Carrier as multifunction system

Abstract

The present invention relates to a liposome comprising a core and at least one lipid bilayer # imgabs0 # comprising glycerolipid of general formula (I) wherein the core comprises a hydrophilic anti-cancer drug, antibacterial drug or other hydrophilic drug and / or the lipid bilayer comprises a hydrophobic anti-cancer drug, antibacterial drug or other hydrophobic drug. The invention also relates to a gel of glycerolipid of general formula (I) wherein the glycerolipid molecules form lipid nanotubes. Furthermore, the present invention relates to pharmaceutical compositions comprising said liposome or gel and at least one pharmaceutically acceptable excipient, which compositions can be used as antimicrobial agents (e.g., antibacterial agents or antiviral agents) or for treating cancer and / or preventing or reducing cancer metastasis, the invention also relates to a method for preparing the lipidosome containing the glyceride of the general formula (I) and a method for preparing the gel.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and particularly to a novel transporter (carrier) which combines multiple pharmacological activities and can be used for treating and / or preventing proliferative diseases. Background Art

[0002] The role of SK3 channels in cancer cell migration in vitro has been scientifically validated, as described in the published literature of multiple international journals (Potier et al., Mol Cancer Ther 2006, 5, 2946 - 53; Chantome et al., Exp Cell Res 2009, 315, 3620 - 30; Potier et al., Biochem Biophys Res Commun 2010, 397, 42 - 7; Girault et al., Current Cancer Drug Targets, 2011, 11, 1111 - 25; Sevrain et al., Med.Chem.Commun., 2012, 3, 1471 - 1478; Sevrain et al., Organic & Biomolecular Chemistry, 2013 Jul 21; 11(27):4479 - 87; Chantome et al., Cancer Research, 2013 Aug 1; 73(15):4852 - 61; Potier - Cartereau et al., Rev Physiol Biochem Pharmacol. 2022; 183:157 - 176). SK3 channels are potassium channels belonging to the SKCa family. Inhibiting SK3 reduces the migration ability of cancer cells, while the restoration experiment of SK3 can restore the migration ability of cancer cells, and forced expression of SK3 will increase the migration ability of cancer cells that do not express this channel. This channel is expressed in certain types of cells (such as brain cells, smooth muscle), and participates in the regulation of cell excitability by controlling cell secretion and muscle contraction. When SK3 is expressed in cancer cells, it doubles the cell's ability to migrate and invade through a matrix similar to the physiological extracellular matrix. Therefore, cancer cells seem to hijack the physiological function of SK3 in excitable cells to promote their migration ability. This channel forms a complex with the Orai1 calcium channel in cholesterol - rich nanodomains (Chantome et al., Cancer Research, 2013 Aug 1; 73(15):4852 - 61) and is associated with SigmaR1 (Gueguinou et al., Oncogene. 2017 Jun 22; 36(25):3640 - 3647).This complex in cholesterol-rich nanodomains has not been observed in non-tumor cells or excitable cells and is specific to cancer cells (Gueguinou et al., Biochim Biophys Acta. 2014 Mar 6; pii: S0167-4889(14)00083-4; Potier-Cartereau et al., Rev Physiol Biochem Pharmacol. 2022; 183:157-176). This novel non-physiological function of SK3 led us to speculate that the SK3 channel might be involved in the formation of metastatic tumors. Thus, using a mouse model of metastatic breast cancer (orthotopic xenograft), it was found that the SK3 channel does not regulate the development of the primary tumor but promotes the development of metastasis, mainly bone metastasis - there is a direct link between calcium-activated SK3 and the high calcium concentration found in the bone microenvironment (Chantome et al, Cancer Research, 2013 Aug 1; 73(15):4852-61). Recently, we found that the expression of SK3 is controlled by the epithelial-mesenchymal transition transcription factor Zeb1, which leads to enhanced calcium influx and increased cancer cell migration (Figiel et al., Cancers (Basel). 2019 Nov 18; 11(11):1814). In addition, in vitro and in organoid cultures of human biopsies, hypoxia (a recognized cancer hallmark associated with disease aggressiveness) significantly upregulated the expression of Zeb1 and the SK3 channel (Bery et al., Int J Mol Sci. 2020 Jul 6; 21(13):4786). Additionally, data on SK3 polymorphisms in breast cancer patients suggest that this might increase patients' sensitivity to the neurotoxicity of taxane drugs, and using an SK3 blocker during taxane treatment might be an interesting way to prevent this neurotoxicity (Rua et al., Clin Cancer Res; 2018, 24(21); 5313-20).

[0003] Proof-of-concept of this new mechanism that promotes migration in vitro and in vivo and cancer progression was also demonstrated by the identification of a new molecule, Ohmline (1-O-hexadecyl-2-O-methyl-sn-glycero-3-lactose), which inhibits SK3 channel activity and which is also the subject of the present invention (Girault et al., Curr Cancer Drug Targets. 2011 Nov;11(9):1111-25; Jaffres et al., Pharmacol Ther. 2016 Sep;165:114-31).

[0004] Ohmline belongs to a new class of alkyl ether glycolipids developed to inhibit cancer-related metastatic processes. This compound targets the cell membrane and specifically affects K+ channels located in nanodomains, leading to a decrease in membrane potential and thus reducing the migratory capacity of cancer cells (Chantome et al, Cancer Research, 2013 Aug 1; 73(15):4852-61). It has been found that Ohmline can specifically reduce the migratory capacity of SK3-expressing cancer cells with 50% efficiency at a concentration of 10 nM. It should be noted that at concentrations up to 1 μM, the reduction in the migration of some cancer cells is not attributed to the inhibition of SK3. The inhibitory effect of Ohmline on cancer cell migration has been observed in all tested SK3-expressing cells, including breast cancer MDA-MB-435s and prostate cancer PC3 cells (Chantome et al, Cancer Research, 2013 Aug 1; 73(15):4852-61; Figiel etal., Cancers (Basel). 2019 Nov 18; 11(11):1814). In addition, it has been found that Ohmline can inhibit TGFβ-induced upregulation of Zeb1 and calcium influx in prostate cancer cells under normoxic and hypoxic conditions (Figiel et al., Cancers (Basel). 2019 Nov 18; 11(11):1814; (Bery et al., Int J Mol Sci. 2020 Jul 6; 21(13):4786), and can also inhibit paclitaxel-induced calcium influx, which is more significant in cells with short CAG repeats expressing the SK3 gene than in cells with long repeats (Rua et al., Clin Cancer Res 2018; 24(21); 5313-20). Ohmline can dissociate the SK3 channel complex in cholesterol-rich nanodomains by reducing Akt phosphorylation and regulating the action of anti-EGFR monoclonal antibodies in colorectal cancer cells (Chantome et al, Cancer Research, 2013 Aug 1; 73(15):4852-61; Gueguinou et al., Oncotarget. 2016 Jun 14; 7(24):36168-36184).

[0005] Although the SK3 channel has been found to specifically participate in the development of bone metastasis, its inhibitor Ohmline can not only reduce the development of bone metastasis, but also reduce lung and lymph node metastasis. When SK3-expressing cells are injected intravenously, Ohmline can delay the development of bone metastasis (Chantome et al, Cancer Research, 2013 Aug 1; 73(15):4852-61). Interestingly, it has been found that this lipid can displace cholesterol, moving its hydroxyl group away from the main binding site and forcing it to rearrange with other lipid groups (Herrera et al., ACS Omega. 2017 Oct 31; 2(10):6361-6370).

[0006] Therefore, Jaffres et al. (Pharmacol Ther. 2016 Sep; 165:114-31) proposed Ohmline and its analogs as novel anti-metastatic drugs. This study revealed that synthetic alkyl lipids (such as Ohmline, 1-O-hexadecyl-2-O-methyl-rac-glycero-3-β-lactose) can target cell membranes and reduce cell migration / invasion, thereby inhibiting the development of metastasis. In addition, international application WO 2011 / 101408 "Methods for preventing or treating cancer metastasis" discloses glycerolipid compounds, their synthesis methods, and their uses for preventing cancer metastasis, in which OHM (Ohmline) is referred to as compound JPH1701. The invention disclosed in WO 2011 / 101408 aims to provide compounds with anti-metastatic activity, which can be regarded as true metastasis inhibitors, that is, those with anti-metastatic activity but having no effect on the primary tumor. Metastasis inhibitors are alleged to act through a mechanism different from that of tumor inhibitors. In fact, there are few anti-cancer drugs with proven therapeutic activity against the primary tumor that also have anti-metastatic activity. The invention disclosed in WO 2011 / 101408 provides certain glycerolipid compounds (which are also ether lipids, such as Ohmline) for preventing cancer metastasis or for treating against the occurrence of metastasis. This activity is attributed to the inhibition of the SK3 / KCa2.3 channel, which has been experimentally confirmed, demonstrating the anti-metastatic activity of glycerolipids (including Ohmline), while showing their specificity for the metastatic process without affecting primary tumor growth. The experiment was carried out by incubating glycerolipid compounds with cell cultures or administering them to animal models, using DMSO and ethanol as carriers. The pharmaceutical compositions disclosed in WO2011 / 101408 contain 0.01% to 99.99% by weight of the glycerolipids of the present invention and one or more excipients. Liposome encapsulation is one of the delivery systems proposed in WO 2011 / 101408 (although not used in the experiment) for glycerolipids such as Ohmline.

[0007] Another important property of the glycerolipids disclosed in WO 2011 / 101408 is their low cytotoxicity, an effect similar to the anti-metastatic activity and requiring the presence of the glycerol backbone. The low cytotoxicity is an important difference from a similar compound, edelfosine, which is an alkyl glycerophospholipid known to have anti-angiogenic and possible anti-invasive effects (possibly mediated by the phosphocholine group), but also has high toxicity.

