CA2656220A1 - Compositions and methods for treating and preventing gastro esophageal reflux disease - Google Patents

Abstract

The present invention relates to methods and compositions effective in treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD). The compositions comprise, as an active ingredient, an extract from the plant genus Conyza Less, preferably from the species Conyza bonariensis (L) Cronq. The composition comprises only natural ingredients and has been demonstrated to be safe for use, while still providing a beneficial therapeutic effect. The composition can also be administered in combination with traditional anti-GERD medicines, either over-the-counter or prescription medicines such as antacids, H2-receptor agonists, proton pump inhibitors, promotility agents and Sucralfate. The compositions are particularly effective in treating and preventing GERD, the symptoms or GERD and conditions associated with GERD, including heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.

Description

COMPOSITIONS AND METHODS FOR TREATING AND PREVENTING
GASTRO ESOPHAGEAL REFLUX DISEASE

FIELD OF THE INVENTION
The present invention relates to pharmaceutical compositions comprising an extract from the plant genus Conyza Less., and preferably from the species Conyza bonariensis (L) Cronq, and their use in treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD) and associated conditions.

BACKGROUND OF THE INVENTION
Gastro esophageal reflux disease (GERD) is a condition in wliich the acids from the stomach move backward from the stomach into the esophagus (an action called reflux).
Reflux occurs if the muscular actions in the esophagus or other protective mechanisms fail.
The hallmark symptoms of GERD are heartburn - a burning sensation in the chest and throat;
and regurgitation -a sensation of acid backed up in the esophagus. Although acid is a primary factor in damage caused by GERD, other products of the digestive tract, including pepsin and bile, can also be harmful.
More than 60 million adults in the United States suffer from GERD and heartburn at least once a month, according to the National Digestive Diseases Information Clearinghouse (NDDIC). About 25 million men and women experience daily heartburn. Recent studies report that infants and children also are affected by GERD. Their symptoms include failure to thrive, recurrent vomiting, coughing, and respiratory problems. Several conditions are associated with GERD, including Esophagitis (inflammation or irritation of the esophagus), Non-Erosive Esophageal Reflux Disease (NERD) (symptoms of GERD occurring without any signs of inflammation or injury to the esophagus), and Barrett's Esophagus (a serious fonn of GERD that can lead to cancerous changes in the tissue lining of the esophagus).
The esophagus, commonly called the food pipe, is a narrow muscular tube about nine and a half inches long. It begins below the tongue and ends at the stomach.
The esophagus is narrowest at the top and bottom; it also narrows slightly in the middle. The esophagus consists of three basic layers: 1) an outer layer of fibrous tissue; 2) a middle layer containing smoother muscle; and 3) an inner membrane, which contains numerous tiny glands. When a person swallows food, the esophagus moves it into the stomach through the action of peristalsis, wave-like muscle contractions. In the stomach, acid and various enzymes, notably hydrochloric acid and pepsin, break down the starch, fat, and protein in food. The stomach has a thin layer of mucus that protects it from these fluids. If acid and enzymes back up into the esophagus, its lining offers only a weak defense.
The esophagus is protected from using specific muscles and other factors. The most important structure protecting the esophagus may be the lower esophageal sphincter (LES).
The LES is a band of muscle around the bottom of the esophagus where it meets the stomach. The LES opens after a person swallows to let food enter the stomach and then inunediately closes to prevent regurgitation of the stomach contents, including gastric acid.
The LES maintains this pressure barrier until food is swallowed again. If the pressure barrier is not sufficient to prevent regurgitation and acid backs-up (reflux), then peristaltic action of the esophagus serves as an additional defense mechanism and pushes the contents back down into the stomach.
Current treatment protocols for mild symptoms of GERD, experienced most often after eating, include prescribing over-the-counter antacids, such as Mylanta, or Tums or over-the-counter acid relievers, also known as H2-receptor agonists including Pepcid, Tagamet, or Zantac. These medicines work by neutralizing stomach acids or by forming foam at the top of the stomach as a barrier to the acid.
If symptoms are more severe, prescription drugs may be given. Examples of such drugs include H2-receptor agonists (can be given in prescription or OTC
strengths) proton pump inhibitors, promotility agents and Sucralfate (an anti-ulcer drug). H2-receptor agonists (also known as H2 blockers) are usually given to patients with mild to moderate symptoms and are especially beneficial in treating episodic symptoms. These drugs work by suppressing acid production in the stomach. H2-receptor agonists have been found to be more effective than proton-pump inhibitors at relieving symptoms that occur at night.
Proton pump inhibitors have been found to heal erosive esophagitis more rapidly than H2 blockers. These drugs also have been found to significantly improve syinptoms and signs of esophageal mucosal damage. Promotility agents (e.g., Propulsid) are used to increase the rate at which the stomach is emptied. They work by increasing pressure on the LES to help food move more quickly from the stomach. However, because of evidence regarding serious complications related to interactions with other coinmonly prescribed medications, promotility agents were removed from the market in March 2000. Now, they are only available through a limited access program. Sucralfate also may be suggested to allow the esophagus to heal or to prevent disease complications from developing. This medication works by coating the stomach to provide a barrier against stomach acid.
Current treatment drugs, whether over-the-counter or physician prescribed are, however, costly and are not free of side effects. There remains an unmet need in the art for alternative compounds and compositions, preferably of a natural source, for preventing and treating GERD, which are safe and effective and have reduced side effects compared to current treatment methods.
Conyza bonariensis (L.) Cronq is a plant species which grows systematically along path- and road-sides, also in disturbed land and neglected flowerbeds. It has a stem which grows up to 40 cm, and it branches out in long branches for the inflorescences. The leaves are long and narrow, with some teeth along the margins, and they are covered in hair. The capitula are small; the flowers have small ligules. When it bears fruit it forms a lot of achenes with white pappi. It flowers during summer and in autumn. This annual or biennial species is naturalized from the American tropics and has become a pest in southern California, and in many localities in the northern part of the State.
Synonyms for Conyza bonariensis (L.) Cronq include Erigeron bonariensis L.;
Erigeron crispus Pourr.; Erigeron linifolius Willd.; Leptilon bonariense (L.) Small; and Leptilon lin.ifolium (Willd.) Small. The species belongs to the Conyza Less.
genus, which is part of the Tracheobionta (Vascular plants) Subkingdom; the Spermatophyta (seed plants) Superdivision; the Magnoliophyta (Flowering plants) Division; the Magnoliopsida -Dicotyledons Class; the Asteridae Order and the Asteraceae - Aster Family.
Other members of the Conyza genus include Conyza canadensis (L.) Cronq. (Canadian horseweed); Conyza canadensis (L.) Cronq. var. canadensis - (Canadian horseweed); Conyza canadensis (L.) Cronq. var. glabrata (Gray) Cronq. - (Canadian horseweed); Conyza canadensis (L.) Cronq.
var. pusilla (Nutt.) Cronq. - (Canadian horseweed); Conyzafloribunda Kunth -(asthmaweed); Conyza laevigata (L.C. Rich) Pruski - (manzanilla horseweed);
Conyza pYimulifolia (Lam.) Cuatrec. & Lourteig - (primroseleaf horseweed); and Conyza namosissima Cronq. - (dwarf horseweed).
Medicinally, Conyza bonariensis (L.) Cronq,\ is a pungent tonic, astringent and diuretic, claimed to be efficient in diarrliea, kidney stones, diabetes, Urinary tract infections and hemorrhage of the bowels, uterus and of wounds. It is also recommended for pimples.
Other members of this plant family possess these medicinal properties, to various degrees.

