O-glcnacase-targeting inhibitor for promoting remyelination

Abstract

An O-GlcNAcase (OGA) inhibitor for promoting remyelination and use thereof. Specifically, the OGA inhibitor with druggability is obtained by comparing the promoting effects of different types of OGA inhibitors on oligodendrocyte precursor / progenitor cell differentiation and remyelination in an OPC differentiation model and a remyelination model. The OGA inhibitor has wide application value in preventing and / or treating demyelination and related diseases thereof.

Description

Targeting O-GlcNAc glycoside hydrolase inhibitors to promote myelin regeneration 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 its application in promoting myelin regeneration. Background Art

[0002] In the central nervous system, oligodendrocyte precursor / progenitor cells (OPCs) differentiate into oligodendrocytes (OLs), forming the myelin sheath that wraps around neuronal axons, facilitating the efficient conduction of neural signals, thereby regulating neural circuits and ensuring the physiological functions of the brain. If OPCs cannot effectively activate, proliferate, and differentiate into myelinating OLs, myelin regeneration is impaired, leading to demyelinating diseases, neuronal death, and degenerative diseases.

[0003] The causes of demyelinating diseases are 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 ​​multiple sclerosis patients, but the lack of the ability to differentiate and produce myelin is the main cause of myelin regeneration disorders. Therefore, finding methods to promote OPC differentiation is of great significance for myelin regeneration, the correct establishment and maintenance of brain function, as well as functional repair and brain function reconstruction after nerve injury. However, the intrinsic determinants that regulate OPC proliferation and differentiation and the targets of drug treatment are still unclear. Therefore, the key to solving demyelinating diseases lies in finding key targets that promote OPC differentiation and myelination, so as to achieve 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.

[0005] Summary of the Invention

[0006] The object of the present invention is to provide an OGA inhibitor compound that can prevent and / or treat demyelination and its related diseases.

[0007] 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 formulation or composition for one or more applications selected from the group consisting of:

[0008] (a) Preventing and / or treating demyelination and its related diseases;

[0009] (b) promoting the generation, maintenance and / or proliferation of oligodendrocyte precursor / progenitor cells (OPCs);

[0010] (c) Promote the differentiation of oligodendrocyte progenitor cells (OPCs) into oligodendrocytes (OLs) and promote OL maturation;

[0011] (d) promoting myelination and / or regeneration;

[0012] In another preferred embodiment, the OGA inhibitor includes a compound selected from the following group or a pharmaceutically acceptable salt thereof:

[0013] (1) Compound of formula I:

[0014] Where,

[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 from 0 to 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 from 0 to 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 with one or more substituents selected from the group consisting of halogen, C1-C3 alkyl, hydroxy, 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 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;

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

[0023] In another preferred embodiment, the OGA inhibitor includes a compound selected from the following group or a pharmaceutically acceptable salt thereof:

[0024] (3) Compound represented by formula III:

[0025] in,

[0026] X5 is -N- or -CH-;

[0027] X6 is -N=, -O- or -S-;

[0028] W3 is H, F, -R2- or -OR 2 , R 2 is zero, substituted or unsubstituted C1-C8 alkyl;

[0029] Ring C is 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 group consisting of halogen, C1-C3 alkyl, hydroxy, 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;

[0030] (4) Compound represented by formula IV:

[0031] In another preferred embodiment, A is a divalent group selected from the following group:

[0032] wherein Y1 is -O- or -S-, Y2 is =CH-, =CF- or =N-; Y3 and Y4 are each independently =CR e - or =N-, where R eis 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-.

[0033] In another preferred embodiment, B is a divalent group selected from the following group:

[0034] 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 is H, or C1-C3 alkyl.

[0035] In another preferred embodiment, W1 is H, C1-C4 alkyl, or C3-C4 cycloalkyl, wherein R2 is as defined above.

[0036] In another preferred embodiment, W2 is H, -CN, -COR1, C1-C4 alkyl, or C3-C4 cycloalkyl, wherein R1 is as defined above.

[0037] In another preferred embodiment, the OGA inhibitor is a compound selected from Table 1:

[0038] Table 1 Preferred OGA inhibitors

[0039] In another preferred embodiment, the OGA inhibitor is a compound selected from Table B:

[0040] Table B Preferred OGA inhibitors

[0041] In another preferred embodiment, the OGA inhibitor is 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, L5-09, L5-10 7. L5-08, A1-01, AT1-01, AT1-02, B1-01, B1-02, B2-1-01, B2-1-02, B2-2-01, B 2-2-02, B5-01, B5-02, A2-01, A2-02, A3-01, A3-02, A5-01, A5-02, J-01, TP-05.

