AU2013203682B2 - Liposome comprising at least one cholesterol derivative - Google Patents

Abstract

Abstract Liposome comprising at least one cholesterol derivative. 5 The present invention relates to a novel galenic form of cholesterol derivative. More particularly the invention relates to liposomes comprising at least one cholesterol derivative and compositions comprising said liposomes.

Description

1 Liposome comprising at least one cholesterol derivative. The present invention relates to a novel galenic form of cholesterol derivatives. More particularly, the invention relates to liposomes comprising at least one cholesterol derivative corresponding to formula (I) described hereafter and the 5 compositions comprising said liposomes. By way of simplification, the expression "cholesterol derivative" in the present text can be used to denote the compounds of formula (I), or even their homologues which are not specifically described in the present Application, as well as their analogues. In fact, all these compounds of formulae I have the cholesterol skeleton 10 in common. The invention relates to, but is not constituted only by, all these compounds and their homologues not mentioned here, which have structural similarities. According to a first aspect of the present invention there is provided a liposome comprising at least one compound corresponding to the following formula (I) R6 R7 R8 R1 H H H X R5 R4 15 z R2 R3 in which,

R

1 represents a hydrogen atom or a -CH 3 , -CH 2 -CN, -CH 2 -ORa, -CH 2 -SRa, -CH 2 SeRa, -C(O)-Ra, -C(O)ORa, -O-C(O)NRaR', -C(O)NRaR6 group, in which (i) Ra and Rb, simultaneously or independently of one another, are chosen 20 from a hydrogen atom or a C 1

-C

6 alkyl group, a C 2

-C

6 alkenyl group, a C 3 C 6 cycloalkyl group, an aryl group, a heterocyclic group or (ii) Ra and R6 together form a linear or branched hydrocarbon chain, having 2 to 6 carbon atoms, optionally comprising one or more double bonds and/or optionally interrupted by one or more oxygen, sulphur or nitrogen 25 atom(s), or la (iii) Ra and Rb together with the nitrogen to which they are attached form a C 3 C 6 heterocycle, said heterocycle can comprise one or more double bonds and/or one or more oxygen, sulphur or nitrogen atom(s); * R 2 represents a hydrogen atom or a C 1

-C

6 alkyl, C 3

-C

6 cycloalkyl, C 2

-C

6 alkenyl, 5 C 2

-C

6 alkynyl, aryl, a heterocycle group, or a halogen atom or a -CN, -CF 3 , -NO 2 , ORa, -SRa, -SO 2 Ra, -NRaR6, -C(O)-Ra, -OC(O)Ra, -OC(O)NRaR6, -C(O)ORa _ C(O)NRaRb group, Ra and Rb being as defined previously * R 3 represents a hydrogen atom or a C 1

-C

6 alkyl group, or 10 * R 2 and R 3 together with the carbon to which they are attached, represent an oxygen atom or a C 3

-C

6 cycloalkyl group or an =N-OH, =CH-(C 1

-C

6 )alkyl, =CH aryl, =CH-(C 3

-C

6 )cycloalkyl group; * R 4 represents a hydrogen atom or a C 1

-C

6 alkyl group, or a hydroxyamino (-NH 2 OH) group; or 15 * R 3 and R 4 together form an additional carbon-carbon bond between the carbon atoms to which they are attached, or a C 3

-C

6 cycloalkyl group; * R 5 represents a hydrogen atom or a C 1

-C

6 alkyl, C 3

-C

6 cycloalkyl, C 2

-C

6 alkenyl,

C

2

-C

6 alkynyl, aryl group, or a -CN, -ORa, -SRa, -SO 2 Ra, -NRaR6, -C(O)-Ra OC(O)Ra, -OC(O)NRaR6 group, Ra and Rb being as defined previously, or a 20 hydroxyamino (-NH 2 -OH) group; * R 8 represents a group chosen from (i) a C 4

-C

12 alkyl group or a C 4

-C

12 alkenyl group, in particular a group chosen from 25 (ii) a group corresponding to formula (II) as follows:

R

14

-Y-R

1 5 (II) in which: lb - R 14 represents a C 4

-C

12 alkyl group or a C 4

-C

12 alkenyl group, in particular a C 5 C 10 alkyl group, preferentially the following G 7 group 20 25 y and - Y represents an oxygen atom or an -NRa group with Ra being as described 5 previously, and - R 15 represents a C 1

-C

6 alkyl, C 3

-C

6 cycloalkyl, aryl, heterocycle, -C(O)-(C 1 C 6 )alkyl, -C(O)-(C 3

-C

6 )cycloalkyl, -C(O)-aryl, -C(O)-heterocycle group, in particular a group represented by one of formulae (Ill) or (IV) 0 HN NH H H 0 S

NO

2 (111) (IV) 10 " R 6 represents a hydrogen atom or a halogen atom or a hydroxyl group, preferentially a hydrogen atom; " R 7 represents a hydrogen atom, or an -ORa group, Ra being as defined previously, preferentially a hydrogen atom; 15 and in which, * X and Z together with the carbon to which they are attached are a ketone (=0) group, an (=N-OH) oxime group or an alkyl (=N-0-alkyl) oxime group the alkyl of which group can be C 1

-C

6 * A represents 20 - a hydrogen atom, or - a C 1

-C

8 alkyl group, or - a C 2

-C

8 alkenyl, or 1c - a C 2

-C

8 alkynyl, or - a C 3

-C

6 cycloalkyl, or - an aryl, or - a heterocycle, or 5 - a halogen atom or - a -(CH 2 )n-CN, -(CH 2 )n-CF 3 , -(CH 2 )n-NO 2 , -(CH 2 )n-ORa, -(CH 2 )n-C(Me) 2 ORa _

(CH

2 )n-CHMeORa,-(CH 2 )n-SRa, -(CH 2 )n-SO 2 Ra, -CH 2 -SeRa, -(CH 2 )n-NRaR6, C(O)Ra, -OC(O)NRaR6, -C(O)ORa, -C(O)NRaR6 group in which (i) Ra and Rb, are as defined previously and 10 (ii) n represents an integer which can have any one of the values from 0 to 4, or also - a group corresponding to formula (V): Rc-Q-(CH 2 )m (V) in which 15 (i) m represents an equal integer which can have any one of the values from 1 to 8; and (ii) Q represents an oxygen atom or an -NRa group in which Ra is as defined previously and (iii) Rc represents 20 a. a hydrogen atom or b. a C 1

-C

6 alkyl, c. an aryl, d. a heteroaryl, e. a heterocycle, 25 f. an alkyl-C(O)-, the alkyl of which can be C 1

