EDIBLE FOAM COMPRISING A DRUG

Introduction

The present invention relates to a foamed composition for use in the treatment of administering a drug or mixtures of drugs to an individual. The invention also relates to a method of ingesting a drug or mixture of drugs in the form of or as part of an edible foamed composition.

Background of the invention

By far, the most common route of administering a pharmacologically active component (such as e.g. medicaments or drugs having a specific function) to an individual is by oral ingestion. Such oral ingestion is commonly in the form of tablets, capsules, drinks, powders and the like. However, oral ingestion of a pharmacologically active component in any of the

aforementioned conventional formats is no guarantee to desired effect. After oral ingestion, bioavailability of such pharmaceutically active compound e.g. depends on proper release from the chosen format and solubility of said compound as well as the permeability of said compound. Taking this in mind, intestinal drug absorption can be classified in the so-called BCS system (Biopharmaceutics Classification System, as provided by the US Food and Drug Administration, and as further referred to by Amidon, G.L., Lennernas, H., Shah, V.P. et al in: A theoretical basis for a biopharmaceutic drug classification - the correlation of in-vitro drug product dissolution and in-vivo bioavailability; Pharmaceutical Research; 1995, Vol. 12, no.3, p. 413-420; and in Guidance for Industry: Waiver of In Vivo Bioavailability and Bioequivalence Studies for Immediate-Release Solid Oral Dosage Forms Based on a Biopharmaceutics Classification System, US Dept of Health and Human Services, FDA, 2000; and by Lobenberg, R., Amidon, G.L.; Modern bioavailability, bioequivalence and biopharmaceutics classification system. New scientific approaches to international regulatory standards; European Journal of Pharmaceutics and Biopharmaceutics; 2000, Vol. 50, no.1 , p. 3-12). Following this classification, drugs for use by humans can be classified as BCS Class I, Class II, Class III and Class IV drugs.

BCS Class I drugs are characterized by a high solubility in combination with a high permeation rate of such drugs across the intestinal wall. This typically results in fast absorption kinetics, resulting a quick peak plasma level (i.e. a short Tmax: time at which the peak plasma concentration is observed) and high concentration at Tmax (i.e. a high Cmax: high peak plasma value).

BCS Class II drugs are characterized by a low solubility in combination with a high permeation rate of such drugs across the intestinal wall.

BCS Class III drugs are characterised by a rapid dissolution and low permeation across the intestinal wall. BCS Class IV drugs are characterised by a low dissolution and low permeation across the intestinal wall.

Usually, pharmacologically active components such as drugs are in the form of small solid dosings, such as e.g. pills, tablets or capsules. Oral intake of such small solid dosings is most commonly (physically) facilitated by ingestion of a certain amount of liquid, e.g.

water. This is also how consumers are used to such. The volume of such liquid that is ingested by the consumers can differ widely among individuals: from a mouthful to a glass, such being dependent on instructions for use but also on habits, likings or beliefs by such individual consumers.

A problem encountered in the pharmacological field is that of variability in

pharmacokinetics. Most normal solid oral dose formats (capsules, tablets, pills) of drugs show a wide variability with respect to absorption of the drug, due to large differences in stomach-intestine motility and differences in time to pass from stomach to intestine. These differences are the result of two effects: firstly of all the inter-individual differences in longitudinal transport rate in humans in the small intestine, and secondly also due to differences in transport rate within the same subject due to the different phases of the migrating motor complex the subject is in while taking the oral dose. This variability in the pharmacokinetics among individuals is disadvantageous, and worsened by differences in intake with or without liquid, amount of such, and whether or not the drug is consumed just before, with just after, or some time after ingestion of (solid) food. Differences in PK (pharmacokinetic) variability leads to a wide variability in the plasma concentration versus time profile (PK profile): plasma concentrations become unpredictable and hence desired effects and unwanted side effects are also unpredictable. Differences in PK profile could relate to the onset of action or differences in plasma concentrations reached. Avoidance of lag-time effects is especially desired where the onset of action should be fast and absorption be unrestricted (e.g. painkillers, drugs used for spasm and seizures, drugs with stimulating effects). Differences in plasma concentration reached are undesired for drugs with a narrow therapeutic window (warfarin, a BCS class I drug, most anti-cancer drugs, lithium, a BCS Class I drug, phenytoine, a BCS Class II drug) or drugs which have a minimal effective concentration (antibiotics such as doxycycline, a BCS Class I drug).

Apart from variability in pharmacokinetics it is known that some drugs are best taken with food (preferably a controlled amount or composition of food). Examples of drugs to be taken with food are all those which give local irritation on the Gl wall (e.g. iron salts, salicylic acid derivatives, and many BCS Class II drugs), or depend on the solubility- enhancing effects of Gl juices (most BCS class II drugs). Despite that such drugs are usually accompanied with proper instructions for use, consumers may find it difficult to adhere such instructions. It is both desired that the variability in pharmacokinetics is reduced between different subjects, but also within a subject over multiple events the drug is ingested. Hence, reduced variability as a target here covers both.

Next to that, there are also drugs that are preferably taken with no food or only a controlled amount, such as for example drugs which should have fast onset of action, all drugs which are vulnerable to gastric acid or pancreatic enzymes (for example but not limited to esters, amides and those with a peptide bond), drugs complexing with common food components such as iron and calcium (e.g. tetracyclines, bisphosphonates), drugs of which the absorption is decreased or delayed such as most antibiotics, beta-blockers, cyclosporine, etc. Drugs not to be taken with acid beverages are e.g. ketoconazole, etc. Source: Drug Induced Nutrient Depletion Handbook, American Pharmaceutical

Association, 1999-2000.

GB 1 121358 discloses the use of an aerosol foam dosage for the administration of a cough remedy. US 5369131 discloses liquid pharmaceutical compositions which can be administered as a foam. Such foam is disclosed for oral, cutaneous, and intravaginal use.

EP 362655 discloses foam for oral ingestion, which foam contains high amounts (45 to 78 weight%) of cough syrup, cough drops concentrate, sennoside (senna) syrup, multivitamin syrup, valerian syrup. WO 2010/144865 discloses compositions for treating gastrointestinal disorders, especially but not exclusively of the esophagus. The compositions are for topical treatment of the gastrointestinal tract, as alternative to systemic treatment. The compositions comprise at least one therapeutic agent and are formulated to increase the interaction of the composition comprising said therapeutic agent. Foams are disclosed as an alternative for those having difficulty to swallowing a viscous liquid or solid oral dose.

WO 2008/009617 A1 and WO 2008/009623 A1 disclose foams that may contain a functional ingredient. These disclosures are silent about controlled and/or delayed uptake of drugs, and teach that other compounds may be added to the foam to delay or control the uptake of an active additive.

WO 2008/046729 A1 discloses foams to increase satiety of a consumer.

WO 2008/046699 A1 discloses foams in which the gas bubbles are stabilised by insoluble fibres assembled with surface-active particles. Both references are silent on the uptake of drugs from the foams.

Summary of the invention

Hence, there is a need for an alternative method or system for orally administering pharmacologically active compounds (e.g. drugs that belong to BCS Class I, II, III or IV, including mixtures of drugs, in particular for drugs that belong to BCS Class I and II, more in particular for drugs that belong to BCS Class I, and preferably such method should reduce variability in pharmacokinetics with regard to rate of absorption of such drugs in humans, when compared to conventional intake of drugs. Preferably pharmacokinetic (PK) variability should be reduced to the smallest possible level in a given experimental design to result in a small variability around the PK parameters which reflect the absorption rate and extent of absorption. More specifically the variability in rate of drug absorption (Ka) ("speed" at which a drug enters the central compartment) and/or of Tmax (the time that the highest plasma concentration is achieved) should be small, and preferably smaller than is achieved with existing dosing methods. Preferably, the ratio between the highest and lowest value of Ka when measured for the same drug in the same dosing in a group of test persons should be less than 10, more preferably less than 6, and preferably the ratio between the highest and lowest value of Tmax when measured for the same drug in the same dosing in a group of test persons should be less than 4, more preferably less than 3. ln addition or apart from this, there is a need for a method or system that not only provides the drug, but that also provide a degree of satiety, as such is believed to lower the risk that consumers will take drugs with food whilst such is undesired. In addition or apart from this, there is also a need for a method or system that not only provides the drug, but that provides a controlled amount and composition of food or drink, in such a way that there is a reduced tendency for users to consume more food and/or drink with such drugs.

It has now been found that the above objectives, may be achieved, at least in part, by a foamed composition for use in the treatment of administering a drug or mixtures of drugs to an individual, wherein the treatment comprises

orally ingesting by said individual

a dosing of 50 - 600 ml of a foamed composition,

and which foamed composition:

has a continuous aqueous liquid phase comprising at least 60% by weight of water,

has a plurality of gas bubbles dispersed in said continuous aqueous liquid phase, has an overrun of at least 50%,

comprises said drug or mixtures of drugs,

is characterised by a high in-mouth foam stability being evidenced by a reduction in overrun of less than 35% under in-mouth conditions, using the methodology defined in the description.

