(19) TZZ¥_ _T (11) EP 3 412 294 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 12.12.2018 Bulletin 2018/50 A61K 31/52 (2006.01) G01N 33/00 (2006.01) (21) Application number: 17174916.1 (22) Date of filing: 08.06.2017 (84) Designated Contracting States: (72) Inventor: JACOB, Claire AL AT BE BG CH CY CZ DE DK EE ES FI FR GB 4051 Basel (CH) GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR (74) Representative: Schneiter, Sorin et al Designated Extension States: Omnis IP SA BA ME Ch. de Champ-Colomb 7 B Designated Validation States: 1024 Ecublens (VD) (CH) MA MD Remarks: (71) Applicant: Universite De Fribourg Claim 16. is deemed to be abandoned due to 1700 Fribourg (CH) non-payment of the claims fee (Rule 45(3) EPC). (54) HDAC1/2 ACTIVATOR FOR PROMOTING AND/OR ACCELERATING MYELINATION AND/OR REMYELINATION (57) The present invention is based on the surprising tems. The present inventor found that HDAC2 finding of the property of promoting myelination of acti- deacetylates eEFlAl and thereby prevents the latter from vators of histone deacetylase (HDAC) 1 and 2. In partic- removing outside the nucleus key inducers of myelin ular, such activators have the capacity of accelerating genes transcription. The HDAC1/2 activators are useful and increasing remyelination after lesions to the myelin in the treatment of diseases associated with demyelina- of nerve cells of the peripheral and central nervous sys- tion, such as Multiple Sclerosis. EP 3 412 294 A1 Printed by Jouve, 75001 PARIS (FR) 1 EP 3 412 294 A1 2 Description Schwann cells convert into repair cells that foster axonal regrowth, and redifferentiate to rebuild myelin. The au- Technical Field thors report that short-term HDAC1/2 inhibitor treatment early after lesion acceleration functional recovery and [0001] The present invention relates to an histone5 enhances regeneration of the crushed nerve cells of the deacetylase 1 and/or 2 (HDAC1/2) activator for promot- PNS. ing myelination, for accelerating remyelination after le- [0007] More specifically, Brügger et al, 2016 (ref. 1) sions to nerve cells, for treating diseases associated with have shown that HDAC2 interacts with the transcription demyelination, to methods for screening agents useful factor Sox10 and recruits histone H3 lysine 9 (H3K9) in the treatment of such diseases, and to methods of 10 demethylases (HDMs) to form a multifunctional protein treatment. complex that de-represses the Sox10 target genes Oct6 and Krox20 to allow their subsequent activation at differ- Background Art and Problems Solved by the Inven- ent time points of the regeneration process after lesion. tion Inactivating this mechanism was shown to result in earlier 15 conversion into repair Schwann cells after lesion and [0002] Multiple Sclerosis (MS) is a demyelinating dis- faster axonal regeneration. ease in which the insulating sheaths of nerve cells in the [0008] It is an objective underlying the present inven- brain, optic nerves and/or spinal cord are damaged. This tion to further elucidate the role of HDAC2 in the regen- damage disrupts the ability of parts of the nervous system eration process after lesion. It is an objective of the to communicate, resulting in a range of signs and symp- 20 present invention to elucidate the mechanisms control- toms, including physical and mental problems. MS is the ling Schwann cell conversion into repair cells and dedif- most frequent degenerative disease of the central nerv- ferentiation after lesion. ous system (CNS), the onset of which is often between [0009] It is a further objective underlying the present 20 and 50 years old. While there are treatments allowing invention to provide novel therapeutic strategies for the alleviating the symptoms of the disease, there is pres- 25 treatment of diseases and neural damages associated ently no cure for MS. There are several factors reported with demyelination in both, the PNS and the CNS. In par- to trigger the onset, such as viral infection, food, genetics ticular, it is an objective of the invention to provide a treat- and environmental factors. ment of MS and other demyelinating diseases. It is also [0003] On the cellular level, MS starts with attacks af- an objective of the invention to provide compounds ca- fecting oligodendrocytes, the cells that produce the my- 30 pable of promoting, accelerating and/or increasing remy- elin sheaths of axons in the CNS. These attacks are elination. It is an objective to provide novel assays for caused, at least in part, by a person’s own immune sys- screening compounds suitable to prevent or treat demy- tem, which is why MS is believed to be an immune-me- elination and for promoting remyelination. diated disorder. Following an attack of the myelin [0010] US 6,277,402 discloses a method for treating sheaths, new oligodendrocytes are generally recruited 35 multiple sclerosis, the method comprising administering to the site of lesion, where they regenerate the myelin a histamine H2 agonist and a phosphodiesterase inhib- sheath in a process referred