PHARMACOLOGICAL INTERVENTION FOR LIVER REGENERATION THROUGH GSK-3? INHIBITION AND RELATED METHODS

Abstract

ABSTRACT The invention relates to a composition for promoting liver regeneration and providing a hepatoprotective effect. It comprises a selective GSK-3ß inhibitor, which is a chemical compound that inhibits GSK-3ß activity, and a pharmaceutically acceptable carrier. The selective GSK-3ß inhibitor accumulates ß-catenin and activates the Wnt/ß-catenin signaling pathway in the liver, thereby stimulating liver regeneration and repair. The composition can be formulated for oral, intravenous, or intraperitoneal administration. Additionally, a solid oral composition is described, which includes the selective GSK-3ß inhibitor and one or more pharmaceutically acceptable excipients, formulated as a tablet with specific amounts of diluents, binders, disintegrants, and lubricants. A method for promoting liver regeneration and providing a hepatoprotective effect by administering the composition to a subject with liver toxicity or damage is also disclosed. Reference Fig 1

Specification

Description:TECHNICAL FIELD
[0001] The present disclosure relates generally to pharmaceutical compositions for liver disease treatment, specifically oral formulations of CHIR99021 and their effects on liver function.
BACKGROUND
[0002] Liver diseases and injuries, such as those induced by toxins like cyclophosphamide, pose significant health challenges due to the liver's critical role in metabolism, detoxification, and overall homeostasis. The liver's ability to regenerate is a unique and vital characteristic, yet in cases of severe damage, this regenerative capacity can be overwhelmed, leading to chronic liver conditions or failure. Current therapeutic strategies often focus on managing symptoms and preventing further damage, but there is a pressing need for interventions that can actively promote liver regeneration and repair.

[0003] The Wnt/ß-catenin signaling pathway is a crucial regulator of liver development and regeneration. It plays a significant role in controlling cell proliferation, differentiation, and survival. Dysregulation of this pathway has been implicated in various liver pathologies, including cancer and fibrosis. Therefore, modulating this pathway presents a promising avenue for therapeutic intervention. However, the challenge lies in effectively targeting the pathway to enhance liver regeneration without triggering adverse effects. This necessitates a deeper understanding of the molecular mechanisms involved and the development of targeted pharmacological agents that can safely and effectively stimulate liver repair processes.
SUMMARY
[0004] In accordance with embodiments, a composition is provided for promoting liver regeneration and providing a hepatoprotective effect. The composition comprises a selective GSK-3ß inhibitor, which is a chemical compound that inhibits GSK-3ß activity, and a pharmaceutically acceptable carrier. The selective GSK-3ß inhibitor functions by accumulating ß-catenin and activating the Wnt/ß-catenin signaling pathway in the liver, thereby stimulating liver regeneration and repair.
[0005] In accordance with other embodiments, the composition includes the chemical compound CHIR99021 as the selective GSK-3ß inhibitor.
[0006] In further embodiments, the selective GSK-3ß inhibitor is present in an amount effective to enhance cellular growth and repair in the liver.
[0007] In yet other embodiments, the selective GSK-3ß inhibitor is present in an amount effective to provide organ protection in the liver.
[0008] In additional embodiments, the composition is formulated for various methods of administration, including oral, intravenous, or intraperitoneal administration.
[0009] In accordance with other embodiments, a solid oral composition is provided for promoting liver regeneration and providing a hepatoprotective effect. This composition comprises a selective GSK-3ß inhibitor and one or more pharmaceutically acceptable excipients. The selective GSK-3ß inhibitor accumulates ß-catenin and activates the Wnt/ß-catenin signaling pathway in the liver, thereby stimulating liver regeneration and repair.
[0010] In further embodiments, the solid oral composition includes the chemical compound CHIR99021 as the selective GSK-3ß inhibitor.
[0011] In yet other embodiments, the selective GSK-3ß inhibitor in the solid oral composition is present in an amount effective to enhance cellular growth and repair in the liver.
[0012] In additional embodiments, the solid oral composition is formulated as a tablet comprising about 1 mg to about 1000 mg of the selective GSK-3ß inhibitor, about 50 mg to about 500 mg of a diluent, about 5 mg to about 100 mg of a binder, about 1 mg to about 50 mg of a disintegrant, and about 0.1 mg to about 10 mg of a lubricant.
[0013] In accordance with yet other embodiments, a method is provided for promoting liver regeneration and providing a hepatoprotective effect. The method involves administering a composition comprising a selective GSK-3ß inhibitor to a subject with liver toxicity or damage. The selective GSK-3ß inhibitor accumulates ß-catenin and activates the Wnt/ß-catenin signaling pathway in the liver of the subject, thereby stimulating liver regeneration and repair.
BRIEF DESCRIPTION OF THE DRAWINGS
[0014] Figure1 illustrates administering a GSK-3ß inhibitor to stimulate liver regeneration and repair.
[0015] Figure 2 is Diagrammatic illustration of the protocol of cyclophosphamide-induced hepatotoxicity.
[0016] Figure 3 illustrate Effect of GSK-3 ß inhibitor (CHIR99021) on body weight in cyclophosphamide-induced hepatotoxicity rodents.
[0017] Figure 4 illustrate TBARS Levels for Oxidative Stress
[0018] Figure 5, 6, 7, and 8 illustrate Liver Enzyme Profile
[0019] Figures 9, 10, and 11 illustrate Effect of GSK-3ß Inhibitor (CHIR99021) on Total Protein, Albumin, and Globulin
[0020] Figure 12 illustrate IL-6 Levels and Inflammatory Response
[0021] Figure 13 illustrate effect of GSK-3 ß inhibitor (CHIR99021) in mean fold change of gene expression levels ß-catenin in cyclophosphamide induced hepatotoxicity rodents.
[0022] Figure 14 illustrate the effect of GSK-3 ß inhibitor (CHIR99021) on mitotic index in Cyclophosphamide induced hepatotoxicity rodents.
[0023] Figure 15 illustrate the effect of GSK-3 ß inhibitor (CHIR99021) on liver histological changes in cyclophosphamide induced hepatotoxicity rodents.

