New composition including N-acetylcysteine ​​to alleviate the adverse effects of chemotherapy

Abstract

The invention relates to a composition comprising N-acetylcysteine ​​(NAC) for use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment. It is characterized in that it further comprises Vitamin B12 and / or a combination of Vitamins B1, B2 and B3. The invention relates more particularly to the use of said composition for the prevention and / or treatment of chemotherapy-induced nociceptive and / or neuropathic pain. Figure 9

Description

Title of the invention: New composition comprising N-acetylcysteine ​​to alleviate the adverse effects of chemotherapy Technical field

[0001] The invention relates to a composition comprising N-acetylcysteine ​​(NAC) in combination with other active ingredient(s) for its use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment, in particular the prevention and / or treatment of chemotherapy-induced nociceptive and / or neuropathic pain. Prior art

[0002] N-acetylcysteine ​​or NAC is a non-essential amino acid derived from cysteine, with an acetyl group attached to the nitrogen atom.

[0003] NAC is therefore a thiol that can oxidize by reducing the amount of free radicals. It is also a precursor of glutathione which is a well-known antioxidant.

[0004] It is known to be used as a medicine and as a dietary supplement.

[0005] For example, NAC is commonly used to treat acetaminophen overdoses. NAC may also be useful in other oxidative stress conditions such as apoptosis, mitochondrial dysfunction, neuroinflammation, as well as glutamate and dopamine dysregulation.

[0006] In particular, NAC can be used to improve the symptoms of several types of drug-induced neuropathies.

[0007] Chemo-induced peripheral neuropathies (CIPN) induced for example by platinum salts, taxanes, vinca alkaloids, bortezomib, tha-lidomide and eribulin are a barrier to their use.

[0008] Indeed, these adverse effects, most often dose-dependent and dose-limiting, force clinicians to reduce the dose or even stop treatment prematurely, thus compromising the patient's chances of recovery. The clinical picture of CIPN includes sensory symptoms accompanied or not by motor and / or autonomic symptoms that can be serious and disabling. The modalities of appearance of CIPN are both acute (oxaliplatin and paclitaxel) and chronic.

[0009] Neuropathies induced by anticancer agents may appear immediately but more often they develop gradually and may persist for several years after the end of treatment, and clinical symptoms may become irreversible. This suggests the implementation of a series of mechanisms different, which can range from immediate actions to remodeling and ultimately neuronal damage.

[0010] Table 1 below provides a quick reminder of the different types of neuropathic effects complained of by patients treated with platinum derivatives or taxanes.

[0011] Table 1: Side effects of chemotherapy observed clinically Anticancer Agent Sensory Effects Pain Motor Effects Autonomic Effects Reflexes Recovery Cisplatin Paresthesia, Vibration, Proprioception, Sensitivity, random thermal Dysesthesia Normal Rare (orthostatic dysregulation) Reduced Carbolatin Similar to cisplatin Oxaliplatin (acute reaction) Dysesthesia, cold allodynia, mechanical hyperalgesia Muscle cramps Normal Normal Recovery after a few days Oxaliplatin (chronic) Similar to cisplatin Paclitaxel, Docetaxel Paresthesia, Vibration, Proprioception, Sensitivity, thermal and mechanical Dysesthesia, burning pain, paradoxical sensitivity to heat Rare (proximal > distal weakness) Rare (orthostatic dysregulation) Reduced Generally no recovery and progression possible after stopping treatment

[0012] For short- or medium-term side effects, mechanical, thermal or painful stimuli are in all cases detected and transmitted by ion channels, the main known ones of which are listed in Table 2 below. Cisplatin has been shown to have an acute effect on some of these channels (Milosavljevic et al., Cancer Res., 2011).

[0013] Table 2: Examples of channels involved in nociception and mechanosensation Channels Role Threshold potassium channels TREK1 / TRAAK Neuroprotection, pain, depression TREK2 Polymodal pain / nociception, sensitivity to hot and cold, allodynia to cold associated with oxaliplatin Kvl.1 / 1.2 / 1.4 Peripheral neuropathies Excitatory sodium channels ASICla / ASIC2a / ASIC3 Nociception during tissue acidosis and inflammation Navl.7 / Navl.8 / Navl.9 Allodynia and inflammatory pain Other excitatory channels P2X2-3 / 3 / P2X4 Allodynia and inflammatory pain TRPV1 / TRPM8 / TRPA1 Thermal sensitivity Piezo 2 Fine touch and pain perception Cava2ô-1 (Cavl.2 / 1.3 / Cav2.1-2.3) Sensitivity to touch

[0014] Many preventive and curative treatments have been tested in this indication, namely chemotherapy-induced neuropathies, with varying degrees of success (Thesis by Thibaut Fayolle, “Therapeutic strategy for chemotherapy-induced peripheral neuropathies”, UNIVERSITE CLERMONT AUVERGNE, 2018).

[0015] Examples of compounds used in the prevention and treatment of chemotherapy-induced neuropathies (Emilie Le Rhun, “Clinical characteristics of chemotherapy-induced neuropathies”, CHRU Lille and Unicancer, 2016) include:

[0016] - among the compounds which do not appear to be of preventive interest: lipoic acid, ni- modipine, gabapentin, lamotrigine, acetyl 1-carnitine, venlafaxine;

[0017] - among the compounds showing variable preventive results: amifostine, glutathione, calcium and magnesium infusions, vitamin E;

[0018] - among the compounds presenting a possible preventive interest: N-acetylcysteine, car- bamazepine and oxcarbazepine, Xaliprodenis (IA 5-hydroxytryptamine agonist), Go- shajinkigan, erythropoietin, PFT-u, omega-3 fatty acid, topical gel containing baclofen, amitriptyline, nortriptyline and ketamine, combination of Vitamin B12 / B6;

[0019] - among the pharmaceutical compounds with possible curative interest: du- loxetine;

[0020] - among the pharmaceutical compounds with a questionable curative effect: ga bapentin or pregabalin, venlafaxine, amitriptyline, serotonin and norepinephrine reuptake inhibitors, opioids, local anesthetics, lidocaine infusion, dextromethorphan;

[0021] - among the compounds which do not appear to have a curative effect: lamotrigine, imipramine.