[0008] In fact, other phospholipids with ether group structures and anti-cancer activity also show toxicity problems. This is the case for some lipids described in European patent application EP0785773A1. The invention disclosed in this application relates to liposomes, which are defined as structures self-assembled from one or more lipid bilayers, each bilayer surrounding an aqueous chamber and containing two opposing monolayers of amphiphilic lipid molecules that comprise a polar (hydrophilic) head group region covalently linked to one or two non-polar (hydrophobic) acyl chains, so that in an aqueous medium, the lipid molecules are induced to rearrange such that the polar head groups face the aqueous medium while the acyl chains are reoriented to the interior of the bilayer.

[0009] This definition is similar to the definitions found in general publications on liposomes and other lipid particles, such as the review by Sharma et al. on liposomes as drug delivery systems (Sharma et al, The Pharma Tutor Magazine, 2018; 6(2): 50 - 62), which states that liposomes consist of a bilayer or multilamellar lipid body, containing or having phospholipids (the main component of liposomes, defined as amphiphilic molecules with a hydrophilic head and a hydrophobic tail, usually composed of two acyl chains of 10 - 24 carbon atoms) and cholesterol, surrounding an aqueous chamber, with a diameter of 0.01 - 5.0 μm, formed when phospholipids are hydrated in an excess of aqueous medium or aqueous solution. More specifically, the liposome preparation method involves four basic stages: a) drying the lipids from an organic solvent, b) dispersing the lipids in an aqueous medium; c) purifying the resulting liposomes; d) analyzing the final product. It is also mentioned that size determines the metabolism of liposomes in the body, with larger liposomes (diameter greater than 0.1 μm) being cleared from the circulation faster than smaller liposomes and being more stable in the circulation when they have a hydrophilic coating. Other publications (e.g., "The Signifi cant Difference Between Micelles and Liposomes", https: / / biologywise.com / difference-between-micelles-liposomes) describe the comparison between liposomes and micelles (lipid nanoparticles with only one layer of amphiphilic / amphipathic molecules), stating that liposomes are larger and have a higher drug - loading capacity, emphasizing that their formation increases above a specific temperature (referred to as the transition temperature in the case of liposomes), and commenting that micelles are mainly formed by surfactant molecules such as detergents, emulsifiers, various wetting agents, and certain copolymers, while liposomes are formed by phospholipid molecules and cholesterol.In other general publications, such as "Lipids Aggregates in Water-Micelles and Liposomes" (https: / / bio.libretexts.org / Bookshelves / Biochemistry / Fundamentals_of_Biochemistry_(Jakubowski_and_Flatt) / 01%3A_Unit_I-Structure_and_Catalysis / 10%3A_Lipids / 10.02%3A_Lipids_Aggregates_in_Water_-Micelles_and_Liposomes), the importance of the number of non-polar chains in the structures formed by amphiphilic molecules in water is emphasized. It is pointed out that some single-chain amphiphilic molecules, when added to water, form a monolayer on the water surface and form micelles when exceeding their solubility, while double-chain amphiphilic molecules, on the contrary, form bilayers instead of micelles. Liposomes are spherical bilayers that enclose an aqueous compartment.

[0010] The liposomes disclosed in European Patent Application EP0785773A1 particularly comprise a lipid bilayer composed of at least two lipids: a lipid with a derivatized head group and a glycerolipid containing an ether group, the backbone of which is similar to the glycerolipid of WO 2011 / 101408, but different from the said glycerolipid in that it is also a phospholipid because the group R3 attached to the carbon atom at the opposite end of the ether group must be a group with the formula R3 = R5-P(O)2-O-R6 and is thus a phospholipid. The lipid with a derivatized head group is preferably phosphatidylethanolamine-dicarboxylic acid. In a preferred embodiment, the liposome lipid bilayer contains ether lipids and lipids with derivatized head groups, as well as sterols (preferably cholesterol) and neutral lipids (preferably phosphatidylcholine). Additional bioactive agents may be present in the liposomes. The inclusion of lipids with derivatized head groups is because it can buffer the toxicity of ether lipids when present in the liposome bilayer and is said to generally be an amphiphilic lipid, including one or more hydrophobic acyl chains and a polar head group connected by a chemical group. Such a composition produces single-layer or multi-layer liposomes, less than 200 nm, preferably greater than 50 nm. These liposomes are used for the treatment of cancer and inflammatory diseases.

[0011] Ohmline has passed the chemical optimization stage because the optimization of its synthesis has been completed (Alkyl etherlipids, ion channels and lipid raft reorganization in cancer therapy. Jaffrès PA, Gajate C, Bouchet AM, Couthon-Gourvès H, A, Potier-Cartereau M, Besson P, Bougnoux P, Mollinedo F, Vandier C. Pharmacol Ther. 2016 Sep;165:114-31. doi: 10.1016 / j.pharmthera.2016.06.003. Epub 2016 Jun 8). Ohmline showed good tolerance and marked safety in rodents (e.g., no tissue damage was observed by pathologists). Preclinical studies conducted in vitro did not show adverse biological activity.

[0012] The potential toxicity and genotoxicity of Ohmline were examined by using the Ames test and the micronucleus test. The Ames test showed no mutagenic potential, and the micronucleus test showed a weak positive effect only at 100 μM. These findings suggest that Ohmline is non-toxic and non-genotoxic at in vitro concentrations below 10 μM. Lower in vitro toxicity was also observed in cancer cell lines MDA-MB-231, HCT116, PC3, HaCaT, and fibroblasts: 70% to 100% of the cells remained viable after treatment with 25 μM Ohmline for 48 hours, compared with almost 0% viability after treatment with Edelfosine (a structural analogue of Ohmline but with phosphorylcholine instead of lactose at the sn3 position) under the same conditions (ImPACcell-Rennes) (Girault et al., Curr Cancer Drug Targets. 2011 Nov;11(9):1111-25; Jaffres et al., Pharmacol Ther. 2016 Sep;165:114-31; Potier et al., Br J Pharmacol. 2011 Jan;162(2):464-79).

[0013] Finally, preliminary pharmacokinetic studies found that Ohmline was absorbed by the test tissues (stomach, intestine, kidney, liver, heart, and brain) and remained in the brain two weeks after discontinuation of dosing. In addition, integration of Ohmline into primary tumors and bone metastases tissues was also observed (Chantome et al, Cancer Research, 2013 Aug 1;73(15):4852-61).

[0014] Currently, there is no specific treatment for the development of metastases, nor is there a treatment to prevent metastases. Only conventional treatments such as radiotherapy, chemotherapy, or hormone therapy (for some cancers) are used to treat metastases. Palliative treatments for pain or bone resorption (in cases of bone metastases) are also provided.

[0015] However, several clinical trials for the treatment of metastatic castration-resistant prostate cancer have emerged, but the extension of median survival is modest (measured in months). A phase III clinical trial showed that the use of Denosumab (an anti-RANKL drug developed by Amgen) could extend the bone metastasis-free survival by 4.2 months, but had no effect on overall survival (Smith et al. Lancet, 2012; 379:39-46). Denosumab does not target cancer cells, but rather targets osteocytes by preventing osteocyte lysis induced by metastatic development. The anti-RANKL antibody seems to be complementary to anti-SK3 treatment, as it does not target specific markers of tumor cells, which may be similar to conventional non-targeted chemotherapy, but the estimated treatment cost is much higher.

[0016] Therefore, there is still a need in the prior art for new treatment methods for metastatic development and treatment methods for preventing metastasis, which should target the SKCa channels, especially the SK3 channel, have a lower cost compared to conventional non-targeted chemotherapy, and minimize the impact on other targets and adverse reactions. Ideally, the product designed for this purpose can also be used to treat other diseases, such as microbial infections. Summary of the Invention

[0017] The main object of the present invention is to develop a novel transporter (also known as a carrier) with multiple pharmacological activities. It has a dual function: it can act as a carrier for compounds with pharmacological effects (usually called therapeutic drugs) while having therapeutic activity itself.

[0018] To achieve this object, the applicant has designed and constructed a transporter (Ohmline) that already has pharmacological activity itself. The structure of the transporter (or carrier) allows the insertion of hydrophobic and hydrophilic drugs and confers diverse therapeutic properties on the transporter (carrier). The multivalent activity of this transporter (carrier) is a major breakthrough in the prior art.

[0019] This achievement is attributed to the discovery that Ohmline can spontaneously form liposomes in polar solutions (such as aqueous solutions) without the presence of any known liposome-forming compounds in the solution, such as phospholipids or any other amphiphilic compounds with polar heads and double hydrophobic chain tails. This is a surprising discovery because, unlike the liposome-forming lipids in previous publications (such as European Patent Application EP0785773A1), Ohmline does not have the characteristic structure of a compound that forms liposomes in polar solutions. The lipids in EP0785773A1 are phospholipids and another lipid with a derivatized head group (preferably also a derivatized phospholipid), and are preferably used together with sterols (such as cholesterol) and neutral lipids (such as phosphoethanolamine). Based on its structure, Ohmline was originally expected to form lipid nanoparticles of the micelle type rather than liposomes. However, as Figure 1 shown, Ohmline can form liposomes in which sugar molecules are exposed on the surface and in the liposome core defined by the lipid bilayer.

[0020] This property is also surprising to those skilled in the art of International Patent Application WO2011 / 101408. The structure of Ohmline and its anti-metastatic activity were disclosed in this document, but when testing the activity of Ohmline in this application, the compound was used directly rather than in any assembled structure; there was also no mention or implication of the possibility of forming liposomes or any other nanoparticles in this application document.

[0021] However, surprisingly, providing a mixture containing an aqueous buffer, an appropriate concentration of Ohmline, and (if desired) another compound (preferably an anti-cancer drug) and stirring the mixture at a temperature above 40 °C (the transition temperature of Ohmline) results in the formation of liposomes containing Ohmline in the lipid bilayer. Thus, the present invention discloses, at least in one aspect, a process for preparing Ohmline liposomes. A specific example of such a process can be a method for preparing Ohmline liposomes carrying another anti-cancer drug, anti-microbial drug, or any other drug (which will be in the core if hydrophilic and in the lipid bilayer if hydrophobic). Additionally, if the Ohmline concentration is high (equal to or preferably higher than 3 mg / ml), a gel structure is formed in which Ohmline molecules exist in the form of nanotubes.