-C

6 , g. an aryl-C(O)-, h. a heteroaryl-C(O)-, i. a heterocycle-C(O)-, j. a group represented by one of formulae (III) or (IV) 1d 0 HN NH H H 0 S NO 2 (111) (IV) or k. an -O-C(O)- group or I. an -NRa-C(O)- group in which Ra is as defined previously 5 as well as: - its SYN, ANT/geometrical isomers, when they exist, - its optical isomers (enantiomers, diastereoisomers), when they exist, - its addition salts with a pharmaceutically acceptable acid or base, - its hydrates and its solvates, 10 - its prodrugs, - or one of its esters. According to a second aspect of the present invention there is provided a composition comprising at least one liposome comprising at least one compound of formula (I) as described in the first aspect of the present invention. 15 According to a third aspect of the present invention there is provided use of at least one compound of formula (I) as described in the first aspect of the present invention for preparing liposomes. According to a fourth aspect of the present invention there is provided use of the liposomes according to the first aspect of the present invention, for preparing a 20 composition, advantageously a weakly reactogenic, cosmetic, pharmaceutical, or veterinary composition. According to a fifth aspect of the present invention there is provided use of the liposomes according to the first aspect of the present invention, for the transport of active ingredients other than the compounds of formula (I). 25 Thus the invention relates to liposomes comprising at least one compound corresponding to the following formula (I) le R6 R7 R8 R1 H H H X R5 R4 Z R2 R3 in which, * R 2 can represent a hydrogen atom or a C 1

-C

6 alkyl, C 3

-C

6 cycloalkyl, C 2

-C

6 alkenyl, C 2

-C

6 alkynyl, aryl group, a heterocycle, or a halogen atom or a -CN, 5 -CF 3 , -NO 2 , -ORa, -SRa, -SO 2 Ra, -NRaRb, -C(O)-Ra, -OC(O)Ra, -OC(O)NRaR, -C(O)ORa, -C(O)NRaRb group, in which (i) Ra and Rb, simultaneously or independently of one another, can be chosen from a hydrogen atom or a C 1

-C

6 alkyl group, a C 2

-C

6 alkenyl 2 group, a C3-C6 cycloalkyl group, an aryl group, a heterocyclic group or (ii) Ra and Rb together can form a linear or branched hydrocarbon chain, having 2 to 6 carbon atoms, optionally comprising one or more 5 double bonds and/or optionally interrupted by one or more oxygen, sulphur or nitrogen atom(s), or (iii) Ra and Rb together with the nitrogen to which they are attached can form a C3-C6 heterocycle, said heterocycle can comprise one or more double bonds and/or one or more oxygen, sulphur or nitrogen 10 atom(s); " R 3 can represent a hydrogen atom or a C 1

-C

6 alkyl group, or " R 2 and R 3 together with the carbon to which they are attached, can represent an oxygen atom or a C3-C6 cycloalkyl group or an =N-OH, =CH-(C 1

-C

6 )alkyl, =CH aryl, =CH-(C 3

-C

6 )cycloalkyl group; 15 e R 4 can represent a hydrogen atom or a C1-C6 alkyl group, or a hydroxyamino

(-NH

2 -OH) group; or " R 3 and R 4 together can form an additional carbon-carbon bond between the carbon atoms to which they are attached, or a C3-C6 cycloalkyl group; " R 5 can represent a hydrogen atom or a C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 20 alkenyl, C2-C6 alkynyl, aryl group, or a -CN, -ORa, -SRa, -S0 2 R', -NRaRb, -C(O) Ra, -OC(O)Ra, -OC(O)NRaRb group, Ra and Rb being as defined previously, or a hydroxyamino (-NH 2 -OH) group; " R 8 can represent a group chosen from (i) a C4-C12 alkyl group or a C4-C12 alkenyl group, in particular a group chosen 25 from / iG2G G4 G5

G

3 (ii) a group corresponding to formula (II) as follows:

R

14

-Y-R

15 (II) in which: - R 14 can represent a C4-C12 alkyl group or a C4-C12 alkenyl group, in 5 particular a C5-Cia alkyl, group preferentially the following G 7 group 20 Y 7 and - Y can represent an oxygen atom or an -NRa group with Ra being as described previously, and - R 1 5 can represent a CrC6 alkyl, C3-C6 cycloalkyl, aryl, heterocycle, 10 -C(O)-(Cr 1

C

6 )alkyl, -C(O)-(C 3

-C

6 )cycloalkyl, -C(O)-aryl, -C(O)-heterocycle group, in particular a group represented by one of formulae (111) or (IV) 0 HN NH r \H' ... .. H 0 0 S

NO

2 (Ill) (IV) * R 6 can represent a hydrogen atom or a halogen atom or a hydroxyl group, preferentially a hydrogen atom; 15 * R 7 can represent a hydrogen atom, or an -ORa group, R' being as defined previously, preferentially a hydrogen atom; it being understood that o If X and Z together with the carbon to which they are attached represent a ketone (=0) group, an (=N-OH) oxime group or an alkyl (=N-0-alkyl) oxime 20 group the alkyl of which group can be Cr1C6 then 4 0 R 1 can represent a hydrogen atom or a -CH 3 , -CH 2 -CN,

-CH

2 -ORa, -CH 2 -SRa, -CH 2 -SeRa, -C(O)-Ra, -C(O)ORa , -O-C(O)NRaR, -C(O)NRaRb group, R' and Rb being as defined previously, * A can represent 5 - a hydrogen atom, or - a C-C alkyl group, or - a C2-Ca alkenyl, or - a C2-Ca alkynyl, or - a C3-C6 cycloalkyl, or 10 - an aryl, or - a heterocycle, or - a halogen atom or - a-(CH 2 )n-CN, -(CH 2 )n-CF 3 , -(CH 2 )n-NO 2 , -(CH 2 )n-ORa

-(CH

2 )n-C(Me) 2 ORa, (CH 2 )n-CHMeORa,_(CH 2 )n-SRa _(CH 2 )n-SO 2 R a 15 -CH 2 -SeRa, -(CH 2 )n-NRaR, -C(O)Ra, -OC(O)NRaR', -C(O)ORa -C(O)NRaR group in which (i) Ra and Rb, are as defined previously and (ii) n can represent an equal integer which can have any one of the values from 0 to 4, or also 20 - a group corresponding to formula (V):

RC-Q-(CH

2 )m (V) in which (i) m can represent an integer which can have any one of the values from 1 to 8; and 25 (ii) Q can represent an oxygen atom or an -NRa group in which Ra is as defined previously and (iii) Rc can represent a. a hydrogen atom or b. a C-C 6 alkyl, 30 c. an aryl, d. a heteroaryl, 5 e. a heterocycle, f. an alkyl-C(O)-, the alkyl of which can be C 1

-C

6 , g. an aryl-C(O)-, h. a heteroaryl-C(O)-, 5 i. a heterocycle-C(O)-, j. a group represented by one of formulae (Ill) or (IV) 0 HN NH H H 0 s NO 2 (Ill) (IV) or k. an -0-C(O)- group or 10 1. an -NRa-C(0)- group in which Ra is as defined previously o If A with X together with the carbons to which they are attached represent a chain of formula (VI) R9 R10 R11 R12 A x Y R13 (VI) in which 15 e Y can represent a ketone (=0) group, an (=N-OH) oxime group or an alkyl (=N-O-alkyl) oxime group the alkyl group of which can be C 1

-C

6 ; * R 9 can represent a hydrogen atom, a C 1

-C

6 alkyl group, a halogen atom; or e R 9 and R 11 together can form an additional carbon-carbon bond between the carbon atoms to which they are attached, or a C 3