[0022] For this purpose, document WO03045334 is also known, which describes a chemoprotective composition (the chemotherapeutic agent being, for example, cisplatin) comprising at least two chemoprotectants chosen from the group consisting of methionine, N-acetyl-DL-methionine, S-adenosylmethionine, cysteine, homocysteine, cystathione, cysteamine, N-acetylcysteine, glutathione, glutathione ethylester, glutathione, glutathione diethylester triethylester, cysteamine, DiNAC, RibCys, RibCyst, ss-LactCys, a-LactCys, MeliCys, MaltCys, CellCys, OTCA, al-lopurinol, 1-methylallopurinol, 2-methylallopurinol, 5-methylallopurinol 1, 7-methylallopurinol, 7-methylallopurinol, 7-methylallopurinol, 2,5-dimethylallopurinol, 1,7-dimethylallopurinol, 2,7-dimethylallopurinol, 5, 7-dimethylallopurinol, 2,5, 7-trimethylallopurinol, I-ethoxycarbonylallopurinol, I-ethoxycarbonyl-5-methylallopurinol, 2-phenyl-l, 2-benzoisoselenazol-3 (2H)-one and 6-diSeCD.

[0023] Similarly, WO2007044700 relates to a therapeutic combination comprising two or more protective agents selected from a group consisting of specific methionine protective agents, N-acetylcysteine, carnitine, magnesium ions, lipoic acid, ebselen, glutathione and glutathione ester, administered to treat ototoxicity, nephrotoxicity, neurotoxicity, alopecia, gastrointestinal disturbances or reduced survival in a patient undergoing treatment with a chemotherapeutically effective amount of a platinum antitumor compound.

[0024] Furthermore, document US20020058628 discloses a method of treating a disease or condition for which the treatment involves promoting the growth of neuronal cell processes and the therapy of which comprises administering an agonist for neuronal cell surface receptors that promote the growth of neuronal processes, for example chemotherapy-induced neuropathy, said method comprising administering a therapeutically effective amount of said agonist in combination with a therapeutically effective concentration of one or more antioxidants / free radical scavengers and / or an agent capable of increasing intracellular thiol levels and / or a steroid. A representative intracellular thiol is glutathione, the agent capable of raising intracellular thiol levels is N-acetylcysteine ​​(NAC), and representative free radical scavenging antioxidants may be selected from the group consisting of vitamin C, vitamin E, their analogs, and mixtures thereof. A representative vitamin E analog is trolox.

[0025] Finally, it is known that vitamin B12 deficiency can lead to various neurological and behavioral disorders such as ataxia, muscle weakness, incontinence, hypotension, vision disorders, psychotic and mood disorders.

[0026] However, the data appear to be insufficient to conclude that any of the potential chemoprotective agents such as, for example, acetylcysteine, amifostine, calcium and magnesium, diethyldithiocarbamate, glutathione, ORG 2766, oxcarbazepine, retinoic acid or vitamin E, can effectively prevent or limit the neurotoxicity of platinum-based drugs in human patients, as determined using objective quantitative measures of neuropathy (Albers JW et al., “Interventions for the prevention of nerve damage caused by cisplatin and other platinum-based anti-tumor drugs,” Cochrane Database of Systematic Reviews 2014, Issue 3. Art. No.: CD005228. DOI: 10.1002 / 14651858.CD005228.pub4). Technical problem

[0027] Considering the above, a problem which the present invention proposes to solve consists in developing a new combination comprising N-acetylcysteine ​​(NAC) and having a potentiated effect in the prevention and / or treatment of at least one undesirable effect of chemotherapy during cancer treatment. Solution provided

[0028] The solution to this problem posed has as its first object a composition comprising, in a physiologically acceptable medium, N-acetylcysteine ​​(NAC), characterized in that it further comprises Vitamin B12 and / or a combination of Vitamins B1, B2 and B3, for its use in the prevention and / or treatment of at least one undesirable effect of chemotherapy during cancer treatment.

[0029] Surprisingly, among the number of possible combinations of chemoprotective agents, and considering the combinations already envisaged and tested and in particular with N-acetylcysteine ​​(NAC), it is not obvious to obtain a potentiated effect with the specific use of N-acetylcysteine ​​(NAC) combined with Vitamin B12 and / or a combination of Vitamins B1, B2 and B3 to compensate for the effects side effects of chemotherapy. Benefits provided

[0030] The Applicant has notably been able to develop, surprisingly, a composition comprising NAC in combination with other active ingredient(s) which is particularly effective in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment, in particular in the prevention and / or treatment of chemotherapy-induced nociceptive and / or neuropathic pain.