SUMMARY OF THE INVENTION
The present invention relates to methods and compositions effective in treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD). The compositions comprise, as an active ingredient, an extract from the plant genus Cofzyza Less, preferably from the species Coflyza bonariensis (L) Cronq. The coinposition comprises only natural ingredients and has been demonstrated to be safe for use, while still providing a beneficial therapeutic effect. The composition can also be administered in combination with traditional anti-GERD medicines, either over-the-counter or prescription medicines such as antacids, H2-receptor agonists, proton pump inhibitors, promotility agents and Sucralfate. The compositions are particularly effective in treating and preventing GERD, the symptoms or GERD and conditions associated with GERD, including heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof:
The compositions of the present invention offer significant advantages over current state-of-the-art GERD treatment options, as they contain only natural ingredients and are thus safe for use. Therefore, use of the compositions of the invention will reduce or completely eliminate the unpleasant side effects typically associated with traditional or conventional anti-GERD medicines as further discussed below, while still achieving a beneficial therapeutic effect. Therefore, patient compliance may be substantially improved.
Accordingly, the present invention provides a significant improvement over the current-state-of-the-art pertaining to GERD treatment.
The present invention thus provides, in one embodiment, a method of treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, by administering to a subject in need thereof a composition comprising, an extract from a Conyza Less plant, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD. Symptoms and conditions associated with GERD include, but are not limited to, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
In another aspect, the present invention provide pharmaceutical compositions useful for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, the compositions comprising an extract from a Coizyza Less. plant, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD; and a pharmaceutically acceptable carrier or excipient.
In another aspect, the present invention relates to the use of an extract from a Conyza Less. plant, in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD.
In another aspect, the present invention relates to the use of an extract from a Conyza Less. plant the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
Any species of mixture or species from the Conyza Less. plant genus can be used in the present invention. Examples of suitable species include, but are not limited to: Conyza bonariensis (L) Cronq. (asthma weed), Conyza canadensis (L.) Cronq. (Canadian horseweed); Conyza canadensis (L.) Cronq. var. canadensis - (Canadian horseweed);
Conyza canadensis (L.) Cronq. var. glabrata (Gray) Cronq. - (Canadian horseweed);
Conyza canadensis (L.) Cronq. var. pusilla (Nutt.) Cronq. - (Canadian horseweed); Conyza floribunda Kunth - (asthmaweed); Conyza laevigata (L.C. Rich) Pruski -(manzanilla horseweed); Conyza primulifolia (Lam.) Cuatrec. & Lourteig - (primroseleaf horseweed);
and Conyza ramosissima Cronq (dwarf horseweed).
In a currently preferred embodiment, an extract from a Conyza bonariensis (L) Cronq species is used. In accordance with this preferred embodiment, the present invention provides a method of treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, by administering to a subject in need thereof a composition comprising an extract from the plant species Conyza bonariensis (L) Cronq, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD. Symptoms and conditions associated with GERD include, but are not limited to, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
In another aspect, the present invention provide pharmaceutical compositions useful for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, the compositions comprising an extract from the plant species Conyza bonariensis (L) Cronq, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated witli GERD; and a pharmaceutically acceptable carrier or excipient.
In another aspect, the present invention relates to the use of an extract from the plant species Conyza bonariensis (L) Cronq, in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD.
hl another aspect, the present invention relates to the use of an extract from the plant species Conyza bonariensis (L) Cronq in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
The term "Gastro Esophageal Reflux Disease (GERD)" includes all individuals who are exposed to the risk of physical complications from gastro-esophageal reflux, or who experience clinically significant impairment of health-related well-being (quality of life), due to reflux-related symptoms, after adequate reassurance of the benign nature of their symptoms.
In one enibodiment, the extract has anti-oxidant activity.
Though not wishing to be bound by any particular mechanism or theory, it is conteinplated that the plant extract of the present invention is effective at treating the symptoms of GERD, by affecting the function of the lower esophagel sphincter (LES), so that food and digestive juices in the stomach cannot wash back into the esophagus. The extract further normalizes stomach and intestine functions, thereby leading to relief in symptoms such as bloating, spasmodic, vomiting, diarrhea and gas.
In one embodiment, the extract is a solid, dry extract. In another embodiment, the extract is an aqueous (water) extract. In another embodiment, the extract is an alcoholic extract. The extract is provided in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD. The recommended daily dose is varying according to the degree of GERD severity and the patient undergoing treatment, where the optimal effective and safe range can typically range from about 10 - 2000 mg of dried extract, preferably from about 50 to about 2000 mg of dried extract, and more preferably from about 50 to about 500 of dried extract.
Currently preferred dosages are about 50 mg, about 100 mg, and about 150 mg of dried extract.

The composition typically comprises from about 10% to about 90% of the plant extract, for example from about 20% to about 80% plant extract, from about 30%
to about 70% plant extract. Currently preferred concentration of plant extract is about 40% or about 50%. All percentages are expressed herein as weight percentages based on the total weight of the composition.
In some embodiments, the Conyza Less (e.g. Conyza bonariensis (L) Cronq) extract of the present invention can be taken alone, for example when the symptoms of GERD are noticed, or on a regular regimen as a natural supplement. In other embodiments, however, the extract can be taken together with the one or more additional active ingredients which are traditional anti-GERD medicines, either over-the-counter or prescription medicines.
Examples of such agents include, but are not limited to, antacids, H2-receptor agonists, proton pump inhibitors, promotility agents, sucralfate, and any combination thereof. The extract and the at least one other agent can be formulated in the saine composition, or they can be in different compositions. If administered in separate compositions, the extract and the at least one other agent cain be administered sequentially or concurrently, in any order. It is contemplated that the concurrent use of the plant extract of the present invention together with the traditional anti-GERD medicines, will lower the dose of the traditional medicines that is required in order to reach a therapeutic effect, resulting in decreased side effects and higher patient compliance.
The administration schedule will depend on several factors such as the patient population, age, sex, the severity of the symptoms, etc. For example, the compositions of the invention can be taken once-daily, twice-daily, thrice daily, once-weekly or once-monthly, or the composition can be taken on an as-needed regimen to relieve the symptoms of GERD as needed. The composition is preferably taken before a meal, but can also be taken after meal or in-between meals, as needed.
The present invention further relates pharmaceutical compositions comprising a Conyza Less extract, and at least one pharmaceutically acceptable carrier or excipient. In another embodiment, the present invention relates to pharmaceutical compositions comprising a Conyza bonariensis (L) Cronq extract, and at least one pharmaceutically acceptable carrier or excipient. Examples of excipients or carriers include, but are not limited to, fillers, diluents, binders, disintegrating agents, surfactants, lubricants, solvents, stabilizers, solubilizers, tonicity enhancing agents, buffers, preservatives, thickeners, complexing agents, emulsifiers, anti-oxidants, flavor-improving agents, gellants, glidants, sweeteners, colorants, viscosity-increasing agents, and any combinations thereof. A currently preferred excipient is a calcium phosphate such as tricalcium phosphate (TCP). In a specific embodiment, the composition comprises a Conyza bonariensis (L) Cronq extract and TCP. In another embodiment, the composition further comprises at least one vitamin, e.g., vitamin E.
The compositions of the invention can be administered via oral, topical, sublingual, buccal, parenteral, paracanceral, intravenous, transdermal, inhalation, intranasal, vaginal, intramuscular, transmucosal, transdermal, rectal, intradermal, subcutaneous, intraperitoneal, intraventricular, intracranial, rectal, vaginal, or intratumoral administration. Suitable oral administration forms include, but are not liinited to, a tablet, caplet , capsule (e.g., soft or hard gelatin capsule), microcapsule, pellet, pill, powder, syrup, gel, slurry, granule, suspension, dispersion, emulsion, liquid, solution, dragee, bead and beadlet.
The compositions of the present invention are natural compositions that are higllly effective in treating, preventing, reducing the severity of, or relieving the symptoms of GERD and associated conditions such as heartburn, regurgitation, esophagitis, NERD and Barrett's Esophagus. The compositions are safe to use and are particularly advantageous due to the reduction or elimination of side effect that are associated with conventional anti-GERD
drugs currently on the market.
Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE FIGURES
The present invention will be understood and appreciated more fully from the following detailed description taken in conjunction with the appended figures which depict:
FIGURE 1: LDH test. Human Fibroblast cells were incubated with different amounts of Conyza bonariensis (L) Cronq extract for 24 h (Figure 1A), 48 h (Figure 1B), and 72 h (Figure 1C). At the end of the incubation time, LDH activity was monitored following oxidation of NADH as the decrease in absorbance at 334 nm.
The percentage of LDH released is defined as the ratio of LDH activity in the supernatant to the sum of LDH amount released plus the activity measured in the cell lysate. The figures depict the LDH concentration (arbitrary units) as a function of extract concentration (mg/ml).