[0042] 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, J-01, TP-05.

[0043] 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, J-01, TP-05.

[0044] In another preferred embodiment, the demyelination and its related diseases are selected from the following groups: multiple sclerosis, neuromyelitis optica, spinal cord injury, demyelination caused by brain injury, demyelination, psychiatric diseases caused by demyelination, degenerative diseases and / or neurological dysfunction, myelin regeneration disorders caused by aging, or a combination thereof.

[0045] In another preferred embodiment, the demyelination includes acute or chronic demyelination.

[0046] In another preferred embodiment, the demyelination includes age-related demyelination and demyelination caused by central nervous system damage.

[0047] In another preferred embodiment, the demyelination includes demyelination caused by external factors such as environment, nutrition, and viral infection.

[0048] In another preferred embodiment, the demyelination includes demyelination caused by intrinsic factors such as genetic factors (including genetic mutations and autoimmunity).

[0049] In another preferred embodiment, the demyelination includes demyelination caused by central nervous system trauma.

[0050] In another preferred embodiment, the brain injury includes brain trauma and stroke.

[0051] In another preferred embodiment, the preparation or composition contains:

[0052] (a) a safe and effective amount of an OGA inhibitor; and

[0053] (b) a pharmaceutically acceptable carrier.

[0054] In another preferred embodiment, the preparation or composition includes: a solid dosage form, a liquid preparation, or a gel preparation.

[0055] In another preferred embodiment, the preparation or composition includes: an oral dosage form or an injection.

[0056] In another preferred embodiment, the preparation or composition includes: tablets, powders, granules, capsules, lyophilized agents, solutions, and syrups.

[0057] In a second aspect of the present invention, a method for promoting the 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 the differentiation of the oligodendrocyte progenitor cells into oligodendrocytes.

[0058] In another preferred embodiment, the method is non-diagnostic and non-therapeutic.

[0059] In another preferred embodiment, the cells are cells of human and non-human mammals.

[0060] 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, A1-01, AT1-01, AT1-02, B1-01, B1-02, B2-1-01, B2-1-02 , B2-2-01, B2-2-02, B5-01, B5-02, A2-01, A2-02, A3-01, A3-02, A5-01, A5-02, J-01, TP-05.

[0061] 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, J-01, TP-05.

[0062] 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, J-01, TP-05.

[0063] 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.

[0064] In a third aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0065] (a1) a safe and effective amount of an OGA inhibitor as the 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;

[0066] (a2) a safe and effective amount of an additional drug for treating a demyelination-related disease as a second active ingredient; and

[0067] (b) a pharmaceutically acceptable carrier.

[0068] In another preferred embodiment, the OGA inhibitor is selected from the following group: a compound of formula III or a pharmaceutically acceptable salt thereof; a compound of formula IV (TP-05), or a combination thereof.

[0069] In another preferred embodiment, the preparation or composition includes: a solid dosage form, a liquid preparation, or a gel preparation.

[0070] In another preferred embodiment, the preparation or composition includes: an oral dosage form or an injection.

[0071] 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-Retinoic acid, 2,2-Dimethylzymosterol, TASIN-1, Amorolfine, ketoconazole, XAV939, CW3388, Ro 25-6981, Ro 31-8220, Fasudil, Emricasan, Perphenazine, Sildenafil, Tadalafil, Roflumilast, Azaperone, Flupentixol, Bifonazole, Butoconazole, Clotrimazole, Fluconazole.

[0072] In a fourth aspect of the present invention, a medicine kit is provided, comprising:

[0073] (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;

[0074] (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;

[0075] Wherein, the first pharmaceutical composition and the second pharmaceutical composition are independent of each other.

[0076] In another preferred embodiment, the OGA inhibitor is selected from the following group: a compound of formula III or a pharmaceutically acceptable salt thereof; a compound of formula IV (TP-05), or a combination thereof.

[0077] 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.

[0078] In another preferred embodiment, the subject is a human or non-human mammal.

[0079] In another preferred embodiment, the subject is a human.

[0080] In another preferred embodiment, the subject is a patient.

[0081] In another preferred embodiment, the administration includes oral administration or injection administration.

