Composition Intended to Vectorise an Anti-Cancer Agent

Abstract

A composition in the form of a water-in-oil emulsion comprising: from 20% to 40% (v/v) of aqueous phase, in the form of droplets, comprising an anti-cancer agent and a densifying agent chosen from the complexes of nonionic macrocyclic chelate with a paramagnetic metal, from 60% to 80% (v/v) of lipid phase comprising an iodized oil and at least one surfactant of formula (I) in a proportion, by weight of surfactant relative to the total volume of the composition, of 0.3% to 5%, formula (I) of said surfactant being the following:

##STR00001##

in which: s is 0 or 1, m is an integer from 2 to 30, R.sub.1 represents a group of formula (II)

##STR00002## in which n is an integer from 4 to 10, o is an integer from 1 to 4, p is an integer from 3 to 7, q is an integer from 2 to 10, and r is 0 or 1, R.sub.2 represents a hydrogen atom or is identical to R.sub.1, and each R.sub.3 independently represents a hydrogen atom or is identical to R.sub.1.

Claims

1-15. (canceled)

16. A composition in the form of a water-in-oil emulsion comprising: from 20% to 40% (v/v) of aqueous phase, in the form of droplets, comprising an anti-cancer agent and a densifying agent selected from complexes of a nonionic macrocyclic chelate with a paramagnetic metal; from 60% to 80% (v/v) of lipid phase comprising an iodized oil and at least one surfactant of formula (I) in a proportion, by weight of surfactant relative to the total volume of the composition, of 0.3% to 5%, wherein formula (I) of said surfactant is the following: ##STR00012## wherein: s is 0 or 1; m is an integer from 2 to 30; R.sub.1 represents a group of formula (II) ##STR00013## wherein n is an integer from 4 to 10, o is an integer from 1 to 4, p is an integer from 3 to 7, q is an integer from 2 to 10, and r is 0 or 1; R.sub.2 represents a hydrogen atom or an R.sub.1; and each R.sub.3 independently represents a hydrogen atom or an R.sub.1.

17. The composition of claim 16, wherein each R.sub.3 is a hydrogen atom.

18. The composition of claim 16, wherein the composition is stable.

19. The composition of claim 16, wherein the anti-cancer agent is selected from the group consisting of anthracyclines, platinum complexes, mitoxantrone, nemorubicin, mitomycin C, bleomycin, actinomycin D, irinotecan, 5-fluorouracil, sorafenib, sunitinib, regorafenib, brivanib, orantinib, linsitinib, erlotinib, cabozantinib, foretinib, tivantinib, fotemustine, tauromustine (TCNU), carmustine, cytosine C, cyclophosphonamide, cytosine arabinoside, paclitaxel, docetaxel, methotrexate, everolimus, PEG-arginine deiminase, a tegafur/gimeracil/oteracil combination, muparfostat, peretinoin, gemcitabine, bevacizumab and ramucirumab, floxuridine, GM-CSF, molgramostim, sargramostim, OK-432, interleukin-2, interleukin-4 and TNFalpha, .sup.125I-labeled anti-CEA (carcinoembryonic antigen) antibodies, microspheres loaded with one of the foregoing, radioelements and complexes of said radioelements with macrocyclic chelates, magnetic particles based on an iron compound and/or on a gadolinium chelate, radioactive microspheres, deoxyribonucleic acid sequences, ribonucleic acid sequences, and combinations thereof.

20. The composition of claim 16, wherein the anthracyclines are chosen from doxorubicin, epirubicin, nemorubicin, and idarubicin.

