WO2013132092A1

WO2013132092A1 - Use of monounsaturated fatty acid compounds for controlling the body weight of a dog or a cat

Info

Publication number: WO2013132092A1
Application number: PCT/EP2013/054807
Authority: WO
Inventors: Mikael S. THOMSEN; Søren STENDERUP; Jenny HÄGGLÖF
Original assignee: Pensieve International Plc
Priority date: 2012-03-09
Filing date: 2013-03-11
Publication date: 2013-09-12

Abstract

Use of a compound of the general formula (I), COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein R can be any group with a molecular weight from 14 to 200 Da, m and n are independently selected from an integer of 0 to 15, and the double bond is in cis or trans configuration or a mixture thereof, or a veterinary acceptable salt, ester or glyceride thereof, for controlling body weight in a pet selected from the group consisting of a dog and a cat.

Description

USE OF MONOUNSATURATED FATTY ACID COMPOUNDS FOR

CONTROLLING THE BODY WEIGHT OF A DOG OR A CAT

Field of invention

The present invention relates to the use of 2-hydroxyoleic acid and related fatty acid compounds, or veterinary acceptable salts, esters or glycerides thereof, for use in the treatment of a pet, such as a dog or a cat. In particular, the present invention relates to the use of 2-hydroxyoleic acid or veterinary acceptable salts, esters or glycerides thereof, for use in body weight management of an obese subject, such as an obese dog or an obese cat.

Background of invention

Obesity in pets is common in many countries. An estimated 54% of dogs and cats in the United States are overweight or obese. In absolute terms this corresponds to an estimated 93 million US dogs and cats.

The reasons for obesity of cats and dogs in wealthier countries are not simply a matter of overfeeding. Lifestyle influences also play roles. In modern times, pets are often, for their own safety, not allowed to free roam as they did in the past. Modern day pet foods are a higher quality food, and dogs, as natural scavengers, are prone to gorging themselves whenever the opportunity arises, with the limit being stomach capacity rather than hunger level. In evolutionary past, this was the only way to survive as they wouldn't know where their next meal would be coming from. More recently, pets, such as dogs and cats, and working dogs were fed on scraps of whatever was left over in the household modern pet food is nutritious. However, owners are prone to overfeeding dogs and cats, as they see the bowl of food and think that it looks like not enough. The taste of the food encourages dogs and cats to want to eat more.

Modern lifestyles often mean that owners are not able to find the time to walk their dog for an hour or more a day, thus reduced exercise, coupled with richer foods and owners who believe that they are doing the right thing by overfeeding, are the main cause of obesity illness in pets today. Further many cats are kept indoor as traffic in densely populated areas can expose a freely roaming cat to dangers.

Obesity is usually accompanied by a wide array of divergent health disorders including respiratory problems such as astma, certain types of cancer (e.g. kidney, colorectal, prostate, ovarian, uterine/endometrial, esophaegal, pancreatic and port-menopausal breast), type 2 diabetes, cardiovascular diseases (e.g. hypertension, coronary artery disease, congestive heart failure, pulmonary embolism, stroke, dyslipidaemia), gall bladder disease, chronic back pain, osteoarthritis and sleep apnea which finally contribute to the described reduction in life expectancy. The severity of the overweight or obesity is correlated with the frequency and severity of the interrelated diseases.

More specifically, obesity typically promotes the appearance of a cluster of other interrelated diseases, in particular cardiovascular diseases, e.g., hypertension, atherogenic dyslipidemia, insulin resistance and type 2 diabetes. Insulin resistance is the condition in which normal amounts of insulin are inadequate to produce a normal insulin response from fat, muscle and liver cells resulting in elevated blood glucose levels. Type 2 diabetes, is characterized by persistent hyperglycemia from any of several causes and is the most prominent disease related to failure of blood sugar regulation. High blood glucose levels subsequently promote a number of secondary, irreversible health dysfunctions, for example vascular macro- and microangiopathy, and for that reason, it is of high importance to avoid incidences of hyperglycemia.

Obesity has been found to contribute to approximately 55% of cases of type 2 diabetes.

Fatty acids are aliphatic mono-carboxylic acids derived from, or contained in esterified form in, an animal or vegetable fat, oil, or wax. Natural fatty acids commonly have a chain of 4 to 28 carbons, which may be saturated or unsaturated. Saturated fatty acids are long-chain carboxylic acids that usually have between 12 and 24 carbon atoms and no double bonds. Unsaturated fatty acids are of similar form, except that one

(monounsaturated fatty acid, MUFA) or more (polyunsaturated fatty acid, PUFA) alkenyl functional groups exist along the chain, with each alkene substituting a single- bonded " -CH2-CH2-" part of the chain with a double-bonded "-CH=CH-" portion (that is, a carbon double-bonded to another carbon). Polyunsaturated fat can be found mostly in grain products, fish and sea-food (herring, salmon, mackerel, and halibut), soybeans, and fish oil. Monounsaturated fats are found in natural foods such as nuts and avocados, and are the main component of olive oil. Common monounsaturated fatty acids are palmitoleic acid, cis-vaccenic acid and oleic acid. Fatty acids are

commercially used for soaps, synthetic organic detergents, lubricating greases, surface coatings, emulsifiers, cosmetics and in the food industry. International patent application no. WO 2005/041691 is directed to the use of hydroxyoleic acid and related compounds as functional food additives and/or ingredients, focusing on improving parameters related to cardiovascular diseases, such as hypertension and obesity. The object of the invention disclosed in this document is to claim the effect of prevention of cardiovascular diseases and obesity of hydroxyoleic acid and similar compounds, as nutritional dietary supplements. The document further teaches that fatty acids of the 2-hydroxyoleic acid type and their functional analogs produce effects of satiety, including reductions in food intake. This control is also mediated by receptors of cytokines, leptins, adrenoceptors and other receptors coupled to G-proteins, whose activity is said to be modulated by these fatty acids. In the experimental part, it is shown that non-obese rats and non-obese adult mice lost weight when receiving nutritional supplements of hydroxyoleic acid despite having free access to food and water. The document is silent with information as to whether a weight reducing effect might also be seen when 2-hydroxyoleic acid is given to higher ranking obese animals, such as cats and dogs.

European patent application no. EP 1435235 describes the use of hydroxyoleic acid and similar compounds in the production of medicaments as antitumor agents, as agents with hypotensive activity and as agents for inducing reductions in body weight. In the experimental part, it is shown that non-obese rats and non-obese adult mice lost weight when receiving nutritional supplements of hydroxyoleic acid despite having free access to food and water. The document teaches that 2-hydroxyoleic acid and its analogs produce effects of satiety, including reductions in food intake. This control is also mediated by receptors of cytokines, leptins, adrenoceptors and other receptors coupled to G-proteins, whose activity is modulated by these fatty acids. The document is silent with information as to whether a weight reducing effect might also be seen when 2-hydroxyoleic acid is given to higher ranking obese animals, such as cats and dogs.

International patent application no. WO 2010/066931 discloses a number of fatty acids, including 2-hydroxyoleic acid, to be used as medicines for the prevention and treatment of cardiovascular diseases and obesity and for the treatment of lung, brain or prostate cancer. In the experimental part of the document it is shown that rats pretreated with fatty acids showed significant less weight gain compared to rats receiving identical cafeteria diet. Hence, the document fails to show that a weight decrease is obtainable by giving 2-hydroxyoleic acid to an animal. This document is also silent with regard to information as to whether similar results could be obtained if experiments were performed on higher ranking animals such as cats and dogs.