[0031] The composition according to the invention makes it possible more particularly to reduce the useful doses of NAC while maintaining the same effect. Brief description of the drawings

[0032] The invention and the advantages resulting therefrom will be better understood upon reading the description and the non-limiting embodiments which follow, illustrated with reference to the appended drawings in which: Fig.l

[0033] [Fig.l] illustrates the effect of taxanes (A and C) and cisplatin (B and D) on the expression of the Cava2ô-1 subunit (A and B) and the Piezo 2 channel (C and D), involved respectively in the transmission of pain and the detection of mechanical stimuli. Fig.2

[0034] [Fig.2] illustrates the transcription factors targeting genes whose expression is modified after treatment with taxanes and / or cisplatin platinum in primary DRG cultures. Fig.3

[0035] [Fig.3] illustrates the unique kinetics of the inhibition of the genes Piezo 2, CaVa2ô-l and ARNT2 after 0, 4, 8 and 16h of cisplatin treatment (1.25qg / ml) in mouse DRGs from primary culture. Fig.4

[0036] [Fig.4] illustrates the expression of Pizeo 2, Cav a2dl and ARNT2 in neurons in primary culture treated with an oxidizing agent (Tert-Butyl hydroperoxide, TBHP). Fig.5

[0037] [Fig.5] illustrates the dose response of NAC on gene expression of Arnt2, Piezo 2 and CaVa2ô-l after 24h of exposure to 1.25qg / ml of cisplatin. Fig.6

[0038] [Fig.6] illustrates the dose response of Beta Carotene on the gene expression of Arnt2, Piezo 2 and CaVa2ô-l after 24h of exposure to 1.25qg / ml of cisplatin. Fig.7

[0039] [Fig.7] illustrates the dose response of Vitamin B12 on the gene expression of Arnt2, Piezo 2 and CaVa2ô-l after 24h of exposure to 1.25qg / ml of cisplatin. Fig.8

[0040] [Fig.8] illustrates the dose response of Luteolin on the gene expression of Arnt2, Piezo 2 and CaVa2ô-l after 24h of exposure to 1.25qg / ml of cisplatin. Fig.9

[0041] [Fig.9] illustrates the effect of combinations of NAC and different Vitamins on gene expression of Arnt2, Piezo 2 and CaVa2ô-l after 24h exposure to 1.25qg / ml cisplatin. Fig. 10

[0042] [Fig. 10] illustrates the effect of combinations of NAC and Beta Carotene on the expression gene expression of Amt2, Piezo 2 and CaVa2ô-l after 24 hours of exposure to 1.25qg / ml of cisplatin. Description of the embodiments

[0043] In this description, unless otherwise specified, it is understood that, when an interval is given, it includes the upper and lower bounds of said interval.

[0044] The invention relates to a composition comprising, in a physiologically acceptable medium, N-acetylcysteine ​​(NAC) for use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment.

[0045] NAC is a non-essential amino acid derived from cysteine, with an acetyl group attached to the nitrogen atom.

[0046] NAC is therefore a prodrug of the endogenous amino acid L-cysteine, a precursor of glutathione and is known to possess mucolytic, antioxidant and potentially cytoprotective, anticancer and anti-inflammatory activities.

[0047] Upon administration, acetylcysteine ​​exerts its mucolytic activity by reducing disulfide bonds in mucoproteins, resulting in liquefaction of the mucus and reducing its viscosity.

[0048] Its antioxidant activity is attributed to the ability of GSH to scavenge reactive oxygen species (ROS), thereby preventing ROS-induced cell damage, decreasing oxidative stress, protecting cells from the damaging effects of free radicals and preventing apoptosis in these cells.

[0049] Furthermore, it can inhibit tumor cell proliferation, progression and survival, in sensitive tumor cells that depend on ROS-mediated signaling for their proliferation and malignant behavior.

[0050] Under certain circumstances, acetylcysteine ​​is capable of inducing apoptosis in susceptible cells, including certain tumor cells, via the intrinsic mitochondria-dependent pathway but not involving endoreticulum stress. plasma.

[0051] Acetylcysteine ​​also possesses anti-inflammatory activity via modulation of the nuclear factor-kappa B (NF-kB) pathway and modulation of cytokine synthesis.

[0052] The composition according to the invention is characterized in that it further comprises Vitamin B12 and / or a combination of Vitamins Bl, B2 and B3, preferably Vitamin B12 taken alone or in association with the combination of Vitamins Bl, B2 and B3.

[0053] Vitamin B12, also known as cobalamin, is a highly complex, essential water-soluble vitamin that contains the mineral cobalt. This vitamin is produced naturally by intestinal microorganisms and is also found in soil and water. It is necessary for DNA synthesis and cellular energy production.

[0054] Vitamin B12 has many forms, including cyano, methyl, deoxyadenosyl and hydroxy-cobalamin. The cyano form is the most widely used form.

[0055] Advantageously, the composition according to the invention further comprises a combination of vitamins B1, B2 and B3, in different or equal proportions, preferably in equal proportions, taken alone or preferably in association with Vitamin B12.

[0056] Vitamin B1, also called thiamine, is a metabolic precursor of thiamine pyrophosphate (TPP), a coenzyme essential for certain decarboxylases. It is essential for the transformation of carbohydrates into energy by the Krebs cycle and is necessary for the proper functioning of the nervous system and muscles. It is in fact essential for the transformation of pyruvate produced by glycolysis and toxic to the nervous system. It can also inhibit the action of glucose and insulin in the proliferation of arterial smooth muscle cells. Thiamine can also protect against lead toxicity by inhibiting lead-induced lipid peroxidation.

[0057] Vitamin B2 or riboflavin, is a vitamin required for the synthesis of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN), two essential cofactors for flavoproteins. These cofactors are of vital importance in normal tissue respiration, pyridoxine activation, tryptophan to niacin conversion, lipid, carbohydrate, and protein metabolism, and glutathione reductase-mediated detoxification.

[0058] Riboflavin may also be involved in maintaining the integrity of erythrocytes. In addition, it is essential for healthy skin, nails, and hair.

[0059] Niacin, also known as nicotinic acid and vitamin B3, is a water-soluble essential B vitamin that, when administered in high doses, is effective in lowering low-density lipoprotein (LDL) cholesterol levels and increasing high-density lipoprotein (HDL) cholesterol levels, giving this agent unique value in the treatment of dyslipidemia.