FIGURE 2: Antioxidant effect of Conyza bonariensis (L) Cronq extract. Rat liver homogenates were incubated with 100 M FeSO4 as reactive oxygen species (ROS) generating system witli various concentrations of the extract. The extent of lipid peroxidation was measured by detecting malondialdehyde (MDA), an end product of peroxidative decomposition of polyeonic fatty acids. Results are plotted as the MDA
concentration (mmol) /mg protein vs. extract concentration.

FIGURE 3: Mitochondrial toxicity testing (MTT). A co-culture of human hepatocytes (HepG2) and monocytes (THP1) was treated with various concentrations of Conyza bonariensis (L) Cronq extract for 24 hours. Cells were then assayed for mitochondrial function by following the conversion of the yellow dye, tetrazolium bromide, to the red formazan derivative by mitochondrial succinate dehydrogenase.
The results are plotted as the %MTT formazan in control cells and in cells treated with 10 and 100 g/ml plant extract.

FIGURE 4: Change in efficacy parameters. Patients were treated with Conyza bonariensis (L) Cronq extract. The following end points were evaluated: (a) Change in Integrated Well-Being Index (WBI) on a 1-10 scale (a combination of Gastrointestinal Symptom Rating Scale (GSRS) and the Psychological General Well-Being (PGWB) indexes); (b) change in the Extra-esophageal symptoms recording (EES); and (c) change in the severity of the clinical symptoms.

FIGURE 5: Scheme depicting extraction procedure from Conyza boriariensis (L) Cronq.

FIGURE 6: HPLC chromatograms and US VIS spectrums of main peaks of Conyza bonariensis (L) Cronq extracts. Figures 6A and 6 B depict water extract 100%.
Figures 6C-6F depict an alcohol extract 99%. Figures 6G-6J depict an alcohol extract 50%. Figures 6K and 61 depicts the standard Apigenin.

DETAILED DESCRIPTION OF THE PRESENT INVENTION
The present invention relates to methods and compositions effective in treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD). The compositions comprise, as an active ingredient, an extract from the plant genus Conyza Less, preferably from the species Conyza bonariensis (L) Cronq. The composition comprises only natural ingredients and has been demonstrated to be safe for use, while still providing a beneficial therapeutic effect. The compositions are particularly effective in treating and preventing GERD, the symptoms or GERD and conditions associated with GERD, including heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
The present invention thus provides, in one embodiment, a method of treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, comprising administering to a subject in need thereof a composition comprising, an extract from a Conyza Less plant, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD. Symptoms and conditions associated with GERD include, but are not limited to:
heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof In another embodiment, the present invention provides a method of treating, preventing, reducing the severity of, or relieving the symptoms of heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof, comprising administering to a subject in need thereof a composition comprising, an extract from a Conyza Less plant, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
In another aspect, the present invention provide pharmaceutical compositions useful for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, the compositions comprising an extract from a Conyza Less plant, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD; and a pharmaceutically acceptable carrier or excipient.

In another aspect, the present invention relates to the use of an extract from a Conyza Less plant, in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD.
In another aspect, the present invention relates to the use of an extract from a Conyza Less. plant the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
Any species of inixture or species from the Conyza Less. plant genus can be used in the present invention. The Conyza Less. plant genus comprises six species and three varieties. The six species are: 1) Conyza bonariensis (L) Cronq (asthma weed), 2) Conyza canadensis (L.) Cronq. (Canadian horseweed); 3) Conyza f oYibunda Kunth -(asthmaweed);
4) Conyza laevigata (L.C. Rich) Pruski - (manzanilla horseweed); 5) Conyza pYimulifolia (Lam.) Cuatrec. & Lourteig -(primroseleaf horseweed); and 6) Conyza ramosissima Cronq (dwarf horseweed). The Conyza canadensis (L.) Cronq. species includes three varieties: a) ' Conyza canadensis (L.) Cronq. var. canadensis - (Canadian horseweed); b) Conyza canadensis (L.) Cronq. var. glabrata (Gray) Cronq. -(Canadian horseweed); and c) Conyza canadensis (L.) Cronq. var. pusilla (Nutt.) Cronq. - (Canadian horseweed);
In a currently preferred embodiment, an extract from a Conyza bonariensis (L) Cronq species is used. In accordance with this preferred embodiment, the present invention provides a method of treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD, comprising administering to a subject in need thereof a composition comprising an extract from the plant species Conyza bonariensis (L) Cronq, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD.
In another embodiment, the present invention relates to a method of treating, preventing, reducing the severity of, or relieving the symptoms of heartbunz, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any coinbination thereof, by administering to a subject in need thereof a composition comprising an extract from the plant species Conyza bonariensis (L) Cronq, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.

