Targeted O-GlcNAc glycoside hydrolase inhibitors to promote myelin regeneration

Abstract

The invention provides an O-GlcNAc glucoside hydrolase (O-GlcNAcase, OGA) inhibitor for promoting remyelination and application of the O-GlcNAc glucoside hydrolase (O-GlcNAcase, OGA) inhibitor. Specifically, the OGA inhibitor with druggability is obtained by comparing the promotion effects of different types of OGA inhibitors on OPC differentiation and myelin regeneration in an oligodendrocyte progenitor cell differentiation model and a myelin regeneration model. The OGA inhibitor provided by the invention has wide application value in prevention and / or treatment of demyelination and related diseases thereof.

Description

Technical Field

[0001] The present invention relates to the field of neurological medicine, and in particular to a targeted O-GlcNAc glycoside hydrolase inhibitor and an application thereof in promoting myelin regeneration. Background Art

[0002] In the central nervous system, oligodendrocyte progenitor cells (OPCs) differentiate into oligodendrocytes (OLs), forming myelin structures that wrap around neuronal axons, which helps to efficiently transmit nerve signals, thereby regulating neural circuits and ensuring the physiological functions of the brain. If OPCs cannot be effectively activated, proliferated, and differentiated into myelinating OLs, it will lead to impaired myelin regeneration, causing demyelinating diseases, nerve death and degenerative diseases.

[0003] The etiology of demyelinating diseases is complex. Currently approved first-line clinical drugs are mostly based on immunomodulation. This treatment method can only delay the progression of the disease and cannot effectively promote myelin regeneration. Clinical data show that there are a certain number of OPCs in the lesion area of ​​patients with multiple sclerosis, but the lack of the ability to differentiate and produce myelin is the main cause of myelin regeneration disorders. Therefore, finding methods to promote the differentiation of OPCs is of great significance for myelin regeneration, the correct establishment and maintenance of brain function, and functional repair and brain function reconstruction after nerve injury. However, the intrinsic determinants that regulate the proliferation and differentiation of OPCs and the targets of drug therapy are still unclear. Therefore, the key to solving demyelinating diseases lies in finding key targets that promote OPC differentiation and myelination, thereby achieving in situ regeneration of myelin.

[0004] Therefore, there is an urgent need in the art to develop drugs that can prevent and / or treat demyelination-related diseases. Summary of the invention

[0005] The object of the present invention is to provide OGA inhibitor compounds that can prevent and / or treat demyelination-related diseases.

[0006] In a first aspect of the present invention, there is provided a use of an O-GlcNAc glycoside hydrolase (O-GlcNAcase, OGA) inhibitor for preparing a preparation or composition, wherein the preparation or composition is used for one or more applications selected from the following group:

[0007] (a) preventing and / or treating demyelination-related diseases;

[0008] (b) promoting the generation, maintenance and / or proliferation of oligodendrocyte precursor cells (OPCs);

[0009] (c) Promote the differentiation of oligodendrocyte progenitor cells (OPC) into oligodendrocytes (OL) and promote the maturation of OL;

[0010] (d) promoting myelination and / or regeneration;

[0011] In another preferred embodiment, the OGA inhibitor includes a compound selected from the following group or a pharmaceutically acceptable salt thereof:

[0012] (1) Compound of formula I:

[0013]

[0014] In the formula,

[0015] X1 is -CH-, or -N-;

[0016] X2 is -CHR a -, where R a is H, or C1-C3 alkyl;

[0017] n is 1 or 2;

[0018] L1 is -(CHR b ) n1 -, -O-, -NH-, or -O-CH2-; wherein R b is H, or C1-C3 alkyl; n1 is an integer of 0-4;

[0019] L2 is -(CHR c ) n2 -, -O-, -NH-, or -O-CH2-; wherein R c is H, or C1-C3 alkyl; n2 is an integer of 0-4;

[0020] Ring A and Ring B are each independently a substituted or unsubstituted 6-10 membered aryl group, or a substituted or unsubstituted 5-10 membered heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S; the substitution refers to substitution by one or more substituents selected from the following group: halogen, C1-C3 alkyl, hydroxyl, C1-C3 alkoxy, C3-C4 cycloalkyl, or two adjacent substituents and the connected ring atoms together form a 5-7 membered carbocyclic ring or a 5-7 membered heterocyclic ring, wherein the 5-7 membered heterocyclic ring contains 1-3 heteroatoms selected from N, O and S;

[0021] W1 and W2 are each independently none, H, D, -CN, -COR1, -(NR d )-CO-R2, substituted or unsubstituted C1-C6 alkyl, or substituted or unsubstituted C3-C5 cycloalkyl; wherein R dR1 and R2 are each independently H, a substituted or unsubstituted group selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, and C3-C6 cycloalkoxy, wherein the substitution refers to substitution by one or more substituents: halogen, C1-C3 alkyl, hydroxy, and phenyl;

[0022] (2) Compound of formula II (PB-001):

[0023]

[0024] In another preferred embodiment, A is a divalent group selected from the following group:

[0025] Wherein, Y1 is -O- or -S-, Y2 is =CH-, =CF- or =N-; Y3 and Y4 are each independently =CR e - or =N-, where R e is H, halogen, C1-C3 alkyl, or C1-C3 haloalkyl; Y5 and Y6 are each independently -O- or -CH2-; Y7 is -N= or -CH=; Y8 is -O-, -S- or -CH2-.

