Oxyntomodulin for preventing or treating excess weight   

The present application is a continuation application of U.S. application Ser. No. 10/488,341, filed Nov. 8, 2004 (allowed), which is the U.S. National Phase of International Application No. PCT/GB02/04082, filed Sep. 9, 2002, which claims the benefit of British priority application no. UK 0121709.0, filed Sep. 7, 2001.

The present invention relates to compositions and methods for use in weight loss in mammalian animals.

One of the diseases with the highest incidence but which lacks effective treatment is obesity. It is a debilitating condition which reduces quality of life and substantially increases the risk of other diseases.

In the USA 25% of the adult population is now considered to be clinically obese. It has been estimated that $45 billion of US healthcare costs, or 8% per annum of total healthcare spending, is a direct result of obesity. In Europe the problem is increasing. It has been predicted that without new approaches over 20% of the UK population will be clinically obese by 2005. The fact that obesity is a metabolic disease is being increasingly recognised by the medical profession and the health authorities. There is, however, a shortage of effective and safe drugs which can be used in conjunction with diet and exercise for the long-term management of obesity.

It is an object of the present invention to provide such drugs and also to provide means to identify and develop such drugs.

Preproglucagon is a 160 amino acid polypeptide which is cleaved in a tissue specific manner by prohormone convertase-1 and -2 giving rise to a number of products with a variety of functions in both the central nervous system (CNS) and peripheral tissues. In the intestine and in the CNS, the major post-translational products of preproglucagon cleavage are glucagon-like peptide-1 (GLP-1), glucagon-like peptide-2 (GLP-2), glicentin and oxyntomodulin (OXM), as shown in Figure A. To date, no role in the CNS has been demonstrated for OXM.

While GLP-1 and GLP-2 have been shown to inhibit food intake, no such role has been demonstrated for the distinct peptide OXM. The importance of OXM as a biologically active peptide has not been demonstrated.

It has been surprisingly found that contrary to expectations, the OXM peptide can inhibit food intake and reduce weight.

Accordingly, the present invention provides, according to a first aspect, a composition comprising OXM, for use in the prevention or treatment of excess weight in a mammal.

In this text, the term “oxyntomodulin” is the same as “OXM” and relates to any composition which includes an OXM peptide sequence or an analogue thereof as follows:

OXM sequences are well known and documented in the art. The present invention relates to all of the sequences recited herein including, in particular, the OXM human sequence (which is the same as the rat, hamster and bovine OXM sequence), as follows:

(SEQ ID NO: 4) His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn Arg Asn Asn Ile Ala, the OXM angler fish sequence as follows:

(SEQ ID NO: 2) His Ser Glu Gly Thr Phe Ser Asn Asp Tyr Ser Lys Tyr Leu Glu Asp Arg Lys Ala Gln Glu Phe Val Arg Trp Leu Met Asn Asn Lys Arg Ser Gly Val Ala Glu, and the eel OXM sequence as follows:

(SEQ ID NO: 3) His Ser Gln Gly Thr Phe Thr Asn Asp Tyr Ser Lys Tyr Leu Glu Thr Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Ser Lys Arg Ser Gly Gly Pro Thr.

The term OXM used in this text also covers any analogue of the above OXM sequence, wherein the histidine residue at position 1 is maintained or replaced by an aromatic moiety carrying a positive charge or a derivative thereof, preferably wherein the moiety is an amino acid, more preferably wherein it is a histidine derivative, while 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 of the other amino acids in the above OXM sequence can be independently replaced by any other independently chosen amino acid, with the exception of histidine in position 1.

Any one or more (to 22) other alpha-amino acid residue in the sequence can be independently replaced by any other one alpha-amino acid residue. Preferably, any amino acid residue other than histidine is replaced with a conservative replacement as well known in the art i.e. replacing an amino acid with one of a similar chemical type such as replacing one hydrophobic amino acid with another.