21. The composition of claim 16, wherein the complexes of nonionic macrocyclic chelate with a paramagnetic metal are selected from the group consisting of: Gd-HP-DO3A; Gd-BT-DO3A; a complex of formula (XI): ##STR00014## wherein: R.sub.1, R.sub.2, and R.sub.3 each represent COOH; X.sub.1, X.sub.2 and X.sub.3 independently represent L-Y, wherein L represents a C.sub.1-C.sub.3 alkylene group, and Y represents CONH.sub.2, CONR.sub.7R.sub.8, or NR.sub.7COR.sub.8 in which R.sub.7 represents H, a C.sub.1-C.sub.6 alkyl group, or a C.sub.1-C.sub.8 hydroxyalkyl group, and R.sub.8 represents a C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.8 hydroxyalkyl group, on the condition that at least one of R.sub.7 or R.sub.8 represents a C.sub.1-C.sub.8 hydroxyalkyl group; D represents CH or N; E represents CH or N; F.sub.1 represents CH or N; K.sub.1 to K.sub.12 independently represent H, (CH.sub.2).sub.jCH.sub.3 or (CH.sub.2).sub.iOH in which j=0 to 3 and i=1 to 3, or K.sub.3 or K.sub.4 with K.sub.5 or K.sub.6 and/or K.sub.7 or K.sub.8 with K.sub.9 or K.sub.10 form, with the carbon atoms to which they are bonded, a ring having 3 to 6 carbon atoms; and M represents an ion of a paramagnetic metal; an enantiomer of the complex of the formula (XI); a diastereoisomer of the complex of the formula (XI); and a mixture thereof.

22. The composition of claim 16, wherein the lipid phase further comprises a non-iodized oil selected from the group consisting of linseed oil, soybean oil, palm oil, coconut oil, castor oil, corn oil, cottonseed oil, peanut oil, sesame oil, sunflower oil, safflower oil, almond oil, olive oil, poppy oil, and an oil comprising or consisting of a mixture of fatty acid triglycerides of formula: ##STR00015## wherein R is an aliphatic chain comprising from 3 to 35 carbon atoms, with the proviso that more than 95% of said fatty acids are C8 and/or C10.

23. The composition as claimed in claim 16, wherein the surfactant has an HLB of 1 to 8.

24. The composition of claim 16, wherein the surfactant is polyglyceryl polyricinoleate or PEG-30 dipolyhydroxystearate.

25. The composition of claim 16, wherein the iodized oil comprises ethyl esters of iodized fatty acids of poppy oil or olive oil.

26. The composition of claim 16, wherein the size of the aqueous phase droplets is from 1 to 200 m.

27. The composition of claim 16 having a viscosity at 20 C. included from 100 to 200 mPa.Math.s and/or a viscosity at 37 C. included from 40 to 80 mPa.Math.s.

28. The composition of claim 21, wherein: L represents a (CH.sub.2).sub.n group with n=1 to 3; and/or R.sub.7 and R.sub.8 independently represent a C.sub.2-C.sub.6 hydroxyalkyl group; and/or K.sub.1 to K.sub.12 represent H.

29. The composition as claimed in claim 21, wherein R.sub.7 and R.sub.8 independently represent CH.sub.2CH.sub.2OH, CHOHCH.sub.2OH, CH(CH.sub.2OH).sub.2, (CH.sub.2).sub.m(CHOH).sub.pCH.sub.2OH in which m=1 to 3 and p=1 to 4, or C(CH.sub.2OH).sub.3.

30. A method for the treatment of cancer or metastases thereof, the method comprising administering the composition of claim 1.

31. A method for preparing a composition as claimed in claim 1, the method comprising: a) mixing the surfactant in the iodized oil; and b) mixing the solution obtained in step a) with the aqueous solution comprising the anti-cancer agent and the densifying agent.

Description

EXAMPLE 1

1. Preparation of Compositions in the Form of an Emulsion According to the Invention

[0177] 1.1. Emulsions of Lipiodol and of Anthracycline

[0178] 50 mg of doxorubicin (Actavis) were reconstituted in 2.5 ml of gadobutrol (also called Gd-BT-DO3A and sold under the commercial name Gadovist 1 mmol/ml by the company Bayer). After manual stirring for 30 seconds for good dissolution, the solution obtained was removed with a 20 ml luer lock syringe. This syringe was then placed on a three-way tap.

[0179] PGPR (1% w/v total, 100 mgInterchim) was dissolved in 7.5 ml of Lipiodol by manual stirring.

[0180] The oil obtained was removed with a 20 ml luer lock syringe, which was also placed on the three-way tap at 90 C. 34 passes, i.e. 17 back-and-forward motions, at medium force were carried out, beginning with the water into the oil.

