Long-acting analgesic

Abstract

The present invention relates to a composition comprising a β-cyclodextrin polymer (a) and at least one compound (b) selected from mycolactones and their derivatives.

Description

[Technical Field]

[0001] The present invention relates to a composition comprising a β-cyclodextrin polymer and at least one compound selected from mycolactones and their derivatives, and in particular to its use for the treatment of pain. [Background technology]

[0002] Globally, more than 100 million people suffer from chronic pain associated with numerous conditions (osteoarthritis, cancer, lower back pain, sciatica, shingles, neuropathic pain, burns). Neuropathic pain, in particular, resulting from direct damage to sensory neurons, is resistant to most treatments. Pain is one of the most difficult and unbearable symptoms for patients to treat. Currently, many treatments are available to alleviate pain, particularly those based on the use of morphine derivatives. These treatments have many limitations, including short duration of action, narrow therapeutic range, serious side effects (nausea, constipation, balance disturbances, drowsiness, addiction), and an increased risk of dependence in patients. Therefore, finding alternatives that provide effective and sustained pain relief is truly needed.

[0003] The use of mycolactones and their derivatives as analgesics appears to be a promising alternative. Indeed, International Publication No. 2015 / 189342 describes the use of these molecules for the treatment of pain, as well as their mechanisms of action as angiotensin II type 2 receptor (AT2R) receptor agonists, type I cyclooxygenases, and potassium channel agonists of the TRAAK family. This involves neuronal hyperpolarization, interfering with the transmission of pain-causing nerve signals to the brain. Therefore, this use has the advantage of not inducing patient dependence.

[0004] Nevertheless, the use of this potent analgesic is greatly limited by the chemical properties of mycolactone derivatives, making their administration difficult. These polyketide derivatives are highly sensitive to light and therefore decompose rapidly. Furthermore, their chemical properties make them completely insoluble in aqueous media, requiring the use of organic solvents for administration. Finally, once administered, mycolactones and their derivatives have poor diffusivity in the body.

[0005] The use of nanoparticles or polymers to facilitate the administration of particularly poorly soluble drugs is currently a significant area of ​​medical research. Gouveia et al., Nature Scientific Reports 7,5390 (2017), describe the use of β-cyclodextrin nanoparticles to encapsulate ethionamide (ETH), which is used in the treatment of tuberculosis. The incorporation of the active ingredient also preserves its activity, improves the release of the drug into the lungs, and thus makes the treatment of tuberculosis more effective. However, this formulation has only been tested for administration to the lungs.

[0006] Therefore, there is a need to provide new and more effective treatments for pain.

[0007] Furthermore, there is a need to find suitable compositions that protect mycolactones and their derivatives from degradation, avoiding their potential toxic effects, while enabling the slow and sustained release of mycolactones and their derivatives.

[0008] This invention particularly satisfies these needs.

[0009] Indeed, as demonstrated in the examples, the inventors have surprisingly found that β-cyclodex We discovered that a combination of a trin polymer and a molecule selected from mycolactone or its derivatives enables the solubilization of mycolactone or its derivatives in an aqueous medium without the use of organic solvents. Furthermore, it advantageously protects the mycolactone from degradation under UV irradiation. [Overview of the project]

[0010] Therefore, the first object of the present invention is -(a)β-cyclodextrin polymer and -(b) A composition comprising at least one compound selected from mycolactone and its derivatives.

[0011] A second object of the present invention is a method for preparing the composition.

[0012] Another object of the present invention is a pharmaceutical composition comprising at least the composition according to the present invention and at least one pharmaceutically acceptable excipient.

[0013] Another object of the present invention is a composition for use in the treatment of pain.

[0014] The composition according to the present invention comprises compound (a), which is a β-cyclodextrin polymer.

[0015] The term "β-cyclodextrin polymer" refers to all compounds that can be obtained by polymerization of compounds selected from the β-cyclodextrin family, or by grafting compounds selected from the β-cyclodextrin family onto a (co)polymer.

[0016] The term "cyclodextrin" has a common meaning in the prior art and refers to a family of cyclic oligosaccharides consisting of macrocyclic compounds containing several glucopyranose subunits linked together by 1,4-glycosidic bonds.

[0017] Cyclodextrins are mainly produced by the enzymatic degradation of amylose and its derivatives.

[0018] The cyclodextrins mainly used are classified into three families: α-, β-, and γ-cyclodextrins. These families are distinguished by the number of units constituting the cyclic structure of cyclodextrin. "α-Cyclodextrin" is composed of six glucopyranose groups in its cyclic structure, "β-cyclodextrin" is composed of seven glucopyranose groups in its cyclic structure, and "γ-cyclodextrin" is composed of eight glucopyranose groups in its cyclic structure.

[0019] The chemical structure of β-cyclodextrin is as shown in formula (A).

[0020] [Chemical formula] The β-cyclodextrin molecule has a frustum of a cone structure and forms a cavity at its center. This cavity has a non-polar and hydrophobic carbon environment (carbon skeleton and oxygen in ether bonds) that can accommodate poorly water-soluble molecules. On the other hand, the outer side of the frustum of the cone has a large number of hydroxyl groups, resulting in good solubility of β-cyclodextrin in an aqueous medium. This amphiphilic property also enables β-cyclodextrin to encapsulate hydrophobic molecules (or parts thereof) in its cavity and form an inclusion complex in an aqueous medium.

[0021] The formation of these inclusion complexes enables the encapsulation of various molecules, particularly mycolactone and possibly other molecules for therapeutic purposes, within the β-cyclodextrin molecule. Advantageously, the cyclodextrin polymer contains more than 20 β-cyclodextrin linkages, but not all of them are occupied by mycolactone. This allows the simultaneous incorporation of active molecules for various purposes in families such as antibiotics, analgesics, and anti-cancer drugs.

[0022] The term "inclusion complex" refers to a system composed of a host molecule (in this case, β-cyclodextrin) that can accommodate chemical species (in this case, mycolactone and its derivatives).

[0023] The term "encapsulating" indicates that a chemical species is included within a host molecule.