As discussed above, 1 to 22 of the amino acids can be replaced. In addition to the replacement option above, this may be by a non-essential or modified or isomeric form of an amino acid. For example, 1 to 22 amino acids can be replaced by an isomeric form (for example a D-amino acid), or a modified amino acid, for example a nor-amino acid (such as norleucine or norvaline) or a non-essential amino acid (such as taurine). Furthermore, 1 to 22 amino acids may be replaced by a corresponding or different amino acid linked via its side chain (for example gamma-linked glutamic acid). For each of the replacements discussed above, the histidine residue at position 1 is unaltered or defined above.

In addition, 1, 2, 3, 4 or 5 of the amino acid residues can be removed from the OXM sequence with the exception of histidine at the 1 position (or as defined above). The deleted residues may be any 2, 3, 4 or 5 contiguous residues or entirely separate residues.

The C-terminus of the OXM sequence may be modified to add further amino acid residues or other moieties. The OXM above may be provided as the corresponding salt thereof. Examples of pharmaceutically acceptable salts of OXM and its analogues include those derived from organic acids such as methanesulphonic acid, benzenesulphonic acid and p-toluenesulphonic acid, mineral acids such as hydrochloric and sulphuric acid and the like, giving methanesulphonate, benzenesulphonate, p-toluenesulphonate, hydrochloride and sulphate, and the like, respectively or those derived from bases such as organic and inorganic bases. Examples of suitable inorganic bases for the formation of salts of compounds for this invention include the hydroxides, carbonates, and bicarbonates of ammonia, lithium, sodium, calcium, potassium, aluminium, iron, magnesium, zinc and the like. Salts can also be formed with suitable organic bases. Such bases suitable for the formation of pharmaceutically acceptable base addition salts with compounds of the present invention include organic bases which are nontoxic and strong enough to form salts. Such organic bases are already well known in the art and may include amino acids such as arginine and lysine, mono-, di-, or trihydroxyalkylamines such as mono-, di-, and triethanolamine, choline, mono-, di-, and trialkylamines, such as methylamine, dimethylamine, and trimethylamine, guanidine; N-methylglucosamine; N-methylpiperazine; morpholine; ethylenediamine; N-benzylphenethylamine; tris(hydroxymethyl)aminomethane; and the like.

Salts may be prepared in a conventional manner using methods well known in the art. Acid addition salts of said basic compounds may be prepared by dissolving the free base compounds in aqueous or aqueous alcohol solution or other suitable solvents containing the required acid. Where OXM contains an acidic function a base salt of said compound may be prepared by reacting said compound with a suitable base. The acid or base salt may separate directly or can be obtained by concentrating the solution eg. by evaporation. OXM may also exist in solvated or hydrated forms.

The OXM of the present invention may be conjugated to one or more groups such as a lipid, sugar, protein or polypeptide. The OXM can be conjugated by being attached to the group (for example via a covalent or ionic bond) or can be associated therewith. The conjugated link is preferably not through the C or N terminus amino acid, when the OXM is attached to the group. The OXM can be conjugated to a polymer such as polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, polyoxyethylene-polyoxypropylene copolymers, polysaccharides such as cellulose, cellulose derivatives, chitosan, acacia gum, karaya gum, guar gum, xanthan gum, tragacanth, alginic acid, carrageenan, agarose, and furcellarans, dextran, starch, starch derivatives, hyaluronic acid, polyesters, polyamides, polyanhydrides, and polyortho esters.

The OXM can be chemically modified. In particular, the amino acid side chains, the N terminus and/or the C acid terminus of OXM can be modified. For example, the OXM can undergo one or more of alkylation, disulphide formation, metal complexation, acylation, esterification, amidation, nitration, treatment with acid, treatment with base, oxidation or reduction. Methods for carrying out these processes are well known in the art. In particular the OXM is provided as a lower alkyl ester, a lower alkyl amide, a lower dialkyl amide, an acid addition salt, a carboxylate salt or an alkali addition salt thereof. In particular, the amino or carboxylic termini of the OXM may be derivatised by for example, esterification, amidation, acylation, oxidation or reduction. In particular, the carboxylic terminus of the OXM can be derivatised to form an amide moiety.

