PHARMACEUTICAL COMPOSITION FOR PREVENTING OR TREATING GRAFT VERSUS HOST DISEASE COMPRISING THIAMINE DERIVATIVES

Abstract

The present invention relates to a pharmaceutical composition for preventing, treating, or ameliorating graft versus host disease (GVHD), comprising thiamine derivatives, in which the active ingredient of the pharmaceutical composition is fursultiamine, which can alleviate weight loss found in patients with GVHD, reduce the infiltration of liver and colon inflammatory cells, and effectively inhibit the production of allogeneic CD4+ IFNγ+ T cells, and thus can be effectively used for the prevention, treatment, or amelioration of GVHD.

Claims

1-8. (canceled)

9. A method for preventing or treating graft versus host disease (GVHD), comprising step of: administering a pharmaceutical composition comprising at least one compound selected from the group consisting of a thiamine derivative represented by chemical formula 1 below and a pharmaceutically acceptable salt thereof to a subject ##STR00013## wherein R is any one selected from the group consisting of ##STR00014##

10. The method of claim 9, wherein R is ##STR00015##

11. The method of claim 9, wherein the thiamine derivative is at least one selected from the group consisting of fursultiamine, allithiamine, and benfotiamine.

12. The method of claim 9, wherein the graft versus host disease occurs in cancer patients undergoing hematopoietic stem cell transplantation after receiving irradiation.

13. A method for alleviating graft versus host disease (GVHD), comprising step of: administering a food composition comprising at least one compound selected from the group consisting of a thiamine derivative represented by chemical formula 1 below and a pharmaceutically acceptable salt thereof to a subject ##STR00016## wherein R is any one selected from the group consisting of ##STR00017##

14. The method of claim 13, wherein R is ##STR00018##

15. The method of claim 13, wherein the thiamine derivative is at least one selected from the group consisting of fursultiamine, allithiamine, and benfotiamine.

16. The method of claim 13, wherein the graft versus host disease occurs in cancer patients undergoing hematopoietic stem cell transplantation after receiving irradiation.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0042] FIG. 1 is a graph illustrating an CD4+IFNg+ T cell generation inhibiting effect of a thiamine derivative in the mixed lymphocyte reaction induction experiment using mouse cells according to an example of the present disclosure.

[0043] FIG. 2A is a schematic diagram of the construction of graft versus host disease (GVHD) model mice and the administration procedure of fursultiamine according to an example of the present disclosure.

[0044] FIG. 2B is a graph illustrating weight changes of the graft versus host disease model mice according to an example of the present disclosure.

[0045] FIG. 2C is a graph illustrating the change in amount of generation of CD4+ T cells in the spleens isolated from the graft versus host disease model mice according to an example of the present disclosure.

[0046] FIG. 2D is a graph illustrating the change in amount of generation of CD4+IFNg+ T cells in the spleens isolated from the graft versus host disease model mice according to an example of the present disclosure.

[0047] FIG. 2E shows hematoxylin and eosin (H&E) staining images of liver and large intestine tissues isolated from the graft versus host disease model mice according to an example of the present disclosure.

BEST MODE FOR CARRYING OUT THE INVENTION

[0048] The present disclosure is directed to a pharmaceutical composition for prevention or treatment of graft versus host disease (GVHD), the pharmaceutical composition containing at least one compound selected from the group consisting of a thiamine derivative represented by chemical formula 1 below and a pharmaceutically acceptable salt thereof:

##STR00011##

[0049] wherein R is any one selected from the group consisting of

##STR00012##

MODE FOR CARRYING OUT THE INVENTION

[0050] Hereinafter, the present disclosure will be described in more detail by the following examples. However, these exemplary embodiments are used only for illustration, and the scope of the present disclosure is not limited by these exemplary embodiments.

[0051] Throughout the present specification, the “%” used to express the concentration of a specific material, unless otherwise particularly stated, refers to (wt/wt) % for solid/solid, (wt/vol) % for solid/liquid, and (vol/vol) % for liquid/liquid.

Example 1

Effect of Thiamine Derivatives on Allo-Reactive CD4+IFNg+ T Cell Generation

[0052] A high amount of CD4+IFNg+ T cell generated can be inferred to allow the proliferation of T cells against allogenic antigens, thereby attacking the body organs of a patient. Therefore, a graft versus host disease (GVHD) in vitro validation model was subjected to a mixed lymphocyte reaction to compare the degree of allo-reactive CD4+IFNg+ T cell generation among thiamine derivatives by flow cytometry.