[0181] For these emulsions, the volumes of the aqueous phase and of the lipid phase chosen were respectively 2.5 ml (i.e. 25% v/v) and 7.5 ml (i.e. 75% v/v). The aqueous phase/lipid phase ratio was .

[0182] Other emulsions were prepared: [0183] by introducing no densifying agent, or [0184] by replacing the Gadovist densifying agent with the complex of nonionic macrocyclic chelate with a gadolinium ion of formula below

##STR00011##

(called compound X below) or by replacing it with a densifying agent chosen from the iodinated contrast products (Xenetix 250, Xenetix 300 (300 mg of iodine/ml), Iopamiron 350, Iopamiron 300, Iomeron 300, Ultravist 300 or Omnipaque 240), [0185] by replacing the Gadovist densifying agent with a mixture composed of 30% by volume of ProHance and of 70% by volume of Xenetix 350, [0186] by replacing the surfactant PGPR with Cithrol DPHS (PEG-30 dipolyhydroxystearate).

[0187] For the Cithrol DPHS, dissolution was obtained by using ultrasound (Vial tweeter, 345 s) or by heating the iodized oil.

[0188] Verification of the Sense of the Emulsion:

[0189] Once the emulsion had been prepared, the sense thereof was verified by means of a simple visual test. Two bottles were prepared: one with physiological saline and the other with iodized oil (Lipiodol).

[0190] A drop of freshly prepared emulsion was added to each of the two bottles. The drop dispersed in the bottle of Lipiodol and did not disperse in the bottle of physiological saline; the emulsion was therefore indeed a W/O (water-in-oil) emulsion.

[0191] The red doxorubicin droplets were clearly visible in a yellow background of oil. The size of the aqueous phase droplets was evaluated using an optical microscope.

[0192] The principal emulsions prepared are described in the following table:

TABLE-US-00005 Sizes of the Proportion of Nature of the Nature of the aqueous Visual Product Nature of the surfactant used anti-cancer densifying phase stability number surfactant used (% w/v) agent used agent used droplets observed* E1 PGPR 1% Doxorubicin None 5-40 m Phase separation <5% at 24 h E2 PGPR 1% Doxorubicin Iobitridol** 5-20 m No phase separation at 24 h E3 PGPR 1% Doxorubicin Gadobutrol 5-10 m No phase (Gadovist) separation at 24 h E4 PGPR 1% Doxorubicin Compound X 5-10 m No phase (see formula separation above) at 24 h E5 PGPR 1% Doxorubicin 30% by 5-10 m No phase volume of separation gadoteridol at 24 h (ProHance) and 70% by volume of Iobitridol***** E6 PGPR 1% Doxorubicin Iobitridol*** 5-20 m No phase separation at 24 h E7 PGPR 1% Doxorubicin Iopamidol**** 5-10 m No phase E8 PGPR 0.5%.sup. Doxorubicin Iobitridol** 5-20 m separation E9 PGPR 0.3%.sup. Doxorubicin Iobitridol** 5-20 m at 24 h E10 PGPR 1% Mitomycin C Iobitridol** 5-10 m No phase E11 PGPR 1% Epirubicin Iobitridol** 5-20 m separation E12 PGPR 0.7%.sup. Idarubicin Iobitridol** 2-10 m at 24 h E13 Cithrol DPHS 1% Doxorubicin None 5-20 m Phase separation <5% at 24 h E14 Cithrol DPHS 1% Doxorubicin Iobitridol** 5-20 m No phase E15 Cithrol DPHS 1% Doxorubicin Iobitridol*** 5-20 m separation at 24 h *at ambient temperature (20 C.) **Xenetix 250 ***Xenetix 300 ****Iopamiron 250 *****Xenetix 350

[0193] These various emulsions prepared using a surfactant of formula (I), various anti-cancer agents and densifying agents of different nature, all demonstrated a stability in accordance with expectations.