[0022] The property of Ohmline to self-assemble into liposomes above its transition temperature (40 °C - 44 °C, a temperature range specific to Ohmline) enables these nanoparticles to serve as carriers for other drugs or pharmacologically active compounds, whether hydrophilic compounds (which can be encapsulated in the liposome core) or hydrophobic compounds (incorporated in the lipid bilayer), and possess the advantages of liposomes, such as a greater loading capacity for other compounds compared to micelles, and the property of easily delivering the payload to cells by fusing with the cell membrane. Nevertheless, Ohmline nanoparticles are highly fusogenic with the cell membrane, which aids their entry into cells and the release of any payload in the cytoplasm. Additionally, as mentioned above, when the Ohmline concentration exceeds a certain threshold, Ohmline molecules can also form gels in which glycerolipid molecules form lipid nanotubes; as shown in the examples. In vitro, Ohmline nanotubes form a sustained-release depot, which can also serve as a delivery system for in situ cancer treatment. From the mentioned in vitro experiments, it can be understood that by dilution, Ohmline nanotubes turn into nanoparticles that will autonomously release Ohmline and any encapsulated drugs, as if self-releasing. Thus, Ohmline may exist in two different but reversible structural arrangements (nanotube gels or liposomes), which may be useful for its delivery.

[0023] The special structure of Ohmline liposomes, with a surface composed of sugars (specifically, monosaccharides and disaccharides composed of lactose groups in this example), confers additional advantages as it helps prevent the clearance of liposomes from the circulation and enables them to be directed to and bind to lectin receptors, which are specifically expressed in tumor and / or inflammatory environments.

[0024] Furthermore, Ohmline liposomes are formed from compounds that are themselves pharmacologically active, so Ohmline liposomes have a dual function: they are therapeutically active themselves and can simultaneously serve as carriers for other drugs. In this regard, Ohmline liposomes have an advantage over the liposomes of EP0785773A1 as their basic structure may be formed solely from the molecules of one compound (Ohmline). Another remarkable and advantageous property is the low cytotoxicity of Ohmline, while EP0785773A1 acknowledges the toxicity of the anticancer glycerolipid, one of the components of its liposomes, and makes buffering this toxicity the main objective of the invention disclosed in that European patent application.

[0025] Notably, experiments on Ohmline liposomes have shown that Ohmline administered in the form of nanoparticles can reduce the size of the primary tumor. This is another beneficial and surprising effect, especially considering the content of WO2011 / 101408, where experiments showed that Ohmline administered in monomeric form does not affect the primary tumor. Moreover, its anti-metastatic effect is not only maintained but even enhanced.

[0026] In addition, the Applicant has also found that Ohmline in the form of liposomes or gels can also be used to treat microbial infections, again taking advantage of the dual function of Ohmline: its ability as a drug carrier (preferably an antimicrobial drug in this case) and as an antimicrobial drug itself. This is confirmed in the examples of the present application, where it is shown that the Ohmline hydrogel depot exhibits antibacterial activity. This activity appears to be related to the K+ channels that Ohmline molecules specifically target, which are essential for bacterial intracellular homeostasis and communication, and are important not only for quorum sensing but also for biofilm formation. It is suggested that biofilm formation may account for more than 65% of all infections, and Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Escherichia coli are considered to be the most difficult bacteria to treat due to biofilm formation. In this regard, the characteristic of the ordered structure of Ohmline to expose sugars on its surface enables it to bind to lectin receptors and be used for anti-adhesion therapy against biofilm-forming bacteria, because lectins have a known role in establishing and maintaining biofilms, and the knockout or inhibition of biofilm-mediated lectins leads to the disruption of biofilm integrity (Inhülsen et al., Microbiologyopen 2012, 1(2), 225-242). Moreover, as described above, the regulation of the biophysical behavior of Ohmline makes this material an excellent tool for precision medicine and can be used to provide personalized treatment, especially in cases where two or more antimicrobial agents (or antimicrobial drugs in general) are required to act synergistically.

[0027] In this text, an antimicrobial agent refers to a compound or composition that can kill microorganisms, reduce or hinder their growth, reduce or hinder their entry into cells or their spread in a host, or reduce or hinder the formation of structures (such as biofilms) that facilitate organ colonization. Microorganisms can be bacteria, protozoa, microscopic fungi (such as yeasts) or even viruses, and the host can be any animal (including invertebrates and vertebrates, with humans being an example) or any plant.

[0028] Accordingly, a first object of the present invention relates to a liposome comprising a core and at least one lipid bilayer, the lipid bilayer comprising a glycerolipid of general formula (I):

[0029]

[0030] wherein the core comprises a hydrophilic anticancer drug, a hydrophilic antimicrobial agent or any other hydrophilic drug, and / or the lipid bilayer comprises a hydrophobic anticancer drug, a hydrophobic antimicrobial agent or any other hydrophilic drug. Preferably, the core comprises a hydrophilic anticancer drug and / or the lipid bilayer comprises a hydrophobic anticancer drug.

[0031] As described above, the lipid bilayer may be free of phospholipids or any other amphiphilic lipid having a polar head and a double-chain non-polar tail. However, it may contain hydrophobic compounds such as anticancer drugs, antimicrobial agents or any other hydrophobic drugs.

[0032] In another embodiment, the lipid bilayer consists of a glycerolipid of general formula (I). This may occur when the lipid bilayer does not contain any anticancer drugs, antimicrobial agents and other drugs other than Ohmline itself.

[0033] Thus, in one embodiment, the lipid bilayer consists only of glycerolipid molecules of general formula (I), and the core comprises a hydrophilic anticancer drug, a hydrophilic antimicrobial agent or any other hydrophilic drug.

[0034] A second object of the present invention relates to a glycerolipid gel of general formula (I). wherein the glycerolipid molecules form lipid nanotubes

[0035] A third object of the present invention relates to a pharmaceutical composition comprising the liposome or gel described above and at least one pharmaceutically acceptable excipient, which compositions can be used as antibacterial or antiviral agents, or for the treatment of cancer or infections, or for the prevention or reduction of cancer metastasis.

[0036] Another object of the present invention is to use the liposome of the first object, the gel of the second object or the pharmaceutical composition of the third object as a therapeutic agent.

[0037] In a preferred embodiment, it is used for the treatment of bacterial, viral or fungal infections. In a specific embodiment, the infection is a biofilm-associated bacterial infection. For example, the infection can be caused by Gram-negative or Gram-positive bacterial species, such as Escherichia coli or Staphylococcus aureus, or Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus epidermidis or Pseudomonas aeruginosa. In another embodiment compatible with the first three embodiments, the liposome or gel contains an antimicrobial drug.

[0038] In a preferred embodiment, the liposome, gel or pharmaceutical composition is used for the treatment or prevention of cancer. In a specific embodiment of this embodiment, it is used for the inhibition or reduction of primary tumor growth. In another specific embodiment, it is used for the prevention or reduction of cancer metastasis. In another embodiment compatible with all the embodiments mentioned in this paragraph, the liposome or gel contains an anticancer drug as a supplement to the glycerolipid of general formula (I) (Ohmline itself).

[0039] Other objects of the present invention relate to methods for preparing liposomes and methods for preparing gels containing glycerolipids of general formula (I), wherein the formation of one or the other structure depends on the concentration of Ohmline. The method for preparing liposomes includes at least the following steps: (i) providing a mixture that contains an aqueous buffer, at least one anticancer drug, antimicrobial drug or any other drug, and a glycerolipid of general formula (I), wherein the concentration of the glycerolipid of general formula (I) in the mixture is equal to or lower than 3 mg / ml; preferably from 0.04 mg / ml to 3 mg / ml; (ii) stirring the mixture at a temperature higher than 40 °C; wherein, when the anticancer or antimicrobial drug or other drug is a hydrophobic compound, the anticancer or antimicrobial drug or other drug is added together with the mixture of the glycerolipid of general formula (I) obtained in the previous step; when the anticancer or antimicrobial drug or other drug is a hydrophilic compound, the anticancer or antimicrobial drug or other added drug is incorporated into the aqueous buffer. The mixture in step (i) can be free of any phospholipids or any other amphiphilic lipids having a polar head and a double-stranded non-polar tail.

[0040] Accordingly, the method for preparing the gel comprises at least the following steps: (i) providing a mixture comprising an aqueous buffer, at least one anti-cancer or anti-microbial drug or any other drug, and a glycerolipid of general formula (I), wherein the concentration of the glycerolipid of general formula (I) in the mixture is higher than 3 mg / ml; (ii) stirring the mixture at a temperature higher than 40 °C; wherein, when the anti-cancer or anti-microbial drug or the other added drug is a hydrophobic compound, the anti-cancer or anti-microbial drug or the other added drug is added together with the mixture of the glycerolipid of general formula (I) obtained in the previous step; when the anti-cancer or anti-microbial drug or the other added drug is a hydrophilic compound, the anti-cancer or anti-microbial drug or the other drug is incorporated into the aqueous buffer. Description of the Drawings

[0041] Figure 1 : Schematic diagram of the bilayer Ohmline structure, showing sugar molecules exposed on the surface and in the core, while acyl chains are located inside the bilayer Ohmline itself. This schematic diagram shows how the molecular assembly of the bilayer can be achieved by single acyl chain molecules such as Ohmline. Two independent Ohmline molecules above the bilayer can be seen in the figure, and each molecule shows the expected arrangement according to its position in the layer. An enlarged version of the same Ohmline molecular formula is also shown in the figure, where the sugar ring is simplified to a simple hexagonal ring, and all members of each ring are projected on the same plane.

[0042] Figure 2 : Fluorescence microscopy image (A) and phase contrast (B) of Ohmline MLV (1 mg / ml) labeled with the hydrophobic probe BODIPY 493 / 503. White scale bar = 5 μm. The visible size range of Ohmline multilamellar vesicles is between 500 nm and 2.5 μm.

[0043] Figure 3: Transmission electron microscopy images of Ohmline LUVs prepared by the extrusion method ( Figure 3A ) or the sonication method ( Figure 3B ), with concentrations of 100 μM and 20 μM respectively.

