Inhibitors of oxphos and / or of mitochondrial activity for the prevention and / or treatment of neurofibroma

Abstract

The present invention concerns at least one modulator of the metabolism and / or mitochondrial activity, in particular at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway, for use for the prevention and / or treatment of neurofibroma. Combination of such modulators, in particular of an inhibitor of the OXPHOS mitochondrial electron transport chain with an inhibitor of the MVA pathway, as well as pharmaceutical compositions comprising them are also contemplated.

Description

[0001] INHIBITORS OF OXPHOS AND / OR OF MITOCHONDRIAL ACTIVITY FOR THE PREVENTION AND / OR TREATMENT OF NEUROFIBROMA

[0002] The present invention concerns at least one modulator of the metabolism and / or mitochondrial activity, in particular at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway, for use for the prevention and / or treatment of neurofibroma. Combination of such modulators, in particular of an inhibitor of the OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway, as well as pharmaceutical compositions comprising them are also contemplated.

[0003] Neurofibromatosis type 1 (NF1) is a complex autosomal dominant disorder caused by germline mutations in the NF1 tumour suppressor gene that encodes the protein neurofibromin. The manifestations of NF1 are widespread and can involve multiple organ systems, such that NF1 patients have an increased risk for skeletal abnormalities, brain and peripheral nerve tumors, learning disabilities, attention deficits, and social and behavioral problems. NF1 is primarily characterized by pigmentary lesions (cafe-au-lait macules, skinfold freckling and Lisch nodules) and neurofibromas - benign peripheral nerve tumors that can either be cutaneous (dermal) or more profound (plexiform and spinal). Dermal neurofibromas are generally superficial, while plexiform neurofibromas are associated with multiple nerve bundles and are located deeper within tissues. Although dermal neurofibromas are not malignant, these benign tumors can cause itching, pain, hemorrhage and a cosmetic burden that has been linked with psychosocial challenges.

[0004] In addition to being painful, plexiform neurofibromas and spinal neurofibromas can cause neurological deficits (including sensory and motor loss) and even vertebral erosion. While dermal neurofibromas do not become malignant, plexiform neurofibromas, which often cause substantial morbidity, can transform in sarcomas known as malignant peripheral nerve sheath tumors (MPNSTs). Overall, neurofibromas are key contributors to reduced life expectancy in NF1 patients and greatly affect quality of life of patents and their families.

[0005] NF1 is a rare disease, with an average global prevalence of -1 / 3,000 (Gutmann D., et al. Nature Reviews Disease Primers, 2017). Notably, the life expectancy of NF1 patients is on average reduced by 8-21 years and the occurrence of death before 40-years old is significantly higher than in the general population.

[0006] At present, no definitive treatment for neurofibromas is available, and clinical management involves surgical removal, laser ablation for small lesions, electrodessication, emollients and make-up, pain management and psychological support. Routine care includes patient-centered longitudinal surveillance with age-specific monitoring of clinical manifestations, aiming at the early recognition and symptomatic treatment of complications as they occur. Active engagement and an active partnership among multiple health care providers, concerned lay groups and patient experts is the cornerstone of management of this rare disease and this complex, and often costly, management program represents a heavy burden for the patient and for the healthcare system.

[0007] Ongoing trials are examining the efficacy of targeted therapies, namely drugs targeted to the specific cellular signaling pathways that are disrupted by the genetic mutation that give rise to NF1 , notably the RAS / MAPK pathway which is negatively regulated by neurofibromin and is thus hyperactivated in NF1 patients. One such therapy, selumetinib (marketed by AstraZeneca under the name Koselugo), has recently been approved by the FDA for the treatment of pediatric NF1 patients with symptomatic, inoperable plexiform neurofibromas. Phase I and phase II trials of selumetinib in children presenting with NF1 , leads to a partial reduction (8-39%) of plexiform neurofibroma tumor volume in a majority of patients (68%), but not complete tumor regression even under a continuous dosing cycle (every 12-hours for 28-day cycles during 1-year).

[0008] Thus, while the results of selumetinib are promising, given that the indication is limited to children, the dosing regime is quite heavy and the lack of full tumor regression indicates the need for the identification and development of other therapeutic options.