-C

6 cycloalkyl group; 20 e R 10 can represent a hydrogen atom, a halogen atom or an -ORa, -SRa, -CN, -NRaR group, -Ra and -Rb being as defined previously; 6 * R 1 1 can represent a hydrogen atom, or a C1-C6 alkyl, C3-C6 cycloalkyl, aryl group or a halogen atom; e R 12 can represent a hydrogen atom, or a C1-C6 alkyl group or a halogen atom or a -CN, -ORa, -SRa, -SERa, -C(0)-Ra 5 -C(0)ORa, -NRaRb, -OC(O)NRaRb group, -Ra and -Rb being as defined previously; or " R 11 and R 12 together with the carbon to which they are attached, can form a C3-C6 cycloakyl group; * R 1 3 can represent a hydrogen atom or a C1-C6 alkyl group, or a halogen atom 10 or a -CN, -CF 3 , -NO 2 , -ORa, -SRa, -SO 2 Ra, -NRaRb, -C(O)-Ra, -OC(O)Ra, -OC(O)NRaRb, -C(O)ORa, -C(O)NRaRb group or together with Z a carbon-carbon bond then " Z can represent a hydrogen atom, a hydroxyl group, a hydroxyamino group or 15 together with R 13 a carbon-carbon bond and " R 1 can represent a hydrogen atom or a -CH 3 , -CH 2 -CN,

-CH

2 -SRa, -CH 2 -SeRa group or also a group corresponding to the following formula (VII) or (VIll):

-CH

2 -W-R' (VII) or -C(O)-W-Rc (VIII) 20 in which + W can represent an oxygen atom or an -NRa group in which Ra is as defined previously or, a spacer arm constituted by an optionally substituted, linear or branched hydrocarbon chain comprising 2 to 20 carbon atoms and comprising moreover at least one heteroatom; 25 + RC being as defined previously; as well as: - its SYN, ANTI geometrical isomers, when they exist, - its optical isomers (enantiomers, diastereoisomers), when they exist, - its addition salts with a pharmaceutically acceptable acid or base, 30 - its hydrates and its solvates, 7 - its prodrugs, - or one of its esters. As a person skilled in the art understands, a certain number of compounds of formula (1) which comprise one or more hydroxyl groups, can be esterified. These 5 esters as well as their addition salts with pharmaceutically acceptable acids are not generally directly active in themselves but constitute prodrugs for the corresponding hydroxylated analogues. These esters, which are metabolized in the human organism, lead to active compounds. These esters are also the subject of the present invention. The esters introducing chemical functionalities such as sulphates, 10 phosphates, acids and basic chains which increase aqueous solubility and bioavailabilty can be mentioned. The esters of compounds bearing a basic function such as the analogues of dialkylglycine with alkyls with 1 to 4 carbon atoms and quite particularly dimethylglycine and diethylglycine and also methylpiperazine are preferred. The esters of fatty acids or the esters of a polyethyleneglycol chain which 15 increase the affinity for the lipophilic phases can be mentioned. Saturated fatty acid chains with 3 to 18 carbons are preferred. Moreover according to the present text, > the term "Cx-Cy alkyl refers to a linear or branched hydrocarbon radical, comprising x to y carbon atoms. Thus by way of example, the invention according 20 to the cases listed covers linear or branched hydrocarbon radicals, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, n hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl. The C-C 6 alkyl groups are preferred. The alkyl groups can optionally be substituted by an aryl group as defined hereafter, in which case an arylalkyl group is mentioned. 25 Examples of arylalkyl groups are in particular benzyl and phenethyl. Optionally, the alkyl groups can be substituted one or more times by one or more identical or different substituents, chosen independently from a halogen atom or a -CN, -CF 3 , -C(O)-Ra, -C(O)ORa, -C(O)NRaRb, -O-C(O)NRaRb, -NRaRb, -ORa, -SRb group, the R' and Rb groups can be as described previously. 30 > The term "Cx-Cy alkenyl" refers to a linear or branched or cyclic hydrocarbon radical, comprising one or more double bonds, having x to y carbon atoms. For example, the ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1- 8 pentenyl, 2-pentenyl, 3-methyl-3-butenyl, 1-hexenyl, 1-heptenyl, 1-octenyl radical can be mentioned. Optionally, the alkenyl groups can be substituted one or more times by one or more identical or different substituents chosen independently from a halogen atom or a -CN, -CF 3 , -C(O)-Ra, -C(O)ORa, -C(O)NRaRb, -O-C(O)NRaR, 5 -NRa Rb, -ORa, -SRb group, the R', Rb groups can be as described previously; > the term "Cx-Cy cycloalkyl " refers to a saturated or partially unsaturated cyclic hydrocarbon radical, having x to y carbon atoms. The cycloalkyl groups include in particular the substituents cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl. 10 Optionally, the cycloalkyl groups can be substituted one or more times by one or more identical or different substituents chosen independently from a halogen atom or a -CN, -CF 3 , -C(O)-Ra, -C(O)ORa, -C(O)NRaRb, -O-C(O)NRaRb, -NRaRb, -ORa SRa group, the Ra, Rb group can be as described previously; > the term "Cx-Cy alkynyl" refers to a linear, branched hydrocarbon radical 15 comprising at least one triple bond, having x to y carbon atoms. The alkynyl groups include in particular the ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2 butynyl, 1-pentynyl, 2-pentynyl, 1-heptynyl, 2-heptynyl, 1-octynyl, 2-octynyl substituents. Optionally, the alkynyl groups can be substituted one or more times by one or more identical or different substituents chosen independently from a 20 halogen atom or a -CN, -CF 3 , -C(O)-Ra, -C(O)ORa, -C(O)NRaRb, -O-C(O)NRaR b, -NRaRb, -ORa, -SRa group, the Ra, Rb groups can be as described previously; > the term "Cx-Cy aryl" refers to an aromatic hydrocarbon radical, having x to y carbon atoms. Preferentially according to the invention the aromatic hydrocarbon radicals having 6 carbon atoms are preferred. The aryl groups 25 include in particular the phenyl, naphthyl and bi-phenyl radicals. Optionally, the aryl groups can be substituted one or more times by one or more identical or different substituents chosen independently from a halogen atom or an alkyl, -CN,