[0026] In another preferred embodiment, B is a divalent group selected from the following group:

[0027] Wherein, Z1, Z2 and Z3 are each independently =CR f - or =N-, at least one of which is =N-; R f is H, or C1-C3 alkyl; Z4 and Z6 are each independently -O- or -CHR g , R g is H, or C1-C3 alkyl; Z5 is N or CH; Z7 and Z8 are each independently =CR h -or=N-,R h It is H, or C1-C3 alkyl.

[0028] In another preferred embodiment, W1 is H, C1-C4 alkyl, or C3-C4 cycloalkyl, wherein R2 is as defined above.

[0029] In another preferred embodiment, W2 is H, -CN, -COR1, C1-C4 alkyl, or C3-C4 cycloalkyl, wherein R1 is as defined above.

[0030] In another preferred embodiment, the OGA inhibitor is a compound selected from Table 1:

[0031] Table 1 Preferred OGA inhibitors

[0032]

[0033]

[0034]

[0035]

[0036] In another preferred embodiment, the OGA inhibitor is selected from the following group: A1-67, PB-001, L1-01, L1-02, L1-03, L1-04, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-01, L4-02, L4-03, L4-04, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06, L5-07, L5-08.

[0037] In another preferred embodiment, the OGA inhibitor is selected from the following group: A1-67, PB-001, L1-01, L1-02, L1-03, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-01, L4-02, L4-03, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06, L5-07.

[0038] In another preferred embodiment, the demyelination-related disease is selected from the following group: multiple sclerosis, neuromyelitis optica, spinal cord injury, demyelination caused by brain injury, demyelination, mental illness and degenerative diseases caused by demyelination, myelin regeneration disorders caused by aging, or a combination thereof.

[0039] In another preferred embodiment, the demyelination includes acute or chronic demyelination.

[0040] In another preferred embodiment, the demyelination includes age-related demyelination and demyelination caused by central nervous system damage.

[0041] In another preferred embodiment, the demyelination includes demyelination caused by external factors such as environment, nutrition, and viral infection.

[0042] In another preferred embodiment, the demyelination includes demyelination caused by intrinsic factors such as genetic factors (including genetic mutations, autoimmunity) and the like.

[0043] In another preferred embodiment, the demyelination includes: demyelination caused by central nervous system trauma.

[0044] In another preferred embodiment, the brain injury includes brain trauma and cerebral stroke.

[0045] In another preferred embodiment, the preparation or composition contains:

[0046] (a) a safe and effective amount of an OGA inhibitor; and

[0047] (b) a pharmaceutically acceptable carrier.

[0048] In another preferred embodiment, the preparation or composition includes: a solid dosage form, a liquid preparation, or a gel preparation.

[0049] In another preferred embodiment, the preparation or composition includes: an oral dosage form or an injection.

[0050] In another preferred embodiment, the preparation or composition includes: tablets, powders, granules, capsules, lyophilized agents, solutions, and syrups.

[0051] In a second aspect of the present invention, a method for promoting differentiation of oligodendrocyte progenitor cells into oligodendrocytes in vitro is provided, wherein oligodendrocyte progenitor cells are cultured in the presence of an OGA inhibitor, thereby promoting differentiation of the oligodendrocyte progenitor cells into oligodendrocytes.

[0052] In another preferred embodiment, the method is non-diagnostic and non-therapeutic.

[0053] In another preferred embodiment, the cells are cells of human and non-human mammals.

[0054] In another preferred embodiment, the OGA inhibitor is selected from the following group: A1-67, PB-001, L1-01, L1-02, L1-03, L1-04, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-01, L4-02, L4-03, L4-04, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06, L5-07, L5-08.

[0055] In another preferred embodiment, the OGA inhibitor is selected from the following group: A1-67, PB-001, L1-01, L1-02, L1-03, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-01, L4-02, L4-03, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06, L5-07.

[0056] In another preferred embodiment, the culture is carried out in an in vitro culture system (or culture medium), and the concentration of the OGA inhibitor is 0.01-100 nM, preferably 0.05-50 nM, and more preferably 0.1-20 nM.

[0057] In a third aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0058] (a1) a safe and effective amount of an OGA inhibitor as the first active ingredient, wherein the OGA inhibitor is selected from the following group: a compound of formula I or a pharmaceutically acceptable salt thereof; a compound of formula II (PB-001), or a combination thereof;

[0059] (a2) a safe and effective amount of an additional drug for treating a demyelination-related disease as a second active ingredient; and

[0060] (b) a pharmaceutically acceptable carrier.

[0061] In another preferred embodiment, the preparation or composition includes: a solid dosage form, a liquid preparation, or a gel preparation.

[0062] In another preferred embodiment, the preparation or composition includes: an oral dosage form or an injection.

[0063] In another preferred embodiment, the additional drug for treating demyelination-related diseases is selected from the following group: Clemastine, Miconazole, Opicinumab, Quetiapine, GSK239512, Olesoxime, Simvastatin, VX15, GNbAC1, Biotin, rHIgM22, Domperidone, Etazolate, U50488, U46619, Panzyga, Benztropine, Clobetasol, Fluticasone, Atropine, Betamethasone, Chlorprothixene, Trifluoperazine 2HCl, FTY720, Ifenprodil, Toremifene, Raloxifene, 9-cis-Retinoicacid, 2,2-Dimethylzymosterol, TASIN-1, Amorolfine, ketoconazole, XAV939, CW3388, Ro25-6981, Ro 31-8220, Fasudil, Emricasan, Perphenazine, Sildenafil, Tadalafil, Roflumilast, Azaperone, Flupentixol, Bifonazole, Butoconazole, Clotrimazole, Fluconazole.

