Natural pharmaceutical composition for treatment of peptic ulcer

Abstract

The invention relates to a natural pharmaceutical composition for prevention and treatment of gastric ulcer and a preparation method of the natural pharmaceutical composition. The natural pharmaceutical composition comprises active natural medicinal ingredients. It is composed of turpentine oil, black seed oil, vitamin D and hyaluronic acid. The natural pharmaceutical composition provides a significant synergistic effect, combining anti-inflammatory, antibacterial, and gastroprotective properties, while also ensuring cost-effectiveness and minimal side effects compared to conventional treatments. Beside gastric ulcer is effective in relief of symptoms associated with gastric ulcer. The natural composition has the advantages of accurate curative effects, preparation stability, simplicity and feasibility in process and suitableness for industrial production.

Description

Natural pharmaceutical composition for treatment of peptic ulcer

[0001] The present invention relates to the development of a novel natural pharmaceutical composition designed for the prevention and treatment of peptic ulcer, including its preparation process and application.

[0002] Peptic ulcer disease (PUD) is characterized by discontinuation in the inner lining of the gastrointestinal (GI) tract because of gastric acid secretion or pepsin. It usually occurs in the stomach and proximal duodenum. Gastric ulcer is the most common disorder of the upper digestive tract. Gastric ulcers are a break in the mucosa of the stomach lining that penetrates to deeper layers and extends more than 5 mm in diameter. The body has natural ways to protect the stomach mucosa from the harmful acidic environment. When alterations occur to these defenses, it can lead to changes in the gastric mucosa which will eventually cause erosion and then ulceration.

[0003] The most common etiologies of gastric ulcers include a bacterial infection with Helicobacter pylori and gastric prostaglandin loss associated with non-steroidal anti-inflammatory medications. The common factor in all of these etiologies is that they promote a breakdown in the mucosal barrier and expose the gastric mucosa to the damaging effects of acid. Symptoms often include burning stomach pain, bloating, indigestion, and in some cases, nausea or vomiting.

[0004] Gastric ulcer represents a major health problem, both in terms of morbidity and mortality. Patients may be treated differently depending on the etiology of their gastric ulcer. Treatment typically involves medications to reduce stomach acid and promote healing, such as proton pump inhibitors (PPIs) or H2-receptor antagonists (H2-blockers), along with antibiotics if H. pylori is present.

[0005] The current treatments for gastric ulcer are chemically synthesized and has wide range of side effects and cannot be regularly used from the point of the safety. Therefore, for the treatment of gastric ulcer, and H. pylori, a novel natural formulation is required.

[0006] Regarding that Many patients with gastric ulcer seek natural and herbal remedies as alternative medication, numerous efforts were undertaken to develop natural formulations containing natural components along with additives for oral ingestion to treat gastric ulcer.

[0007] Nigella sativa L. (family Ranunculaceae), also known as black cumin or black seed is an annual plant. It grows mainly in selected parts of Asia and southern Europe. The beneficial properties of black cumin were already known in ancient times. It consists of non-volatile compounds, such as tannins or flavonoids, and volatile compounds, such as terpene compounds. The active compounds contained in black seed and black seed oil (BSO)show strong pharmacological potential useful for treatment of various diseases such as gastrointestinal disorders.

[0008] Turpentine is a liquid gained via distillation of oleoresin from live trees, particularly pines. Pistacia atlantica Desf is the main source of turpentine belongs to the Anacardiaceae family, and growing from the Mediterranean basin to central Asia, especially in Iran, Turkey, Iraq and Iran. It is found in the Zagros Mountains of Iran. In the sources of traditional medicine, P. atlantica oleoresin has been mentioned to have various therapeutic effects such as improving digestive disorders, eliminating inflammation, relieving upper abdominal discomfort, dyspepsia and peptic ulcer.

[0009] Hyaluronic acid (HA), which is also referred to as hyaluronan or “HA,” is a naturally occurring, water soluble polysaccharide that is a major component of the extracellular matrix and is widely distributed in animal tissues. Its biocompatibility, biodegradability, and natural occurrence appeal for its wide usage for the treatment of various human and animal diseases. HA is an important component of connective tissues, skin, vitreous humour, umbilical cord, and synovial fluid and play imperative roles in maintaining elasticity and viscosity of extracellular matrix. HA has shown wide range of pharmacological activities including anti-inflammatory, wound healing and tissue regenerating, immunomodulatory, anticancer and anti-proliferative, anti-diabetic, anti-aging, skin repairing, and cosmetic properties.