[0060] The composition according to the invention is used in the prevention and / or treatment of at least one undesirable effect of chemotherapy during cancer treatment.

[0061] Cancer is a disease characterized by the uncontrolled proliferation of cells, linked to the escape from the regulatory mechanisms which ensure the harmonious development of the organism.

[0062] Cancer starts with a normal cell that transforms and becomes abnormal. This transformation can be due to: constant activation of proto-oncogenes, inactivation of tumor suppressor genes or even inactivation of DNA repair genes.

[0063] The cancers more particularly targeted according to the invention result from solid tumors and are preferably chosen from cancers of the testicle, prostate, ovary, cervix, breast, bladder, ENT tumors, head and neck, esophagus, liver, lung, brain tumors, or even neuroblastomas.

[0064] Today there are several anticancer therapies: surgical treatment, radiotherapy and chemotherapy.

[0065] Chemotherapy is a pharmacological treatment that will target signaling pathways or the tumor microenvironment. It is the most widely used treatment for metastatic cancers. It can be combined with other therapies. Chemotherapy will target cell division and therefore the cell cycle: some molecules in the treatment will be DNA intercalators, others will act on enzymes that control DNA topology, which will disrupt DNA replication, some molecules will target microtubules, and finally other molecules will make adducts between the two strands of DNA, which will be detected as lesions of this molecule and thus lead to cell death through different signaling pathways.

[0066] However, chemotherapy has certain limitations. Indeed, it will also target normal cells and therefore present toxicity towards these cells.

[0067] A physiologically acceptable medium means a medium compatible and suitable for use in contact with human and animal cells, in particular with the skin, mucous membranes and / or appendages, without toxicity, irritation, undue allergic response and the like, and proportionate to a reasonable benefit / risk ratio.

[0068] A physiologically acceptable medium according to the invention may comprise any excipient known and used in the pharmaceutical field and food supplements, compatible with NAC and Vitamins B12 and / or B1, B2 and B3 used according to the invention.

[0069] By way of non-limiting example, mention may be made of solvents, buffers, flavorings, binders, chelating agents, surfactants, thickeners, humectants, moisturizers, preservatives, antioxidants, soothing agents, colorants, perfumes and the like, or a mixture thereof.

[0070] Of course, those skilled in the art will take care to choose the possible compound(s) to be added to these compositions in such a way that the advantageous properties intrinsically attached to the present invention are not or are not substantially altered by the envisaged addition. Their concentration is also chosen so that it does not harm the advantageous properties of the compounds used according to the invention.

[0071] Examples of excipients that may be mentioned include xylitol, shellac, lactose, sorbitol, sucrose, wheat starch, magnesium stearate, lipoic acid, or oils rich in omega 3 such as linseed oil, perilla oil, hemp oil, camelina oil, Inca inchi oil, walnut oil, rapeseed oil, chia oil, Chilean rosehip oil, sea buckthorn oil.

[0072] The composition according to the invention is preferentially used for the prevention and / or treatment of chemotherapy-induced nociceptive and / or neuropathic pain, more preferentially when the chemotherapy is induced by platinum derivatives chosen from cisplatin, oxiplatin and carboplatin, or by taxanes chosen from paclitaxel and docetaxel.

[0073] Taxanes and platinum derivatives are used, as first-line therapy or in combination, in more than 80% of cancers treated with chemotherapy. They are used to treat ovarian, breast, liver, lung, head and neck, testicular, prostate and esophageal cancers.

[0074] Taxanes are a class of diterpenes. The primary mechanism of action of the taxane class is inhibition of microtubule function. Microtubules are essential for cell division, and taxanes block normal cell division. Taxanes thus act as a mitotic spindle poison. They are also believed to be radiosensitizers.

[0075] This class includes paclitaxel, known under the trade name Taxol. Paclitaxel is a molecule produced by endophytic fungi, which is also found in extracts of certain species of yew, to which it gives their high toxicity.

[0076] Paclitaxel will inhibit the depolymerization of microtubules, thus blocking the mitosis mechanism. It is a drug molecule used in the treatment of cancers and injected by perfusion. In oncology, paclitaxel is mainly used in lung cancer, breast cancer and ovarian cancer.

[0077] The side effects are those encountered with anticancer treatments: a Decrease in white blood cells, platelets, and red blood cells, hair loss, and inflammation of the mucous membranes, especially in the mouth. Specific side effects of paclitaxel include peripheral nerve damage, sometimes severe, a risk of fluid retention (ascites, pleural or pericardial effusions), skin reactions, nail changes, and hypersensitivity reactions to the injection.

[0078] Docetaxel is an alkaloid obtained by semisynthesis from a molecule extracted from the European yew tree. Docetaxel is an analogue of paclitaxel, with a similar structure but with double the activity. It differs mainly in its toxicity and its antitumor efficacy. Docetaxel also stabilizes microtubules by inhibiting their depolymerization through stable binding to tubulin and leads to a blockage of mitosis. It is injected intravenously.

[0079] The use of docetaxel leads to bone marrow toxicity, the appearance of edema and effusions due to water retention and neurological disorders. That said, it is effective for quite a number of tumors.

[0080] Platinum salts are a class of platinum-based chemotherapy drugs. This class includes three commonly used molecules: cisplatin, oxaliplatin, and carboplatin. They are often referred to as alkylating agents but essentially make guanine-guanine adducts in the DNA double helix to form the DNA-platinum complex, which will block DNA replication, transcription, and repair. They also have immunosuppressive properties.