[0082] 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 described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0083] Figure 1 shows the effects of L1-01 on OPC differentiation and remyelination. Figure 1A shows the immunofluorescence results of the effects of L1-01 treatment on OPC differentiation. The concentration gradient of L1-01 is marked in the figure. Scale bar, 50 μm. Figures 1B and 1C show the O4 + (Figure B), Mbp +(Figure C) Statistical results of cell percentage. Figure 1D shows the statistical results of relative cell activity in the L1-01 treatment group with different concentration gradients. Figure 1E shows representative images of the FluoroMyelin staining results of the corpus callosum between different groups, with a scale of 200 μm. The dotted line marks the core injury area where Iba1 is highly expressed, and the area of ​​myelin loss and the average fluorescence intensity of FluoroMyelin in this area are quantitatively analyzed. Figures 1F and 1G show the quantitative statistical results for the area of ​​the injury area (Figure 1F) and the average fluorescence intensity (MFI) of FluoroMyelin (Figure 1G), respectively. Figures 1H and 1I show the statistical results of the accelerating rotarod (Figure 1H) and the uniform rotarod (Figure 1I) experiments. The data are shown as mean ± standard deviation. There were 2 female mice and 2 male mice in each group. Figures 1B-D and 1H-I were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figure 1F and 1G were analyzed using a two-tailed T test, where significance was defined as *P<0.05, **P<0.01, and ***P<0.001.

[0084] Figure 2 shows the effects of L1-03 on OPC differentiation and remyelination. Figure 2A shows the immunofluorescence results of the effects of L1-03 treatment on OPC differentiation. The concentration gradient of L1-03 is marked in the figure. Scale bar, 50 μm. Figures 2B and 2C show the effects of L1-03 treatment on O4 in the OPC differentiation and remyelination groups. + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 2D shows the statistical results of relative cell activity in the L1-03 treatment group with different concentration gradients. Figure 2E shows representative images of the FluoroMyelin staining results of the corpus callosum between different groups, with a scale of 200 μm. The dotted line marks the core injury area where Iba1 is highly expressed, and the area of ​​myelin loss and the average fluorescence intensity of FluoroMyelin in this area are quantitatively analyzed. Figures 2F and 2G show the quantitative statistical results for the area of ​​the injury area (Figure 2F) and the average fluorescence intensity (MFI) of FluoroMyelin (Figure 2G), respectively. Figures 2H and 2I show the statistical results of the accelerated rotating rod (Figure 2H) and uniform rotating rod (Figure 2I) experiments. The data are shown as mean ± standard deviation. There are n=2 female mice and n=2 male mice in each group. Figures 2B-D and 2H-I were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figures 2F and 2G were analyzed using a two-tailed T test, where significance was defined as *P<0.05, **P<0.01, and ***P<0.001.

[0085] Figure 3 shows the effects of L2-01 on OPC differentiation and remyelination. Figure 3A shows the immunofluorescence results of the effects of L2-01 treatment on OPC differentiation. Scale bar 50 μm. Figures 3B and 3C show the O4 + (Figure B), Mbp + (Panel C) Cell percentage statistics. Figure 3D shows the relative cell viability statistics of the L2-01-treated group. Figure 3E shows representative images of FluoroMyelin staining results between different groups. Scale bar: 200 μm. The dotted line marks the core lesion area with high Iba1 expression. Figures 3F and 3G show the quantitative statistical results of the lesion area (Figure 3F) and the mean fluorescence intensity (MFI) of FluoroMyelin fluorescence staining (Figure 3G), respectively. Figures 3H and 3I show the statistical results of the accelerated (Figure 3H) and constant-speed rotarod (Figure 3I) experiments. Data are presented as mean ± standard deviation. n = 2 female mice and n = 2 male mice per group. Figures 3B-D and 3H-I were analyzed using one-way analysis of variance (ANOVA) with post-hoc Tukey's test; Figures 3F and 3G were analyzed using two-tailed T-tests. Significance is defined as *P < 0.05, **P < 0.01, ***P < 0.001.