to as remyelination. Recur- itor. The present invention is not concerned with hista- rent attacks and regeneration result in the pattern of suc- mine H2 agonists and phosphodiesterase inhibitors. cessive relapsing and remitting phases typical for early [0011] US2003/0134865 discloses a method for stages. MS evolves later into a progressive neurodegen- 40 screening comprising the step of exposing a xanthine erative disease where neurons are lost. Indeed, in this compound to a histone deacetylase. According to this later stage, remyelination does no longer occur or is not reference, screened compounds are useful in treating effective, resulting in neuronal loss. conditions caused by or exhibiting abnormal cellular pro- [0004] MS is one of a series of demyelination disor- liferation or differentiation, such as cancer, or inflamma- ders. The latter can affect the CNS as well as the periph- 45 tion, in particular asthma or other inflammatory airway eral nervous system (PNS). For example, demyelination disease, such as COPD (Chronic obstructive pulmonary occurs after traumatic lesions of the CNS and the PNS. disease). The present invention is not concerned with [0005] The myelin sheath, which serves to increase these conditions. nerve conduction velocities, is deposited around axons by specialized cells in the central and peripheral nervous 50 Summary of the Invention systems of higher vertebrates. Indeed, myelin is synthe- sized by oligodendrocytes in the CNS (as already men- [0012] Remarkably, the present inventor provides ev- tioned above) and by Schwann cells in the PNS. idence for the biological activity of activators of HDAC1/2, [0006] Brügger et al, 2016 (ref. 1) have reported that, in particular HDAC2, with respect to the myelination and after sciatic nerve crush lesion, HDAC2 coordinates the 55 remyelination of nerve cells in both, the PNS and the action of other chromatin-remodelling enzymes to induce CNS. the upregulation of Oct6, a key transcription factor for the [0013] In an aspect, the invention provides an activator development of Schwann cells. After injury, mature of HDAC (histone deacetylase) 1 or 2 (increasing 2 3 EP 3 412 294 A1 4 HDAC1/2 activity and/or expression) for increasing, ac- Brief Description of the Drawings celerating and/or promoting myelination. [0014] In an aspect, the invention provides an activator [0022] of HDAC (histone deacetylase) 1 or 2 (increasing HDAC1/2 activity and/or expression) for increasing, ac- 5 Figure 1A shows immunofluorescence of acetylated celerating and/or promoting remyelination. eEF1A (left images), acetylated eEF1A and S100 [0015] In an aspect, the invention provides an activator proteins (center images) and the combined fluores- of HDAC (histone deacetylase) 1 or 2 (increasing cence of DAPI, acetylated eEF1A and S100 proteins HDAC1/2 activity and/or expression) for increasing, ac- (right side images), in sciatic nerve preparations ob- celerating and/or promoting remyelination after lesions 10 tained from four-day old control mice (top row) and to myelin of nerve cells. HDAC1/2 conditional (Schwann cell-specific) knock- [0016] In an aspect, the invention provides an activator out mice (dKO). The bottom left image shows accu- of HDAC (histone deacetylase) 1 or 2 (increasing mulation of acetylated eEF1A in the knock-out mice, HDAC1/2 activity and/or expression) for treating and/or demonstrating that HDAC1/2 deacetylate eEF1A preventing a demyelinating disease. 15 during developmental myelination. [0017] In an aspect, the invention provides an activator of HDAC (histone deacetylase) 1 or 2 (increasing Figure 1B shows an IP (immunoprecipitation) of HDAC1/2 activity and/or expression) for treating and/or eEF1A1 and Western blot (WB) image obtained from preventing one or more condition selected from the group sciatic nerve lysates showing the accumulation of consisting of: (1) traumatic injury of the PNS, (2) multiple 20 acetylated eEF1A1 (Ace-eEF1A1) in the dKO mice sclerosis, (3) Charcot-Marie-Tooth disease, (4) mentioned with respect to Fig. 1A, compared to con- Waardenburg syndrome, (5) Guillain-Barré syndrome, trol. The IP confirms the result shown in Fig. 1A. (6) chronic inflammatory demyelination polyneuropathy, (7) demyelination due to aging, diabetes or due to toxic Figures 2A, 2B and 2C show WB images of Ace- agents, (8) demyelination and hypomyelination due to 25 eEFIA (presumably Ace-eEF1A1) following IP of other diseases, such as Acute disseminated encephalo- Sox10 or eEF1A1 (Fig. 2A) or JMJD2C (Fig. 2C), or myelitis, transverse myelitis, Leukodystrophy, Central of HDAC2 following IP of eEF1A1 (Fig. 2B), obtained pontine myelinolysis, Glioma, (9) schizophrenia, (10) de- from de-differentiated rat Schwann cells cultured in myelination after traumatic lesion of the CNS. the presence of the HDAC1/2 inhibitor Mocetinostat [0018] In an aspect, the invention provides a method 30 orits vehicle.Nuclear and cytoplasmic fractions were for screening for agents suitable in the treatment and/or separately analysed.