[0009] Moreover, due to the fact that neurofibromas can be molecularly heterogeneous, combinations of therapeutic agents could be beneficial and increase efficiency, to target different disease mechanisms, to avoid the development of treatment resistance and to reduce off-target effects (as a consequence of decreased dosing regimens).

[0010] The inventors have now discovered that targeting the metabolism and / or the mitochondrial activity, in particular the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway, can be efficient to prevent and / or treat neurofibroma.

[0011] The present invention thus relates to at least one modulator of the metabolism and / or mitochondrial activity, in particular at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway, for use for the prevention and / or treatment of neurofibroma.

[0012] It further relates to an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway for use for the prevention and / or treatment neurofibroma. It also relates to a pharmaceutical composition comprising an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway.

[0013] Detailed description

[0014] As mentioned, the present invention is directed to at least one modulator of the metabolism and / or mitochondrial activity, in particular at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway, for use for the prevention and / or treatment of neurofibroma.

[0015] By “at least one” is meant one, two, or more, in particular one or two.

[0016] By “one modulator of the metabolism and / or mitochondrial activity” is meant that the action is directed toward the metabolism or the mitochondrial activity, or both.

[0017] The modulator will regulate or adjust to a certain degree the metabolism and / or the mitochondrial activity.

[0018] By “metabolism activity” is meant the set of chemical reactions within a cell that convert metabolic substrates into usable energy (in the form of adenosine triphosphate) for essential cellular processes. .

[0019] By “mitochondrial activity” is meant the biochemical processes and functions carried out by mitochondria within a cell, primarily involving the production of adenosine triphosphate through oxidative phosphorylation.

[0020] In particular, the modulator can be an inhibitor of the metabolism and / or mitochondrial activity.

[0021] For example, it is an inhibitor of one or more of the following genes: AKIRIN 2, GMPS, HSPA8, KANSL1 , MCM3AP, NDUFV1 , WTAP, MVK, RARS2, SLC35A1 and ISCA2, preferably of one or more of the following genes: HSPA8, KANSL1 , NDLIFV1 , MVK, RARS2 and ISCA2.

[0022] By “one or more” is meant that it will either inhibit one the above mentioned genes or several, for example two, three, four, five, six or the totality of them.

[0023] By “inhibitor” is meant that the expression of the gene will be altered or silenced and / or that the product (RNA or protein) of the gene will be altered or silenced.

[0024] The genes mentioned herein are known by the man skilled in the art. In particular:

[0025] - AKIRIN 2 enables enzyme binding activity and identical protein binding activity and is predicted to be involved in positive regulation of innate immune response and positive regulation of transcription by RNA polymerase II;

[0026] GMPS or guanine monophosphate synthase, codes for an enzyme that converts xanthosine monophosphate to guanosine monophosphate;

[0027] HSPA8 : chaperon: plays a critical role in mitochondrial import ; KANSL1 : was shown to be part of the same lysine acetylase complex, it is an important regulator of PINK1-mediated mitochondrial quality control;

[0028] MCM3AP encodes minichromosome maintenance complex component 3 associated protein, one of the MCM proteins essential for the initiation of DNA replication;

[0029] NDLIFV1 : encodes the 51-kD subunit of complex I (NADH:ubiquinone oxidoreductase) of the mitochondrial respiratory chain;

[0030] - WTAP: encodes a re-mRNA-splicing regulator WTAP protein;

[0031] MVK: encodes an enzyme that converts mevalonic acid into mevalonate- 5- phosphate;

[0032] RARS2: encodes a protein that localizes to the mitochondria, where it catalyzes the transfer of L-arginine to its cognate tRNA, an important step in translation of mitochondrially-encoded proteins;

[0033] SLC35A1 : the protein encoded by this gene is found in the membrane of the Golgi apparatus, where it transports nucleotide sugars into the Golgi;

[0034] ISCA2: the protein encoded by this gene is an A-type iron-sulfur cluster (ISC) protein found in mitochondria. The encoded protein appears to be involved in the maturation of mitochondrial iron-sulfur proteins.

[0035] In the context of the invention, the least one modulator described herein is chosen from inhibitors of the OXPHOS mitochondrial electron transport chain and inhibitors of the MVA pathway.