DETAILED DESCRIPTION
[0024] According to a potential configuration, the inhibition of GSK-3ß may be achieved through the administration of CHIR99021, which can lead to the accumulation of ß-catenin. This accumulation may activate the Wnt/ß-catenin signaling pathway, which can stimulate liver regeneration and repair. The process may involve an increase in liver cell proliferation, which can contribute to the hepatoprotective effect. The selective GSK-3ß inhibitor, CHIR99021, may be formulated with a pharmaceutically acceptable carrier to facilitate its administration. The composition may be designed to enhance cellular growth and repair in the liver, potentially providing organ protection. The formulation may be suitable for various routes of administration, including oral, intravenous, or intraperitoneal. The chemical compound CHIR99021 may be present in an amount effective to achieve the desired therapeutic outcomes. The potential for liver regeneration and protection may be further supported by the activation of the Wnt/ß-catenin signaling pathway, which may play a role in the process. The overall approach may focus on the stimulation of liver regeneration by targeting the GSK-3ß enzyme, thereby promoting a protective and regenerative effect on the liver.
[0025] FIG. 1 is a flowchart illustrating a method for promoting liver regeneration and providing a hepatoprotective effect through the administration of a selective GSK-3ß inhibitor to a subject with liver toxicity or damage.
[0026] Step 100 involves administering the selective GSK-3ß inhibitor, such as CHIR99021, to a subject experiencing liver toxicity or damage.
[0027] Step 102 includes the inhibition of GSK-3ß and the accumulation of ß-catenin, which activates the Wnt/ß-catenin signaling pathway.
[0028] Step 104 involves stimulating liver regeneration and repair processes.
[0029] The composition may be formulated for various routes of administration, including oral, intravenous, or intraperitoneal. The method may be beneficial in addressing liver damage induced by factors such as cyclophosphamide and improving the structure and function of hepatocytes in damaged liver tissues. The pharmacological intervention may also be evaluated through various models, including partial hepatectomy in rodents, to assess its impact on liver regeneration and repair.
[0030] In the context of the process, the selective GSK-3ß inhibitor can lead to the accumulation of ß-catenin and the activation of the Wnt/ß-catenin signaling pathway within the liver of the subject. This step is crucial in the mechanism by which liver regeneration and repair are potentially stimulated. The selective GSK-3ß inhibitor, identified as CHIR99021, may inhibit GSK-3ß activity, thereby allowing ß-catenin to accumulate. This accumulation may activate the Wnt/ß-catenin signaling pathway, which is a pathway in cellular processes such as proliferation and differentiation. The activation of this pathway may result in increased liver cell proliferation, which can contribute to liver regeneration. The process may also provide a hepatoprotective effect, potentially safeguarding the liver from further damage. The involvement of CHIR99021 in this process may be significant, as it may enhance the cellular growth and repair mechanisms within the liver, thereby promoting overall liver health. The potential for this intervention to stimulate liver regeneration and provide a hepatoprotective effect may be of particular interest in scenarios involving liver toxicity or damage.
[0031] The process may involve stimulating liver regeneration and repair. This step may be associated with liver regeneration and repair, which may be linked to the stimulation of liver regeneration and the provision of a hepatoprotective effect. The actions associated with this step may include the stimulation of liver regeneration and repair, which may be achieved through the inhibition of GSK-3ß, the accumulation of ß-catenin, and the activation of the Wnt/ß-catenin signaling pathway. These actions may lead to increased liver cell proliferation, which may contribute to the hepatoprotective effect. The selective GSK-3ß inhibitor, such as CHIR99021, may play a role in this process by inhibiting GSK-3ß activity, thereby allowing for the accumulation of ß-catenin and the subsequent activation of the Wnt/ß-catenin signaling pathway. This pathway may be essential for promoting liver regeneration and repair, as it may enhance cellular growth and repair mechanisms within the liver. The potential for liver regeneration and repair may be further supported by the presence of a pharmaceutically acceptable carrier, which may facilitate the delivery and effectiveness of the selective GSK-3ß inhibitor. Overall, the process described may highlight the potential for pharmacological intervention to stimulate liver regeneration and provide a hepatoprotective effect through the modulation of key signaling pathways and cellular processes.
[0032] According to an embodiment, the inhibition of GSK-3ß may be achieved through the administration of CHIR99021, which may lead to an increase in liver cell proliferation. The composition for promoting liver regeneration and providing a hepatoprotective effect comprises a selective GSK-3ß inhibitor, specifically CHIR99021, and a pharmaceutically acceptable carrier. The composition may be formulated for oral, intravenous, or intraperitoneal administration.
[0033] Oral Composition and Excipients:
[0034] In an embodiment, the present composition comprises CHIR99021, which is present in a range of 5 wt.% to 45 wt.% of the composition. The formulation is administered orally and is an immediate release oral solid dosage form.
[0035] The composition comprises one or more pharmaceutically acceptable excipients, present in a range of 65 wt % to 95 wt % of the composition. The excipients may be selected from those well-known in the art, including antiadherants, bulking agents, glidants, disintegrating agents, lubricants, colorants, and binding agents.
[0036] The bulking agents employed can be dicalcium phosphate, microcrystalline cellulose, calcium sulfate, and starches.
[0037] Disintegrating agents may include crospovidone, sodium starch glycolate, microcrystalline cellulose, starches, and croscarmellose sodium.
[0038] Antiadherants and glidants can include sodium lauryl sulfate, talc, corn starch, silicon dioxide, and metal stearates.
[0039] Lubricants can be sodium stearate, magnesium stearate, stearic acid, sodium stearyl fumarate, sterotex, talc, colloidal silica dioxide, waxes, calcium stearate, and the like.
[0040] Binding agents can include starch, methyl cellulose, hydroxypropyl methylcellulose, polyvinyl pyrollidone, carboxymethyl cellulose, and the like.
[0041] The composition is preferably in tablet form and can also be produced as tablets in the form of hard gelatin capsules or packaged into sachets. The tablets are prepared using methods known in the art, including a wet granulation method and a direct compression method, with a preference for the direct compression method.
[0042] The oral composition of CHIR99021 may include about 50 mg to about 500 mg of a diluent selected from the group consisting of lactose, microcrystalline cellulose, starch, and combinations thereof.
[0043] The oral composition of CHIR99021 may include about 5 mg to about 100 mg of a binder selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and combinations thereof.
[0044] The oral composition of CHIR99021 may include about 1 mg to about 50 mg of a disintegrant selected from the group consisting of croscarmellose sodium, crospovidone, sodium starch glycolate, and combinations thereof.
[0045] The oral composition of CHIR99021 may include about 0.1 mg to about 10 mg of a lubricant selected from the group consisting of magnesium stearate, stearic acid, talc, and combinations thereof.
[0046] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure.
EXAMPLES:
[0047] Experimental Details
1. Experimental Design and Animal Model as given in Figure 2
2. The study used 30 male Wistar albino rats divided into five groups (n=6). Each group received different treatments to evaluate the hepatoprotective effects of CHIR99021 against cyclophosphamide (CYP)-induced hepatotoxicity. Animals were housed under controlled conditions (21±1°C, 45-55% humidity, 12-h light/dark cycle) and provided standard chow and water ad libitum.