[0024] According to one embodiment, the β-cyclodextrin polymer (a) is prepared by a polymerization reaction between a modified or unmodified β-cyclodextrin and at least one other chemical compound selected from the group consisting of epichlorohydrin, tartaric acid, citric acid, acylated poly(ethylene glycol), adipoyl chloride, succinyl chloride, glutaraldehyde, diphenyl carbonate, 1,4-butanediol diglycidyl ether, toluene diisocyanate, naphthalene diisocyanate, succinic anhydride, 1,2,4,5 benzene tetracarboxylic anhydride, monochlorotriazine, trimethoxysilane derivative, diethynylbenzene, and tetrafluoroterephthalonitrile. Another type of cyclodextrin polymer is obtained from cyclodextrin-based organic-inorganic porous particles (metal-organic frameworks, MOFs) and can then be crosslinked using the agents described above in this paragraph. Alternatively, a water-soluble copolymer can be grafted onto cyclodextrin. These copolymers are among the group consisting of alginate, poly(ethyleneimine), poly(N-hydroxyethylacrylamide), chitosan, hyaluronic acid, phenylalanine, polyanhydride, polyaspartoamide, and cellulose.

[0025] According to another embodiment, the polymer (a) is a reaction product between a modified or unmodified β-cyclodextrin and epichlorohydrin.

[0026] According to another embodiment, the polymer (a) is a reaction product between a modified β-cyclodextrin and epichlorohydrin.

[0027] According to another embodiment, the polymer (a) is a reaction product between a modified β-cyclodextrin and epichlorohydrin.

[0028] "Modified β-cyclodextrin" refers to any β-cyclodextrin that has been modified by at least one chemical reaction without altering the inherent properties of the β-cyclodextrin.

[0029] Chemical reactions, in particular, refer to addition reactions, substitution reactions, or polymer graft reactions.

[0030] Generally, the modified β-cyclodextrins used in accordance with the present invention are β-cyclodextrins in which their hydroxyl groups, preferably the secondary hydroxyl groups of each glucose unit forming them, are modified by the addition of one or more identical or different substituents.

[0031] The "modified β-cyclodextrin" used in the composition according to the present invention is a β-cyclodextrin having one or more identical or different substituents selected from radicals containing functionalized or unfunctionalized alkyl radicals, hydroxyalkyl radicals, carboxyl radicals, carboxylates, nitro, amino, sulfonates, sulfates, phosphates, ethers, polyethers, ammonium, and ester functional groups. The alkyl radical may be a linear or branched alkyl radical having 1 to 20 carbon atoms, preferably 1 to 5 carbon atoms, more specifically a methyl or ethyl radical. Examples of hydroxyalkyl radicals include hydroxyalkyl groups having 1 to 20 carbon atoms, preferably 1 to 5 carbon atoms, more specifically -CH2CH2OH, or -CH2OH. Examples of radicals containing ester functional groups include ester groups having 1 to 20 carbon atoms, preferably 1 to 5 carbon atoms, more specifically the groups -O(CO)CH3 and -O(CO)CH2CH3.

[0032] Examples include methylated, ethylated, propylated, succinylated, carboxylated, acetylated, 2-hydroxypropylated, and polyoxyethylated cyclodextrins.

[0033] Preferably, polymer (a) is a crosslinked polymer, i.e., the polymer has a three-dimensional nonlinear network structure formed by the creation of bonds between polymer chains during the polymerization reaction. This polymer can then be modified by adding new functional groups, such as negative or positive charges, or by grafting fluorescent molecules.

[0034] Preferably, polymer (a) is a reaction product between β-cyclodextrin and epichlorohydrin.

[0035] According to this embodiment, this polymer is known and is referred to as pCD in Gref et al. Journal of Controlled Release: Official Journal of the Controlled Release So This is described in Ciety 2006, 111(3), 316-324.

[0036] Polymer (a) can be prepared by adapting or applying the methods described in Gref et al., Journal of Controlled Release: Official Journal of the Controlled Release Society 2006, 111(3), 316-324, and Othman et al., J Colloid Interface Sci 2011, 354(2), 517-27.

[0037] More specifically, polymer (a) can be prepared by a polymerization reaction between β-cyclodextrin and epichlorohydrin in an alkaline medium in the presence of NaOH. For example, β-cyclodextrin is dissolved in a solution containing 33% by mass of NaOH with vigorous stirring, and then epichlorohydrin is added with stirring, stopping the reaction by adding acetone before the gelation point.

[0038] According to this production method, the chemical structure of polymer (a) is represented by general formula (B):

[0039] [ka] In the formula, the frustum of a cone represents a β-cyclodextrin molecule, and n corresponds to the reacted epichlorohydrin molecule, representing the repeating unit separating the two β-cyclodextrin molecules.

[0040] The composition according to the present invention comprises compound (b) selected from mycolactone and its derivatives.

[0041] Mycolactones and their derivatives are 12-membered polyketide derivatives originally produced by various bacterial strains of the Mycobacterium ulcerans (Mu) family, which are involved in Buruli ulcer (BU).

[0042] Compound (b) is readily available.

[0043] These can be obtained, for example, by extraction and purification from Mycobacterium ulcerans extract. This method is described in George et al.; SCIENCE Volume 283, Issue 5403, 5 February 1999.

[0044] It is possible to use different strains of Mycobacterium ulcerans that produce different isomers. Isomer production varies depending on the region from which the bacterial strain originates.

[0045] The mycolactones used in these examples are derived from strain 1615, which corresponds to the bibliographic reference cited in George et al; SCIENCE Volume 283, Issue 5403, 5 February 1999. It is also possible to extract mycolactones from strains obtained directly from patients.

[0046] Alternatively, the compounds can be synthesized. A synthetic method for preparing mycolactones and their derivatives has been developed and is described in European Patent No. 2 594 561.