[0036] The OXPHOS mitochondrial electron transport chain is known by the man skilled in the art and refers to a metabolic pathway in which enzymes oxidize nutrients to release stored chemical energy in the form of ATP (adenosine triphosphate). OXPHOS consists of five protein complexes l-V and takes place in the mitochondria.

[0037] The MVA pathway is also well known by the man skilled in the art. The MVA or mevalonate pathway also known as the isoprenoid pathway or 3-hydroxy-3-methylglutaryl- CoA reductase (HMGCR) pathway is an anabolic pathway providing metabolites for multiple cellular processes in eukaryotes, archaea, as well as some bacteria, thus underscoring its importance for nearly all living organisms including humans.

[0038] In particular, the inhibitor of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway are direct inhibitors of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway. By “direct” inhibitor is meant compounds that directly inhibit enzymes involved in these chain / pathway.

[0039] In one embodiment, they are chosen from: (i) inhibitors of complex I of the OXPHOS mitochondrial electron transport chain, and

[0040] (ii) inhibitors of Protein Complex 1 and OXPHOS activity.

[0041] In particular, said inhibitors are chosen from IACS-010759, statins, Zoledronic Acid, and / or the compound IM 156.

[0042] IACS-010759 (IACS), is a potent and selective inhibitor of complex I of the OXPHOS mitochondrial electron transport chain that is currently in phase I trials for the treatment of solid tumors.

[0043] IM 156, is a novel Protein Complex 1 and OXPHOS activity inhibitor that is in preclinical studies for the treatment of solid tumors.

[0044] By “statins” is meant the compounds of the statin family (statins and statin derivatives) which are named with the suffix “statin” and are part of the therapeutic chemistry family of statins. Statins are well known by the person skilled in the art and can be chosen from: pitavastatin, pravastatin, fluvastatin, rosuvastatin, atorvastatin and cerivastatin, pivastatin being particularly contemplated.

[0045] Pitavastatin, is an inhibitor of the MVA pathway that is FDA-approved as an anticholesterol drug.

[0046] In one embodiment, a combination of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway is used to prevent and / or treat neurofibroma.

[0047] More particularly, the combination between IACS-010759, and a statin, more particularly pitavastatin, is contemplated.

[0048] As already mentioned, the present invention also relates to an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway for use for the prevention and / or treatment neurofibroma, in particular IACS-010759, and a statin, more particularly pitavastatin.

[0049] Products comprising an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway as a combined preparation for use spread out over time for the prevention and / or treatment of neurofibroma are also contemplated.

[0050] In particular, the products comprise IACS-010759, and a statin, more particularly pitavastatin.

[0051] The present invention further relates to a pharmaceutical composition comprising an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway, in particular IACS-010759, and a statin, more particularly pitavastatin.

[0052] As mentioned herein the at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway according to the invention, the different combinations described herein and in particular the combination of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway, the products comprising an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway, are used to prevent and / or treat neurofibroma.

[0053] The present invention further relates to a method to prevent and / or treat neurofibroma in a subject in need thereof, said method comprising the administration of an effective amount of at least one modulator of the metabolism and / or mitochondrial activity, in particular at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway, as described herein.

[0054] It also relates to a method to prevent and / or treat neurofibroma in a subject in need thereof, said method comprising the administration of an effective amount of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway as described herein.

[0055] “Neurofibroma”, in the context of the present invention, has the normal meaning for a man skilled in the art. It corresponds to a type of peripheral nerve tumor that forms soft bumps on or under the skin.

[0056] As already mentioned, neurofibromatosis type 1 (NF1) is a complex autosomal dominant disorder caused by germline mutations in the NF1 tumor suppressor gene that encodes the protein neurofibromin. NF1 is primarily characterized by pigmentary lesions (cafe-au-lait macules, skinfold freckling and Lisch nodules) and neurofibromas - benign peripheral nerve tumors that can either be cutaneous (dermal) or more profound (plexiform and spinal).

[0057] Dermal neurofibromas are generally superficial, while plexiform neurofibromas are associated with multiple nerve bundles and are located deeper within tissues. Although dermal neurofibromas are not malignant, these benign tumors can cause itching, pain, hemorrhage and a cosmetic burden that has been linked with psychosocial challenges. In addition to being painful, plexiform neurofibromas and spinal neurofibromas can cause neurological deficits (including sensory and motor loss) and even vertebral erosion. While dermal neurofibromas do not become malignant, plexiform neurofibromas, which often cause substantial morbidity, can transform in peripheral nerve sarcomas known as malignant peripheral nerve sheath tumors (MPNSTs)

[0058] In one embodiment of the invention, the least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway as described herein, the different combinations described herein and in particular the combination of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway, the products comprising an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway, are used for the prevention of the malignant development of neurofibroma.