[0010] Vitamin D (also referred to as calciferol) is a fat-soluble vitamin that is naturally present in a few foods, added to others, and available as a dietary supplement. Vitamin D exists in several different forms (vitamin D1 through D5); the major forms are vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol). vitamin D3 is also produced endogenously when ultraviolet (UV) rays from sunlight strike the skin and trigger vitamin D synthesis. Vitamin D activity is associated with the modulation of a wide variety of biological systems, in addition to its roles in calcium homeostatic mechanisms. While vitamin D is well known to promote gastrointestinal calcium absorption, vitamin D also plays a role in attenuating and / or preventing the progression of several gastrointestinal diseases including Crohn’s disease, ulcerative colitis and peptic ulcer. Emerging evidence, however, has shown that hypovitaminosis D itself can negatively influence the clinical course of the gastrointestinal disease.

[0011] The invention belongs to the technical field of medicine, gastroenterology and relates to a natural pharmaceutical composition for treatment of gastric ulcer. The natural pharmaceutical composition for treatment of gastric ulcer is presented to overcome disadvantages, side effects and weak efficacies of current medicines for treating gastric ulcer and is composed of turpentine oil, black seed oil, vitamin D and hyaluronic acid.

[0012] The medicinal active components have a significant synergistic effect in treatment and prevention of gastric ulcer. Therefore, the natural pharmaceutical composition has a remarkable healing effect on gastric ulcer.

[0013] The pharmaceutical composition has unique formulation to combine novel components with enhanced bioavailability to enhance the therapeutic effect of final product.

[0014] Gastric ulcers have been acknowledged as a significant global health issue for a considerable period. The globally used and accepted first-line therapy is a triple-therapy treatment regimen that includes a proton pump inhibitor (PPI) and two antibiotics. These antibiotics may include clarithromycin and either amoxicillin or metronidazole. The second-line therapy is a quadruple-therapy treatment regimen that includes a PPI, bismuth and two antibiotics. Although conventional regimens are effective, their side effects are often inevitable and limit clinical utility.

[0015] PPIs and H2-blockers have been implicated in a number of adverse effects, leading to considerable controversies related to their long-term use. These side effects include osteoporosis, kidney injury, micronutrient deficiency, infections and gastric cancer.

[0016] Furthermore, currently treatment methods for infection with H. pylori present several concerns, the most prevalent being the rapid gain in antibiotic resistance. Recent evidences show that recurrence rates for H. pylori infection following treatment with antibiotics were found to reach over 50%.

[0017] In majority of patients who received eradication therapy for H. pylori, there are side effects and complications due to which a significant portion of patients stop treatment without completing the course of antibiotic therapy.

[0018] The widespread use of antibacterial therapy for the treatment of peptic ulcer has led to the rapid spread of H. pylori strains resistant to antibacterial drugs, which has already led to a decrease in the effectiveness of eradication therapy.

[0019] The purchase of drugs for any of the above treatment options for peptic ulcer disease in most patients causes significant financial difficulties.

[0020] The need to take several drugs for a long time adversely affect the mental state of a significant part of patients and decrease treatment compliance of patients.

[0021] Essential oils (EOs) are innate constituents of aromatic and medicinal plants which are loaded with bioactive components and metabolites that are potent enough to treat various ailments. Even though, the scope of the application of EOs is still narrow due to constraints like high hydrophobicity, poor solubility in aqueous media, uncontrolled delivery action owing to their high volatility. Micro and nano emulsion comes to the rescue of these discrepancies, with the EOs being encapsulated in it as droplets with nanometric scale diameter.

[0022] Both clinical and experimental studies have demonstrated that natural and herbal components exhibit therapeutic benefit for gastric ulcer with fewer side effects compared to conventional medicine.

[0023] Moreover, the cost of herbal medicine for gastric ulcer is only about one-sixth of that of conventional medicine.