[0047] According to one embodiment, compound (b) is a compound of formula (I),

[0048] [ka] During the ceremony, -R1, R2, R4, and R5 are identical or different, and are independently selected from the group consisting of H, R6, C(O)R7, C(S)R7, C(O)NHR7, and C(S)NHR7. -R3 is selected from the group consisting of H, OH, OR6, OC(O)R7, OC(S)R7, OC(O)NHR7, OC(S)NHR7, and OCH(OH)R7. -R6 is C1-C6 alkyl, C6-C 12 Ariel, C6-C 12 A group selected from the group consisting of heteroaryls and sugar derivatives, -R7 is H, C1-C6 alkyl, C6-C 12 Heteroaryls, and C6-C 12 Selected from the group of aryls, here, -R1 and R2, R2 and R3, and / or R4 and R5 together form an acetal group in the compound. and its pharmaceutically acceptable salts.

[0049] The term "C1-C6 alkyl" group, unless otherwise specified, refers to a linear or branched aliphatic hydrocarbon group containing a total of 1 to 6 carbon atoms and potentially containing one or more unsaturated atoms. Examples include methyl, ethyl, n-propyl, butyl, isobutyl, tert-butyl, pentyl, and hexyl groups.

[0050] "C6-C 12The term "aryl" group, unless otherwise specified, refers to monocyclic, bicyclic, or tricyclic aromatic hydrocarbon compounds containing a total of 6 to 12 carbon atoms. Examples include phenyl and naphthyl groups.

[0051] "C6-C 12 The term "heteroaryl" group refers to a group containing a total of 6 to 12 carbon atoms. The term "monocyclic, bicyclic, or tricyclic aromatic compound" refers to one of the monocyclic, bicyclic, or tricyclic aromatic compounds in which at least one atom is a heteroatom selected from the group consisting of nitrogen, phosphorus, oxygen, and sulfur. Examples include pyrrole, furan, thiophene, imidazole, furazan, oxazole, oxadiazole, oxatriazole, isoxazole, thiazole, isothiazole, pyrazole, triazole, and tetrazole groups. The term "sugar derivative" refers to a compound containing at least one sugar unit selected from monosaccharides and polysaccharides. Examples of monosaccharide units include glucose, galactose, fructose, and pentose.

[0052] According to another embodiment, compound b) is compound (II),

[0053] [ka] and its pharmaceutically acceptable salts.

[0054] This compound is a compound of formula (I), in which R1, R2, R4, and R5 are -H groups, and R3 is an -OH group, and corresponds to a mycolactone.

[0055] According to one embodiment, the composition according to the present invention comprises a compound (b) encapsulated in a β-cyclodextrin polymer (a). Encapsulation in polymer (a) means the incorporation of mycolactone by solubilization in an aqueous medium. In practice, this is achieved by adding an aqueous solution of the β-cyclodextrin polymer (a) to a container containing pure mycolactone. Upon contact with (a), the mycolactone is solubilized with or without mechanical agitation. Alternatively, sonication or Ultraturrax may be used to accelerate the process.

[0056] While we do not wish to be bound by any particular theory, the mycolactone and / or derivative molecules form inclusion complexes with the β-cyclodextrin molecules constituting polymer (a). The formation of these inclusion complexes also allows compound (b) to be encapsulated within the β-cyclodextrin molecules constituting the polymer.

[0057] Advantageously, the formation of these inclusion complexes allows for the spontaneous solubilization of compound (b) without the use of organic solvents.

[0058] Advantageously, encapsulation of compound (b) in β-cyclodextrin polymer (a) also significantly stabilizes compound (b) against degradation, particularly under UV irradiation.

[0059] Advantageously, mycolactones and their derivatives have a very strong affinity for β-cyclodextrins. The affinity allows for the sustained release of compound (b) into the body after administration of the composition.

[0060] In other words, the composition according to the present invention is a composition comprising a β-cyclodextrin polymer in which compound (b) is encapsulated.

[0061] Preferably, the composition according to the present invention may be in powder form or an aqueous solution, and more preferably, the composition is an aqueous solution.

[0062] In particular, the powder can be obtained by freeze-drying the composition according to the present invention in an aqueous solution.

[0063] Advantageously, encapsulation of compound (b) with β-cyclodextrin polymer makes it possible to obtain aqueous compositions containing high concentrations of compound (b).

[0064] According to one embodiment, the composition is an aqueous solution, and the concentration of compound (b) is 0.001 mg / mL to 10 mg / mL, preferably 0.05 to 5 mg / mL.

[0065] Preferably, the composition according to the present invention is an aqueous solution, and the concentration of compound (a) is 25 mg / mL to 250 mg / mL, preferably 50 mg / mL to 150 mg / mL.

[0066] Preferably, the composition according to the present invention includes a ratio of the amount of compound (b) by weight to the amount of polymer (a) by weight, which can be 0.1 to 10, preferably 0.5 to 7, and more preferably 1 to 5 (the percentages are expressed relative to the total weight of the composition).

[0067] According to another embodiment, the β-cyclodextrin polymer (a) can typically encapsulate at least one other active ingredient having a complementary, enhancing, or synergistic effect with compound (b), for example. Generally, the active ingredient is encapsulated within the cavities of free (i.e., not yet forming an inclusion complex with compound (b)) β-cyclodextrin molecules that form polymer (a).

[0068] According to one embodiment, the composition according to the present invention may further comprise one or more active components (c) selected from anticancer agents, analgesics, anesthetics, active molecules having a healing effect, antibacterial agents, antibiotics, bactericidal agents, hemostatic agents, antifungal agents, antiviral agents, antithrombotic agents, anti-inflammatory agents, antipruritic agents, contrast agents, and hormones.

[0069] Typically, the active ingredient is an antimicrobial agent such as chlorhexidine or benzalkonium chloride, ethanol, hexamidine, betadine, chlorinated derivatives, or triclocarban.

[0070] In another embodiment, the active ingredient (c) is an analgesic such as lidocaine, codeine, or tramadol.

[0071] In another embodiment, the active ingredient (c) is an antibiotic such as vancomycin (VCM), amikacin, gentamicin, or amoxicillin.

[0072] In another embodiment, the active ingredient (c) is an anticancer agent such as doxorubicin, cyclophosphamide, cisplatin, docetaxel, gemcitabine, or oxaliplatin.