-CF

3 , -N 3 , -NO 2 , -C(O)ORa, -C(O)-Ra, -C(O)NRaR, -O-C(O)NRaR, -NRaRb, -ORa -SRa group, the Ra, Rb groups can be as described previously; 30 > the term "Cx-Cy heterocycle" refers to a mono- or poly-cyclic, saturated, unsaturated or aromatic, optionally substituted radical and can comprise x to y carbon atoms and comprising one or more heteroatoms. Preferably, the 9 heteroatoms are chosen from oxygen, sulphur and nitrogen. Examples of heterocycles are the furyl, thienyl, pyrrole, imidazole, isothiazole, thiazole, isoxazole, oxazole, pyridine, pyrazine, pyrimidine, pyridazine, indole, isoindole, indazole, quinoline, isoquinoline, phthalazine, quinazoline, pyrrolidine, 5 imidazolidine, pyrrazolidine, piperidine, piperazine, morpholine, thiazolidine or phthalimide, benzimidazole radicals. Optionally, the heterocyle groups can be substituted one or more times by one or more identical or different substituents chosen independently from a halogen atom or an alkyl, -CN, -CF 3 , -N 3 , -NO 2 , C(O)ORa, -C(O)-Ra, -C(O)NRaRb, -O-C(O)NRaR, -NRaRb, -ORa, -SRa group, the 10 Ra, Rb groups can be as described previously; > the term "halogen" refers to a chlorine, bromine, fluorine and iodine atom. Preferentially according to the invention, the halogen is a fluorine atom. The compounds of formula (1) are known and are particularly described in the international applications WO 2006/027454 of 13 March 2006, WO 2007/101925 15 of 13 September 2007, WO 2009/044011 of 09 September 2009, WO 2010/076418 of 08 July 2010, WO 2004/082581 of 30 September 2004, WO 2007/080270 of 19 July 2007, W02007/118967 of 25 October 2007, WO 008/142237 of 27 November 2008, WO 2009/044010 of 09 April 2009, WO 2009/092892 of 30 July 2009 and WO 2010/012904 of 4 February 2010. 20 It is well known (see the international applications cited previously) that the cholesterol derivatives, such as particularly for example those of formula (1), particularly 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime and cholest-4-en-3-one oxime can be used as medicaments and particularly have remarkable cytoprotective properties, particularly neuroprotective, card ioprotective and/or 25 hepatoprotective. But these compounds are very lipophilic and very insoluble in aqueous medium compatible with an administration for example a parenteral administration, which makes their introduction into compositions, particularly pharmaceutical compositions, particularly difficult or even impossible. It is therefore difficult to 30 obtain such compositions that are chemically and physically stable. In the prior art, the development of compositions comprising active ingredients having a low aqueous solubility, traditionally focused on the use of surfactants 10 which allow the formation of emulsions, colloids such as micelles or liposomes which solubilize the medicament and increase its solubility in aqueous medium. Nevertheless, these emulsions and in particular the micellar suspensions are not physically or even chemically stable. For example if the composition comes into 5 contact with blood or plasma, the solubilizing system can thus lose these properties, retaining only the active ingredient. Moreover, the use of surfactant (cremophore, tween, etc.) and the formation of particulate structures give rise to the activation of the complement system of higher organisms and the initiation of reactogenicity reactions which can be fatal. 10 These problems of solubility and reactogenicity become all the more difficult when it is sought to obtain a composition having a high concentration of active ingredient, allowing inter alia the administration of a volume of composition as small as possible or in a shorter time to be envisaged. The invention aims inter alia to overcome these problems and difficulties. 15 The applicant has, surprisingly, discovered that the cholesterol derivatives, particularly those described in the international applications cited previously, advantageously those corresponding to formula (1), have a very good affinity for liposomes, which moreover allows the preparation of compositions, particularly pharmaceutical compositions, which are physically and chemically stable for 20 several months and concentrated with active ingredient. In addition these compositions when in contact with a biological medium such as blood are chemically stable and the active ingredient retains all its properties. Moreover, these compositions once administered surprisingly have a very low reactogenicity. 25 Moreover, the liposomes, according to the invention, have a prolonged remanence in biological liquids, particularly in blood. The invention resides in the fact that the liposomes according to the invention allow a much more significant solubilization of the compounds of formula (1) while having a very low reactogenicity. Thus the compounds of formula (1), when 30 included in the liposomes according to the invention, can be formulated in pharmaceutical compositions in contact with aqueous media much more easily than if they were presented in another form.

11 The liposomes consist of at least one lipid bilayer membrane surrounding an aqueous internal compartment. They are known as an effective system of formulations for transporting therapeutic agents, drugs, or active ingredients within the aqueous space present inside the vesicle (agents soluble in aqueous medium) 5 or included in the lipid bilayer (agents insoluble in aqueous medium). They can be characterized by the type of membrane and their size. The unilamellar vesicles are constituted by a single bilayer membrane. The multilamellar vesicles (MLV: multilamellar vesicle) have at least two lipid bilayer membranes defining several aqueous closed compartments. The membranes are organized in a concentric 10 fashion so that the different membranes are separated by an aqueous compartment. The small unilamellar vesicles (SUV: small unilamellar vesicle) can have a diameter generally comprised between 20 and 100 nm. The large unilamellar vesicles (LUV: large unilamellar vesicle) and the multilamellar vesicles (MLV: 15 multilamellar vesicle) can have a diameter generally greater than 100 nm. As regards the liposomes, it is possible to refer to the work of Gregory Gregoriadis (Liposome Technology: Liposome Preparation And Related Techniques, 3rd edition, 2007) for their description, manufacture and applications. But one of the major drawbacks with the development of the liposomal 20 formulations, on the experimental scale for the pharmaceutical market, is their chemical and physical instability and their relative stability during production and storage. Therefore, since Bangham in 1965 (Bangham AD., Standish MM., Watkins JC. (1965). J. Mol. Biol. 13, p. 238-252) who was the first to manufacture liposomes, 25 numerous improvements have been made in particular for increasing their steric stabilization and for increasing their residence time in the vascular system which in particular makes it possible to limit injections or even to envisage targeted therapies or delayed forms. Among these improvements "PEGylation" can be mentioned as an example 30 which consists of the introduction of PEG (Poly Ethylene Glycol) chains often grafted on phospholipids or cholesterol.

12 The introduction of sterols such as cholesterol into the lipid bilayer allows the stabilization of the liposomes to be improved. The patent US 6,143,321 describes liposomes the active ingredient of which is entrapped or adsorbed in the lipid bilayer using a surfactant. 5 An improvement in the physical and chemical stability of the liposomes according to the invention represents one of the great advantages of the invention as it makes it possible to envisage the preparation of low-volume compositions comprising a large quantity of active ingredient, which would make it possible to envisage, particularly in the case of pharmaceutical compositions, delivering to 10 patients only small doses or even a single dose of medicaments, which can make it possible to limit or even eliminate hypersensibilization (reactogenicity) problems that could cause the administration of several doses of medicament solubilized using liposomes. In addition, as the active ingredient can be comprised in the liposomes 15 according to the invention, it is sufficient in itself to confer an improved stability on the liposomes without adding extra sterol. Another major drawback in the pharmaceutical development of liposomal formulation in particular by parenteral route is the increased risk of reactogenicity linked to the activation of the complement (Szebeni J., Toxicology, 2005, 216, 20 106). These reactions are even more frequent or intense when the administration of the solution is carried out at a high rate such as with a bolus. A weakly reactogenic pharmaceutical composition would allow a parenteral administration by bolus and not by infusion of a massive quantity of active ingredient. Moreover this formulation would allow a rapid administration by bolus and preferentially 25 manually. By "weakly reactogenic pharmaceutical composition" is meant in the present text that the group having received the composition induces a level of reactogenicity similar to that induced in the control group of the trial having only received the buffer of this composition. 30 An administration by bolus corresponds to the administration of a quantity necessary in order to obtain the expected concentration in the blood, the lymph, the cerebrospinal fluid or any targeted biological site for therapeutic or diagnostic 13 reasons in a short period of time comprised between 1 second and 10 minutes, preferentially in less than 5 minutes, very preferentially in less than 2 minutes. A weakly reactogenic composition increases the level of the complement by a maximum of three times with respect to the basal level measured in the serum of 5 the individual. The level of terminal C complex (SC5b-9) can be measured by an immunological test kit such as that from the Quibel Corporation "SC5b-9 plus Elisa" Kits. It is one of the purposes of the invention to provide weakly reactogenic liposomes, which can be used alone or in pharmaceutical compositions. 10 Thus the properties of the liposomes according to the invention make it possible to prepare compositions, preferentially pharmaceutical compositions, having the following advantages: - physical and chemical stability for at least 12 months in storage phase at 25*C; 15 - possible use for an administration by bolus by parenteral route; - very low reactogenicity; - process which can be adapted to industrial production; - integration with other active ingredients, with the possible option of a massive administration by bolus. 20 Thus a first subject of the invention is liposomes comprising at least one of the compounds corresponding to formula (1) or a mixture of compounds corresponding to formula (1). A subject of the invention is also a composition, particularly a pharmaceutical composition comprising liposomes comprising at least one of the compounds 25 corresponding to formula (1) or a mixture of compounds corresponding to formula (1). A person skilled in the art has a good knowledge of liposomes and the techniques not only for preparing them but also for introducing an active agent into them. The invention relates to any known type of liposome providing it is used to 30 encapsulate an active agent, particularly a compound of formula (1).