[0024] Owing to the advantages of natural components in healthcare management, such as minimal side effects and lesser toxicity, much attention has been given to natural components for treating gastric ulcer.

[0025] Essential oils have low oral absorption due to low solubility and high first pass effect. To overcome low oral bioavailability, various lipid-based drug delivery systems and self-emulsifying systems were developed. Lipid-based delivery systems and particularly self-emulsifying delivery systems (SEDS) were demonstrated to increase the solubility, dissolution and bioavailability of many insoluble drugs.

[0026] Current invention presents a natural composition for treatment of gastric ulcer with herbal and natural components with enhanced bioavailability. Advantages of natural additives include few adverse effects, little residue, little environmental pollution, and fewer drug resistance or drug-induced diseases.

[0027] Black seed oil contains different therapeutic agents such as thymoquinone (TQ) and monoterpenes. Recent evidences show potent gastroprotective role of TQ in different models of gastric ulcer

[0028] Turpentine oil contains is considered a rich source of monoterpene compounds such as α‐pinene, β‐pinene, camphene, limonene, α‐terpineol with potent anti-inflammatory, anti-oxidant and antibacterial properties.

[0029] HA acts as a barrier to form a hydrated protective layer over ulcers, shielding the nerve endings of the mucosal tissue from external stimuli and thereby attenuating pain and shortening ulcer healing time. Furthermore, HA plays a role in cell migration and proliferation, which are essential processes in wound healing. It helps in the formation of new tissue and blood vessels. Once ingested, HA will dissolve and disperse in the stomach and form a protective membrane at the gastric ulcer region in order to accelerate healing of the inflammatory region and also to achieve sustained effect.

[0030] Vitamin D has potent antioxidant properties and can boost the expression of antioxidant enzymes, helping to reduce oxidative stress that can contribute to ulcer formation and exacerbate damage. Furthermore, Vitamin D has been shown to influence cell proliferation and differentiation in epithelial tissues, potentially promoting healing of the gastric mucosa and reducing ulcer size.

[0031] Vitamin D can induce the production of cathelicidins and defensins peptides. Cathelicidin and defensin are small peptides with amphipathic structures that allow them to disrupt the integrity of the pathogen cell membrane, resulting in its death. Both cathelicidin and defensin have activities beyond killing of microbes and can act as signaling molecules to activate and enhance the immune system.

[0032] The combination of turpentine oil, black seed oil, hyaluronic acid and vitamin D produce synergistic interaction and enhance antibacterial, anti-inflammatory, antioxidant and gastroprotective properties.

[0033] Figure list follows:Fig.1

[0034] Schematic picture depicting process of SEDS formulation and its therapeutic mechanismFig.2

[0035] The therapeutic effect of natural preparation on ulcer index in acetic acid induced gastric ulcer in rat. The control group received olive oil. The standard treatment was Amoxicillin (5 mg / kg) + Clarithromycin (25mg / kg) + Omeprazole (20 mg / kg). The treatment group received hyaluronic acid, turpentine, black seed oil and vitamin D. The asterisk indicated statistical difference with control group at P<0.05

[0036] The invention will now be described in detail in connection with certain preferred and optional embodiments, so that various aspects thereof may be more fully understood and appreciated. However, these embodiments may be considered as not self-limiting, but incorporated herein as typical examples of the invention in its various and diverse options.

[0037] The present invention in a preferred embodiment provides natural pharmaceutical compositions for prevention, improvement or treatment of gastric ulcer and associated clinical symptoms.

[0038] More specifically, the invention provides a natural composition comprising turpentine oil, black seed oil, vitamin D and hyaluronic acid.

[0039] The term natural composition for the purpose of this invention means a substance or mixture comprising of at least one natural component having medicinal property, wherein the natural component shall include a component extracted or derived from a natural source such as an herb or living organisms or from surrounding environment.

[0040] The turpentine oil for the composition is a fluid obtained by distillation and rectification from the oleoresin produced by various species of Pinus (Pinus pinaster, Pinus halepensis, …)or Pistacia (Pistacia atlantica, Pistacia atlantica subsp. Kurdica, …) genus.