[0059] In particular, neurofibroma is cutaneous and / or plexiform neurofibroma.

[0060] The expression “effective amount” is intended to mean an amount sufficient to prevent and / or treat a given disease. It will be appreciated that this amount will vary with the effectiveness of therapeutic agent(s) employed, with the nature of any carrier used, with the seriousness of the disease and the age of the patient. The determination of appropriate amounts for any given composition is within the skill in the art, through standard series of tests designed to assess appropriate therapeutic levels.

[0061] The terms "treat", “treating”, “treated” or "treatment", as used in the context of the invention, refer to therapeutic treatment wherein the object is to eliminate or lessen symptoms. Beneficial or desired clinical results include, but are not limited to, elimination of symptoms, alleviation of symptoms, diminishment of extent of condition, stabilized (i.e., not worsening) state of condition, delay or slowing of progression of the condition.

[0062] The terms “prevent”, “prevention”, “preventing” or “prevented”, as used in the context of the present invention, refer to the prevention of the onset, recurrence or spread of a disease or disorder or infection, or of one or more symptoms thereof. In certain embodiments, the terms refer to the treatment with or administration of a compound provided herein prior to the onset of symptoms, particularly to patients at risk of disease or disorder or infection provided herein. The terms encompass the inhibition or reduction of a symptom of the particular disease or disorder or infection. Subjects with familial history of a disease or disorder or infection in particular are candidates for preventive regimens in certain embodiments. In addition, subjects who have a history of recurring symptoms are also potential candidates for the prevention. In this regard, the term “prevention” may be interchangeably used with the term “prophylactic treatment”.

[0063] In particular, the subject in need of a prevention or treatment against neurofibroma is a subject afflicted with such disease.

[0064] In the context of the present invention, the identification of the subjects who are in need of treatment of herein-described diseases and conditions is conducted as above mentioned and is well within the ability and knowledge of the man skilled in the art. A clinician skilled in the art can readily identify, by the above-mentioned technics, those subjects who are in need of such treatment.

[0065] The term “patient" or “subject” refers to a warm-blooded animal such as a mammal, in particular a human, male or female, unless otherwise specified, which is afflicted with, or has the potential to be afflicted with one or more diseases and conditions described herein. In particular, the inhibitors of the combinations according to the invention are administered separately, sequentially or simultaneously.

[0066] The inhibitors provided herein can be formulated into pharmaceutical compositions, optionally by admixture with one or more pharmaceutically acceptable excipients.

[0067] The present invention thus also relates to a pharmaceutical composition comprising at least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway as defined herein, and a pharmaceutically acceptable excipient.

[0068] Such compositions may be prepared for use in oral administration, particularly in the form of tablets or capsules, in particular orodispersible (lyoc) tablets; or parenteral administration, particularly in the form of liquid solutions, suspensions or emulsions.

[0069] It may be prepared by any of the methods well known in the pharmaceutical art, for example, as described in Remington: The Science and Practice of Pharmacy, 20th ed.; Gennaro, A. R., Ed.; Lippincott Williams & Wilkins: Philadelphia, PA, 2000. Pharmaceutically compatible binding agents and / or adjuvant materials can be included as part of the composition. Oral compositions will generally include an inert diluent carrier or an edible carrier. They can be administered in unit dose forms, wherein the term “unit dose” means a single dose which is capable of being administered to a patient, and which can be readily handled and packaged, remaining as a physically and chemically stable unit dose comprising either the active compound itself, or as a pharmaceutically acceptable composition.

[0070] The tablets, pills, powders, capsules, troches and the like can contain one or more of any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, or gum tragacanth; a diluent such as starch or lactose; a disintegrant such as starch and cellulose derivatives; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, or methyl salicylate. Capsules can be in the form of a hard capsule or soft capsule, which are generally made from gelatin blends optionally blended with plasticizers, as well as a starch capsule. In addition, dosage unit forms can contain various other materials that modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents. Other oral dosage forms syrup or elixir may contain sweetening agents, preservatives, dyes, colorings, and flavorings. In addition, the active compounds may be incorporated into fast dissolve, modified-release or sustained-release preparations and formulations, and wherein such sustained-release formulations are preferably bi-modal.

