. This saves the complex, separate synthesis of 1,2-diacylglycerol, for example 1,2-dipalmitoylglycerol, and the above-mentioned problems associated therewith.

(71) Following these poor experiences with 1,2-dipalmitoyl-glycerol in the first phosphorylation step, the synthesis was carried out differently. The aim was to introduce the fatty acid esters only at the end of the synthesis via a freed vicinal diol. The following sequence of the synthesis steps is preferable:

(72) P.sub.1first phosphorylation step with phosphorus oxychloride R.sub.1 OH: (sn)-1,2-isopropylidene-glycerol (free sn-3 position)
POCl.sub.3+R.sub.1 OH.fwdarw.R.sub.1 OPO Cl.sub.2

(73) P.sub.2second phosphorylation step with R.sub.1 OPO Cl.sub.2 R.sub.2 OH or corresponding having free sn-1 position R.sub.3 OH; R.sub.4 OH; R.sub.5 OH; R.sub.6 OH

(74) ##STR00023##

(75) P.sub.3third phosphorylation step with

(76) ##STR00024## Methanolysis with CH.sub.3 OH

(77) ##STR00025##

(78) Furthermore, after phosphorylation using the benzyl-protected glycerols, the phosphoric acid diesters are not hydrolysed directly but are converted into phosphoric acid triesters by methanolysis. Freeing of the vicinal diol can then take place by acidic cleavage of the isopropylidene protecting group. The acylation of the hydroxyl groups is then carried out according to known methods (fatty acid chloride, for example palmitic acid chloride, or also free fatty acid).

(79) The target products are then obtained by PD/C-catalysed hydrogenolysis followed by methyl cleavage using lithium bromide.

(80) By way of example, some target products will be shown in graphic formulae, for example as dipalmitoyl esters:

(81) Phosphatidyldiglycerols and Analogues

(82) Phosphatidyl-diglycerols and phosphatidyl-triglycerols have been developed from phosphatidylglycerol, a naturally occurring membrane phospholipid. Surprisingly, the introduction of a further glycerol functional group which is linked via an ether bridge to phosphatidylglycerol leads to particular properties: The circulation times in the blood of liposomes comprising this phospholipid are changed and lengthened considerably.

(83) ##STR00026##

(sn)-1,2-Dipalmitoyl-glycero-3-phospho-sn-1-glycero-glycerol, sodium salt

(84) ##STR00027##

(sn)-1,2-Dipalmitoyl-sn-glycero-3-phospho-glyceroglycol, sodium salt

(85) Cardiolipins and Structural Analogues

(86) Cardiolipins and analogues, which likewise occur in natural membranes, also have properties that change the circulation time, and increase the circulation time of liposomes in the blood.

(87) Using the structural units developed by us, it is also possible to prepare mixed-chain cardiolipins, that is to say, for example, also structures having only three fatty acid functional groups.

(88) ##STR00028##

(sn)-1,2-Dipalmitoyl-3-glycero-phospho-glycero-(sn)-3-phosphoglycerol, disodium salt

7. Examples of (sn)-1,2-diacyl-3-glycero-phospho-(sn)-1-diglycerol

(89) 1) Dipalmitoyl compound sodium salt; C.sub.41 H.sub.80 Na O.sub.12 P (MW 819.04) 2) Dimyristoyl compound sodium salt; C.sub.37 H.sub.72 Na O.sub.12 P (MW 762.93) 3) Distearoyl compound sodium salt; C.sub.45 H.sub.88 Na O.sub.12 P (MW 875.14) 4) 1-Palmitoyl-2-lauroyl compound sodium salt; C.sub.37 H.sub.72 Na O.sub.12 P (MW 762.93) 5) 1-Stearoyl-2-lauroyl compound sodium salt; C.sub.39 H.sub.76 Na O.sub.12 P (MW 790.98) 6) 1-Stearoyl-2-myristoyl compound sodium salt; C.sub.41 H.sub.80 Na O.sub.12 P (MW 819.04) 7) 1-Stearoyl-2-palmitoyl compound sodium salt; C.sub.43 H.sub.84 Na O.sub.12 P (MW 847.09)

8. Examples of (sn)-1,2-diacyl-3-glycero-phospho-(sn)-1-glycero-glycol

(90) 1) Dipalmitoyl compound sodium salt; C40 H80 Na O12 P (MW 789.04) 2) Distearoyl compound sodium salt; C44 H86 Na O12 P (MW 845.14) 3) 1-Stearoyl-2-myristoyl compound sodium salt; C40 H78 Na O12 P (MW 789.04) 4) 1-Stearoyl-2-palmitoyl compound sodium salt; C42 H82 Na O12 P (MW 817.09)

9. Examples of (sn)-1,2-diacyl-3-glycero-phospho-(sn)-1-diglycero-glycol

(91) 1) Dipalmitoyl compound sodium salt; C43 H84 Na O12 P (MW 863.12) 2) Distearoyl compound sodium salt; C.sub.47 H.sub.90 Na O.sub.12 P (MW 919.22)