[0064] In a fourth aspect of the present invention, a medicine kit is provided, comprising:

[0065] (i) a first pharmaceutical composition, comprising (a1) a safe and effective amount of an OGA inhibitor as a first active ingredient, wherein the OGA inhibitor is selected from the group consisting of a compound of formula I or a pharmaceutically acceptable salt thereof; a compound of formula II (PB-001), or a combination thereof; and (b1) a pharmaceutically acceptable carrier;

[0066] (ii) a second pharmaceutical composition comprising (a2) a safe and effective amount of an additional drug for treating a demyelination-related disease as a second active ingredient; and (b2) a pharmaceutically acceptable carrier;

[0067] Wherein, the first pharmaceutical composition and the second pharmaceutical composition are independent of each other.

[0068] In a fifth aspect of the present invention, a method for treating a demyelination-related disease is provided, comprising the step of administering a pharmaceutical composition containing a therapeutically effective amount of an OGA inhibitor to a subject in need of treatment.

[0069] In another preferred embodiment, the subject is a human or non-human mammal.

[0070] In another preferred embodiment, the subject is a human.

[0071] In another preferred embodiment, the subject is a patient.

[0072] In another preferred embodiment, the administration includes oral administration or injection administration.

[0073] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 The effects of L1-01 on OPCs differentiation and remyelination are shown. Figure 1 A shows the immunofluorescence results of the effect of L1-01 treatment on OPCs differentiation. The concentration gradient range of L1-01 is marked as shown in the figure. Scale bar, 50 μm. Figure 1 B and Figure 1 C shows the O4 + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 1 D shows the statistical results of relative cell activity of the L1-01 treatment groups at different concentration gradients. Figure 1 E shows representative images of FluoroMyelin staining results of the corpus callosum in different groups, with a scale bar of 200 μm. The dotted line marks the core injury area with high expression of Iba1, and the area of ​​myelin loss and the average fluorescence intensity of FluoroMyelin are quantitatively analyzed in this area. Figure 1 F, 1G show the area of ​​the damaged area ( Figure 1 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 1 G) Quantitative statistical results. Figure 1 H, 1I show the accelerated rotarod ( Figure 1 H) and the uniform rotating rod ( Figure 1 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 1 BD, Figure 1 HI was analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 1 F. Figure 1G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0075] Figure 2 The effects of L1-03 on OPCs differentiation and remyelination are shown. Figure 2 A shows the immunofluorescence results of the effect of L1-03 treatment on OPCs differentiation. The concentration gradient range of L1-03 is marked as shown in the figure. Scale bar, 50 μm. Figure 2 B and Figure 2 C shows the O4 in the L1-03 treatment group at different concentration gradients + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 2 D shows the relative cell activity statistics of the L1-03 treatment groups at different concentration gradients. Figure 2 E shows representative images of FluoroMyelin staining results of the corpus callosum in different groups, with a scale bar of 200 μm. The dotted line marks the core injury area with high expression of Iba1, and the area of ​​myelin loss and the average fluorescence intensity of FluoroMyelin are quantitatively analyzed in this area. Figure 2 F, 2G show the area of ​​the damaged area ( Figure 2 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 2 G) Quantitative statistical results. Figure 2 H, 2I shows the accelerated rotarod ( Figure 2 H) and the uniform rotating rod ( Figure 2 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 2 BD, Figure 2 HI was analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 2 F. Figure 2 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0076] Figure 3 The effects of L2-01 on OPCs differentiation and remyelination are shown. Figure 3 Immunofluorescence results of the effect of L2-01 treatment on OPCs differentiation in A. Scale bar: 50 μm. Figure 3 B. Figure 3 C shows O4 in the L2-01 treatment group + (Figure B), Mbp + (Figure C) Cell percentage statistics. Figure 3D shows the statistical results of relative cell activity in the L2-01 treated group. Figure 3 E shows representative images of FluoroMyelin staining results among different groups, scale bar 200 μm, where the dotted line marked area is the core injury area with high expression of Iba1. Figure 3 F, 3G show the area of ​​the damaged area ( Figure 3 F) and the mean fluorescence intensity (MFI) of FluoroMyelin fluorescence staining ( Figure 3 G) Quantitative statistical results. Figure 3 H, 3I shows the acceleration ( Figure 3 H) and the uniform rotating rod ( Figure 3 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 3 BD, Figure 3 HI graphs were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 3 F. Figure 3 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0077] Figure 4 The effects of L3-01 on OPCs differentiation and remyelination are shown. Figure 4 A is an immunofluorescence image showing the effect of L3-01 treatment on OPCs differentiation. Scale bar: 50 μm. Figure 4 B. Figure 4 C shows O4 in the L3-01 treatment group + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 4 D shows the statistical results of relative cell activity in the L3-01 treated group. Figure 4 E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high expression of Iba1. Figure 4 F, 4G show the area of ​​the damaged area ( Figure 4 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 4 G) Quantitative statistical results. Figure 4 H, 4I shows the acceleration ( Figure 4 H) and the uniform rotating rod ( Figure 4 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 4 BD, Figure 4HI graphs were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 4 F. Figure 4 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0078] Figure 5 The effects of A1-67 on OPCs differentiation and remyelination are shown. Figure 5 A is an immunofluorescence image showing the effect of A1-67 treatment on OPCs differentiation. Scale bar: 50 μm. Figure 5 B. Figure 5 C shows O4 in the A1-67 treatment group + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 5 D shows the statistical results of relative cell activity of the A1-67 treated group. Figure 5 E shows representative images of FluoroMyelin staining results among different groups, scale bar 200 μm, where the dotted line marked area is the core lesion area with high expression of Iba1. Figure 5 F, 5G respectively shows the area of ​​the damaged area ( Figure 5 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 5 G) Quantitative statistical results. Figure 5 H, 5I display acceleration ( Figure 5 H) and the uniform rotating rod ( Figure 5 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 5 BD, Figure 5 HI graphs were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 5 F. Figure 5 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0079] Figure 6 The effects of PB-001 on OPCs differentiation and remyelination are shown. Figure 6 A is an immunofluorescence image showing the effect of PB-001 treatment on OPCs differentiation. Scale bar: 50 μm. Figure 6 B. Figure 6 C shows O4 in the PB-001 treated group + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 6D shows the statistical results of relative cell activity in the PB-001 treated group. Figure 6 E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high expression of Iba1. Figure 6 F, 6G show the area of ​​the damaged area ( Figure 6 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 6 G) Quantitative statistical results. Figure 6 H, 6I display acceleration ( Figure 6 H) and the uniform rotating rod ( Figure 6 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 6 BD, Figure 6 HI graphs were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 6 F. Figure 6 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0080] Figure 7 The effects of L4-03 on OPCs differentiation and remyelination are shown. Figure 7 A shows the immunofluorescence results of the effect of L4-03 treatment on OPCs differentiation. Scale bar: 50 μm. Figure 7 B. Figure 7 C shows O4 in the L4-03 treatment group + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 7 D shows the statistical results of relative cell activity in the L4-03 treated group. Figure 7 E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high expression of Iba1. Figure 7 F, 7G show the area of ​​the damaged area ( Figure 7 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 7 G) Quantitative statistical results. Figure 7 H, 7I display acceleration ( Figure 7 H) and the uniform rotating rod ( Figure 7 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 7 BD, Figure 7HI graphs were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 7 F. Figure 7 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0081] Figure 8 The effects of L5-02 on OPCs differentiation and remyelination are shown. Figure 8 A shows the immunofluorescence results of the effect of L5-02 treatment on OPCs differentiation. Scale bar: 50 μm. Figure 8 B. Figure 8 C shows O4 in the L5-02 treatment group + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 8 D shows the statistical results of relative cell activity in the L5-02 treated group. Figure 8 E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high expression of Iba1. Figure 8 F, 8G show the area of ​​the damaged area ( Figure 8 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 8 G) Quantitative statistical results. Figure 8 H, 8I display acceleration ( Figure 8 H) and the uniform rotating rod ( Figure 8 I) Experimental statistical results. The data are shown as mean ± standard deviation. Each group includes 2 female mice and 2 male mice. Figure 8 BD, Figure 8 HI graphs were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 8 F. Figure 8 G Two-tailed T test was used, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0082] Fig. 9 The effects of different OGA inhibitors on OPCs differentiation and remyelination are shown. Fig. 9 Representative immunofluorescence results showing the effect of adding the OGA inhibitor in this experiment (the optimal concentration in this experiment is marked in the figure) to the OPC culture medium on the differentiation of OPCs. + and Mbp + Cells, scale bar 50 μm. DETAILED DESCRIPTION

