BOTANICAL EXTRACTS

FIELD OF THE INVENTION

[0001] The invention relates to the treatment of gastric ulcers and related symptoms in non-human animals, particularly equine animals, involving the use of compositions containing non-enteric forms of a proanthocyandin polymeric composition isolated from the plant Croton spp. or Calophyllum spp., or with an extract or latex derived therefrom. The compositions are useful in treating gastrointestinal ulcers in afflicted animals, thus providing a safe, effective and economical treatment for gastric or stomach ulcers, particularly in non-human animals of large size, such as horses.

BACKGROUND OF THE INVENTION

[0002] Ulcers, particularly gastrointestinal ulcers of the stomach and related conditions, can seriously affect the health and well-being of non-human animals, especially, large non- human animals, such as equine animals. Ulcers are lesions in the lining of the digestive tract and are very common in animals, particularly in horses that are used for activities involving endurance, competition and high-expectation performance.

[0003] Gastric ulcers, which occur in the stomach are common in large animals such as horses. Gastric ulcers in equines such as adult and young horses can be a source of great pain and distress for the afflicted animal. The symptoms of ulcers can manifest themselves in young (e.g., foals) and older animals in various ways, depending upon the animal and its experiences. For example, athletic and performance horses (hunter/jumpers, dressage, endurance and western) routinely develop ulcers. However, studies have shown that even small changes in the routine of a non-performance (recreational) horse can cause ulcers in as few as five days. Thus, horses do not have to undergo intensive exercise, activity, or stress to develop ulcers. Only a few days of activities typically associated with recreational riding has been found to cause ulcers in horses tested for ulcers after their involvement in such activities.

[0004] Gastric ulcers can affect horses at any age. Foals are often susceptible because they secrete gastric acid as early as two days of age and the gastric fluid is highly acidic. Foals that have infrequent or interrupted feeding or that are recumbant for long periods of time, have been found to have gastric fluid of a lower pH, suggesting that milk has a protective effect against stomach acid and that recumbancy increases the exposure of the stomach to acid. (U.C. Davis Center for Equine Health Horse Report, www.thehorse. com) .

[0005] Gastric ulcers are common in horses due to their anatomy and feeding conditions.

The stomach of a horse is smaller than the stomachs of other species. Because of this, horses cannot handle large amounts of food; instead, their stomach better supports grazing and ingestion of frequent, small portions of feed for extended periods of time. In a natural grazing situation, a horse requires a steady flow of acid for digestion; therefore, the stomach of a horse typically produces acid 24 hours a day, every day - up to 9 gallons of acidic fluid per day, even when the animal is not eating. In a natural, high-roughage diet, the acid produced is buffered by both feed and saliva. Id. However, when horses a fed on an "unnatural" schedule, such as only 2 times per day, for example, the stomach is subjected to a prolonged period in the absence of feed to neutralize the acid. In addition, high grain diets produce volatile fatty acids that also contribute to the development of ulcers.

[0006] Besides feeding and diet considerations, other risk factors for the development of gastric ulcers in non-human animals such as horses include physical and environmental factors, such as transport anxiety, stall confinement, intermittent feeding, lack of socialization. As determined by gastroscopy, a few hours of transport can induce gastric ulceration in horses than had no ulcers prior to departure. Moreover, chronic administration of non-steroidal antiinflammatory drugs (NSAIDs) can decrease the production of a horse's protective gastric mucous layer, leading to ulcer susceptibility in the animal. Id.

[0007] The equine stomach is divided into the squamous region at the top, which is considered to be a continuation of the esophageal lining, and the glandular mucosal region at the bottom. It is the glandular bottom mucosal region that secretes gastric acid, as well as produces mucous and bicarbonate, which protect the mucosa from acid exposure. Consequently, although the bottom gastric region is exposed to acid for several hours per day, it is less common for ulcer formation, given the buffering action of the mucosa. Ulcers in the stomach bottom are often caused by chronic administration of NSAIDs. The top area of the stomach is involved with mixing the contents of the stomach and has less protection from produced acid. Thus, gastric ulcers are often found here. The stomach lining in the top portion is thin and has fewer mechanisms for acid protection. Because the horse's stomach produces acid at all times, even when the animal is not eating, the squamous mucosa is exposed to acid several hours a day, which can readily erode the lining of the top stomach region.

[0008] Equine gastric ulcer syndrome (EGUS) refers to a spectrum of inflammatory and disruptive mucosal pathophysiology that affects tissues of the distal esophagus, stomach, and entrance into the duodenum. Endoscopic surveys indicate that approximately 80% of these lesions are found in the nonglandular squamous mucosa of the stomach, especially on the lesser curvature just proximal to the margo plicatus. However, significant portions of the squamous mucosa along the greater curvature and up into the fundus may also be involved, along with lesions in the antrum or pylorus.

[0009] Ulcers in the nonglandular squamous mucosa are associated with repeated direct insult from very low pH fluid normally found in the glandular region of the stomach. Pressure increases inside the animal's abdomen, which may be associated with exercise, and collapses the stomach, forcing the acid gastric contents upward. The increased and highly acidic fluid contents of the lower stomach come in contact with the nonglandular squamous mucosa, causing inflammation and potentially erosions to varying degrees.

[0010] The causes of ulcers in the glandular mucosa of the stomach are less well defined.

A possible cause related to the use of nonselective NSAIDs, which are known to reduce blood flow to the GI tract, which decreases production of the mucobicarbonate matrix by the gastric glandular mucosa, thus resulting in ulceration. While a correlation of the presence of Helicobacter spp organisms from the stomach of horses with and without gastritis and ulcers has been suggested, the results of relevant studies have not be definitive; therefore, the role of this organism in glandular equine gastric ulcers has not been determined.

[0011] Definitive diagnosis of ulcers in an animal requires minimally invasive gastric endoscopy or gastroscopy, which allows evaluation of the gastrointestinal system, including the esophagus, the squamous and glandular regions of the stomach and the proximal section of the small intestine in horses. Feed material can impede a complete endoscopic evaluation; therefore, an animal is usually fasted for a minimum of twelve hours prior to the procedure and examination.

[0012] Because the economic and humane impacts gastric ulcers, their symptoms and related conditions on afflicted animals are great, there is a significant need for new, medically effective and economical treatments and remedies for gastric ulcers, and their associated symptoms in non-human animals such as horses. The present invention addresses such needs by providing advantageous methods and compositions to treat and prevent gastric ulcers of the animal's digestive system.

SUMMARY OF THE INVENTION

[0013] The present invention provides a method of treating gastric (gastrointestinal) ulcers, including glandular and squamous ulcerations, and/or symptoms related thereto in a non- human animal, and treating EGUS, particularly, large animals such as horses, in which the method involves administering to an animal in need thereof a pharmaceutically acceptable composition that is not enterically protected or coated (herein termed "non-enteric"), comprising an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, formulated in effective amounts to treat the gastric ulcers and/or the symptoms thereof in the animal. The term "gastric ulcer" denotes a stomach ulcer and is used interchangeably with "gastrointestinal ulcer" herein.

[0014] It is an unexpected finding provided by the present invention that gastric ulcers may be treated, improved, or resolved in animals in need by administering a non-enteric formulation or composition comprising an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri (i.e., the non-enteric product) in the absence of an agent that inhibits or blocks acid production, such as an anti-acid (antacid), or a histamine type 2 (H-2) receptor antagonist which partially blocks acid production, or a proton pump (or proton-ion pump) blocking agent or inhibitor, such as omeprazole and the like. The present methods provide an advantage when compared with treatment involving commercially available anti-acid or proton pump inhibitors such as omeprazole. In particular, it has been surprisingly determined based on the studies described herein, e.g., Example 1, that treatment of EGUS with a formulation or composition comprising an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB-300, that is not enterically protected or coated, did not alter gastric pH of the animals during the treatment period, e.g., for 28 days of therapy with the non-enteric product, as well as for 7 days after treatment, relative to the gastric pH of animals prior to treatment (e.g., at the time of screening to determine baseline pH). Gastric pH during therapy with non-enterically coated or protected SB- 300 was observed to be similar to baseline gastric pH at all measured study time points. This finding stands in contrast to other ulcer treatments, e.g., proton pump inhibitors such as omeprazole, which act by blocking gastric acid secretion for the treatment and prevention of EGUS. By not altering the gastric pH while serving as an effective gastric ulcer treatment, the methods of the present invention, which comprise therapy with a non-enterically coated or protected formulation of an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB-300, are advantageous in the art. A treatment for EGUS that does not alter gastric pH in the animals undergoing treatment is important for several reasons, including maintenance of low gastric pH, which is essential for proper digestion; for gut immunity and as a first line defense against pathogens; for the absorption of vitamins and minerals; and for other potential downstream effects. A non-alteration of gastric pH in an animal undergoing treatment for gastric ulcers and EGUS with a non-enterically coated or protected formulation of an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB-300, refers to a statistically insignificant change in the gastric pH values measured in an animal during the course of the treatment compared with the gastric pH values measured in the animal at baseline or screening and/or compared with the gastric pH values of a placebo-treated animal. By way of nonlimiting example, a standard deviation between about +/-1.5 to +/-1.9 in variation of gastric pH values would be representative of a non-alteration in gastric pH between gastric pH measured in an animal at baseline or screening and gastric pH measured during the course of treatment of an animal, in particular, a horse.

[0015] In an embodiment, the methods of the invention treat, improve, or resolve glandular ulcerations in animals afflicted with gastric ulcers, particularly, horses. In an embodiment, the methods of the invention treat, improve, or resolve squamous ulcerations in animals afflicted with gastric ulcers, particularly, horses. In an embodiment, the methods of the invention treat, improve, or resolve glandular and squamous ulcerations in animals afflicted with gastric ulcers, particularly, horses. In an embodiment, the glandular and squamous ulcerations are improved or resolved in the treated animals, particularly horses, within about 2 weeks or by day 14 following treatment. In embodiments, the non-enteric composition comprising proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri (the product) is administered to treat gastric ulcers and associated ulcerations in a total daily dose of 200 mg to 50 g. It is envisioned that the total daily dose may be achieved by administering multiple daily doses of the non-enteric product, e.g., two, three, four or more doses per day, until the total daily dose is reached, as described further herein. In some embodiments, the non- enteric product is administered to an animal in need at a dose of 70-100 mg/kg/day. In other embodiments, the non-enteric product is administered to an animal in need at a dose of 10-15 mg/kg/day. In certain other embodiments, the non-enteric product is administered in a total daily dose of 500 mg or 250 mg twice daily (BID). In a particular embodiment, horses afflicted with glandular and squamous gastric ulcerations are administered 5 grams of a non-enteric composition comprising proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, in 2 doses per day, i.e., 2.5 g/dose/animal or 2.5 g BID. In a related embodiment, the non-enteric formulation of proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri is SB 300. In another particular embodiment, horses afflicted with glandular and squamous gastric ulcerations are administered 10 grams of a non- enteric composition comprising proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, in 4 doses per day, i.e., 10 g/dose/animal or 10 g BID. In a related embodiment, the non-enteric composition of proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri is SB 300. In accordance with the invention, no anti-acid, H-2 receptor antagonist, or proton pump inhibitor, such as omeprazole, is administered to the animal undergoing treatment. In addition, treatment of gastric ulcers (EGUS) with a non- enterically coated or protected composition of proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB 300, according to the present invention unexpectedly and advantageously does not alter the gastric pH of the animal (horse) undergoing treatment, thereby allowing for the maintenance of a low gastric pH, which significantly impacts the animal's digestive health and overall well-being as described herein. For example, a standard deviation between about +/-1.5 to +/-1.9 in variation of gastric pH values would be representative of a non-alteration in gastric pH between gastric pH measured in an animal at baseline or screening and gastric pH measured during the course of treatment of an animal, in particular, a horse.