[0086] Figure 4 shows the effect of L3-01 on OPC differentiation and remyelination. Figure 4A shows the immunofluorescence results of the effect of L3-01 treatment on OPC differentiation. Scale bar 50 μm. Figures 4B and 4C show the O4 + (Figure B), Mbp + (Panel C) Statistical results of cell percentage. Figure 4D shows the relative cell viability of the L3-01-treated group. Figure 4E shows representative images of FluoroMyelin staining results. Scale bar: 200 μm. The dotted line marks the core lesion area with high Iba1 expression. Figures 4F and 4G show the quantitative statistical results for the lesion area (Figure 4F) and the mean fluorescence intensity (MFI) of FluoroMyelin (Figure 4G), respectively. Figures 4H and 4I show the statistical results of the accelerated (Figure 4H) and constant-speed rotarod (Figure 4I) experiments. Data are presented as mean ± standard deviation. n = 2 female mice and n = 2 male mice per group. Figures 4B-D and 4H-I were analyzed using one-way analysis of variance (ANOVA) with post-hoc Tukey's test; Figures 4F and 4G were analyzed using two-tailed T-tests. Significance is defined as *P < 0.05, **P < 0.01, ***P < 0.001.

[0087] Figure 5 shows the effects of A1-67 on OPC differentiation and remyelination. Figure 5A shows the immunofluorescence results of the effects of A1-67 treatment on OPC differentiation. Scale bar 50 μm. Figures 5B and 5C show the O4 + (Figure B), Mbp + (Panel C) Statistical results of cell percentage. Figure 5D shows the statistical results of relative cell viability in the A1-67-treated group. Figure 5E shows representative images of FluoroMyelin staining results between different groups. Scale bar: 200 μm. The dotted line marks the core lesion area with high Iba1 expression. Figures 5F and 5G show the quantitative statistical results for the lesion area (Figure 5F) and the mean fluorescence intensity (MFI) of FluoroMyelin (Figure 5G), respectively. Figures 5H and 5I show the statistical results of the accelerated (Figure 5H) and constant-speed rotarod (Figure 5I) experiments. Data are shown as mean ± standard deviation. n = 2 female mice and n = 2 male mice per group. Figures 5B-D and 5H-I were analyzed using one-way analysis of variance (ANOVA) with post-hoc Tukey's test; Figures 5F and 5G were analyzed using two-tailed T-tests. Significance is defined as *P < 0.05, **P < 0.01, ***P < 0.001.

[0088] Figure 6 shows the effects of PB-001 on OPC differentiation and remyelination. Figure 6A shows the immunofluorescence results of the effects of PB-001 treatment on OPC differentiation. Scale bar 50 μm. Figures 6B and 6C show the O4 + (Figure B), Mbp + (Panel C) Statistical results of cell percentage. Figure 6D shows the statistical results of relative cell viability in the PB-001-treated group. Figure 6E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high Iba1 expression. Figures 6F and 6G show the quantitative statistical results for the lesion area (Figure 6F) and the mean fluorescence intensity (MFI) of FluoroMyelin (Figure 6G), respectively. Figures 6H and 6I show the statistical results of the accelerated (Figure 6H) and constant-speed rotarod (Figure 6I) experiments. Data are shown as mean ± standard deviation. n = 2 female mice per group, n = 2 male mice. Figures 6B-D and 6H-I were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figures 6F and 6G were analyzed using two-tailed T-tests, where significance is defined as *P < 0.05, **P < 0.01, ***P < 0.001.

[0089] Figure 7 shows the effect of L4-03 on OPC differentiation and remyelination. Figure 7A shows the immunofluorescence results of the effect of L4-03 treatment on OPC differentiation. Scale bar 50 μm. Figures 7B and 7C show the O4 + (Figure B), Mbp + (Panel C) Statistical results of cell percentage. Figure 7D shows the statistical results of relative cell viability in the L4-03-treated group. Figure 7E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high Iba1 expression. Figures 7F and 7G show the quantitative statistical results for the lesion area (Figure 7F) and the mean fluorescence intensity (MFI) of FluoroMyelin (Figure 7G), respectively. Figures 7H and 7I show the statistical results of the accelerated (Figure 7H) and constant-speed rotarod (Figure 7I) experiments. Data are shown as mean ± standard deviation. n = 2 female mice and n = 2 male mice per group. Figures 7B-D and 7H-I were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figures 7F and 7G were analyzed using two-tailed T-tests, where significance is defined as *P < 0.05, **P < 0.01, ***P < 0.001.