In another aspect, the present invention relates to the use of an extract from the plant species Conyza bonariensis (L) Cronq, in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of GERD and conditions associated with GERD.
In another aspect, the present invention relates to the use of an extract from the plant species Conyza bonariensis (L) Cronq in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, heartburn, regurgitation, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.
The compositions of the present invention contain only natural ingredients and are thus safe for use. Therefore, use of the compositions of the invention will reduce or completely eliminate the unpleasant side effects typically associated with traditional or conventional anti-GERD medicines as further discussed below, while still achieving a beneficial therapeutic effect. Therefore, patient compliance may be substantially improved.
Accordingly, the present invention provides a significant improvement over the current-state-of-the-art pertaining to GERD treatment.
The plant extract can be provided a solid, dry extract or in a liquid form.
The extract can be provided alone, or mixed with one or more inert excipients. In one embodiment, the extract is provided as a solid (dry) extract. In another embodiment, the extract provided in a liquid form. In another embodiment, the extract is provided as an aqueous (water) extract. In another embodiment, the extract is provided an alcoholic extract. The final product, whether solid or liquid, can comprise from about 1% to about 100% extract, the balance being made up of inert excipients. For example, the product can comprise from about 10%
to about 90%
extract, from about 20% to about 80% extract, from about 30% to about 70%
extract, from about 40% to about 60% extract, and the like. In one currently preferred embodiment, the product comprises about 40% extract and about 60% inert excipient(s). In another currently preferred embodiment, the product comprises about 50% extract and about 50%
inert excipient(s). All percentages recited herein refer to weight/weight (wt/wt) percentages, based on the total weight of the composition.
The extract is provided in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD. The dose depends on factors the severity of the condition being treated, and on the parameters of the patient (such as age, sex etc.) For example, the composition can comprise from about 10 mg to about 2000 mg extract, preferably from 50 mg to about 2000 mg extract, and more preferably from about 50 to about 500 of extract. Currently preferred dosages are about 50 mg, about 100 mg, and about 150 mg of extract. It is understood by a person of skill in the art that the above doses are provided based on the weight of a dry extract.
The administration schedule will depend on several factors such as the patient population, age, sex, the severity of the symptoms, etc. For example, the compositions of the invention can be taken once-daily, twice-daily, thrice daily, once-weekly or once-monthly, or the composition can be taken on an as-needed regimen to relieve the symptoms of GERD as needed. For example, the patient may choose to take the composition only when experiencing symptoms of GERD, such as heart bum or regurgitation, and may choose to stop taking the extract when symptoms have ceased. Alternatively, the composition can be taken on a regular basis before or after meals, to prevent the onset of GERD
and/or relieve the symptoms thereof. In yet another embodiment, the compositions can be taken on a regular basis as a dietary supplement, e.g., once-a-day.
Combination Treatments The compositions of the present invention, in one embodiment, comprise the Conyza Less (e.g. Conyza bonariensis (L) Cronq) extract as a sole active ingredient.
In another embodiment, however, the compositions can further comprise conventional or traditional anti-GERD medicines or drugs.
The term "traditional or conventional anti-GERD medicines or drugs", as used herein, includes both over-the-counter (OTC) drugs as well as prescription medicines.
These medicines work by neutralizing stomach acids or by forming foam at the top of the stomach as a barrier to the acid. Examples of such medicines include, but are not limited to:
Antacids: Alka-Seltzer, Maalox, Mylanta, Pepto-Bismol, Rolaids, Riopan, and Tums are the first drugs recommended to relieve heartburn and other mild GERD
symptoms. These are OTC medications. They use combinations of three basic salts--magnesium, calcium, and aluminum--with hydroxide or bicarbonate ions to neutralize stomach acids.
There are side effects of these medicines. Magnesium containing preparations can cause diarrhea; aluminum salts can cause constipation. Aluminum and magnesium salts are often combined in a single product to balance these effects. Calcium containing antacids can also be a supplemental source of calcium but they can also cause constipation.

H2 Receptor antagonists (H2RAs): The most popular brands of this type of acid reflux medication are approved for OTC use. The four OTC H2RAs - ranitidine (Zantac), cimetidine (Tagamet), famotidine (Pepcid), and nizatidine (Axid) suppress acid production.
These GERD medicines provide short-term relief. The OTC preparations should not be used for more than a few weeks at a time. The prescription strength preparations may be taken longer, simply because they are taken under the supervision of a doctor. H2RAs work for about 50% of those patients with GERD symptoms.
Proton pump inhibitors: Proton pump inhibitors are available by prescription only and are usually given to patients after H2RAs have failed. These drugs include omeprazole (Prilosec), lansoprazole (Prevacid), pantoprazole (Protonix), rabeprazole (Aciphex), and esomeprazole (Nexium). Proton pump inhibitors are more effective than H2 RAs and relieve symptoms in almost everyone who has GERD.
Promotility agents (Propulsid) are used to increase the rate at which the stomach is emptied. They work by increasing pressure on the LES to help food move more quickly from the stomach. However, because of evidence regarding serious complications related to interactions with other commonly prescribed medications, promotility agents were removed from the market in March 2000. Now, they are only available through a limited access program.
Sucralfate also may be suggested to allow the esophagus to heal or to prevent disease complications from developing. This medication works by coating the stomach to provide a barrier against stomach acid.
Conventional anti-GERD medicines, whether OTC or prescription drugs, are associated with unpleasant side effects which at times can be severe and may lead to reduced patient compliance. For example, side effects of Zantac include an allergic reaction (difficulty breathing; closing of your throat; swelling of lips, tongue, or face; or hives), easy or unusual bruising or bleeding, bleeding gums, irregular heartbeat, yellowing of the skin or eyes, dizziness, headache, diarrhea, nausea or constipation. In addition, Mylanta antacids contain aluminum and magnesium salts. Magnesium salts may cause diarrhea, while aluminuin salts may cause constipation. Side effects associated with Tagamet include breast development in men, headache, agitation, anxiety, confusion, depression, disorientation, and hallucinations, and rare side effects include allergic reactions, anemia, blood disorders, diarrhea, dizziness, fever, hair loss, impotence, inability to urinate, joint pain, kidney disorders, liver disorders, mild rash, muscle pain, pancreas inflammation, rapid heartbeat, skin inflammation or peeling, sleepiness, and slow heart beat The compositions are safe to use and are particularly advantageous due to the reduction or elimination of side effect that are associated with conventional anti-GERD drugs currently on the market. It is contemplated that the use of the plant extract of the present invention together with the traditional anti-GERD medicines will lower the dose of the traditional medicines that is required in order to reach a therapeutic effect, resulting in decreased side effects, which may result in increased patient compliance.
The Conyza bonariensis (L) Cronq extract and the at least one other agent can be formulated in the same composition, or they can be in different compositions.
If administered in separate compositions, the extract and the at least one other anti-GERD drug cam be administered sequentially or concurrently, in any order. For example, the extract can be administered followed by one or more conventional anti-GERD medicines.
Alternatively, conventional medicines can be administered prior to the extract.
Alternatively, the different components of the combination can be administered together, but in separate dosage forms.
Alternatively, the different compounds can be administered together in the same pharmaceutical composition. Further, if more than one of the aforementioned classes of compounds is administered, each component can be administered together or apart from the other. For example, if two or more conventional anti-GERD drugs are administered with the extract of the present invention, they can be administered in separate dosage forms, simultaneously or sequentially, in any order, or they can be provided in the same pharmaceutical composition, for concurrent administration.

Therapeutic Use and Mechanism of Action As contemplated herein, the present invention provides powerful methods for treating, preventing, reducing the severity of, or relieving the symptoms of GERD sucli as heartburn and regurgitation, and conditions associated with GERD, such as Esophagitis, NERD and Barrett's Esophagus.

As used herein, the term "heartburn" refers to a burning sensation in the chest and throat.