[0016] In an aspect, the invention provides a method of preventing gastric ulcers in non- human animals, particularly equine animals such as horses. The non-enteric compositions of the invention can be administered to prevent first time ulcers or to prevent the recurrence of ulcers in the animals. As appreciated by those in the art, factors that may lead to gastric ulcers in horses, particularly performance equine athletes, include stall confinement, stress, intermittent feeding, intense exercise and administration of non-steroidal anti-inflammatories. The treatment and preventive methods of the invention may be used in conjunction with proper dietary and feeding oversight and management, as well as with methods of reducing and minimizing an animal's stress levels, for example, through better control and management of those aspects of an animal's environmental, lifestyle and living conditions that may lead to ulcerative conditions in the animal, for example, weaning, separation anxiety, housing, transportation, performance, endurance and competition. The methods of the invention aid in protecting, treating and improving the performance of equines afflicted with gastric ulcers and their symptoms.

[0017] The present invention preferably provides a method of treating and preventing gastrointestinal ulcers in equine animals, such as horses. The animals may be young, such as foals, or they may be adult, since both young and adult horses develop ulcers from various causes. In an aspect, the method is used to prevent, or prevent the recurrence of, gastric ulcers in a horse in need of treatment or prevention of a gastric ulceratic condition.

[0018] The methods of the invention also involve treating gastric ulcers as well as diarrhea, which frequently accompanies an ulceratic condition in a non-human animal, with a composition or formulation comprising an aqueous soluble proanthocyanidin from Croton lechleri or a Croton lechleri extract that is not enterically coated or protected. In an aspect, a pharmaceutically acceptable, non-enterically coated or protected composition comprising an aqueous soluble proanthocyanidin from Croton lechleri or a Croton lechleri extract is provided in a gel or paste formulation, which is orally administered. The methods of the invention do not include an anti-acid or proton pump inhibitor or blocker, such as omeprazole and the like. In addition, the methods of the invention do not alter the gastric pH of the animal undergoing treatment, thereby maintaining a low gastric pH during treatment with the non-enterically coated or protected composition comprising an aqueous soluble proanthocyanidin from Croton lechleri or a Croton lechleri extract, which promotes a more normal gastric environment during, as well as subsequent to, treatment.

[0019] In embodiments of the above methods of the present invention, an extract from

Croton lechleri is included in a non-enteric formulation or composition. In other embodiments, a pharmaceutically acceptable C lechleri proanthocyanidin polymer is included in a non-enteric formulation or composition. In embodiments of the methods, the C. lechleri proanthocyanidin polymer or C. lechleri botanical extract in the non-enteric composition is SB 300, SP 303, or crofelemer.

[0020] In embodiments, the non-human animal is a young or juvenile animal, or an adult animal. The method of the invention is not particularly limited as to the species of the animal and may include exotic, zoo, farm, or domestic animals, e.g., without limitation, equine, ovine, swine animals, goats, bison, buffalo, or camels, especially if such animal species are adversely affected by gastric ulcers and related conditions. In particular or preferred embodiments, the non-human animal is an equine animal (an adult horse or a young horse (foal)). In an embodiment, the non-human young or adult animal afflicted with gastric ulcers is additionally suffering from diarrhea, e.g., diarrhea associated with the gastric ulcer condition.

[0021] In particular embodiments, a non-enteric composition comprising the C. lechleri proanthocyanidin polymer or botanical extract is orally administered to an animal, e.g., an adult horse, in an dosage amount of 250 mg twice per day (BID), or in a dosage amount of 2.5 g twice per day (BID), (a total of 5 grams per animal per day), or in a dosage amount of 10 g four times per day (QID), (a total of 40 grams per animal per day). In an embodiment, the C lechleri proanthocyanidin polymer or botanical extract in the non-enteric composition or formulation is SB 300, SP 303, or crofelemer. In an embodiment, the C. lechleri proanthocyanidin polymer or botanical extract in the non-enteric composition is SB 300.

[0022] In a particular embodiment, the non-enteric composition comprising the C lechleri proanthocyanidin polymer or botanical extract is in the form of a paste, gel, or gel paste suitable for oral administration and for delivery of the active, C. lechleri proanthocyanidin polymer or botanical extract throughout the digestive tract, including to the stomach. Such pastes, gels, or gel pastes are typically and advantageously administered to the animal by topical application to the roof of the animal's mouth. In an embodiment, the non-enteric composition comprising a C. lechleri extract or C. lechleri proanthocyanidin polymer composition is in the form of a gel, paste, or gel paste formulation for oral administration and delivery to the stomach of an animal in need. In some cases, the gel, paste, or gel paste is contained in a delivery device, which can be a syringe, such as a needle-less syringe. In embodiments of the methods, the C. lechleri extract or the C. lechleri proanthocyanidin polymer are formulated in compositions, which are not enterically coated or protected, in amounts effective to reduce, decrease, diminish, inhibit, attenuate, or block acid production and to thereby treat, reduce, ameliorate, or prevent gastrointestinal ulcers or stomach ulcer production in the animal to which the composition is administered. In embodiments of the methods of the present invention, the C. lechleri proanthocyanidin polymer or C. lechleri botanical extract in the non-enteric composition is SB 300, SP 303, or crofelemer. In an embodiment, the methods of the present invention involving treatment of EGUS with a non-enterically coated or protected formulation or composition comprising a C. lechleri proanthocyanidin polymer or C. lechleri botanical extract, e.g., SB 300, SP 303, or crofelemer, does not alter and essentially maintains the gastric pH of the animal relative to the gastric pH of animals prior to treatment (e.g., at the time of screening and the determination of baseline gastric pH value), within normal variation, during the treatment period, as well as following the treatment period. In a preferred embodiment, the method involves treatment with SB 300.

[0023] In an aspect, the invention provides a method of treating or preventing gastric ulcers in a non-human animal, particularly an adult or young horse, that has, or that is at risk of developing, stomach ulcers, comprising administering to the animal in need thereof an effective amount of the above-described non-enterically protected composition of the invention. In an embodiment, the composition is orally administered. In an embodiment, the non-enterically protected composition is administered to a young or adult animal, in particular, a horse, that has, or is suspected of having, a gastric ulcer, and/or symptoms thereof, or that is at risk of developing a gastric ulcer and/or symptoms thereof. In some embodiments, the non-enteric products and compositions of the invention may be co-administered with other drugs or agents, for example, probiotics, which may serve to increase the efficiency of bacterial enzyme degradation. In other embodiments, the methods of the invention do not include an anti-acid or proton pump inhibitor or blocker, such as omeprazole and the like.

[0024] In embodiments, the non-enterically protected compositions and formulations comprising a C. lechleri proanthocyanidin polymer or a C. lechleri botanical extract employed in the methods are provided to an animal in need over a predetermined time period, i.e., for chronic administration. In other embodiments, the non-enteric compositions and formulations of the methods are provided to an animal in need over a discrete period of time, i.e., acute administration, until the gastric ulcers or related symptoms have abated, are reduced and/or improved, or are eradicated.

DETAILED DESCRIPTION OF THE INVENTION

[0025] Ulceratic lesions in the mucosal lining of the stomach and digestive tract are very common in equine animals, e.g., horses, used for recreation, commercial, performance and agricultural purposes. In particular, many competitive activities, including racing, dressage, show jumping, endurance events and western performance, lead to stomach ulcers and related symptoms in horses participating in these activities. It has been found that, depending on the intensity of training, the prevalence of gastric ulcers in horses can vary from 10 to 90%. Apart from exercise, many factors contribute to the development of gastric ulcers, for example, transport to and from show grounds, stall confinement in unfamiliar surroundings, stress and/or irregular feeding schedules.

[0026] Equine animals such as horses may suffer from gastrointestinal ulcers in the stomach. About 87% of horses involved in competitive activities and about 37% of horses not involved in competition, i.e., leisure or recreational horses, have gastric ulcers. The present invention provides compositions and formulations that are non-enterically coated or protected (i.e., non-enteric) comprising a C. lechleri proanthocyanidin polymer or a C. lechleri botanical extract for use in methods that can conveniently, efficiently and economically treat, improve and resolve stomach ulcers in animals in need, especially adult and young horses. In embodiments, the C. lechleri proanthocyanidin polymer or C. lechleri botanical extract in the non-enteric composition is SB 300, SP 303, or crofelemer. The methods of the invention do not include proton pump inhibitors such as omeprazole, or other anti-acids or H-2 receptor inhibitors.

[0027] There is a need in the veterinary field for new and improved medications and treatments for gastric ulcers in recreational and performance animals such as horses. The digestive tracts of large animals such as horses are unique and provide challenges to administering drugs whose activities are required in the stomach and digestive tract for effective treatments. For example, the digestive physiology of the horse is characterized by rapid gastric transit, rapid but intense enzymatic activity along the small intestine, and a long and intense microbial fermentation in the large intestine. The present non-enterically coated compositions and methods are particularly suitable for fulfilling the need of new and effective therapies and delivery approaches for treating gastrointestinal ulcers in non-human animals, such as horses.

[0028] The symptoms of ulcers in affected animals can be subtle and may include irritability and changes in attitude and behavior (for the worse), poor appetite and decreased performance and energy. These symptoms may also be exhibited with other symptoms such as lethargy; musculo/skeletal discomfort and pain; decline or deterioration in body condition and/or appearance (dull hair coat); weight loss; alterations in eating or drinking patterns; resistance to grooming; reluctance or refusal to performing certain tasks; and behavior indicating discomfort, such as pawing or laying down excessively. Still other signs that may correlate with ulcers in the animal include sensitivity in the flank area and girthiness; cribbing (windsucking); wood- chewing; and weaving in the stall, an (atypical) unwillingness to work or cooperate, and resistance under saddle. Foals afflicted with a gastric ulcer may also grind their teeth or lay on their backs. Once an ulcer is suspected or determined in a non-human animal such as horses, ponies, camels, donkeys, or mules, particularly, horses, the methods of the invention are effective in treating, improving, and resolving gastric ulcers, including glandular and/or squamous ulcerations, and/or the symptoms of such ulcers and ulcerations in the animal, for example, as described in Example 1 herein.