14 According to a particular form of the invention, the final liposomal solution can comprise at least: - a compound of formula (1); - a phospholipid or a mixture of phospholipids; 5 - an agent that stabilizes the pH; - and optionally a cryoprotective agent. Advantageously the compound of formula (1) comprised in the liposomes according to the invention can be chosen from cholest-4-en-3-one oxime, 10 cholest-4,24-dien-3-one oxime, cholestan-3-one oxime, 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, 1,4-cholestadien-3-one oxime, 2-methyl-cholest-4-en-3-one oxime, 15 2a-fluoro-cholest-4-en-3-one oxime, 4-methoxy-cholest-4-en-3-one oxime, 4-fluoro-cholest-4-en-3-one oxime, 6p-fluoro-cholest-4-en-3-one oxime, 19-hydroxy-cholest-4-en-3-one oxime, 20 19-biotinyloxy-cholest-4-en-3-one oxime, 25-((N-(+)-biotinoyl-N-methyl) amino)-27-norcholest-4-en-3-one oxime, 25-[methyl(7-nitro-2.1,3-benzoxadiazol-4-yl)amino]-27-norcholest-4-en-3-one oxime, 2,2-difluoro-cholest-4-en-3-one oxime, 25 2,6-difluoro-cholest-4-en-3-one oxime, cholest-4-en-3,6-dione 3-oxime, cholest-4,21-dien-3,6-dione 3-oxime, 24-ethyl-cholest-4-en-3,6-dione 3-oxime, 24-ethyl-cholest-4,21-dien-3,6-dione 3-oxime, 15 24-methyl-cholest-4,21-dien-3,6-dione 3-oxime, 3-[methyl(7-nitro-2.1,3-benzoxadiazol-4-yl)amino]-3,5-seco-4-norcholestan-5 one oxime, 2-hydroxy-2,5-seco-3,4-dinor-cholestan-5-one oxime, 5 3-[(N-(+)-biotinoyl-N-methyl)amino]-3,5-seco-4-norcholestan-5-one oxime, 25-fluoro-3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, 3,5-seco-4-norcholestan-5-one oxime, 3-hydroxy-3,5-seco-4-norcholest-24-en-5-one oxime, 3-hydroxy-3,5-seco-3-trifluoromethyl-4-norcholestan-5-one oxime, 10 24p-ethyl-3-hydroxy-3,5-seco-4-norcholest-22-en-5-one oxime, 3-hydroxy-3-methyl-3,5-seco-4-norcholestan-5-one oxime, 3,3-dimethyl-3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, cholest-4-en-3,6-dione dioxime, 15 cholest-4,24-dien-3,6-dione dioxime, 24-methyl-cholest-4,21-dien-3,6-dione dioxime, 24-ethyl-cholest-4-en-3,6-dione dioxime, 24-ethyl-cholest-4,21-dien-3,6-dione dioxime, cholest-4-en-3-one, 20 cholest-4,24-dien-3-one, 3-hydroxy-3,5-seco-4-norcholestan-5-one, 3-hydroxy-3-methyl-3,5-seco-4-norcholestan-5-one, 3,3-dimethyl-3-hydroxy-3,5-seco-4-norcholestan-5-one, 3-hydroxy-3,5-seco-4-norcholestan-5-one methyloxime. 25 Very preferentially the compound of formula (I) comprised in the liposomes according to the invention can be chosen from cholest-4-en-3-one oxime, cholestan-3-one oxime, cholest-1,4-dien-3-one oxime, cholest-4,24-dien-3-one oxime, 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, 4-fluoro-cholest-4-en-3-one oxime, 3-hydroxy-3-methyl-3,5-seco-4-norcholestan-5-one oxime, 3,3-methyl-3- 16 hydroxy-3,5-seco-4-norcholestan-5-one oxime, very preferentially cholest-4-en-3 one oxime and 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime. Of course, the invention also relates to the liposomes for which a mixture of at least two compounds of formula (1) can be used for their constitution. 5 According to the invention the compound of formula (1) can be present in the liposome at physiologically effective doses. According to the invention, the compound of formula (1) can be present, in the final liposomal solution, in a quantity ranging from 0.1 to 200 mg/mL, preferentially below 1 mg/mL, also more preferentially below 5 mg/mL, very preferentially below 10 mg/mL). 10 By "final liposomal solution" is meant the solution of liposomes obtained after implementation of the process making it possible to obtain the liposomes according to the invention. According to the invention, the phospholipids used can be phospholipids of natural, vegetable or animal, or synthetic origin, yet more advantageously natural 15 phospholipids. Said phospholipids can be chosen from phosphoacylglycerols (better known under the name of glycerophospholipids), inositophosphatides, phosphosphingolipids and phosphonosphingolipids or also phosphosaccharolipids. Preferentially the phospholipids can be chosen from phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, 20 linoleylpalmitoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, oleoylpalmitoylphophatidylcholine, DHAstearoylphosphatidyl choline, DHA-rich phosphatidylcholine of avian origin, phosphatidylinositol, DHA phosphatidylethanolamine, phosphatidylserine, sphingomyelin, a mixture of phospholipids of avian origin close to the composition of human milk, a mixture of 25 phospholipids of soya origin close to the composition of human milk, lysophosphatidic palmitic or oleic acids, egg lysophosphatidylcholine containing palmitic and stearic acids at more than 90%, soya lysophosphatidylcholine, lysophosphatidylinositol, lysophosphatidylethanolamine, lysophosphatidylserine, a mixture of egg phospholipids containing phosphatidylcholine, 30 phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and sphingomyelin, 1,2-dioleolyl-sn-glycero-3-phosphocholine (DOPC), 1,2 Dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC), 1,2- 17 dimyristoylphosphatidylglycerol (DMPG), egg L-a-phosphatidylcholine, soya L-a phosphatidylcholine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N [methoxy (polyethylene glycol)-2000] (DSPE-PEG). Preferably, according to the invention phospholipids of natural origin are used 5 and particularly egg or soya phospholipids, preferably egg phospholipids. Of course, the invention also relates to liposomes for which a mixture of at least two phospholipids can be used for their constitution. According to the invention the phospholipids can be present in the final liposomal solution, in a quantity ranging from 10 to 300 mg/mL, preferentially 10 between 20 and 200 mg/mL, very preferentially between 50 and 150 mg/mL). A person skilled in the art will without difficulty establish the ratio of the concentration of the compound of formula (1) to that of the phospholipid to be used in order to obtain the liposomes according to the invention, particularly with reference to Gregory Gregoriadis (Liposome Technology: Liposome Preparation 15 And Related Techniques, 3rd edition, 2007). By way of example, but without being limitative, it can be indicated that the ratio of the concentration of the compound of formula (1) to that of the phospholipid can be comprised in the final liposomal solution between 1/100 and 70/100, preferentially between 10/100 and 50/100. 20 According to the invention, the pH stabilizing agent can be a phosphate, benzoate, citrate, glutamate, lactate, ascorbate, tartrate, succinate, adipate, glycinate, malate, triethanolaminate, diethanolaminate, tromethaminate buffer. Preferably, according to the invention a phosphate buffer is used. According to the invention the pH stabilizing agent can allow the pH of the 25 liposomal formulation to vary between 3 and 11, preferentially between 4 and 9. A person skilled in the art will without difficulty adjust the quantity of stabilizing agent to be introduced as a function on the one hand of the final pH of the liposomal formulation that it is intended to obtain and as a function of the buffer chosen and its physical properties. 30 Similarly according to the invention, the cryoprotective agent can be for example glycerol, sucrose, dextrose, trehalose, glucose, maltose, mannose, lactose, mannitol, sorbitol, glycine, polyvinylpyrrolidone (PVP), polyvinylalcohol 18 (PVA), gelatin, alanine, lysine, polyethylene glycol, dextran, aerosil, fructose, hydroxypropyl-p-cyclodextrin. Preferentially according to the invention, glycerol, sucrose, dextrose are used . According to the invention the cryoprotective agent can be present in the 5 liposomal formulation in a quantity ranging from 0.01 to 30% in solution, preferentially between 0.1 and 20%, very preferentially between 1 and 10%. According to the invention the compound of formula (1) can be combined with other active therapeutic agents. Thus, it is possible to combine this active ingredient with another complementary or secondary ingredient incorporated either 10 in said lipid layer of the liposome, or in the aqueous phase depending on their solubility. The liposome according to the present invention can be used alone or in a composition in animals or humans, particularly mammals, more precisely in humans. They can be for cosmetic, pharmaceutical or veterinary use. 15 Thus a subject of the invention is also a composition comprising at least one liposome comprising at least one compound of formula (1). It is known that the composition of the invention can also contain the usual adjuvants in the fields considered depending on the administration method, such as for example preservatives, antioxidants, pigments and colouring materials, 20 thickeners, fragrances, sweeteners, agents stabilizing the active particles. The quantities of these different adjuvants are those used in a standard fashion in the fields considered, and are for example from 0.0001 % to 10 % of the total weight of the composition. These adjuvants are introduced into the aqueous or lipophilic phase. 