As used herein, the term "regurgitation" means a sensation of acid backed up in the esophagus.

Esophagitis: In most people, GERD symptoms are short-lived and occur infrequently.
In about 20% of cases, however, the condition becomes chronic. When the acid causes irritation or inflammation, the condition is called Esophagitis. If the damage becomes extensive and injures the esophagus, the disorder is known as Erosive Esophagus.
Non-Erosive Esophageal Reflux Disease: Symptoms of gastro esophageal reflux disease can occur without any signs of inflammation or injury to the esophagus. This condition is tentatively referred to as non-erosive esophageal reflux disease (NERD). NERD
rarely progresses to full-blown GERD. In NERD, patients have no signs of inflamination or erosion in the esophagus, but they experience certain symptoms of GERD, such as burning sensations behind the breastbone for at least three months. Research suggests that nerves lying near the surface of the lining become exposed to acid that has penetrated the layers. The nerves then trigger prolonged and painful symptoms in response.
Barrett's Esophagus: In a small percentage of chronic patients, a serious form of GERD called Barrett's esophagus may eventually develop, in which erosion can lead to cancerous changes in the tissue lining of the esophagus.
Though not wishing to be bound by any particular mechanism or theory, it is contemplated that the plant extract of the present invention is effective at treating the symptoms of GERD, by affecting the function of LES, so that food and digestive juices in the stomach cannot wash back into the esophagus. The extract further normalizes stomach and intestine functions, thereby leading to relief in symptoms such as bloating, spasmodic, vomiting, diarrhea and gas.
In one embodiment, the extract has anti-oxidant activity.
Pharmaceutical Compositions Although the active extract of the invention can be administered alone, it is contemplated that the extract will be administered in a pharmaceutical composition containing the active ingredient together with a pharmaceutically acceptable carrier or excipient.

Thus, in one einbodiment, the present invention provide pharmaceutical compositions useful for treating, preventing, reducing the severity of, or relieving the symptoms of GERD
and conditions associated with GERD, the compositions comprising a Conyza Less (e.g., a Conyza bonariensis (L) Cronq) extract, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD; and a pharmaceutically acceptable carrier or excipient.
Pharmaceutical compositions for use in accordance witli the present invention can be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations which can be used pharmaceutically. The active agents are formulated as pharmaceutical compositions and administered to a mammalian subject, such as a human patient in a variety of forms such as liquid, solid, and semisolid. The pharmaceutical compositions can be administered to a subject by any method known to a person skilled in the art, such as oral, topical, sublingual, buccal, parenteral, paracanceral, intravenous, transdermal, inhalation, intranasal, vaginal, intramuscular, transmucosal, transdermal, rectal, intradermal, subcutaneous, intraperitoneal, intraventricular, intracranial, rectal, vaginal, or intratumoral administration For oral administration, the compounds can be formulated by combining the active coinpounds with pharmaceutically acceptable carriers known in the art. The compositions can be forrnulated in any solid or liquid dosage form known in the art, including but not limited to, tablet, caplet, capsule (e.g., a soft or hard gelatin capsule), microcapsule, pellet, pill, powder, syrup, gel, slurry, granule, suspension, dispersion, emulsion, liquid, solution, dragee, bead and beadlet. The oral compositions can be formulated as iinmediate release formulations, or as controlled or sustained release formulations allowing for extended release of the active ingredient(s) over a predetermined time period.
Suitable excipients for solid formulations include but are not limited to fillers (diluents) such as sugars, including lactose, sucrose, mannitol, or sorbitol;
starch based excipients such as maize starch, wheat starch, rice starch, potato starch and the like, gelatin, gum tragacanth, phosphates such as calcium phosphate (e.g. calcium phosphate, dicalcium phosphate, tricalcium phosphate), cellulose based excipients as microcrystalline cellulose, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, methylhydroxypropylcellulose, hydroxypropylcellulose and the like. Polymers such as polyvinylpyrrolidone (PVP) and cross-lined PVP can also be used. In addition, the compositions may further comprise binders (e.g. acacia, cornstarch, gelatin, carbomer, ethyl cellulose, guar gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone), disintegrating agents (e.g. cornstarch, potato starch, alginic acid, silicon dioxide, croscarmelose sodium, crospovidone, guar gum, sodium starch glycolate), surfactants (e.g.

sodium lauryl sulfate), and lubricants (e.g. stearic acid, magnesium stearate, polyethylene glycol, sodium lauryl sulfate).
In another embodiment, the composition further comprises at least one vitamin.
Examples of suitable vitamins include, but are not limited to, vitamin A, vitamin B, vitamin C, vitamin D, vitamin E, and mixtures thereof.
For liquid formulations, pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Examples of oils include but are not limited to petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, and fish-liver oil.
Preferred oral pharmaceutical compositions include capsules made of gelatin as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Iii soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. In certain preferred embodiments the capsules exclude components of animal origin and are acceptable for vegetarians and vegans.
Soft gelatin capsules and methods of preparing them are known in the art. Non-limiting examples can be found in US Patent Nos. 6,217,902; 6,258,380;
5,916,591, and 4,891,229, all of which are incorporated herein by reference.
Other acceptable excipients and additives known to the person with skill in the art may be included in the compositions of the present invention, for example stabilizers, solubilizers, tonicity enhancing agents, buffer substances, preservatives, thickeners, anti-oxidants, complexing agents, flavor-improving agents such as sweeteners, e.g., aspartame, gellants, glidants, colorants and viscosity-increasing agents. A solubilizer can be for example, tyloxapol, fatty acid glycerol polyethylene glycol esters, fatty acid polyethylene glycol esters, polyethylene glycols, glycerol ethers or mixtures of those coinpounds. A
specific example of a solubilizer is a polyoxyethylated castor oil for example, the commercial products Cremophor or Cremophor RH40. Another example of a solubilizer is tyloxapol.
The concentration used depends especially on the concentration of the active ingredient. The amount added is typically sufficient to solubilize the active ingredient. For example, the concentration of the solubilizer is from 0.1 to 5000 times the concentration of the active ingredient.
Examples of buffer substances are acetate, ascorbate, borate, hydrogen carbonate/carbonate, citrate, gluconate, lactate, phosphate, propionate and TRIS
(tromethamine) buffers. The amount of buffer substance added is, for example, that necessary to ensure and maintain a physiologically tolerable pH range. The pH range is typically in the range of from 5 to 9, preferably from 5.2 to 8.5.
Tonicity enhancing agents are selected from ionic and non-ionic agents. For example, ionic compounds include alkali metal or alkaline earth metal halides, such as, for example, CaC12 KBr, KCI, LiCI, Nal, NaBr or NaCl, or boric acid. Non-ionic tonicity enhancing agents are, for example, urea, glycerol, sorbitol, mannitol, propylene glycol, or dextrose.
Examples of preservatives are quatemary ammonium salts such as benzalkonium chloride, benzoxonium chloride or polymeric quaternary ammonium salts, alkyl-mercury salts of thiosalicylic acid, such as, for example, thiomersal, phenylmercuric nitrate, phenylmercuric acetate or phenylmercuric borate, parabens, such as, for example, methylparaben or propylparaben, alcohols, such as, for example, chlorobutanol, benzyl alcohol or phenyl ethanol, guanidine derivatives, such as, for example, chlorohexidine or polyhexamethylene biguanide, or sorbic acid. Where appropriate, a sufficient amount of preservative is added to the composition to ensure protection against secondary contaminations during use caused by microbes. Where appropriate, anti-oxidants are also added.
The coinpositions of the present invention may comprise further non-toxic excipients, such as, for example, emulsifiers, wetting agents (surfactants) or fillers, such as, for example, the polyethylene glycols (PEG200, 300, 400 and 600) or Carbowax (Carbowaxl000, 1500, 4000, 6000 and 10000). Other excipients that may be used if desired are listed below but they are not intended to limit in any way the scope of the possible excipients.
They can be complexing agents, such as disodium-EDTA or EDTA, antioxidants, such as ascorbic acid, acetylcysteine, cysteine, sodium hydrogen sulfite, butyl-hydroxyanisole, butyl-hydroxytoluene; stabilizers, such thiourea, thiosorbitol, sodium dioctyl sulfosuccinate or monothioglycerol; or other excipients, such as, for example, lauric acid sorbitol ester, triethanol amine oleate or palmitic acid ester.