[0029] The present invention provides methods directed to treating stomach ulcers and the uncomfortable and often painful discomfort that they cause in both young and adult horses and other non-human animals. The methods are effective in reducing, improving and/or alleviating gastric ulcers and the symptoms that afflict such animals in need thereof. Treatment and prevention of gastric ulceratic conditions in horses with the methods and compositions of the invention can positively impact and improve the performance of horses that are expected to perform at peak proficiency, including leisure and recreational horses and show horses, especially training and race horses. The invention further provides non-enterically coated formulations and compositions comprising a C. lechleri proanthocyanidin polymer or C. lechleri botanical extract suitable for treating and preventing ulcers in young and adult animals. Unless otherwise noted herein, use of the term "animal" herein denotes non-human, warm-blooded mammals of a number of different species. Without wishing to be limiting, "young" or "juvenile" animals are generally about one year of age or under one to two years of age.

[0030] It is also to be understood that, even if not specifically stated herein, the methods and non-enteric compositions of the invention are intended for the therapy and the prevention of gastric ulcers that afflict both young and adult animals, such as horses, that are prone or susceptible to, or are at risk of, developing gastric ulcers, depending on their living situations, activities, or experiences. Preventative or proactive administration of the non-enteric compositions and formulations according to the methods of the invention can be provided to an animal prior to, or around the time of, stressful events, performances, transport, or relocation, for example. Animals (horses) having a history of gastric ulcers may benefit from proactive treatment with the compositions of the invention to decrease or abrogate the chances of developing gastric ulcers or the recurrence of ulceration, including glandular and/or squamous ulceration. During and following a course of treatment with the non-enterically coated or protected compositions and methods described herein, an animal being treated can be monitored for a change in its clinical behavior to determine that the gastric ulceratic condition has improved, or is reduced or eliminated. Preferably, the animal is examined via endoscopy or gastroscopy to confirm improvement and/or healing of ulceratic lesions, and prior to discontinuing therapy with the methods and compositions of the invention. Endoscopic examination involves short-term tranquilization of the animal to reduce stress to the animal from the procedure. Thereafter, an endoscope is inserted through the animal's nostril and guided down the esophagus into the stomach wherein the light and camera on the endoscope's terminus allow observation of the stomach lining. A complete endoscopic evaluation can take about 10 to 20 minutes and is safe for the animal. [0031] The present invention embraces therapeutic and preventative non-enterically coated or protected compositions and formulations to treat and avert gastric ulcers in regions of the digestive tract, particularly, the stomach, and related symptoms in animals, especially horses, experiencing digestive discomfort, as well as in animals at risk of or prone to digestive or gastrointestinal problems or ulcers. Signs that can indicate a digestive problem such as gastric ulcers in an animal such as a young or adult horse include the following: weight loss and/or general decline in body condition; resistance under saddle; irritability and other changes in attitude; lack of energy and stamina; loss of appetite; behavior indicating discomfort, such as pawing or laying down excessively; and low-grade anemia. The non-enteric compositions and methods described herein may be administered to an animal or horse that exhibits any of the foregoing symptoms and/or if appropriate testing indicates that the animal or horse is likely to be suffering from gastric ulcers. The methods and non-enterically coated or protected compositions of the invention provide a solution to a significant need for the animal industry, particularly for performance animals used for sport and competitive activities, for treating and preventing gastric ulcers which may be debilitating in afflicted animals. In addition, the methods of the invention comprising the non-enterically coated or protected C. lechleri proanthocyanidin polymer or C. lechleri botanical extract do not include proton pump inhibitors such as omeprazole, or other proton pump inhibitors or partial antagonists, or antacids. In an embodiment, the C. lechleri botanical extract SB 300 in a non-enterically coated or protected formulation or composition is used in the methods of the invention.

[0032] As appreciated by the skilled practitioner in the art, the horse stomach continuously secretes variable amounts of hydrochloric acid throughout the day and night, and secretion of acid occurs without the presence of feed material. Foals secrete gastric acid as early as 2 days of age and acidity of the gastric fluid is high. High acid in the stomach may predispose foals to EGUS. In adult horses, the stomach secretes approximately 1.5 liters of gastric juice per hour, and acid output ranges from 4 to 60 mmoles of hydrochloric acid per hour. The pH of a horse's gastric contents ranges from pH 1.5 to 7.0 depending on the region measured. A near neutral pH can be found in the dorsal portion of the esophageal region of the stomach (saccus cecus) near the lower esophageal sphincter. More acidic pHs are found near the margo plicatus (pH 3.0 to 6.0) and in the glandular region near the pylorus (pH 1.5 to 4.0). Gastric emptying of a liquid meal occurs within 30 minutes, whereas complete gastric emptying of a roughage hay meal typically occurs in 24 hours.

[0033] As noted above, the present methods offer unique advantages to the treatment of gastric ulcers and EGUS in animals in need, such as horses. Based on an analysis of the study results described herein, the treatment of EGUS with a non-enterically coated or protected formulation or composition comprising an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB-300, did not alter gastric pH of the animals during the treatment period, e.g., for 28 days of therapy with the non-enteric product, as well as for 7 days after treatment. The present methods do not significantly modify or change the pH of the animal's gastric regions tested, compared with the pH values as determined in the gastric regions prior to the treatment methods, i.e., baseline gastric pH, such as the gastric pH ranges mentioned above. The gastric pH during therapy with the non-enterically coated or protected SB-300 was observed to be similar to baseline gastric pH at all of the time points measured during the study described in Example 1. Accordingly, gastric pH values in the animals tested during treatment with a non-enterically coated or protected formulation or composition comprising an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB-300, (e.g., a glandular region pH value of 1.5 to 4.0), did not fluctuate significantly and were not significantly different from the gastric pH values determined at baseline or screening and prior to treatment, or from the gastric pH values of placebo-treated animals. As mentioned, a standard deviation between about +/-1.5 to +/-1.9 in gastric pH values as measured in animals undergoing the treatment methods of the present invention compared with baseline gastric pH measurements of the animals and/or gastric pH measurements of placebo controls would be considered as not altering gastric pH.

[0034] Thus, the treatment and therapy methods of the present invention are unlike other or commonly-used ulcer treatments, e.g., proton pump inhibitors, such as omeprazole, that act by blocking gastric acid section for the treatment and prevention of EGUS in animals such as horses. By not altering or significantly changing the gastric pH of a treated animal, while serving as an effective gastric ulcer treatment, the methods of the present invention, which comprise therapy with a non-enterically coated or protected formulation of an aqueous soluble proanthocyanidin from Croton lechleri, or an extract or latex derived from Croton lechleri, such as SB-300, are distinct from other methods and are beneficial to the animals undergoing ulcer therapy. The present treatment therapy for EGUS that does not alter gastric pH in treated animals is important for several reasons, including maintenance of low gastric pH in the animal, which is essential for proper digestion; for gut immunity and as a first line defense against pathogens; for the absorption of vitamins and minerals; and for other potential downstream effects.

[0035] The methods, non-enteric compositions and treatments of the invention are particularly suitable for treating animals of a young age, as well as adult animals, that suffer from, or are vulnerable to developing ulcers of the digestive tract, such as gastric ulcers. In an embodiment, the animals are young or juvenile non-adult animals that are born, bred, raised and/or maintained in a domesticated, performance, event and/or agricultural setting, e.g., animals raised and maintained for commodities such as labor, sport, performance, endurance, or other commercial or non-commercial capacity. In an embodiment, the animals are adult animals. Nonlimiting examples of animals affected by stomach ulcers and treatable by the methods and non-enteric formulations of the invention particularly include young (foals) and adult equine animals (horses). Other animals that may suffer from gastric ulcers and benefit from treatment and prevention by the methods and compositions described herein include young camels (calves) and adult camels. In addition, young cattle (calves), pigs (piglets), sheep (lambs), goats (kids), horses (foals) and adult animals, including, cattle, steer, bison, buffalo, goats, sheep and rams, may be treated according to the methods of the invention. In particular embodiments, both young and adult animals may be administered the non-enterically coated compositions comprising a Croton spp-derived proanthocyanidin polymer or botanical extract to prevent and treat stomach ulcers and related conditions in an animal or population of animals having or suspected of having such ulcers, for example, based on diagnostic testing and/or observations of behavior and symptoms indicative of gastric ulceratic conditions as described herein.

[0036] In an embodiment, a gel or paste formulation product is employed to orally administer to non-human animals and deliver to the digestive tract of an animal a C. lechleri proanthocyanidin polymer or C. lechleri extract in a non-enteric composition or formulation, such that the C. lechleri proanthocyanidin polymer or C. lechleri extract will be active and effective for treating, improving, resolving, preventing, or reducing the risk of gastric ulcers, ulcer formation, and related symptoms and conditions in an animal in need. The preparation is particularly suited for treating horses. In embodiments, the not enterically coated or protected compositions or formulations comprise a C. lechleri proanthocyanidin polymer or C. lechleri botanical extract that is SB 300, SP 303, or crofelemer. In a particular embodiment, C. lechleri proanthocyanidin polymer or C. lechleri botanical extract is SB 300.

[0037] In general terms, "treating" an animal according to the present methods refers to achieving or obtaining a desired physiologic and/or pharmacologic effect, whether prophylactic, therapeutic, or both. As used herein "treating" or "treatment" can refer to ameliorating, preventing, inhibiting, reversing, attenuating, alleviating, abrogating, minimizing, suppressing, reducing, resolving, decreasing, diminishing, stabilizing, eradicating, curing, or eliminating the deleterious effects of a disease or condition, or the progression or worsening of the disease or condition. For example, successful treatment may involve improving, resolving, or alleviating one or more symptoms of a disease or condition, although not necessarily all of the symptoms, of the disease or condition, or attenuating the symptoms or progression of the disease or condition. Curing or eliminating the disease or condition from the animal is an optimal outcome of the practice of the methods of the invention.

[0038] According to the invention, treatment of an animal in need thereof typically involves the use or administration of effective amounts or therapeutically effective amounts of a proanthocyanidin polymer or a proanthocyanidin polymer, or an extract or sap, preferably from a Croton spp. particularly C. lechleri, formulated in a non-enterically protected composition. For use in the stomach ulcer treatment methods, the C. lechleri proanthocyanidin polymer or extract is a non-enterically protected product, without a proton pump inhibitor or blocker, such as omeprazole or other proton pump inhibitors. Effective amount refers to the quantity (amount) of the drug and/or the composition containing the drug and the like, that induces a desired response in the animal subject upon administration or delivery to the animal. Optimally, an effective amount produces a therapeutic effect in the absence of, or with little or virtually no, adverse effects or cytotoxicity in the animal. Alternatively, any adverse effects associated with an effective amount are optimally outweighed by the therapeutic benefit achieved.

[0039] The treatment methods are directed to ameliorating, preventing, inhibiting, reversing, attenuating, alleviating, abrogating, minimizing, suppressing, reducing, decreasing, diminishing, stabilizing, resolving, improving, eradicating, curing, or eliminating gastric ulcers and/or their associated symptoms that adversely affect the health, lifestyle and performance of young and adult animals, such as horses. In an embodiment, the gastric ulcer is a stomach ulcer which is treated, improved, or resolved in an animal. In an embodiment, glandular ulcerations in the animal are treated, improved, or resolved by the methods of the invention. In an embodiment, squamous ulcerations in the animal are treated, improved, or resolved by the methods of the invention. In an embodiment, glandular and squamous ulcers are simultaneously treated, improved, or resolved by the methods of the invention. In other embodiments, the methods and non-enterically coated compositions of the invention also and advantageously improve and treat diarrhea, e.g., secretory/watery diarrhea, that may be associated with the gastric ulcers and ulceratic conditions in an afflicted animal.