The invention further relates to a method for administering a drug or mixtures of drugs to an individual, the method comprising

- orally ingesting by said individual

a dosing of 50 - 600 ml of a foamed composition,

and which foamed composition:

has a continuous aqueous liquid phase comprising at least 60% by weight of water,

- has a plurality of gas bubbles dispersed in said continuous aqueous liquid phase, has an overrun of at least 50%,

comprises said drug or mixtures of drugs,

is characterised by a high in-mouth foam stability being evidenced by a reduction in overrun of less than 35% under in-mouth conditions, using the methodology defined in the description. The invention further relates to the use of a foamed composition, in the manufacture of a medicament comprising a drug or mixtures of drugs for a therapeutic or prophylactic treatment, which therapeutic or prophylactic treatment comprises

orally ingesting by said individual

- a dosing of 50 - 600 ml of a foamed composition,

and which foamed composition:

has a continuous aqueous liquid phase comprising at least 60% by weight of water,

has a plurality of gas bubbles dispersed in said continuous aqueous liquid phase, - has an overrun of at least 50%,

comprises said drug or mixtures of drugs,

is characterised by a high in-mouth foam stability being evidenced by a reduction in overrun of less than 35% under in-mouth conditions, using the methodology defined in the description.

The invention further relates to the use of a foamed composition comprising a drug or mixtures of drugs for reducing the variability in the PK profile of said drug, by orally ingesting a dosing of 50 - 600 ml of a foamed composition, and which foamed composition:

- has a continuous aqueous liquid phase comprising at least 60% by weight of water,

has a plurality of gas bubbles dispersed in said continuous aqueous liquid phase, has an overrun of at least 50%,

comprises said drug or mixtures of drugs, and

- wherein the foamed composition is characterised by a high in-mouth foam stability being evidenced by a reduction in overrun of less than 35% under in-mouth conditions, using the methodology defined in the description.

The composition, method and use according to the present invention are particularly advantageous for drugs which have a high permeation rate cross the intestinal wall, and which are either easily dissolvable or less well soluble, for reasons given above.

Limited variability in plasma concentration profile, expressed in low variability around the PK parameters, such as rate of absorption (Ka) and time to reach the maximum concentration in plasma (Tmax), is in general a desirable aspect of drugs and drug formulations. This is as plasma concentration versus time profile relates to onset and extent of effect and side effects. This is in particular valid for those drugs which are not limited in their solubility in the dosage form or gastro-intestinal fluid, or where the rate of absorption is slow and controlled by the gut wall. The absorption of such drugs, which are highly soluble and not limited in gut wall absorption is to a large extent or in total controlled by gastric emptying. These drugs are classified as BCS Class I drugs. For this reason, it is preferred that in the composition, method and use as specified herein that drug is a drug of BCS Class I, BCS Class II, BCS Class III or BCS Class IV, preferably of BCS Class I or of Class II, most preferably of BCS Class I. It is preferred (e.g. to distinguish it from food eaten for pleasure) that in the composition, method and use as specified above the drug or mixtures of drugs thereof is a

pharmacologically active compound, and even more preferably that it is present in an effective amount. Detailed description

"Drug" herein comprises the chemical compound as such, as well as its commonly used forms such as its salt(s), oxide, hydrate, complex, ester, or other pharmacologically active form. "BCS Class I drug" is herein to be understood as all components in the group of Class I as defined by the US Food and Drug Administration in the Biopharmaceutics Classification System. The group of drugs which are BCS Class I drugs herein comprises, but is not limited to:

abacavir, acetaminophen (= paracetamol), acyclovirb, amiloride, amitryptyline, antipyrine, atropine, buspironec, caffeine, captopril, chloroquine, chlorpheniramine,

cyclophosphamide, desipramine, diazepam, diltiazem, diphenhydramine, disopyramide, doxepin, doxycycline, enalapril, ephedrine, ergonovine, ethambutol, ethinyl estradiol, fluoxetine, imipramine, ketorolac, ketoprofen, labetolol, levodopajevofloxacin, lidocaine, lomefloxacin, meperidine, metoprolol, metronidazole, midazolam, minocycline, misoprostol, nifedipine, phenobarbital, phenylalanine, prednisolone, primaquine, promazine, propranolol, quinidine, rosiglitazone, salicylic acid, theophylline, valproic acid, verapamil, zidovudine, drugs not named herein but classified as BCS Class I drug, and mixtures thereof. "BCS Class II drug" is herein to be understood as all components in the group of Class II as defined by the US Food and Drug Administration in the Biopharmaceutics Classification System. The group of drugs which are BCS Class II drugs herein comprises, but is not limited to:

amiodarone, atorvastatin, azithromycin, carbamazepine, carvedilol, chlorpromazine, cisapride, ciprofloxacin, cyclosporine, danazol, dapsone, diclofenac, diflunisal, digoxin, erythromycin, flurbiprofen, glipizide, glyburide, griseofulvin, ibuprofen, indinavir, indomethacin, itraconazole, ketoconazole, lansoprazole, lovastatin, mebendazole, naproxen, nelfinavir, ofloxacin, oxaprozin, phenazopyridine, phenytoin, piroxicam, raloxifene, ritonavir, saquinavir, sirolimus, spironolactone, tacrolimus, talinolol, tamoxifen, terfenadine, drugs not named herein but classified as BCS Class II drug, and mixtures thereof.

"Pharmacologically active compound" is herein to be understood as defined by the US Food and Drug Administration as "drug": (A) articles recognized in the official United States Pharmacopoeia, official Homoeopathic Pharmacopoeia of the United States, or official National Formulary, or any supplement to any of them; and (B) articles intended for use in the diagnosis, cure, mitigation, treatment, or prevention of disease in man or other animals; and (C) articles (other than food) intended to affect the structure or any function of the body of man or other animals; and (D) articles intended for use as a component of any article specified in clause (A), (B), or (C).

"Effective amount" herein is the amount per dosing that gives a therapeutic effect (for the drug stated), when used in an accepted treatment based on the pharmacologically active compound. "Foam", "foamed composition", "foam product" and "aerated composition" are herein used interchangeably: they mean herein the same. "Aerated" does not imply that the gas bubbles dispersed in the continuous liquid phase is air or has the same composition as air. An aerated composition, like a foam, foam product or foamed composition herein, can have any gas as specified herein dispersed as bubbles in the continuous liquid phase. Aerated or foamed means the presence of a compound in gas phase at temperatures of between 1 and 40°C and atmospheric pressure, wherein the gas can be air but also other gaseous compounds (i.e. aerated can be with other cases than air). "Overrun" of a foamed product is calculated using the following equation:

Overrun = 100% x (V _{foam product} - V _mix ) / V _mix foam roduct = Volume of a sample of the edible foamed product

V _mix = Volume of the same sample after the dispersed gas phase has been removed.

It was found that foamed systems which are (without containing a drug) known to be able to induce satiety after consumption) can be combined with a drug or mixtures of drugs, which combination can be beneficial for drugs which are preferably not combined with food of substance. This applies to food taken some time before or some time after the drug is ingested. It is believed that the satiety induced by a sufficient amount of foam reduced the tendency of individuals to consume food relatively shortly after consumption of the drug. For this purpose, a minimum amount of foam is required: at least 50 ml, preferably at least 75 ml, more preferably at least 100 ml, and the foam should have sufficient stability as expressed as a high in-mouth foam stability being evidenced by a reduction in overrun of less than 35% under in-mouth conditions, using the methodology defined herein below.

Additionally, it was found that a sufficient amount of foam as described herein (the same amounts and foam stability as described above with regard to satiety), when used as a vehicle for a drug or mixture of drugs, can give such drug a reduced variability in pharmacokinetics. Stomach emptying was equally quick (for foam according to the invention as for liquid of the same weight) as demonstrated by virtually identical average GE50 values (halftime of Gastric Emptying, i.e. the time required to reach half of the foam emptied from the stomach). Surprisingly (as with unchanged GE50 for foams and liquids of the same weight) however, a considerable difference in variability of Ka and Tmax was obtained. What was found was a narrower range of rate of absorption (Ka) values for the foam (experimental herein: 13 - 43 min ^"1 , a ratio of 3.36) versus the liquid (7-92 min ^"1 , a ratio of 13.02), and Tmax values for the foam (experimental herein: 47-102 minutes: a ratio of 2.2) versus the liquid (28-133 minutes: a ratio of 4.7). Obtaining reduced variability in PK profile (as expressed in Ka and Tmax and with a reduced variability) with a system that is known to be able to induce satiety is surprising and counterintuitive (as one would expect a much delayed Tmax due to the satiety). A longer Tmax would for most applications also be undesired. As mentioned, in the present invention, it is preferred that in the composition, method and use as specified above the drug or mixtures of drugs is a pharmacologically active compound, and even more preferably that it is present in an effective amount. Even more preferably, the concentration of the drug or mixtures of drugs concerned is such that the amount (or concentration) of the drug or mixtures of drugs in the foamed composition is such that 50 ml of said foamed composition comprises an amount of the drug or mixtures of drugs concerned equal to 10 to 300% of the effective amount for said drug or mixtures of drugs. This is preferred as it is believed that at and above 50 ml the effect of reducing variability is achieved (with stronger effects at 100 ml and above).