[0083] After extensive and in-depth research, the inventors found that O-GlcNAcylation modification can promote myelin generation and / or regeneration, prevent and / or treat diseases related to demyelination, promote oligodendrocyte progenitor cells to differentiate and mature into oligodendrocytes, promote the generation, maintenance and / or proliferation of oligodendrocyte progenitor cells, and OGA inhibitors can increase the level of O-GlcNAcylation modification, thereby significantly enhancing the remyelination of the demyelination model, and can reduce the severity of demyelination, and restore the relevant functions of the body. The inventors also compared a large number of different types of OGA inhibitors in OPCs differentiation model and myelin regeneration model to promote myelin regeneration, and verified the restorative effect of OGA inhibitors on the behavior of demyelination mouse model in vivo, thereby finding that it has great potential application value in promoting myelin regeneration in the treatment of demyelinating diseases. The present invention was completed on this basis.

[0084] OGA inhibitors of the present invention

[0085] As used herein, the terms "O-linked glycosylation hydrolase inhibitor", "O-GlcNAc glycoside hydrolase inhibitor", "OGA inhibitor", "small molecule compound of the present invention" and "compound of the present invention" are used interchangeably and all refer to glycoside hydrolase (OGA) inhibitors that can promote O-linked beta-N-acetylglucosaminylation (O-linked beta-N-acetylglucosaminylation, abbreviated as: O-GlcNAcylation or O-GlcNAc) in the present invention.

[0086] Some preferred OGA inhibitors of the present invention are compounds selected from the group consisting of A1-67, PB-001, L1-01, L1-02, L1-03, L1-04, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-01, L4-02, L4-03, L4-04, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06, L5-07, L5-08.

[0087] Preferably, the compound of the present invention is the OGA inhibitor, or a pharmaceutically acceptable salt thereof, or an optical isomer thereof, or a racemate thereof, or a solvate thereof.

[0088] In the present invention, pharmaceutically acceptable salts of the compounds are also included. The term "pharmaceutically acceptable salt" refers to a salt formed by the compound of the present invention and an acid or a base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by the compound of the present invention and an acid. Acids suitable for forming salts include but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, benzenesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid.

[0089] The compounds of the present invention can be prepared by methods well known to those skilled in the art in the prior art, and there is no particular limitation on the reaction parameters of each step.

[0090] As used herein, in the compounds shown in the above table, if a chiral carbon atom exists, the chiral carbon atom may be in the R configuration, or in the S configuration, or a mixture of the two.