25 According to the invention, the liposome or the composition comprising at least one liposome, can be administered by enteral, parenteral or topical route, preferentially by parenteral route. By parenteral route, the administration can be carried out by intravenous or intraarterial or intralymphatic, direct (syringe) or indirect route (by a perfusion or an angioplasty catheter). 30 In the case of a vein this can be superficial, usually in the arm (peripheral venous route) or deep (central venous route), most often in the neck (jugular vein) or under the clavicle (sub-clavian vein), by sub-cutaneous route, under the skin, 19 frequently in the stomach or thighs, by intradermal route, directly into the dermis, by intramuscular route, directly into a muscle or also by pulmonary route by inhalation. Preferentially, the administration can be carried out by intravenous route. 5 A subject of the invention is also the use of the compounds of formula (I) for preparing liposomes, advantageously stable liposomes. A subject of the invention is also the use of the liposomes according to the invention for preparing a composition, advantageously a weakly reactogenic, cosmetic, pharmaceutical, or veterinary composition. 10 Moreover, a subject of the invention is the use of the liposomes according to the invention for the transport of active ingredients other than the compounds of formula (1). Other features and properties of the invention will become apparent on reading the following examples which illustrate the invention without thereby limiting it. 15 Abbreviations used: EPC: Egg L-a-phosphatidylcholine SPC: Soya L-a-phosphatidylcholine DOPC: 1,2-dioleoyl-sn-glycero-3-phosphocholine DMPG: 1.2 -dimyristoylphosphatidylglycerol 20 DSPE-PEG: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycol)-2000] PBS: Phosphate buffer saline Eth: Ethanol terBut: tert-Butanol 25 List of the compounds of formula (1) tested: N* Chemical name 1 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime 2 3,3-dimethyl-3-hydroxy-3,5-seco-4-norcholestan-5-one oxime 3 cholest-4-en-3-one oxime 4 cholest-4,24-dien-3-one oxime 20 5 4-fluoro-cholest-4-en-3-one oxime 6 3,5-seco-4-norcholestan-5-one oxime 7 cholest-4-en-3-one 8 3-hydroxy-3,5-seco-4-norcholestan-5-one 9 cholest-4,24-dien-3-one Example 1: comparison of the solubility of the derivatives of formula (1) The maximum solubility of the compounds of formula (1) was tested in water and different solvents: Compound N* 1 3 4 6 Solvent Water 0.001 0.0001 0.0001 0.0001 Soya oil 6 30 (2-hydroxypropyl)-beta-cyclodextrin, 0.7 30% PBS buffer Intralipid@10 1 0.34 0.4 Intralipid@10/SolutolHS S5/ethanol 5 92/3/5 Liposome based on egg 30 12.4 14.5 8.2 phospholipids Concentration in mg/mL 5 Intralipid@: injectable lipid emulsion, generally used in parenteral nutrition, sold by the laboratory Fresenius Kabi France Solutol HS15@: non ionic solubilizing agent sold by the company BASF. CONCLUSION: The liposomal formulation allows a solubility of the compounds of formula (1) in an aqueous medium of greater than or equal to 8 mg/mL and at a 10 concentration equal to or greater than a medium which is highly lipophilic but not miscible in an aqueous biological medium. Example 2: preparation of suspensions of liposomes containing the derivatives of formula (1) A solution of lipids at 250 mg/mL is prepared by dissolving defined quantities of 15 EPC and the compound of formula (1) in tert-butanol or a mixture of tert-butanol and absolute ethanol (v/v). This lipid solution is mixed at a certain temperature 21 with aqueous buffer in order to obtain a final concentration of solvent of 15 to 17% and of product of formula (1) of 25 or 50 mg/mL. The large multilamellar vesicles are then extruded at a fixed temperature through 3 polycarbonate filters by 10 consecutive passes. The non encapsulated 5 compound of formula (1) is removed on the filters. The solvent is eliminated by filtration on membrane by washing with buffer in order to achieve a percentage of final solvent of the order of 0.4%. A B C D E F G H 1 Eth/terBut 25 25 50/200 8.1 15-20 81.4 2 Eth/terBut 25 25 50/200 9.2 15-20 79.5 3 Eth/terBut 40 45 50/200 12.4 15-20 88.9 4 Eth/terBut 80 45 50/200 14.5 15-20 80.7 5 terBut 80 60 25/100 3.6 30-45 79.1 6 Eth/terBut 80 55 50/200 8.2 25-35 84.7 7 Eth/terBut 25 25 50/200 12.1 15-20 8 Eth/terBut 25 25 50/200 11.4 15-20 9 Eth/terBut 25 25 50/200 10.1 15-20 A: Compound B: Solvent(s)(v/v) 10 C: Solubilization temperature in 'C D: Extrusion temperature in *C E: Initial concentration in mg/mL of buffer [Compound/EPC] F: Concentration of compound in mg/mL in the liposomes G: Extrusion pressure (Bars) 15 H: Particle size (nm) CONCLUSION: The solutions of liposomes with the compounds were produced with concentrations between 3 and 15 mg/mL. The size of the vesicles is very 22 homogenous from one compound to the other, of the order of 80 nm corresponding to small unilamellar vesicles. Example 3: preparation of suspensions of liposomes containing 3 hydroxy-3,5-seco-4-norcholestan-5-one oxime (compound 1) 5 A solution of lipids at 250 mg/mL is prepared by dissolving defined quantities of phospholipids and compound 1 in a mixture of tert-butanol and absolute ethanol (v/v). This lipid solution is mixed at ambient temperature with aqueous buffer in order to obtain a final concentration of solvent of 20% and of total lipids of 50 mg/mL. 10 The large multilamellar vesicles are then extruded at ambient temperature through 3 to 5 polycarbonate filters. 5 to 10 passes are necessary to generate the small unilamellar vesicles with a size between 70 and 90 nm. The solvent and non encapsulated 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime are removed by diafiltration using 10 volumes of washing buffer. An 15 ultrafiltration is then carried out in order to concentrate the formulation to the final concentration. A B C D E F 1 EPC 10 mM Phosphate 81.40 31.30 73.00 300 mM Sucrose 2 DOPC 10 mM Phosphate 51.50 22.06 212.00 DSPE-PEG (96/4) 300 mM Sucrose 3 EPC 10 mM Phosphate 53.20 25.30 70.80 145 mMNaCI 4 SPC-DMPG (96/4) 10 mM Phosphate 63.30 27.80 70.70 SPC-DPG (6/4) 145 mMNaCI 5 DOPC-DMPG 10 mM Phosphate 71.70 29.30 95.70 (96/4) 145 mMNaCI 6 EPC 10 mM Phosphate 63.40 26.20 57.40 300 mM sucrose 10 mM Phosphate 7 EPC 300 mM sucrose 46.30 21.88 145 mMNaCI 8 EPC 10 mM Phosphate 46.30 22.00 5% dextrose 23 145 mMNaCI 10 mM Phosphate 9 EPC 5% glycerol 46.30 22.00 145 mMNaCI 10 EPC 10 mM Phosphate 66.50 27.70 69.20 5% glycerol 11 EPC 10 mM Phosphate 76.7 19.6 67.0 5% glycerol A: Composition (compound 1 + phospholipids of column B) B: Phospholipids used for preparing the liposomes C: Buffer D: Concentration of phospholipids in mg/mL 5 E: Final concentration of compound 1 in mg/mL F: Size of the vesicles in nm CONCLUSION: The different liposomal compositions with concentrations around 20 to 35 mg/mL have small unilamellar vesicles of 50 to 100 nm, with the exception of composition 2. A low percentage of DSPE-PEG appears to form 10 large unilamellar or multilamellar vesicles of 220 nm in the same final concentration range in compound 1. Example 4: In vitro evaluation of the reactogenicity The in vitro evaluation of the reactogenicity is carried out by measuring the production of SC5b-9 complex after incubation in human serum. The levels of 15 SC5b-9 are measured with an ELISA kit according to the method described (QuidelCorporation; SC5b-9 More EIA kit, Ref. A029). The solutions tested are diluted with buffer in order to achieve a concentration of compound of formula (1) of 20 mg/mL. 1 volume of liposomal solution prepared in Example 3 is added to 3 volumes of serum in order to achieve a tested final 20 concentration of 5 mg/mL. The mixture is stirred vigorously and incubated for 45 minutes at 37*C. The reaction is stopped with the "stop" solution from the Elisa kit. The levels of production of the SC5b-9 complex in the sera were measured with, as control: 24 e the average of the concentrations obtained over all the sera alone (negative control) (Cl), * the buffers (T1, T2) e a positive control (Zymosan-A@, polysaccharide of yeasts sold by SIGMA 5 ALDRICH) * a reference compound (AmBisome@) (positive liposomal control). N* Solution A B C D Serum 0.7 Zymosan@ 170 Ambisome@ 30.7 T1 10 mM Phosphate 2.3 300 mM Sucrose T2 10 mM Phosphate 5.3 145 mMNaCI 1 EPC 10 mM Phosphate 31.30 8.4 300 mM Sucrose DOPC10m Phsht 2 DSPE-PEG 30 mM SuPhos e 22.06 9.7 (96/4) 3 EPC 10 mM Phosphate 25.30 7.8 145 mMNaCI 4 SPC-DMPG 10 mM Phosphate 27.80 7.6 (96/4) 145 mMNaCl 5 DOPC- 10 mM Phosphate 29.30 7.9 DMPG (96/4) 145 mMNaCI 10 mM Phosphate 7 EPC 300 mM sucrose 21.88 11.7 145 mMNaCI 10 mM Phosphate 8 EPC 5% dextrose 22.00 12.3 145 mMNaCI 10 mM Phosphate 9 EPC 5% glycerol 22.00 5.8 145 mMNaCI 10 EPC 10 mM Phosphate 27.70 2.1 5% glycerol