International application no. WO 02/00042 is directed to compositions and methods for body weight management, particular weight gain and loss, which is effectuated by producing a sensation of satiety in subjects. Oleic acid is mentioned as one agent among a large number of agents, which may induce satiety. The document is silent with regard to any similar activity of 2-hydroxyoleic acid. The document discloses how to prepare ready-to-drink beverages, a confectionary bar, a molded tablet, a soft gelatin capsule, a hard gelatin capsule, a pharmaceutical emulsion and microcapsules. The document does not disclose any experiments showing the claimed effect of weight gain and loss.

It is well-known that it is difficult, if not impossible, to predict a drug response for an animal without performing a specific test on said animal. For example in Gauthier et. al. (The Journal of Pharmacology and Experimental Therapeutics, 1999, 687-693), the effect of various beta-adrenoceptor agonists on the contractility of ventricular strips sampled from various mammalian species was investigated. They found three groups of responders, which could be defined as hyper-responders (humans and dogs), hypo- responders (rat and guinea pig) and non-responders (ferret) to agonist stimulation in the myocardium. It is concluded that cautions should be taken in using animal models for the development of therapeutic compounds active on humans. Similar results is found for example in Baroni et al., (Regulatory Toxicology and Pharmacology, 2008,

332-341 ), where a study reveals a > 1000-fold higher sensitivity of dog compared to rat and human in induction of hypercalcemia and consequent systemic toxicity when administered 1 ,25-dihydroxyvitamin D₃. Furthermore, it is known from the marketing authorization of the weight loss drug Slentrol, which is approved for dogs only, that this drug must not be administered to cats due to the risk of development of hepatic lipidosis.

Another example supporting the non-predictability of a drug response is related to the drug Losartan. Following the synthesis of Losartan, pharmacological studies in rats revealed that an active, highly potent metabolite was created. It had a beneficial effect. It worked just like the drug, but it was much more powerful. Further, its effects lasted for a very long time. As a result, the metabolite amplified the antagonistic effect of Losartan. It was fortunate. It need not have been so. Troublingly though, animal tests showed that the metabolite's creation varied by species. It was created in rats but not in dogs. Naturally, a major concern going into clinical trials was whether the metabolite would be created in humans and what affects it would have.

The mode of action of fatty acids, such as 2-hydroxyoleic acid, is not known. As a consequence, it is not possible to predict whether positive results showing a weight reducing effect in a rodent, would also show a similar positive effect in a higher ranking animal such as for example a cat or a dog. Until today, fatty acids including 2- hydroxyoleic acid or a derivative thereof have not been considered for use in weight control of an obese or non-obese animal such as a cat or a dog as a pet food additive. Inventors of the present invention have realised that 2-hydroxyoleic acid may act as a weight reducing agent when fed to an obese cat or an obese dog as a food additive.

In 2007 the drug Slentrol was approved as a weight loss drug for dogs. The active substance in Slentrol is Dirlotapide. This drug, however, is not approved for cats. The following contraindications can be found in the product resume of marketing authorization of Slentrol: Do not use in dogs in growth phase, do not use in pregnant or lactating dogs, do not use in animals with impaired liver function, do not use in case of hypersensitivity to the active substance or excipient, do not use in animals in which overweight or obesity is caused by a concomitant systemic disease such as hypothyroidism or hyperadrenocorticism, and do not use in cats due to the risk of development of hepatic lipidosis.

Yarvitan was another drug for use in the management of overweight and obesity in adult dogs. The active substance of Yarvitan is Mitratapide. This drug was also only approved for dogs. In the product resume of the now withdrawn marketing

authorization of Yarvitan the following contraindications can be found: do not use in dogs with impaired liver function, do not use in case of hypersensitivity to the active substance or to any of the excipients, do not use in dogs during pregnancy and lactation, do not use in dogs less than 18 months of age, do not use in dogs in which overweight or obesity is caused by a concomitant systemic disease such as hypothyroidism or hyperadrenocorticism. This medicinal product is no longer authorized.

Hence, there exist a need for a new drug or food additive, which is safe to use for maintaining or reducing body weight in pets in general, such as in both a cat and in a dog in particular, and which is less toxic than the approved drug Slentrol and formerly approved Yarvitan. The present invention provides the solution to this problem.

Surprisingly, the inventors of the present invention has found that when obese dogs are administered 2-hydroxyoleic acid at a daily dosage of 200 mg/kg body weight a markedly weight loss is observed as compared with dogs treated with vehicle.

Moreover, the inventors of the present invention has found that cats in fact do tolerate 2-hydroxyoleic acid in that no systemic toxicity was observed in a study, where cats were administered up to 300 mg 2-hydroxyoleic acid/kg body weight with

corresponding (high) exposure.

A first strategy of preventing weight gain or providing weight loss in a pet is to mix the 2-hydroxyoleic acid and/or one or more related fatty acid compounds thereof with the pet food, and thereby feeding a recommended supplement to the pet in each meal.

In an alternative strategy, the 2-hydroxyoleic acid and/or one or more related fatty acid compounds thereof may be incorporated into the pet food by the manufacturer as part of the manufacturing process. In yet another strategy, the 2-hydroxyoleic acid and/or one or more related fatty acid compounds thereof may be mixed with the pet's drinking water.

In yet another strategy the 2-hydroxyoleic acid may be formulated in tablets, suspensions or capsules for oral intake in conjunction with or separate to food intake.

Summary of invention

The problem to be solved by the present invention is to provide a compound for use in the treatment of a pet selected from the group consisting of a dog and a cat, such as in the treatment of an obese subject selected from the group consisting of an obese dog and an obese cat. Accordingly, a first aspect of the invention relates to a compound of the general formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

R represents a group selected from H, OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN, N0₂, or some other group with a molecular weight from 14 to 200 Da

m and n are independently selected from an integer of 0 to 15; and

- the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

for use in the treatment of a pet selected from the group consisting of a dog and a cat.

The compounds designated by the general formula (I) is also referred to as fatty acid compounds throughout this application.

In a particular embodiment of the first aspect the invention relates to 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof, for use in body weight management of an obese subject selected from the group consisting of an obese dog and an obese cat.

A second aspect of the invention relates to a pet food supplement for feeding a pet, said pet food supplement comprising a compound of the general formula (I): COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

- m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

where the compound of formula (I) is used as a pet food additive for controlling body weight of a pet selected from the group consisting of a dog and a cat. In a particular embodiment of the second aspect, the present invention relates to a pet food supplement for feeding an obese subject, said pet food supplement comprising 2- hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof, where said 2-hydroxyoleic acid or veterinary acceptable salt, ester or glyceride thereof is used as a pet food additive for body weight management of an obese subject selected from the group consisting of an obese dog and an obese cat.

A third aspect of the invention relates to pet food for feeding a pet, said pet food comprising a compound of the general formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

R represents a group selected from H, OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN, N0₂, or some other group with a molecular weight from 14 to 200 Da,

m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

where the compound of formula (I) is used as a pet food additive for controlling body weight of a pet selected from the group consisting of a dog and a cat.

In a particular embodiment of the third aspect, the invention relates to a pet food for feeding an obese subject, said pet food comprising 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof, where said 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof is used as a pet food additive for body weight management of an obese subject selected from the group consisting of an obese dog and an obese cat.