[0053] Specifically, BALB/c mice were sacrificed by cervical dislocation, and then the spleens were extracted, followed by erythrocyte lysis using ACK lysing buffer. Thereafter, the isolated spleen cells were irradiated at 2500 cGy.

[0054] A syngeneic control group was prepared by mixing 8×105 BALB/c mouse-derived irradiated splenocytes and 8×105 BALB/c-derived irradiated CD4+ T cells, and a positive control group was prepared by mixing 8×105 BALB/c mouse-derived irradiated splenocytes and 8×105 C57BL6/J mouse-derived CD4+ T cells. The positive group was treated with 10, 50, and 100 uM benfotiamine, 1, 5, and 10 uM allithiamine, and 10, 50, and 100 uM fursultiamine to prepare experimental groups.

[0055] Thereafter, the cells were cultured in RPMI1640 (10% FBS, 1% penicillin streptomycin) at 37° C. for 5 days, and the cells were separated, followed by flow cytometry, to investigate the degree of CD4+IFNg+ T cell generation.

TABLE-US-00001 TABLE 1 Syngeneic Positive Benfotiamine (μM) Allithiamine (μM) Fursultiamine (μM) control control 10 50 100 1 5 10 10 50 100 Amount of 15.3 37.4 36.8 43.9 40.1 35.2 25.7 23.9 34.9 13.6 12.5 generation (%)

[0056] As can be seen from Table 1 and FIG. 1, the treatment with benfotiamine showed no significant difference in the degree of generation of CD4+IFNg+ T cells, compared with the positive control, regardless of the concentration of benfotiamine. However, allithiamine at 5 uM or more and fursultiamine at 50 uM or more showed a significant reduction compared with the positive control, in a concentration dependent manner.

Example 2

Treatment Effect of Fursultiamine in GVHD Disease Model

[0057] Referring to FIG. 2A, 5-week-old BALB/c male mice were acclimatized for 1 week, irradiated with 800 cGy radiation throughout the body, and then injected with 5×106 C57BL/6J-derived bone marrow cells and 1×107 splenocytes derived therefrom via IV injection, thereby constructing a GVHD in vivo model (positive control group, Positive). The syngeneic control group (Syngeneic) was constructed in the same manner as above while BALB/c-derived bone marrow cells were selected as the bone marrow cells to be injected.

[0058] The fursultiamine administration group (Fur50) was constructed by daily oral gavage of fursultiamine HCl at 50 mg/kg to the GVHD in vivo model.

[0059] The treatment effect of fursultiamine was investigated on the constructed experimental groups by measuring the change in weight every 3 days.

[0060] As can be seen from FIG. 2B, the fursultiamine administration group showed a significant recovery compared with the GVHD in vivo model on Day 12 of oral administration.

[0061] On Day 14, the comparison between the amounts of generation of CD4+ T cells and CD4+IFNg+ T cells, which are major factors causing GVHD, was performed in the spleens isolated from the mice.

TABLE-US-00002 TABLE 2 Syngeneic Positive Fursultiamine control control administration Amount of generation of 6.9 21.1 13.4 CD4+ T cells (%) Amount of generation of 3.2 18.3 12.6 CD4+IFNg+ T cells (%)

[0062] As can be seen from Table 2 and FIGS. 2C and 2D, the amounts of generation of allogenic CD4+ T cells and CD4+IFNg+ T cells were reduced by fursultiamine administration. On Day 14 of inducing the GVHD in vivo model, liver and large intestine tissues were extracted from the mice, fixed in 4% paraformaldehyde for 48 hours, and then embedded in paraffin. The resultant tissues were sliced into 5 μm sections and then subjected to hematoxylin and eosin (H&E) staining. (bar=100 um)

[0063] As can be seen from FIG. 2E, the infiltration of immune cells (indicated by the red arrow) was observed in the liver tissue of the positive control group, but the infiltration of immune cells was reduced in the fursultiamine administration group.

INDUSTRIAL APPLICABILITY

[0064] The present disclosure relates to a composition containing a thiamine derivative for prevention, treatment, or alleviation of graft versus host disease and, more specifically, to a composition containing fursultiamine for prevention, treatment, or alleviation of graft versus host disease.