[0073] Compound (b) and the active ingredient may be available in the form of a hydrate and / or a pharmaceutically acceptable salt. These compounds may exist. In fact, they may exist in the form of corresponding salts of pharmaceutically acceptable organic or inorganic acids or organic or inorganic bases.

[0074] The term "pharmaceutically acceptable salts" refers to relatively non-toxic inorganic and organic acid addition salts, as well as base addition salts, of the compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds. In particular, acid addition salts can be prepared by reacting the purified compound separately with an organic or inorganic acid in its purified form and isolating the salts thus formed. Examples of acid addition salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptanoate, lactobionate, sulfamate, malonate, salicylate, propionate, methylenebis-β-hydroxynaphthate, gentisic acid, isethionate, di-p-toluyl tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfonate and quinate, laurylsulfonate, and their analogues. (See, for example, SMBerge et al. "Pharmaceutical Salts" J.Pharm.Sci, 66:p.1-19 (1977)). Acid addition salts can also be prepared by reacting a purified compound separately with an organic or inorganic base in its acidic form and isolating the salt thus formed. Acid addition salts include amino and metal salts. Preferred metal salts include sodium, potassium, calcium, barium, zinc, magnesium, and aluminum salts. Sodium and potassium salts are preferred. Preferred inorganic base addition salts are prepared from metal bases including sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, and zinc hydroxide.Suitable basic amino addition salts are prepared from amines having sufficient alkalinity to form stable salts, and preferably include amines often used in medicinal chemistry due to their low toxicity and tolerance for medical applications: ammonia, ethylenediamine, N-methylglucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabiethylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids (e.g., lysine and arginine), and dicyclohexylamine, as well as their analogues.

[0075] According to another embodiment, the β-cyclodextrin polymer (a) can be advantageously combined with other compounds such as lipid compounds (injectable oils such as cholesterol and Migliol), glycoproteins (e.g., lamins), and nanoparticles (nanoparticles, NPs), thereby advantageously imparting complementary properties to the composition according to the present invention, such as antibacterial or antifungal properties or targeting properties, or allowing other active ingredients to be included in the composition.

[0076] This encapsulation is generally feasible if the size of the nanoparticles allows them to be contained within the cavities of free β-cyclodextrin molecules.

[0077] Preferably, the size of the nanoparticles (d) is 50 nm to 400 nm, more preferably 50 nm to 250 nm. The lipid compound can form microdroplets in the formulation ranging in size from 50 nm to 2 to 5 microns.

[0078] For another purpose, the present invention also relates to a composition according to the present invention and at least one pharmaceutically This relates to a pharmaceutical composition comprising acceptable excipients.

[0079] The pharmaceutical compositions of the present invention may be provided in various forms intended for topical, parenteral, or intrathecal administration. Preferably, the compositions are administered to patients who require them.

[0080] Local administration refers to the application of a product to the surface of the body, such as the skin or mucous membranes.

[0081] Parenteral administration refers to the administration of a product by injection. Parenteral administration includes intramuscular (IM), intravenous (IV), subcutaneous (SC), intradermal, intra-arterial, or intra-articular injections.

[0082] Intrathecal administration refers to injecting the drug into the subarachnoid space, allowing it to reach the cerebrospinal fluid, and then dispersing the injected product.

[0083] In particular, injectable pharmaceutical forms for parenteral or intrathecal use are generally injectable solutions that can be obtained by, for example, the following process: the composition according to the present invention is dissolved, suspended, or emulsified in an aqueous medium (e.g., distilled water, physiological saline, or Ringer's solution) together with a dispersant (e.g., Tween® 80, HCO® 60 (Nikko Chemicals), polyethylene glycol, carboxymethylcellulose, or sodium alginate), a preservative (e.g., methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, benzyl alcohol, chlorobutanol, or phenol), an isotonic agent (e.g., sodium chloride, glycerol, sorbitol, or glucose), and optionally other additives, such as, if desired, a solubilizer (e.g., sodium salicylate or sodium acetate), or a stabilizer (e.g., human serum albumin).

[0084] Pharmaceutical forms for topical use can be obtained from solid, semi-solid, or liquid compositions containing the compositions of the present invention. For example, to obtain a solid form, the compositions according to the present invention may be mixed with excipients (e.g., lactose, trehalose, sucrose, mannitol, starch, microcrystalline cellulose, or sucrose) and thickeners (e.g., natural rubber, cellulose derivatives, or acrylic polymers) to convert them into powders. Liquid pharmaceutical compositions are prepared in substantially the same manner as those for injectable forms, as described above. Semi-solid pharmaceutical forms are preferably in the form of aqueous or oily gels or ointments. These compositions may optionally contain pH adjusters (e.g., carbonic acid, phosphoric acid, citric acid, hydrochloric acid, or sodium hydroxide), antioxidant compounds (vitamin C, tocopherol, polyphenols), and / or preservatives (e.g., p-hydroxybenzoate, chlorobutanol, or benzalkonium chloride).

[0085] According to one embodiment, the pharmaceutical composition according to the present invention comprises a pharmaceutically acceptable excipient selected from dextran and its derivatives (preferably hydrophobized dextran).

[0086] Advantageously, the use of hydrophobized dextran in the pharmaceutical composition according to the present invention enables the formation of gels or nanoparticles. In this case, it is possible to incorporate nanoparticles or oil droplets, as described in Example 13.

[0087] Preferably, the size of the nanoparticles (d) is 50 nm to 400 nm, more preferably 50 nm to 250 nm. The lipid compound can form microdroplets in the formulation ranging in size from 50 nm to 2 to 5 microns.

[0088] According to one embodiment, the composition according to the present invention comprises inorganic nanoparticles and organic-inorganic hybrids. The material may further include nanoparticles (d) selected from nanoparticles or polymer nanoparticles.

[0089] The term "nanoparticles" (NPs) generally refers to spherical particles having an average diameter (number mean diameter) of 1 nm to 1 μm. In the context of the present invention, these NPs typically have an average diameter of less than 400 nm, particularly less than 250 nm, and a polydispersity index of less than 0.25.