A forth aspect of the invention relates to a method of controlling body weight of a pet, comprising administering to a pet an effective amount of a compound of formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein R represents a group selected from H, OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C₂-e alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN, N0₂, or some other group with a molecular weight from 14 to 200 Da

m and n are independently selected from an integer of 0 to 15; and

- the double bond is in cis or trans configuration,

or a veterinary acceptable, ester or glyceride salt thereof,

and where the pet is selected from the group consisting of a dog and a cat.

In a particular embodiment of the forth aspect, the present invention relates to a method of body weight management of an obese subject, comprising administering to said obese subject an effective amount of 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof, where the obese subject is selected from the group consisting of an obese dog and an obese cat. Definitions

Prior to a discussion of the detailed embodiments of the invention is provided a definition of specific terms related to the main aspects and embodiments of the invention. The term "general formula (I)" means a compound with a structure such as COOH-

CHR-(CH2)m-CH=CH-(CH2)n-CH₃ in which R can be any group with a molecular weight from 14 to 200 Da, but preferably R is selected from H, OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN or N0₂, m and n are

independently selected from an integer of 0 - 15, and the double bond is in cis or trans configuration, preferably in the cis configuration, or a veterinary acceptable salt, ester or glyceride thereof. In some embodiments R is not H.

The term "alkyl", as used herein, unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight or branched moieties. Examples of alkyl moieties include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and neopentyl. Alkyl is preferably Ci_-6 alkyl, i.e. groups containing from 1 to 6 carbon atoms, and for some embodiments of the present invention, more preferably Ci_-3 alkyl. The term "alkenyl", as used herein, unless otherwise indicated, includes alkyl moieties having at least one carbon-carbon double bond wherein alkyl is as defined above. Examples of alkenyl include, but are not limited to, ethenyl, propenyl, 1 -butenyl, and 2- butenyl. Alkenyl is preferably C_2-6 alkyl, i.e. groups containing from 2 to 6 carbon atoms, and for some embodiments of the present invention, more preferably C_2-3 alkenyl.

The term "alkynyl", as used herein, unless otherwise indicated, includes alkyl moieties having at least one carbon-carbon triple bond wherein alkyl is as defined above.

Examples of alkynyl groups include, but are not limited to, ethynyl, 2-propynyl, 1 - butynyl, and 2-butynyl.

The term "alkoxy", as used herein, means an -O-alkyl group wherein "alkyl" is as defined above. Alkoxy furthermore refers to polyethers such as -0-(CH₂)i-6-0-CH_3. Examples include, but are not limited to methoxy, ethoxy, propoxy, isopropoxy, n- butoxy, sec-butoxy, tert-butoxy, pentoxy, 2-pentyloxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy.

Any of the alkyl, alkenyl, alkynyl and alkoxy groups as defined herein may optionally be substituted with one or more of substituents selected from -F, -CI, -CF₃, -CCI₃, -CH₃, - OH, -CN, -N0₂, -NH₂ and methoxy.

The term "amino group" as used herein includes NH₂, primary amino groups, such as methylamino and ethylamino, secondary amino groups, such as dimethylamino and diethylamino, and tertiary amino groups, such as trimethylamino.

The term "2-hydroxyoleic acid" (2-OHOA) means octadecenoic acid C18:1 cisA9 or cis- 2-hydroxy-9-octadecenoic acid and is schematically represented below.

The term "2-hydroxyoleic acid" means 2-hydroxy-cis-9-octadecenoic acid. The term "2-methyl-oleic acid" means 2-methyl-cis-9-octadecenoic acid.

The term "2-amino-oleic acid" means 2-amino-cis-9-octadecenoic acid. The term "oleic acid" means cis-9-octadecenoic acid.

id:

The term "derivative" or "derivatives" means those fatty acids that have the double bond shifted one or two positions from the central zone and/or that have the double bond shifted from one to five positions from the central zone and/or have from one to four carbon atoms (CH₂ groups) more or less on each side of the double bond and/or that have a residue (R) in position 2 different from OH, with a small atomic mass (Mw less than or equal to 200 Da), and the stereoisomer corresponding to the projection of the R group in general formula (I) is in R- or S-configuration or racemic mixtures thereof.

The term an "effective amount" of 2-hydroxyoleic acid or a compound of general formula (I), as used herein, means an amount sufficient to control the body weight of a pet, either by maintaining the body weight of a non-obese pet or by reducing the body weight of an obese pet. Effective amounts for each purpose will depend on the weight of the pet as well as the race of the pet. The term "control the body weight" or "controlling the body weight", as used herein, means the management and care of a pet, which may be an obese or a non-obese pet, in order to secure the standard weight of a pet by feeding to the pet a pet food additive, which herein is designated by 2-hydroxyoleic acid or a compound of formula (I). The term is intended to include the full spectrum of controlling the body weight. More particular the term refers to maintaining the body weight of a non-obese pet and to reducing body weight in an obese pet.

The term "obese pet", as used herein, means a pet having an abnormal or increased body weight that may impair the pet's health. The term "non-obese pet", as used herein, means a pet, which is not obese, i.e. a pet that does not have an abnormal or excessive fat accumulation that may impair the pet's health. Body Condition Scoring (BCS) is an objective system of evaluating an animal's level of body condition (amount of stored fat) and may also be used in the present application to distinguish an obese pet from a non-obese pet.

The term "a veterinary acceptable salt" is intended to indicate salts, which are not harmful to the pet. Such salts are generally prepared by reacting the free base with a suitable organic or inorganic acid or by reacting the free acid with a suitable organic or inorganic base. When a compound according to the present invention comprises a free base (e.g. -NH₂) such salts are prepared in a conventional manner by treating a solution or suspension of the compound with a chemical equivalent of a veterinary acceptable acid. When a compound according to the present invention comprises a free acid (e.g. -COOH) such salts are prepared in a conventional manner by treating a solution or suspension of the compound with a chemical equivalent of a veterinary acceptable base. The term "a veterinary acceptable ester" is intended to indicate the product where 2- hydroxyoleic acid or an acid of general formula (I) has reacted with an alcohol and formed the resulting ester compound. Preferred alcohols that can be used for producing such an ester compound include, but are not limited to, methanol, ethanol, n- propanol and isopropanol.

The term "a veterinary acceptable glyceride" is intended to indicate the product where at least one acid of general formula (I), such as 2-hydroxyoleic acid, has reacted with glycerol and formed the resulting ester compound. The term includes mono-, di- and triglycerides indicating that one, two or three fatty acids have reacted with the glycerol. It should be understood that the scope of the present invention includes compounds, for which at least one acid of general formula (I) has reacted with the glycerol. Hence, a triglyceride, which for example has been produced by reacting glycerol with one acid of general formula (I) and two acids, which are different from the general formula (I), is also encompassed in the present invention by the term "a veterinary acceptable glyceride".

The term "pharmaceutical composition" is to be understood as a wide variety of pharmaceutical acceptable formulations and may be combined with one or more physiologically acceptable carriers. The pharmaceutical carrier or diluent employed may be a conventional solid or liquid carrier or diluent. Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, sterols, fatty acids, fatty acid amines, polyoxyethylene, isotonic buffer solutions and water.

The term "treatment" and "treating" as used herein means the management and care of a pet for the purpose of combating a condition, such as a disease or a disorder. The term is intended to include the full spectrum of treatments for a given disorder or condition from which the pet is suffering by administration of the 2-hydroxyoleic acid or a compound of general formula (I), or a veterinary acceptable salt, ester or glyceride thereof, thereof. More particularly, the term refers to alleviate or relief the symptoms and complications associated with the control of body weight by feeding to a subject the 2-hydroxyoleic acid or a compound of general formula (I), or a veterinary acceptable salt, ester or glyceride thereof, whereby the bodyweight is maintained or reduced.