[0090] Figure 8 shows the effect of L5-02 on OPC differentiation and remyelination. Figure 8A shows the immunofluorescence results of the effect of L5-02 treatment on OPC differentiation. Scale bar 50 μm. Figures 8B and 8C show the O4 + (Figure B), Mbp + (Figure C) Statistical results of cell percentage. Figure 8D shows the statistical results of relative cell viability in the L5-02-treated group. Figure 8E shows representative images of FluoroMyelin staining results, scale bar 200 μm, where the dotted line marks the core lesion area with high Iba1 expression. Figures 8F and 8G show the quantitative statistical results for the lesion area (Figure 8F) and the mean fluorescence intensity (MFI) of FluoroMyelin (Figure 8G), respectively. Figures 8H and 8I show the statistical results of the accelerated (Figure 8H) and constant-speed rotarod (Figure 8I) experiments. Data are shown as mean ± standard deviation. n = 2 female mice and n = 2 male mice per group. Figures 8B-D and 8H-I were analyzed using one-way analysis of variance (ANOVA) and post-hoc Tukey's test; Figures 8F and 8G were analyzed using two-tailed T-tests, where significance is defined as *P < 0.05, **P < 0.01, ***P < 0.001.

[0091] Figure 9 shows the effects of different OGA inhibitors on OPC differentiation and remyelination. Figure 9 shows representative immunofluorescence results of the effects of adding OGA inhibitors in this experiment (as marked in the figure, the optimal concentration in this experiment) on OPC differentiation in OPC culture medium, detecting O4 + and Mbp + Cells, scale bar 50 μm. DETAILED DESCRIPTION

[0092] After extensive and in-depth research, the inventors found that O-GlcNAcylation modification can promote myelination and / or regeneration, prevent and / or treat demyelination and its related diseases, promote oligodendrocyte progenitor cells to oligodendrocyte differentiation and maturation, promote the generation, maintenance and / or proliferation of oligodendrocyte progenitor cells, OGA inhibitors can improve O-GlcNAcylation modification levels, thereby significantly enhancing the remyelination of demyelination models, and can alleviate the severity of demyelination, restore the relevant functions of the body. The inventors also compared a large number of different types of OGA inhibitors in OPC differentiation models and remyelination models to the promotion of remyelination, and verified the restorative effect of OGA inhibitors on the behavior of demyelination mouse models in vivo, thereby finding that it has huge potential application value in promoting remyelination in the treatment of demyelinating diseases. Completed the present invention on this basis.

[0093] OGA inhibitors of the present invention

[0094] 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 of the present invention that can promote O-linked beta-N-acetylglucosaminylation (O-GlcNAcylation or O-GlcNAc).

[0095] 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, J-01, TP-05.

[0096] 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.

[0097] The present invention also includes pharmaceutically acceptable salts of the compounds. The term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention with an acid. Suitable acids for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as 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, phenylmethanesulfonic acid, and benzenesulfonic acid; and acidic amino acids such as aspartic acid and glutamic acid.

[0098] The compounds of the present invention can be prepared by methods well known to those skilled in the art in the art, and there is no particular limitation on the reaction parameters of each step.

[0099] As used herein, in the compounds shown in the table above, if a chiral carbon atom exists, the chiral carbon atom may be in the R configuration, the S configuration, or a mixture of the two.

[0100] Promoting effect of the OGA inhibitor of the present invention on myelin regeneration

[0101] The OGA inhibitor of the present invention can increase O-linked beta-N-acetylglucosaminylation (O-GlcNAcylation or O-GlcNAc). O-GlcNAcylation is a dynamic post-translational modification process catalyzed by a pair of enzymes with opposing effects, namely O-linked glycosyltransferase (OGT) and glycoside hydrolase (OGA). OGT transfers beta-N-acetylglucosamine (O-GlcNAc) to the hydroxyl groups 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 cellular response to various stress conditions. Among them, O-GlcNAcylation modification is highly enriched in the brain and is crucial for the normal development and function of neurons.

[0102] Oligodendrocyte precursor / progenitor cells (OPCs) are resident progenitor cells distributed throughout the central nervous system (CNS). Their primary function is to form myelin sheaths that wrap around neuronal axons by proliferating and differentiating into oligodendrocytes (OLs), enabling rapid conduction of neural signals. Furthermore, OPCs can also maintain neuronal activity and synchronize neuronal circuits by differentiating into OLs, thereby affecting brain function. A variety of factors, including aging, genetic mutations, nutrition, autoimmune diseases, and / or injury, can lead to the destruction of myelin sheath structure and ultimately cause demyelinating diseases. Although functional impairments caused by myelin damage can be partially restored by remyelination, the efficiency of this myelin regeneration is very low. Clinical studies have shown that OPCs are present in large numbers in demyelinated areas but are unable to differentiate into mature OLs. Therefore, the failure of OPC differentiation is one of the factors that hinder myelin repair.