[0048] Results Summary
1. Body Weight Analysis as given in Figure 3
o No significant change in body weight was observed between groups on the 0th and 7th days. However, the CYP-treated group showed a significant reduction in body weight on the 14th day compared to the control and other treatment groups.
2. TBARS Levels for Oxidative Stress as given in Figure 4
o CYP treatment significantly increased TBARS levels, indicating lipid peroxidation. Both 6.25 mg/kg and 12.5 mg/kg doses of CHIR99021 significantly reduced TBARS levels, with the higher dose demonstrating superior antioxidant effects.
3. Liver Enzyme Profile as given in Figures 5, 6, 7, and 8
o SGPT, SGOT, ALP, and TB levels were elevated in the CYP-treated group, reflecting hepatotoxicity. Treatment with CHIR99021, especially at 12.5 mg/kg, significantly reduced these enzyme levels, surpassing the effectiveness of Silymarin (200 mg/kg) and the 6.25 mg/kg CHIR99021 dose.
4. Effect of GSK-3ß Inhibitor (CHIR99021) on Total Protein, Albumin, and Globulin is given in Figures 9, 10, and 11
o The CYP-treated group showed a significant decrease (P<0.05) in total protein, albumin, and globulin levels compared to the control group. Treatment with CHIR99021 (6.25 mg/kg and 12.5 mg/kg) significantly restored these levels, with the 12.5 mg/kg dose showing the most improvement, surpassing both Silymarin and the lower CHIR99021 dose.
5. IL-6 Levels and Inflammatory Response (Figure 12)
o The CYP group exhibited elevated IL-6 levels, indicating inflammation. CHIR99021 reduced IL-6 levels, with the 12.5 mg/kg dose showing stronger anti-inflammatory effects than Silymarin and the lower CHIR99021 dose.
6. ß-Catenin Gene Expression (Figure 13)
o CHIR99021 significantly increased ß-catenin expression, activating the Wnt/ß-catenin pathway responsible for liver regeneration. The 12.5 mg/kg dose demonstrated the highest fold change in gene expression compared to the CYP and control groups.
7. Mitotic Index Analysis (Figure 14)
o CHIR99021-treated animals showed higher mitotic index values, indicating increased hepatocyte proliferation. The 12.5 mg/kg group exhibited the highest mitotic activity, suggesting enhanced liver regeneration.
8. Histological Analysis of Liver Regeneration (Figure 15)
o Histopathological examination revealed significant liver regeneration in the CHIR99021-treated groups, with the 12.5 mg/kg dose showing the most prominent regenerative changes compared to the other groups.