Within the present context, the term "pet" embrace all kinds of animals that are fed by human beings. Pets may be kept in homes for cosy atmosphere, or for hunting or for rounding up cattle and sheep and the like or used as assistance by various forces such as police or customs or help to visually impaired persons. In particular the term embraces all kinds of dogs and cats. The term "pet food", as used herein, means a food that is produced for the purpose of feeding it to a pet as a meal. The term includes all kinds of pet food, which is suitable for comprising the additive of the present invention, but in particular the term "pet food" means both dry pet food and wet pet food. A pet food according to the present invention comprises, as a pet food additive, 2-hydroxyoleic acid or a compound having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof. It lies within the scope of the present invention that the pet food of the invention may also comprise a mixture of two or more different compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof. In particular, the mixture may comprise 2-hydroxyoleic acid and one or more compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof.

The term "pet food additive", as used herein, means an additive that may be added to a pet food or a pet food supplement in order to control the weight of the pet. In the present invention the pet food additive is described by 2-hydroxyoleic acid or by the compound having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof.

The term "pet food supplement", as used herein, means a separate food product that is produced for the purpose of feeding it to a pet together with the pet food or by mixing it with the pet's drinking water. The pet food supplement comprises, as a pet food additive, 2-hydroxyoleic acid or a compound having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof. It lies within the scope of the present invention that the pet food supplement of the invention may also comprise a mixture of two or more different compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof. In particular, the pet food supplement may be a mixture comprising 2-hydroxyoleic acid and one or more compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof.

In the context of the present invention, the term "daily dosage" is meant to describe the daily dosage required for a pet. It is well known that the daily dosage will depend on the type of pet, the weight of the pet and the pet's digestion and level of activity. In general, the daily dosage of the pet food supplement lies in the range of 1 - 400 mg/kg body weight.

Detailed description of the invention

It is highly surprising that the inventors of the present invention have found that 2- hydroxyoleic acid and derivatives thereof, such as compounds of formula (I), and pet food supplement comprising these compounds can be used for controlling the body weight of a pet selected from the group of a dog and a cat, such as an obese dog and an obese cat.

A first aspect of the invention relates to a compound of the general formula (I): COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

- m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

for use in the treatment of a pet selected from the group consisting of a dog and a cat. In a particular embodiment of the first aspect, the invention relates to 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof, for use in body weight management of an obese subject selected from the group consisting of an obese dog and an obese cat. A second aspect of the invention relates to a pet food supplement for feeding a pet, said pet food supplement comprising a compound of the general formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

m and n are independently selected from an integer of 0 to 15; and

- the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

A third aspect of the invention relates to pet food for feeding a pet, said pet food comprising a compound of the general formula (I): COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

- m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

where the compound of formula (I) is used as a pet food additive for controlling body weight of a pet selected from the group consisting of a dog and a cat. In a particular embodiment of the third aspect, the invention relates to a pet food for feeding an obese subject, said pet food comprising 2-hydroxyoleic acid or veterinary acceptable salt, ester or glyceride thereof, where said 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof is used as a pet food additive for body weight management of an obese subject selected from the group consisting of an obese dog and an obese cat.

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable, ester or glyceride salt thereof,

and where the pet is selected from the group consisting of a dog and a cat.

In a particular embodiment of the forth aspect, the present invention relates to a method of body weight management of an obese subject, comprising administering to said obese subject an effective amount of 2-hydroxyoleic acid or a veterinary acceptable salt, ester or glyceride thereof, where the obese subject is selected from the group consisting of an obese dog and an obese cat.

According to the compound of general formula (I), R may be selected from any group with a molecular weight from 14 to 200 Da, but preferably R is selected from H, OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN or N0₂. In a preferred embodiment of the compound of formula (I), R is H, OH, amino group, e.g. NH₂, or Ci-6 alkyl, e.g. CH₃. In one embodiment R is not H.

According to the compound of general formula (I), R may be selected from any group with a molecular weight from 14 to 200 Da, but preferably R is selected from OH, amino, d_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN or N0₂. In a preferred embodiment of the compound of formula (I), R is OH, amino group, e.g. NH₂, or C1-6 alkyl, e.g. CH₃.

In a most preferred embodiment of the compound of formula (I), R is selected from the group consisting of OH, an amino group, a hydrogen atom or Ci-₆ alkyl. In one specific embodiment R is OH. In another specific embodiment R is an amino group, e.g. NH₂,

In one embodiment, R is selected from Ci_-6 alkyl and C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy and hydrogen, such as wherein R is selected from Ci_-6 alkyl and C_2-6 alkenyl, C_2- 6 alkynyl or hydrogen. In a specific embodiment thereof R is hydrogen. In another specific embodiment R is not hydrogen.

In another embodiment, R is selected from Ci_-6 alkyl and C_2-6 alkenyl, C_2-6 alkynyl. In a specific embodiment thereof R is Ci_-6 alkyl, e.g. CH₃. In another specific embodiment thereof R is C_2-6 alkenyl. In yet another specific embodiment thereof R is C_2-6 alkynyl.

In another embodiment, R is selected from F, CI, CF₃, CCI₃, CN or N0₂.

In a specific embodiment, R is an amino group. In another specific embodiment, R is not an amino group.

In one preferred embodiment R may be selected from OH, NH₂, methoxy, F, CI, CF₃ and CCI₃. In another embodiment R may be selected from OH, methoxy, ethoxy, propoxy and isopropoxy, preferably OH or methoxy. In yet another embodiment R may be selected from OH, methyl, ethyl, propyl, isopropyl, ethenyl, propenyl, ethynyl and propynyl, preferably OH and methyl. In another embodiment R may be selected from OH, NH₂, methyl, methoxy and CF₃.

In one embodiment of the invention, m is selected from an integer of 6 to 14. More preferred, m is selected from an integer of 5 to 13, such as an integer from 6 to 12.

In one specific embodiment, m is selected from an integer of 5 to 9, such as an integer from 6 to 8. In another specific embodiment, m is selected from an integer of 10 to 14, such as an integer from 1 1 to 13. In one preferred embodiment, m is 6. In another preferred embodiment, m is 8. In yet another preferred embodiment, m is 12. In one embodiment of the invention, n is selected from an integer of 3 to 9, such as an integer of 4 to 8. In a specific embodiment, n is selected from an integer of 5 to 7. In a preferred embodiment, n is 5. In another preferred embodiment, n is 7. In a preferred embodiment of the invention the compound of the general formula (I) is 2-hydroxyoleic acid, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is 2-methyl-oleic acid, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is 2-amino-oleic acid, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is oleic acid, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is palmitoleic acid, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is cis-vaccenic acid, or a veterinary acceptable base salt thereof. In a further embodiment of the invention the compound of the general formula (I) is nervonic acid, or a veterinary acceptable base salt thereof. Examples of typical base salts include lithium, sodium, potassium, and magnesium salts or as elsewhere defined herein.

In yet a further embodiment of the invention the compound of general formula (I) is an ester compound, where 2-hydroxyoleic acid or the compound of general formula (I) has been reacted with methanol, ethanol, n-propanol, isopropanol or glycerol. In a preferred embodiment the ester compound has been formed by a reaction between glycerol and at least one fatty acid of general formula (I). In a particular preferred embodiment the ester compound has been formed by reacting glycerol with at least one 2-hydroxyoleic acid. In an especially preferred embodiment the compound is formed by reacting one glycerol molecule with one, two or three 2-hydroxyoleic acid molecules so as to form a monoglyceride, a diglyceride, or a triglyceride molecule.