In the present invention, for predictable and reliable pharmacokinetics it is preferred that in the foamed compositions the drug or mixture of drugs is dissolved or dispersed in the continuous aqueous liquid phase of the foamed composition. The benefits of the present invention may be obtained with any type of edible foamed composition as specified for the composition according to this invention and for the method and uses of this invention, as long as it has sufficient foam stability in the mouth. It is believed in the context of this invention (i.e. for the desired effect on variability) that if a foam has sufficient stability in the mouth, such foam will also have sufficient stability in the esophagus and stomach to yield the advantages herein described. "Sufficient foam stability in the mouth" is herein defined as a reduction in overrun of less than 35% when a sample of the product is subjected to a stability test in which conditions of shear are applied that are similar to those observed in the mouth. The in-mouth stability of a foam composition of the present invention and in its methods and uses can be determined by introducing a predetermined volume of an edible aerated product in a glass funnel

(diameter 100 mm, neck length 100 mm, neck diameter 10 mm), which is connected to a silicone tube (length 400 mm, diameter 12x8 mm). The middle part of the silicone tube is inserted into a peristaltic pump (e.g. a Verderflex 2010, Verder Ltd, Leeds, UK) operating at 60 rpm. After the processing in the peristaltic pump the sample is collected in a glass measuring cylinder and the product volume and product weight are measured

immediately. In the shear test described above the foam products in the methods and uses of the present invention typically show a reduction in overrun of less than 30%, preferably of less than 25%, most preferably of less than 22%. In contrast, known edible foam products, such as whipped cream, show decreases in overrun that are well in excess of these percentages. According to a preferred embodiment, the product obtained from the in-mouth stability test described above still exhibits an overrun of at least 100%, more preferably of at least 120%, and even more preferably at least 150%. Edible foam products that are capable of retaining a high overrun when subjected to conditions of shear that are similar to those observed during mastication and preferably that further exhibit high stability under gastric conditions are extremely useful for the purposes of this invention. According to a particularly preferred embodiment, the aforementioned criteria are also met by the foam products in the present composition, method and uses of the invention if the shear stability test is conducted at a temperature of 37°C, thus reflecting the prolonged in-mouth stability of the product under conditions of shear that are similar to those exerted during mastication.

More preferably, the foamed composition of the present invention, including in its uses and methods, has a physical (foam) stability such that the foam has a half life in the stomach of at least 20 minutes, preferably at least 30 minutes, more preferably of at least 45 minutes. "Foam half life in the stomach" herein is the gastric retention time where 50% of the foam volume ingested remains present as a foam in the stomach. The presence of a foam in the stomach, and thus the half life, can be determined by visualisation techniques as known in the medical profession. Of these, Magnetic Resonance Imaging (MRI) or Computer Tomography (CT) scanning are preferred techniques, as they directly show the presence of foam, air and liquid. Ultrasound imaging can also be used for this, but due to differences in image quality and the interpretation of it a large enough set of test persons would be needed, as a person skilled in the art of ultrasound imaging would know. Also, with ultrasound imaging a foamed composition in the stomach as such cannot be visualised using ultrasound imaging, but the presence of foam can be derived from the reappearance of antral motility and ultrasound signal after the foam has left the stomach. Also, these imaging techniques can also be used to determine whether a foamed composition has a sufficient stability to pass the mouth and be present for some time as an aerated composition.

Even more preferably, the foamed compositions of the present invention, including in its uses and methods, have a very high gastric stability. Such high gastric stability of the foamed product can be apparent from the time (t _½ ) needed to achieve a reduction in overrun of 50% under simulated gastric conditions. The foamed product of the present invention exhibits a t _½ of more than 30 minutes. The aforementioned parameter t _½ is determined in a gastric stability test involving transferring 400 ml of an aerated edible product to a vessel of a United States Pharmacopeia (USP) dissolution model II apparatus (VanKel VK 7000), of which the temperature of the water bath is set to 37.5 °C and slow shear in the stomach is simulated with a special paddle at a stirring rate of 1.2 s-1 (72 rpm). Twenty five ml of fasted state gastric fluid, containing 80 μΜ sodium taurocholate, 20 μΜ lecithin, 0.1 mg/ mL pepsin, 0.1 mg/ mL amano lipase A, 34.2 mM sodium chloride in water and set to pH 1 .6 using hydrochloric acid, is subsequently carefully added along the vessel wall. A Masterflex® US pump is started simultaneously to pump simulated gastric fluid into the vessel at a rate of 0.8 ml/min. With the same rate, fluid from the bottom of the vessel is also removed. The foamed composition and the liquid volumes are read at start, after 5 min. and 10 min., and further at 10 min. intervals up to 60 minutes. The benefits of the foamed product in the present invention are particularly pronounced in case the gastric stability is very high. Accordingly, in a particularly preferred embodiment t _½ exceeds 45 minutes, even more preferably it exceeds 60 minutes, even more preferably it exceeds 90 minutes and most preferably t _½ exceeds 120 minutes.

The compositions of the present invention and in the method and uses herein are such that they have an overrun of at least 100%. According to a preferred embodiment, the edible foamed product of the present invention has an overrun of at least 120%, more preferably of at least 150%, and even more preferably between 150% and 800%.

The gas phase in the present product can comprise air or any other gas that is considered safe for food applications.

The foam in the compositions, method and uses of the present invention is a pourable or spoonable aerated composition. According to one embodiment, the product is non- pourable (i.e. spoonable). Such a spoonable product typically exhibits spoonable rheology defined as follows: yield value of >50 Pa, when extrapolating from shear rates between 100 and 300 s ^"1 , a Bingham viscosity <500 mPa.s between shear rates of between 100 and 300 s ^"1 , a failure at stress at a strain of <0.5 Radians. The yield stress is determined at a temperature of 20 °C using a Haake VT550 viscometer. According to another embodiment, the edible foam product is pourable. A pourable product offers the advantage that it can be drunk. If the product is drunk rather than eaten, the chance of undesirable density increase as a result of mastication is minimised - for example bread is high overrun product, but practically all air is lost during mastication. The foam in the compositions, methods and uses of the present invention is not a mousse and is not sponge-like. A mousse is herein to be understood as having a continuous phase that is a gelled phase, and this is to be distinguished from the currently used pourable and spoonable aerated compositions, in which the continuous phase of the aerated composition is not a gelled phase (but a liquid phase). The foam in the compositions, methods and uses of the present invention is preferably non-frozen when ingested, as frozen compositions like ice cream tend to melt in the mouth and/or upper gastro-intestinal tract and following that lose their foam stability.

The foam in the compositions, methods and uses of the present invention preferably comprises by weight 50-99.5% water, a foaming agent and a stabiliser (next to the pharmacologically active component).

In this connection, the foaming agent preferably comprises, for a good foamed

composition, one or more of:

- a food grade water-soluble emulsifier having an hydrophilic-lipophilic balance

(HLB) value of at least 8, preferably at least 9, more preferably at least 12, a food grade protein;

food grade amphiphatic particles having a contact angle at air/water interface between 70 and 120 degrees, and preferably having a volume weighted mean diameter of 0.02 to 10 micron (μηι).

Examples of preferred food grade water-soluble emulsifier having an HLB value of at least 8, preferably at least 9, more preferably at least 12 herein are: sodium docecyl sulfate (SDS), SSL, polyoxyethylene 20 sorbitan monolaurate = polysorbate 20 (Tween 20), polysorbate 40 (Tween 40), polysorbate 60 (Tween 60), Molec MT (enzymatically hydrolysed lecithin) and L1695 (lauric ester of sucrose, ex Mitsubishi-Kasei Food Corp.), and DATEM (diacetyl tartaric acid ester of monoglyceride).