[0081] Platinum salts are powerful cytotoxic products. Thus, they cause: - nephrotoxicity which is acute tubular necrosis which can lead to chronic renal failure; - ototoxicity, with possible permanent loss of hearing in high frequencies; - vision problems; - taste disorders; - neurotoxicity: peripheral neuropathies; - neutropenia; - myelosuppression; - frequent nausea and vomiting; and - allergic reactions.

[0082] According to the invention, the term "treatment" means an improvement, prophylaxis or reversal of a disease or disorder, or at least one discernible symptom thereof. It also means an improvement, prophylaxis or reversal of at least one measurable physical parameter related to the disease or disorder being treated, which is not necessarily perceptible to the subject. In another embodiment, the term "treatment" refers to inhibiting or slowing the progression of a disease or disorder, either physically, for example, stabilizing a discernible symptom, physiologically, for example, stabilizing a physical parameter, or both. The term "treatment" also refers to delaying the onset of a disease or disorder. In certain particular embodiments of the invention, the composition of interest is administered as a preventative measure. In this context, the term "prevention" refers to reducing the risk of acquiring a specified disease or disorder.

[0083] More particularly, the applicant was able to demonstrate that the composition used according to the invention is particularly effective for the prevention and / or treatment of chemo-induced nociceptive and / or neuropathic pain.

[0084] Nociceptive pain results from the activation of the nociception system by an internal or external lesion of the body. In oncology, this explains more than 70% of pain syndromes. The lesion can be caused by cancer as well as by cancer treatments.

[0085] Neuropathic pain is the consequence of an injury to the nervous system: compression of a nerve by the tumor, neurological toxicity of chemotherapy, after-effects of surgery for example. They are felt in an unusual way: burning, tingling, electric shocks. When the cause of the neurological suffering disappears, the pain can persist in a transient or chronic manner (painful after-effect). Their diagnosis is sometimes difficult, because these pains can occur with a time lag after the nerve injury. In oncology, neuropathic pain is found, alone or in association with nociceptive pain.

[0086] The different types of neuropathic effects complained of by patients treated with platinum derivatives or taxanes are described in Table 1 above.

[0087] The channels involved in nociception and mechanosensation are described in Table 2 above. The applicant was able to show that the oxidative stress induced by these anticancer agents strongly disrupts the expression of some of these channels, in sensory neuron cultures. He also showed that the addition of a powerful antioxidant that is perfectly well tolerated in humans, NAC, strongly limits this disruption in expression and suppresses the resulting neuropathies in a mouse model.

[0088] As illustrated in example 5, the applicant was able to demonstrate that there is a synergy between NAC and vitamin B12, since the latter makes it possible to reduce the useful doses of NAC while maintaining the same effect.

[0089] The composition used according to the invention comprises NAC at a concentration of between 10 and 90%, for example 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, preferably between 20 and 80%, more preferably between 25 and 75%, for example 25% and 75%, by weight of the total weight of the composition.

[0090] According to a preferred embodiment, the composition used according to the invention comprises vitamin B12 at a concentration of between 0.04 and 0.08%, for example 0.05%, 0.06%, 0.07%, preferably between 0.05 and 0.07%, for example 0.06%, by weight of the total weight of the composition, taken alone or in association with a combination of Vitamins B1, B2 and B3.

[0091] According to another embodiment, the composition used according to the invention comprises a combination of Vitamins B1, B2 and B3, in different or equal proportions, preferably in equal proportions, at a total concentration of between 20 and 90%, for example 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, preferably between 40 and 60%, for example 50%, by weight of the total weight of the composition, taken alone or in association with Vitamin B12, preferably in association with Vitamin B12 at a concentration of between 0.04 and 0.08%, more preferably between 0.05 and 0.07% by weight of the total weight of the composition.

[0092] Advantageously, according to a first embodiment, the composition used according to the invention comprises: - between 50 and 90% of NAC by weight of the total weight of the composition; and - between 0.04 and 0.08% of Vitamin B12 by weight of the total weight of the com position.

[0093] Particularly advantageously, the composition used according to the invention comprises: - between 70 and 80% of NAC by weight of the total weight of the composition; and - between 0.05 and 0.07% of Vitamin B12 by weight of the total weight of the com position.

[0094] Alternatively, according to another embodiment, the composition used according to the invention comprises: - between 10 and 30% of NAC by weight of the total weight of the composition; and - between 20 and 90% of a combination of Vitamins B1, B2 and B3 by weight of the total weight of the composition.

[0095] Advantageously, the composition used according to the invention comprises: - between 15 and 30% of NAC by weight of the total weight of the composition; and - between 30 and 60% of a combination of Vitamins B1, B2 and B3 by weight of the total weight of the composition.

[0096] Alternatively, according to another embodiment, the composition used according to the invention comprises: - between 10 and 90% of NAC by weight of the total weight of the composition; and - between 0.04 and 0.08% of Vitamin B12 and between 20 and 90% of a combination of Vitamins Bl, B2 and B3 by weight of the total weight of the composition.

[0097] Advantageously, the composition used according to the invention comprises: - between 15 and 30% of NAC by weight of the total weight of the composition; and - between 0.05 and 0.07% of Vitamin B12 and between 30 and 60% of a combination of Vitamins Bl, B2 and B3 by weight of the total weight of the composition.

[0098] Preferably, the daily dose of NAC of the composition used according to the present invention is between 200 and 1200 mg and the daily dose of vitamin B12 is between 0.16 and 1 mg and / or the daily dose of the combination of Vitamins B1, B2 and B3 is between 400 and 2400 mg, more preferably between 280 and 1200 mg of NAC, between 0.50 and 1 mg of vitamin B12 and / or between 560 and 2400 mg of the combination of Vitamins B1, B2 and B3.

[0099] This daily dose is preferably administered in 1, 2 or 3 doses (morning, midday and / or evening) for example in the form of 1, 2, 3, 4, 6, 7, 8, 9, 10, 11 or 12 tablets.