[0040] In general, due to the anatomy of the gastrointestinal tract of adult animals such as horses, ulceratic conditions affecting the stomach may also cause diarrhea. Young animals, e.g., foals, that are less than about three months of age do not have fully competent gastric systems as do adult animals; therefore, young animals tend to be more prone to diarrhea caused by ulceratic conditions. In general terms, a foal is an equine, particularly a horse, that is one year old or younger in age. In accordance with the invention, the described compositions and methods can be administered to an animal suffering from gastric ulcers and/or related symptoms and also experiencing associated diarrhea so as to treat both the gastric ulcers and the associated diarrhea.

Proanthocyanidins and Tannins Obtained from Plant Extracts

[0041] Proanthocyanidins are types of condensed tannins, which are found in a large number of plants and are classified as hydrolyzable or condensed. Tannins and, in particular, proanthocyanidins are contained in many plants used in traditional medicine as treatment or prophylaxis for diarrhea {See, e.g., Yoshida et al., 1993, Phytochemistry, 32: 1033; Yoshida et al., 1992, Chem. Pharm. Bull., 40: 1997; Tamaka et al., 1992, Chem. Pharm. Bull, 40:2092).

[0042] Proanthocyanidins are comprised of at least two or more monomer units that may be of the same or different monomelic structure. The monomer units (generally termed "leucoanthocyanidins") are generally monomelic flavonoids which include catechins, epicatechins, gallocatechins, galloepicatechins, flavanols, flavonols, flavan-3,4-diols, leucocyanidins and anthocyanidins. The polymer chains are thus based on different structural units, creating a wide variation of polymeric proanthocyanidins and a large number of possible isomers (Hemingway et al., 1982, J. C. S. Perkin, 1 : 1217). Larger polymers of the flavonoid 3- ol units are predominant in most plants and often have average molecular weights above 2,000 daltons (Da), containing 6 or more units (Newman et al., 1987, Mag. Res. Chem., 25: 118).

[0043] Proanthocyanidin polymers and proanthocyanidin are found in a wide variety of plants, especially those having a woody habit of growth (e.g., Croton spp.. and Calophyllum spp.). A number of different Croton tree species, including Croton sakutaris, Croton gossypifolius, Croton palanostima, Croton lechleri, Croton erythrochilus and Croton draconoides, which are endemic to South America, produce a red viscous latex sap called Sangre de Drago (also known as Sangre de Grado) or "Dragon's Blood". The red viscous latex sap is known for its medicinal properties. For example, U.S. Patent No. 5,211,944 describes the isolation of an aqueous soluble proanthocyanidin polymer composition from Croton spp. (See also, Ubillas et al., 1994, Phytomedicine, 1 :77). The isolation of an aqueous soluble proanthocyanidin polymer composition from Calophyllum inophylum in U.S. Patent No. 5,211,944.

[0044] In an embodiment, an extract (botanical extract) from C. lechleri, such as the viscous sap which is extracted from the C. lechleri tree or the herbal medicine, Sangre de Drago, may be used in the compositions described herein. Sangre de Drago obtained from Croton trees is available throughout the Amazon, especially in the upper jungle of Peru and Ecuador, where such trees grow rapidly and can reach heights of 30-45 feet in three years. The sap (or extract) can be harvested, derived, or obtained from the Croton trees, in a manner akin to that of obtaining rubber or maple syrup. More ecologically-minded farming techniques are used to improve the cycles in which trees from which the sap is derived are grown, tapped and felled for sustainability. As used herein, a C. lechleri extract, or botanical extract, can also refer to C. lechleri sap (e.g., the latex sap or viscous sap) extracted from the C. lechleri tree.

[0045] In an embodiment, a proanthocyanidin polymer from C. lechleri, or a non- enterically coated or protected composition thereof, for use in the compositions and methods of the present invention is crofelemer. Crofelemer (CAS 148465-45-6) is an oligomeric proanthocyanidin of varying chain lengths derived from the Dragon's Blood of Croton lechleri, a tree of the family Euphorbiaceae, which is sustainably harvested under fair trade work practices in the Amazon. It has an average molecular weight of approximately 1900 Da to approximately 2700 Da. The monomers comprising crofelemer comprise catechin, epicatechin, gallocatechin, and epigallocatechin. The chain length of crofelemer ranges from about 3 to about 30 units with an average chain length of about 8 units. Crofelemer has the chemical formula: (Ci506 _,7 Hi ₂ ) _n and a molecular mass of 860-9100 g/mol. The antisecretory mechanism of action of crofelemer involves the targeting and inhibition of two, distinct intestinal chloride channels, namely, the cystic fibrosis transmembrane regulator conductance (CFTR) channel, which is a cAMP- stimulated CI ^" channel, and the calcium-activated chloride channel (CaCC), as reported, for example, by Tradtrantip, L. et al., 2010, "Crofelemer, an Antisecretory Antidiarrheal Proanthocyanidin Oligomer Extracted from Croton lechleri, Targets Two Distinct Intestinal Chloride Channels", Mol. Pharmacol., 77(l):69-78). A general structure of crofelemer is shown below. In the structure, an H at the R position of the structure signifies procyanidin; an OH at the R position of the structure signifies prodelphinidin.

[0046] In accordance with an embodiment of the invention, a non-enterically coated or protected formulation of crofelemer, or a pharmaceutically acceptable non-enteric formulation or composition comprising crofelemer, is employed in the treatment methods and compositions as the proanthocyanidin polymer from Croton lechleri.

[0047] In an embodiment, a non-enterically coated or protected formulation of SP 303, an oligomeric proanthocyanidin from Croton lechleri, (also known as crofelemer) is the proanthocyanidin polymer from Croton lechleri, or a pharmaceutically acceptable non-enteric formulation or composition comprising SP 303, is suitable for use in the treatment methods of the invention. SP-303 (R. Ubillas et al., 1994, Phytomedicine, 1 :77-106) is largely composed of purified proanthocyanidin oligomers (-)-galloepicatechin and (+)-gallocatechin,(-)-epicatechin and (+)-catechin and is suitable for use in the non-enterically coated or protected formulations and compositions for administration in the treatment methods described herein.

[0048] In another embodiment, a non-enterically coated or protected formulation of SB

300, a proanthocyanidin polymer extract from Croton lechleri is the proanthocyanidin polymer from Croton lechleri, or a pharmaceutically acceptable non-enteric formulation or composition comprising SB 300, is suitable for use in the treatment methods of the invention. SB 300, as described, for example, by Fischer, H. et al., (2004, J. Ethnopharmacol, 93(2-3):351-357) provides a natural product extract that is particularly amenable for use the non-enterically coated or protected formulations and compositions, and is highly functional and cost-effective in the treatment methods described herein.

[0049] In an embodiment, a pharmaceutically acceptable composition comprising a proanthocyanidin polymer from Croton lechleri and employed in the treatment methods of the invention can be obtained from C lechleri, e.g., as described in WO 00/47062 to Shaman Pharmaceuticals, Inc., the contents of which are incorporated herein, and formulated as a food or dietary supplement or nutraceutical formulation, especially in a non-enterically coated formulation.

[0050] In other embodiments as noted above, the non-enteric compositions useful in the methods of the invention comprise a raw latex obtained from a Croton species or a Calophyllum species, or an extract (botanical extract) or sap obtained from a Croton species or a Calophyllum species, which are not specifically polymeric proanthocyanidin polymer compositions. Exemplary extracts are described in Persinos et al., 1979, J. Pharma. Sci., 68: 124 and Sethi, 1977, Canadian J. Pharm. Sci., 12:7 ^' .

[0051] In accordance with the invention, the Croton spp-derived proanthocyanidin polymer or extract is formulated without an enteric coating or matrix. In embodiments, In particular or preferably, a non-enterically coated or protect formulation or composition comprising the proanthocyanidin polymer from Croton lechleri, or Croton lechleri extract, for example, SB 300, is intended for use in the methods of the present invention.

Preparation of Proanthocyanidin Polymer Compositions and Formulations

[0052] The proanthocyanidin polymer composition, effective for treating stomach ulcers according to the invention, is comprised of monomeric units of leucoanthocyanidins. More particularly, the composition is comprised of proanthocyanidin polymers of 2 to 30 flavonoid units, preferably 2 to 15 flavonoid units, more preferably 2 to 11 flavonoid units and most preferably an average of 7 to 8 flavonoid units with a number average molecular weight of approximately 2500 Da. The proanthocyanidin polymer composition is preferably soluble in an aqueous solution. In embodiments, for use in the methods according to the invention is a non- enterically coated formulation comprising a proanthocyanidin polymer from C lechleri; such a C lechleri proanthocyanidin polymer may in some cases be in the form of a pharmaceutically acceptable non-enteric product.

[0053] Examples of proanthocyanidin polymeric compositions useful in the present invention are preferably isolated or purified from a Croton spp., namely, Croton lechleri, or Calophyllum spp. by any method known in the art. For example, the proanthocyanidin polymer composition may be isolated from a Croton spp. or Calophylum spp. by the method disclosed in U.S. Pat. No. 5,211,944 or in Ubillas et al. (1994, Phytomedicine, 1 :77-106, called SP 303 therein), both of which are incorporated herein by reference. Other isolation methods are described in U.S. Patent Nos. 7,556,831 and 8,067,041 (Example 2), the contents of which are incorporated herein. PCT application PCT/USOO/02687, published as WO 00/47062, the contents of which are incorporated herein, also discloses a method of manufacturing a proanthocyanidin polymeric composition isolated from Croton spp. or Calophyllum spp., as well as methods of preparation. Another illustrative method for isolating proanthocyanidin polymer from C. lechleri (such as crofelemer) is found in U.S. Patent Nos. 7,341,744 and 7,323, 195, the contents of which are expressly incorporated herein. As described above, the SP 303 and SB 300 purified forms of oligomeric proanthocyanidin polymer from Croton lechleri in non-enteric forms, non-enteric formulations and compositions, are suitable for use in the treatment methods of the invention.

[0054] In an embodiment, the proanthocyanidin polymer composition may be generally isolated by the following process, such as provided in U.S. Patent No. 7,341,744, the contents of which are incorporated herein. Latex collected from Croton lechleri plants is mixed with purified water (preferably one part latex to two parts purified water). Any insoluble material in the latex solution is allowed to settle, e.g., by leaving the mixture at 4°C overnight (12 hours). The supernatant is pumped away from the residue and is extracted with a short chain alcohol, such as n-butanol. The extraction is preferably performed multiple times, such as three times. After each extraction, the alcohol phase is discarded and the aqueous phase is retained. The aqueous phase is concentrated, for example, using an ultrafiltration device with a 1 kD cut-off membrane. This membrane can be a low protein binding cellulose membrane, or, alternatively, a polypropylene, teflon or nylon membrane can be used. The membrane used should be compatible with acetone. The purpose of the ultrafiltration is to remove the water from the material.