Preferred food grade proteins in this connection comprise dairy proteins such as whey protein and/or casein protein and sources thereof, as well as vegetable proteins like soy protein, meat- and fish derived protein, and egg protein like albumin. When used as sole foaming agent, such food grade proteins are preferably used in an amount of from 1 to 7% by weight. Preferred food grade amphiphatic particles herein comprise one or more of cocoa particles. As to the stabiliser, e.g. to give the product sufficient physical stability, e.g. to allow some time between preparation of the foamed composition, it is preferred that the stabilizer comprises a dietary fibre or a sucrose ester. Preferred amounts in this context are: from 0.1 to 5% by weight. Too little may not provide the desired stability, too much may make foaming difficult. Suitable dietary fibres in this context are one or more of the group consisting of: carrageenan, xanthan, cellulose, gellan, locust bean gum, with xanthan being the most preferred stabiliser (as it provides stabilising without too much viscosity increase).

Fat may be present in the compositions, methods and uses according to this invention, but such is preferably kept at a low level, so as not to induce too much calories to the composition. Also, fat may act detrimental on the stability of the aerated compositions. Hence, in the compositions in the methods and uses herein, the edible foamed

composition comprises fat in an amount of less than 2% by weight, preferably less than 1.8% by weight, more preferably between 0 and 1.8% by weight, even more preferably between 0 and 1 .5% by weight, even more preferably form 0.01 to 1.5% by weight.

Next to the foaming agent, stabiliser, water and optionally fat, other components that may be present include carbohydrates, (non-caloric) sweeteners, flavouring components.

The edible foamed compositions of the present invention, and in the methods and uses of this invention as specified herein, can be prepared by any suitable means. The foamed compositions may be manufactured, packed and marketed in a foamed form, but it is also possible to prepare a non-aerated product which is packed and marketed, which is then foamed some time or immediately before consumption, either by the individual or at a point of sale or distribution. A convenient way (and one which can easily give foamed compositions of high stability) to offer such to users is when the composition according to this invention and for use in the method and uses of this invention is packed as a non- aerated (e.g. liquid) composition in a pressurised container in a liquid form. By this, the pressurised container can hold the edible liquid (non-aerated) composition and a propellant, which liquid composition can be released from the container by activating a valve (on the container) to produce an edible foamed product. Hence, more preferably, the invention further relates to the use in the composition, method and uses of the present invention of a pressurised container further comprising a propellant, and wherein the pressurised container is equipped with a valve, wherein the liquid can be released from the pressurised container by activating said valve to produce the aerated composition for the compositions, methods and uses according to this invention. Typically, the edible aerated product thus obtained has a density that is much lower (e.g. 40% lower) than that of the liquid composition in the container. According to a preferred embodiment, the edible foamed product produced upon activation of the valve has the same composition as the edible liquid composition (gas phase not being included). Suitable propellants in this include compressed gases, especially liquefied gasses.

Preferably, the propellant employed is selected from N ₂ 0, N ₂ , C0 ₂ , air and combinations thereof. Most preferably, the propellant employed is selected from N ₂ 0, N _2i C0 ₂ and combinations thereof. Typically, the propellant contained in the pressurised container has a pressure of at least 2 bar, more preferably at least 3 bar. Usually, said pressure does not exceed 12 bar.

Whether the foamed composition in the compositions, methods and uses as set out herein is packaged in a pressurised container or not, it is preferred in the present invention that the foamed composition is prepared by the user from a composition which is packaged as a liquid. This liquid is then to be turned into the aerated composition before being consumed.

Alternatively, the foam of the present invention may also be prepared by whipping a suitable liquid using, for instance, a standard kitchen mixer. By this, an amount of the liquid is added to the bowl of the mixer and mixed at high speed to aerate the

product. Typically, the edible aerated product thus obtained has a density that is much lower (e.g. 40% lower) than that of the liquid composition. Through varying the amount of liquid added to the bowl of the kitchen mixer and by varying the mixing time and speed, different levels of overrun can be achieved.

The gas bubbles contained within the edible foamed composition in the compositions, methods and uses according to this invention can vary widely in size. Typically, the air bubbles in the product have a volume weighted mean diameter in the range of 5-500 μηη, preferably of 10-200 μηη. The volume weighted mean diameter of the gas bubbles is suitably determined by means of optical microscopy.

The stability of the edible foamed product, especially if it is produced in situ from a pressurised aerosol system, is affected by the composition of the gas that is retained within the foamed product. In order to generate a very stable foamed product, it is advantageous to include a gas that has limited water-solubility. Air, for instance, is not particularly suitable as e.g. oxygen has a relatively high solubility in water. According to a particularly preferred embodiment, the edible foamed product in the present invention contains a gas that is less soluble in water than air (at a temperature of 37°C. According to another preferred embodiment, relative to air, the gas contained in foam product contains elevated levels of one or more of the following gasses: N ₂ , N ₂ 0, C0 ₂ , He, 0 ₂ . Here the term "elevated" means that the concentration of at least one of said gasses is at least 10% higher than in air.

The foam can be designed to contain calories or not, depending on the drug, the desired effect and the use. When only a satiety effect is desired for the drug(s) concerned, then the foam is preferably designed as a low calory foam.

Hence, it is preferred that the composition for use in the current method is has a low caloric density. Caloric density of the aerated composition used in the current methods herein is digestible (for humans) calories per volume (or weight, depending on what is specified) of aerated compositions. Hence, in the current methods, it is preferred that in the methods and uses of the present invention, the aerated composition has a caloric density of less than 5 kcal/ml, preferably less than 2 kcal/ml, more preferably less than 0.5 kcal/ml of aerated composition (ready for consumption). In this connection, it is preferred for this invention that sucrose and glucose do not count as drugs. In the prior art it is known that uptake of drugs can be sustained or controlled by the addition of a material to the drug that causes a delayed or sustained release of such drug. For example, a drug may be coated with a material which degrades slowly in the intestinal tract, to sustain release of the drug. Alternatively, the drug may be coated with a material which is insoluble under the acidic conditions of the stomach, in order to control uptake of the drug in the intestines. Or vice versa, the drug may be coated with a material which is soluble in the stomach, in order to facilitate the uptake of the drug in the stomach. One of the advantages of the current invention, is that addition of materials which normally delay or control uptake of drugs are not required to be used in the current compositions of the invention. Therefore preferably the foamed compositions of the present invention are free from a biopolymer or a bioengineered composition or a polymer for providing a sustained or delayed release of a medicinal or nutritive component. EXAMPLES Example 1

Exemplary aerated edible product formulations with BSC class I drugs.

Table 1. Compositions of formulations for producing aerated edible products with BSC class I drugs.

A B C D E

Ingredient % (w/w) % (w/w) % (w/w) % (w/w) % (w/w)

Skimmed milk powder 13.6 13.6 13.6 13.6 13.6

Xanthan gum 0.5 0.5 0.5 0.5 0.5

Orange syrup 10 10 10 0 0

Water 75.9 75.9 75.9 85.9 85.9

Caffeine 0 0 0 0 0.007

Paracetamol 0.035 0.017 0.007 0.017 0

The liquid formulations were prepared as follows. All ingredients are added to water and mixed until fully dispersed.

Example 2

Exemplary aerated edible product with BSC class I drug.

Three hundred gram of formulation B (example 1 ) was poured into a pressurisable dispenser (cream whipper "Gourmet Whip", 0.5 I, iSi GmbH). The dispenser was closed and a N ₂ 0 gas charger was mounted on the dispenser until gas was released therefrom. The dispenser was vigorously shaken for about 20 seconds, following which the contents are released yielding an aerated edible product.

This example shows that foams containing drugs can be produced by a pressurised can. Example 3

Exemplary aerated edible product with BSC class I drug.

Three hundred gram of formulation B (example 1 ) gram was weighed in the metal bowl of a kitchen machine (Kenwood, model chef classic KM330). The liquid was mixed at maximum speed for 1 min after which an overrun of 250% is achieved.

This example shows that foams containing drugs can be obtained without pressurised cans. Example 4

Release of BSC class I drug from aerated edible products.

Aerated edible product containing paracetamol (=acetaminophen) (example 3) was put under mild shear in a vessel of a United States Pharmacopeia (USP) dissolution model II apparatus (VanKel VK 7000) to disrupt the aerated edible product. Samples of the aerated edible product and drained liquid were collected and analysed for paracetamol content. For the paracetamol analysis, 5 ml of each sample was weighed into a 50-ml volumetric flask and 25 ml acetonitrile was added for precipitation of the proteins. The samples were vortexed for 30 sec and then placed in an ultrasonic bath for 10 min. After cooling down the flasks were filled to the mark with acetonitrile, homogenized, and centrifuged at 3000 rcf for 10 min. Supernatants were diluted prior to HPLC analysis. All samples were analyzed in duplicate.