[0044] Figure 4: Phase contrast and fluorescence microscopy images of CF encapsulated in Ohmline MLV. (λex = 492 nm, λem = 517 nm). Among them, Figure 4A are the phase contrast, fluorescence and merged fields of view respectively, white scale bar = 2.5 μm. Figure 4B are the phase contrast and fluorescence respectively, white scale bar = 10 μm.

[0045] Figure 5: Optical microscope image of Ohmline MLV labeled with di4ANNEPDHQ. Excitation wavelength (λexcit) = 488 nm; emission wavelength (λemiss) = 520 to 700 nm.

[0046] Figure 6 : Time series: (A) = time 0, (B) = in progress, (C) = end. In all pictures, the left test tube contains a methanol solution probe as a control that is completely miscible with water. The Ohmline gel phase remains stable and immiscible in water, forming a reservoir.

[0047] Figure 7 : (A) corresponds to Figure 6 48 hours after injection of the Ohmline gel described. In all pictures, the left test tube contains a methanol solution probe control that is miscible with water. (B) corresponds to 72 hours after injection, at which time the reservoir has been converted to liposomes containing the probe.

[0048] Figure 8 : Electron micrograph of Ohmline (10 mg / ml) after heating (above 40 °C) and vortex cycling homogenization. A few milliliters of the sample were deposited on a grid for electron microscopy measurement and stained with uranyl acetate. Black scale bar in (A) = 100 nm, black scale bar in (B) = 200 nm.

[0049] Figure 9 : Stimulated emission depletion (STED) microscope image of Ohmline hydrogel (4 mg / ml) co-encapsulated with tetramethylcarbodicyanine and glycerolipid structures. (λex = 550 nm, λem = 567 nm). White scale bar in the image = 2 μm.

[0050] Figure 10: (A) Bacterial growth curves showing Escherichia coli (left panel) and Staphylococcus aureus (right panel), showing the control group (data represented by circles) and bacteria treated with Ohmline (data represented by squares). (B) Fluorescence microscopy images of Escherichia coli (left two panels) and Staphylococcus aureus (right two panels), showing propidium iodide internalization due to Ohmline treatment (red in the original image, indicated by arrows in the modified image). (C) Bar graph showing the percentage of bacterial growth inhibition caused by Ohmline treatment (Escherichia coli: left bar; Staphylococcus aureus: right bar), calculated relative to the values of the control group. (D): Shows the effect of Ohmline treatment on the bacterial replication time indicated below the X-axis (in each pair of bars, the Ohmline-treated group is the left bar; the control group is the right bar). Detailed implementation mode

[0051] The present invention is based on the principle of inhibiting the activity of plasma membrane proteins. Therefore, this does not require therapeutic molecules to enter the interior of cancer cells, thereby minimizing the impact on other targets and adverse effects.

[0052] Ohmline and its derivatives are considered anti-metastatic molecules targeting the SK3 channel in the prior art.

[0053] Ohmline and its derivatives are lipid molecules retained on the plasma membrane and have a simple chemical formula.

[0054] Ohmline is a glycosyl-glycerol-ether lipid with the chemical formula (1-O-hexadecyl-2-O-methyl-sn-glycero-3-lactose), and thus contains sugar in its structure: two linked monosaccharides (glucose and galactose) form the disaccharide lactose. Its synthesis can be easily accomplished using standard organic chemistry protocols. Ohmline showed no acute toxicity when administered to small animals at a dose of 15 mg / Kg for up to 4 months. Ohmline is the first specific inhibitor of the K+ channel SK3: it is more potent for the SK3 channel than for the SK1 channel; it does not affect the IKCa / SK4 channel and has only a slight but not significant effect on the SK2 channel. This is the first SKCa channel blocker that can be used to distinguish between SK2 and SK1 / SK3 channels. The IC50 of this compound is 300 nM, and it does not displace the bee venom extract peptide (apamin, an SKCa channel blocker that cannot distinguish between SK1, SK2, and SK3 channels) from the SKCa channel, and has no effect on the targets of non-specific edelfosine (an alkyl lipid with phosphocholine instead of lactose at the glycerol sn3 position), such as the receptors of protein kinase C (PKC), platelet-activating factor (PAF), and lysophosphatidic acid (LPA), and non-cancer cells. (Girault et al., Curr Cancer Drug Targets 2011 Nov;11(9):1111-25)

[0055] The mechanism of action of Ohmline is novel compared to other ion channel blockers: it does not act as a pore blocker like apamin and most ion channel modulators, but rather blocks the activation of the SK3 channel after it is incorporated into the plasma membrane, thereby affecting the intracellular membrane. Thus, this lipid can be used to reduce the activity of various SK3 channels: SK3 channels sensitive and insensitive to apamin (an embryonic SK3 channel insensitive to apamin has been discovered), as well as potentially non-ion channel-forming SK3 proteins (this situation has been found in certain potassium ion proteins eag1 in cancer cells, but further studies on SK3 proteins are still needed) (Downie et al., J Biol Chem, 2008;283(52):36234-40).

[0056] In addition, the Ohmline molecule, as a monomer, specifically acts on the K+ channels that are crucial for bacterial cell homeostasis and communication (quorum sensing / biofilm). Regardless of the metabolic state of the bacteria, bacterial K+ channels play a key role in membrane potential regulation, intercellular and interspecies communication, and memory formation (Prindle et al., Nature 2015, 527(7576), 59-63). Membrane potential is crucial not only for sub-second functions such as motility and cell respiration but also for processes that require minutes to hours, such as cell division and intercellular communication. Therefore, targeting K+ channels is key to disrupting bacterial survival, interaction, adaptation, and communication under various environmental conditions (Samuel et al., Trends in Microbiology 2021; 29(10), 894-907). Previous studies have shown that biofilm-associated infections account for more than 65% of all infections (Wang et al., Biofilms and Microbiomes 2023, 9(1), 63), and biofilms are defined as structured aggregates attached to living or inert surfaces, composed of mutually adherent microbial cells and surrounded by a self-produced extracellular polymeric matrix (Wu et al., International Journal of Oral Science, 2014; 7:1-7). Interestingly, potassium channel deletion strains show defective biofilm development, highlighting the relevance of this membrane protein as a target for antimicrobial systems (Diggle et al., PloS one 2013; 8(5):e60993).

[0057] The applicant has found that when Ohmline is dispersed in an aqueous medium, it can form a nano-liposome structure. This latest discovery has fundamentally changed the positioning of Ohmline, which should be regarded not only as a drug but also as a drug transporter. To the applicant's knowledge, this duality has not been proven until now.

[0058] In recent years, the application of lipid nanoparticle technology in cancer treatment has grown exponentially. Due to their small size, lipid nanoparticles decorated with specific antibodies or any protein capable of recognizing specific targets can accumulate in tumors, with limited effects on other healthy parts of the body, and have the ability to improve treatment efficacy and reduce the major adverse reactions of chemotherapy. Currently, 19 commercial versions of lipid nanoparticles have been approved for treatment, and another 17 are in the clinical stage (Allen and Cullis, Adv. Drug Rev. 2013; 65:36-48; Liu et al., Int. J. Nanomedicine, 2013; 8:3309-3319).

[0059] Thus, the liposomes and gels provided by the present invention combine the anti-metastatic and anti-cancer activities of Ohmline, as well as its ability to transport various drugs. This unique property is related to the unique ability of Ohmline to form a lamellar phase in different nanostructures.

[0060] In addition, the Ohmline liposomes and gels of the present invention have antimicrobial activity per se, in addition to being able to act as carriers for other antimicrobial drugs. Thus, Ohmline, as an organized structure, can be used for anti-adhesion therapy (AAT) and antimicrobial drug delivery. Although not wishing to be bound by any theory, AAT may be attributed to the exposure of sugar molecules on the surface of Ohmline nanoparticles or nanotube gels (for example, see Figure 1 ), and thus is able to bind to lectin receptors and prevent microbial adhesion. This is consistent with previous findings that lectins play a role in the establishment and maintenance of biofilms (Diggle et al., Environmental Microbiology 2006, 8(6), 1095-1104), and that the knockout or inhibition of biofilm-mediated lectins leads to the disruption of biofilm integrity (Inhüsen et al., Microbiologyopen 2012; 1(2):225-242; Tielker et al., Microbiology 2005; 151(5):1313-1323). In addition, the biophysical behavior regulation of Ohmline makes this material an excellent tool for precision medicine, which can be used to provide personalized treatment, especially in cases where two or more antimicrobial drugs need to act synergistically.

[0061] The above are all beneficial and surprising effects of the structures formed by Ohmline.

[0062] For most nanoparticles, it is the associated drug that confers its specific activity. For example, liposomes have been successfully used both in vitro and in vivo, but the lipid is only a transporter and does not have specific pharmacological activity.

[0063] Surprisingly, the applicant has found that: liposomes and gels containing Ohmline are excellent carriers for transporting various drugs, and exhibit favorable properties based on the antimicrobial, anti-metastatic and / or anti-cancer activities of Ohmline itself and its ability to transport various drugs. This unique property is related to the unique ability of Ohmline to adopt different lipid structures.

[0064] Therefore, the present invention provides:

[0065] A liposome comprising a core and at least one lipid bilayer containing a glycerolipid of general formula (I):

[0066]

[0067] Wherein, the core contains a hydrophilic anti-cancer drug, a hydrophilic anti-microbial drug or any other hydrophilic drug, and / or the lipid bilayer contains a hydrophobic anti-cancer drug, a hydrophobic anti-microbial drug or any other hydrophobic drug.

[0068] In a preferred embodiment, the core contains a hydrophilic anti-cancer drug and / or the lipid bilayer contains a hydrophobic anti-cancer drug.

[0069] In another embodiment compatible with the foregoing, the lipid bilayer does not contain phospholipids or any other amphiphilic lipid having a polar head and a double-chain non-polar tail. In this embodiment, the lipid bilayer may contain a hydrophobic anti-cancer drug, an anti-microbial drug or any other hydrophobic drug.