[0055] The retentate from the ultrafiltration is then concentrated to dryness, for example using tray-dryers at approximately 37°C (± 2°C). The dried material is subsequently dissolved in water and is then chromatographed on a cation exchange column (e.g., a CM-Sepharose column) and a size exclusion column (e.g., an LH-20 column). In the preferred two column system, material is run over a CM-Sepharose and then an LH-20 column in a series. Specifically, the dissolved material is loaded onto the cation exchange column and is then washed with purified water. The proanthocyanidin polymer material is eluted from the cation exchange column with an aqueous acetone solution (preferably 30% acetone), thereby loading the proanthocyanidin polymer material onto the sizing column. The sizing column is disconnected from the cation exchange column and the material is then eluted off of the sizing column with an aqueous acetone solution (preferably 45% acetone). The fractions are collected and monitored with a UV detector, e.g., at a wavelength of 460 nm. Fractions containing the proanthocyanidin polymer material are combined and concentrated, for example, by ultrafiltration using, e.g., a 1 kD cut-off membrane (as described above for the ultrafiltration step prior to the chromatography steps). The retentate may then be concentrated to dryness using a suitable drying method, such as, but not limited to, a rotary evaporator, at a temperature of approximately 37°C (± 2°C). Other suitable drying methods include, but are not limited to, tray drying and spray drying. Example 10 of U.S. Patent No. 7,341,744 provides additional, non-limiting, methodology for preparing a composition comprising proanthocyanidin polymer. A detailed protocol for isolating an enriched proanthocyanidin polymer extract suitable for use in the methods of the invention is described in WO 00/47062 as noted herein above.

Components of pharmaceutically acceptable compositions and formulations for gastric ulcer treatment

[0056] Pharmaceutically acceptable preparations and formulations, particularly non- enterically coated or protected preparations and formulations, for stomach ulcer treatment as described herein may include, in addition to a Croton spp (e.g., C. lechleri)-derived proanthocyanidin polymer or botanical extract, any type of pharmaceutically acceptable excipients, additives, carriers, or vehicles. Such preparations and formulations do not include proton pump inhibitors or blockers, e.g., omeprazole, or H-2 receptor blockers. By way of nonlimiting example, diluents or fillers, such as dextrates, dicalcium phosphate, calcium sulfate, lactose, cellulose, kaolin, mannitol, sodium chloride, dry starch, sorbitol, sucrose, inositol, powdered sugar, bentonite, microcrystalline cellulose, or hydroxypropylmethylcellulose can be added to the non-enteric composition (e.g., a non-enteric C. lechleri proanthocyanidin polymer or extract composition) to increase the bulk of the composition. In addition, binders, such as, but not limited to, starch, gelatin, sucrose, glucose, dextrose, molasses, lactose, acacia gum, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapgol husks, carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, Veegum and starch arabogalactan, polyethylene glycol, ethylcellulose, and waxes, can be added to the formulation to increase its cohesive qualities. Further, lubricants, such as, but not limited to, talc, magnesium stearate, calcium stearate, stearic acid, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, carbowax, sodium lauryl sulfate and magnesium lauryl sulfate can be added to the formulation. Also, glidants, such as, but not limited to, colloidal silicon dioxide or talc can be added to improve the flow characteristics of a powdered formulation. Disintegrants, such as, but not limited to, starches, clays, celluloses, algins, gums, crosslinked polymers (e.g., croscarmelose, crospovidone, and sodium starch glycolate), Veegum, methylcellulose, agar, bentonite, cellulose and wood products, natural sponge, cation-exchange resins, alginic acid, guar gum, citrus pulp, carboxymethylcellulose, or sodium lauryl sulfate with starch can also be added to facilitate disintegration of the formulation at a treatment site.

[0057] In some embodiments, the non-enteric composition comprising C. lechleri proanthocyanidin polymer or extract may be formulated as an aqueous suspension in admixture with suitable excipients. Non-limiting examples of excipients that are suitable for the manufacture of aqueous suspension include suspending agents, for example, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethycellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents, which may be a naturally-occurring phosphatide, e.g., lecithin, or condensation products of an alkylene oxide with fatty acids, e.g., polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, e.g., heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol, for example, polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n- propyl, p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, e.g., sucrose, saccharin or aspartame. Dispersible powders and granules suitable for the preparation of an aqueous suspension by the addition of water can be used for components of the composition in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those stated above. Additional excipients, for example, sweetening, flavoring and coloring agents, may also be present.

[0058] In an embodiment, the stomach ulcer treatment composition including C. lechleri proanthocyanidin polymer or extract is in the form of a gel, paste, or gel paste formulation that is not enterically coated or protected, and which is contained or preloaded in a delivery device, such as a syringe, e.g., a needle-less syringe, or other type of applicator or delivery system, especially for oral delivery. In an embodiment, the gel is not contained in a delivery device, but is administered to the roof of the mouth of the animal, particularly one that may be too incapacitated or ill to eat or drink.

[0059] In a particular embodiment, a non-enterically coated or protected composition of

C. lechleri proanthocyanidin polymer or botanical extract is present in a gel or paste composition or formulation, preferably for oral administration. Other ingredients, such as cyto- or mucoprotective drugs or agents may be included in the composition or formulation. By way of example, the gel or paste may comprise, without limitation, an oily vehicle or excipient, such as a hydrophobic oily vehicle, a basifying agent, a flavoring agent and a coloring agent. Such a gel or paste is particularly suitable for oral administration. Illustrative and nonlimiting examples of hydrophobic oily vehicles include vegetable oil, triglyceride or polypropylene glycol, as well as a thickening agent, e.g., aluminum stearate. Flavoring agents can include, for example, fruit flavors, mint flavors, honey flavor, and other natural and organic flavorings known to those skilled in the art. Coloring agents can include, for example, iron oxide or titanium dioxide. Alternatively, the oily vehicle can be liquid paraffin or other suitable waxes, including a thickening agent. Oily suspensions may be formulated by suspending the non-enteric C. lechleri proanthocyanidin polymer or extract as active ingredient in a vegetable oil, e.g., arachis oil, olive oil, sesame oil or coconut oil, or in mineral oil, such as liquid paraffin. The oily suspensions may contain a thickening agent, e.g., beeswax, hard paraffin or cetyl alcohol. Oral preparations can include sweetening agents as mentioned above and flavoring agents to improve palatability. Pharmaceutically acceptable preservatives, for example, an anti-oxidant such as ascorbic acid, can also be added to such non-enteric compositions. No proton pump inhibitors or blockers, such as omeprazole, or H-2 receptor inhibitors, are employed in the compositions and methods of the invention.

[0060] The C. lechleri proanthocyanidin polymer or extract-containing, pharmaceutical compositions or formulations that are not enterically coated, as used in the methods of the invention, may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil such as olive oil or arachis oil, or a mineral oil such as liquid paraffin or mixtures of these oils. Examples of emulsifying agents include, without limitation, naturally-occurring phosphatides, e.g., soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, e.g., sorbitan monooleate, and condensation products of partial esters with ethylene oxide, e.g., polyoxyethylene sorbitan monooleate. Sweetening, coloring and flavoring agents can be included in the emulsions.

[0061] Syrups and elixirs containing a non-enteric composition or product comprising the

C. lechleri proanthocyanidin polymer or C. lechleri botanical extract may also can be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions may be in the form of a sterile, orally deliverable or administrable aqueous or oleagenous suspension. This suspension may be formulated according to methods known in the art using suitable dispersing or wetting agents and suspending agents, such as those mentioned above. The sterile pharmaceutical preparation may also be a sterile solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, a solution in 1,3 -butane diol. Illustrative, acceptable vehicles and solvents that may be used in the preparations include water, Ringer's solution and isotonic sodium chloride solution. Co-solvents, e.g., ethanol, propylene glycol or polyethylene glycols, may also be included. In addition, sterile, fixed oils, e.g., any bland, fixed oil such as synthetic mono- or diglycerides, are conventionally employed as solvents or suspending media and may be used. In addition, fatty acids, such as oleic acid and the like, may be used in injectable preparations.

Methods of Treatment and Applications of Use

[0062] The invention is directed to methods of treating and preventing stomach ulcers, especially stomach ulcers in young and adult animals, particularly equine animals like horses that can be naturally high-strung and can become stressed as a result of events in their habitats and lifestyles, as well as from endurance activities and performances expected of them. The methods also advantageously encompass the treatment of diarrhea that may be associated with gastric ulceratic conditions in afflicted animals. The method of the invention comprises administering to an animal in need of gastric ulcer treatment or prevention, a pharmaceutically acceptable, non- enterically coated or protected composition comprising a proanthocyanidin polymer or extract from a Croton species, in amounts effective to treat or prevent the gastric ulcers and/or the symptoms thereof. Advantageously, such compositions may also treat diarrheal conditions that may concurrently affect the animal. In preferred embodiments, the proanthocyanidin polymer or botanical extract is from a Croton species, namely, Croton lechleri and is a product that is not enterically coated or protected. Treating the gastric ulcers can involve reducing the severity and duration of the ulcers, including glandular and squamous ulcerations, in the animal. Treating the gastric ulcers can also involve decreasing, improving, or resolving the discomfort and pain associated with ulceratic lesions in the animal undergoing treatment. Treating the gastric ulceratic animals with compositions that are not enterically coated or protected and comprise a proanthocyanidin polymer or extract from Croton lechleri also results in treating a diarrheal condition that may result from or accompany the gastric ulceratic condition, as well as in improving the overall basic and gastrointestinal health of the animals, which may ultimately reduce their morbidity and mortality. As used herein, the adjectival term "non-enteric" is synonymous with "non-enteric coated or otherwise protected."

[0063] In an embodiment of the invention, the methods are directed to treating or preventing gastric ulcers in non-human young and adult animals suffering from gastric ulcers or at risk of developing gastric ulcers due to environmental conditions or lifestyle activities. The methods of the invention further relate to the treatment or prevention of gastric ulcers and/or related symptoms in adult, non-human animals, such as, without limitation, adult equine animals, exotic animals, and domestically or commercially used animals. Often, but not always, the animals are large in size and have complex gastrointestinal systems. For example, in horses, which are optimally suited for the treatment methods of the invention, the intestinal volume of the animal is large, with main sites of ulceratic lesions in the stomach.

[0064] In a related manner, ruminant animals, such as camels, which may also be treated by the methods of the invention as needed, possess multi-chambered stomachs, e.g. four stomach compartments, including a rumen or first compartment of the alimentary canal, which serves as the primary site for microbial fermentation of ingested feed. The described methods provide treatment of stomach ulcers in ruminant animals such as camels, sheep and goats with non- enterically coated compositions including a proanthocyanidin polymer or extract from C. lechleri, providing for an adequate amount and appropriate distribution of the non-enteric proanthocyanidin polymer or extract in the stomach of the animal, so as to treat gastric ulcers in an animal in need. [0065] As mentioned, the non-human young and adult animals that are susceptible to gastric ulcers for which the treatment methods are suitable may include different animal types, genera, or species. In general, young and adult farm animals, animals bred or kept for various purposes, such as sport (e.g., racing, riding, dressage), transport, domestic, companion, industrial uses (e.g. hauling, pulling, plowing), and the like, are particularly amenable to treatment according to the methods of the invention. For example, encompassed by the methods of the invention is the treatment of adult or young non-human animals, such as camels (calves), sheep (lambs), rams, horses (foals), pigs (piglets), goats (kids), bison/buffalo (calves), llamas, donkeys, mules, yaks, etc. Neonatal, young and adult exotic animals, such as zoo animals of various species, are also embraced by the treatments of the invention.