Samples were analyzed using reversed-phase HPLC (Shimadzu LC 10 ADvp) equipped with a Luna Phenyl hexyl (4.6 x 250 mm, 5 μηη) (Phenomenex) column. Flow rate was 1 ml/min and injection volume was 10 μΙ. The column temperature was kept at 35°C and the UV detector (Shimadzu UV) was set at 250 nm wavelength. Solvent A consisted of 2% acetic acid in water containing 20 mg/l EDTA. Solvent B consisted of 2% acetic acid in acetonitrile. Table 2 shows the gradient that was applied for the analysis. Quantification of the paracetamol contents in the samples was performed using an external calibration curve of paracetamol.

Table 2. Gradient for the HPLC analysis of paracetamol in aerated edible product samples and liquid samples

Time (min) Solvent B (%)

0 5

10 5

15 7

15.1 98

25 98

25.1 5

45 5 The results are shown in table 3. The results confirmed a stabile distribution of paracetamol over time in the aerated edible product and the drained liquid (ratio very close to 1.0). Table 3. Paracetamol analysis (mg/g, via HPLC) in drained liquid and remaining aerated edible product, taken from formulated aerated edible product put under mild shear over 120 min. Added amount of paracetamol was 1.74 mg/g.

paracetamol (mg/g) paracetamol

Time (min) Aerated edible Liquid ratio aerated edible product product/ liquid start 1.74 1 .49 1 .17

30 1.68 1 .73 0.97

60 1.66 1 .76 0.94

120 1.74 1 .70 1 .02

Mean ± SD 1.71 ± 0.04 1.67 ± 0.12 1.03 ± 0.10 This example shows that there is no preferential drainage of the paracetamol into the drained liquid: the system remains homogeneous with respect to the paracetamol concentration. The small amount of drained liquid formed after whipping and before dosing (extremely low volume) still contained the same concentration of paracetamol. Example 5

Ambient stability of aerated edible products containing BSC class I drug.

Eight hundred ml of aerated edible product from example 3 was transferred into a measuring glass cylinder which had been previously tared on a balance. Total aerated edible product mass was measured. The top of the cylinder was subsequently covered with parafilm to prevent evaporation. The measuring glass cylinder was placed on a lab bench at ambient temperature and total aerated edible product volume and drained liquid was monitored for a period of at least 60 minutes. No significant changes in terms of aerated edible product collapse, severe creaming, severe disproportionation or

combinations thereof occurred.

This example shows a paracetamol-containing foam having a good bench-top stability.

Example 6

In mouth stability of aerated edible products containing BSC class I drug

Four hundred ml of aerated edible product from example 3 was put into a glass funnel (diameter 100 mm, neck length 100 mm and a neck diameter of 10 mm). The funnel was connected to a silicone tube with a length of 400 mm and a diameter of 12x8 mm. The middle part of the silicone tube was inserted into a peristaltic pump (Watson-Marlow model 501 ) and operating at 60 rpm. Aerated edible product pumped through the silicone tube was collected in a previously weighed vessel. The impact of the simulated oral cavity test is determined by observing the start (400 mL) and end volume. The simulated oral cavity test reduced the overrun of the aerated edible product by less than 5%.

The same experiment was performed with an aerated edible product based on formulation C in example 1 and aerated as described in example 3. Also for this aerated product, the simulated oral cavity test reduced the overrun of the aerated edible product by less than 5%.

This example shows a paracetamol-containing foam of sufficient oral stability. Example 7

In vitro gastric stability of aerated edible products containing BSC class I drug.

After collecting aerated edible product from the mouth stability test (example 6), 400 ml was transferred to a vessel of the USP dissolution model II apparatus. The temperature of the water bath was set to 37.5 °C. The slow shear in the stomach was simulated with a special paddle at a stirring rate of 1 .2 s ^"1 (72 rpm). The vessel was weighed and placed in the USP model and the stirring was started. Twenty five ml of fasted state gastric fluid (Table 4) was pipetted carefully along the vessel wall. A Masterflex® US pump was started simultaneously to pump simulated gastric fluid into the vessel at a rate of

0.8 ml/minute. With the same rate fluid from the bottom of the vessel was also removed. The aerated edible product and the liquid volumes were read at start (t = 0, after 5 min. and 10 min., and further at 10 min. intervals up to 60 minutes.

Table 4. Composition of fasted state gastric fluid, pH=1.6

Ingredient Amount per 500 ml

Sodium taurocholate (80 μΜ) 21 .52 mg

Lecithin (20 μΜ) 7.6 mg

Pepsin ( 0.1 mg / mL ) 50 mg

Amano lipase A 50 mg

Sodium chloride ( 34.2 mM) 1 gram

Hydrochloric acid to set pH to 1.6

Deionized water Fill up to 500 ml Figure 1 shows the stability of the aerated edible product from example 5 under simulated gastric conditions. Less than 40% of aerated edible product is digested within 60 min under the conditions used in this test. The t _½ is 55 minutes.

This example shows a paracetamol-containing foam of good in-vivo gastric stability. Example 8

In vivo gastric stability of the aerated edible products.

Volunteers and study design

The study was a single-centre, randomized, placebo-controlled, balanced cross-over design consisting of a screening visit and three test days. The volunteers were recruited from local area of Nottingham. Selection criteria were: age 18-60 years, Body Mass Index (BMI, kg/m2) >20 and <35, apparently healthy (measured by questionnaire) and not using medicines judged likely to influence the study results. Only normal and low restraint eaters (Federoff et al, 2003; Polivy et al. 1978) were included. Any subjects with tendencies toward diagnosable eating disorders (anorexia nervosa or bulimia) were excluded

(Morgan et al., 1999). In total 20 subjects were screened to take part in the study of which 1 was not eligible and one withdrew after successful screening and before his first study day. From the eligible participants identified, 18 were randomised to take part in the study. Following the blind review of the study data, none of the subjects were withdrawn from the analysis, however, one subject only attended one test day due to a change in his diary commitments and data of another subject for one test day were not used for analysis as this subject was not fasted at that day. Characteristics of the 18 subjects were: age: 24.9 (range 19 - 38) y; BMI: 23.8 (range 20.3 - 29.5) kg/m2.

Volunteers were scanned on arrival at the centre to ensure their stomachs were empty (save resting gastric juices). Each day the subjects received one out of two isocaloric test products. An aerated beverage that was expected to be stable in the stomach ("Stable Aerated edible product") and an aerated beverage that was expected to be less stable in the stomach ("Less Stable Aerated edible product") (for details see below and example 9).

Subjects were scanned at baseline to ensure their stomach was empty. They then consumed 150g of one of the products instead of breakfast and MRI measurements were conducted at several time points (up to 4 hours after consumption of the test product) to measure gastric behaviour. The subjects were blind to aerated test product type.

The participants were instructed to minimise changes in their physical activities and were not allowed to follow a diet one month prior to and during the test period. On the day before the test day the subjects were instructed not to use alcohol or play sports. They were asked to refrain from consuming any food or drink other than non-caloric beverages from 22.00 until arrival at the test facility. During the test day, the volunteers were instructed to avoid high intensity physical activity and direct contact with food. Volunteers were not allowed to eat and drink anything else during the study. Subjects were asked about their mode of transportation together with food consumption the evening before the test day and drinks consumed from 10 pm till the next morning.

Test products

Two test products were evaluated:

1. 490 ml aerated drink comprising of 140 ml protein/carbohydrate beverage (13.6

wt% SMP, 0.5 wt% xanthan gum, 10 wt% lemon syrup, water) with an additional 350 ml of air (= total volume 490 ml, "Stable Aerated edible product")

2. 490 ml aerated drink comprising of 140 ml protein/carbohydrate beverage (13.6

wt% SMP, 0.1 wt% xanthan gum, 10 wt% lemon syrup, water) with an additional 350 ml of air (= total volume 490 ml, "Less Stable Aerated edible product")

The test products were equicaloric (1 10kcal) and their nutritional composition is detailed in Table 5.