[0070] In another embodiment, the lipid bilayer is composed of glycerolipid molecules of general formula (I). This is compatible with the following embodiments because the core may contain a hydrophilic anti-cancer drug, a hydrophilic anti-microbial drug or any other hydrophilic drug.

[0071] In other embodiments of the present invention, the hydrophilic anti-cancer drug, anti-microbial drug or other hydrophilic drug is encapsulated within the core.

[0072] Preferably, by microscopic measurement, the diameter of the liposome is from 0.01 μm to 20 μm, preferably from 0.05 to 5 μm, more preferably from 2.5 to 5 μm.

[0073] In another preferred embodiment of the present invention, the hydrophilic or hydrophobic drug contained in the liposome is an anti-cancer drug, which is selected from the following list: aromatase inhibitors, anti-estrogen, anti-androgen, gonadorelin agonists, topoisomerase 1 inhibitors, topoisomerase 2 inhibitors, microtubule active agents, alkylating agents, anthracyclines, corticosteroids, IMiDs, protease inhibitors, IGF-1 inhibitors, CD40 antibodies, Smac mimetics, FGF3 regulators, mTOR inhibitors, HDAC inhibitors, IKK inhibitors, P38MAPK inhibitors, HSP90 inhibitors, akt inhibitors, anti-tumor drugs, anti-metabolic drugs, platinum-containing compounds, lipid or protein kinase targeting drugs, protein or lipid phosphatase targeting drugs, anti-angiogenic drugs, cell differentiation inducing drugs, bradykinin 1 receptor antagonists, angiotensin II antagonists, cyclooxygenase inhibitors, heparinase inhibitors, lymphokine inhibitors, cytokine inhibitors, bisphosphonates, rapamycin derivatives, anti-apoptosis pathway inhibitors, apoptosis pathway agonists, PPAR agonists, Ras isozyme inhibitors, telomerase inhibitors, protease inhibitors, metalloproteinase inhibitors and aminopeptidase inhibitors, cell statics, cytotoxic drugs, taxanes, anti-tumor antibiotics, topoisomerase II inhibitors, topoisomerase I inhibitors, microtubule interacting agents, antibodies, anti-angiogenic agents, COX-2 inhibitors, hormonal drugs, thymidylate synthase inhibitors, anti-metabolic drugs, alkylating agents, farnesyl protein transferase inhibitors, signal transduction inhibitors, EGFR kinase inhibitors, EGFR antibodies, C-abl kinase inhibitors, hormonal therapy combination drugs, aromatase inhibitors and combinations thereof.

[0074] In a more preferred embodiment of the present invention, the anti-cancer drug is a taxane drug selected from the list consisting of docetaxel, paclitaxel, tesetaxel, and cabazitaxel; preferably, the taxane drug is docetaxel.

[0075] The present invention also provides a gel of glycerolipid of general formula (I):

[0076]

[0077] Among them, the glycerolipid molecules form lipid nanotubes.

[0078] Preferably, the gel is a hydrogel (see Figure 8 and 9 ).

[0079] According to one embodiment of the present invention, the liposome or gel can be used as a carrier for DNA, siRNA, RNA, and antibodies.

[0080] According to one embodiment of the present invention, the liposome or gel can be used as a carrier for antibodies or nanobodies.

[0081] Preferably, the liposome or gel can be used as a drug, for example, for the treatment and / or prevention of diseases.

[0082] Therefore, another aspect of the present invention relates to the use of the liposome or gel (preferably a hydrogel) of the foregoing aspect as a drug.

[0083] The drug can be used as an antimicrobial drug (for example, an antibacterial drug, an antiviral drug). In particular, when the liposome or gel contains an antimicrobial drug (except for the glycerolipid Ohmline of general formula (I)), it can be used for the treatment of bacterial, viral, protozoal, or fungal infections, especially biofilm-related bacterial infections, whether caused by Gram-positive or Gram-negative bacteria.

[0084] The liposome or gel, or a pharmaceutical composition containing any of them, can also be used for the treatment and / or prevention of cancer or reducing cancer metastasis. For example, it can be used for primary tumor growth inhibition, or as described above, for preventing or reducing metastasis. When the liposome or gel or a pharmaceutical composition containing them is used for cancer treatment (whether for primary tumor growth inhibition or preventing or reducing metastasis), preferably the liposome or gel contains an anticancer drug other than Ohmline itself. The cancer can be breast cancer, head and neck cancer, gastric cancer, prostate cancer, non-small cell lung cancer, leukemia, skin cancer, melanoma, liver cancer, ovarian cancer, pancreatic cancer, lung cancer, kidney cancer, colon cancer, brain cancer (selected from glioma and glioblastoma).

[0085] The present invention also provides a pharmaceutical composition for therapeutic use, which comprises a liposome or gel and at least one pharmaceutically acceptable excipient and / or an adjuvant. The pharmaceutically acceptable excipient and / or adjuvant is selected from the following list: diluents, buffers, preservatives, stabilizers, surfactants, solvents, disintegrants, antioxidants, binders, lubricants, and colorants, etc.

[0086] The pharmaceutical composition of the present invention can be in oral, topical or injectable dosage forms.

[0087] The excipients can be selected from any excipients known to those skilled in the art and are selected according to the route of administration to be used and the intended therapeutic objective.

[0088] Oral preparations can be, for example, tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs.

[0089] The compositions for oral use can be prepared by any method known in the pharmaceutical art, and these compositions can contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents, and preservatives to provide a pharmaceutically elegant and palatable preparation.

[0090] The tablets contain a mixture of the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients can be, for example: inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binding agents such as starch, gelatin, or acacia; and lubricating agents such as magnesium stearate, stearic acid, or talc.

[0091] The tablets can be uncoated or can be coated using known techniques to delay their disintegration and absorption in the gastrointestinal tract, thereby providing a longer-lasting effect. For example, sustained-release materials such as glyceryl monostearate or glyceryl distearate can be used.

[0092] The oral preparation can also be made into hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin; or into soft gelatin capsules, in which the active ingredient is mixed with a water or oily medium, such as peanut oil, liquid paraffin or olive oil.

[0093] The aqueous suspension contains the active ingredient mixed with excipients suitable for the preparation of aqueous suspensions. These excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; the dispersing agent or wetting agent can be natural phospholipids, such as lecithin, or the condensation product of an alkyl oxide and a fatty acid, such as polyoxyethylene stearate, or the condensation product of ethylene oxide and a long-chain fatty alcohol, such as heptadecaethyleneoxycetanol, or the condensation product of ethylene oxide and a partial ester from a fatty acid and a hexitol, such as polyoxyethylene sorbitol monooleate, or the condensation product of ethylene oxide and a partial ester from a fatty acid and a hexitol anhydride, such as polyethylene sorbitan monooleate.

[0094] The aqueous suspension can also contain one or more preservatives, such as ethyl, or n-propyl, p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0095] The oily suspension can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil (arachis oil), olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. The oily suspension may contain a thickening agent such as beeswax, hard paraffin or acetyl alcohol. Sweetening agents and flavoring agents as described above may be added to provide a palatable oral preparation.

[0096] These compositions may be preserved by adding an antioxidant such as ascorbic acid. Dispersible powders and granules for preparing an aqueous suspension by the addition of water, comprise the active ingredient mixed with a dispersing or wetting agent, a suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are as described above. Other excipients may also be present, such as sweetening agents, flavoring agents and coloring agents. Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol or sucrose. These preparations may also contain a demulcent, preservatives, and flavoring and coloring agents.

[0097] For injectable forms, the carrier generally consists of water and auxiliary substances necessary to render it compatible with physiological conditions, such as pH-regulating agents, tonicity agents and the like.

[0098] For external forms, the excipient selection depends on the dosage form expected in each case, preferably liquid or semi-solid. Liquid preparations include aqueous solutions, hydroalcoholic solutions, lotions and the like. Semi-solid preparations may be ointments, creams or gels. Any of these preparations may require the use of preservatives, antioxidants, chelating agents, emulsifying agents, pH regulating systems, osmotic regulating substances, demulcents, bioadhesive polymers, gel forming substances and the like.

[0099] Among them, the preservative can be selected from the following substances, for example: sodium benzoate, ascorbic acid, parabens, bronopol, methylisothiazolinone and their mixtures. The antioxidant can be selected, for example: butylated hydroxytoluene (BHT), hydroxyanisole and tocopherol and their derivatives. The chelating agent can be selected from the following groups, for example: ethylenediaminetetraacetic acid (EDTA), ethyleneglycol bis-(2-aminoethyl)-tetraacetic acid (EGTA) and citric acid or its salts. The pH adjustment system can be selected, for example: phosphate buffer solution and citrate buffer solution. The gel-forming substance can be selected, for example: Carbopols, Poloxamers, Carboxymethylcellulose (CMC), Hydroxyethylcellulose (HEC), Hydroxypropyl-methyl (HPMC), Methylcellulose (MC) and their mixtures.

[0100] In one embodiment of the present invention, the stabilizer or surfactant is an anionic / cationic lipid, and / or its weight concentration in the composition is 5-45% w / w.

[0101] Preferably, the pharmaceutical composition according to the present invention can be prepared as a vaccine carrier, for example, as an RNA or DNA vaccine. Preferably, the stabilizer or surfactant of the RNA or DNA vaccine is a cationic lipid.

[0102] The pharmaceutical composition according to the present invention is suitable for the treatment or prevention of cancer, particularly colorectal cancer, lung cancer, pancreatic cancer and gastric cancer, or for treating a combination selected from colorectal cancer, lung cancer, gastric cancer, breast cancer, head and neck cancer, prostate cancer, non-small cell lung cancer, leukemia, skin cancer, liver cancer, ovarian cancer, pancreatic cancer, lung cancer and kidney cancer.