[0090] The average diameter (number mean diameter) and polydispersity can be measured by dynamic light scattering (DLS) or transmission electron microscopy (TEM).

[0091] Examples of inorganic nanoparticles include magnetic nanoparticles, silver nanoparticles with antibacterial properties, gold nanoparticles, and copper nanoparticles.

[0092] Organic-inorganic hybrid nanoparticles include MIL-100(Fe) iron trimetate particles, other iron carboxylates, and UIO-66.

[0093] Examples of polymer nanoparticles include PLGA ((poly(lactic-co-glycolic acid))), poly(ethylene glycol)(PEG)-PLGA (poly(ethylene glycol)(PEG)-PLGA, PEG-PLGA) copolymer, and polycaprolactone (PCL) particles.

[0094] Advantageously, the nanoparticle(d) may itself contain an active ingredient.

[0095] To another purpose, the present invention also relates to a method for preparing a composition according to the present invention, comprising the step of mixing compounds (a) and (b) in an aqueous solution.

[0096] According to another embodiment, the process according to the present invention may optionally include the step of adding an active ingredient (c) and / or nanoparticles (d) to an aqueous solution.

[0097] Preferably, the process according to the present invention may also include a freeze-drying step to obtain the composition in powder form.

[0098] Another object of the present invention relates to the use of the compositions according to the present invention for the treatment of pain.

[0099] The term “pain” also refers to “painful disorders,” which describe suffering caused by trauma, burns, lesions, or any other cause, to varying degrees in intensity, and which themselves manifest as a disruption of well-being, a disruption of health balance, or a loss or reduction of physical integrity.

[0100] Pain is generally classified into acute or chronic. "Acute" pain refers to a sudden, short-term experience associated with a specific cause, such as certain trauma resulting from surgery, dental treatment, or a sprain.

[0101] "Chronic pain" refers to long-term pain that causes significant psychological and emotional problems. This type of pain is associated with many conditions, including osteoarthritis, cancer, and diabetes.

[0102] The compositions according to the present invention are suitable for the treatment of a wide range of pain disorders, including, when these manifest as symptoms of a disease, particularly acute pain, chronic pain, neuropathic pain, inflammatory pain, iatrogenic pain (including cancer pain), infectious pain (including herpes pain), visceral pain, central pain, dysfunctional pain (including fibromyalgia), and nociceptive pain (including postoperative pain). This also includes mixed-type pain (including cancer pain, back pain, and orofacial pain, involving the visceral, gastrointestinal, cranial, musculoskeletal, spinal, genitourinary, cardiovascular, and CNS systems).

[0103] In general, pain disorders are considered symptoms of a medical condition that must be treated independently of the condition itself, for the sake of the patient's well-being.

[0104] The term "symptoms" refers to abnormal findings caused by the medical condition that a patient complains of.

[0105] As used herein, the term “pain management” refers to the reduction, slowing of progression, or elimination of painful disorders as described herein, and does not necessarily require treatment of the underlying conditions that may cause them. Therefore, pain management can typically be symptomatic and possibly etiological.

[0106] Generally, pain or "pain disorders" in patients are usually treated by administering short-acting analgesics or painkillers.

[0107] The term "analgesic" refers to drugs such as opioid derivatives that are used to treat pain. This refers to drugs used to suppress sexual desire.

[0108] "Analgesics" refer to drugs used to reduce pain.

[0109] Preferably, the composition according to the present invention is used as an analgesic for treating pain or painful disorders.

[0110] According to another embodiment, the composition according to the present invention is used as an analgesic for treating pain and painful disorders.

[0111] Advantageously, the compositions according to the present invention can be used in a complementary manner to treat painful disorders caused by medical conditions. [Brief explanation of the drawing]

[0112] [Figure 1] A: Improved mycolactone (ML) inclusion due to increased pCD volume. B: Sequential extraction of ML from ML-pCD. [Figure 2] The protective role of pCD against the degradation of mycolactones exposed to UV light. [Figure 3]A) There are no additional cytotoxic effects caused by pCD. B-D) pCD does not alter the immunomodulatory effects of mycolactones (production of IL-6, TNF-α, and IL-1β). Black = Free ML, Gray = Integrated ML (pCD-ML) [Figure 4] Analgesic effect of mycolactone encapsulated in pCD in mice (#: difference between ML and pCD-ML). [Figure 5] Biodegradable cyclodextrins reduce the analgesic effect of mycolactones compared to pCD. [Figure 6] Analgesic effect of mycolactone encapsulated in pCD in a mouse plantar incision model (surgical pain model) (#: difference between ML and pCD-ML). [Figure 7] Analgesic effect of mycolactone encapsulated in pCD in a mouse plantar incision model (surgical pain model) (#: difference between ML and pCD-ML). [Examples]

[0113] Example 1 - Synthesis and characterization of cyclodextrin polymer (pCD) In a 500 mL flask, add 50 g of β-CD to 80 mL of 33% NaOH and stir overnight. The next day, heat the flask to 30°C and add 35 mL of epichlorous acid. Add rohydrin (EP) while stirring (Scheme 1). Stir the reaction mixture at 30°C for 1 to 1.5 hours to ensure complete homogenization. If viscosity increases, stop the reaction by adding 80 mL of acetone just before the freezing point.

[0114] [ka] Figure 1. Reaction diagram of the synthesis of pCD from β-CD crosslinked with epichlorohydrin.

[0115] The bath temperature is raised to 50°C and the mixture is stirred overnight. The next day, it is neutralized with a 6M HCl solution to obtain a pH of 7. Insoluble residue is removed, and the remaining polymer is purified by dialyzing using a Spectra / por membrane (cutoff threshold 100,000 g / mol) and then freeze-dried. The resulting pCD polymer is then characterized by NMR and SEC to determine its β-CD content. The β-CD content is approximately 70% (by weight).

[0116] Using the same methodology, α-CD polymers and γ-CD polymers are synthesized.

[0117] pCD samples can be fractionated by continuous dialysis (e.g., using SpectraPor membranes with cutoff thresholds of 20, 50, 100, 300, and 1000 kDa) to obtain samples with low polydispersity.