[0091] The promoting effect of the OGA inhibitor of the present invention on myelin regeneration

[0092] The OGA inhibitor of the present invention can increase O-linked beta-N-acetylglucosaminylation (O-linked beta-N-acetylglucosaminylation, referred to as: O-GlcNAcylation or O-GlcNAc). O-GlcNAcylation is a dynamic post-translational modification process catalyzed by a pair of enzymes with opposite effects, namely O-linked glycosyl transferase (OGT) and glycoside hydrolase (OGA). OGT transfers beta-N-acetylglucosamine (O-GlcNAc) to the hydroxyl group of serine / threonine residues of nucleoplasmic proteins, while OGA is responsible for removing the modification. O-GlcNAcylation appears as a metabolic sensor and can participate in the regulation of various biological processes, such as gene expression, signal transduction, protein homeostasis, nutrient perception and cell response to different stress conditions. Among them, O-GlcNAcylation modification is highly enriched in the brain and is essential for the normal development and function of neurons.

[0093] Oligodendrocyte progenitor cells (OPCs) are resident progenitor cells distributed throughout the central nervous system (CNS). Their main function is to form myelin sheaths that wrap around neuronal axons by proliferating and differentiating into oligodendrocytes (OLs), so that neural signals can be quickly transmitted. In addition, OPCs can also ensure neuronal activity and synchronize neuronal circuits by differentiating into OLs, thereby affecting brain function. Aging, gene mutation, nutrition, autoimmune diseases and / or injuries can lead to the destruction of myelin structure and ultimately lead to demyelinating diseases. Although the functional impairment caused by myelin damage can be partially restored by myelin regeneration, the efficiency of myelin regeneration is very low. Clinical studies have shown that OPCs exist in large numbers in demyelinated areas, but cannot differentiate into mature OLs. Therefore, the failure of OPC differentiation is one of the reasons that hinder myelin repair.

[0094] The inventors have found through research that the OGA inhibitor of the present invention can increase the O-GlcNAcylation modification level of OPCs, promote OPCs differentiation, and then promote myelin formation, and achieve myelin regeneration after injury. The OGA inhibitor of the present invention significantly enhances myelin regeneration in a demyelination model, leads to a reversal of the severity of the disease, and restores the body's related functions, and can be used to prevent or treat diseases related to demyelination.

[0095] Pharmaceutical compositions and administration methods of the present invention

[0096] Since the OGA inhibitor of the present invention has an excellent effect of promoting myelination and / or regeneration, the OGA inhibitor of the present invention (including the compound and its various crystal forms, and pharmaceutically acceptable salts) and a pharmaceutical composition containing the OGA inhibitor of the present invention as a main active ingredient can be used to prevent and / or treat (stabilize, alleviate or cure) demyelination-related diseases.

[0097] The pharmaceutical composition of the present invention comprises an OGA inhibitor of the present invention within a safe and effective amount and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition (such as one dose) contains 2mg-1000mg of the OGA inhibitor of the present invention, and more preferably, contains or 10-500mg of the OGA inhibitor of the present invention. Preferably, the "one dose" is a capsule or tablet or an injection. Preferably, the unit dosage form is a dose corresponding to the daily dose.

[0098] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the targeting promoter of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0099] There is no particular limitation on the administration method of the OGA inhibitor or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration or subcutaneous administration).

[0100] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0101] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0102] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0103] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0104] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0105] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0106] The OGA inhibitor of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0107] When administered in combination, the pharmaceutical composition also includes one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds. One or more of the other pharmaceutically acceptable compounds can be administered simultaneously, separately or sequentially with the compound of the present invention.

[0108] When using the pharmaceutical composition, a safe and effective amount of the targeting promoter of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 mg to 1000 mg, preferably 5 mg to 200 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill range of skilled physicians.

[0109] Treatment

[0110] The present invention also provides a method for treating a demyelination-related disease, comprising the step of administering a therapeutically effective amount of an OGA inhibitor to a subject in need of treatment.

[0111] Preferably, the subject includes humans and non-human mammals (rodents, rabbits, monkeys, livestock, dogs, cats, etc.).

[0112] The main advantages of the present invention include:

[0113] (1) The present invention first discovered that OGA inhibitors can increase the O-GlcNAcylation modification level of OPCs, promote the differentiation and maturation of OPCs into OLs, and thus achieve myelin regeneration after injury.

[0114] (2) The present invention provides a group of OGA inhibitors, which have excellent effects in promoting OPC differentiation and promoting myelin regeneration, and can be used to prevent and / or treat demyelination-related diseases, or to prepare drugs for preventing and / or treating demyelination-related diseases.

[0115] (3) The OGA inhibitor of the present invention can significantly promote the generation, maintenance and / or proliferation of oligodendrocyte progenitor cells, and promote the differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes.

[0116] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0117] Equipment and Materials

[0118] Table 2 Some main experimental materials

[0119]

[0120] Example 1: Effect of OGA inhibitor on promoting OPC differentiation

[0121] Primary NPCs (pNPCs) were isolated from the cerebral cortex of embryonic day 12.5 (E12.5) mice as starting cells, and all starting cells for small molecule validation experiments were derived from the same batch of pNPCs. pNPCs were cultured at 2.5×10 4 Pieces / cm 2Inoculate in a Matrigel-coated 24-well plate and culture in pNPC medium (Zhang M, et al. CELL STEM CELL 2016; 18(5): 653-667; Liu C, et al. J MOL CELL BIOL 2019; 11(6): 489-495.) to promote NPC proliferation and state recovery. Then replace the culture medium with OPC medium, and add the small molecule compounds of the present invention to the OPC medium according to the concentration gradient, and this process lasts for 3 days. Then, in OL medium (Zhang M, et al. CELL STEM CELL 2016; 18(5): 653-667; Liu C, et al. J MOL CELL BIOL 2019; 11(6): 489-495.) to induce differentiation of OPCs into OLs, and continuously add small molecule compounds of the present invention in different concentration gradients to the OL medium, and the induction process is 3 days. The drug solvent DMSO (volume ratio v / V=0.1%) was used as a control group to construct a directional differentiation system using pNPCs to OLs. In the present invention, quantitative analysis was performed by detecting the percentage of cells expressing oligodendrocyte marker genes O4 and Mbp in differentiated cells. O4 and Mbp are both marker genes for mature oligodendrocytes and are recognized in the field as cell markers for measuring the degree of OPC differentiation and myelination. The expression of O4 and Mbp indicates the differentiation and myelination of OPC. Therefore, by detecting cells expressing O4 and Mbp after treatment with different concentration gradients of OGA inhibitors, its effect on OPC differentiation and myelination can be indicated.