N

0 : number of the solution tested. A: Phospholipids B: Buffer 10 C: Concentration of compound mg/mL D: Concentration of SC5b-9(pg/mL) 25 CONCLUSION: The level of SC5b-9 obtained in the serum with the positive liposomal control Ambisome@ is 3 to 15 times greater than the levels obtained with the liposomal solutions prepared in Example 3. These induce the formation of SC5b-9 at low levels, similar to those obtained with the buffer or the serum alone. 5 The liposomal solutions prepared are therefore weakly reactogenic. Example 5: Evaluation of the reactogenicity of the liposomal formulation in humans. A Phase 1 clinical study was carried out with the liposomal formulation No. 11 exemplified in Example 3 of 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime. The 10 formulation was administered to 54 healthy volunteers by intravenous route in several doses and flow rates according to the protocol filed with AFSSAPS and with the "Comite de protection des personnes" (Committee on the protection of individuals). This formulation showed a very good tolerance up to the maximum dose tested of 13 mg/kg at 10 mL/min and with a maximum flow rate of 35 mL/min 15 for the dose of 10 mg/kg as well as a good demonstration of the product of formula (). CONCLUSION: No significant clinical sign relating to the activation of the complement was observed in humans in this study. 20 Example 6: Evaluation of the stability of the liposomal formulation The solution of liposome No. 11 exemplified in Example 3 of 3-hydroxy-3,5 seco-4-norcholestan-5-one oxime was subjected to stability testing at +5 and +25*C and analyzed after 3, 6, 12 months.