[0118] Example 2 - Synthesis and Characterization of Malic Acid-Based Cyclodextrin Polymers In a 25 mL flask, 0.2 mmol of NaH2PO4·2H2O (catalyst), 0.09 mmol of β-CD, and 0.45 mmol of malic acid are mixed in 2 mL of water. The mixture is then concentrated by evaporation at 140°C for 10 minutes, and then heated under reduced pressure (vacuum pump) at 140°C for 25 minutes. The polymer thus formed is dissolved in 10 mL of Milli-Q water and dispersed by sonication. The insoluble fraction is removed by filtration. The soluble fraction is then purified by dialysis (Spectra / por membrane, cutoff threshold 20,000 g / mol), and then freeze-dried. The polymer is then ( 1)H The β-CD content is determined by characterization using NMR and SEC. The β-CD content is approximately 70% (by weight).

[0119] This polymer is called pCD.

[0120] Biodegradable β-CD polymers are synthesized in a similar manner, by replacing malic acid with citric acid.

[0121] Example 3 - Synthesis of alkyl chain-modified dextran To synthesize dextran grafted with hydrophobic lauryl chains (DM), 4 g of dextran (40,000 g / mol) was dissolved in 100 mL of dimethylformamide containing 1 g of lithium chloride. Next, 0.62 mL of lauryl chloride and 0.031 mL of pyridine were added to the dextran solution. The reaction was carried out at 80°C for 3 hours. The resulting MD was isolated by precipitation in isopropyl alcohol. It was then purified by dissolving in distilled water and dialyzing for 48 hours, and subsequently lyophilized. 1)H The grafting rate of alkyl chains, as determined by NMR, is approximately 6%.

[0122] Example 4 - Mycolactone Production Mycolactones are purified from a culture of M. ulcerans strain 1615 according to the protocol described in Georges 1999(1). Briefly, the total lipids of mycobacteria are extracted using the Folch method. Then, the phospholipids are precipitated in cold acetone, and the supernatant containing mycolactones is deposited on a silica plate for thin-layer chromatography. After development, the silica with adsorbed mycolactones (rf value 0.23) is scraped off, and the mycolactones are desorbed from the silica by filtration (in chloroform / methanol). Finally, the mycolactones are quantified by high-performance liquid chromatography (2). 1. George KM, Chatterjee D, Gunawardana G, Welty D, Hayman J, Lee R, Small PL. 1999. Mycolactone: a polyketide toxin from Mycobacterium ulcerans required for virulence. Science 283:854-857. 2. Marion E, Prado S, Cano C, Babonneau J, Ghamrawi S, Marsollier L. 2012. Photodegradation of the Mycobacterium ulcerans toxin, mycolactones: considerations for handling and storage. PLoS One 7:e33600.

[0123] Example 5 - Incorporation of mycolactone into pCD Dissolve 0.5 mg of mycolactone (ML) in ethanol and place it in a 1.5 mL amber glass vial. Evaporate the ethanol using a vacuum concentrator (SpeedVac). Then, add 1 mL of pCD solution at a concentration of 100 mg / mL. Stir the preparation for 48 hours and then store it at 4°C. This gives time for the ML to be fully incorporated into the pCD. This solution is called pCD-ML.

[0124] The experiment was repeated by replacing pCD with β-CD polymer and β-CD polymer. Even when the incubation time was extended to 72 hours, ML was not solubilized.

[0125] Example 6 - Improvement of mycolactone encapsulation by increasing the amount of pCD Starting with a 100 mg / mL pCD solution, serial dilutions were performed with MilliQ water to obtain the following pCD concentrations: 1, 10, 50, and 100 mg / mL.

[0126] 0.5 mg of ML dissolved in ethanol was introduced into a 1.5 mL amber glass vial. Evaporate the ethanol using a vacuum concentrator (SpeedVac). Then, add 1 mL of pCD solution at concentrations of 1, 10, 50, and 100 mg / mL. Shake the preparation for 48 hours.

[0127] The aliquots were collected and measured by HPLC as in Example 5 to determine the amount of ML incorporated. It was found that at least 100 mg / mL of pCD was required to incorporate 0.5 mg of ML (Figure 1A).

[0128] After incubation with stirring, 30 μL of each pCD-ML preparation is taken and diluted 1 / 10 with acetonitrile. After centrifugation (3000 g, 5 minutes), the supernatant is measured by HPLC. All pellets are collected in MilliQ water, diluted 1 / 10 with acetonitrile, centrifuged, and measured again by HPLC. These steps are repeated three times to extract all the encapsulated ML contents at the different pCD concentrations tested.

[0129] The remaining preparation is transferred to ethanol, then diluted to 1 / 10 with acetonitrile, centrifuged (3000g, 5 minutes), and measured by HPLC.

[0130] In conclusion, the higher the ML content in pCD, the more difficult it becomes to extract ML from pCD. Therefore, four consecutive extractions are required to extract all of the ML from pCD (100 mg / mL pCD and 0.5 mg of incorporated ML). This demonstrates the strong affinity of ML for pCD. At high concentrations, highly hydrophobic ML is very likely to self-associate within pCD (Figure 1B).

[0131] Example 7 - Stability of pCD-ML during storage Remarkably, the ML incorporated into the pCD no longer adsorbed onto glass or plastic containers, making them easier to handle. This was demonstrated by measuring the concentration of ML in the pCD-ML solution prepared as in Example 5 by HPLC. After 3 months, the concentration variation was less than 4%.

[0132] Example 8 - Protective role of pCD against mycolactone degradation exposed to UV light. Place 50 μL of the pCD-ML solution (corresponding to 0.5 mg / mL ML) prepared according to Example 5 into a clear glass tube. Then, expose the tube to UV irradiation (room temperature, wavelength 312 nm) for 15 minutes, 1 hour, 2 hours, 6 hours, and 24 hours. Keep one tube in the dark and use it as a control. Seal all tubes tightly to prevent evaporation.