Dosage forms and administration

[0066] In an embodiment, the stomach ulcer treatment compositions that are not enterically coated or protected, comprising a C. lechleri proanthocyanidin polymer or extract is administered in a total dosage amount of 5 grams per day, or in an amount of 2.5 g twice per day, i.e., 2.5 g BID. In another embodiment, the stomach ulcer treatment compositions which are not enterically coated and which comprise a C. lechleri proanthocyanidin polymer or extract, is administered in a total dosage amount of 40 grams per day, or in an amount of 10 g four times per day, i.e., 10.0 g QID. In embodiments, the dosage is 0.5-2.0 mg/kg/day, or 10-15 mg/kg/day, or 70-100 mg/kg/day. In embodiments, the non-enteric composition comprises a C. lechleri proanthocyanidin polymer or extract which is SB 300, SP 303, or crofelemer. In a particular embodiment, the C. lechleri proanthocyanidin polymer or extract is SB 300, which is administered as a non-enterically coated or protected product. Those skilled in the art will appreciate that due to the higher purity of compositions such as SP-303 or crofelemer and SB- 300, more by weight of SB-300 than SP-303 will typically need to be used in compositions to achieve the same amount of the active ingredient of the non-enteric proanthocyanidin polymer or extract composition. SB-300 generally has about 67% by weight of the proanthocyanidin polymer composition while SP-303 has higher purity, for example 99-100%.

[0067] The routes of administration of compositions that are not enterically coated or protected and comprise a C. lechleri proanthocyanidin polymer- or extract to afflicted animals are not intended to be limiting. However, routes of administration amenable to optimally dispensing or administering an oral formulation, such as a paste, gel, or gel paste compositions, to the intended sites such as the stomach of the animal are preferred. By way of example, oral or buccal administration, including, without limitation, roof of mouth, dental, periodontal, or esophageal administration are encompassed by the invention. In embodiments, food source (animal feed), nutrition source, libation source, or food and/or drink supplement could be used. In an embodiment, the non-enteric coated or protected formulation or composition comprising a C. lechleri-denved proanthocyanidin polymer or extract SB-300 can be administered to an animal in need in the presence of feed. Such a mode of administration reflects a further advantage of the present gastric ulcer treatment methods over conventional methods involving a proton pump inhibitor such as omeprazole, which is typically administered to an animal without feed. Accordingly, the methods of the present invention do not impose constraints related to administration without feed and offer a superior treatment therapy for an animal in need.

[0068] In an embodiment, the non-enteric product could be provided in an aqueous formulation, administered to the animal as a drench or directly from a ready-to-use (RTU) bottle directed to the esophageal cavity so as to more effectively reach the animal's stomach or intestine for optimal activity. In a related embodiment, administration can also be by inclusion in the regular or special diet of the animal, such as in a functional food for the animals in need, or as a dietary supplement or food supplement, e.g., as described in WO 00/47062, for administration to an animal in need. In other embodiments, the non-enteric compositions of the invention can be formulated for rectal administration, such as a suppository, enema or other convenient form. The non-enteric compositions can also be provided as a controlled release system (See, e.g., Langer, 1990, Science 249: 1527-1533).

[0069] In other embodiments, for oral administration, the non-enterically coated or protected compositions or formulations comprising C. lechleri proanthocyanidin polymer or extract may be encapsulated and formulated with suitable carriers, and the like, in solid dosage forms. Nonlimiting examples of suitable carriers, excipients, diluents and vehicles include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, gelatin, syrup, methyl cellulose, methyl- and propylhydroxybenzoates, talc, magnesium, stearate, water, mineral oil, edible oils, and the like. The formulations can also include lubricating agents, wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents. As indicated, the non-enteric compositions may be formulated to provide rapid, sustained, extended, or delayed release of the active ingredients in the stomach or gastric region after administration to the animal by employing protocols and methods well known in the art. The non-enteric formulations and compositions may also include compounds or substances that reduce proteolytic degradation and promote absorption such as, for example, surface active agents.

[0070] As will be appreciated by those having skill in the art, the specific dose can be calculated according to the approximate body weight (e.g., mg/kg), body mass, or body surface area of the animal, or the volume of body space or mass to be occupied. The dose also depends on the particular route of administration selected by the practitioner. Further refinement of the calculations necessary to determine an appropriate dosage for treatment is routinely made by those of ordinary skill in the art, for example, using appropriate assays and analytical procedures, such as has been described for certain compounds (e.g., Howitz et al., 2003, Nature, 425: 191- 196). Exact dosages can be determined based on standard dose-response studies. Therapeutically effective doses for treatment of afflicted animals can be determined, by titrating the amount of the active product given to the animal to arrive at the desired therapeutic effect, while minimizing side effects.

[0071] For use in treating gastric ulcers and associated conditions in young animals, e.g., foals, in accordance with the methods of the invention, a therapeutically acceptable non-enteric form of the C. lechleri proanthocyanidin polymer or extract is administered, particularly orally administered, in an amount ranging from 0.1 to 100 mg/kg per day, once, twice, three times, four times, or more, daily. In other embodiments, the amount can range from about 0.1 to about 10 mg/kg/day, once, twice, three times, four times, or more daily; or from about 0.1 to about 25 mg/kg/day, once, twice, three times, four times, or more, daily; or from about 0.1 to about 30 mg/kg/day, once, twice, three times, four times, or more, daily; or from about 0.1 to about 40 mg/kg/day, once, twice, three times, four times, or more, daily. In other embodiments, the dose can be 0.1 mg/kg, 0.2 mg/kg, 0.3 mg/kg, 0.4 mg/kg, 0.5 mg/kg, 0.6 mg/kg, 0.7 mg/kg, 0.8 mg/kg, 0.9 mg/kg, 1 mg/kg, etc., as well as incremental dose amounts in between. In still other embodiments, the amount can range from about 1 to about 10 mg/kg/day once, twice, three times, four times, or more, daily; or from about 1 to about 5 mg/kg/day, from about 1 to about 8 mg/kg/day, from about 1 to about 10 mg/kg/day, or from about 2 to about 4 mg/kg/day once, twice, three times, four times, or more, daily. In other embodiments, the amount can range from about 0.5 to about 2.0 mg/kg/day or from about 10 to about 15 mg/kg/day, or from about 70 to 100 mg/kg/day, administered once, twice, three times, four times, or more, daily. In some embodiments, the foregoing amounts of the non-enterically coated compositions or formulations comprising a C. lechleri proanthocyanidin polymer or extract are administered, for example, twice daily, three times daily, four times daily, or more than four times daily, rather than once per day. Higher doses, e.g., 50 mg/kg or 100 mg/kg per day or twice or more daily, may be required, as necessary.

[0072] In other embodiments, for the treatment methods, a suitable dose for the C. lechleri proanthocyanidin polymer as a non-enterically coated product, or, in particular, the non- enteric C. lechleri extract as a non-enterically coated product, such as SP 303 or SB 300, may range from about 1 mg to about 1000 mg, or from about 10 mg to about 500 mg, or from about 50 mg to about 350 mg, or from about 30 mg to about 400 mg, or from about 100 mg to about 250 mg, or from about 50 mg to about 300 mg. It will be understood that the ranges include the lower and higher amounts specified, as well as amounts in between. The compositions can be administered once a day, or multiple times per day, as appropriate or required. Doses administered once or multiple times per day can be given for consecutive days, e.g., two days, three days, four days, five days, six, days, seven days, or more, in some embodiments. A dose administered multiple times per day may embrace two, three, four, five, six, ten, or more times per day. Other dosing schedules, such as every other day, or every third day, every fourth day, etc. are embraced by the invention. In addition, one having skill in the art will appreciate that doses and amounts administered to the animal can vary, given the wide range of weights of the animals undergoing treatment, as well as the animal species and type of digestive system, e.g., ruminant or non-ruminant.

[0073] In particular embodiments, the C. lechleri proanthocyanidin polymer or botanical extract is administered to an animal, such as an adult horse with gastric ulcerations, at a dose of 2.5 g BID, 2 doses per day, for a total of 5 g per day per animal, in a non-enterically coated or protected formulation. In other particular embodiments, the C. lechleri proanthocyanidin polymer or botanical extract is administered to an animal, such as an adult horse with gastric ulcerations, in a dose of 10 g QID, 4 doses per day, for a total of 40 g per day per animal, in a non-enterically coated or protected formulation. In an embodiment, the dose is from 0.5 to 2.0 mg/kg/day, or from 10 to 15 mg/kg/day, in one or multiple doses, such as two doses (BID). In an embodiment, the dose is from 70 to 100 mg/kg/day, in one or multiple doses, such as two doses (BID) or four doses (QID). In an embodiment, a composition that is not enterically protected comprises a C. lechleri proanthocyanidin polymer or botanical extract that is SB 300.

[0074] In some embodiments, total daily doses, including multiple daily doses, e.g., twice, three times, or four times per day, of the C. lechleri proanthocyanidin polymer or botanical extract as a non-enterically coated product, and amounts of the proanthocyanidin polymer or extract in the non-enteric compositions of the invention may be 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 50 mg., 100 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 500 mg, or 0.25 g, 0.5 g, 1 g, 2 g, 3 g, 4 g, 5 g, 10 g, 15 g, 20 g, 25 g, 30 g, 35 g, 40 g, 45 g, 50 g, 55 g, etc. (or amounts there between) per animal. By way of nonlimiting example, a total daily dose of 0.25 g, 0.5 g, 1 g, 2 g, 3 g, 4 g, 5 g, 10 g, 15 g, 20 g, 25 g, 30 g, 35 g, 40 g, 45 g, 50 g, 55 g, etc. (or amounts there between) of the C. lechleri proanthocyanidin polymer or extract product may be administered, e.g., orally, to an animal as a single dose, or may preferably be divided into multiple doses to achieve the aforementioned total dose, for example, once, twice, three times, four times, or more daily. Administration schedules may also be altered to achieve a therapeutically effective concentration of the non-enteric C. lechleri proanthocyanidin polymer or C. lechleri extract to treat gastric ulcers and ulcerations and their symptoms as described herein. By way of specific yet nonlimiting example, a suitable dosage amount of a composition or formulation that is not enterically coated and comprises a C. lechleri proanthocyanidin polymer or extract for use in the methods according to the invention is a total dose of 500 mg divided into 2 doses of 250 mg per day (BID); or a total dose of 5 grams divided into 2 doses per day; or a total dose of 40 grams divided into 4 doses per day. In an embodiment, the dose is 10-15 mg/kg/day or 70-100 mg/kg/day. In another specific embodiment, the dose of the C. lechleri proanthocyanidin polymer or C. lechleri botanical extract formulated as a non-enterically coated product is 2.5 g BID. In another specific embodiment, the dose of the C. lechleri proanthocyanidin polymer or C. lechleri botanical extract formulated as a non-enterically coated product is 10 g QID. In a specific embodiment, the C. lechleri proanthocyanidin polymer or C. lechleri botanical extract administered for treatment of gastric ulcers is SB 300 formulated as a non-enterically coated product.