The OXM can be treated with metals, in particular with divalent metals. For the purposes of this invention the OXM can therefore be provided in the presence of one or more of the following metals, zinc, calcium, magnesium, copper, manganese, cobalt, molybdenum or iron.

The OXM can be provided in combination with a pharmaceutically acceptable carrier or diluent. Suitable carriers and/or diluents are well known in the art and include pharmaceutical grade starch, mannitol, lactose, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, (or other sugar), magnesium carbonate, gelatin, oil, alcohol, detergents, emulsifiers or water (preferably sterile). The composition may be a mixed preparation of a composition or may be a combined preparation for simultaneous, separate or sequential use (including administration). The OXM can be provided as a crystalline solid, a powder, an aqueous solution, a suspension or in oil.

The compositions according to the invention for use in the aforementioned indications may be administered by any convenient method, for example by oral (including by inhalation), parenteral, mucosal (e.g. buccal, sublingual, nasal), rectal, subcutaneous or transdermal administration and the compositions adapted accordingly.

For oral administration, the composition can be formulated as liquids or solids, for example solutions, syrups, suspensions or emulsions, tablets, capsules and lozenges.

A liquid formulation will generally consist of a suspension or solution of the compound or physiologically acceptable salt in a suitable aqueous or non-aqueous liquid carrier(s) for example water, ethanol, glycerine, polyethylene glycol or an oil. The formulation may also contain a suspending agent, preservative, flavouring or colouring agent.

A composition in the form of a tablet can be prepared using any suitable pharmaceutical carrier(s) routinely used for preparing solid formulations. Examples of such carriers include magnesium stearate, starch, lactose, sucrose and microcrystalline cellulose.

A composition in the form of a capsule can be prepared using routine encapsulation procedures. For example, powders, granules or pellets containing the active ingredient can be prepared using standard carriers and then filled into a hard gelatin capsule; alternatively, a dispersion or suspension can be prepared using any suitable pharmaceutical carrier(s), for example aqueous gums, celluloses, silicates or oils and the dispersion or suspension then filled into a soft gelatin capsule.

Compositions for oral administration may be designed to protect the active ingredient against degradation as it passes through the alimentary tract, for example by an outer coating of the formulation on a tablet or capsule.

Typical parenteral compositions consist of a solution or suspension of the compound or physiologically acceptable salt in a sterile aqueous or non-aqueous carrier or parenterally acceptable oil, for example polyethylene glycol, polyvinyl pyrrolidone, lecithin, arachis oil or sesame oil. Alternatively, the solution can be lyophilised and then reconstituted with a suitable solvent just prior to administration.

Compositions for nasal or oral administration may conveniently be formulated as aerosols, drops, gels and powders. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomising device. Alternatively the sealed container may be a unitary dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal once the contents of the container have been exhausted. Where the dosage form comprises an aerosol dispenser, it will contain a pharmaceutically acceptable propellant. The aerosol dosage forms can also take the form of a pump-atomiser.

Compositions suitable for buccal or sublingual administration include tablets, lozenges and pastilles, wherein the active ingredient is formulated with a carrier such as sugar and acacia, tragacanth, or gelatin and glycerin.

Compositions for rectal or vaginal administration are conveniently in the form of suppositories (containing a conventional suppository base such as cocoa butter), pessaries, vaginal tabs, foams or enemas.

Compositions suitable for transdermal administration include ointments, gels, patches and injections including powder injections.

Conveniently the composition is in unit dose form such as a tablet, capsule or ampoule.

The OXM can be used as a prophylaxis to prevent excess weight gain or can be used as a therapeutic to lose excess weight.

The excess weight is typically obesity, although the mammal will not be certified as clinically obese in order to be suffering from excess weight. The OXM may be in liquid, solid or semi-solid form.

In today\'s society, the prevention or treatment of excess weight in a mammal is a real need. Preferably the mammal is a human, although it may also include other mammalian animals, such as horses, canine animals (in particular domestic canine animals), feline animals (in particular domestic feline animals) as well as mammals which are produced for meat, such as porcine, bovine and ovine animals. The present invention can be used to prevent excess weight in such animals in order to maximise lean meat production.

