COMPOSITION, COMPRISING THIAMINE DERIVATIVE, FOR PREVENTION OR TREATMENT OF HYPERCORTISOLEMIA

Abstract

The present invention relates to a composition for prevention or treatment of hypercortisolemia, the composition comprising allithiamine, fursultiamine, and benfotiamine, which are all derivatives of thiamine, or a mixed composition thereof. The composition for prevention or treatment of hypercortisolemia of the present invention can effectively treat or alleviate symptoms or disorders resulting from excessive secretion of cortisol, without surgical operations, and does not exhibit side effects caused by conventional medications.

Claims

1-6. (canceled)

7. A method for preventing or treating of hypercortisolism, the method comprising: a step of administering to the subject a pharmaceutical composition comprising at least one compound selected from the group consisting of allithiamine, fursultiamine, benfotiamine, and pharmaceutically acceptable salts thereof.

8. The method of claim 7, wherein the hypercortisolism is selected from the group consisting of Cushing's syndrome, Cushing's disease, and ectopic adrenocorticotropic hormone syndrome.

9. A method for preventing or treating of cortisol-induced hypertension, diabetes, obesity, depression or acute stress, the method comprising: a step of administering to the subject a pharmaceutical composition comprising at least one compound selected from the group consisting of allithiamine, fursultiamine, benfotiamine, and pharmaceutically acceptable salts thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0051] FIG. 1A is a graph depicting the test results of cortisol synthesis inhibitory ability after the treatment of the human adrenocortical carcinoma cell line NCI-H295R with allithiamine.

[0052] FIG. 1B is a graph depicting the test results of cortisol synthesis inhibitory ability after the treatment of the human adrenocortical carcinoma cell line NCI-H295R with fursultiamine.

[0053] FIG. 1C is a graph depicting the test results of cortisol synthesis inhibitory ability after the treatment of the human adrenocortical carcinoma cell line NCI-H295R with benfotiamine.

[0054] FIG. 2A is a graph depicting the body weight checking results to investigate the cortisol-induced obesity alleviating effect of fursultiamine in Cushing's syndrome animal models.

[0055] FIG. 2B shows images and a graph, which depict the fat mass checking results, to investigate the cortisol-induced obesity alleviating effect of fursultiamine in Cushing's syndrome animal models.

[0056] FIG. 3 shows a graph depicting the results of verifying cortisol inhibitory effects of fursultiamine in Cushing's syndrome animal models.

[0057] FIG. 4 shows a graph depicting the results of verifying cortisol synthesis inhibitory ability of fursultiamine in the stress-induced test.

BEST MODE FOR CARRYING OUT THE INVENTION

[0058] The present disclosure is directed to a pharmaceutical composition for prevention or treatment of hypercortisolism, the pharmaceutical composition containing at least one compound selected from the group consisting of thiamine derivatives and pharmaceutically acceptable salts thereof, the thiamine derivatives being allithiamine, fursultiamine, and benfotiamine.

Mode for Carrying Out the Invention

[0059] Hereinafter, the present disclosure will be described in more detail with reference to examples. These examples are provided only for the purpose of illustrating the present disclosure in more detail, and therefore, according to the purpose of the present disclosure, it would be apparent to a person skilled in the art that these examples are not construed to limit the scope of the present disclosure.

EXAMPLE 1

Verification of Cortisol Reduction effects by thiamine derivatives in vitro

[0060] Cortisol secretion inhibitory effects by allithiamine (Shanghai iChemical Technology CO., LTD., China), fursultiamine (Toronto Research Chemicals, Canada), or benfotiamine (Sigma-Aldrich, St. Louis, Mo., USA), which are thiamine (vitamin B1) based compounds, were tested. For this, cortisol reduction effects in adrenal tumors that excessively secret cortisol were measured.

[0061] NCI-H295R (ATCC #CRL-2128) cells, a human adrenocortical carcinoma cell line, were dispensed into 24-well plates at 2.5×10.sup.5 cells/well. Then, the cells were incubated in DMEM/F12 growth media containing 2.5% Nu-serum, 1% ITS (insulin, transferrin, and selenium), bovine serum albumin (BSA), linoleic acid, and penicillin/streptomycin for 48 hours.

[0062] Thereafter, the media were exchanged with serum-free media, and then the NCI-H295R cells were pretreated with allithiamine, fursultiamine, or benfotiamine at concentrations of 20 μM, 50 μM, and 100 μM for 12 hours. The cells were treated with 8-bromoadenosine 3′, 5′-cyclic monophosphate (8-Br-cAMP) at a concentration of 0.5 mM to increase cortisol synthesis, and again treated with allithiamine, fursultiamine, or benfotiamine at 20 μM, 50 μM, and 100 μM, followed by reaction for 24 hours. After the reaction, cell cultures were collected, and the amount of cortisol produced in the cells was measured using an enzyme-linked immunosorbent assay (ELISA) kit (Enzo Life Sciences, Inc., N.Y., USA).

[0063] It was verified that all of allithiamine, fursultiamine, and benfotiamine significantly inhibited the cortisol production, increased by the stimulation of 0.5 mM 8-Br-cAMP, in a dose-dependent manner.