10. Examples of (sn)-1,2-diacyl-3-glycero-phospho-(sn)-1-triglycerol

(92) 1) Dipalmitoyl compound sodium salt; C44 H86 Na O12 P (MW 893.12) 2) Distearoyl compound sodium salt; C48 H92 Na O12 P (MW 949.22) Item 1 Stimuli-sensitive nanocarrier system. Item 2 Stimuli-sensitive nanocarrier system for use in locoregional therapy. Item 3 Stimuli-sensitive nanocarrier system according to either item 1 or item 2, characterised in that said system is a thermosensitive liposome. Item 4 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 3, characterised in that said system comprises (i) at least one phosphatidylcholine having a main transi ion temperature of from 0 C. to 80 C. and (ii) at least one phosphatidyloligoglycerol and/or at least one phosphatidylglyceroglycol and/or at least one cardiolipin. Item 5 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 4, characterised in that said system comprises at least one phosphatidyloligoglycerol of formula (II)

(93) ##STR00029## wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms and n is an integer from 2 to 50. Item 6 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 4, characterised in that said system comprises at least one phosphatidyloligoglycerol of formula (IIa)

(94) ##STR00030## wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms and m is an integer from 0 to 50, wherein the linkage from the glyceride to the phosphate group is stereospecific and is in the form of an sn-3 linkage and the linkage from the phosphate group to the oligoglycerol is stereospecific and is in the form of an sn-1 linkage. Item 7 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to either item 5 or item 6, characterised in that R.sup.1 and R.sup.2 independently of one another are a linear saturated C12- to C24-alkyl functional group and n is 2 or 3 or m is 0 or 1. Item 8 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 4 to 7, characterised in that said system comprises at least one phosphatidyldiglycerol and/or at least one phosphatidyltriglycerol. Item 9 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 8, comprising a phosphatidylcholine of formula (I)

