Clinical trial NCT06871904: Fibrosis-Modulating Effects of Metformin and Pirfenidone in Oral Submucous Fibrosis
Official title: Exploring the Fibrosis-Modulating Potential of Metformin and Pirfenidone in Oral Submucous Fibrosis: Molecular Mechanisms to Clinical Applications
Study type: INTERVENTIONAL
Overall status: RECRUITING
Phase: PHASE1
PHASE2
Conditions: Oral Submucous Fibrosis
Keywords: Oral submucous fibrosis
Fibrosis Modulation
Metformin
Pirfenidone
Exosome secretion
Molecular Pathway
LCMS
Clinical Trial
Exosome Protein Profiling
Lead sponsor: Ziauddin University
Sponsor class: OTHER
Collaborators: name: University of Karachi
First posted: 2025-03-12
Enrollment: 45
Sex: ALL
Minimum age (years): 18
Maximum age (years): 45
Standard ages: ADULT
Accepts healthy volunteers: no
Allocation: RANDOMIZED
Intervention model: PARALLEL
Intervention model description: Part A: It will be in in vitro OSF Cell line experimental model in which exosome protein profiling and characterization will be performed .

Part B : Clinical Trial :All groups will undergo a 24-week intervention phase. The groups will be divided according to the treatment regime.

Group 1 Control (Supportive Care): Beclomethasone mouthwash three times + Capsule vitamin E 400mg daily.

Group 2 (Metformin + Supportive Care): Metformin: 500 mg daily + Beclomethasone mouthwash three times + Capsule vitamin E 400mg daily.

Group 3 (Pirfenidone + Supportive Care): Pirfenidone: 200 mg daily + Beclomethasone mouthwash three times + Capsule vitamin E 400mg daily.

Each group will be advised to refrain from spicy ingredients and cease eating pan, chalia or gutkha. All groups will be instructed to perform a mouth-opening stick exercise twice daily, alternating sides, holding the stick for 10 minutes on each side, with a 10-minute rest in between.Clinical evaluations will be conducted every 4 weeks
Primary purpose: TREATMENT

Brief summary: The goal of this clinical trial is to learn if metformin and antifibrotic drugs (pirfenidone) can modulate fibrosis and improve treatment outcomes in patients with oral submucous fibrosis (OSF). The study also aims to investigate the molecular mechanisms underlying their effects on exosome secretion and protein expression.

The main questions it aims to answer are:

Do metformin and antifibrotic drugs alter exosome secretion and biological activity in OSF cell lines? What molecular pathways are influenced by these drugs in modulating fibrosis? Does treatment with metformin and antifibrotic drugs improve clinical outcomes in OSF patients? Researchers will compare metformin and antifibrotic drug treatment groups to a control group to see if these drugs lead to significant changes in fibrosis-related exosomal protein expression and clinical improvement in OSF patients.

Participants will :

Undergo in vitro experiments on OSF cell lines to analyze drug effects using qPCR, Western Blot, and LCMS for protein profiling.

Participate in a randomized, double-blind clinical trial where they receive metformin, antifibrotic drugs, or a placebo.

Undergo clinical evaluations and laboratory tests to assess treatment efficacy. This study aims to develop an affordable and effective fibrosis-targeted therapy for OSF by repurposing metformin, potentially improving patient outcomes and reducing the risk of malignant transformation.

Detailed description: Oral submucous fibrosis stands as a persistent inflammatory and potentially malignant condition affecting the oral cavity, marked by progressive fibrosis of the oral mucosa. Existing therapies, such as corticosteroids, are costly and merely treat the symptoms without addressing the molecular pathways that cause fibrosis, which results in limited efficacy, relapse, and side effects. Given the financial constraints of affected communities, there is a desperate need for affordable and accessible treatments. This research project aims to investigate the effects of metformin and antifibrotic drugs on the secretion and biological activity of exosomes in oral submucous fibrosis (OSF) cell lines. The study will explore how these treatments impact morphological changes, exosomal gene expression, and protein profiles in OSF cell lines compared to control and vehicle-controlled groups. Additionally, the project will identify molecular pathways influenced by metformin and antifibrotic drugs, with a focus on their modulation of exosomal protein content and functional profiles. The clinical relevance of exosomal protein profiles in OSF treatment, particularly through the repurposing of metformin, will also be evaluated. Methodologically, the study involves in vitro experiments on OSF cell lines, employing techniques such as qPCR, Western Blot, and Liquid Chromatography-Mass Spectrometry (LCMS) for protein profiling, alongside a clinical trial to assess therapeutic efficacy.