[0103] Through research, the inventors discovered that the OGA inhibitors of the present invention can increase the level of O-GlcNAcylation modification in OPCs, promote OPC differentiation, and further promote myelin production, thereby achieving remyelination after injury. In a demyelination model, the OGA inhibitors of the present invention significantly enhance myelin regeneration, reverse disease severity, and restore related body functions. They can be used to prevent or treat demyelination and related diseases.

[0104] Pharmaceutical compositions and administration methods of the present invention

[0105] Since the OGA inhibitors of the present invention have excellent effects in promoting myelination and / or regeneration, the OGA inhibitors of the present invention (including compounds and various crystal forms thereof, and pharmaceutically acceptable salts) and pharmaceutical compositions containing the OGA inhibitors of the present invention as the main active ingredient can be used to prevent and / or treat (stabilize, alleviate or cure) demyelination-related diseases.

[0106] 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. "Safe and effective amount" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, a pharmaceutical composition (e.g., one dose) contains 2 mg to 1000 mg of the OGA inhibitor of the present invention, more preferably, 10 to 500 mg of the OGA inhibitor of the present invention. Preferably, the "one dose" is a capsule, tablet, or injection. Preferably, the unit dosage form is a dose corresponding to the daily dose.

[0107] "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, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0108] There is no particular limitation on the administration route of the OGA inhibitor or pharmaceutical composition of the present invention. Representative administration routes include (but are not limited to): oral administration, parenteral administration (intravenous administration, intramuscular administration, or subcutaneous administration).

[0109] 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 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.

[0110] Solid dosage forms such as tablets, dragees, 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 opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0111] 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, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0112] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0113] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0114] Compositions for parenteral injection may comprise 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.

[0115] The OGA inhibitors of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0116] 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.

[0117] When using a pharmaceutical composition, a safe and effective amount of the targeting enhancer of the present invention is applied to a mammal (e.g., a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage. For a 60 kg human, the daily dosage is generally 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 patient's health condition, all of which are within the skill of a skilled physician.

[0118] Treatment

[0119] 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.

[0120] Preferably, the subject includes humans and non-human mammals (rodents, rabbits, monkeys, livestock, dogs, cats, etc.).

[0121] The main advantages of the present invention include:

[0122] (1) The present invention discovered for the first time 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.

[0123] (2) The present invention provides a group of OGA inhibitors that have excellent effects in promoting OPC differentiation and promoting myelin regeneration, and can be used to prevent and / or treat demyelination and its related diseases, or to prepare drugs for preventing and / or treating demyelination and its related diseases.

[0124] (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.

[0125] 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, for which specific conditions are not specified, are generally 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 according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0126] Equipment and materials

[0127] Table 2 Some main experimental materials

[0128] Example 1: Promoting effect of OGA inhibitors on OPC differentiation

[0129] 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 a rate of 2.5×10 4 pieces / cm 2The cells were seeded in Matrigel-coated 24-well plates and cultured 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. The culture medium was then replaced with OPC medium, and the small molecule compounds of the present invention were added to the OPC medium according to a concentration gradient. This process lasted for 3 days. OPCs were then induced to differentiate into OLs 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.). Different concentrations of the small molecule compounds of the present invention were continuously added to the OL medium for 3 days. The drug solvent DMSO (volume ratio v / V = 0.1%) was used as a control group to construct a directed 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.

[0130] The results are shown in Figures 1-9 and Tables 3 and 4. Tables 3 and 4 show the O4 + (Table 3), Mbp + (Table 4) Statistical results of cell percentage.

[0131] In Figures 1-8 B, C, Figure 9, and Tables 3 and 4, the statistical results show that compared with the control group, the O4 + , Mbp + The percentage of cells was significantly decreased, and the change in cell activity at this concentration was within 10% compared with the control group (Figure 1D-Figure 8D), and no obvious cell death occurred.

[0132] Tables 3 and 4 show that different small molecule compounds induce OPC 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 was observed. Based on the above criteria, the effective concentration range of each small molecule was determined and is shown in Tables 3 and 4.