The amount and type of excipient added is in accordance with the particular requirements and is generally in the range of from approximately 1% to approximately 99%
by weight.
The amount of a composition to be administered will, of course, depend on many factors including the subject being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician. However, the dose employed will generally depend on a number of factors, including the age and sex of the patient, and the severity of the disease being treated.
Preferably, the preparations are in unit dosage form, intended for oral administration.
In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active components. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, for example, tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet itself or it can be the appropriate number of any of these in packaged form.
The following examples are presented in order to more fully illustrate certain embodiments of the invention. They should in no way, however, be construed as limiting the broad scope of the invention. One skilled in the art can readily devise many variations and modifications of the principles disclosed herein without departing from the scope of the invention.

EXPERIMENTAL DETAILS SECTION
Example 1: Oral Compositions A 375 capsule comprising a Conyza bonariensis (L) Cronq extract was prepared.
The capsule comprises 150 mg concentrated Conyza bonariensis (L) Cronq extract, 220 mg tricalcium phosphate (TCP), and 5 mg of vitamin E. The TCP was added as part of the drying process during extraction, as further explained in Example 5. This method of drying may be replaced by other methods such as spray drying.
In another embodiment, a 102 mg capsule comprising 50 mg Conyza bonariensis (L) Cronq extract, 50 mg TCP, and 2 mg vitamin E, was prepared.

Example 2: Toxicity:
To test the safety of the Conyza bonariensis (L) Cronq extract, an LDH release assay was performed. It was reported that LDH is released from the cells when plasma membrane integrity is destroyed by necrotic rupture.
The integrity of the plasma membrane was determined by measuring LDH activity released into the culture medium. LDH activity was monitored following the oxidation of NADH as the decrease in absorbance at 334 nm. The reaction was carried out in a potassium phosphate buffer (40 mM K2HPO4, 10 mM KH2PO4, pH 7.5), containing 0.24 mM NADH
and 0.62 mM pyruvate. The percentage of LDH released was defined as the ratio of LDH
activity in the supernatant to the sum of LDH amount released plus the activity measured in the cell lysate.
Human Fibroblast cells were incubated with different amounts of extract for 24h, 48h and 72h. In the end of the incubation time, the LDH activity was measured in the medium.
The results, depicted in Figure 1A-C, indicate that exposure of the cells to different amounts of the extract (180 g/ml and 360 g/ml) did not affect LDH release from the cells as compared to controls, after 24 hr (Fig 1A), 48 h (Fig 1B) or 72 h (Fig 1C).. The basal release of LDH from control cells was not statistically different from treated cells. This assures plasma membrane integrity during incubation of the cells with the extract.
The results show that cultured human fibroblast cells exposed to Conyza bonariensis (L) Cronq extract, did not exhibit any signs of toxicity nor was their growth diminished.
Example 3: Cytoprotective studies on animal and human cells Oxidative stress leads to generation of reactive oxygen species (ROS) which play an important pathogenic role in different disease-states. Lipid peroxidation has damaging effects on cell membranes. The extent of lipid peroxidation was measured using a technique based on a thiobarbituric acid reactive substance (TBARS) assay that detects malondialdehyde (1VIDA), an end product of peroxidative decomposition of polyeonic fatty acids in in-vitro systems. To accurately quantify TBARS in the analytical procedure, the protein was precipitated before the addition of thiobarbituric acid to the reaction, while the antioxidant butylated hydroxytoluene was added before heating of samples. Antioxidant properties of the product were investigated both in rat liver cells and in human sperm cells using the Bylond protocol for isolating the sperm membranes. Rat liver homogenates or human sperm cells were incubated with 100 M FeSO4 as ROS generating system and with various concentrations of the product.
Figure 2 shows the anti-oxidant effect of Conyza bonariensis (L) Cronq.
extract on 100 mM FeSO4 induced lipid peroxidation of rat liver homogenates. As can be seen, the extract has anti-oxidant properties, which increase as a function of the extract concentration.
Example 4: Mitochondrial Toxicity Testing (MTT) The mitochondrial toxicity effect of the Conyza bonariensis (L) Cronq extract was ,tested using a colorimetric assay based on the conversion of the yellow dye, tetrazolium bromide, to the red formazan derivative by mitochondrial succinate dehydrogenase in viable cells. Viable cells were obtained in a co-culture system of two cell lines, parenchymal and non-parenchymal cells. To evaluate hepatotoxicity, a co-culture of human hepatocytes (HepG2) and monocytes (THP1) was used. In this co-culture system both cell types have direct cell-to-cell contacts and are kept in more in-vivo culture conditions than in the monoculture system. The HepG2 cell line retains differentiated parenchymal functions of normal hepatocytes including the expression of P450isoenzymes, permitting long-term studies to be performed. To assess effects of the Conyza bonariensis (L) Cronq extract on cell viability both HepG2 and THP1 cell lines were used. The cell lines were grown in Dulbecco's modified Eagle's medium (DMEM), with a high glucose content (4.5 g/1) supplemented with 10% vol/vol inactivated fetal calf serum, with 1%
nonessential amino acids, l% glutamine, 100 units/ml penicillin and 10 mg/1 streptomycin. The cells were maintained in a humidified atmosphere with 5% CO2 at 37 C and the medium of these cells was changed twice a week. At 70-80% confluence, cells were trypsinized and seeded in 96 well plates in a cell density of 1.5 x 104 HepG2 cells and 5 x 103 THP1 cells.
After 24 hours of cell seeding, cells were incubated with various concentrations of the Conyza bonariensis (L) Cronq extract for another 24 hours at 37 C. Following removal of the extract from each well, cells were washed in phosphate-buffered saline. The cells were then incubated in serum-free DMEM to which the tetrazolium dye MTT (0.5 mg/ml) was added in each well (100 ml) and incubated for 4 hours. The medium was then removed and the cells were incubated for 15 min with 100 ml of acidic isopropanol (0.08 N HCl) in order to dissolve the formazan crystals. The absorbance of the MTT formazan was determined at 570 nm in an enzyme-linked immunosorbent assay (ELISA) reader. The cell viability was defined as the ratio of absorbance of treated cells to untreated cells and is expressed in the following as a percentage.
Figure 3 shows the results of an MTT assay in co-culture (HepG2, THP1) in control (untreated) cells and in cells treated with 10 g/ml and 100 g/ml Conyza bonariensis (L) Cronq extract. The results show that the plant extract did not exhibit any substantial signs of mitochondrial toxicity at the concentrations tested.