[0103] The pharmaceutical composition containing the liposome or gel of the present invention as a drug can be used to prevent or reduce the occurrence of metastasis in any body tissue or any body organ. The uses of the glycerolipids of general formula (I) include preventing or treating the occurrence of metastasis in various tissues and organs, including the ovary, uterus, kidney, liver, lung, bone tissue (e.g., leg bones including the femur, arm bones, spine including the thoracic and lumbar vertebrae, pelvis), spleen, lymph nodes, colon, breast, brain, prostate, pancreas, and skin.

[0104] The present invention can be used in combination with other anti-cancer treatment methods, such as chemotherapy, cryotherapy, hyperthermia, radiotherapy, etc.

[0105] The present invention also includes a method for preparing liposomes, which at least includes the following steps:

[0106] i) Provide a mixture, which includes

[0107] an aqueous buffer;

[0108] at least one anti-cancer or antibacterial agent or any other drug; and

[0109] the glycerolipids of general formula (I);

[0110]

[0111] wherein, the concentration of the glycerolipids of general formula (I) in the mixture is equal to or lower than 3 mg / ml; preferably from 0.04 mg / ml to 3 mg / ml;

[0112] ii) Stir the mixture at a temperature higher than 40 °C, preferably from 40 °C to 80 °C.

[0113] wherein, when the anti-cancer or antibacterial agent or other added drug is a hydrophobic compound, the anti-cancer or antibacterial agent or other added drug is added to the mixture in step i) together with the glycerolipids of general formula (I), and

[0114] wherein, when the anti-cancer or antibacterial agent or other added drug is a hydrophilic compound, the anti-cancer agent is included in the aqueous buffer.

[0115] In a preferred embodiment of the present invention, the hydrophobic compound is dissolved in a polar solvent and dried together with Ohmline. These two compounds: the hydrophobic compound (anti-cancer or antibacterial agent or other drug) to be co-encapsulated with Ohmline and Ohmline are dissolved in a polar buffer, dried with a hydrogen stream to form a film, and then hydrated with an aqueous buffer.

[0116] In a preferred embodiment of the present invention, the glycerolipid of general formula (I) is in powder form, first dissolved in a solvent or solvent mixture, such as chloroform / methanol (preferably 3:1), and then dried under vacuum and / or in a nitrogen atmosphere to form a film, which is rehydrated in a buffer such as phosphate buffered saline (pH = 7.6).

[0117] The liposome preparation method further includes step (iii) of extruding the mixture obtained in step (ii), preferably by membrane or sonication treatment.

[0118] The present invention also includes a method for preparing a gel, which at least includes the following steps:

[0119] i) Providing a mixture, which includes:

[0120] An aqueous buffer;

[0121] At least one anti-cancer or antibacterial agent or any other drug; and

[0122] The glycerolipid of general formula (I);

[0123]

[0124] Wherein, the concentration of the glycerolipid of general formula (I) in the mixture is higher than 3 mg / ml, and more preferably, the concentration of the glycerolipid of general formula (I) in the mixture is 3 to 25 mg / l;

[0125] ii) Stirring the mixture at a temperature higher than 40 °C, preferably between 40 and 80 °C

[0126] Wherein, when the anti-cancer or antibacterial agent or other added drug is a hydrophobic compound, the anti-cancer or antibacterial agent or other drug is added to the mixture together with the glycerolipid of general formula (I), and

[0127] Wherein, when the anti-cancer or antibacterial agent or other added compound is a hydrophilic compound, the anti-cancer agent is included in the aqueous buffer.

[0128] Preferably, the aqueous buffer is selected from the list of phosphate buffered saline (PBS), sodium chloride buffer, sterile water and their combinations.

[0129] According to the present invention, data indicate that Ohmline forms liposomes with the ability to act as a drug delivery vehicle. Interestingly, these Ohmline liposomes themselves (empty carriers) act as a novel anti-tumor agent, possibly due to the enhanced permeability and retention (EPR) effect, which results in far more accumulation in tumor tissues than in normal tissues.

[0130] This new property of Ohmline clearly shows that this lipid can not only prevent metastasis, but also act as an anti-tumor agent itself. More importantly, it can carry and deliver another anti-tumor agent, showing a synergistic effect.

[0131] Therefore, these multiple properties have not been proven so far in the field of drug carriers. The ability to prevent cancer and metastasis, as well as deliver drugs, is a major innovative breakthrough in the technical field of the present invention, improving the effectiveness of chemotherapy.

[0132] Specific examples

[0133] The preparation of a novel transporter that combines multiple pharmacological activities is described below. To achieve this goal, the applicant designed and prepared a transporter (Ohmline) that has pharmacological activity by itself. The structure of this carrier allows the insertion of hydrophobic and hydrophilic drugs and confers diverse therapeutic properties on the same carrier.

[0134] To demonstrate the superior properties of the transport carrier of the present invention, the applicant performed biophysical characterization of the Ohmline lipid phase, encapsulated hydrophilic probes (carboxifluorescein) and hydrophobic drugs such as taxane drugs (docetaxel), and conducted in vitro and in vivo tests. In addition, the present invention also shows the in vivo test results of a preparation named TAXOM prepared according to the present invention for different types of cancers (such as breast cancer) and different docetaxel concentrations.

[0135] 1 - General liposome preparation method

[0136] The synthesis method of Ohmline is as previously described (Girault A, et al. New alkyl-lipid blockers of SK3 channels reduce cancer cell migration and occurrence of metastasis. Current cancer drug targets. 2011; 11: 1111-1125).

[0137] Dissolve Ohmline powder in a chloroform:methanol mixture (3:1), evaporate under a nitrogen stream, and dry overnight under vacuum to remove any residual solvent. Vortex the dried film in the buffer selected for each experiment at temperatures above 40 °C.

[0138] Hydrated Ohmline sheets detached during stirring and formed multilamellar vesicles (MLV); further extrusion or sonication can be used to prepare large unilamellar vesicles (LUV), and sonication may also yield small unilamellar vesicles (SUV).

[0139] 1.1 - Observation of MLV by optical microscope

[0140] Prepare Ohmline multilamellar vesicles labeled with the hydrophobic probe BODIPY (product D3922 from Thermo Fisher Scientific, https: / / www.thermofisher.com / order / catalog / product / D3922). The size range of the MLV is between 500 nm and 2.5 μm. Figure 2 Fluorescence microscopy images of Ohmline MLV at a concentration of 1 mg / ml labeled with BODIPY 493 / 503 are shown, with liposome sizes ranging from 500 nm to 2.5 μm.

[0141] 1.2 - Observation of LUV by electron microscope

[0142] Prepare LUV samples using different concentrations of Ohmline MLV by sonication or extrusion. Place a Formvar / carbon-coated nickel grid on the sample droplet for 5 minutes and rinse twice with water droplets. Then perform three consecutive counterstains using 2% uranyl acetate (Agar Scientific, Stansted, UK), and analyze under a transmission electron microscope (JEOL 1011, Tokyo, Japan). Prepare unilamellar vesicles with a size of 100 nm by extrusion through a polycarbonate membrane with a 100 nm pore size using a microextruder from Avanti Polar Lipids. Prepare unilamellar vesicles of approximately 50 nm by treatment in an ultrasonic water bath at temperatures above 50 °C.

[0143] Figure 3A and Figure 3B Show Ohmline samples with final dilution concentrations of 100 μM and 20 μM on the grid, respectively.

[0144] 2 - Hydrophilic probe carboxyfluorescein encapsulated in Ohmline multilamellar liposomes

[0145] Mix the Ohmline dry film with a concentration of 0.8 mg / ml (1.23 mM) with 400 μM of carboxyfluorescein (CF) (6-carboxyfluorescein, Sigma Aldrich product C0662, https: / / www.sigmaaldrich.com / ES / es / product / sigma / c0662) in PBS buffer (pH = 7.6). Encapuslate the probe within the Ohmline structure by multiple freeze-thaw cycles and vortexing. Use a PD-10 pre-packed column (Cytiva, https: / / www.cytivalifesciences.com / en / us / shop / chromatography / prepacked-columns / desalting-and-buffer-exchange / sephadex-g-25-in-pd-10-desalting-columns-p-05778) and Sephadex G-25M to separate free and encapsulated CF, and the encapsulation of CF (green) can be observed.

[0146] Figure 4A and Figure 4B Show the phase contrast microscope and fluorescence microscope images (excitation wavelength λex = 492 nm, emission wavelength λem = 517 nm) of Ohmline MLV encapsulated with carboxyfluorescein.

[0147] The Ohmline MLV loaded with CF shows that the conventional size of the multilayer particles is between 2.5 and 5 μm.

[0148] 3 - Lipophilic probe

[0149] Any lipophilic (or hydrophobic) molecule will insert into the lipid bilayer of the Ohmline liposome, which can be illustrated by the fluorescent probe di-4-ANNEPDHQ (Thermo Fisher Scientific product D36802, https: / / www.thermofisher.com / order / catalog / product / D36802). Incubate the Ohmline liposome with 2 μl of the DMSO stock solution of di-4-ANNEPDHQ at a lipid / probe ratio of 100. Final concentration: Ohmline is 1.23 mM and di-4-ANNEPDHQ is 12.3 μM.

[0150] Figure 5 Shows a microscopic image of Ohmline MLV labeled with di4ANNEPDHQ. Excitation wavelength (λexcit) = 488 nm, emission wavelength (λemiss) = 520 to 700 nm. In this case, the particle size of Ohmline liposomes into which a lipophilic probe has been inserted is between 1 and 5 μm.

[0151] 4 - Docetaxel encapsulated in Ohmline MLV

[0152] Docetaxel was purchased from Sigma-Aldrich (Sigma-Aldrich product 01885-5MG-F, https: / / www.sigmaaldrich.com / ES / es / search / 01885-5mg-f?focus=products&page=1&perpage=30&sort=relevance&term=01885-5mg-f&type=product). It was used to prepare liposomes and, as described below, was also used to prepare a formulation similar to the commercial drug TAXOTERE, which contains 20 mg of docetaxel / 0.5 ml of TWEEN 80 as an excipient, as well as 95% (v / v) ethanol and water (13 / 87 p / p).