[0133] After exposure to UV light for these different periods, ML was measured by HPLC. A series of ML extractions were performed as described in Example 6. To do this, 450 μL of acetonitrile was added to each tube containing 50 μL of pCD-ML. After 6 hours of exposure, 80% of the ML contained in ethanol was degraded, while the ML associated with pCD was only 50% degraded during the same period (Figure 2).

[0134] Example 9 - In vitro evaluation of the cytotoxic and immunomodulatory effects of pCD on mouse macrophages. Two preparations were made: an ethanol solution of ML and an aqueous solution of 0.5 mg / mL of pCD-ML. Cascade dilution was performed to obtain solutions at concentrations of 100, 10, and 1 μg / mL in ML, and then 60, 20, and 2 ng / mL. 100 μL of each dilution was dispensed per well containing 100 μL of cells (100,000 cells / well) to obtain final concentrations of ML at 30, 10, and 1 ng / mL. The cells were incubated for 24 hours (37°C, 5% CO2), and then treated with 20 μL of 500 ng / mL lipopolysaccharide (LPS) solution. The cells were stimulated, and the final concentration of LPS in each well was adjusted to 50 ng / mL. After 24 hours of incubation, the supernatant was collected.

[0135] To determine the cytotoxic effect of ML, the Lonza® ToxiLight® test (Fisher Scientific) is used. To do this, 5 μL of each supernatant is mixed with 25 μL of reagent.

[0136] The immunomodulatory effect of ML was determined using an ELISA kit (IL-6, TNF-a, IL-1b). Before use, the supernatant was diluted 1 / 10 for the IL-6 and TNF-a kits and 1 / 2 for the IL-1b kit.

[0137] The incorporation of ML into pCD does not seem to significantly alter its immunomodulatory effect (Figures 3A - 3D).

[0138] Example 10 - Analgesic Effect of Mycolactone Encapsulated in pCD in Mice An aqueous solution of pCD (100 mg / mL), an ethanol solution of ML (3 mg / mL), and a solution of pCD - ML (3 mg / mL) are prepared as described above in three different tubes. Then, pCD is diluted with saline to a concentration of 8.33 mg / mL. ML is diluted with corn oil (1) to a concentration of 250 μg / mL. Finally, the pCD - ML preparation is diluted with saline to a ML concentration of 250 μg / mL. 1. Babonneau J, Breard D, Reynaert ML, Marion E, Guilet D, Saint Andre JP, Croue A, Brodin P, Richomme P, Marsollier L. 2019. Mycolactone as Analgesic: Subcutaneous Bioavailability Parameters. Front Pharmacol 10:378.

[0139] Experiments were performed on female Balb / c mice aged 7 - 10 weeks. 15 μL of the preparation was injected subcutaneously into the plantar surface of the foot. Then, using a Hargreaves plantar test device, the latency until the mouse withdrew its foot after a thermal stimulus was measured. The values shown correspond to the ratio of the test group to the control group (pCD alone).

[0140] As shown in Figure 4, the analgesic effect induced by pCD - ML is greater than that of ML alone, and this effect persists longer. For statistical analysis, two - factor ANOVA followed by Dunnett's multiple comparison test was used: *p<0.05, ** p<0.01, **** p<0.0001).

[0141] Example 11 - Biodegradable cyclodextrin reduces the analgesic effect of mycolactone. Solutions with a concentration of 100 mg / mL were prepared from pCD and biodegradable cyclodextrin (referred to as pCD-citrate). ML was enclosed in each of these solutions at a concentration of 3 mg / mL. These preparations were then diluted to a concentration of 250 μg / mL with physiological saline. Simultaneously, a control solution without ML was prepared using the same method.

[0142] These experiments were conducted on Balb / c mice aged 7–10 weeks. 15 μL of the preparation was subcutaneously injected into the sole of the foot. Then, the latency until the mouse retracted its foot after thermal stimulation was measured using a plantar test apparatus with the Hargreaves method. The values ​​shown correspond to the ratio of the test group to the control group (pCD alone). As shown in Figure 5, the analgesic effect induced by pCD-ML was greater and longer-lasting than that of ML alone. Statistical analysis was performed using two-way ANOVA, followed by Dunnett's multiple comparison test. * p<0.05, ** p<0.01, **** p<0.0001). The inventors Under these conditions, only the incorporated ML exhibited a significant analgesic effect. This result demonstrates the specificity of ML-pCD in inducing analgesia.

[0143] Example 12 - Analgesic effect of mycolactone encapsulated in pCD in a mouse plantar incision model (surgical pain model) In two different tubes, an aqueous solution of pCD (100 mg / mL) and a solution of pCD-ML (3 mg / mL) were prepared as described above. These solutions were then diluted with physiological saline to obtain a final preparation containing 3.75 μg of ML, 1.875 μg of ML, and 0.937 μg of ML per 15 μL of solution.

[0144] The values ​​shown correspond to the ratio of the test group to the control group (pCD alone). As shown in Figures 6 and 7, the analgesic effect induced by pCD-ML was greater and longer-lasting than that of ML alone. Statistical analysis was performed using two-way ANOVA, followed by Dunnett's multiple comparison test: * p<0.05, ** p<0.01, **** p<0.0001). Under the inventors' conditions, only ML-pCD exhibits a significant analgesic effect. This result demonstrates the specificity of ML-pCD in inducing analgesia.

[0145] These experiments were conducted on 7-10 week old C57Bl / 6 mice. The flexor digitorum brevis muscle was incised. 15 μL of the preparation was subcutaneously injected as close as possible to the incision site. Then, the latency until the mouse retracted its foot after thermal stimulation was measured using a Hargreaves paw test apparatus. The results showed that a significant analgesic effect was observed for more than 30 hours after application, up to a dose of 0.9 μg of ML in pCD (Figure 7). This effect lasted longer at doses of less than 3.75 μg of ML (Figure 6A). This result is surprising, but can be explained by the fact that at low doses, the adverse effects of ML (immunomodulatory effects) that could reduce the analgesic effect are no longer present.

[0146] Example 13 - Simultaneous integration of magnetic particles Prepare DM solution and pCD solution, each at 75 mg / mL. When equal volumes of these solutions are mixed, a soft gel system is instantly formed and settles at the bottom of the container. This gel can be collected and passed through a syringe (20G needle).