Table 5. Nutritional composition of test products

volume weight Energy ³ Fat Protein Carbohydrate

of which of which

Total ^b sugars fibres

[ml] [g] [kcal] [g] [g] [g] [g] [g]

Stable Aerated 490 150 1 10 0.2 7.2 20.2 18.6 0.8 ^c edible product

Less Stable 490 150 108 0.2 7.2 19.6 18.6 0.2 ^d

Aerated edible

product

Key: ^a based on energy content of fibres of 4 kcal/g

b total carbohydrates, including sugars and fibres

c contains 0.8 g xanthan gum

d contains 0.2 g xanthan gum

Both aerated edible products were produced by aerating the beverage with air as described in example 3. All test products' overrun was measured and amounted 247% ± 7% for the stable and 254% ± 3% less stable aerated edible products. For both test products, the simulated oral cavity test as described in example 6 reduced the overrun of the aerated edible products by less than 5%. Magnetic resonance imaging (MRI)

All MRI was carried out on a 1 .5 T Philips Achieva MRI research-dedicated scanner (Philips Healthcare) sited at the University Campus, Nottingham using a parallel imaging receiver coil wrapped around the abdomen. The BTFE, single shot MRI sequence used acquired 30 contiguous slices across the abdomen under a 12 seconds expiration breath- hold. The final, optimised sequence parameters were: 80° flip angle, TR=2.8 ms, TE=1 .40 ms, acquired resolution of 2.00 mm x 1.77 mm x 10 mm (reconstructed to 1.56 mm x 1.57 mm x 10 mm). This was run in the transverse (axial) plane. For the coronal ('frontal view') single shot, fast spin echo sequence (as used in MRCP) the sequence parameters were: multi-slice 2D single shot imaging, TEeff = 320 ms, TReff =∞, 24 slices, SL = 7 mm, FOV = 400 mm, RFOV = 90%, scan percentage = 80%, acquired voxel size=1 .56 x 2.83 x 7mm3 (interpolated to 0.78 ^χ 0.78 * 7mm3), total scan time = 24 s acquired in a single breath-hold.

The subjects were supine on the scanner bed. They were kept tilted with their left side slightly raised on the scanner bed using a folded towel to help avoiding the possibility that floating layers of the test meals could empty first through the pylorus, which would be different from what happens during normal upright digestion.

The validity of MRI measurements of gastric volumes (from single scans) and of gastric emptying has been validated against intra gastric balloon infusion (Boulby et al., 1997) and against double marker indicator (Schwizer et al., 1994) and gamma scintigraphy (Feinle et al., 1999).

Gastric aerated edible product volume was measured from the balanced gradient echo (also called balanced turbo field echo or BTFE). At each time point 30 axial, 2D slices of the abdomen were acquired. Every data set from each time point was then recalled on a UNIX workstation for analysis. On such axial sections, commercial specialistic software (Analyze 9, Biomedical Imaging Resources, Mayo Clinic, Rochester, MN) was used to trace manually on each 2D slice around the region of interest (ROI) of the aerated edible product in the stomach on each slice. These series of 2D ROIs were then collected together as a 3D total volume and saved as a text file for input into an Excel database. Statistical analysis

Analyses were carried out using both ITT (Intention To Treat) and PP (Per Protocol) populations. Analysis for ITT and PP had similar results and conclusions. Therefore only the ITT results are reported here.

Results

Gastric aerated edible product volume

There was good contrast in the MRI images for all 2 products, with the three regions of air, aerated edible product and liquid clearly defined (Figure 2). On the T2 weighted BTFE images acquired in this study, liquids appear bright white. The aerated edible products appeared naturally darker than fluid and than other organs like the spleen, due to the aerated edible product's lower water proton density and air bubble matrix affecting the signal itself. As such, the images' intensity display scale was increased for better visualisation of the aerated edible products, for display, and for analysis.

All three phases could clearly be distinguished on the images and measured and the regions of interest (ROIs) were converted to volumes. Figure 2 shows examples of ROIs drawn intragastrically for a Stable Aerated edible product and a Less Stable Aerated edible product at different time points. These were plotted in Microsoft Excel to allow for observations on the time courses of intragastric volumes.

The intragastric aerated edible product volumes mean time series are shown in table 6. At t=10 min after intake the volume of aerated edible product for the Stable Aerated edible product test product was significantly higher than for the Less Stable Aerated edible product test product. After that postprandial time point and at all time points up to T=90 min the mean intragastric aerated edible product volumes for the Stable Aerated edible product were also higher than those for the Less Stable Aerated edible product. The most significant differences were found within the first 70 min after treatment intake.

Conclusions

The study showed that MRI is very appropriate to measure serially separate volumes of aerated edible product after ingestion of aerated products. The decrease in aerated edible product volume was clearly slower for the Stable Aerated edible product as compared to the Less Stable Aerated edible product. Table 6 Stomach aerated edible product content (LSmeans, in ml) in time for the different treatments (less stable aerated edible product, stable aerated edible product).

Treatment

Less Stable Aerated edible product Stable Aerated edible product

Time Volume Standard Volume Standard

(ml) Error Lower Upper (ml) Error Lower Upper

-15 0.0 0.0

10 373.3 14.2 344.6 402.1 444.4 13.7 416.6 472.2

30 102.3 16.4 69.2 135.3 309.8 15.8 277.8 341 .7

50 25.8 13.1 -0.6 52.2 160.9 12.6 135.4 186.4

70 9.5 9.8 -10.4 29.3 63.1 9.5 43.9 82.3

90 3.5 3.2 -3.1 10.0 12.1 3.1 5.8 18.4

120 0.3 0.5 -0.7 1 .3 1.2 0.5 0.2 2.1

150 0.1 0.1 -0.2 0.4 0.3 0.1 -0.0 0.5

180 -0.0 0.1 -0.1 0.1 0.1 0.1 -0.0 0.2

Example 9

5 Correlation in vitro and in vivo gastric stability of the aerated edible product.

Study products used in the MRI study (example 8) were tested for in vitro gastric stability as described in example 7. The overrun of the aerated edible products was 250%. The run time of the gastric step was 50 minutes. The results are given in figure 3. Figure 4 shows the linear regression between the data points from the MRI study and the in-vitro0 gastric stability test. The results clearly show that the in vitro model for gastric stability correlates well with the in vivo gastric stability of aerated edible products.

Example 10

Dose dependency of aerated edible products on satiety.

5

Study design

The study used a random allocation, parallel design, with treatments balanced across test days. Each subject group was given a single exposure to a single volume of an aerated edible product, each portion having a volume of 10, 25, 50, 100, 150 or 250 ml. This0 product was given as a mid-morning snack (at 10.30 am) following a fixed 250 kcal breakfast given at 08.00 am. Self-reported eating motivation ratings (6 scales) were collected regularly from 155 minutes prior to consumption of the test product and for 3 hours afterwards.

Subjects

Healthy normal weight and overweight male and female participants (age 18-50 yr, BMI 20-32 kg/m ² ) were recruited from local area of the research centre. Only normal and low- restraint eaters were included, based on the Revised Restraint Scale (Polivy et al., 1978; Federoff et al., 2003). Any subject with a tendency toward a diagnosable eating disorder (anorexia nervosa or bulimia) was also excluded based on the SCOFF questionnaire (Morgan et al., 1999). From the eligible participants identified, 144 were admitted onto the study. Potential volunteers were trained on completion of visual analogue scales (VAS) for subjective ratings of ingestive behaviour, and were familiarized with the test product and the study design.

The 144 participating subjects were randomized into groups of 24 subjects per treatment, with groups matched for gender mix, age and body weight (mean within 5 yr and 5 kg). Ten subjects were withdrawn from the study for reasons unrelated to the study products. Characteristics of the remaining 133 subjects (91 females, 42 males) were: age: 35.8 (range 18 - 60) y; BMI: 24.8 (range 21.0 - 34.6) kg/m ² . Study products

Each subject was given a single exposure to a single portion of an aerated edible product at a specified volume at 10.30 following a fixed breakfast at 08.00. Six aerated edible products were evaluated varying in total volume.

The test products consisted of Slim-Fast Optima high protein ready-to-drink meal replacement shakes (190 kcal/325 ml when not aerated), aerated on site with N ₂ 0 (from an iSi dispenser and using an iSi N ₂ 0 disposable gas filled cylinder) (Slim-Fast is a trademark of Unilever PLC, United Kingdom and Unilever NV, Netherlands; iSi is a tradename of iSi GmbH). An ingredients list of the non-aerated SlinvFast high protein chocolate RTD shake base is shown in Table 7. The liquid formulation used was the same as the commercial product identified above, but with a different chocolate flavouring component. The overrun of the product was approximately 200%. This means that the energy content per serving was approximately 2, 5, 10, 19, 29 and 48 kcal for the 10, 25, 50, 100, 150 and 250 ml aerated servings (corresponding to approx. weights of 4, 1 1 , 18, 35, 50 and 85 gram servings), respectively. Table 7. Nutrient composition of Slim -Fast High Protein Extra Creamy Chocolate RTD shake (US formulation)

Amounts per can (325 ml)

Calories 190

Total Fat 5g

Saturated Fat 2g

Cholesterol 10mg

Sodium 220mg

Potassium 600mg

Total Carbohydrate 24g

Dietary Fiber 5g

Sugars 13g

Protein 15g

+ Vitamin-Mineral complex

Ingredients: Fat Free Milk, Water, Calcium Caseinate, Milk Protein Concentrate,

Maltodextrin, Cocoa (Processed with Alkali), Canola Oil, Gum Arabic, Cellulose Gel, Sugar, Mono and Diglycerides, Fructose, Potassium Phosphate, Soybean Lecithin, Cellulose Gum, Carrageenan, Artificial Flavor, Isolated Soy Protein, Sucralose and Acesulfame Potassium (Non Nutritive Sweeteners), Dextrose, Potassium Carrageenan, Citric Acid and Sodium Citrate. Vitamins and Minerals: Magnesium Phosphate, Calcium Phosphate, Sodium Ascorbate, Vitamin E Acetate, Zinc Gluconate, Ferric

Orthophosphate, Niacinamide, Calcium Pantothenate, Manganese Sulfate, Vitamin A Palmitate, Pyridoxine Hydrochloride, Riboflavin, Thiamin Mononitrate, Folic Acid, Chromium Chloride, Biotin, Sodium Molybdate, Potassium Iodide, Phylloquinone (Vitamin K1 ), Sodium Selenite, Cyanocobalamin (Vitamin B12) and Cholecalciferol (Vitamin D3).