[0153] Ohmline liposomes loaded with docetaxel are hereby named TAXOM. TAXOM concentration is usually expressed as a ratio (docetaxel nM / Ohmline μM), using the abbreviation TAX to represent TAXOM, DOX to represent docetaxel, showing that Ohmline liposomes fuse with cells and release their payload.

[0154] MLV Ohmline liposomes were prepared in PBS buffer (pH = 7.6) according to the method described in Section 1 above.

[0155] Ohmline was dissolved to form liposomes by hydrating the thin film, and docetaxel was inserted into the lipid bilayer at the ratio specified in each experiment. TAXOM particles are prepared and used on the same day.

[0156] 4.1 - In vitro cell activity test

[0157] Ohmline nanoparticles were used to deliver docetaxel to different cancer cell lines. Cell viability was determined using the tetrazolium reduction method (MTT, CyQUANTTM MTT Cell Viability Assay, Thermo Fisher Scientific product V13154, https: / / www.thermofisher.com / order / catalog / product / V13154?ef_id=4431670897231bd9f4af0436727d3335:G:s&s_kwcid=AL!3652!10!77240846955634!77240903742099&cid=bid_pca_iva_r01_co_cp1359_pjt0000_bid00000_0se_bng_nt_pur_con&msclkid=4431670897231bd9f4af0436727d3335# / V13154), which reflects the number of living cells. On day 0, 6500 cells were seeded in a 48-well plate. The conventional medium of cancer cells was added to each well. On day 1, the medium was aspirated and replaced with fresh medium of the formulation to be tested. The cells were cultured for up to 3 days.

[0158] At the end of the culture, the medium was aspirated and replaced with an MTT solution at a concentration of 0.5 mg / ml. Incubate at 37 °C for 30 - 45 minutes. After incubation, the supernatant was aspirated, and the formed crystals were dissolved with DMSO, gently shaken at room temperature, protected from light. Transfer 200 μl from each well to a 96-well plate and measure the absorbance at a wavelength of 570 nm (see Figure 8 A, 8B, and 8C).

[0159] Formulations:

[0160] Ohmline: Prepared in the MLV formulation at concentrations of 1 μM and 5 μM.

[0161] DOX: Dissolved in DMSO at concentrations of 0.5 and 1 nM.

[0162] TAX: In a chloroform:methanol mixture (3:1), Ohmline and docetaxel were co-dissolved at different molar ratios to prepare MLV:

[0163] TAX 0.5 / 1 = 0.0005 docetaxel / Ohmline

[0164] TAX 0.5 / 5 = 0.0001 docetaxel / Ohmline

[0165] TAX 1 / 1 = 0.001 docetaxel / Ohmline

[0166] TAX 1 / 5 = 0.0002 docetaxel / Ohmline

[0167]

[0168] Table 1. Percentage of viable tumor cells in different cancer cell lines

[0169] Under certain conditions, we observed a decrease in cell viability after treatment with Ohmline empty liposomes alone. The TAX formulation was always more effective than DOX, and higher efficiency was observed when the Ohmline content in TAX was higher (e.g., TAX = 0.5 / 5 was better than TAX = 0.5 / 1).

[0170] Empty Ohmline liposomes can be used as anti-tumor agents.

[0171] Ohmline liposomes loaded with docetaxel (TAX) are more effective than treatment with Ohmline or docetaxel alone (synergistic effect).

[0172] 4.2 - In vivo animal model

[0173] The researchers used 48 female NMRI nude mice (from Janvier Labs, France, https: / / janvier-labs.com / ), 4 weeks old, 12 mice per treatment. Cancer model: MDA-MB-435s breast cancer cells were inoculated into the mammary fat pad. These mice were housed and managed in the Inserm U892 laboratory at Nantes University, and the animal experiment license number was N°44565. Two weeks after inoculating the cancer cells, the mice were injected intravenously. Duration of the experiment: 2 months, for the detection of lung metastases.

[0174] Treatment protocol

[0175]

[0176] Formulation:

[0177] Group A / Control group: Given 150 mM NaCl by intravenous injection (i.v.) - 2 injections per week for 2 weeks

[0178] Group B / Ohmline group: Ohmline 15 mg / Kg = 3 mg / ml Ohmline in 150 mM NaCl, for 30 g mice, intravenous injection - 2 injections per week for 2 weeks

[0179] Group C / Docetaxel group: Taxotere formulation prepared in the applicant's laboratory: 5 mg / Kg, intravenous injection - twice a week for 2 weeks

[0180] Group D / TAXOM group: 15 mg / Kg (3 mg / ml Ohmline in 154 mM NaCl, same as group A) + 5 mg / Kg docetaxel (100 mg / ml DMSO stock solution - 1.5 μl), intravenous injection - twice a week for 2 weeks

[0181] The MLV of Ohmline and docetaxel were co - solubilized in a chloroform:methanol mixture to prepare MLV containing both components. Molar ratio: 0.4 docetaxel / Ohmline.

[0182] Treatment method Standardized mean reduction size Control group 0 Docetaxel - Taxotere 38 Ohmline MLV 39.5 TAX 44.5

[0183] Table 2. In vitro analysis results. Average reduction in tumor size under each formulation after normalization

[0184]

[0185] Table 3. In vitro analysis. Percentage of metastasis development for each formulation.

[0186] Conclusion:

[0187] The main contribution relative to the prior art is that the use of Ohmline liposomes alone has an equivalent effect to other drug formulations in reducing tumor size.

[0188] The anti - metastatic activity reaches the best effect in the TAX formulation, which means that the combination of Ohmline and docetaxel (even if the ratio we selected is not optimal) can improve the quality of treatment.

[0189] A very important finding is that 2 mice died after treatment with Taxotere, while no mice died when using Ohmline MLV or its combination formulation with docetaxel.

[0190] 5. General hydrogel preparation

[0191] Dissolve the Ohmline powder in a chloroform:methanol mixture (ratio 3:1), evaporate under a nitrogen stream, and dry overnight under vacuum to remove any residual solvent. Vortex the dried film with the buffer selected for each experiment above 40 °C. When the Ohmline concentration is higher than 3 mg / ml, the hydrated Ohmline becomes viscous.

[0192] Macroscopic appearance: Viscous hydrogel

[0193] Dissolve 4 mg of Ohmline (6.10 mM) and tetramethylindocyanine (61 μM, Sigma - Aldrich, product 42364: https: / / www.sigmaaldrich.com / ES / es / product / sigma / 42364) in an organic solvent (a chloroform:methanol mixture, in a 3:1 ratio) such that the molar ratio of Ohmline to the probe is 100:1. Evaporate the solvent under a nitrogen stream and dry overnight. Hydrate the film in miliQ water and vortex - mix at temperatures above 40 °C. Observe the appearance of the Ohmline gel in vitro and its ability to encapsulate the lipophilic probe (tetramethylindocyanine).

[0194] In vitro formability and degradability study

[0195] Use two transparent test tubes containing 1 ml of high - purity deionized (MilliQ - water) to observe the behavior of the Ohmline hydrogel. For the control group, dissolve tetramethylindocyanine in methanol. Dissolve the probe at the same final concentration in each test tube. Figure 6 Photographs A, B, C in show the time series. In all figures, the left - hand test tube contains the probe in methanol solution, which diffuses rapidly in the medium. However, the Ohmline gel injected into water is able to maintain its phase for several days. The accumulation of the probe within the gel can be clearly observed ( Figure 6 C). After 2 days, the gel remains at the bottom ( Figure 7 A), and gradually decreases. On the 3rd day, the gel becomes completely clear (transforms into a liposome solution that retains the probe) ( Figure 7 B).

[0196] It can be concluded that this hydrogel forms a sustained - release reservoir in vitro and may serve as a promising delivery system for in situ treatment of cancer and even treatment of infections.

[0197] Microscopic appearance: Nanotubes forming a viscous network

[0198] Prepare an Ohmline hydrogel at a concentration of 10 mg / ml and observe the samples using an electron microscope according to the same protocol as previously described for LUVs (large unilamellar vesicles). Figure 8 Shows an extensive network of nanotubes intersecting at various positions.

[0199] Microscopic appearance: Nanotubes as carriers of lipophilic probes

[0200] The same Ohmline hydrogel containing tetramethylcarboline at 4 mg / ml, characterized previously, was used to observe the nanotubes and their ability to carry probes. A STED microscope was used to improve the resolution. Figure 9 The same extensive nanotube network as observed by electron microscopy was shown, but this time the lipophilic probes could be seen incorporated into the lipid structures. The Ohmline gel can encapsulate hydrophobic drugs for in-situ drug delivery, avoiding the side effects associated with systemic drug circulation.

[0201] 6 - Antibacterial activity

[0202] Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 29923 were used as reference strains (ATCC: American Type Culture Collection), representing Gram-positive and Gram-negative bacteria, respectively. Bacterial cultures were grown using Mueller Hinton Broth medium from Britania Lab S.A. (www.britanialab.com).

[0203] All experiments were conducted by placing 20 μl of a 5 mg / ml Ohmline gel reservoir in a 96-well plate, in which 200 μl of bacteria were cultured with an initial OD value (600 nm) of 0.15. OD 600 values were recorded at multiple time points within 24 hours. The bacteria were cultured with shaking at 37 °C. All experiments treated the bacteria with Ohmline hydrogel reservoirs (100% Ohmline component), allowing Ohmline to be released into the culture medium (final concentration 0.5 mg / mL).

[0204] As Figure 10 shown, the Ohmline hydrogel reservoir exhibited direct antibacterial activity against both E. coli and S. aureus, with a minimal inhibitory concentration (MIC) of 0.5 μg / ml against S. aureus.

[0205] Microscopic images ( Figure 10 B) showed that the plasma membrane levels of both bacterial species were damaged, highlighting the role of this nanomaterial as a synergist that can increase the activity of other antibacterial agents and / or reverse antibiotic resistance.

[0206] More importantly, some preclinical data from animal models have confirmed that intravenous injection and oral administration of Ohmline nanomaterials in rats showed no toxicity at doses up to 500 mg / Kg. ADME (absorption, distribution, metabolism and excretion) studies also showed extensive distribution and tropism for tissues and organs. Ohmline also showed distribution to the brain (able to cross the blood-brain barrier).