[0075] It will be understood that the dose amount actually administered can be determined by the practitioner, in the light of the relevant circumstances, including the severity of the gastric ulcer condition, and/or symptoms thereof being treated, the age, weight, and response of the individual animal receiving treatment, as well as the chosen route of administration.

[0076] In other aspects, the methods of the present invention comprising treating gastric ulcers or EGUS in animals in need with a C. lechleri proanthocyanidin polymer or C. lechleri botanical extract formulated as a non-enterically coated or protected product, e.g., SB-300, may offer horse owners an additional advantage over conventionally-used treatments with proton pump inhibitors, such as omeprazole, in the competition horse world, where the requirement exists for equine athletes to compete free from the effect of any drugs. By way of example, international screening limits for horse racing state that omeprazole has a 72-hour detection time. Detection time is defined as the first observed time point at which urine and/or plasma samples collected from a horse are negative for the presence of a specified drug. Because the non- enterically coated SB 300 product acts locally in the gut and is minimally absorbed, it is likely that its detection time will be different and distinct from that of an inhibitor such as omeprazole. A non-enterically coated or protected formulation or composition comprising a C. lechleri proanthocyanidin polymer or C. lechleri botanical extract, such as SB-300, which may not be systemically absorbed, or not significantly systemically absorbed, in horses provides a gastric ulcer and EGUS treatment regimen that would be able to continue without mandatory withdrawal, or have a significantly reduced waiting time after treatment, by 12, 24, 36 or 48 hours, of an animal prior to competition. EXAMPLE

Example 1

Proof of Concept Field Study to Evaluate the Safety and Effectiveness of a Botanical Extract from Croton lechleri, SB-300, for the Treatment of Gastric Ulcers in Horses

Background

[0077] Endoscopic and post-mortem studies have demonstrated gastric ulcers in up to

93% of thoroughbred race horses in training. As mentioned hereinabove, stall confinement, stress, intermittent feeding, intense exercise and administration of non-steroidal antiinflammatories are factors that contribute or lead to gastric ulcers in horses. Gastric ulcers occur primarily in the nonglandular (squamous) portion of the stomach due to its lack of resistance to the erosive effects of gastric acids (hydrochloric, volatile fatty and bile acids). Horses with this condition may perform poorly, resulting in a significant economic problem within the horse industry.

[0078] Current therapy strategies are based on the reduction of gastric acidity and can be achieved through the use of a number of different drugs, e.g., antacids, H2-receptor antagonists (ranitidine and cimetidine), prostaglandin analogs (misoprostol) and proton pump inhibitors (omeprazole). However, long-term treatment with drugs that reduce the gastric acidity is expensive, and a prolonged increase in gastric fluid pH might have a negative effect on gastric physiology and digestion in affected animals. In addition, ulcer recurrence is reported to be common after discontinuation of treatment. The development of a natural alternative to maintain stomach health without altering stomach pH would be advantageous and desirable. SB 300 is a purified extract of the latex from Croton lechleri. The compound acts locally in the gastrointestinal tract with minimal systemic absorption, a key factor for the safety of the product. SB 300 has no effect on gut motility and no significant drug-drug interactions involving SB 300 have been identified to date.

Study objective

[0079] The primary objective of the proof of concept (POC) study was to investigate the safety and effectiveness of an orally administered paste formulation of SB-300 for the treatment of gastric ulcers in horses. In addition, the study tested two different doses of SB 300 to help characterize the dose for further clinical evaluation. The intention of the study protocol was exploratory, e.g., to gather data related to the effectiveness and outcome of treatment of gastric ulcers in horses with a non-enterically coated or protected SB 300 product as active.

Inclusion and exclusion criteria

[0080] The following horses satisfied the inclusion criteria for the study: (i) horses with the presence of gastric ulcers; (ii) horses with at least one squamous ulcer score of 2 or greater and/or at least one glandular ulcer that was either moderate or severe in severity; (iii) horses with concurrent chronic disease(s) were included in the study at the discretion of the Investigator if the condition had been stabilized and animals were medically well managed; (iv) horses that were in training. A horse was enrolled in the study and treatment was initiated if all of the inclusion criteria and none of the exclusion criteria (below) were met.

[0081] The following horses satisfied the exclusion criteria for the study: (i) horses that do not present with gastric ulcers; (ii) horses with squamous or glandular ulcers that did not meet the minimum requirements for study inclusion; (iii) horses with a positive fecal egg count at Screening; (iv) horses receiving prohibited medications or those that had failed to meet the washout periods for the medications; (v) horses with elevated fibrinogen and/or serum amyloid A levels that were considered clinically significant by the Investigator and indicative of a chronic pathologic condition; (vi) horses with other medical and/or behavioral conditions which in the opinion of the Investigator would preclude them from being enrolled into the study; and (vii) horses intended for breeding, or that were pregnant or lactating.

Study design and overview

[0082] The study was a prospective, masked, randomized, negative controlled, single site field study.

[0083] Horses suspected of having gastric ulcers were recruited and screened for the study. Screening procedures commenced following execution of the informed consent and included a medical and medication history, physical examination and body weight, clinical pathology testing (CBC, CHEM), fecal egg count, fecal pH, fecal blood test and gastroscopy to determine baseline gastric ulcer scores and gastric pH. For the study, male or female horses {Equus caballus, domestic horse) were used. The breed of horse included performance breeds prone to gastric ulcers. There were no limitations as to body weight, and the horses were adult horses greater than or equal to 2 years of age, with no upper limit. Also, the animals' reproductive status was intact or castrated. Horses meeting the enrollment criteria were randomized to 1 of 3 experimental groups and dosed with the Investigational Veterinary Product (IVP) either BID or QID or with the Control Product (CP) QID. The total daily dose was 5 g for the active BID group and 40 g for the QID group.

[0084] Dosing continued for 28 consecutive days. Horses remained on treatment for the full 28 days, regardless of whether there was evidence of complete resolution of their gastric ulcers. Clinical Trial Material (CTM) consisted of the IVP active product and was supplied as a flavored oral paste at a concentration of 10 g of SB 300 per 60 mL, in metered 60 mL dosing syringes for IVP. The Control Product (CP) was supplied as empty 60mL syringes and filled with water at the time of administration. The total daily dose was 5 g for the active BID group and 40 g for the QID group.

[0085] During the Treatment Period horses underwent repeated gastroscopy every 14 days

(Days 14 and 28) during which gastric ulcer scores and gastric pH were recorded. Fecal pH and the presence of fecal blood were recorded once weekly. Adverse events and concomitant medications were continuously monitored and recorded during this time period also. On Day 28, horses underwent a full evaluation which included, in addition to the gastroscopy and gastric pH, a physical examination and body weight, clinical pathology testing (CBC, CHEM), fecal pH and a fecal blood test. The last day of treatment was Day 28. To assess the treatment effect on gastric ulcers, one final gastroscopy on Day 35 was performed before the horse completed the study.

Number of animals/treatment group

[0086] Study-eligible horses were randomly assigned to 1 of 3 experimental treatment groups: IVP A, IVP B, or CP, as shown in the below table. The enrollment target was at least 10 evaluable cases per group for a total of 30 evaluable cases. Each individual horse was considered an experimental unit. [0087] Table 1

[0088] A single randomization schedule was prepared using SAS® Proc Plan. Horses were randomly assigned to an experimental group based on order of entry into the study. The randomization table was maintained in confidence by the site's Dispenser. The Dispenser stored all forms and other documents related to treatment in a secured location.

Clinical trial material (CTM) administration

[0089] If a horse was randomized to the Control group, in a blinded fashion, the dispenser prepared an empty 60 mL syringe with tap water and followed the instructions outlined below for administration. Each horse in the control group was given one syringe that was cleaned after each administration and reused throughout the duration of the study, unless the reuse of the syringe was contraindicated, and then a new syringe was utilized.

[0090] On Day 0, horses, by order of enrollment, were randomized to treatment. There were no special considerations regarding a fed or fasted state and doses were administered independent of the horse's feeding schedule. Treatments were administered using the following set of instructions:

• The correct number of syringes was selected based on treatment group assigned in the randomization schedule, and the horse's ID # and date were recorded on the syringe(s) using an indelible marker.

• The horse's mouth was cleared of any feed or hay that may have been present.

• The cover was removed from the tip of the syringe, and the syringe was inserted into the horse's mouth at the interdental space.

• The plunger was depressed until the entire dose had been given. The dose was to be deposited on the back of the tongue or deep into the cheek pouch.

• The horse was observed briefly after administration to ensure that all/part of the dose was not lost or rejected. If any of the dose was lost, re-dosing was recommended. The amount of any paste that was lost was estimated and recorded on the CRF.

• Syringes were not to be shared between horses.

• All used syringes were retained for drug reconciliation and accountability purposes.

• Prior to the gastroscopy procedure, feed and water were withheld for approximately 12 hours and 4 hours, respectively. To adhere to this requirement, treatment was also withheld prior to the scoping procedures, on Days 14 and 28, as needed, to enable full evaluation of the stomach during scoping. Treatment and food were withheld post procedure until the sedation wore off, approximately 2 hours post procedure. All attempts to dose accordingly were made post procedure. However, it was possible that 1 or 2 early day dosing for the QID group and 1 early day dosing for the BID group would be withheld. The withheld treatment timepoints were documented as "dose withheld due to gastroscopy."

Study practices

[0091] No changes were made to the horse's housing or management. Horses were permitted to maintain their training schedule for the duration of the study. However, they were not allowed to race or perform during this time period. Horses were maintained on their regular diet and feeding schedule for the duration of the study. The study required no special foods and no special feeding schedule. Water was available ad libitum except prior to the gastroscopic examination where feed and water were withheld for approximately 12 and 4 hours, respectively.

[0092] Current medications and those given to the horses within the last 14 days prior to enrollment in the study, as well as all concomitant medications administered during the study, were recorded. Medications that were prohibited for horses during the study and for 14 days prior to their first study treatment included all prescription, OTC, nutritional and homeopathic (herbal) anti-ulcer treatments: proton pump inhibitors (e.g. omeprazole), H2 antagonists (e.g. cimetidine, ranitidine), prostaglandin analogs (e.g. misoprostol), sucralfate, pre/probiotics, antacids; all ulcerogenic treatments: NSAIDS (e.g. flunixin, phenylbutazone); and any other medication that, in the opinion of the Investigator/Examining Veterinarian, could interfere with the study objectives.