Throughout this text, the term “prevention” means any effect which mitigates any excess weight, to any extent. Throughout this text, the term “treatment” means amelioration of excess weight, to any extent.

According to a second aspect, the present invention provides a method for the prevention or treatment of excess weight in a mammal, the method comprising administering a composition comprising OXM to a mammal. The mammal is likely to be in need of prevention or treatment of excess weight. The weight loss may be cosmetic. The composition comprising OXM will be administered in an effective concentration.

All preferred features of the first aspect of the invention, also apply to the second.

A third aspect of the present invention provides a method for cosmetic weight loss in a mammal, the method comprising administering a composition comprising OXM to a mammal. In this circumstance, the weight loss is purely for the purposes of cosmetic appearance.

All preferred features of the first and second aspects also apply to the third.

Without being bound to this theory, it is understood that the present invention provides the prevention or treatment of excess weight by the administration of OXM which acts as an inhibitor to food intake to the mammalian body. Such reduced food intake results in the prevention or treatment of excess weight in a mammal. In this text the term “food” includes a substance which is ingested and which has calorific value.

A fourth aspect to the present invention provides the use of OXM in the manufacture of a medicament for the prevention or treatment of excess weight.

All preferred features of the first and third aspects, all apply to the fourth.

The first, second and fourth aspects of the invention relate to medicaments, the particular dosage regime for which will ultimately be determined by the attending physician and will take into consideration such factors as the OXM being used, animal type, age, weight, severity of symptoms and/or severity of treatment to be applied, method of administration of the medicament, adverse reaction and/or contra indications. Specific defined dosage ranges can be determined by standard designed clinical trials with patient progress and recovery being fully monitored.

Such trials may use an escalating dose design using a low percentage of the maximum tolerated dose in animals as the starting dose in man.

The fifth aspect of the present invention relates to the use of OXM to identify an agent which inhibits food intake in a mammal. This aspect of the invention provides a means to identify and develop further suitable medicaments for the prevention or treatment of excess weight.

The use of OXM may include the use of the peptide itself or may include the use of the theoretical or models characteristics of OXM. The functional or structural characteristics of OXM utilised may be of a peptide itself or may be of a computer-generated model, a physical two or three-dimensional model or an electrical generated (e.g. computer generated) primary, secondary or tertiary structure, as well as the pharmacophore (three-dimensional electron density map) or its X-ray crystal structure.

The structural characteristics may enable the identification of potential agents that may interact with OXM thereby affecting it\'s function. The identification may be by computer modelling and/or rational drug design.

Preferred features of each aspect of the invention are as for each of the other aspects mutatis mutandis.

The present invention is now described by way of example only and with reference to the following figures, in which:

FIG. A is a graphical representation of preproglucagon and its component parts;

FIG. 1 is a comparison of the effects of ICV and iPVN proglucagon-derived and related products on food intake in fasted rats. FIG. 1A illustrates the cumulative food intake (g) up to 8 h after ICV injection of GLP-1, OXM, glucagon, or glicentin (all 3 nmol) into fasted animals. *, P<0.05 vs. saline control. FIG. 1B illustrates cumulative food intake (g) up to 24 h after an acute iPVN injection of GLP-1, OXM (both 1 nmol), or exendin-4 (0.03 nmol) into fasted animals. *, P<0.01 vs. saline control for all groups at 1, 2, and 4 h. *, P<0.05 vs. saline control for exendin-4 only at 8 h;

FIG. 2 shows two graphs of the effects of ICV and iPVN OXM on food intake in fasted rats. FIG. 2A, cumulative food intake (g) up to 8 h after an acute ICV injection of OXM (0.3, 1, 3, or 10 nmol). FIG. 2B, cumulative food intake (g) up to 8 h after an acute iPVN injection of OXM (0.1, 0.3, or 1.0 nmol) into fasted animals. *, P<0.05 vs. saline control;