Example 4: Clinical Studies Patient Population In this study two groups were tested for the evaluation of the efficacy of the Conyza bonariensis (L) Cronq extract. The first group was previously treated with chemical anti-GERD medications, and the first group did not receive any previous treatment.
One hundred and four patients, ages 18-65, were treated with the 375 mg capsule described in Example 1.
Of them, fifty nine (59) had previously taken conventional drugs (all patients in this group had daily heartburn attacks), four (4) were pregnant women, and forty five (45) were not previously treated with heartburn medicines (patients in this group had weekly to monthly heartburn attacks). Patient demographics are summarized in Tables 1 and 2 below:

Table 1: Patient Distribution Age(y) BMI(kg/sq.m) Average 35.2 27.6 Stdev 5.42 3.58 Range 22-62 21-36 Table 2: GERD Parameters Average GERD
parameters Onset(y) 5 Severity 3.1 control 1.5 EES(%) 30 WBI 4.9 Methods Patients were given 1 capsule bid (375 mg) before meals for 10 days. Clinical follow-up was conducted for 4-6 weeks. The following parameters were evaluated:
1) Severity score 1-5.
2) Control of symptoms by current therapy (score 1-3).
3) Extra-esophageal symptoms recording (EES).
4) Gastrointestinal Symptom Rating Scale (GSRS) and the Psychological. General Well-Being (PGWB) index- combined as Integrated Well-Being. Index (WBI) (1-10 scale).
5) Relapse of symptoms.
The endpoints were:
1) Change of the severity of clinical symptoms.
2) Change in WBI (1-10 scale).
3) Change in EES.
4) Recurrence of symptoms within the follow-up period.
5) Medical therapy re-induction.
6) Adverse effects.
Efficacy:
Thirty five (35) Patients of the first group stopped suffering from heartburn 1-2 hours after the first capsule and stopped using conventional drugs, nineteen (19) of them reported that they felt heartburn after a week to 10 days and then took another capsule that stopped the heartburn for 7-10 days, sixteen (16) of them reported that the heartburn attacks stopped after 4-6 days of using our product tow capsules a day and then they did not feel heartburn for 5- 8 days (this group continued to use the product one capsule daily for tow weeks and reported that they did not have heartburn for 1-2 months). Eighteen (18) patients of the first group used the product (one capsule a day) for 15 days and reported that they started to feel better from the first day and the heartburn disappeared after 10-12 days for a minimum of 2 weeks, then the heartburn stopped after taking one or two capsules. Six (6) patients of the first group reported minor-no relief; and went back to conventional drugs. All pregnant women reported that the heartburn stopped after the first capsule for a minimum of one week.

The second group (45 patients) used one capsule when they had heartburn attack.
Thirty seven (37) patients reported that the heartburn disappeared after the first capsule for one month to two months. Four (4) patients reported that the product reduced the heartburn, and four (4) patients reported no effect.
The results are presented in Figure 4, and are quantified in Table 3 below:
Table 3 - Clinical Study Results After Before Severity 0.8 3.1 Control 2.7 1.5 EES 0% 30%
WBI 8 4.9 The results of this study show that treatment with Conyza bonariensis (L) Cronq extract is safe and effective alternative for treatment of GERD.

Example 5: Analytical Procedures The plant extracts were analyzed by HPLC and UV VIS spectroscopy .
Reagents : Acetonitrile : Water 70:30. All reagen.ts used - HPLC grade.

Sample Preparation: The plant was extracted in according to the block diagram depicted in Figure 5.

Extract with 99% Ethanol 15 kg. milled plant was extracted with 401cg ethyl alcoho199% for 2.5 hours at a teinperature of 70 C, then filtered through a fine filter. The residue was again extracted using another 40 kg ethyl alcoho199 % and filtered. The filtrates fiom both from extraction 1 and 2 extraction were mixed and evaporated.
69 kg of the filtrate at a starting total dissolved solids (TDS) (i.e., the solid materials inside the extract that include the active compounds) of 0.81 % was evaporated up to 30%
TDS at a teinperature less than 60 C with a final weight of 2.3 Kg.
The product can be supplied in a liquid foml or dried with a carrier (e.g., TCP). In this case the final prodtict consists of 40% active material and 60% inert carrier. ln the above extraction the product was divided into 1 kg 30 TDS and the rest absorbed and dried in a carrier resulting with 650 gram product consisting of 260 grams active material.

Extract with 50% Ethanol 15 kg. nlilled plant was extracted with 70 kg ethyl alcoho150% for 2.5 hours at a temperature of 70 C., and then filtered tlirough a fuie filter. The residue was again extracted using another 50 kg ethyl alcoho150 % and filtered. The filtrates from both from extraction 1 and 2 extraction were mixed and evaporated 105 kg of the filtrate at a stalting TDS of 1.7% was evaporated up to 30% TDS
at a temperature less than 60 C with a final weight of 5.95 kg.
The product can be supplied in a liquid form or dried with a carrier (e.g., TCP)... In this case the final product consists of 40% active material and 60% inert carrier. In the above extraction the product was divided into 1 lcg 30 TDS and the rest absorbed and dried in a carrier resulting with 3.6 lcg product consisting of 1.44 grams active material.

Extract with Water 15 kg. milled plant was extracted with 70 kg water for 2.5 hours at a temperature of 70 C., and then filtered through a fine filter. The residue was again extracted using another 50 lcg water and filtered. The filtrates from botli from extractions 1 and 2 were mixed and evaporated.

100 kg of the filtrate at a starting TDS of 2.1 % was evaporated up to 30% TDS
at a temperature less than 60 C with a final weight of 7 kg.

The product can be supplied in a liquid form or dried with a carrier (e.g., TCP). In this case the final product consists of 40% active inaterial and 60% inert carrier. In the above extraction the product was divided into 1 kg 30 TDS and the rest absorbed and dried in a carrier resulting with 4.3 kg product consisting of 1.7 kg active material.