[0041] The black seed oil is obtained from the seed of the genus Nigella and more specifically Nigella sativa, which is commonly known as black seed or black cumin. Plant seed oils are an excellent source of antioxidants. In addition to traditional antioxidants, such as vitamins C and E, plant seed oils contain phenolic compounds which are excellent free radical scavengers.

[0042] Vitamin D used in the current invention is intended to cover all forms of vitamin D, their derivatives and metabolites, and any combination therein.

[0043] HA as used in the present invention refers to hyaluronic acid or salts of hyaluronic acid, such as the sodium, potassium, magnesium and calcium salts, among others. The formulation HA can cover the inflammatory portion to treat the gastric ulcer and the duodenal ulcer.

[0044] The composition can be formulated to comprise turpentine oil, black seed oil, vitamin D and HA in a selected ratio. The ratio can be any ratio that is capable of treating peptic ulcer.

[0045] The composition can take any oral administration form capable of treating gastric ulcer as disclosed herein. The composition can be in the form of a liquid, capsule, solution, syrup, paste, suspension, emulsion, gel, drop, bead, beadlet, and any other form suitable for use.

[0046] The composition can be combined with a mineral, amino acid, protein, carbohydrate, lipid, fatty acid, food, beverage, nutritional or dietary supplement, excipient, pharmaceutically acceptable carrier, bulking agent, binding agent, caffeine, flavoring, sweetener, preservative, or combinations thereof.

[0047] In an embodiment of the invention, the natural composition can be prepared as an emulsion by compounding the active ingredients with the usual nontoxic pharmaceutically acceptable excipients and solvents.

[0048] In another embodiment of the invention, the natural composition can be prepared by self-emulsifying delivery system (SEDS) method to increase the water solubility of the composition, by compounding the active ingredients with the usual nontoxic pharmaceutically acceptable excipients and solvents.

[0049] The SEDS is configured to provide droplets comprising the lipophilic active ingredient. Particularly, the SEDS is configured to form a micro or nano emulsion (i.e., micelles) in the mouth and gastrointestinal tract of the patient upon use of the oral product. Providing a lipophilic active ingredient in the form of such a micro or nano emulsion may result in wide range of benefits such as: reduced lag time to systemic absorption, allow absorption via the lymphatic system, avoid first-pass metabolism, and increase the bioavailability of the lipophilic active ingredient.

[0050] Compared with classical emulsions, SEDS are physically more stable and easier to manufacture, being ready-to-use formulations to entrap lipophilic drugs with dissolution rate–limited absorption. Consequently, these systems may offer an improvement in the rate and extent of absorption, tailor pharmacokinetic profiles, and result in more reproducible blood–time profiles.

[0051] The compositions may contain inert excipients that are conventionally used in oral formulations, but are not limited to, one or more agents selected from diluents, disintegrants, solvents, cosolvents, isotonicity agents, chelating agents, buffers, pH-adjusting agents, solubilizers, stabilizers, dispersion agents, surfactants, co-surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, processing aids, colorants, sweeteners, perfuming agents, binders, flavoring agents, and other known additives.

[0052] The composition comprises at least one oil. In the context of the present invention, the term oil (lipid) refers to natural or synthetic oil in which the lipophilic compound is dissolved. The oil may be selected from mineral oil, paraffinic oils, vegetable oils, glycerides, esters of fatty acids, liquid hydrocarbons and others, as well as mixtures thereof. Examples of suitable oils that may be used in the compositions of the present disclosure include, but are not limited to, nut oils, anise oil, soybean oil, hydrogenated soybean oil, apricot kernel oil, corn oil, olive oil, peanut oil, almond oil, walnut oil, cashew oil, rice bran oil, poppy seed oil, cottonseed oil, canola oil, sesame oil, hydrogenated sesame oil, coconut oil, flaxseed oil, cinnamon oil, clove oil, nutmeg oil, coriander oil, lemon oil, orange oil, safflower oil, cocoa butter, palm oil, palm kernel oil, sunflower oil, rapeseed oil, castor oil, hydrogenated castor oil, and combinations thereof.

[0053] The formulation comprises at least one surfactant. The term surfactant refers to ionic or non-ionic surfactants, which may have a hydrophilic nature, i.e. a surfactant having an affinity for water. Examples of suitable surfactants that may be used in the compositions of the present disclosure include, but are not limited to, Tween, Caprol, Span, Cremophor, Labrasol and Triton X-100 and combinations thereof.