[0122] The results are as follows Figure 1-9 As shown in Tables 3 and 4. Tables 3 and 4 respectively show the O4 + (Table 3), Mbp + (Table 4) Statistical results of cell percentage.

[0123] exist Figure 1-8 B, C, Fig. 9 In Tables 3 and 4, the O4 + , Mbp + The percentage of cells was 10%, and the change in cell activity at this concentration was within 10% compared with the control group ( Figure 1 D- Figure 8 D), no obvious cell death was observed.

[0124] Tables 3 and 4 show that different small molecule compounds induce OPCs differentiation after O4 + , Mbp +The percentage of cells was statistically analyzed and recorded compared with the control group (DMSO group). The change in cell activity at this concentration was within 10% compared with the control group, and no obvious cell death occurred. Based on the above standards, the effective concentration range of each small molecule was determined, as shown in Table 3 and Table 4.

[0125] Table 3 O4 under the action of different compounds + Cell percentage statistics

[0126]

[0127]

[0128] Note: *** indicates that when the concentration is 0.1nM, O4 + Cell percentage ≥ 20%; ** indicates that when the concentration is 0.1nM, O4 + When the cell percentage is less than 20%, the concentration is 0.5nM, O4 + Cell percentage ≥20%; * indicates others.

[0129] Table 4 Mbp under the action of different compounds + Cell percentage statistics

[0130]

[0131]

[0132] Note: *** indicates that when the concentration is 0.1nM, Mbp + Cell percentage ≥ 13%; ** indicates that when the concentration is 0.1nM, Mbp + The percentage of cells was less than 13%, and when the concentration was 0.5 nM, Mbp + Cell percentage ≥13%; * indicates others.

[0133] The above results show that the small molecule compounds of the present invention can significantly increase O4 + Cell percentage and / or Mbp + The maximum cell percentage, especially compounds A1-67, L1-01, L1-02, L1-03, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-03, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06 at very low concentrations (such as 0.1-5nM) can significantly increase O4 +Cell percentage, while compounds A1-67, L1-01, L1-02, L1-03, L2-01, L2-02, L2-03, L2-04, L3-01, L3-02, L3-03, L3-04, L4-01, L4-03, L5-01, L5-02, L5-03, L5-04, L5-05, L5-06, and L5-07 can significantly increase Mbp at very low concentrations (such as 0.1-5 nM). + Cell percentage. Due to O4 + Cell percentage and / or Mbp + The increase in cell percentage represents the differentiation of oligodendrocyte progenitor cells into oligodendrocytes. Therefore, the above experimental data show that the compound of the present invention can effectively promote the differentiation of oligodendrocyte progenitor cells into oligodendrocytes.

[0134] Example 2: OGA inhibitor promotes myelin regeneration in a demyelinated mouse model

[0135] To investigate whether OGA inhibitors can promote remyelination in vivo, a toxin-induced model was used to induce demyelination in the corpus callosum (CC) region of mice by local injection of lysolecithin (lysophosphatidylcholine, LPC).

[0136] Using a toxin-induced model, mice in this model were subjected to acute demyelinating symptoms by targeted injection of lysophosphatidylcholine (LPC) in the corpus callosum. Demyelination symptoms appeared 4 days after injection, and remyelination began to appear 14 days later. Therefore, the therapeutic effect of OGA inhibitors on remyelination in the LPC model was evaluated in the time window of 4-14 days after injection. This model is a recognized animal model in the field for evaluating remyelination. Mice were continuously administered daily from 4 to 10 days after LPC injection by intraperitoneal injection. The WT+saline group was used as a blank control, and the LPC+saline group was used as a negative control. On the 14th day, the myelin marked by FluoroMyelin and the area size of the demyelinated area marked by the inflammatory area with high expression of Iba1 were used to evaluate the remyelination of the damaged area in the corpus callosum ( Figure 1-Figure 8 , Table 5). The number of animals in each group was 2 female mice and 2 male mice. The statistical method used the two-tailed T test method, and the significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0137] FluoroMyelin can quickly and selectively label the myelin in brain slices, and its fluorescence intensity can be used as an indicator to measure the level of myelin regeneration. Therefore, FluoroMyelin can be used to indicate the effect of OGA inhibitors on myelin regeneration in demyelinated areas.

[0138] Through staining, it was found that the control group of LPC mouse model injected with normal saline showed severe myelin demyelination. The area of ​​myelin demyelination in the corpus callosum lesion area and the average level after FluoroMyelin fluorescence staining were quantitatively analyzed. Figure 1-8 , as shown in Table 5.

[0139] exist Figure 1-8 In the EF, the myelin damage area of ​​the LPC+saline group and the experimental group treated with small molecules were quantitatively analyzed. The myelin damage area was reduced after drug treatment; and the myelin fluorescence staining analysis of FluoroMyelin showed that compared with the control group, the average fluorescence intensity of FluoroMyelin in the corpus callosum damage area of ​​the drug treatment group was significantly increased relative to the LPC damage group. The summary data are shown in Table 5.