FIG. 3 shows two bar graphs of the effect of ICV OXM at the onset of the dark phase. Sated rats received an ICV injection of OXM, GLP-1 (3 nmol), or saline at the onset of the dark phase. Food intake (grams; A) and behaviors (B) at 1 h postinjection were determined. *, P<0/05 vs. saline control;

FIG. 4 shows two bar graphs of the inhibition of OXM and GLP-1 effects on food intake by exendin-(9-39). FIG. 4A, food intake 1 h after an acute ICV injection of GLP-1 (3 nmol), GLP-1 plus exendin-(9-39) (30 nmol), OXM (3 nmol), OXM and exendin-(9-39) (30 nmol), or exendin-(9-39) alone (30 nmol). FIG. 4B, food intake after an acute iPVN injection of GLP-1 (1 nmol), GLP-1 and exendin-(9-39) (10 nmol), OXM (1 nmol), OXM and exendin-(9-39) (10 nmol), or exendin-(9-39) alone (10 nmol) into fasted animals. **, P<0.005 vs. saline control;

FIG. 5 is a graph of the competition of [125I] GLP-1 binding in rat hypothalamic membranes by GLP-1 and OXM;

FIG. 6 illustrates the effect of a) IP OXM (30, 100 and 300 nmol/kg in 500 μl saline) or saline on cumulative food intake (g) in 24-hour fasted rats injected during the early dark phase (closed squares=saline, open circles=OXM 30 nmol/kg, closed triangles=OXM 100 nmol/kg, open triangles=OXM 300 nmol/kg); and b) IP OXM (30 and 100 nmol/kg in 500 μl saline) or saline on cumulative food intake in non-fasted rats injected prior to the onset of the dark phase (closed squares=saline, open circles=OXM 30 nmol/kg, closed triangles=OXM 100 nmol/kg). *P<0.05 vs. saline;

FIG. 7 illustrates the effect of twice daily IP injections of OXM (50 nmol/kg) or saline for seven days on a) cumulative food intake (g); and b) body weight gain (g). *P<0.05, **P<0.01, ***P<0.005 vs. saline;

FIG. 8 illustrates the effect of IP OXM (50 nmol/kg), saline or a positive control (1 hour=GLP-1 (50 nmol/kg); 2 hours=CCK (15 nmol/kg)) on gastric emptying in 36-hour fasted rats. Contents (dry weight) of the stomach were expressed as a percentage of the food intake during the 30-minute feeding period. **P<0.01 vs. saline;

FIG. 9 illustrates the effect of increasing doses of OXM (0.01-1.0 nmole) on 1 hour food intake when administered into the arcuate nucleus of 24-hour fasted rats. *P<0.05, **P<0.01, ***P<0.05 vs. saline;

FIG. 10 illustrates the effect of iARC administration of exendin 9-39 (5 nmoles) or saline injected 15 minutes prior to IP administration of OXM (30 nmol/kg), GLP-1 (30 nmol/kg) or saline on 1 hour food intake (g). (S=saline, G=GLP-1 (30 nmol/kg), Ox=OXM (30 nmol/kg), Ex=exendin 9-39 (5 nmoles));

FIG. 11a illustrates the expression of fos-like immunoreactivity in response to A) IP saline or B) IP OXM (50 nmol/kg) in the arcuate nucleus of the hypothalamus (×40 magnification). ***P<0.005 vs. saline; and

FIG. 11b illustrates the expression of fos-like immunoreactivity in response to A) IP saline, B) IP OXM (50 nmol/kg) or C) IP CCK (15 nmol/kg) in the NTS and AP of the brainstem.

GLP-1, glicentin, glucagon, and SP-1 were purchased from Peninsula Laboratories, Inc. (St. Helens, UK). OXM was purchased from IAF BioChem Pharma (Laval, Canada). Exendin-4 and exendin-(9-39) were synthesised at Medical Research Council, Hemostasis Unit, Clinical Sciences Center, Hammersmith Hospital, London, UK using F-moc chemistry on an 396 MPS peptide synthesiser (Advanced ChemTech, Inc.) and purified by reverse phase HPLC on a C8 column (Phenomex, Macclesfield, UK). The correct molecular weight was confirmed by mass spectrometry. All chemicals were purchases from Merck & Co. (Lutterworth, Leicester, UK) unless otherwise stated.