[0064] Compared with the allithiamine non-treatment group (with 8-Br-cAMP treatment), the allithiamine treatment groups showed a reduction in cortisol production of 50.7% at 50 μM and 78.7% at 100 μM (see Table 1 and FIG. 1A). The fursultiamine treatment groups showed a reduction in cortisol production of 43% at 50 μM and 52.4% at 100 μM (see Table 2 and FIG. 1B). The benfotiamine treatment groups showed a reduction in cortisol production of 22.6% at 50 μM and 44.3% at 100 μM (see Table 3 and FIG. 1C) (**p<0.01, ***p<0.001 vs. 8-Br-cAMP treatment group).

TABLE-US-00001 TABLE 1 Cortisol production inhibitory effect of allithiamine 8-Br-cAMP (mM) 0 0.5 0.5 0.5 0.5 Allithiamine (μM) 0 0 20 50 100 Cortisol production concentration 3.5 13.0 10.5 6.4 2.8 (ng/mL)

TABLE-US-00002 TABLE 2 Cortisol production inhibitory effect of fursultiamine 8-Br-cAMP (mM) 0 0.5 0.5 0.5 0.5 Fursultiamine (μM) 0 0 20 50 100 Cortisol production concentration 4.5 15.7 14.1 9.0 7.5 (ng/mL)

TABLE-US-00003 TABLE 3 Cortisol production inhibitory effect of fursultiamine 8-Br-cAMP (mM) 0 0.5 0.5 0.5 0.5 Benfotiamine (μM) 0 0 20 50 100 Cortisol production concentration 4.1 15.5 14.0 12.0 8.6 (ng/mL)

EXAMPLE 2

Verification of Cortisol Reduction Effects by Thiamine Derivatives in vivo

[0065] 2-1. Tests using Cushing's Syndrome Animal Models

[0066] To investigate high-cortisol inhibitory effects, C57BL/6J normal mice (WT-con) aged 8 to 9 weeks and corticotropin-releasing hormone (CRH)-overexpressed mice, which are Cushing's syndrome animal models, were used. One group of CRH-overexpressed mice were orally administered with 60 mg/kg fursultiamine once a day for 9 weeks (CRH tg-Fur 60 mpk).

[0067] During this period, the body weight change was monitored by measuring body weights once a week. To check the concentration of corticosterone (in cases of rodents), blood was collected from the tail vein and serum was isolated. The concentration of corticosterone was measured from the isolated serum by using an enzyme-linked immunosorbent assay (ELISA) kit (Enzo Life Sciences, Inc., N.Y., USA). The fat mass was measured 6 weeks after administration of the drug by using a body composition analyzer (MiniSpec LF 50, Bruker Optics, Germany), and the fat distribution was again monitored with micro CT images 8 weeks after administration.

[0068] As can be verified in FIG. 2A and Table 4, the CRH-overexpressed mice showed a greater increase in body weight compared with the normal mice, whereas the CRH-overexpressed mice administered with fursultiamine showed a smaller increase in body weight compared with the control (CRH-overexpressed mice).

TABLE-US-00004 TABLE 4 CRH- CRH-overexpressed mice Normal overexpressed administered with Weeks mice mice fursultiamine 0 24.0 19.0 18.4 1 24.9 18.5 17.8 2 24.9 18.2 17.8 3 24.9 19.4 18.2 4 24.9 20.0 18.1 5 24.9 20.5 18.4 6 25.0 20.4 17.9 7 25.8 22.8 19.8 8 26.5 22.4 19.7 9 26.4 21.9 19.2

[0069] As can also be verified in FIG. 2B and Table 5, the fat mass of the CRH-overexpressed mice was greater compared with the normal mice, whereas the fat mass of the CRH-overexpressed mice administered with fursultiamine was smaller compared with the control.

TABLE-US-00005 TABLE 5 CRH- CRH-overexpressed mice Normal overexpressed administered with mice mice fursultiamine Body 16.9 28.0 23.4 weight (%)

[0070] As can be verified in FIG. 3 and Table 6, the corticosterone synthesis concentration of the CRH-overexpressed mice was greater compared with the normal mice, whereas the corticosterone synthesis concentration of the CRH-overexpressed mice administered with fursultiamine was smaller compared with the normal group.

TABLE-US-00006 TABLE 6 CRH- CRH-overexpressed mice Normal overexpressed administered with mice mice fursultiamine Corticosterone 37.0 130.5 87.1 (ng/mL)

[0071] Stress-Induced Test

[0072] For the restraint stress test, 8-week-old male mice were used, and were orally administered with fursultiamine once 24 hours before and once 2 hours before stress induction. For induction of stress, the mice were individually placed head-first into well-ventilated 50 mL polypropylene tubes, which were then tied with a cap of the 50 mL tube, and the mice were confined for 2 hours. Before stress induction, 1 hour and 2 hours after induction, and at 1 hour of recovery, blood was collected from the tail vein and serum was isolated, and then the concentration of corticosterone was measured using ELISA kit.

[0073] As can be verified in FIG. 4 and Table 7, the corticosterone levels increased after stress induction, and the rate of increase in corticosterone level decreased in the fursultiamine administration group.

TABLE-US-00007 TABLE 7 Fursultiamine Time (min) Control administration group 0 76.6 63.6 60 300.0 260.4 120 255.1 261.4 180 188.3 105.8

Industrial Applicability

[0074] The present disclosure relates to a composition containing a thiamine derivative for prevention or treatment of hypercortisolism and, more specifically, to a composition for prevention or treatment of hypercortisolism, the composition containing a thiamine derivative, a salt thereof, or a mixture composition thereof, the thiamine derivative being allithiamine, fursultiamine, or benfotiamine.