[0147] The same procedure is followed, except that 0.5 mg of iron oxide nanoparticles (Aldrich, <5 microns) reference 310069 are incorporated into DM (1 mL) and incubated for 4 hours. After mixing with 1 mL of pCD, a dark gel system is formed. Because it contains magnetic particles, it can be moved using a magnet. Almost all of the magnetic particles are incorporated into this gel. Furthermore, the cohesive force is maintained even when this system is significantly diluted (by adding 3 L of water).

[0148] The same experiment is performed by incorporating ML into pCD as in Example 5, and then mixing it with a DM solution containing magnetic particles. The same type of gel is formed.

[0149] Example 14 - Development of Nanoparticles Equal volumes of DM (10 mg / mL) and pCD (10 mg / mL) are mixed, with or without ML. Nanoparticles of approximately 120 nm are instantly formed, and a turbid solution is observed.

[0150] Example 15 - Freeze-drying Aqueous solutions of pCD-ML were prepared in amber vials as described in Example 5. The final concentrations of pCD were 10 and 100 mg / mL, and the final concentrations of ML were 0.3 and 0.5 mL, respectively. These solutions were frozen directly at -80°C and then lyophilized (typically using an Alpha 1-2 LD Plus freeze-dryer at -65°C). 0.018 mbar (24 hours).

[0151] At the end of freeze-drying, a white solid is obtained. The pCD-ML solution can be easily reconstituted by adding water to this freeze-dried material.

[0152] The freeze-dried form provides better protection against photodegradation of ML than the solubilized form.

[0153] Advantageously, the lyophilized material can be stored for more than 8 months in the dark, and no degradation of the incorporated ML is detected. The integrity of the ML incorporated into the pCD was determined by HPLC (using Agilent, C18 Kinetex® 5 μm × 250 mm × 4.6 mm column, 100 Å). Detection was performed at 30°C, wavelength 363 nm, flow rate 1 mL / min, injection volume 20 μL. The gradients were as follows: 0 min = 90 / 10 (water / acetonitrile), 3 min = 50 / 50 (water / acetonitrile), 13 min = 0 / 100 (water / acetonitrile), 23 min = 0 / 100 (water / acetonitrile), 33 min = 90 / 10 (water / acetonitrile).

Claims

1. A composition, - (a) β-cyclodextrin polymer and - (b) A composition comprising at least one compound selected from mycolactones and their derivatives.

2. Compound (b) is given by the following formula (I), 【Chemistry 1】 [In the formula, -R １ , R ２ , R ４ , and R ５ may be the same or different and are independently selected from the group consisting of H, R ６ , C(O)R ７ , C(S)R ７ , C(O)NHR ７ , and C(S)NHR ７ and are selected from the group consisting of: -R ３ H, OH, OR ６ OC(O)R ７ OC(S)R ７ , OC(O)NHR ７ , OC(S)NHR ７ and OCH(OH)R ７ Selected from the group consisting of, -R ６ C １ -C ６ Alkyl, C ６ -C １２ A group selected from the group consisting of aryls and sugar derivatives, -R ７ H, C １ -C ６ Alkyl and C ６ -C １２ Selected from the group of aryls, or -R １ and R ２ , R ２ and R ３ , and / or R ４ and R ５ [They combine to form an acetal group.] The composition according to claim 1, comprising and a pharmaceutically acceptable salt thereof.

3. Compound (b) is a mycolactone corresponding to formula (II), 【Chemistry 2】 The composition according to claim 1 or 2, wherein the composition is a pharmaceutically acceptable salt thereof.

4. The composition according to any one of claims 1 to 3, wherein compound (b) is encapsulated in the β-cyclodextrin polymer (a).

5. The composition according to any one of claims 1 to 4, which exists in powder form or aqueous solution.

6. The composition according to any one of claims 1 to 5, wherein the concentration of compound (b) is 0.001 mg / mL to 10 mg / mL, preferably 0.05 to 5 mg / mL.

7. The composition according to claims 1 to 6, wherein polymer (a) is the product of a polymerization reaction between a modified or unmodified β-cyclodextrin and at least one other chemical compound selected from the group consisting of epichlorohydrin, tartaric acid, citric acid, acylated poly(ethylene glycol), adipoyl chloride, succinyl chloride, glutaraldehyde, diphenyl carbonate, 1,4-butanediol diglycidyl ether, toluene diisocyanate, naphthalene diisocyanate, succinic anhydride, 1,2,4,5-benzenetetracarboxylic anhydride, monochlorotriazine, trimethoxysilane derivatives, diethynylbenzene, and tetrafluoroterephthalonitrile.

8. The composition according to any one of claims 1 to 7, wherein polymer (a) is a polymer of modified or unmodified β-cyclodextrin crosslinked with epichlorohydrin.

9. The composition according to any one of claims 1 to 8, further comprising an active ingredient (c) selected from anticancer agents, analgesics, anesthetics, active molecules having a healing effect, antibacterial agents, antibiotics, bactericidal agents, hemostatic agents, antifungal agents, antiviral agents, antithrombotic agents, anti-inflammatory agents, antipruritic agents, contrast agents, and hormones.

10. The composition according to any one of claims 1 to 9, comprising a compound (d) selected from inorganic nanoparticles, organic-inorganic hybrid nanoparticles, or polymer nanoparticles.

11. A pharmaceutical composition comprising the composition according to any one of claims 1 to 10 and at least one pharmaceutically acceptable excipient.

12. The pharmaceutical composition according to claim 11, wherein the pharmaceutically acceptable excipient is selected from dextran and its derivatives.

13. The pharmaceutical composition according to claim 11, wherein the pharmaceutically acceptable excipient is a hydrophobized dextran that results in the formation of a gel or nanoparticles.

14. A method for preparing the composition according to any one of claims 1 to 10, comprising the step of mixing compounds (a) and (b) in an aqueous solution.

15. A composition according to any one of claims 1 to 10 for use in the treatment of pain.

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