[0100] The composition used according to the invention can be administered orally, injectably or transdermally, preferably orally.

[0101] Advantageously, the composition used according to the invention can be administered orally, in one or more identical or different formulations. It is presented in any galenic form normally used for oral administration and in particular in the form of a gelatin capsule, tablet, capsule, soft capsule, dragee, sachet, tube, bottle, chewing gum, ball, emulsion, suspension, liquid, solution, ampoule, drink, syrup, powder for example contained in sachets in particular alu-alu sachets, solid, soft gel, semi-solid.

[0102] Advantageously, the composition used according to the invention is formulated in the form of a tablet, capsule, soft gel, semi-solid, solid, liquid or powder.

[0103] Even more advantageously, the composition used according to the invention is formulated in the form of microgranules with controlled release in capsules.

[0104] Advantageously, the microgranules are coated with a semi-permeable membrane which protects the active substances against degradation and thus controls their release. This membrane will therefore make it possible to mask the taste and / or smell of the substances it coats, the stability of the substances and the protection of the active ingredients against acidity in the stomach. It will also allow control of the release of the active ingredients.

[0105] By way of non-limiting example, the final weight of the capsule is preferably between 200 and 600 mg. Even more preferably, it is between 300 and 500 mg. Particularly advantageously, the final weight of the capsule is 400 mg.

[0106] According to another embodiment, the composition used according to the invention can be administered by injection in liquid form, for example as an adjuvant in a chemotherapy solution.

[0107] According to another embodiment, the composition used according to the invention can be administered transdermally, for example in the form of a patch or medical device capable of increasing penetration through the skin to reach the blood circulation without injection, advantageously with prolonged and continuous release.

[0108] The invention also relates to a product comprising: - as the first active ingredient of NAC, and - as a second active ingredient of vitamin B12 and / or a combination of vitamins Bl, B2 and B3, as a combination product for simultaneous, separate or spread out use over time for its use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment, preferably for the prevention and / or treatment of chemotherapy-induced nociceptive and / or neuropathic pain.

[0109] According to one embodiment, the invention relates to a product comprising: - as the first active ingredient of NAC, - as the second active ingredient of vitamin B12, and as a combination product for simultaneous, separate or spread out use over time for its use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment.

[0110] Alternatively, the invention relates to a product comprising: - as the first active ingredient of NAC, and - as a second active ingredient a combination of vitamins B1, B2 and B3, as a combination product for simultaneous, separate or spread out use over time for its use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment.

[0111] According to another embodiment, the invention relates to a product comprising: - as the first active ingredient of NAC, - as the second active ingredient of vitamin B12, and - as a third active ingredient a combination of vitamins Bl, B2 and B3, as a combination product for simultaneous, separate or spread out use over time for its use in the prevention and / or treatment of at least one adverse effect of chemotherapy during cancer treatment. Examples

[0112] Example 1: Identification of 2 genes whose expression modification is common to the different agents used

[0113] The inventors sought to evaluate the way in which neuropathic side effects begin to set in in the medium term (effects on which it is most interesting to act).

[0114] The inventors have worked more particularly on the variations in expression of the level of expression of the channels which are most likely to explain what happens in this time window.

[0115] From a mechanistic point of view, it is clear that remodeling the expression of different channels depending on the agent used in chemotherapy can give similar symptoms.

[0116] The objective is to identify genes whose remodeling is similar in platinum or taxane chemotherapies.

[0117] Material and Method

[0118] 1. Primary culture of mouse dorsal root ganglia (DRG)

[0119] Model: C57B16 females 8-15 weeks old (Young adult mice that have reached sexual maturity)

[0120] It was chosen to work on female mice because the appearance of chemotherapy-induced peripheral neuropathies is not linked to the sex of the mouse or the patient.

[0121] 24 hours before dissection: - Preparation of solutions i. HBSS-GLUCOSE:

[0122] 50 mL of HBSS 10X

[0123] 1.7 mL HEPES 1.5M

[0124] 10 mL of 0.5M D-Glucose

[0125] 5 mL of Peni-Streptomycin

[0126] QSP to 100mL H2O miliQ

[0127] Adjust PH to 7.5 with NaOH

[0128] QSP 500mL H2O miliQ, Filter and bottle (store at 4°C).

[0129] ii. Neurobasal Medium with NGF:

[0130] For 25mL of medium: 500pL of B27, 250pL of peni-streptomycin, 250pL of L-Glutamine, 6.25pL of NGF (2000pg / mL) and complete to 25mL with Neurobasal medium.

[0131] iii. Enzymatic digestion:

[0132] In 4 mL of HBSS-GLUCOSE, put 80 mg of Collagenase II and 200 mg of Dispase, then aliquot into 2 mL tubes, 200 pL per tube, and freeze. - Coating 12-well plate 24 hours before dissection, a 12-well plate must be coated.

[0133] For 2 hours at room temperature, add 500uL of PLL (Poly-L-Lysine) at 100pg / mL per well; then add 500pL of Laminin at 20pg / mL for 1 hour at 37°C.

[0134] Rinse with HBSS IX and let dry under cover overnight. - Mouse dissection Dissection and extraction of DRGs should not exceed 1 hour to limit cellular damage. The mouse is dissected ventrally and the spine is extracted and then placed under a binocular microscope to extract the DRGs.

[0135] The extraction takes place "dry" (not in cold PBS), the column and the DRGs are often sprinkled with cold HBSS-glucose so as not to let them dry out. After having recovered between 20-30 DRGs in a 2mL tube placed in ice, they are placed under a binocular magnifying glass in order to cut the filaments using a scalpel (only the clusters of neurons are kept).