In a preferred embodiment of the compound of general formula (I), the double bond is in cis configuration.

Salts of the invention Veterinary acceptable salts of a compound with an acid group include the anion of said compound in combination with a suitable cation such as sodium or ammonium ion. Other examples of veterinary acceptable salts include metal salts, ammonium and alkylated ammonium salts, salts of inorganic acids as well as organic acids.

Representative examples of suitable inorganic acids include hydrochloric, hydrobromic, hydroiodic, phosphoric, sulfuric, nitric acids and the like.

Representative examples of suitable organic acids include formic, acetic,

trichloroacetic, trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric, glycolic, lactic, maleic, malic, malonic, mandelic, oxalic, picric, pyruvic, salicylic, succinic, methanesulfonic, ethanesulfonic, tartaric, ascorbic, pamoic, bismethylene salicylic, ethanedisulfonic, gluconic, citraconic, aspartic, stearic, palmitic, EDTA, glycolic, p- aminobenzoic, glutamic, benzenesulfonic, p-toluenesulfonic acids and the like.

Examples of metal salts include lithium, sodium, potassium, magnesium salts and the like.

Examples of ammonium and alkylated ammonium salts include ammonium, methylammonium, dimethylammonium, trimethylammonium, ethylammonium, hydroxyethylammonium, diethylammonium, butylammonium, tetramethylammonium salts and the like.

Pets to be treated

The invention concerns treatment of a pet. The pet may be non-obese or the pet may be obese. In a particular embodiment, the pet to be treated is a dog or a cat. In one embodiment the pet is a non-obese dog. In one embodiment the pet is an obese dog. In one embodiment the pet is a non-obese cat. In one embodiment the pet is an obese cat.

Weight control

In some embodiments of the present invention the compounds of the invention, or veterinary acceptable salts, esters or glycerides thereof are for use as a pet food additive for controlling body weight of a pet, such as a dog or a cat. In one embodiment of the invention, the pet food additive is fed to a non-obese pet, such as a non-obese cat or a non-obese dog, whereby maintenance of weight is obtained. In one embodiment of the invention, the pet food additive is fed to an obese pet, such as an obese cat or an obese dog, whereby reduction of weight is obtained.

Pet food additive

In some embodiments the compounds of the invention are used as a pet food additive for controlling the body weight of a pet preferably selected from the group consisting of a dog and a cat, such as an obese dog and an obese cat. In some embodiments the pet food additive is 2-hydroxyoleic acid. The pet food additives may be mixed directly with the pet food, but preferably the pet food additives are incorporated into a separate pet food supplement, which comprises the pet food additives and optionally further excipients. By producing the separate pet food supplement it is possible to vary the amount of pet food additive, which must be fed to a particular pet. This is in particular beneficial for pet owners having two or more pets having different needs for the pet food additive. This could be if one pet is obese and the other pet is non-obese. In such case, both pets can be fed with the same pet food, whereas the amount of food additive can be easily varied by varying the amount of food supplement that is mixed with the pet food.

The pet food additive may be present in a pet food supplement, which is mixed with the pet food before a meal, or the pet food additive may be present in the pet food added by the manufacturer as part of the manufacturing process. The pet food additive may also be mixed with the drinking water, which is given to the pet. The pet food additive may be given to the pet with every meal or it may be given at a lower frequency, such as for example once a week or four times a week. Pet food supplement

The pet food supplement of the present invention comprises the pet food additive and optionally other ingredients. In some embodiments the pet food supplement only comprise 2-hydroxyoleic acid or one compound having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof, whereas in other embodiments the pet food supplement comprises a mixture of two or more different compounds being 2-hydroxyoleic acid and/or compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof.

The amount of pet food additive present in the pet food supplement typically lies in the range of 20-100%, such as for example 50-100%, more preferred in the range of 80- 100%. Examples of other excipients include fatty acids, which are not disclosed by general formula (I), minerals and vitamins.

The pet food supplement may be mixed with the pet food before a meal, or the pet food supplement may be mixed with the pet's drinking water.

Pet food

The pet food of the present invention is directed to any conventional pet food available on the market, which is produced for the purpose of feeding it to a pet as a meal, and which additionally comprises the pet food additive according to the present invention. A pet food of the present invention may be both a dry pet food and a wet pet food, which has been added the pet food additive according to the present invention. Accordingly, the pet food according to the present invention comprises, as a pet food additive, 2- hydroxyoleic acid or a compound having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof. It lies within the scope of the present invention that the pet food of the invention may also comprise a mixture of two or more different compounds being 2-hydroxyoleic acid and/or compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof. Pharmaceutical composition

The compounds of the invention, or veterinary acceptable salts, esters or glycerides thereof, to be used in accordance with the invention can be administered orally, in the form of tablets, capsules (including soft gel capsules), ovules, elixirs, solutions or suspensions, which may optionally contain flavouring or colouring agents.

In some embodiment the pharmaceutical compositions of the present invention comprises 2-hydroxyoleic acid or one compound having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof, whereas in other embodiments the pet food supplement comprises a mixture of two or more different compounds being 2-hydroxyoleic acid and/or compounds having the general formula (I) or a veterinary acceptable salt, ester or glyceride thereof.

In some embodiments the pharmaceutical composition may also comprise other active agents for weight control. An example of an active agent includes Dirlotapide, which is the active agent in the drug Slentrol.

Dosage

The effective amount of the pet food additive, which must be administered to a particular pet, depends on the race of the pet and on the pet's digestion and level of activity. The amount of pet food also depends on the weight of the pet. Furthermore, the amount depends on whether the pet is non-obese or obese. In one embodiment the pet food additive is administered to the pet as a daily dosage in a range of from about 1 to about 400 mg pet food additive / kg body weight, more preferred from about 10 to about 300 mg pet food additive / kg body weight, even more preferred from about 20 to about 200 mg pet food additive / kg body weight.

In one embodiment the pet food additive is administered to the pet as a daily dosage in a range of from about 60 to about 400 mg pet food additive / kg body weight, more preferred from about 100 to about 300 mg pet food additive / kg body weight, even more preferred from about 140 to about 260 mg pet food additive / kg body weight.

In one embodiment the pet food additive is administered to the pet as a daily dosage in a range of from about 160 to about 240 mg pet food additive / kg body weight, such as from about 170 to about 230 mg pet food additive / kg body weight, more preferred from about 180 to about 220 mg pet food additive / kg body weight, such as from about 190 to about 210 mg pet food additive / kg body weight, preferably 200 mg pet food additive / kg body weight. In another embodiment the pet food additive is administered to the pet as a daily dosage in a range of from about 160 to about 180 mg pet food additive / kg body weight, or from about 180 to about 200 mg pet food additive / kg body weight, or from about 200 to about 220 mg pet food additive / kg body weight, or from about 220 to about 240 mg pet food additive / kg body weight. Examples

The examples disclosed herein serve to test whether 2-hydroxyoleic acid (2-OHOA) have an effect on controlling body weight and on whether cats can tolerate the administration of 2-OHOA. The experiments were deigned to administer 2-hydroxyoleic acid in parallel with food intake in order to mimic the real usage situation as much as possible.

Example 1.

Weight loss study of 2-OHOA following oral administration in dogs.