26 Parameters TO 3 months 6 months 12 months Translucent Translucent Translucent Translucent Appearance solution solution solution solution Crystallization Absence of Absence of Absence of Absence of crystals crystals crystals crystals Size of the vesicles (nm) 67.0 68.3 68.9 70.7 Concentration in mg/mL 19.6 19.2 19.2 18.4 Impurities (% of area) 2.38 2.29 2.52 2.25 Conclusion: The parameters measured are stable for at least a year at 25*C. The liposomal 5 solution can be stored for at least 1 year at 25 0

C

Claims (19)

1. 2. The liposome according to claim 1, wherein the compound of formula I is chosen from " 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, " 3-[methyl(7-nitro-2.1,3-benzoxadiazol-4-yl)amino]-3,5-seco-4-norcholestan-5 one oxime, " 2-hydroxy-2,5-seco-3,4-dinor-cholestan-5-one oxime, " 3-[(N-(+)-biotinoyl-N-methyl)amino]-3,5-seco-4-norcholestan-5-one oxime, " 25-fluoro-3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, " 3,5-seco-4-norcholestan-5-one oxime, " 3-hydroxy-3,5-seco-4-norcholest-24-en-5-one oxime, " 3-hydroxy-3,5-seco-3-trifluoromethyl-4-norcholestan-5-one oxime, " 24p-ethyl-3-hydroxy-3,5-seco-4-norcholest-22-en-5-one oxime, " 3-hydroxy-3-methyl-3,5-seco-4-norcholestan-5-one oxime, " 3,3-dimethyl-3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, " 3-hydroxy-3,5-seco-4-norcholestan-5-one oxime, " 3-hydroxy-3,5-seco-4-norcholestan-5-one, " 3-hydroxy-3-methyl-3,5-seco-4-norcholestan-5-one, " 3,3-dimethyl-3-hydroxy-3,5-seco-4-norcholestan-5-one, and " 3-hydroxy-3,5-seco-4-norcholestan-5-one methyloxime. 32
2. 3. The liposome according to claim 1 or claim 2, wherein the compound of formula I is present in the final liposomal solution in a quantity ranging from 0.1 to 200 mg/mL.
3. 4. The liposome according to claim 3, wherein the compound of formula I is present in the final liposomal solution in a quantity below 10 mg/mL.
4. 5. The liposome according to claim 3 or claim 4, wherein the compound of formula I is present in the final liposomal solution in a quantity below 5 mg/mL or below 1 mg/mL.
5. 6. The liposome according to any one of claims 1 to 5, characterized in that it comprises at least one compound of formula I or a mixture of compounds of formula I, a phospholipid or a mixture of phospholipids and a pH stabilizing agent.
6. 7. The liposome according to claim 6, characterized in that it comprises moreover a cryoprotective agent.
7. 8. The liposome according to claim 7, wherein the cryoprotective agent is chosen from glycerol, sucrose, dextrose, trehalose, glucose, maltose, mannose, lactose, mannitol, sorbitol, glycine, polyvinylpyrrolidone (PVP), polyvinylalcool (PVA), gelatin, alanine, lysine, polyethylene glycol, dextran, aerosil, fructose, hydroxypropyl-p-cyclodextrin, preferentially from glycerol, sucrose or dextrose.
8. 9. The liposome according to claim 7 or claim 8, wherein the cryoprotective agent is present in the liposomal formulation in a quantity ranging from 0.01 to 30% in solution.
9. 10. The liposome according to any one of claims 7 to 9, wherein the cryoprotective agent is present in the liposomal formulation in a quantity ranging from between 0.1 and 20%, preferentially between 1 and 10%. 33
10. 11. The liposome according to any one of claims 6 to 10, wherein the phospholipid is chosen from phosphoacylglycerols (better known under the name of glycerophospholipids), inositophosphatides, phosphosphingolipids and phosphonosphingolipids or also phosphosaccharolipids.
11. 12. The liposome according to claim 11, wherein the phospholipid is chosen from phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, linoleylpalmitoylphosphatidylcholine, palmitoyloleoylphosphatidylcholine, oleoylpalmitoylphophatidylcholine, DHAstearoylphosphatidyl choline, DHA-rich phosphatidylcholine of avian origin, phosphatidylinositol, DHA-phosphatidylethanolamine, phosphatidylserine, sphingomyelin, a mixture of phospholipids of avian origin close to the composition of human milk, a mixture of phospholipids of soya origin close to the composition of human milk, lysophosphatidic palmitic or oleic acids, egg lysophosphatidylcholine containing palmitic and stearic acids at more than 90%, soya lysophosphatidylcholine, lysophosphatidylinositol, lysophosphatidylethanolamine, lysophosphatidylserine, a mixture of egg phospholipids containing phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol and sphingomyelin, 1,2-dioleolyl-sn-glycero-3-phosphocholine (DOPC), 1,2-Dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC), 1,2 dimyristoylphosphatidylglycerol (DMPG), egg L-a-phosphatidylcholine, soya L-a phosphatidylcholine, and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N [methoxy (polyethylene glycol)-2000] (DSPE-PEG).
12. 13. The liposome according to any one of claims 6 to 12, wherein the phospholipid is chosen from egg or soya phospholipids, preferably egg phospholipids.
13. 14. The liposome according to any one of claims 6 to 13, wherein the phospholipid is present in the final liposomal solution in a quantity ranging from 10 to 300 mg/mL.
14. 15. The liposome according to claim 14, wherein the phospholipid is present in the final liposomal solution in a quantity ranging from between 20 and 200 mg/mL, preferentially between 50 and 150 mg/mL. 34
15. 16. The liposome according to any one of claims 6 to 15, wherein the pH stabilizing agent is a phosphate, benzoate, citrate, glutamate, lactate, ascorbate, tartrate, succinate, adipate, glycinate, malate, triethanolaminate, diethanolaminate, or tromethaminate buffer, advantageously a phosphate buffer.
16. 17. A composition comprising at least one liposome comprising at least one compound of formula (I) as described in any one of claims 1 to 16.
17. 18. Use of at least one compound of formula (I) as defined in claim 1 for preparing liposomes.
18. 19. Use of the liposomes according to any one of claims 1 to 16 for preparing a composition, advantageously a weakly reactogenic, cosmetic, pharmaceutical, or veterinary composition.
19. 20. Use of the liposomes according to any one of claims 1 to 16 for the transport of active ingredients other than the compounds of formula (I). Trophos Patent Attorneys for the Applicant/Nominated Person SPRUSON & FERGUSON