[0194] The replacement of the iodinated contrast products such as iobitridol and iopamidol, used as densifying agents, with complexes of nonionic macrocyclic chelate with a paramagnetic metal ion does not influence the stability of the emulsions obtained. By virtue of this replacement, it is possible to monitor the efficacy of the emulsions in accordance with the invention thus obtained by means of magnetic resonance imaging techniques such as MRI.

2. Comparison with Emulsions not in Accordance with the Invention

[0195] Emulsions according to the same protocol as that specified in paragraph 1.1 or a slightly different protocol (the differences compared with the protocol of paragraph 1.1 are indicated in the table below: the respective volumes of the aqueous and lipid phases are calculated on the basis of their ratio) were prepared using either a concentration of PGPR not in accordance with the invention, or surfactants having a low or high HLB, such as surfactants of the Span family (fatty acid esters of sorbitan), or surfactants of the HCO family (hydrogenated and ethoxylated castor oil), such as HCO-10 (HLB=6.5) or HCO-60 (HLB=14) supplied by the company Nikko Chemicals, surfactants having a high HLB of the Cremophor family (glycerol polyethylene glycol ricinoleate), of the Tween family (polyoxyethylene fatty acid esters of sorbitan) or of the Pluronics family (block copolymers based on ethylene oxides and propylene oxide, sold by BASF), and the Cithrol PG32IS surfactant having a low HLB (HLB=6.7).

[0196] Densifying agents other than the complexes of nonionic macrocyclic chelate with a paramagnetic metal ion were tested, such as a complex of ionic chelate with a gadolinium ion (Gd-DTPA, sold under the name Magnevist) or PVP (polyvinylpyrrolidone), glycerol, or else dextran T40 (Sigma), but the maximum amounts that can be used do not make it possible to approach the density of an iodized oil such as Lipiodol.

[0197] The principal emulsions prepared are described in the following tables:

[0198] Emulsions Prepared with a Surfactant in Accordance with the Invention but Using a Surfactant Concentration which does not Comply and/or a Densifying Agent which does not Comply

TABLE-US-00006 Aqueous Product Nature of the Proportion of Nature of the anti- phase/lipid number surfactant used surfactant used cancer agent used phase ratio Droplet sizes Observations E16 PGPR 0.2% by weight Doxorubicin 1/3 Heterogeneity: W/O emulsion relative to the 50 mg in 2.5 ml of small drops Phase total volume of Xenetix 250 and coarser separation the emulsion drops of 20- 50 m E17 PGPR 0.2% by weight Doxorubicin 1/3 Heterogeneity: W/O emulsion relative to the 50 mg in 2.5 ml of small drops Phase total volume of physiological saline and coarser separation the emulsion drops of 20- 50 m E18 PGPR 5% by weight Epirubicin 1/1 Heterogeneity: O/W emulsion relative to the 50 mg in 5 ml of drops ranging total phase total volume of water supplemented from 5 to 50 m separation in the emulsion with 290 mg of 2 hours glucose (5.8% w/v) E19 PGPR 1% by weight Doxorubicin 1/3 Heterogeneity W/O emulsion relative to the 50 mg in 2.5 ml Phase total volume of of Magnevist separation the emulsion

[0199] Emulsions Prepared with a Surfactant and/or a Densifying Agent not in Accordance with the Invention and/or in Aqueous Phase/Lipid Phase Ratios which do not Comply:

TABLE-US-00007 Proportion of surfactant used by weight relative to the Aqueous Product Nature of the total volume of Nature of the anti- phase/lipid number surfactant used the emulsion cancer agent used phase ratio Droplet sizes Observations E20 Span 80 1% (also tested: 2 ml of a solution 1/4 or 1/3 Aggregates of W/O emulsion 0.5% and 0.8%) of doxorubicin droplets of 50- Phase 50 mg in 10 ml of 100 m separation Xenetix 250 whatever the aqueous phase/lipid phase ratio and the proportion of surfactant used E21* Span 80 1% 2 ml of a solution 1/4 Heterogeneity: W/O emulsion of doxorubicin small drops Phase 50 mg in 10 ml of and coarser separation physiological drops of 20- saline supplemented 50 m with dextan T40 at 2.5 g/50 ml E22* Cremophor EL 0.5% 2 ml of a solution 1/4 2-5 m W/O emulsion of doxorubicin Phase 50 mg in 10 ml of separation physiological saline supplemented with dextan T40 at 3 g/50 ml E23 Tween 80 0.1% Doxorubicin 1/1 10 m O/W emulsion (50 mg in 5 ml of Slight phase Xenetix 250) separation at 24 h E24* Tween 80 0.1% or 0.01% Doxorubicin 1/1 10-100 m O/W emulsion (50 mg in 5 ml of physiological saline supplemented with Tween 80 and 1% of PVP) E25* Tween 80 0.01% Doxorubicin 1/1 100-300 m O/W emulsion HCl in 5 ml of Immediate glycerol at 2.5% phase separation E26 Mixture of 0.5% Doxorubicin 1/3 Heterogeneity Non- Tween 80 (50 mg in 2.5 ml of emulsified and Xenetix 250) unstable Span 80 0.5% system E27 CITHROL 1% Doxorubicin 1/3 Not W/O emulsion PG32IS (50 mg in 2.5 ml of measurable Instant and Xenetix 250) violent phase separation E28 Pluronic 5% Doxorubicin 1/4 10-100 m O/W emulsion L101 (50 mg in 2.5 ml of Partial phase water) separation before 18 hours E29 HCO-10 1% Doxorubicin 1/3 W/O emulsion (50 mg in 2.5 ml of Phase Xenetix 250) separation in 10 hours E30 HCO-60 1% Doxorubicin 1/3 W/O emulsion (50 mg in 2.5 ml of Rapid phase Xenetix 250) separation E31 HCO-60 1% Doxorubicin 1/1 O/W emulsion (50 mg in 5 ml of physiological saline) *emulsion prepared with a densifying agent not in accordance with the invention: PVP (polyvinylpyrrolidone), Dextran T40, ioxaglic acid (Hexabrix) or glycerol

[0200] Emulsions Prepared without Surfactant and/or without Densifying Agent:

TABLE-US-00008 Nature of the Aqueous Product anti-cancer phase/lipid number agent used phase ratio Droplet sizes Observations E32* Doxorubicin 1/1 10 m O/W emulsion (50 mg in 5 ml of physiological saline) E33 Doxorubicin 1/1 10 m O/W emulsion (50 mg in 5 ml of Xenetix 250) E34* Doxorubicin 1/3 or 1/2 Not W/O emulsion (50 mg in 2.5 ml of measurable Immediate Xenetix 250) (greater than phase 200 m) separation E35* Doxorubicin 1/4 10 m O/W emulsion (50 mg in 3 ml of Very viscous physiological and thick saline) emulsion: not usable in a clinical context E36 Epirubicin 1/1 10 m O/W emulsion (50 mg in 5 ml of Iopamiron 250) E37* Epirubicin 1/1 10-20 m O/W emulsion (50 mg in 5 ml of physiological saline) *emulsion prepared without densifying agent

[0201] Span 80 (Croda) is sorbitan monooleate. Tween 80 (Croda), also called polysorbate 80, is PEG-20 sorbitan monooleate. Cremophor EL (BASF) has, as chemical name: polyoxyl 35 Castor Oil. Cithrol PG32IS is polyglyceryl-3-diisostearate. It is not therefore branched like the surfactants of formula (I). HCO-10 (HLB=6.5) bears the INCl name: PEG-10 HYDROGENATED CASTOR OIL, while HCO-60 (HLB=14) bears the INCl name: PEG-60 HYDROGENATED CASTOR OIL.

[0202] Spans other than Span 80 (HLB=4.3) were tested: Span 20 (HLB=8.6), Span 65 (HLB=2.1), Span 83 (HLB=3.7) and Span 85 (HLB=1.8). The emulsions prepared with these surfactants all underwent phase separation immediately after they were prepared. The tests carried out with Pluronic compounds (BASF) were also not conclusive since it was not possible to prepare an emulsion with these compounds.

[0203] Thus, all of the comparative emulsions obtained exhibited either insufficient stabilities, or a sense not in accordance with the invention, or were not visible (or were insufficiently visible) by magnetic resonance imaging techniques such as MRI.