Test animals

Species, Strain: Dog, Beagle

Source: Marshall Farms

Diet: Eukanuba High Performance

Number of Animals: Groups 1 -3: n=7 per group and Group 4: n=6 per group

Treatment regimen

Route, Dose Volume: Oral, 0.3ml/kg

Treatment Duration: Groups 1 & 2: 64 days; Group 3: 50 days; Group 4: 49 days

Treatment Groups:

Group 1 : Vehicle

Group 2: 2-OHOA; 50mg/kg BID (changed to 100mg/kg BID on day 20)

Group 3: 2-OHOA; 75mg/kg BID (changed to 150mg/kg BID on day 20)

Group 4: Sibutramine; 3mg/kg QD (changed to 5mg/kg QD on day 20)

Test and vehicle control articles

Test article:

2-OHOA (2-hydroxyoleic acid) - G2HOA-15

Sibutramine (positive control) - L28064

Vehicle:

2-OHOA - no vehicle, dosed in capsule in powder form

Sibutramine - 10% Acacia Results and discussion

Diet induced obese (DIO) dogs were randomized into four groups based upon body weight (i.e. percent change of initial body weight), Body Composition Scores (BCS), percent of fat mass and leptin values. The groups are as follows:

- Vehicle group (n=7): dogs were dosed for 64 days.

2-OHOA-low dose (n=7): Day 0, dogs were dosed with 50 mg/kg bid and then on day 20 switched to 100 mg/kg, bid. Dogs were dosed with 100 mg/kg, bid from day 20 to day 64.

- 2-OHOA-high dose (n=7): On day 0, dogs were dosed with 75 mg/kg bid and then on day 20 switched to 150 mg/kg bid. Dogs were dosed with the same dose until day 50.

2-Sibutramine group (n=6): On day 0, dogs were dosed at 3 mg/kg, qd for the first 20 days and then on day 20 they were switched to 5 mg/kg qd until day 49.

DIO dogs tolerated low doses of 2-OHOA and Sibutramine for the first 20 days and therefore the dose concentrations of both 2-OHOA and Sibutramine were increased for the rest of the experiment. All four groups were treated for 49 days, except for the 2- OHOA-high, which was treated for 50 days. Vehicle group and 2-OHOA-low group were treated for an additional 15 days after the initial 49 day treatment period.

Effect of oral administration of 2-OHOA on body weight in DIO dogs:

DIO dogs weighed between 10.00 - 1 1.19 kg at the initiation of treatment. Body weight data are presented as the change in body weight in the below Table and in Figure 1 . These data are the raw data, which have not been corrected by excluding outliers, i.e. test animals that did not respond to the treatment of 2-OHOA or Sibutramine, respectively. Hence, the below Table lists the average body weight loss of all tested dogs in each group. The body weight loss for a dog is calculated as the dog's body weight on a particular day minus the dog's initial body weight. A positive value is indicative for weight gain, whereas a negative value is indicative for weight loss.

Compared to vehicle-treated DIO dogs, 2-OHOA-low showed significant loss of body weight. The effect on loss of body weight was not as pronounced for the 2-OHOA -high treated group as compared with the 2-OHOA-low treated group. 2- 2- 2- 2-

OHOA OHOA Sibu- OHOA OHOA Sibu-

Day Vehicle Low High tramine Day Vehicle Low High tramine

1 0 0 0 0 33 0.1 1 -0.05 0.15 -0.50

2 -0.05 -0,05 -0.05 -0.23 34 0.1 1 -0.10 0.01 -0.40

3 0.08 0 0 -0.17 35 0 -0.17 -0.06 -0.55

4 0.02 -0.03 0.05 -0.13 36 0.05 -0.17 -0.08 -0.47

5 0.02 0.06 0.08 -0.13 37 -0.03 -0.23 -0.05 -0.52

6 0.15 0.12 0.08 -0.20 38 0.05 -0.18 -0.13 -0.58

7 0.08 0.12 0.10 -0.07 39 -0.02 -0.20 -0.13 -0.62

8 0.1 1 0.03 0.05 -0.15 40 0.02 -0.18 -0.13 -0.55

9 0.08 0.12 0.12 -0.15 41 0.02 -0.08 -0.06 -0.50

10 0.15 0.05 0.10 -0.20 42 0.08 -0.13 -0.08 -0.47

1 1 0.10 0.10 0.12 -0.13 43 0.04 -0.10 -0.16 -0.43

12 0.12 0 0.08 -0.17 44 -0.02 -0.24 -0.09 -0.42

13 0.15 0.07 0.17 -0.08 45 0.02 -0.23 -0.05 -0.48

14 0.01 -0.07 0.05 -0.18 46 0.01 -0.18 -0.06 -0.43

15 0.01 -0.01 0.01 -0.18 47 -0.05 -0.24 -0.16 -0.50

16 0.04 -0.03 0.07 -0.22 48 -0.02 -0.14 -0.09 -0.43

17 0.08 -0.10 0.1 1 -0.32 49 -0.19 -0.37 -0.28 -0.65

18 0.1 1 -0.03 0.20 -0.20 50 -0.02 -0.28 -0.12 -0.45

19 0.18 0.02 0.12 -0.23 51 -0.08 -0.30 -0.03

20 0.14 0.09 0.18 -0.20 52 -0.16 -0.33

21 0.08 -0.04 0.18 -0.38 53 0.02 -0.14

22 0.07 -0.04 0.15 -0.40 54 -0.02 -0.21

23 0.05 -0.01 0.12 -0.42 55 0.01 -0.18

24 0.10 -0.01 0.1 1 -0.50 56 -0.05 -0.30

25 0 0.05 0.14 -0.45 57 -0.09 -0.35

26 0.21 0.02 0.10 -0.53 58 0.02 -0.25

27 0.1 1 0.12 0.25 -0.45 59 -0.08 -0.27

28 0.07 -0.08 0.02 -0.52 60 -0.03 -0.25

29 0 -0.15 -0.02 -0.55 61 0.08 -0.21

30 0.02 -0.13 -0.05 -0.53 62 0.02 -0.1 1

31 0.08 -0.10 0.02 -0.52 63 0.07 -0.21

32 0.07 -0.1 1 0 -0.53 64 0.1 1 -0.30

The most effective dose of 2-OHOA (i.e. 2-OHOA-low, 50mg/kg followed by 100 mg/kg) was administered to DIO dogs for a prolonged period i.e. 64 days in order to determine i) abnormal clinical signs and ii) further loss of body weight. DIO dogs treated with 2-OHOA did not show any abnormal clinical signs or symptoms. They showed a consistent level of body weight loss over the dosing period.

Effect of oral administration of 2-OHOA on cumulative food intake:

DIO dogs treated with both dose levels of 2-OHOA showed similar food intake compared to the vehicle treated group. On the other hand, Sibutramine-treated dogs showed a significant decrease in food intake, more prominent after 21 days of treatment. This was expected as this is the mode of action of Sibutramine and thereby it confirms the validity of the trial design and model.

Example 2.

Dose range finding study of 2 -OH OA following oral administration in cats.

Animals

The cats were supplied by Liberty Research, Inc. Documentation included the animal number, ear tattoo and date of birth. At arrival all animals were visually examined for external signs of illness. No further treatment was carried out during the acclimatization period. Prior to the first dosing, relevant clinical health records, visual examination, body weight, food consumption data and clinical pathology data were reviewed by the study director to ensure that the animals were in excellent or very good general condition. Only healthy animals were included in the study. Species / Strain: Cat / Domestic short-hair feline

Breeder: Liberty Research, Inc., 170 State Route 17C, Waverly, NY 14892, USA Number of animals: 4 neutered male animals

Age (at first administration): Approx. 3 years

Body weight (at first administration): 5.0 kg, 4.7 kg, 4.2 kg and 4.8 kg

Identification of animals: By ear tattoo.