[0071] Liquid preparations for administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. The liquid compositions may also include binders, buffers, preservatives, chelating agents, sweetening, flavoring and coloring agents, and the like. Non-aqueous solvents include alcohols, propylene glycol, polyethylene glycol, acrylate copolymers, vegetable oils such as olive oil, and organic esters such as ethyl oleate. Aqueous carriers include mixtures of alcohols and water, hydrogels, buffered media, and saline. In particular, biocompatible, biodegradable lactide polymer, lactide / glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be useful excipients to control the release of the active compounds. Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer's dextrose, and the like.

[0072] Examples of modes of administration include parenteral e.g. subcutaneous, intramuscular, intravenous, intradermal, as well as oral administration.

[0073] In the context of the present invention, it should be understood that “an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway for use for the prevention and / or treatment of” is equivalent to "the use of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway for the prevention and / or treatment of” and to "the use of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway for the manufacture of a medicament for the prevention and / or treatment of ”.

[0074] The invention will be further illustrated in view of the following figures and examples.

[0075] FIGURES

[0076] Figure 1. Drug tests.

[0077] A: ETC complex I inhibitors (IACS and IM156) and HMG-CoA reductase inhibitor (Pitavastatin) exhibit specific cytotoxicity in NF1-Z- versus WT human Schwann cells. The FDA-approved MEK1 / 2 inhibitor (Selumetinib) is used as a positive control.

[0078] B: The test of IACS, IM156, and Pitavastatin in the same human Schwann cell line with (i) a NF1 heterozygous inactivation (NF1+ / -) or (ii) a NF2 inactivation, confirms the treatment specificity for the complete inactivation of NF1 (NF1

[0079] C: Efficacy of Selumetinib, IACS-10759, and Pitavastatin in primary human Schwann cells (NF1- / -) isolated directly from surgically resected neurofibromas from two NF1 patients.

[0080] EXAMPLES

[0081] MATERIAL AND METHODS

[0082] Drugs tests Immortalized Human Schwann cells (iHS) (hTERT ipnO2.3-ATCC CRL-3392), and primary cell lines were cultivated in adherence in Dulbecco’s modified Eagle’s media (DMEM) supplemented with 10% FBS and 1 % l-glutamine.

[0083] Inactivation of NF1 {NF1+I- or NF1- / -) or NF2 in iHS cells were performed using the genome editing CRISPR / CAS9 technology.

[0084] For the drugs test, 5000 cells (iHS, and primary cell lines) per well were plated into 96-well plates.

[0085] Selumetinib (MedChemExpress, ref. HY-50706), Pitavastatin (Merk, ref. SML2473), and IACS-010759 (Euromedex, ref. TA-T5337) were resuspended in DMSO at the concentration of 10mM and used at the indicated concentrations.

[0086] Plates were measured 48 hr post-treatment using Neutral Red Assay Kit - Cell Viability I Cytotoxicity (Abeam, ab234039).

[0087] PROTOCOL AND RESULTS

[0088] Neurofibromatosis type 1 (NF1) is a genetic disorder caused by heterozygous germline mutations in the NF1 gene, leading to a loss of its normal function. Patients with NF1 initially possess one normal NF1 allele but it is susceptible to inactivation. When this inactivation occurs in Schwann cells or their progenitors, it triggers the formation of benign peripheral nerve sheath tumors called neurofibromas. Neurofibromas can cause a range of health complications, including the potential transformation into life-threatening malignant tumors. The approach focuses on selectively eliminating cells that have lost both copies of the NF1 gene, rather than attempting to correct the disease-causing mutation. This strategy capitalizes on the concept that the mutation of one gene often creates vulnerabilities in other genes that are not typically essential for cell survival.

[0089] To implement this approach, CRISPR-CAS9 gene editing technology was employed to genetically inactivate NF1 in human Schwann cells, the cells from which neurofibromas originate. Isogenic pairs of cells (wild-type versus NF1- / -) were established and extensive genetic screens conducted using genome-wide dropout CRISPR-Cas9 techniques, alongside transcriptomic analysis in these models. Through the research, a previously unrecognized essentiality linked to the control of mitochondrial activity and cellular metabolism by NF1 was unveiled.