[0054] The formulation comprises at least one co-surfactant. The term co-surfactant refers to a chemical substance that is used in addition to a surfactant to improve its performance. Examples of suitable co-surfactants that may be used in the compositions of the present disclosure include, but are not limited to propylene glycol, ethylene glycol, polyethylene glycol, glycerol, lecithin and combinations thereof.

[0055] In an embodiment of the invention, the natural compositions as an active ingredient may be used alone or in combination with other therapeutic agents such as coagulant, antacid, H2 blocker, PPI, the potassium hydrogen ion pump blocker or the peptic mucosa protector as the adjuvant to potentiate the therapeutic effect, or combinations thereof.

[0056] Administering the composition can treat one or more symptoms associated with gastric ulcer, including, without limitation, stomach pain, abdominal pain, bloating, heartburn, nausea, vomiting, feeling full or satiated with little or no eating, belching, flatulence, burning in the stomach and abdomen, stomach growling, acidic taste, gastrointestinal dysmotility, delayed gastric emptying, and gastrointestinal gas. Administering the composition can reduce, arrest, prevent, or alleviate one or more symptoms of peptic ulcer.Examples

[0057] The present invention is explained by following examples and is not limited by these examples.

[0058] Example 1: emulsion formulation

[0059] An embodiment of the emulsion preparation was formulated as indicated in following table.

[0060] Table 1: The ingredients of emulsion formulationIngredient% by weight of emulsionSunflower oil30Turpentine15Black seed oil15Hyaluronic acid20Vitamin D0.1Lecithin10Beeswax10

[0061] Example2:SEDSformulation

[0062] An embodiment of the SEDS preparation was formulated as indicated in following table. The formulation process is showed in.

[0063] Table 2: The ingredients of SEDS formulationIngredient% by weight of SEDSTurpentine15Black seed oil15Hyaluronic acid20Vitamin D0.1Caprol15Glycerol15Deionized Water20

[0064] Example3:Preclinical study

[0065] A preclinical (experimental) study was conducted to investigate the therapeutic effect of the natural preparation containing turpentine oil, black seed oil, vitamin D and hyaluronic acid in acetic-acid induced chronic gastric ulcer in rat. The study groups included groups received different preparations, however the results of three groups are presented here. Experiments were conducted using mature Wistar strain male albino rats (190–230 g). The animals were kept in standard controlled laboratory conditions [12 h light / dark photo-cycle, temperature (22 ±2) °C] with ad libitum access to food and water. The rats were fasted 24 h prior to the start of experiments. The study was conducted in compliance with the ethical recommendation’s principles about use of laboratory animal guidelines. Briefly, animals were anesthetized with xylazine / ketamine by intraperitoneal injection. The abdomen was exposed and 500 μl of 80% (v / v) acetic acid was instilled into a glass tube barrel of 6 mm diameter that was applied to the serosal surface of the stomach for 1 min. The acetic acid was removed by aspiration, and the area was washed with sterile saline. After ulcer induction, control group received olive oil twice daily for 14 days. Positive control group received standard gastric ulcer treatment (Amoxicillin (5 mg / kg) + Clarithromycin (25mg / kg) + Omeprazole (20 mg / kg)). Treatment group was treated with herbal preparation twice daily for 14 days. At the end of the experiment, the rats were sacrificed humanely and the stomach organ was sampled for macroscopic and microscopic investigations. The ulcer index and protection rate measurement alongside histopathological investigations were performed. The data were compared between groups. The results showed that both standard treatment and the natural composition was able to reduce ulcer index and increased protection rate in treatment groups (). They were statistically different with control group that received olive oil (p<0.05). Microscopic findings showed that acetic acid caused severe disruption of gastric epithelium surface. It also showed evidence of oedema in sub mucosal layer, hemorrhage along with inflammatory infiltration. Standard treatment reduced the histological aberration induced in gastric tissue after acetic acid exposure, however it showed presence of mild inflammatory infiltration and oedema. In treatment group, the morphological structure of the gastric mucosa tended to normalize, but a small amount of lymphocyte infiltration was still observed in the gastric mucosa as indicated in the following table.