General adaptation period: 24 days

Adaptation period to single housing (metabolism cages): 7 days

Preparation of the test item solution

The test item-vehicle solution was freshly prepared on each administration day according to the following formulation instructions:

The required amount of the test item was weighed into a mortar and ground. The total amount of glycerol to prepare the complete formulation needed for dosing was added and mixed with the test item. Distilled water (not the whole amount needed for the formulation) was added drop-wise whilst mixing.

The resulting solution was transferred to a volumetric flask and was stirred slowly on a magnetic stirrer until a homogenous suspension was obtained. After the foam had fully subsided, the suspension was stored at room temperature protected from light. The flask was filled with distilled water to the total volume needed for administration and the suspension was stirred slowly until complete homogenisation.

Administration

Route of administration: Oral, by gavage.

Frequency of administration: Repeated administration for 3 days per dosing cycle. Vehicle: 2% glycerol in distilled water

Administration volume: Cycles 1 and 2: 2 mL/kg b.w. and Cycles 3 and 4: 4 mL/kg b.w. The amount of the test item was adjusted to each animal's current body weight on the first day of each cycle.

Animals were fasted overnight (at least 12 hours) prior to each administration. The animals were fed 2 hours after the administration. The time of feeding and dosing was documented in the raw data.

Group size / dose levels

The four test animals were all subjected to 4 cycles of treatment. Two subsequent cycles of treatment were separated by a wash-out period:

Dose levels: Cycle 1 : 30 mg 2-OHOA/kg b.w./day

Cycle 2: 100 mg 2-OHOA/kg b.w./day

Cycle 3: 300 mg 2-OHOA/kg b.w./day

Cycle 4: 200 mg 2-OHOA/kg b.w./day

Wash-out period between the cycles 1 , 2 and 3 was 72 h.

Wash-out period between the cycles 3 and 4 was 19 days.

The administration was repeated for 3 days per dosing cycle.

The treatment schedule can also be summarised according to the below table: Date of Number and

2-OHOA dose Frequency of

Group administration sex of

[mg/kg b.w.] administration

animals

30

1 3 days April 20, 2012 1 - 3 m

Dose 1

Wash-out period of 72 h

100

2 3 days April 25, 2012 1 - 3 m

Dose 2

Wash-out period of 72 h

300

3 3 days April 30, 2012 1 - 3 m

Dose 3

Wash-out period of 19 days

3 days May 21 , 2012 1 - 3 m^#

4## 200

Dose 4

3 days May 30, 2012 4 m m: male

# animal no. 1 died accidently during the 2nd application of cycle 4. Therefore, a fourth animal was included to this cycle and started dosing on May 30. ## a dose of 200 mg/kg b.w. was added as the side-effects observed at 300 mg/kg b.w., in particual vomiting, reduced food intake appeared to be too pronounced for a MTD.

Housing and feeding

Diet

Commercial ssniff^® Cat, V3146 Extrudat served as food. The food was offered to each cat once daily i.e. the animals had access to the food ad libitum. The remaining food was removed next morning; residues were discarded. Prior to the oral administration, animals were fasted over night. Feeding was resumed 2 hours post dosing.

Drinking water

Drinking water was offered ad libitum.

Housing during the pool/general adaptation period The cats were kept in groups; the housing temperature was maintained at a

temperature of 22°C ± 3°C (maximum range). Drinking water was available ad libitum. The diet was offered once daily. Housing during 7-day adaptation to individual housing and during the experimental period

The cats were kept singly in cages; the housing temperature was maintained at a temperature of 22°C ± 3°C (maximum range). Drinking water was available ad libitum. The diet was offered once daily.

Results

Mortality

No test item-related mortality was noted for the animals treated orally with 30, 100, 200 or 300 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day three days per cycle. However, animal no. 1 treated with 200 mg 2-OHOA(2-hydroxyoleic acid)/kg b.w./day died accidently during the 2nd application of cycle 4. Rarely, cats become very

uncooperative during dosing and the cat might have died due to aspiration of some test material. Hence, a fourth animal was included to this cycle, started dosing on May 30, 2012.

Behaviour, external appearance and faeces

None of the animals treated in the cycle 1 with 30 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day showed any clinical signs of systemic toxicity. Animal no. 1 treated in the cycle 2 with 100 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day vomited once 5 to 20 minutes after the first administration of cycle 2 (approx. 5 ml_). The remaining animals treated with 100 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day did not show any clinical signs of systemic toxicity. Treatment with 200 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day in the cycle 4 caused following changes: Animal no. 2 vomited once immediately after the second administration of the test item (approx. 5 mL) and animal no. 4 vomited once, 20 to 60 minutes after each administration of the test item (approx. 5 to 10 mL). Treatment with 300 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day in the cycle 3 caused following changes: Animal no. 1 vomited three times immediately after the first administration of cycle 3 (approx. 10 mL, 5 mL and 3 mL) and three times after the second administration of cycle 3 (approx. 10 mL and 5 mL). Animal no. 2 vomited once, 2 to 6 hours after the first administration of cycle 3, once 5 to 20 min after the second administration and twice 1 to 2 hours after the second administration of cycle 3

(approx. 10 mL). The remaining animal no. 3 treated with 300 mg 2-OHOA (2- hydroxyoleic acid)/kg b.w./day revealed no signs of systemic intolerance. Exposure

Dose/exposures in cat are similar to rats where significant anti-obesity/diabetes measures were obtained.

Body weight

The body weight was not influenced in the animals treated with 30, 100, 200 or 300 mg 2-OHOA(2-hydroxyoleic acid)/kg b.w./day compared to the predose value of the respective cycle. The aim of the study was not to show a weight impact but

demonstrate exposure levels obtained in other species whereby the effect results can be bridged.

Food and drinking water consumption

The food intake of the animals treated with 30, 100 or 200 mg 2-OHOA/kg b.w./day appeared to be in the normal range. A decrease of the food intake was noted for all animals treated with 300 mg 2-OHOA (2-hydroxyoleic acid)/kg b.w./day by up to 51 % compared to the predose value of the respective cycle.

The visual appraisal of drinking water consumption did not reveal any test item-related influence.

Discussion

Weight control is obtained by balancing energy intake with energy loss through the metabolism. In principle energy intake can be controlled/reduced by the following strategies: - appetite regulation/suppression (Sibutramine),

- energy absorption in the intestines (orlistat), or

- energy regulation (amphetamines). Previously, 2-hydroxyoleic acid studies in rodents have suggested appetite regulation as a mode of action - possibly in combination with energy regulation. Reduced energy absorption in the intestines of cats and dogs can be ruled out due to the lack of effect on the stool which is seen with orlistat. The present trial shows no impact on food consumption and the weight management effect therefore must be contributed to an energy regulation effect not yet elucidated.

Claims

1 . A compound of the general formula (I): COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

- m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

for use in the treatment of a pet selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

2. The compound or a veterinary acceptable salt, ester or glyceride according to claim 1 , wherein R in formula (I) is selected from H, OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

3. The compound according to any one of claims 1 to 2, wherein the compound of the general formula (I) is 2-hydroxyoleic acid, 2-methyl-oleic acid, 2-amino-oleic acid, oleic acid, palmitoleic acid, cis-vaccenic acid or nervonic acid or a veterinary acceptable base salt, ester or glyceride thereof.