[0133] Table 3 O4 under the action of different compounds + Cell percentage statistics

[0134] Note: *** indicates that when the concentration is 0.1nM, O4 + Cell percentage ≥ 20%; ** indicates that when the concentration is 0.1nM, O4 + The percentage of cells was less than 20%, when the concentration was 0.5nM, O4 + Cell percentage ≥20%; * indicates other.

[0135] Table 4 Mbp under the action of different compounds + Cell percentage statistics

[0136] Note: *** indicates that when the concentration is 0.1nM, Mbp + The percentage of cells is ≥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 other.

[0137] The above results show that the small molecule compound 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, and L5-06, can significantly increase O4 at very low concentrations (e.g., 0.1-5 nM). + The percentage of cells was significantly increased, 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 significantly increased Mbp at very low concentrations (e.g., 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.

[0138] Example 2: OGA inhibitors promote myelin regeneration in a demyelinated mouse model

[0139] To investigate whether OGA inhibitors could promote myelin regeneration in vivo, a toxin-induced model was used in which local injection of lysolecithin (lysophosphatidylcholine, LPC) induced demyelination in the corpus callosum (CC) of mice.

[0140] Using a toxin-induced model, mice were treated with a targeted injection of lysophosphatidylcholine (LPC) into the corpus callosum to induce acute demyelinating symptoms. Demyelination symptoms developed four days after injection, and remyelination began 14 days after injection. Therefore, the therapeutic effect of an OGA inhibitor on remyelination in the LPC model was evaluated between days 4 and 14 after injection. This model is recognized in the field as an animal model for evaluating remyelination. Mice were intraperitoneally administered daily from days 4 to 10 after LPC injection. A WT + saline group served as a blank control, and an LPC + saline group served as a negative control. On day 14, remyelination in the damaged area of ​​the corpus callosum was assessed by measuring the size of myelin sheaths labeled by FluoroMyelin and the area of ​​demyelinated areas marked by inflammatory regions with high Iba1 expression (Figures 1-8, Table 5). Each group consisted of two female mice and two male mice. The two-tailed T test was used for statistical analysis, and significance was defined as *P<0.05, **P<0.01, ***P<0.001.

[0141] FluoroMyelin can rapidly and selectively label myelin in brain slices, and its fluorescence intensity can be used as an indicator of myelin regeneration. Therefore, FluoroMyelin can be used to indicate the effect of OGA inhibitors on myelin regeneration in demyelinated areas.

[0142] Staining revealed that the saline-injected control group in the LPC mouse model exhibited severe demyelination. Quantitative analysis of the area of ​​demyelination in the corpus callosum lesion and the average level of demyelination after FluoroMyelin staining was performed. The results are shown in Figures 1-8 and Table 5.

[0143] Figures 1-8 (EF) show quantitative analysis of the myelin lesion area in the LPC + saline group and in the experimental group treated with small molecules. Drug treatment reduced the area of ​​myelin lesions. Fluoromyelin staining for myelin revealed a significant increase in the mean fluorescence intensity of corpus callosum lesions in the drug-treated group compared with the control group. Summary data are shown in Table 5.

[0144] Figures 1-8, F and G show the quantitative statistical results of the myelin damage area (Figure 1F) and the mean fluorescence intensity (MFI) of FluoroMyelin (Figure 1G), respectively.

[0145] Table 5 Myelin regeneration in mouse LPC model

[0146] Based on the results in Figures 1-8 and Table 5, it can be concluded 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.

[0147] Example 3: Effects of OGA inhibitors on behavioral recovery in a demyelination mouse model

[0148] 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.

[0149] In the accelerated rotating rod experiment, the initial speed was 4 rpm / min and the acceleration was 20 rpm / min. 2 The final speed was 40 rpm / min, and the maximum statistical time was 5 minutes. In the constant-speed rotarod test, the speed was 20 rpm / min, and the maximum statistical time was 1 minute. The effect of OGA inhibitors on the motor ability of demyelinated mice was evaluated by recording the time mice maintained movement on the accelerating and constant-speed rotarods.

[0150] In the accelerated study, 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 daily 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) from day 4 to 10 after model establishment. Behavioral scores were assessed on day 14. Each group consisted of two female mice and two 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.

[0151] The results showed that after drug treatment, the time the mice stayed on the rod was significantly increased. The data are shown in Table 6 and H of Figure 1-8.