At least 24 hours before the test day the SlinvFast high protein chocolate RTD was stored at 5 °C, while the iSi N ₂ 0 gas filled cylinder and dispensers were stored at room temperature. All test products were presented in an accompanying beaker. All aerated edible products were consumed with a 10 ml black plastic spoon and the subjects were instructed to eat all of the aerated edible product within 10 minutes. All test products were prepared on the test days, according to a standard operating procedure. In short, the content of one 325ml can of Slim-Fast high protein chocolate shake was poured into the stainless steel iSi bottle and the device head was screwed onto the stainless steel bottle One iSi N ₂ 0 gas filled cylinder was inserted into the cylinder holder and the cylinder holder was screwed to the device head until all of the content of the cylinder was released into the bottle. Thereafter the device was vigorously shaken for 20 seconds. The foam was then dispensed by turning the device upside down with the decorator tip in the vertical position and gently pressing the lever. The entire amount (~900ml) was dispensed 5 against the inside edge of a large glass container and then the required foam volumes was poured into glasses which had been pre-marked with the required volume. The weight of the foam was subsequently measured.

Subjective feelings of hunger and satiety

10 Self-report ratings for appetite measures were collected at time points of -155, -120, -90, - 60, -30, -5, 15, 30, 60, 90, 120, 150, and 180 min (where test product consumption at 1030 was regarded as 0 min). Ratings of satiety feelings were scored using reproducible and valid scales (Stubbs et al., 2000; Flint et al. 2000) by means of a mark on 60-mm scales using EVAS (Electronic Visual Analogue Scale, iPAQ; Stratton et al. 1998) (iPAQ is

15 a trademark of Hewlett Packard, USA) anchored at the low end with the most negative or lowest intensity feelings (e.g., not at all), and with opposing terms at the high end (e.g., very high). Volunteers were asked to indicate on a line which place on the scale best reflects their feelings at that moment. The scale items were "desire to eat a meal", "desire to eat a snack", "hunger", "how much do you want to eat", "satiety" and "fullness".

20

Analyses

The study was a product benchmarking study, aimed to generate a dose-response profile for satiety effects, focused on identifying lower volume limits for potential consumer concepts. Curves of Least Square means (LSmeans) were produced based on the 25 measurements, and based on these curves a time-to-return-to-baseline (TTRTB) was calculated by using a modeling technique based on the Weibull distribution (Schuring et al 2008). This Weibull method also turned out to be the most suitable, non-parametric model to estimate TTRTB for these satiety curves.

30 Results

The consumption of small portions of aerated edible product in between meals (i.e. as a snack) induced clear effects on eating motivational ratings. The results indicate a rough dose-response, although this is not completely consistent across the different line scales used. Strongest, robust effects are shown for 250 ml, while no effect is observed for the 35 10 or 25 ml (see Tables 8, 9 and 10). For 10 and 25 ml the TTRTB estimates could not be calculated, as the curves did not cross the baseline. This is consistent for all line scales. Intermediate effects of aerated edible product on feelings of hunger and satiety were observed for the 50, 100 and 150 ml. For all line scales the 50, 100 and 150 ml volumes showed greater responses as compared to the 10 and 25 ml. Table 8. Time to return to baseline (TTRTB) for 'Desire to eat a meal'

¹ TTRTB (minutes) by Weibull modeling

Table 9. Time to return to baseline (TTRTB) for 'Desire to eat a snack'

¹ TTRTB (minutes) by Weibull modeling

Table 10. Time to return to baseline (TTRTB) for 'Hunger'

¹ TTRTB (minutes) by Weibull modeling

Conclusion

A previous study has shown that aeration of liquid meal replacements leads to a high magnitude and duration of hunger suppression (increased satiety), which is substantially greater than non-aerated control products (De Groot et al., 2008; Blijdenstein et al., 2008) and also greater than examples in literature (e.g., Rolls et al., 2000; Osterholt et al., 2007). In that study the foam was consumed as a breakfast. It was, however, unknown, at what minimum volume a meaningful effect on satiety could still be observed. A quick estimate of response profiles to 6 foam volumes (10-250 ml) was therefore established. The results indicate a rough dose-response. Strongest, robust effects are shown for 250 ml, while no effect is observed for the 10 or 25 ml. This was consistent for all line scales. Surprisingly, effects were also observed for the 50, 100 and 150 ml where intermediate effects were seen. In 4 out of 6 line scales the effect of 150 ml on peak and duration is somewhat more pronounced than the 50 and 100 ml, but for all line scales the 50, 100 and 150 ml volumes showed greater (and longer) responses as compared to the 10 and 25 ml.

The 50 ml and 100 ml contained only 10 and 19 kcal per serving respectively, yet showed meaningful effects on appetite. The effects on appetite observed here are greater (and persist for longer) than shown in literature for beverages having either no caloric content or a caloric content which is higher than the foams now tested, at the same volume. Peters et al. (201 1 ) for instance tested a 100 ml minidrink as a snack and effects on hunger and appetite were comparable or even smaller compared to the effects seen here for the 50 or 100 ml foam, yet the 100 ml minidrink contained considerably more energy (80 kcal). Comparable or even smaller effects on appetite were also seen when testing 150 ml soup containing 150 kcal (Gray et al 2002) or 300 ml dairy-based drink containing 500 kcal (Rolls et al 2000). Although plain water or artificially sweetened water do decrease appetite, volumes needed are much higher and the temporal effect is much shorter as compared to the 50 or 100 ml foam (e.g. Monsivais et al., 2007)

The 250 ml volume led to an appetite response with an estimated TTRTB of 96 to 180 minutes, depending on the appetite rating used. Also the 100 and 150 ml volume produced a meaningful increase in TTRTB, generally between 45 and 93 min, depending on the appetite rating used. The 50 ml also generated a meaningful increase in TTRTB varying from 41 to 89 minutes, depending on the appetite rating used. For the 25 and 10 ml these values could not be estimated, as the curves did not cross the baseline (in the majority of subjects). TTRTB/kcal for hunger: 79/10, 80/19, 66/29 and 120/48 = 7.9, 4.2, 2.3 and 2.5 min/kcal for the 50, 100, 150 and 250 ml servings.

TTRTB/kcal for desire to eat a meal: 41/10, 45/19, 69/29 and 96/48 = 4.1 , 2.4, 2.4 and 2.0 min/kcal for the 50, 100, 150 and 250 ml servings.

Example 11 : study using foam containing paracetamol.

Objective

The aim of this study was to compare the PK profile of paracetamol when it was incorporated in an aerated drink (test, stable foam) versus a liquid (non-aerated) equivalent (control). GE half time (GE50) was the primary study parameter, and other parameters describing the PK profile of paracetamol were also determined. Study design

This study was an open-label, randomized, single-centre, explorative, cross-over design. The study consisted of a screening visit and two intervention periods. Treatments were separated by a wash-out period of 1 week. The study itself (i.e. after screening) covered 12 subjects.

On each test day the male volunteers (mean age: 30.4 year, BMI: 24.2 kg/m ² ) consumed, instead of breakfast, either an aerated or a non-aerated carbohydrate/protein drink at 08:00h. At 12:05h subjects received a standard lunch and at 14:05h and 16:05h subjects received a snack. Consumption of (non-caloric) drinks (with a maximum of 150 ml) was allowed two hours after test product consumption and every hour thereafter. The amount and type of food and drinks consumed during the first test day were recorded and repeated during the other test day.

At 18:30h subjects were allowed to leave the test facility. The subjects were divided in 2 groups, where one group followed the protocol, while the other group followed the same protocol, but one hour later. Subjects remained fasted from 22:00h the evening before test day until test product consumption at 8.00h (or 9.00h) at the test center. Venous blood was frequently sampled via an intravenous cannula and analyzed for paracetamol. In addition blood drops via finger prick were collected (DBS) at several time points in 6 subjects. These DBS was also analyzed for paracetamol. For an overview of timings of study measurements see table 1 1.