4. The compound according to claim 1 , wherein

R represents a group selected from OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN, N0₂, or some other group with a molecular weight from 14 to 200 Da,

m and n are independently selected from an integer of 0 to 15; and

- the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

5. The compound or a veterinary acceptable salt, ester or glyceride according to claim 4, wherein R in formula (I) is selected from OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

6. The compound according to any one of claims 4 to 5, wherein the compound of the general formula (I) is 2-hydroxyoleic acid, 2-methyl-oleic acid or 2-amino-oleic acid or a veterinary acceptable base salt, ester or glyceride thereof.

7. The compound according to any of claims 4 to 6, wherein the compound is 2- hydroxyoleic acid or a veterinary acceptable base salt, ester or glyceride thereof.

8. The compound or an veterinary acceptable salt, ester or glyceride according to any one of claims 1 -7 for use as a pet food additive for controlling body weight, such as by maintaining body weight in a non-obese pet or by reducing body weight in an obese pet, where the pet is selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

9. The compound or a veterinary acceptable salt, ester or glyceride thereof according to any one of claims 1 -8, which is administered to the pet as a daily dosage in a range of from about 1 to about 400 mg / kg body weight, more preferred from about 10 to about 300 mg / kg body weight, even more preferred from about 20 to about 200 mg / kg body weight.

10. A pet food supplement for feeding a pet, said pet food supplement comprising a compound of the general formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof, where the compound of formula (I) is used as a pet food additive for controlling body weight of a pet selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

1 1. The pet food supplement according to claim 10, wherein R in formula (I) is selected from H, OH, an amino group, e.g. NH₂, or Ci_₆ alkyl, e.g. CH₃.

12. The pet food supplement according to any one of claims 10 to 1 1 , wherein the pet food supplement comprises one or more compound(s) of the general formula (I) selected from the group consisting of 2-hydroxyoleic acid, 2-methyl-oleic acid, 2-amino- oleic acid, oleic acid, palmitoleic acid, cis-vaccenic acid and nervonic acid and veterinary acceptable base salts, esters or glycerides thereof.

13. The pet food supplement according to claim 10 comprising a compound of the general formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

where the compound of formula (I) is used as a pet food additive for controlling body weight of a pet selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

14. The pet food supplement according to claim 13, wherein R in formula (I) is selected from OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

15. The pet food supplement according to any one of claims 13 to 14, wherein the pet food supplement comprises one or more compound(s) of the general formula (I) selected from the group consisting of 2-hydroxyoleic acid, 2-methyl-oleic acid and 2- amino-oleic acid and veterinary acceptable base salts, esters or glycerides thereof.

16. The pet food supplement according to any of claims 13 to 15, wherein the compound is 2-hydroxyoleic acid or a veterinary acceptable base salt, ester or glyceride thereof.

17. The pet food supplement according to any one of claims 10 to 16, wherein the pet food supplement is fed to a pet for controlling body weight, such as by maintaining body weight in a non-obese pet or by reducing body weight in an obese pet, where the pet is selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

18. The pet food supplement according to any one of claims 10 to 17, wherein the pet food supplement comprises at least one compound of general formula (I) as a pet food additive and optionally one or more further excipients.

19. The pet food supplement according to any of claims 10 to 18, which is fed to the pet as a daily dosage, is ranging from about 1 to about 400 mg compound of general formula (I) / kg body weight, more preferred from about 10 to about 300 mg compound of general formula (I) / kg body weight, even more preferred from about 20 to about 200 mg compound of general formula (I) / kg body weight.

20. A pet food for feeding a pet, said pet food comprising a compound of the general formula (I): COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

- m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

21. The pet food according to claim 20, wherein R in formula (I) is selected from H, OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

22. The pet food according to any one of claims 20 to 21 , wherein the pet food comprises one or more compound(s) of the general formula (I) selected from the group consisting of 2-hydroxyoleic acid, 2-methyl-oleic acid, 2-amino-oleic acid, oleic acid, palmitoleic acid, cis-vaccenic acid and nervonic acid and veterinary acceptable base salts, esters or glycerides thereof.

23. The pet food according to claim 20 comprising a compound of the general formula (I):

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

m and n are independently selected from an integer of 0 to 15; and

- the double bond is in cis or trans configuration,

or a veterinary acceptable salt, ester or glyceride thereof,

24. The pet food according to claim 23, wherein R in formula (I) is selected from OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

25. The pet food according to any one of claims 23 to 24, wherein the pet food comprises one or more compound(s) of the general formula (I) selected from the group consisting of 2-hydroxyoleic acid, 2-methyl-oleic acid and 2-amino-oleic acid and veterinary acceptable base salts, esters or glycerides thereof.

26. The pet food according to any of claims 23 to 25, wherein the compound is 2- hydroxyoleic acid or a veterinary acceptable base salt, ester or glyceride thereof.

27. The pet food according to any one of claims 20 to 26, wherein the pet food is fed to a pet for controlling body weight, such as by maintaining body weight in a non-obese pet or by reducing body weight in an obese pet, where the pet is selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

28. A method of controlling body weight of a pet, comprising administering to a pet an effective amount of a compound of formula (I)

COOH-CHR-(CH₂)_m-CH=CH-(CH₂)_n-CH₃ (I), wherein

m and n are independently selected from an integer of 0 to 15; and

the double bond is in cis or trans configuration,

or a veterinary acceptable, ester or glyceride salt thereof,

and where the pet is selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

29. The method according to claim 28, wherein R in formula (I) is selected from H, OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

30. The method according to any one of claims 28 to 29, wherein the compound of the general formula (I) is 2-hydroxyoleic acid, 2-methyl-oleic acid, 2-amino-oleic acid, oleic acid, palmitoleic acid, cis-vaccenic acid or nervonic acid or a veterinary acceptable base salt, ester or glyceride thereof.

31. The method according to claim 28, wherein

R represents a group selected from OH, amino, Ci_-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, Ci_-6 alkoxy, F, CI, CF₃, CCI₃, CN, N0₂, or some other group with a molecular weight from 14 to 200 Da

m and n are independently selected from an integer of 0 to 15; and

- the double bond is in cis or trans configuration, or a veterinary acceptable, ester or glyceride salt thereof,

32. The method according to claim 31 , wherein R in formula (I) is selected from OH, an amino group, e.g. NH₂, or Ci_-6 alkyl, e.g. CH₃.

33. The method according to any one of claims 31 to 32, wherein the compound of the general formula (I) is 2-hydroxyoleic acid, 2-methyl-oleic acid and 2-amino-oleic acid or nervonic acid or a veterinary acceptable base salt, ester or glyceride thereof.

34. The method according to any of claims 31 to 33, wherein the compound is 2- hydroxyoleic acid or a veterinary acceptable base salt, ester or glyceride thereof.

35. The method according to any one of claims 28 to 34, wherein the compound of the general formula (I) is administered to a pet for controlling body weight, such as by maintaining body weight in a non-obese pet or by reducing body weight in an obese pet, where the pet is selected from the group consisting of an obese dog, an obese cat, a non-obese dog and a non-obese cat.

36. The method according to any one of claims 28 to 35, wherein the compound of the general formula (I) is administered to the pet as a daily dosage in a range of from about 1 to about 400 mg / kg body weight, more preferred from about 10 to about 300 mg / kg body weight, even more preferred from about 20 to about 200 mg / kg body weight.