[0066] Table 3. Histological changes observed in different study groups.GroupsNecrosisHemorrhageInflammationCongestionOedemaCellular infiltrationControl+++++++++++++++++++Standard+-+-++Treatment-+++++

[0067] The present invention is particularly suitable for application in the traditional, herbal and complementary medicine industries providing new methods of treatment of diseases. The new composition provided here can be commercially marketed in these industries.

[0068] The Citation List follows:

[0069] The US patent US20110166099A1, disclosed a mixture of two hyaluronic acids with different molecular weight for treatment of peptic and duodenal ulcer and bleeding.

[0070] The European Patent EP1017403B1, disclosed the use of zinc hyaluronate against peptic ulcer. The compositions employed zinc hyaluronate as an active ingredient having a molecular weight in the range of 500,000 to 1,200,000 daltons.

[0071] Tavakkoli A, Mahdian V, Razavi BM, Hosseinzadeh H. Review on clinical trials of black seed (Nigella sativa) and its active constituent, thymoquinone. J pharmacopuncture. 2017;20(3):179.

[0072] Abdi S, Abazarikia A, Mojab F, Zahedi A, Moghtaderi M, Mortazavi E, et al. Efficacy and safety of alpha-pinene capsule in the management of functional dyspepsia and eradication of helicobacter pylori: a randomized clinical trial. Gastroenterol Hepatol From Bed to Bench. 2024;17(2):140.

[0073] Alizadeh‐naini M, Yousefnejad H, Hejazi N. The beneficial health effects of Nigella sativa on Helicobacter pylori eradication, dyspepsia symptoms, and quality of life in infected patients: A pilot study. Phyther Res. 2020;34(6):1367–76.

[0074] Eftekharafzali M, Mehrabani M, Tajadini H, Ahmadi B, Zahedi MJ. Effect of “Pistacia atlantica” resin (baneh) on functional dyspepsia: A double-blind, randomized clinical study. Iran Red Crescent Med J. 2018;20(7).

[0075] Afrasiabian H, Imanieh MH, Nejati MA, Salehi A. The effect of oral administration of pistacia atlantica kurdica gum on the eradication of helicobacter pylori in patients with dyspepsia: A randomized clinical trial. Glob J Health Sci. 2016;9(2):256–63.

[0076] Hashem-Dabaghian F, Agah S, Taghavi-Shirazi M, Ghobadi A. Combination of Nigella sativa and honey in eradication of gastric Helicobacter pylori infection. Iran Red Crescent Med J. 2016;18(11):14–9.

[0077] Mohtashami R, Huseini HF, Heydari M, Amini M, Sadeqhi Z, Ghaznavi H, et al. Efficacy and safety of honey based formulation of Nigella sativa seed oil in functional dyspepsia: A double blind randomized controlled clinical trial. J Ethnopharmacol. 2015;175:147–52.

[0078] Voigt J, Driver VR. Hyaluronic acid derivatives and their healing effect on burns, epithelial surgical wounds, and chronic wounds: A systematic review and meta‐analysis of randomized controlled trials. Wound repair Regen. 2012;20(3):317–31.

[0079] Bayır Y, Turan C, Karagöz Y, Albayrak A, Erkayman B, Duysak L. Thymoquinone Ameliorates Indomethacin-Induced Gastric Ulcers in Rats: A Dose Response Study. New Trends Med Sci. 2021;2(1):15–23.

[0080] Karampour NS, Arzi A, Rezaie A, Ghasemi F, Pashmforosh M. Gastroprotective Effects of Pistacia atlantica Extract on Ethanol-Induced Gastric Ulcers in Rats. Jundishapur J Nat Pharm Prod. 2019;14(1).

[0081] Paseban M, Niazmand S, Soukhtanloo M, Meybodi NT. The preventive effect of Nigella sativa seed on gastric ulcer induced by indomethacin in rat. J Herbmed Pharmacol. 2020;9(1):12–9.

[0082] Memariani Z, Sharifzadeh M, Bozorgi M, Hajimahmoodi M, Farzaei MH, Gholami M, et al. Protective effect of essential oil of Pistacia atlantica Desf. on peptic ulcer: Role of α-pinene. J Tradit Chinese Med. 2017;37(1):57–63.