Claims

1. A liposome, comprising: Core and a lipid bilayer comprising at least one glyceride of general formula (I): wherein the core comprises a hydrophilic anti-cancer drug, a hydrophilic antibacterial drug or other hydrophilic drug, and / or the lipid bilayer comprises a hydrophobic anti-cancer drug, a hydrophobic antibacterial drug or other hydrophobic drug.

2. The liposome according to claim 1, wherein, The core comprises a hydrophilic anti-cancer drug, and / or the lipid bilayer comprises a hydrophobic anti-cancer drug.

3. The liposome according to claim 1 or 2, wherein The lipid bilayer does not contain phospholipids or other amphiphilic lipids having a polar head and a double-chain non-polar tail.

4. The liposome according to any one of claims 1-3, wherein, The lipid bilayer consists essentially of the glyceride of general formula (I):

5. The liposome according to any one of claims 1-4, wherein the lipid bilayer is composed only of the glyceride molecules of general formula (I): and the core comprises a hydrophilic anti-cancer drug, a hydrophilic antibacterial drug or other hydrophilic drug.

6. The liposome according to any one of claims 1-5, wherein, The hydrophilic anti-cancer drug is encapsulated within the core.

7. The liposome according to any one of the preceding claims, wherein, Measured by optical microscopy, the liposome has a diameter of 0.01 to 20 microns, preferably 0.05 to 5 microns, more preferably 2.5 to 5 microns.

8. The liposome according to any one of the preceding claims, wherein, The liposome comprises an anti-cancer drug; the anti-cancer drug is selected from the following list: Aromatase inhibitors, anti-estrogens, anti-androgens, gonadotropin-releasing hormone agonists, topoisomerase 1 inhibitors, topoisomerase 2 inhibitors, microtubule-active drugs, alkylating agents, anthracyclines, corticosteroids, immunomodulators, protease inhibitors, IGF-1 inhibitors, CD40 antibodies, Smac mimetics, FGF3 regulators, mTOR inhibitors, histone deacetylase inhibitors, IKK inhibitors, P38MAPK inhibitors, HSP90 inhibitors, akt inhibitors, anti-tumor drugs, anti-metabolic drugs, platinum-containing compounds, drugs targeting lipid or protein kinases, drugs targeting protein or lipid phosphatases, anti-angiogenic drugs, drugs inducing cell differentiation, bradykinin 1 receptor antagonists, angiotensin II antagonists, cyclooxygenase inhibitors, heparinase inhibitors, lymphokine inhibitors, cytokine inhibitors, bisphosphonates, rapamycin derivatives, anti-apoptosis pathway inhibitors, apoptosis pathway agonists, PPAR agonists, Ras subtype inhibitors, telomerase inhibitors, protease inhibitors, matrix metalloproteinase inhibitors and aminopeptidase inhibitors, cell inhibitors, cytotoxic drugs, taxanes, anti-tumor antibiotics, topoisomerase II inhibitors, topoisomerase I inhibitors, microtubule-interacting drugs, antibodies, anti-angiogenic drugs, COX-2 inhibitors, hormonal drugs, thymidylate synthase inhibitors, anti-metabolic drugs, alkylating agents, farnesyl protein transferase inhibitors, signal transduction inhibitors, EGFR kinase inhibitors, anti-EGFR antibodies, C-abl kinase inhibitors, hormonal therapy combination drugs, aromatase inhibitors and combinations thereof.

9. The liposome according to the preceding claim, wherein, The anti-cancer drug is a taxane drug selected from the following list: docetaxel, paclitaxel, tesetaxel, cabazitaxel; preferably, the taxane drug is docetaxel.

10. A gel of a glyceride of the following general formula (I): Among them, The molecules of the glyceride self-assemble to form lipid nanotubes.

11. The gel according to the preceding claim, wherein, The gel is a hydrogel.

12. Use of the liposome or gel according to the preceding claims as a DNA or RNA carrier.

13. Use of the liposome according to any one of claims 1-9 or the gel according to any one of claims 10-11 as a therapeutic agent.

14. Use of the liposome or gel according to claim 13, wherein, The liposome or gel contains an antibacterial drug.

15. Use of the liposome or gel according to claim 13 or 14, wherein, The use is for treating bacterial infections, viral infections, protozoal infections or fungal infections.

16. Use of the liposome or gel according to claim 15, wherein, The bacterial infection is a biofilm-related bacterial infection.

17. Use of the liposome or gel according to claim 15 or 16, wherein, The bacterial infection is a bacterial infection caused by Escherichia coli or Staphylococcus aureus.

18. Use of the liposome or gel according to claim 13, wherein, The use is for treating cancer and / or preventing or inhibiting cancer metastasis.

19. Use of the liposome or gel according to claim 18, wherein, The treatment of cancer is to inhibit the growth of the primary tumor.

20. Use of the liposome or gel according to claim 18 or 19, wherein, The liposome or gel contains an anticancer drug.

21. Use of the liposome or gel according to any one of claims 18 - 20, wherein, The cancer is selected from the following list: Breast cancer, head and neck cancer, gastric cancer, prostate cancer, non-small cell lung cancer, leukemia, skin cancer, melanoma, liver cancer, ovarian cancer, pancreatic cancer, lung cancer, kidney cancer, glioma or glioblastoma in brain cancer.

22. A pharmaceutical composition comprising the liposome according to any one of claims 1-9 or the gel according to any one of claims 10-11, and at least one pharmaceutically acceptable excipient and / or adjuvant.

23. The pharmaceutical composition according to the preceding claim, wherein, The pharmaceutically acceptable excipients are selected from the following list: Diluents, buffers, preservatives, stabilizers, surfactants, solvents, disintegrants, antioxidants, binders, lubricants and colorants.

24. The pharmaceutical composition according to the preceding claim, wherein, The stabilizer or surfactant is an anionic lipid, and / or Calculated by weight percentage, the concentration of the stabilizer and surfactant in the composition is 5-45% relative to the total weight of the pharmaceutical composition.

25. The pharmaceutical composition according to any one of claims 22-24, wherein, The pharmaceutical composition is selected from the following list: Topical compositions, injections and oral compositions.

26. The pharmaceutical composition according to any one of claims 22-25, wherein, The pharmaceutical composition is a vaccine.

27. Use of the pharmaceutical composition according to any one of claims 22-25 as a therapeutic agent.

28. Use of the pharmaceutical composition according to claim 27, wherein, The liposome or gel in the pharmaceutical composition contains an anticancer drug.

29. Use of the pharmaceutical composition according to claim 27 or 28, wherein, The use is for treating or preventing cancer.

30. Use of the pharmaceutical composition according to the preceding claims, wherein, The use is to inhibit the growth of the primary tumor or shrink the primary tumor.

31. Use of the pharmaceutical composition according to claim 29, wherein, The use is to prevent or reduce cancer metastasis.

32. Use of the pharmaceutical composition according to any one of claims 28 - 31, wherein, The cancer is selected from the following list: colorectal cancer, lung cancer, pancreatic cancer and gastric cancer; or For treating a combination of cancers selected from the following cancers: colorectal cancer, lung cancer, gastric cancer, breast cancer, head and neck cancer, prostate cancer, non-small cell lung cancer, leukemia, skin cancer, liver cancer, ovarian cancer, pancreatic cancer, lung cancer and kidney cancer.

33. A method for preparing the liposome according to any one of claims 1-9, comprising at least the following steps: (i) Providing a mixture, the mixture comprising: An aqueous buffer; At least one anticancer drug, antibacterial drug or other drug; and The glyceride of the general formula (I); Wherein, the concentration of the glyceride of the general formula (I) in the mixture is equal to or lower than 3 mg / ml; preferably 0.04 mg / ml to 3 mg / ml; (ii) Stirring the mixture at a temperature higher than 40 °C, Wherein, when the anti-cancer drug, antibacterial drug or other drug is a hydrophobic compound, the anti-cancer drug, antibacterial drug or other drug is added to a mixture containing glycerides of general formula (I) obtained by prior preparation, and when the anti-cancer drug, antibacterial drug or other drug is a hydrophilic compound, the anti-cancer drug is added to the aqueous buffer solution.

34. The method according to the preceding claim, wherein The mixture in step (i) does not contain phospholipids.

35. The method according to any one of claims 33 or 34, wherein The hydrophobic anti-cancer drug, hydrophobic antibacterial drug or other drug in step (i) has been pre-dried before mixing with the buffer solution.

36. The method according to claim 34, wherein The hydrophobic anti-cancer drug, hydrophobic antibacterial drug or other drug in step (i) forms a thin film through the pre-drying.

37. The method according to any one of claims 33-36, further comprising: Step (iii), step (iii) includes: extruding the mixture obtained in step (ii), preferably extruding through a membrane, or by sonication.

38. The method according to any one of claims 33-37, wherein, In step (i), at least one anti-cancer drug is added.

39. A method for preparing a gel as claimed in any one of claims 10-11, comprising at least the following steps: (i) Providing a mixture, the mixture comprising: An aqueous buffer solution; At least one anti-cancer drug, an antibacterial drug or other drug; and Glycerides of general formula (I); Wherein the concentration of the glycerides of general formula (I) in the mixture is higher than 3 mg / ml; (ii) Stirring the mixture at a temperature higher than 40 °C, Wherein, when the anti-cancer drug, antibacterial drug or other drug is a hydrophobic compound, the anti-cancer drug, antibacterial drug or other drug is added to the mixture containing the glycerides of general formula (I), and when the anti-cancer drug, antibacterial drug or other drug is a hydrophilic compound, the anti-cancer drug, antibacterial drug or other drug is added to the aqueous buffer solution.

40. The method according to claim 39, wherein, At least one anti-cancer drug is added in step (i).

41. The method according to any one of claims 33-40, wherein, The aqueous buffer solution is selected from the following list: Phosphate buffered saline, sodium chloride buffer solution, sterile water and combinations thereof.

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