[0090] The screening process involved a rigorously tested genome-wide library, ensuring high specificity and efficiency in single-guide RNA selection -critical criteria when exploring gene essentiality. The screens were performed with biological replicates to ensure robust identification of essential genes in human / VF7-mutant versus wild-type Schwann cells. The same screen was performed on a second pair of isogenic epithelial cells (WT vs NF1- / -) to ensure that the vulnerabilities identified in Schwann cells were cell-specific. After primary screens, secondary validations and further analysis on additional relevant cellular models, a highly stringent list of 11 candidate genes that were more essential in the context of NF1- mutated Schwann cells compared to wild-type cells was identified.

[0091] Subsequently, it was investigated whether the unique metabolism of A / Fl-mutant Schwann cells could render them more susceptible to specific mitochondrial and metabolic drugs. The pharmacological tests were conducted in vitro, using relevant neurofibroma models. While selumetinib, an anti-MEK drug, is the only FDA-approved treatment for inoperable pediatric neurofibromas, it only leads to a partial reduction in tumor volume in most patients. Recognizing the need for more effective treatments, various clinical-grade drugs were tested, including statins that inhibit HMG-CoA reductase and recent direct mitochondrial inhibitors such as IACS-010759 and IM156.

[0092] Interestingly, it was found that Pitavastatin, IACS-010759, and IM156 exhibited cytotoxicity specific to A / Fl-mutant Schwann cells, with Pitavastatin and IACS showing even stronger effects than selumetinib (Fig. 1A).

[0093] Considering the potential for treatment resistance and long-term side effects, combination therapy could prove highly beneficial.

[0094] To validate the effectiveness and specificity of the candidate treatment approaches, the response of other Schwann cell models characterized by heterozygous inactivation of NF1 (NF1+ / -) or NF2 genes was analyzed (Fig. 1 B). The candidate drugs were also tested on primary human Schwann cells (NF1- / -) isolated directly from surgically resected neurofibromas from two NF1 patients (Fig. 1C).

[0095] These experiments confirmed the strong specificity of the treatment for NF1-Z- Schwann cells.

[0096] As a further step, the synergy between these drugs and selumetinib, uncovering modest synergy in cytotoxic effects with selumetinib and the three mitochondrial-related drugs were explored, but a patent effective synergy between Pitavastatin and IACS.

Claims

CLAIMS1. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use for the prevention and / or treatment of neurofibroma.

2. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to claim 1 wherein said inhibitor of the OXPHOS mitochondrial electron transport chain is chosen from (i) inhibitors of complex I of the OXPHOS mitochondrial electron transport chain and (ii) inhibitors of Protein Complex 1 and OXPHOS activity.

3. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to claim 1 or 2, wherein said inhibitor is chosen from IACS-010759, statins, Zoledronic Acid, and / or the compound IM 156.

4. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to claim 3, wherein statins are chosen from: pitavastatin, pravastatin, fluvastatin, rosuvastatin, atorvastatin and cerivastatin.

5. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to any one of the preceding claims, wherein a combination of an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway is used to prevent and / or treat neurofibroma.

6. Pharmaceutical composition comprising an inhibitor of OXPHOS mitochondrial electron transport chain and an inhibitor of the MVA pathway.

7. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to claim 5or pharmaceutical composition according to claim 6, wherein the inhibitor of OXPHOS mitochondrial electron transport chain is IACS-010759, and the inhibitor of the MVA pathway is a statin, in particular pitavastatin.

8. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to any of the claims 1-5 and 7, for the prevention of the malignant development of neurofibroma.

9. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to any of the claims 1-5 and 7-8, wherein neurofibroma is cutaneous, and / or plexiform neurofibroma.

10. At least one inhibitor of the OXPHOS mitochondrial electron transport chain and / or inhibitor of the MVA pathway for use according to any of the claims 1-5 and 7-8 or pharmaceutical composition according to claim 6, wherein inhibitors of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway are direct inhibitors of the OXPHOS mitochondrial electron transport chain and / or of the MVA pathway.

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