Legend:

(a) Test product will be administered at approximately 8:00 h.

(b) Period 1 only

(c) 5 minutes after blood sampling subjects get a finger prick for Dry Blood Spot sample (d) Hemoglobin

(e) non-caloric drink

Treatments

On test days subjects consumed either the foam (test) or the equivalent liquid (control):

• Control: 144 g non-aerated protein/carbohydrate beverage containing 250 mg

paracetamol (volume: 147 ml_).

• Test (foam): the same 144 g protein/carbohydrate beverage containing 250 mg

paracetamol with an additional -330 mL of air (= total volume -475 mL foam)

An in vitro gastric stability test showed that addition of the required dose of paracetamol (250 mg) did not affect the stability of the foam and as such can be used as a marker for GE. Table 12 describes the nutritional composition of the used liquid. During study days paracetamol was added to the liquid and foam. The foams were prepared in the same way as in example 3.

Table 12: Nutritional composition of liquid and foam

The composition of the formulations, either aerated (test) or non-aerated (control) were the following:

13.6% Skimmed Milk Powder

0.5% Xanthan Gum

75.9% water

10.0% orange syrup

Paracetamol (250 mg/144 g serving size)

The exact preparation of the formulation which will be consumed during a test day is described in the Investigational Nutritional Product Dossier. Table 13: Nutrition facts basic foam/liquid formulation

(Density: 1 .062 g/ml)

100 ml 100 g

Energy (kcal) 54 50.8

Total carbohydrate (g) 7.65 7.20

Sugars (g) 0.03 0.03

Lactose (g) 7.62 7.18

Total protein (g) 5.25 4.94

Total dietary fibre (g) 0.5 0.47

Table 14: Nutrition facts orange syrup

Per 100 ml Per 100 g

Energy 300 kcal 236.2 kcal

Protein 0.2 g 0.16 g

Carbohydrates 74 g 58.3 g

Sugars 72 g 56.7 g

Fat <0.1 g <0.08 g

Fibers 0.4 g 0.3 g

Sodium < 0.01 g < 8 mg

Vitamin B3 22 mg 17 mg

Vitamin B6 2.4 mg 1.9 mg

Vitamin C 72 mg 57 mg

Vitamin E 12 mg 9.4 mg

Table 15: Nutrition facts of the Paracetamol foam/liquid formulation

(Density: 1.062 g/ml)

100 ml 100 g

Energy (kcal) 64.6 69.3

Total carbohydrate (g) 1 1.5 12.3

Sugars (g) 1 1.8 12.6

Total protein (g) 4.2 4.5

Total dietary fibre (g) 0.42 0.45

Study procedure

Both products were consumed with a spoon and the start- and end time of consumption was recorded. The mean consumption duration of the liquid was 3.3 ± 1 .4 min and for the foam 5.5 ± 2.6 min and both mean consumption durations were within the instructed 10 min. Only one subject consumed the foam in 1 1 min. The maximum amount not consumed was 4 grams. The average paracetamol consumed was 249.7 mg (range 247.1 - 251 .8 mg) were target paracetamol dose was 250.0 mg.

Analysis

Plasma paracetamol concentrations were quantified using a rapid UPLC-MS/MS-based method developed at URDV. Paracetamol concentrations can be quantified within a range of 0.1 to 10 μΜ. For accuracy the following values were found: between 90-105% with an average of 98%.

At 10 time points five minutes after plasma sampling the subjects got a finger prick and a dried blood sampling (DBS) sample was taken. To allow calculation of the plasma concentration from the DBS samples, the hematocrit value was measured in these subjects as well.

Statistical

The primary study parameter was the Gastric Empying half time (GE50). Secondary study parameters were all parameters that describe the PK profile of paracetamol, e.g. area- under the plasma concentration time curve, absorption- and elimination rates.

Primary study parameter GE50 and all PK-parameters were analyzed using ANOVA model with treatment, visit and their interaction as fixed factors and subject as random factor and duration of consumption as covariate. The ANOVA results on those parameters were described by Least-Square mean (LSmean) and standard error (SE).

Pharmacokinetic evaluation

GE curves were constructed from the PK data based on Wagner-Nelson deconvolution method (Sanaka et al 1998, Nakada et al 1999). The following PK data were calculated 1 ) partial areas under the plasma concentrations time profile (AUC ^0" ') using non- compartmental analysis and 2) K _e i _{, l} iqui _d and total predicted area under the plasma concentration time curve (AUC° ^"∞ ) using compartmental analysis. The GE half time (GE50) was intrapolated from the individual curves with accuracy of 1 min or less.

WinNonLin version 6.2.1 was used for PK modelling. Results

Plasma paracetamol concentration time curves

After blind review of plasma data it was concluded that only one data set needed to be excluded for PP analysis due to increased baseline paracetamol value. The plasma concentrations were normalized to a body weight of 70 kg:

C70kg = (actual BW/70) measured.

In all cases (n=23) maximum plasma concentration were between 10 and 30 μΜοΙ/L. These maximum values were reached within 100 min post-dosing.

On an individual level the AUC's for 7 subjects (out of 1 1 , as for one subject a plasma paracetamol concentration was measured at T=0. Theoretically it should be 0, and it is presumed to be the result of a violation of the instructions not to consume painkillers during the study) are larger for foam compared to liquid and the Δ AUC's for those 7 subjects lie between 30 and 609 Γτιίη.μΜ/Ι_ (see table 15). Still, the average paracetamol plasma concentrations of both curves do overlap. Neither visual lag time, nor differences in apparent rate of absorption and elimination were observed. Foam had a slightly increased concentration at visual Tmax. Table 16: Individual AUC and GE50 values.

Subject # AUC Foam AUClJquid Delta AUCuquid-Foam

[ - ] [min ^* uM/L] [min ^* uM/L] [min ^* uM/L]

101 6256 6226 -30

102 3893 4148 255

104 4379 4315 -64

105 6096

106 5453 4957 -495

107 5129 5285 156

108 3241 3331 90

109 4658 4049 -609

1 10 3423 4092 668

1 1 1 5099 4902 -197

1 12 4436 4276 -160

1 13 3730 3187 -542 Subject # GE50,Foam GE ₅ o,|_iquid DeltaGE ₅ o,|_iquid-Foam

[ - ] [min] [min] [min]

101 19 28 9

102 15 17 2

104 24 16 -8

105 30

106 50 84 34

107 16 24 8

108 35 37 2

109 27 10 -17

1 10 28 24 -4

1 1 1 15 22 7

1 12 42 20 -22

1 13 28 45 17

Gastric emptying half time (GE50)

GE50 is not normal distributed and needed to be log transformed.

LS mean GE50 for foam is 26.9 min and for liquid 24.2 min (range respectively 19.6-36.9 min and 17.9-32.8 min) and descriptive GE50 for foam is 27.2 ± 1 1.4 min and for liquid 29.8 ± 19.5 min. There is no significant difference between the treatments on GE50.

Table 17 Individual Tmax and rate of absorption (Ka) values

Conclusion

The GE50 results show that at an almost equal mean (27.2 and 29.8 min), almost a factor 2 difference is obtained for the SD (1 1 .4 vs 19.5 min) in favour of the foam. This indicates a reduced variability of the foam stomach and small intestinal handling, further expressed in remarkably reduced SD values for rethe PK parameter describing absorption. For the foam treatment the SD value for the absorption rate (Ka) of paracetamol is with 1 1 versus 23 (min ^"1 ) about a factor 2 tighter, while the averages are close (24 and 33 (min ^"1 )). The effect on the reduced variability in absorption rate is even more clearly expressed in the ratio between maximum and minimum observations for Ka; 3.36 (foam) versus 13.02 (liquid). Tmax values show the same remarkable reduction in variability, where the ratio between maximum and minimum observation is 2.15 vs 4.74 in favour of the foam.

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Description of the figures

Figure 1. Stability of an aerated edible product (example 5) under simulated gastric conditions. Results are means ± standard deviation of 3 experiments.

Figure 2. Comparison of the behaviour of the aerated edible products in the stomach. The intensity scales are increased for better visualisation of the aerated edible product.

a) Stable Aerated edible product at time 10 minutes,

b) Stable Aerated edible product at time 50 minutes. These images are from the same volunteer on the same study day.

c) Less Stable Aerated edible product at time 10 minutes,

d) Less Stable Aerated edible product at time 50 minutes. Figure 3. Correlation between gastric emptying (expressed in volume) from the MRI study data (example 8) and the in-vitro test (example 9).

Legend:

• in-vitro stable foam

♦ in-vitro less stable foam

■ study less stable foam

▲ study stable foam

Figure 4. Linear regression between gastric emptying (expressed in volume) from the MRI study data (example 8) and the in-vitro test (example 9).

Legend:

♦ less stable foam

▲ stable foam