Effectiveness variables

[0093] Effectiveness of treatment was determined based on several effectiveness variables. 1) Resolution of Gastric Ulcers: Resolution of gastric ulcers was defined as follows: Squamous ulcer: Grade 0; Glandular ulcer: Intact epithelium. Scarring and hyperemia may be present. 2) Improvement of Gastric Ulcers: Improvement of squamous ulcers was defined by a reduction in the Gastric Ulcer Council Scores by 1 grade or more (e.g., going from Grade 3 to Grade 2). Improvement of glandular ulcers was defined as a decrease in severity (e.g., going from moderate to mild) and improvement in description (e.g., going from raised and hemorrhagic to flat and hemorrhagic). 3) Time to Resolution of Gastric Ulcers: Time to resolution was determined by the elapsed time between the first dose and when the horse met the definition of gastric ulcer resolution. 4) Persistence of Treatment Effect: Persistence of treatment effect was determined by the reoccurrence of gastric ulcers at Day 35.

Gastroscopy

[0094] Horses underwent gastroscopic examination at Screening (baseline) and repeated prior to dosing on Days 14 (±1 day) and 28 (±1 day). A follow up examination was repeated at Day 35 (±1 day). In addition to the veterinarians performing the gastroscopy, an expert panel of equine veterinarians specializing in gastric ulcers and gastroscopy provided independent review of the endoscopic evaluations.

[0095] Prior to the procedure, feed and water were withheld for approximately 12 hours and 4 hours, respectively. During this time, hay was removed from the stall and the horse muzzled. For the procedure, horses were sedated (not anesthetized) according to the facility protocol and lightly restrained for the standing endoscopic examination (e.g., upper lip twitch). All medications used for the procedure were recorded on the CRE Once the endoscope was advanced into the stomach, the stomach was distended by insufflation of air through the endoscope until the gastric ruggae disappeared and the nonglandular, margo plicatus and glandular regions of the gastric surface could be easily observed. Gastric contents were then thoroughly rinsed from the stomach surface using tap water flushed through a pump.

[0096] The endoscopist ensured that both the glandular and nonglandular regions of the stomach were fully visible and that all ulcers present were able to be evaluated and scored. During the Day 0 gastroscopic examination, all ulcers that were present were evaluated and recorded. At the Day 14, 28 and 35 gastroscopic examination, all ulcers that were present were evaluated, including newly occurring ulcers that were not present at Day 0, and a single non- glandular score and a single glandular score were recorded. In addition to the completed CRFs, a video recording was made of each endoscopic procedure and filed within the study archives as raw data.

Evaluating the nonglandular region (squamous mucosa)

[0097] Ulcers were identified and mapped according to location within the squamous mucosa. Each squamous ulcer was scored according to the following scoring system:

Table 2 - Equine Gastric Ulcer Council 0-4 Scoring System*

* Sykes BW et al., European College of Equine Internal Medicine-Consensus

Statement, 2015 (unpublished draft version).

Evaluating the glandular region (glandular mucosa)

[0098] Ulcers were identified and each was evaluated based on the following criteria:

1. Anatomical location: (a) Cardia; (b) Fundus; (c) Antrum; and (d) Pylorus;

2. Description:

(a) Number: (i) Focal; (ii) Multi-focal; (iii) Diffuse;

(b) Severity: (i) Mild; (ii) Moderate; and (iii) Severe; and

(c) Description: (i) Flat and hemorrhagic; (ii) Flat and fibrinosuppurative; (iii) Raised and hemorrhagic: (iv) Raised and fibrinosuppurative; (v) Depressed ± blood clot; and (vi) Depressed and fibinosuppurative.

[0099] Gastric pH was measured during each gastroscopic examination at screening and on Day 14 (± 1 day) and Day 28 (± 1 day), and on Day 35. For measuring gastric pH, a sample of gastric juice was aspirated through the endoscope prior to insufflation of the stomach during the gastroscopy procedure. A handheld pH spear instrument was used to determine the pH. The instrument was cleaned and calibrated prior to each measurement. Measurement of gastric pH with pH electrodes in horses and foals and values thereof is also described in Murray, M.J. and Schusser, G.F., 1993, Equine Vet. J, 25(5):417-421 and in Murray, M.J. and Grodinsky, C, 1989, Equine Vet. J. Supplement, 7:73-76. In addition, fecal pH was measured at screening and weekly thereafter during the treatment period. For measuring fecal pH, a bowel movement within 15 minutes of evacuation was used. Five separate samplings were tested with a spear pH meter and the results averaged. For each reading the probe was inserted in the fecal matter, cleaned and calibrated prior to every reading.

Study results and summary

[0100] In this prospective, blinded, randomized, negative controlled study, Standardbred or Thoroughbred racehorses were randomized to one of three groups (10 horses per group) and treated for 28 days. The placebo or control group included horses that received water from water-filled syringes every 6 hours. The horses in the "TRT5" treatment group received 5 grams of SB-300 divided into 2 doses per day; and those in the "TRT40" treatment group received 40 grams of SB-300 divided into 4 doses per day. Strict enrollment criteria required that horses in the study had both squamous (non-glandular) and glandular gastric ulcerations. All horses were examined by gastroscopy (stomach endoscope) by blinded experienced equine investigators on Day 0 (prior to treatment; baseline), and on Day 14 (mid-study), Day 28 (last day of treatment) and Day 35 (7 days after last treatment). Treatment-related adverse events were not observed.

[0101] With respect to glandular ulcerations, a statistically significantly greater number of horses in both the TRT40 (89%) and the TRT5 (78%) treatment groups had an improvement or a resolution of glandular ulcerations, compared with the placebo (25%) group as soon as Day 14 during treatment. By Day 35 of the study, all of the SB 300-treated horses had experienced improvement or resolution, whereas 25% of horses in the placebo group still had not improved or resolved during the study. By contrast, only between 14% and 34% of horses diagnosed with EGUS and treated with the proton pump inhibitor omeprazole showed resolution or improvement of glandular ulcers when used at the manufacturer's recommended treatment duration of 28 days. (Sykes, B.W. et al., 2015, Equine Vet. J, 47(3):285-290; Sykes, B.W. et al., 2014, Equine Vet. J, 46(5):416-421). [0102] With respect to squamous ulcerations, a non-statistically significant dose- dependent effect was observed with 40 and 33% of horses resolving by Day 14 in the TRT40 and TRT5 treatment groups, respectively, compared with 11% of placebo horses. By Day 35, numerically more horses in the TRT40 (60%) and TRT5 (55%) treatment groups had resolved compared with 33% of placebo horses. Results are presented in Table 3 :

Table 3 - Gastric Ulcer Effectiveness Variables Per Protocol Population

Pairwise Comparisons

2.5 g BID 10 g QID Fisher's Exact BID vs QID vs

Ulcer Type Parameter Study Day CP (N=9) (N=9) (N=10) (2-tail) p-value Control Control

Non-Glandular Resolution (Grade=0) Day 14 1 (11 .1 %) 3 (33.3%) 4 (40.0%) 0.4454 0.5765 0.3034

Day 28 3 (33.3%) 3 (33.3%) 4 (40.0%) 1 .0000 1 .0000 1.0000

Day 35 2 (22.2%) 4 (44.4%) 2 (20.0%) 0.5768 0.6199 1.0000

Improvement Day 14 9 (100.0%) 9 (100.0%) 10 (100.0%)

Day 28 9 (100.0%) 7 (77.8%) 10 (100.0%) 0.1905 0.4706

Day 35 9 (100.0%) 6 (66.7%) 10 (100.0%) 0.0513 0.2059

Persistence of Treatment Day 35 2 (22.2%) 3 (33.3%) 2 (20.0%) 0.8687 1 .0000 1.0000 Effect

Time to Resolution Day 14 1 (11 .1 %) 3 (33.3%) 4 (40.0%) 0.6555 0.5335 0.4090

Day 28 2 (22.2%) 1 (11 .1 %) 2 (20.0%)

Day 35 0 (0.0%) 1 (11 .1 %) 0 (0.0%)

Did not resolve 6 (66.7%) 4 (44.4%) 4 (40.0%)

2.5 g BID Fisher's Exact BID vs QID vs

Ulcer Type Parameter Study Day CP (N=8) (N=9) 10 g QID (N=9) (2-tail) p-value Control Control

Glandular Resolution (Grade=0 or 1) Day 14 1 (12.5%) 8 (88.9%) 8 (88.9%) 0.0005 0.0034 0.0034

Day 28 4 (50.0%) 5 (55.6%) 8 (88.9%) 0.2096 1 .0000 0.1312

Day 35 3 (37.5%) 6 (66.7%) 8 (88.9%) 0.0949 0.3469 0.0498

Improvement Day 14 6 (75.0%) 9 (100.0%) 9 (100.0%) 0.0862 0.2059 0.2059

Day 28 8 (100.0%) 9 (100.0%) 9 (100.0%)

Day 35 8 (100.0%) 8 (88.9%) 9 (100.0%) 1 .0000 1 .0000

Persistence of Treatment Day 35 2 (25.0%) 6 (66.7%) 8 (88.9%) 0.0321 0.1534 0.0152 Effect

Time to Resolution Day 14 1 (12.5%) 8 (88.9%) 8 (88.9%) 0.0020 0.0063 0.0063

Day 28 4 (50.0%) 1 (11 .1 %) 1 (11 .1 %)

Day 35 1 (12.5%) 0 (0.0%) 0 (0.0%)

Pairwise Comparisons

2.5 g BID 10 g QID Fisher's Exact BID vs QID vs

Ulcer Type Parameter Study Day CP (N=9) (N=9) (N=10) (2-tail) p-value Control Control

Did not resolve 2 (25.0%) 0 (0.0%) 0 (0.0%)

Resolution (Grade=0) Day 14 1 (11 .1 %) 3 (33.3%) 6 (60.0%) 0.1216 0.5765 0.0573

Day 28 1 (11 .1 %) 4 (44.4%) 5 (50.0%) 0.2222 0.2941 0.1409

Day 35 2 (22.2%) 5 (55.6%) 4 (40.0%) 0.3967 0.3348 0.6285

[0103] Analysis of the study results indicated that SB-300 did not alter gastric pH during the 28-day trial, or for 7 days after therapy. Gastric pH during therapy was observed to be similar to baseline gastric pH (pH at screening) at all measured study time points. While other ulcer treatments, e.g., proton pump inhibitors like omeprazole, act by blocking gastric acid section for the treatment and prevention of EGUS, the data from the present study indicate that SB-300 may have advantages over known gastric acid inhibitors by not altering or affecting the animal's gastric pH. Treatments for EGUS that do not alter gastric pH are important because maintaining low gastric pH is essential for the animal's digestion, for gut immunity and first line defense against pathogens, for the absorption of vitamins and minerals, as well as for other potential downstream effects.

[0104] All patents, patent applications and publications referred to or cited herein are hereby incorporated by reference in their entireties for all purposes.

[0105] It is understood that the embodiments and examples described herein are for illustrative purposes and that various modifications or changes in light thereof will be suggested to persons skilled in the pertinent art and are to be included within the spirit and purview of this application and scope of the appended claims. It is to be understood that suitable methods and materials are described herein for the practice of the embodiments; however, methods and materials that are similar or equivalent to those described herein can be used in the practice or testing of the invention and described embodiments.