[0152] 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 daily 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) from days 4 to 10 after model establishment. Behavioral scores were assessed on day 14. Each group consisted of 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.

[0153] The results showed that the time mice spent on the rod was significantly increased after treatment 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.

[0154] Table 6 The time mice remained on the rod

[0155] The above results show that the compound of the present invention can significantly improve the impaired motor ability of demyelination model mice.

[0156] discuss

[0157] O-linked β-N-acetylglucosamine glycosylation (O-GlcNAcylation, O-GlcNAc) is a post-translational modification (glycosylation) of proteins that is highly enriched in brain tissue and its levels decrease with aging. O-GlcNAcylation is catalyzed by glycosyltransferases (O-GlcNAc transferase, Ogt, responsible for glycosylation) and glycoside hydrolases (O-GlcNAcase, Oga, responsible for deglycosylation). Glycosylation can regulate various biological processes, including cell proliferation, migration, and differentiation, by modulating protein phosphorylation, protein stability, protein localization, protein-protein interactions, and gene expression. Studies have shown that glycosylation is involved in the maintenance of nervous system function and homeostasis. For example, decreased levels of O-GlcNAcylation in neural stem cells (NSCs) can affect cell fate determination in the hippocampus and impair cognitive function. However, whether O-GlcNAcylation plays a role in (re)myelination in the central nervous system has not been reported.

[0158] Currently, the main chemical drugs that promote O-GlcNAcylation modification are inhibitors of glycoside hydrolases (OGA). In the field of tumor research, abnormal O-GlcNAcylation modification in cells is directly correlated with the growth and proliferation, immune escape, and tumor metastasis of various tumor cells, such as leukemia, cervical cancer, and colorectal cancer. OGA inhibitors have begun to be used clinically to affect the proliferation of tumor cells by promoting O-GlcNAcylation modification in cells, thereby inhibiting tumor growth. However, the application of OGA inhibitors in the treatment of demyelination-related diseases has not been studied.

[0159] The root cause of myelination disorders is the inability of OPCs to 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 discovered 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 inhibitors of the present invention is to promote OPC differentiation ability by increasing the O-GlcNAcylation modification level of OPCs, thereby promoting the oligodendrocyte lineage development process 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.

[0160] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

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) Prevention and / or treatment of demyelination and its related diseases; (b) promoting the generation, maintenance and / or proliferation of oligodendrocyte precursor / progenitor 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 The OGA inhibitor includes a compound selected from the following group or a pharmaceutically acceptable salt thereof: (3) Compound represented by formula III: in, X5 is -N- or -CH-; X6 is -N=, -O- or -S-; W3 is H, F, -R2- or -OR 2 , R 2 is zero, substituted or unsubstituted C1-C8 alkyl; Ring C is a substituted or unsubstituted 6-10 membered aryl group, 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; (4) Compound represented by formula IV:

4. The use according to claim 2, 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-.

5. The use according to claim 2, 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.

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

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

8. The use according to claim 1, characterized in that The demyelination and its related diseases are selected from the following groups: multiple sclerosis, neuromyelitis optica, spinal cord injury, demyelination caused by brain injury, demyelination, psychiatric diseases caused by demyelination, degenerative diseases and / or neurological dysfunction, myelin regeneration disorders caused by aging, or a combination thereof.

9. 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.

10. 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.

11. 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.

12. The pharmaceutical composition according to claim 11, characterized in that The OGA inhibitor is selected from the following group: a compound of formula III or a pharmaceutically acceptable salt thereof; a compound of formula IV (TP-05), or a combination thereof.

13. The pharmaceutical composition according to claim 11 or 12, characterized in that The OGA inhibitor is 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, A1-01, AT1-01, AT1-02, B1-01, B1-02, B2-1-01, B2-1-02, B 2-2-01, B2-2-02, B5-01, B5-02, A2-01, A2-02, A3-01, A3-02, A5-01, A5-02, J-01, TP-05.

14. 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.

15. The kit according to claim 14, wherein: The OGA inhibitor is selected from the following group: a compound of formula III or a pharmaceutically acceptable salt thereof; a compound of formula IV (TP-05), or a combination thereof.

16. A method for treating a demyelination-related disease, characterized in that: The method comprises the steps of administering a pharmaceutical composition containing a therapeutically effective amount of an OGA inhibitor to a subject in need of treatment.

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