[0049] These results confirm that CHIR99021 at a higher dose (12.5 mg/kg) significantly mitigates CYP-induced hepatotoxicity by reducing oxidative stress, normalizing liver enzymes, suppressing inflammation, and promoting liver regeneration. This preclinical evidence highlights the potential of CHIR99021 as an effective therapeutic agent for liver reparation and regeneration.














, Claims:CLAIMS
We Claim:
1. A composition for providing a hepatoprotective effect and promoting liver regeneration, comprising:
a GSK-3ß inhibitor, wherein the GSK-3ß inhibitor is CHIR99021; and
a pharmaceutically acceptable carrier;
wherein the composition, when administered to a subject in need thereof, inhibits GSK-3ß, leading to the accumulation of ß-catenin and activation of the Wnt/ß-catenin signaling pathway, resulting in increased hepatocyte proliferation and regeneration, and improved liver regeneration and repair.

2. The composition as claimed in claim 1, wherein the composition is formulated for administration to a subject exposed to a hepatotoxicity inducer.

3. The composition of as claimed in claim 1, wherein the hepatotoxicity inducer is cyclophosphamide.

4. The composition of as claimed in claim 1, wherein the composition is formulated for administration to a subject with liver damage.

5. The composition of as claimed in claim 1, wherein the composition is formulated for administration to a subject with a partial hepatectomy.

6. The composition as claimed in claim 1, wherein the selective GSK-3ß inhibitor is present in an amount effective to enhance cellular growth and repair in the liver.

7. A composition as claimed in claim 1, for promoting liver regeneration and providing a hepatoprotective effect, comprising:
a selective GSK-3ß inhibitor, wherein the selective GSK-3ß inhibitor is a chemical compound that inhibits GSK-3ß activity; and
one or more pharmaceutically acceptable excipients;
wherein the selective GSK-3ß inhibitor accumulates ß-catenin and activates the Wnt/ß-catenin signaling pathway in the liver, thereby stimulating liver regeneration and repair.
8. A composition as claimed in claim 1, wherein the composition is an oral tablet.

9. The composition as claimed in claim 1, wherein the solid oral composition is a tablet comprising:
about 1 mg to about 1000 mg of the selective GSK-3ß inhibitor;
about 50 mg to about 500 mg of a diluent selected from the group consisting of lactose, microcrystalline cellulose, starch, and combinations thereof;
about 5 mg to about 100 mg of a binder selected from the group consisting of polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and combinations thereof;
about 1 mg to about 50 mg of a disintegrant selected from the group consisting of croscarmellose sodium, crospovidone, sodium starch glycolate, and combinations thereof; and
about 0.1 mg to about 10 mg of a lubricant selected from the group consisting of magnesium stearate, stearic acid, talc, and combinations thereof.

10. A method for promoting liver regeneration and providing a hepatoprotective effect, comprising:
administering a composition of claim 1, comprising a selective GSK-3ß inhibitor to a subject with liver toxicity or damage, wherein the selective GSK-3ß inhibitor is a chemical compound that inhibits GSK-3ß activity;
herein the selective GSK-3ß inhibitor accumulates ß-catenin and activates the Wnt/ß-catenin signaling pathway in the liver of the subject, thereby stimulating liver regeneration and repair.
Dated this 29th October, 2024

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