[0140] Figure 1-8 F and G show the area of ​​myelin damage ( Figure 1 F) and FluoroMyelin mean fluorescence intensity (MFI) ( Figure 1 G) Quantitative statistical results.

[0141] Table 5 Myelin regeneration in mouse LPC model

[0142]

[0143] In summary Figure 1-8 ,The results in Table 5 show that A1-67, PB-001, L1-01, L1-03, L2-01, L3-01, L4-03, and L5-02 all promote myelin regeneration in the LPC model.

[0144] Example 3: Effect of OGA inhibitor on behavioral recovery of demyelination mouse model

[0145] To further evaluate whether OGA inhibitors can promote the behavioral recovery of the LPC demyelinating disease mouse model, the motor coordination and limb movement ability of the LPC model mice were evaluated using the accelerating rotarod test and the constant speed rotarod test.

[0146] In the accelerated rotating rod experiment, the initial speed was 4 rpm / min and the acceleration was 20 rpm / min. 2The final speed was 40 rpm / min, and the maximum statistical time was 5 min. In the constant speed rotarod experiment, the speed was 20 rpm / min, and the maximum statistical time was 1 min. The effect of OGA inhibitors on the motor ability of demyelinated mice was evaluated by recording the time the mice maintained movement on the accelerating rotarod and the constant speed rotarod.

[0147] In the accelerated experiment, to verify the therapeutic effects of L1-01, L1-03, L2-01, L3-01, A1-67, PB-001, L4-03, and L5-02, LPC mice were treated with L1-01 (900 μg / kg), L1-03 (900 μg / kg), L2-01 (900 μg / kg), L3-01 (900 μg / kg), A1-67 (4500 μg / kg), PB-001 (40 mg / kg), L4-03 (900 μg / kg), or L5-02 (900 μg / kg) daily from day 4 to 10 after model establishment, and behavioral scores were evaluated on day 14. The number of animals in each group was 2 female mice and 2 male mice. The statistical method used was one-way analysis of variance (ANOVA) and post-hoc Tukey's test. The significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0148] The results showed that after drug treatment, the time mice stayed on the rod was significantly increased. The data are shown in Table 6 and Figure 1-8 H.

[0149] In the uniform speed experiment, to verify the therapeutic effects of L1-01, L1-03, L2-01, L3-01, A1-67, PB-001, L4-03, and L5-02, LPC mice were treated with L1-01 (900 μg / kg), L1-03 (900 μg / kg), L2-01 (900 μg / kg), L3-01 (900 μg / kg), A1-67 (4500 μg / kg), PB-001 (40 mg / kg), L4-03 (900 μg / kg), or L5-02 (900 μg / kg) daily from day 4 to 10 after model establishment, and behavioral scores were evaluated on day 14. The number of animals in each group was 2 female mice and 2 male mice. The statistical method used was one-way analysis of variance (ANOVA) and post-hoc Tukey's test. The significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0150] The results showed that the time mice stayed on the rod was significantly increased after being treated with L1-01, L1-03, L2-01, L3-01, A1-67, PB-001, L4-03, and L5-02, respectively. The data are shown in Table 6 and Figure 1-8 I.

[0151] Table 6 The time mice remained on the rod

[0152]

[0153] The above results show that the compounds of the present invention can significantly improve the impaired motor ability of demyelination model mice.

[0154] discuss

[0155] O-linked beta-N-acetylglucosamine glycosylation (O-GlcNAcylation, O-GlcNAc) is a protein post-translational modification (referred to as "glycosylation") that is highly enriched in brain tissue, and its level is downregulated with aging. O-GlcNAcylation is catalyzed by glycosyltransferase (O-GlcNAc transferase, Ogt, responsible for glycosylation) and glycoside hydrolase (O-GlcNAcase, Oga, responsible for deglycosylation). Glycosylation can regulate a variety of biological processes such as cell proliferation, migration, and differentiation by regulating protein phosphorylation modification, protein stability, protein localization, protein-protein interactions, and gene expression. Studies have shown that glycosylation is involved in the functional maintenance and homeostatic regulation of the nervous system. For example, the downregulation of O-GlcNAcylation modification levels in neural stem cells (NSCs) affects the cell fate determination of NSCs in the hippocampus and impairs the body's cognitive function. However, whether O-GlcNAcylation modification plays a role in the (re)myelination of the central nervous system has not yet been reported.

[0156] At present, the chemical drugs that promote O-GlcNAcylation modification are mainly inhibitors of glycoside hydrolases (OGA). In the field of tumor research, abnormal O-GlcNAcylation modification in cells is directly related to the growth and proliferation, immune escape and tumor metastasis of various tumor cells, such as leukemia, cervical cancer, colorectal cancer, etc. In addition, OGA inhibitors have begun to be used clinically to affect the proliferation of tumor cells by promoting O-GlcNAcylation modification of cells, thereby inhibiting tumor growth. However, the application of OGA inhibitors in the treatment of demyelination-related diseases has not been studied.

[0157] The root cause of myelination disorders is that OPCs cannot differentiate into OLs with myelination function. In the present invention, by evaluating the effects of a series of OGA inhibitors on promoting myelin regeneration, it was unexpectedly found that certain OGA inhibitors can effectively promote OPC differentiation and optimize the optimal concentration; it was confirmed by an animal demyelination model that OGA inhibitors can promote myelin regeneration and promote behavioral recovery in demyelinated mouse models. A possible mechanism of action of the OGA inhibitor of the present invention is to promote the differentiation ability of OPCs by increasing the O-GlcNAcylation modification level of OPCs, thereby promoting the occurrence process of the oligodendrocyte lineage and achieving myelin regeneration after injury. The present invention will contribute to the research and development of drugs for demyelination and its related diseases, neurodegenerative diseases, and brain aging-related diseases.

