Safety Trial of Herbal Melanin in Gastritis Patients (RASATHEME)

Open Label, Randomized, Single Site Clinical Trial To Compare The Safety Of Herbal Melanin Extracted From Nigella Sativa Seeds Vs Standard Of Care Treatment In Treating Gastritis Patients

The aim of the study is to use Melanole, a herbal extract from Nigella sativa, for treatment of gastritis. The effect of Melanole will be compared between participants (including H. pylori and non-H.pylori infected patients), with the triple therapy and Standard of care treatment of gastritis, respectively. All participants will be examined before and after the administration of Melanole. The results showing a relief of gastritis symptoms for non-H. pylori patients and partial or complete eradication of H. pylori for H.pylori infected patients will be evaluated.

Background: The melanin to be used in this study is a herbal melanin (HM) that has been extracted from the plant Nigella sativa. Nigella sativa was obtained from local market in Riyadh, Saudi Arabia. HM has been obtained from seed total powdered extract using standard extraction procedure and has been reported as a patent for the first time.The percentage of pure melanin in total Nigella sativa L. is 9% as identified using Electron Spin Resonance (ESR) and Fourier Transform- Infra Red (FI-IR) and calculated using wet chemical methods. Three different patches of Nigella sativa L that were obtained from different Nigella sativa L species were tested (Saudi, Ethiopian and Indian) and the same percentage of HM was obtained. The safety of HM has been tested and confirmed by different labs including lab for Quality and Safety and Industrial Research Institute(IRI) (see supplements 1 and 2). Similarly, the safety of the HM capsules have been tested at the Toxicology and Bioanalysis in King Faisal Specialist Hospital(KFSH) (see supplement 3). Moreover, the capsules have been approved and certified for free sale by the Ministry of Health at Lebanon. Study Area: The study will be performed in the gastroenterology clinic at National Guard Health Affairs(NGHA). Volunteers who participate in the study will be recruited from NGHA clinics. Participates are patients diagnosed with gastritis, both H. Pylori and non-H.Pylori infected, together with healthy volunteers. The histological studies will be performed at the pathology department at NGHA. Taking biopsies during endoscopy: The best way to get tissue samples from the stomach is through a procedure called an esophagogastroduodenoscopy. It is more commonly known as an endoscopy or EGD. This is generally done as an outpatient procedure. The biopsies will be taken through the esophagogastroduodenoscopy (EGD) by a ultra-thin forceps, then will be saved in special container and send to the Laboratory for staining and microscopic examination. Under the microscope the pathologist will look for the abnormal mucosa , inflammation, ulceration , and helicobacter pylori gram negative micro-organism .During the procedure there is low chance of some complications which happened in 1: 10 000 , these include ; injury to the mucosa with bleeding and perforation , discomfort , sedation related complication like headache and nausea . These rare but the patient will be informed about it. Study Subjects: Patients with gastritis or showing gastritis symptoms will be enrolled in this study. Gastritis participate patients will be selected after detection of gastric lesions by undergoing routine diagnostic tests (such as endoscopy of upper digestive system). Sample Size Calculation and Estimation: The sample size has been estimated as 132 ( 22 subjects per group). The calculation and estimation have been done as follows: Sample Size Calculation The calculation of sample size was performed using Query Advisor V.7 Sample Size Estimated The minimum number of Subjects needed in each group is 18, a two sided 95.0% confidence interval for the difference between a Group 1 proportion of 0.6 and Group 2 proportion of 0.2 based on the large sample normal approximation will extend 0.3 from the observed differences in proportions. The total required sample would be (6 groups x 18=108).The assumed withdrawal/dropout rate is 20% in each group (22 subjects per group), the final sample size would ne 132 subjects. The calculation of sample size was performed using Query Advisor V.7) Data Collection method: All data will be collected electronically and reported in Case Report Forms. Data Cleaning: Raw data will be processed in accordance with the best practices for raw data management to identify any inaccuracies or incompleteness in advance to the statistical analysis. In order to accomplish this task, all interval variables will be checked and summarized in terms of maximum and minimum values. Minimum and maximum values will be checked and compared against the nominal maximum and minimum value of each variable and variables with implausible values will be flagged. A similar process will be applied to categorical variables to identify any potential anomalies (miscode) by running a general frequency analysis. Cohort Characterization: All variables will be summarized and reported using descriptive statistics. Interval variables will be summarized and reported in terms of mean and standard deviation. Categorical variables will be summarized and reported in terms of frequency distribution. All variables will be compared across study groups using chi square and t test accordingly. Primary Outcome(s) Analysis: Chi square test will be used to compare the distribution of gastritis across the study groups. Results will be reported in terms of count, percent, and p-value. Significance will be declared at alfa less than 0.05. Binary logistic regression will be used to identify significant predictors of the primary outcomes. The statistical model will include key variables that showed significance with the primary outcome at the bivariate level. Results will be reported in terms of odds ratio, std. error, 95% confidence interval, and p-value. Significance will be declared at alfa less than 0.05. Intent to treat analysis will be the approach in analyzing the data. Statistical Analytical System(SAS) 9.2 will be used for all statistical analyses.

omeprazole40 mg P.O. Twice per Day(BID) for one month + Amoxil 1000mg P.O. BID for 2 weeks+ Clarithromycin500 mg PO BID for two weeks. Omeprazole+Amoxil+clarithromycin is the standard triple therapy given

omeprazole 40 mg P.O. BID for one month +1800 mg Herbal melanin PO TID (300mg x2 capsules) Omeprazole + Herbal melanin will be tested

Amoxil 1000mg P.O. BID for 2 weeks+ Clarithromycin500 mg PO BID for two weeks +1800 mg Herbal melanin PO TID(300 mg X 2 capsules) Herbal melanin+Amoxil+Clarithromycin will be tested

Investigators hypothesize that HM is an effective candidate that can decrease stomach acidity and eradicate H.Pylori by working as Proton Pump Inhibitor(PPI) and/or antibacterial agent, respectively. Investigators suggest the role of HM in activation of Toll like Receptor 4 (TLR4)/Cycloxygenase2(COX2)/ProstaglandinE2( PGE2) as one underlying mechanism.

Number of Participants cured with herbal melanin as compared to number of participants cured with standard of care for gastritis and H.Pylori-induced gastritis

Gastritis as indicated by improving the clinical presentation of gastritis and confirmed by end of treatment endoscopy and stool antigen test

Number of Participants between study groups having high expression of TLR4 and COX2 as assessed by Western blots procedure.

Inclusion Criteria: Males or Females between 18- 60 years Willing to sign Informed Consent Form (ICF) Women of Child bearing age will be asked to conduct a pregnancy test before enrolling in the study and offered contraceptives for the duration of the study participation Clinical Pictures of Gastritis. Histologically confirmed H- Pylori Gastritis or Non H- Pylori by one week from enrollment. Exclusion Criteria: Patients with disturbed gastrointestinal physiology (gastric surgery, vagotomy, Zollinger-Ellison syndrome) Patients who have been treated with proton pump inhibitors during 3 weeks prior to inclusion Patients who have been treated with antibiotics or bismuth containing drugs 1 month prior enrolling in the study Patient with pyloric stenosis Patient with Hematologic disorder Patient with congestive heart disease Women who are pregnant or lactating Current or past history of malignancy Drug abuser and chronic alcoholism Patients currently participating in any other clinical trial of any kind

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