Adult male Wistar rats (ICSM, Hammersmith Hospital) were maintained in individual cages under controlled conditions of temperature (21-23° C.) and light (12 h of light, 12 h of darkness) with ad libitum access to food (RM1 diet, Special Diet Services UK Ltd., Witham, UK) and tap water. Animals were handled daily after recovery from surgery until completion of the studies. All animal procedures undertaken were approved by the British Home Office Animals (Scientific Procedures Act 1986 (Project License PIL 90/1077).

ICV and iPVN Cannulation and Infusions of Test Compounds

Animals had permanent stainless steel guide cannulas (Plastics One, Roanoke, Va.) stereotactically implanted ICV or iPVN. All studies were carried out in the early light phase, between 0900-1100 h, after a 24-h fast, and food intake was measured 1, 2, 4, 8, and 24 h postinjection.

Comparison of the effect of proglucagon-derived products and related peptides on food intake.

In study 1a, rats were injected ICV with 10 μl saline, GLP-1 (13 nmol), OXM (3 nmol), glucagon (3 nmol), or glicentin (3 nmol; n=8/group).

In all studies, the OXM with the following sequence was used:

(SEQ ID NO: 1) His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg Ala Gln

Download full PDF for full patent description/claims.




You can also Monitor Keywords and Search for tracking patents relating to this Oxyntomodulin for preventing or treating excess weight patent application.

Patent Applications in related categories:

20130116173 - Glucagon analogs exhibiting gip receptor activity - Provided herein are glucagon analogs which exhibit potent activity at the GIP receptor, and, as such are contemplated for use in treating diabetes and obesity. In exemplary embodiments, the glucagon analog of the present disclosures exhibit an EC50 at the GIP receptor which is within the nanomolar or picomolar range. ...

20130116172 - Glucagon superfamily peptides exhbiting g protein coupled receptor activity - Provided herein are glucagon superfamily peptides conjugated with GPCR ligands that are capable of acting at a G protein-coupled receptor. Also provided herein are pharmaceutical compositions and kits of the conjugates of the invention. Further provided herein are methods of treating a disease, e.g., a metabolic disorder, such as diabetes ...


###


Other recent patent applications listed under the agent Lando & Anastasi, LLP:

20090322790 - System and method for streamlining user interaction with electronic content
20090322953 - Method and apparatus for achieving selected audio/video and other functions
20090323258 - Apparatus and method for scalable power distribution
20090325311 - Measurement methods
20090327141 - Highly efficient secrecy-preserving proofs of correctness of computation
20090314122 - Apparatus for automatically adjusting tension of cable in parking brake and parking brake apparatus using the apparatus
20090314354 - Method and apparatus for monitoring and controlling pressure in an inflatable device
20090317554 - Apparatus and method for spray coating
20090318527 - Indole compounds and methods of use thereof
20090319356 - Systems and methods for managing electronically delivered information channels
20090307961 - Animal trap having timed release door
20090308579 - Exhaust air removal system
20090309570 - Methods and systems for managing facility power and cooling
20090311183 - Metalloproteinase 12 binding proteins
20090311245 - Metalloproteinase 9 binding proteins
20090300846 - Inflatable device forming mattresses and cushions
20090301324 - Method and apparatus for placing substrate support components
20090303676 - System and method for streamlining user interaction with electronic content
20090307337 - Apparatus and method for device identification
20090294610 - Picture hanger assembly and method
20090295594 - Demand response method and system
20090297449 - Metalloproteinase 9 and metalloproteinase 2 binding proteins
20090298157 - Heparan sulfate glycosaminoglycan lyase and uses thereof
20090300511 - System and method for streamlining user interaction with electronic content