[0136] The DRGs are then collected in a 15mL tube placed in ice while going into cell culture. - Enzymatic digestion of DRGs DRGs are rinsed twice with HBSS-Glucose at room temperature.

[0137] A 2mL tube containing the enzymatic mixture of Dispase and Collagenase II is recovered to which 50qL of 0.2M CaCl2 is added.

[0138] 1750pL of HBBS-Glucose is then added.

[0139] Put ImL of enzymatic digestion on the DRG and put at 37°C for 20 minutes.

[0140] Put another 1 ml of enzymatic digestion on the DRG and put at 37°C for 20 minutes.

[0141] During the enzymatic digestion time, coat 3 syringes: 21G, 22G and 26G with S VF (Fetal Calf Serum).

[0142] Once the enzymatic digestion is complete, rinse twice with HBSS-Glucose, rinse once with 500qL of Neurobasal medium with NGF.

[0143] Add the required amount of Neurobasal medium with NGF to the wells (200qL / well).

[0144] Pass the DRGs through the 21G to 26G syringes, 6 passes each time.

[0145] Then put 200qL per well and put at 37°C for 1 hour.

[0146] Supplement with 600qL of Neurobasal medium with NGF. Place at 37°C.

[0147] 2. Treatment of DRGs in culture

[0148] The next morning, deposit 1.25 μg / ml of cisplatin with the other molecules to be tested.

[0149] DRG treatment for 24 hours.

[0150] RNA extraction and RT-qPCR

[0151] 3. QPCR - RNA extraction The RNAs are extracted with the “NucleoSpin RNA” kit from Macherey-Nagel.

[0152] The RNAs are then measured using the Nanodrop and stored at -80°C. - Reverse transcription Reverse transcriptions were performed following the protocol of the “SuperScript III Reverse Transcriptase” kit from Thermo Fisher Scientific, doubling the quantities of RNA and products used to obtain a higher concentration of cDNA. qPCR qPCRs were performed with T cDNA obtained by reverse transcription without dilution using 4.625 pL of cDNA and 5.375 pL of SyberROX PCR Mix (final 100 pL per well).

[0153] Each well was made in duplicate. - Analysis of results It is considered that when the cycle threshold (CT or Cycle Threshold) > 35, the gene is not expressed in the analyzed cells.

[0154] To compare our results with or without treatment, we calculated the delta between the target gene and the reference gene (36B4) for each condition (e.g. dCTP1EZ02 = CT piezo2 - CT36B4 = 27.4-21.6 = 5.8 (control); 30.6-23.4 = 7.2 (cisplatin)).

[0155] Then, we did the delta delta CT of the target gene for the test condition and the control condition (ex. ddCT ^02 = dCT Piezo2 (cisplatin) - dCT TREKi(cOntrôie) = 7.2-5.8 = 1.4).

[0156] Finally, we determined the relative expression by calculating: 2A ddCT (e.g. for PIEZO2: 2A ddCT = 2A14 = 0.40).

[0157] In our example, the relative expression of PIEZO2 in the cells treated with cisplatin is therefore 0.40 compared to the control cells whose expression is normalized to 1. - Microscopy In the experiments carried out and exemplified below, a phase contrast microscopy photo (not illustrated) is systematically taken to visualize the effect of the chemotherapy treatment and the possible neuroprotective effect of the molecules tested.

[0158] In this example on gene remodeling induced by cisplatin and taxanes at the level of ion channels expressed in mouse DRG from primary culture, according to the results obtained and illustrated in [Fig.l], 2 genes, namely Piezo 2 and CaVa2ô-l, for which the modification of expression is common to the different agents used, were more particularly identified.

[0159] The 2 genes therefore appear to present a common response to treatment with platinum derivatives and taxanes.

[0160] The expression of Piezo 2 has therefore notably been used as an in- reporter interesting for testing chemoprotective molecules.

[0161] Example 2: Identification of upstream regulators

[0162] By analyzing the promoters of genes whose expression changes are similar, we were able to identify a certain number of transcription factors that could be common regulators, as illustrated in [Fig. 2]. Of course, they must then be tested in parallel using QPCR.

[0163] As illustrated by [Fig.3] relating to a treatment with cisplatin (1.25qg / ml) in mouse DRGs from primary culture, the transcription factor which has a pattern similar to Piezo and CaVa2ô-l is ARNT2.

[0164] This is interesting because it is at the protein level both activated in response to oxidative stress and xenobiotics and will in turn activate genes responding to these stresses.

[0165] This led to the hypothesis that the oxidative stress induced by the chemotherapies tested has a deleterious effect in the medium term on the expression of ARNT2 which amplifies their toxic effect in neurons.

[0166] In contrast, as illustrated by [Fig.3], the effects observed for Mafb are not significant. The expression of mRNAs was measured by qPCR and normalized with the reference gene 36B4. The results correspond to the mean ± SEM.

[0167] Example 3: Test of the effect of oxidative stress

[0168] The expression of Pizeo 2, CaVa2ô-l and ARNT2 was measured in primary cultured neurons treated with an oxidizing agent (Tert-Butyl hydroperoxide, TBHP).

[0169] The expression of Kvl.4 (negative control) is not impacted.

[0170] Mouse DRGs from primary culture were treated with TBHP at 10 or 30 qM or 1.25 qg / ml cisplatin (positive control) versus control (CTRL). mRNA expression was measured by qPCR and normalized to the reference gene 36B4. Results correspond to the mean ± SEM.

[0171] This example is important because it clearly shows, as illustrated by [Fig.4], that there is a similar effect between chemotherapies and an agent producing peroxidation.

[0172] This led to testing antioxidant agents on the expression of identified reporter genes, the most obvious being NAC.