[0158] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. Use of an O-GlcNAc glycoside hydrolase (O-GlcNAcase, OGA) inhibitor, characterized in that, For preparing a preparation or composition for one or more applications selected from the following group: (a) preventing and / or treating demyelination-related diseases; (b) promoting the generation, maintenance and / or proliferation of oligodendrocyte precursor cells (OPCs); (c) Promote the differentiation of oligodendrocyte progenitor cells (OPC) into oligodendrocytes (OL) and promote the maturation of OL; (d) Promote myelination and / or regeneration.

2. The use according to claim 1, characterized in that The OGA inhibitor includes a compound selected from the following group or a pharmaceutically acceptable salt thereof: (1) Compound of formula I: In the formula, X1 is -CH-, or -N-; X2 is -CHR a -, where R a is H, or C1-C3 alkyl; n is 1 or 2; L1 is -(CHR b ) n1 -, -O-, -NH-, or -O-CH2-; wherein R b is H, or C1-C3 alkyl; n1 is an integer of 0-4; L2 is -(CHR c ) n2 -, -O-, -NH-, or -O-CH2-; wherein R c is H, or C1-C3 alkyl; n2 is an integer of 0-4; Ring A and Ring B are each independently a substituted or unsubstituted 6-10 membered aryl group, or a substituted or unsubstituted 5-10 membered heteroaryl group, wherein the heteroaryl group contains 1-3 heteroatoms selected from N, O and S; the substitution refers to substitution by one or more substituents selected from the following group: halogen, C1-C3 alkyl, hydroxyl, C1-C3 alkoxy, C3-C4 cycloalkyl, or two adjacent substituents and the connected ring atoms together form a 5-7 membered carbocyclic ring or a 5-7 membered heterocyclic ring, wherein the 5-7 membered heterocyclic ring contains 1-3 heteroatoms selected from N, O and S; W1 and W2 are each independently none, H, D, -CN, -COR1, -(NR d )-CO-R2, substituted or unsubstituted C1-C6 alkyl, or substituted or unsubstituted C3-C5 cycloalkyl; wherein R d R1 and R2 are each independently H, a substituted or unsubstituted group selected from the group consisting of C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, and C3-C6 cycloalkoxy, wherein the substitution refers to substitution by one or more substituents: halogen, C1-C3 alkyl, hydroxy, and phenyl; (2) Compound of formula II (PB-001):

3. The use according to claim 1, characterized in that A is a divalent group selected from the following group: Wherein, Y1 is -O- or -S-, Y2 is =CH-, =CF- or =N-; Y3 and Y4 are each independently =CR e - or =N-, where R e is H, halogen, C1-C3 alkyl, or C1-C3 haloalkyl; Y5 and Y6 are each independently -O- or -CH2-; Y7 is -N= or -CH=; Y8 is -O-, -S- or -CH2-.

4. The use according to claim 1, characterized in that B is a divalent group selected from the following group: Wherein, Z1, Z2 and Z3 are each independently =CR f - or =N-, at least one of which is =N-; R f is H, or C1-C3 alkyl; Z4 and Z6 are each independently -O- or -CHR g , R g is H, or C1-C3 alkyl; Z5 is N or CH; Z7 and Z8 are each independently =CR h -or=N-,R h It is H, or C1-C3 alkyl.

5. The use according to claim 1, characterized in that The OGA inhibitor is a compound selected from the group consisting of:

6. The use according to claim 1, characterized in that The demyelination-related disease is selected from the following group: multiple sclerosis, neuromyelitis optica, spinal cord injury, demyelination caused by brain injury, demyelination, mental illness and degenerative diseases caused by demyelination, myelin regeneration disorder caused by aging, or a combination thereof.

7. The use according to claim 1, characterized in that The preparation or composition contains: (a) a safe and effective amount of an OGA inhibitor; and (b) a pharmaceutically acceptable carrier.

8. A method for promoting differentiation of oligodendrocyte progenitor cells into oligodendrocytes in vitro, characterized in that: Oligodendrocyte progenitor cells are cultured in the presence of an OGA inhibitor, thereby promoting differentiation of the oligodendrocyte progenitor cells into oligodendrocytes.

9. A pharmaceutical composition, characterized in that The pharmaceutical composition contains: (a1) a safe and effective amount of an OGA inhibitor as the first active ingredient, wherein the OGA inhibitor is selected from the following group: a compound of formula I or a pharmaceutically acceptable salt thereof; a compound of formula II (PB-001), or a combination thereof; (a2) a safe and effective amount of an additional drug for treating a demyelination-related disease as a second active ingredient; and (b) a pharmaceutically acceptable carrier.

10. A medicine box, characterized in that: The medicine kit comprises: (i) a first pharmaceutical composition, comprising (a1) a safe and effective amount of an OGA inhibitor as a first active ingredient, wherein the OGA inhibitor is selected from the group consisting of a compound of formula I or a pharmaceutically acceptable salt thereof; a compound of formula II (PB-001), or a combination thereof; and (b1) a pharmaceutically acceptable carrier; (ii) a second pharmaceutical composition comprising (a2) a safe and effective amount of an additional drug for treating a demyelination-related disease as a second active ingredient; and (b2) a pharmaceutically acceptable carrier; Wherein, the first pharmaceutical composition and the second pharmaceutical composition are independent of each other.

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