Standard Preparation: Apigenin 0.7mg/ml - Fliilca 10798 Chromatographic Conditions HPLC type: HP 1090 Diode array Column: Phenomenex Luna 5u C18 Coluinn Teinperature: 40 C
Mobile Phase: isocratic Acetonitrile: Water 70:30 Flow Rate: 1.0 ml/min Detection: 270 nm and Diode array Inj ection Volume: 5 l.
Run Time: 20 minutes Pealcs Identification: Retention time and TJV/ VIS spectrum The Cliromatogram from the water extract 100% is shown in Figure 6A, and the UV
VIS spectruin of the main pealcs is shown in Figure 6B (retention time: 2.06 minutes).
The Chromatogram fiom the alcoholic extract 99% is shown in Figure 6C, and the UV VIS spectrums of the main peaks are shown in Figures 6D-F. Figtu=e 6D:
retention time:
2.29 minutes Figure 6E: retention tinle: 4.39 minutes. Figure 6F: rentention time:
4.74minutes The Chromatogram from the alcoholic extract 50% is shown in Figure 6G, and the UV VIS spectrunis of the main peaks are shown in Figures 6H-J. Figure 6H:
retention time:
2.29 minutes. Figure 6I: retention time: 4.45 minutes. Figure 6J: rentention time:
4.79minutes.
The Chromatogram of the standard Apigenun is shown in Figure 6K, and the UV
VIS
spectrum of the main peaks is shown in Figures 6L (retention time: 2.94 minutes).
The Yield from 100% water extract wasl4% based on TDS. The Yield from the 50%
alcoholic extract was 11.9% based on TDS. The Yield from the 99% alcoholic extract was 3.7% based on TDS. The peaks of the 50% and 99% alcoholic extracts were similar, and have lower intensities in coniparison to 100% water extract.
The total phenols were tested for each of the extracts 1. Alc 99% 0.05% in sol 2. Alc 50% 0.2% in sol 3. Water 0.14% in sol While certain embodiments of the invention have been illustrated and described, it will be clear that the invention is not limited to the embodiments described herein.
Nuinerous modifications, changes, variations, substitutions and equivalents will be apparent to those skilled in the art without departing from the spirit and scope of the present invention as described by the claims, which follow.

Claims (43)

1. A method of treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD) and conditions associated with GERD, comprising administering to a subject in need thereof a composition comprising an extract from a Conyza Less. plant, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD.

2. The method according to claim 1, wherein the extract is from a species of Conyza Less. selected from the group consisting of Conyza bonariensis (L) Cronq (asthma weed), Conyza canadensis (L.) Cronq. (Canadian horseweed); Conyza canadensis (L.) Cronq.
var. canadensis - (Canadian horseweed); Conyza canadensis (L.) Cronq. var. glabrata (Gray) Cronq. - (Canadian horseweed);
Conyza canadensis (L.) Cronq. var. pusilla (Nutt.) Cronq. -(Canadian horseweed); Conyza floribunda Kunth - (asthmaweed);
Conyza laevigata (L.C. Rich) Pruski - (manzanilla horseweed);
Conyza primulifolia (Lam.) Cuatrec. & Lourteig - (primroseleaf horseweed); and Conyza ramosissima Cronq (dwarf horseweed).

3. The method according to claim 1, wherein the extract is from Conyza bonariensis (L) Cronq.

4. The method of claim 1, wherein the symptoms of GERD are heartburn or regurgitation.

5. The method of claim 1, wherein the conditions associated with GERD are selected from esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.

6. The method of claim 1, wherein the composition comprises the extract in solid form.

7. The method of claim 1, wherein the composition comprises the extract in liquid form.

8. The method of claim 1, wherein the extract is a water extract.

9. The method of claim 1, wherein the extract is an alcoholic extract.

10. The method of claim 1, wherein the composition comprises from about 10% to about 90% by weight of the extract.

11. The method of claim 10, wherein the composition comprises about 40% to about 60% by weight of the extract.

12. The method of claim 1, wherein the extract comprises anti-oxidant activity.

13. The method of claim 1, wherein the composition further comprises at least one pharmaceutically acceptable carrier or excipient.

14. The method of claim 13, wherein the at least one pharmaceutically acceptable carrier or excipient is selected from the group consisting of fillers, diluents, binders, disintegrating agents, surfactants, lubricants, solvents, stabilizers, solubilizers, tonicity enhancing agents, buffers, preservatives, thickeners, complexing agents, emulsifiers, anti-oxidants, flavor-improving agents, gellants, glidants, sweeteners, colorants, viscosity-increasing agents, and any combinations thereof.

15. The method of claim 13, wherein the excipient is tricalcium phosphate.

16. The method of claim 1, wherein the composition comprises about 40% by weight of the extract, and about 60% by weight of tricalcium phosphate.

17. The method of claim 1, wherein the composition comprises about 50% by weight of the extract, and about 50% by weight of tricalcium phosphate.

18. The method according to claim 1, wherein the composition further comprises at least one vitamin.

19. The method according to claim 18, wherein the vitamin is vitamin E.

20. The method of claim 1, wherein the composition is administered via oral, topical, sublingual, buccal, parenteral, paracanceral, intravenous, transdermal, inhalation, intranasal, vaginal, intramuscular, transmucosal, transdermal, rectal, intradermal, subcutaneous, intraperitoneal, intraventricular, intracranial, rectal, vaginal, or intratumoral administration

21. The method according to claim 1, wherein the composition is administered orally.

22. The method according to claim 21, wherein the composition is in a form selected from the group consisting of a tablet, caplet , capsule, microcapsule, pellet, pill, powder, syrup, gel, slurry, granule, suspension, dispersion, emulsion, liquid, solution, dragee, bead and beadlet.

23. The method of claim 22, wherein the capsule is selected from a soft gelatin capsule and a hard gelatin capsule.

24. The method of claim 1, wherein the composition is administered once-daily, twice-daily, thrice daily, once-weekly or once-monthly.

25. The method of claim 1, wherein the composition is administered on an as-needed regimen so as to relieve the symptoms of GERD or conditions associated with GERD.

26. The method of claim 24, wherein the daily dose of the extract is from about 10 mg to about 2000 mg of extract.

27. The method of claim 1, wherein the composition is administered in combination with at least one other agent.

28. The method of claim 27, wherein the other agent is selected from the group consisting of antacids, H2-receptor agonists, proton-pump inhibitors, promotility agents, Sucralfate, and any combination thereof.

29. The method of claim 28, wherein the extract and the at least one other agent are included in the same composition.

30. The method of claim 28, wherein the extract and the at least one other agent are administered in separate compositions.

31. The method of claim 30, wherein the extract and the at least one other agent are administered sequentially.

32. The method of claim 30, wherein the extract and the at least one other agent are administered concurrently.

33. Use of an extract from a Conyza Less. plant, in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD).

34. Use according to claim 33, wherein the symptoms of GERD are heartburn or regurgitation.

35. Use of an extract from Conyza bonariensis (L) Cronq in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD).

36. Use according to claim 35, wherein the symptoms of GERD are heartburn or regurgitation.

37. Use of an extract from Conyza Less. in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.

38. Use of an extract from Conyza bonariensis (L) Cronq in the manufacture of a medicament for treating, preventing, reducing the severity of, or relieving the symptoms of, esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.

39. A method of treating, preventing, reducing the severity of, or relieving the symptoms of, Gastro Esophageal Reflux Disease (GERD) and conditions associated with GERD, comprising administering to a subject in need thereof a composition comprising an extract from Conyza bonariensis (L) Cronq, in an amount effective to treat, prevent, reduce the severity of, or relieve the symptoms of, GERD or conditions associated with GERD.

40. The method of claim 39, wherein the symptoms of GERD are heartburn or regurgitation.

41. The method of claim 39, wherein the conditions associated with GERD are selected from esophagitis, Non-Erosive Esophageal Reflux Disease (NERD), Barrett's Esophagus, and any combination thereof.

42. The method of claim 39, wherein the composition comprises about 40% by weight of the Conyza bonariensis (L) Cronq extract, and about 60% by weight of tricalcium phosphate.

43. The method of claim 39, wherein the composition comprises about 50% by weight of the Conyza bonariensis (L) Cronq extract, and about 50% by weight of tricalcium phosphate.