[0083] Hashem-Dabaghian F, Agah S, Taghavi-Shirazi M, Ghobadi A. Combination of Nigella sativa and honey in eradication of gastric Helicobacter pylori infection. Iran Red Crescent Med J. 2016;18(11):14–9.

[0084] Al-Bayaty F, Abdulla M, Darwish P. Evaluation of hyaluronate anti-ulcer activity against gastric mucosal injury. African J Pharm Pharmacol. 2011;5(1):23–30.

[0085] Kanter M, Demir H, Karakaya C, Ozbek H. Gastroprotective activity of Nigella sativa L oil and its constituent, thymoquinone against acute alcohol-induced gastric mucosal injury in rats. World J Gastroenterol WJG. 2005;11(42):6662.

[0086] Shojaeian A, Khalilian A, Mehrpoya M, Poorolajal J, Dastan D. Efficacy of a standardised herbal formulation in Helicobacter pylori positive patients with gastritis or peptic ulcer: a triple-blind randomised clinical trial. J Herb Med. 2023;41:100706.

[0087] Dabos KJ, Sfika E, Vlatta LJ, Frantzi D, Amygdalos GI, Giannikopoulos G. Is Chios mastic gum effective in the treatment of functional dyspepsia? A prospective randomised double-blind placebo controlled trial. J Ethnopharmacol. 2010;127(2):205-9.

[0088] Iannitti T, Morales-Medina JC, Merighi A, Boarino V, Laurino C, Vadalà M, et al. A hyaluronic acid-and chondroitin sulfate-based medical device improves gastritis pain, discomfort, and endoscopic features. Drug Deliv Transl Res. 2018;8:994–9.

[0089] Das DS, Saharia GK, Panigrahi MK, Sahoo D. Association of vitamin D and functional dyspepsia: a case-control study. Ann Med Surg. 2023;85(10):4667–73.

[0090] Yang L, He X, Li L, Lu C. Effect of vitamin D on Helicobacter pylori infection and eradication: A meta‐analysis. Helicobacter. 2019;24(5):e12655.

[0091] Albatineh AN, Sohrabi R, Baghi V, Ghanei Gheshlagh R. Effect of vitamin D on Helicobacter pylori infection and eradication: An updated systematic review and meta-analysis. J Prev Complement Med. 2023;2(3):155–62

Claims

A natural pharmaceutical composition for treatment of gastric ulcer, wherein the composition is consisting of turpentine oil, black seed oil, vitamin D and hyaluronic acid along with one or more pharmaceutically acceptable additives / carriersThe composition of claim 1, wherein the molecular weight of hyaluronic acid is between 500 and 2000 kDa.According to claim 1, emulsion (water in oil) or self-emulsifying delivery system (SEDS) formulation were prepared during a multi-step process of product formulation.The composition of claim 3, wherein at least one oil, one surfactant and one co-surfactant is used for emulsion and SEDS formulation.The composition of claim 3, wherein a food grade oil is used as a lipid base of the formulation.According to claim 3, the aqueous phase of emulsion was prepared by mixing hyaluronic acid with glycerol between 1-4 hours in 25-50 ºC.According to claim 3, the oil phase of emulsion was prepared by mixing turpentine oil, black seed oil, vitamin D between 1-4 hours in 25-50 ºC.According to claim 3, the emulsion was prepared by gradual adding of mixture obtained from claim 10 to the mixture obtained from claim 11 and mixing between 1-4 hours in 25-50 ºC.According to claim 3, turpentine, black seed oil and vitamin D were mixed in 25-50 ºC for 1-4 hours for preparation of oil phase of SEDS formulation.According to claim 3, hyaluronic acid, surfactant and co-surfactant were mixed in 25-50 ºC for 1-4 hours for preparation of aqueous phase of SEDS formulation.According to claim 3, the mixture obtained from claim 9 was gradually added to the mixture obtained from claim 10 for 1-4 hours in 25-50 ºC for preparation of SEDS formulation.According to claim 10, the ratio of surfactant to co-surfactant was between 1:9 to 9:1

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