The project is structured around several key objectives:

1. Impact Assessment: Evaluate how metformin and antifibrotic drugs affect exosome secretion and activity in OSF cell lines.
2. Morphological and Gene Expression Analysis: Analyze morphological changes and exosomal gene expression profiles in OSF cell lines post-treatment compared to control groups.
3. Molecular Pathways: Identify and analyze the molecular pathways modulated by metformin and antifibrotic drugs that influence exosomal protein content and functions.
4. Exosome Characterization: Characterize exosomes isolated from different treatment groups using advanced techniques such as dynamic light scattering (DLS), scanning electron microscopy (SEM), Western Blot (WB) and Liquid Chromatography-Mass Spectrometry (LCMS).
5. Clinical Relevance: Explore the potential clinical applications of the exosomal protein profiles for OSF treatment, focusing on the repurposing of metformin.
6. Methodology: The study includes both in vitro and clinical components, with the in vitro studies involving dose-response experiments, microscopy, gene expression analysis, and exosome isolation and characterization. The clinical trial will assess the efficacy of metformin and antifibrotic drugs in patients with OSF through randomized double-blind controlled trials.

This research has the potential to uncover new therapeutic strategies for OSF, particularly through the repurposing of metformin, which could lead to better clinical outcomes for patients suffering from this precancerous condition.

Eligibility criteria: Inclusion Criteria:

1. Patients with OSF having fibrous bands
2. Patients with limited mouth opening due to OSF in Stage 2 and Stage 3
3. Patients who have not received any treatment for OSF in the previous three months
4. Patients with habits of pan, Chalia, Gutkha
5. The age group between 18 and 45 years

Exclusion Criteria:

1. Patients presenting with OSCC
2. Patients with limited mouth opening due to impaction of the third molar
3. Patients with limited mouth opening due to temporomandibular joint disorder
4. Any history of Metformin intolerance or contraindications to its use.
5. Medical conditions (e.g., cardiovascular disease, renal/hepatic impairment) or drug therapy (\>6 months) with immunosuppressants, corticosteroids, or antifibrotics.
6. Pregnancy or lactation.
7. Participation in other clinical trials concurrently.
8. Inability / unwilling to provide informed written consent.

Primary outcomes:
measure: Change in Exosome Secretion and Characterization in OSF Cell Lines
timeFrame: 24, 48, and 72 hours post-treatment
measure: Changes in Fibrosis-Related Gene Expression in OSF Cell Lines
timeFrame: 24 and 48 hours post-treatment
measure: Reduction in Fibrosis Severity in OSF Patients
timeFrame: Baseline, 4 weeks, 8 weeks, and 12 weeks, 16 weeks, 20 weeks and 24 weeks
measure: Change in TGF-β and Collagen Levels in Exosomes from OSF Cell Lines
timeFrame: 24, 48, and 72 hours post-treatment
Secondary outcomes:
measure: Morphological Changes in OSF Cell Lines Post-Treatment
timeFrame: 24, 48, and 72 hours post-treatment
measure: Size and Distribution of Exosomes in OSF Cell Lines
timeFrame: 24, 48, and 72 hours post-treatment
measure: Functional Impact of Treated OSF cells
timeFrame: 24, 48, 72 and 96 hours post- treatment
measure: Safety and Viability of OSF Cell Lines Post-Treatment
timeFrame: 24, 48, and 72 hours post-treatment
Other outcomes:
Locations:
city: Karachi
country: Pakistan
facility: Ziauddin University
state: Sindh
Study officials:
References:
citation: Wang L, Zhong NN, Wang X, Peng B, Chen Z, Wei L, Li B, Li Y, Cheng Y. Metformin Attenuates TGF-beta1-Induced Fibrosis in Salivary Gland: A Preliminary Study. Int J Mol Sci. 2023 Nov 13;24(22):16260. doi: 10.3390/ijms242216260.
citation: Huh JY, Lee JH, Song JW. Efficacy and safety of combination therapy with pirfenidone and nintedanib in patients with idiopathic pulmonary fibrosis. Front Pharmacol. 2023 Dec 12;14:1301923. doi: 10.3389/fphar.2023.1301923. eCollection 2023.
citation: Chen X, Xie H, Guo J. Drug treatment for oral submucous fibrosis: an update. BMC Oral Health. 2023 Oct 12;23(1):748. doi: 10.1186/s12903-023-03488-9.
citation: Ahmadpour F, Rasouli HR, Talebi S, Golchin D, Esmailinejad MR, Razie A. Effects of exosomes derived from fibroblast cells on skin wound healing in Wistar rats. Burns. 2023 Sep;49(6):1372-1381. doi: 10.1016/j.burns.2023.02.003. Epub 2023 Feb 14.
citation: Abbasi-Malati Z, Azizi SG, Milani SZ, Serej ZA, Mardi N, Amiri Z, Sanaat Z, Rahbarghazi R. Tumorigenic and tumoricidal properties of exosomes in cancers; a forward look. Cell Commun Signal. 2024 Feb 15;22(1):130. doi: 10.1186/s12964-024-01510-3.
citation: Abbasi R, Nejati V, Rezaie J. Exosomes biogenesis was increased in metformin-treated human ovary cancer cells; possibly to mediate resistance. Cancer Cell Int. 2024 Apr 16;24(1):137. doi: 10.1186/s12935-024-03312-6.