(95) ##STR00031## wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms. Item 10 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of the preceding items, characterised in that said system comprises at least one phosphatidylcholine selected from the group consisting of 1-palmitoyl-2-oleoylglycero-3-phosphocholine, 1-stearoyl-2-oleoyl-3-phosphocholine, 1-palmitoyl-2-lauroylglycero-3-phosphocholine, 1-behenoyl-2-oleoylglycero-3-phosphocholine, 1-stearoyl-2-lauroylglycero-3-phosphocholine, 1,3-dimyristoylglycero-2-phosphocholine, 1,2-dimyristoylglycero-3-phosphocholine, 1-palmitoyl-2-myristoylglycero-3-phosphocholine, 1-stearoyl-2-myristoylglycero-3-phosphocholine, 1-myristoyl-2-palmitoylglycero-3-phosphocholine, 1,3-palmitoylglycero-2-phosphocholine, 1,2-dipalmitoylglycero-3-phosphocholine, 1-myristoyl-2-stearoylglycero-3-phosphocholine, 1-stearoyl-3-myristoylglycero-2-phosphocholine, 1-stearoyl-2-palmitoylglycero-3-phosphocholine, 1-palmitoyl-2-stearoylglycero-3-phosphocholine, 1,3-distearoylglycero-2-phosphocholine, 1,2-distearoylglycero-3-phosphocholine, 1,2-diarachinoylglycero-3-phosphocholine, 1,2-dibehenoylglycero-3-phosphocholine and 1,2-dilignoceroylglycero-3-phosphocholine. Item 11 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of the preceding items, characterised in that said system comprises at least one phosphatidylcholine having a main transition temperature in the range of from 35 C. to 42 C. or in the range of from 40 C. to 43 C. Item 12 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to either item 10 or item 11, characterised in that said system comprises at least one phosphatidylcholine selected from 1,3-dipalmitoylphosphatidylcholine and 1,2-dipalmitoylphosphatidylcholine. Item 13 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of the preceding items, characterised in that it does not comprise cholesterol. Item 14 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of the preceding items for use in the treatment of tumours. Item 15 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 14 for use in the treatment of soft-tissue sarcoma, osteosarcoma, bladder carcinoma (muscle invasive bladder cancer [MIBC] and non-muscle invasive bladder cancer [NMIBC]), ovarian carcinoma, stomach carcinoma, breast carcinoma (especially triple negative breast cancer [TNBC]), hepatocellular carcinoma, uterine carcinoma, carcinoma of the thyroid gland, head-neck tumours, prostate carcinoma, chordoma, desmoid tumour, glioblastoma and other tumour diseases having preferably locoregional spread. Item 16 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to either item 14 or item 15 for use in the treatment of bladder tumours. Item 17 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 14 to 16, further comprising an active ingredient, in particular selected from anthracyclines (for example doxorubicin, epirubicin), oxazaphosphorines (for example hydroxyifosfamide), platinum analogues (cisplatin, oxaliplatin, carboplatin), gemcitabine, 5-fluorouracil, paclitaxel, docetaxel, etoposide, topotecan, vincristine, irinotecan, methotrexate, bleomycin, tyrosine kinase inhibitors, small molecules, DNA therapeutics, radiosensitisers (in conjunction with radiotherapy). Item 18 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of infectious diseases. Item 19 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 18, characterised in that the infectious disease is caused by bacteria, viruses, fungi and/or parasites. Item 20 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to either item 18 or item 19 for the treatment of infections of medical implants, in particular orthopaedic prostheses, for the treatment of localised infections, in particular infections of the deep soft tissue and/or of the bone, and/or for the therapy of multiresistant pathogens. Item 21 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 18 to 20, further comprising an active ingredient, in particular selected from antibiotics, virostatics, fungicides, and medicinal drugs having an anti-parasitic effect. Item 22 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 18 to 21, characterised in that the active ingredient is selected from antibiotics, in particular -lactams, glycopeptides, polyketides, aminoglycoside antibiotics, polypeptide antibiotics, quinolones, sulfonamides (for example linezolid, flucloxacillin, cefazolin, clindamycin, vancomycin, teicoplanin, rifampicin, ampicillin, ceftazidime, ceftriaxone, cefepime, piperacillin, fluoroquinolones, metronidazole, amikacin, etc.) and/or virostatics, in particular entry inhibitors, penetration inhibitors, DNA polymerase inhibitors, DNA/RNA polymerase inhibitors, reverse transcriptase inhibitors, inosine monophosphate dehydrogenase inhibitors, protease inhibitors, integrase inhibitors, helicase-primase inhibitors, cyclophilin inhibitors, maturation inhibitors, terminase inhibitors, neuraminidase inhibitors, etc., and/or fungicides, in particular azoles (benzimidazoles (MBC), triazoles, imidazoles), morpholines, strobilurins, quinolines, anilino-pyrimidines, oxazolidine-diones, carboxylic acid amides, etc. Item 23 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of diseases of the eye. Item 24 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 23 for the treatment of inflammatory, degenerative, infectious and/or neoplastic diseases of the eye, wound healing disorders and/or glaucoma. Item 25 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of autoimmune diseases. Item 26 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 25 for use in the treatment of rheumatoid arthritis and/or chronic inflammatory intestinal diseases. Item 27 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to either item 25 or item 26, further comprising an active ingredient, in particular a steroid, TNF- and/or immunosuppressants. Item 28 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in diagnosis. Item 29 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 28 for non-invasive temperature measurement using MR contrast agents and MR imaging. Item 30 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to either item 28 or item 29, further comprising an active ingredient, in particular a CT or MRT contrast agent, preferably selected from iodine-containing contrast agents or gadolinium chelates. Item 31 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of degenerative diseases. Item 32 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 31 for the localised release of anti-inflammatory agents, analgesics and/or chondroprotectants. Item 33 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of dementia syndromes, Alzheimer's disease and/or focal neurological psychiatric diseases, in particular epilepsy. Item 34 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of atherosclerosis. Item 35 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of items 1 to 13 for use in the treatment of thromboses. Item 36 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 35, further comprising at least one active ingredient, in particular selected from fibrinolytics, preferably streptokinase, urokinase and/or alteplase. Item 37 Stimuli-sensitive nanocarrier system according to any of the preceding items, wherein the stimuli-sensitive nanocarrier system is changed and in particular opened by the exertion of a stimulus in order to release from the nanocarrier system an active ingredient which may be contained therein. Item 38 Stimuli-sensitive nanocarrier system according to item 37, wherein the stimulus is selected from radio frequency (for example radiative superficial and deep hyperthermia systems, bladder hyperthermia systems), ultrasound (for example highly focused ultrasound [high intensity focused ultrasound, HIFU], low intensity ultrasound [low intensity focused ultrasound, LIFU]), light, laser, conduction through heated liquid, other physical principles which either lead to locoregional heating and/or can destabilise membranes consisting of phospholipids. Item 39 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of the preceding items 14 to 16, characterised in that said system further comprises a cytostatic. Item 40 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to item 39, characterised in that the cytostatic is selected from the group consisting of mitomycin C, doxorubicin, epirubicin, gemcitabine, trabectedin, cisplatin, carboplatin and oxaliplatin. Item 41 Stimuli-sensitive nanocarrier system, in particular thermosensitive liposome, according to any of the preceding items in combination with hyperthermia and/or ultrasound. Item 42 Stereospecific phosphatidyloligoglycerol of formula (IIa)

(96) ##STR00032## Configuration: natural Name: 1,2-diacyl-sn-3-glycero-phospho-sn-1-oligoglycerol; sn, stereospecific numbering; wherein R.sup.1 and R.sup.2 each independently represent a hydrocarbon functional group having from 12 to 24 carbon atoms and m is an integer from 0 to 50, wherein the linkage from the glyceride to the phosphate group is stereospecific and is in the form of an sn-3 linkage and the linkage from the phosphate group to the oligoglycerol is stereospecific and is in the form of an sn-1 linkage. Item 43 Stimuli-sensitive liposome comprising a stereospecific phosphatidyloligoglycerol of formula (IIa) according to item 42 for use in the treatment of bladder tumours. Item 44 Stimuli-sensitive liposome according to item 43, further comprising a phosphatidylcholine of formula (I) having a main transition temperature of from 35 to 40 C., in particular from 40 to 43 C.