[0173] Example 4: Testing the effect of different molecules taken alone with cisplatin

[0174] The effect of different molecules was tested on the expression of the same reporter genes Pizeo 2, CaVa2ô-l and ARNT2 in QPCR on DRG neurons in primary culture: NAC - Beta Carotene (provitamin A) - Vitamin B12 - Luteolin (Flavone)

[0175] According to the results obtained, NAC appears to have a protective effect on mortality. phology of DRG neurons, and allow to raise ex vivo the expression levels of Piezo2, CaVa2ô-l and ARNT2 in neurons in primary culture (as illustrated in [Fig.5]) but also in vivo on the animal exposed to a chemotherapy protocol (not illustrated). Finally, the administration of NAC appears to dramatically reduce tactile hypersensitivity (in response to mechanical stimuli induced by the Von Frey test) triggered by cisplatin and paclitaxel (not illustrated).

[0176] According to the results obtained and as illustrated by [Fig.6], Beta Carotene appears to have an effect only at a 5qM dose, and not at the lower doses tested (0.3, 1 and 3 qM). Although such a 5 qM dose is a dose 1000 times lower than that useful for NAC, such a dose turns out to be somewhat toxic to neurons in culture (not shown).

[0177] Also, according to the results obtained and as illustrated by [Fig.7], Vitamin B12 does not appear to have a conclusive chemoprotective effect. Vitamin B12 even appears to have a little toxicity at a dose of 50 qM on neuronal morphology (not shown).

[0178] Finally, at the doses used, as illustrated by [Fig.8], Luteolin does not appear to provide any improvement either in gene expression or in neuronal morphology at the cell or network level (not shown).

[0179] Example 5: Testing of combinations of molecules with cisplatin

[0180] According to the results obtained and as illustrated by [Fig.9], in a particularly surprising manner, we note a synergistic effect of Vitamin B12 (at a dose of 20qM) taken in combination with NAC which means that at a dose of 5mM of NAC + Vitamin B12, the combination is as effective in improving the expression of genes, in particular Piezo and CaVa2ô-l, as a dose of NAC of 10 mM taken alone, whereas Vitamin B12 had no effect taken alone at a dose of 25 and 50qM (see [Fig.7]), and has a greater effect than the same dose of NAC (5mM) taken alone.

[0181] However, it is noted that there appears to be a toxic effect of the NAC + Vitamin B12 combination (at a dose of 20qM) when the concentration of NAC (10mM) is doubled.

[0182] Also, we note a very good improvement in morphology and neural networks with the combination NAC (5 mM) + Vitamin B12 (20qM) (not shown).

[0183] Similarly, and surprisingly, we also note a synergistic effect of the combination NAC (5mM) + mixture of Vitamins Bl, B2 and B3 with regard to the improvement of gene expression, in particular Piezo, whereas the mixture of Vitamins B1-B3 has no effect taken alone.

[0184] On the other hand, for comparison purposes, according to the results obtained and as illustrated by the [Fig.10], we note that a combination of NAC (10 mM) and Beta Carotene (5 and 2qM) is rather negative compared to the results obtained with the same molecules taken alone (see respectively Figures 5 and 6).

[0185] In addition, toxicity on neuronal cell morphology can be observed with the combination NAC + Beta Carotene of 5qM (not shown).

[0186] Example 6: Capsule A according to the invention

[0187] According to a preferred embodiment of the invention, the composition is in the form of a capsule, for example of approximately 400 mg. Composition % by weight of the total weight of the composition NAC 75 Vitamin B12 0.06 Excipients (xylitol, shellac) 24.94

[0188] Example 7: Capsule B according to the invention

[0189] According to another preferred embodiment of the invention, the composition is presented in capsule form, for example of approximately 400 mg Composition % by weight of the total weight of the composition NAC 25.02 Vitamin Bl, B2 and B3 (in equal proportions) 50.04 Excipients (xylitol, shellac) 24.94

Claims

Claims

1. Composition comprising, in a physiologically acceptable medium, N-acetylcysteine ​​(NAC), characterized in that it further comprises Vitamin B12 at a concentration of between 0.04 and 0.08% by weight of the total weight of the composition and / or a combination of Vitamins B1, B2 and B3, for its use in the prevention and / or treatment of chemo-induced nociceptive and / or neuropathic pain.

2. Composition for its use according to claim 1, characterized in that the chemotherapy is induced by platinum derivatives chosen from cisplatin, oxiplatin and carboplatin, or taxanes chosen from paclitaxel and docetaxel.

3. Composition for its use according to one of the preceding claims, characterized in that it comprises NAC at a concentration of between 10 and 90% by weight of the total weight of the composition.

4. Composition for its use according to claim 3, characterized in that it comprises NAC at a concentration of between 25 and 75% by weight of the total weight of the composition.

5. Composition for its use according to one of the preceding claims, characterized in that it comprises Vitamin B12 at a concentration of between 0.05 and 0.07% by weight of the total weight of the composition.

6. Composition for its use according to one of the preceding claims, characterized in that it comprises a combination of Vitamins Bl, B2 and B3, in different or equal proportions, at a total concentration of between 20 and 90% by weight of the total weight of the composition.

7. Composition for its use according to claim 6, characterized in that it comprises a combination of Vitamins Bl, B2 and B3, in different or equal proportions, at a total concentration of between 40 and 60% by weight of the total weight of the composition.

8. Composition for its use according to one of the preceding claims, characterized in that it comprises Vitamin B12 taken alone or in association with the combination of Vitamins Bl, B2 and B3.

9. Composition for its use according to one of the preceding claims, characterized in that it is in a form suitable for a

10. oral administration. Composition for its use according to claim 7, characterized in that it is in the form of microgranules with controlled release in capsules.

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