WO2016035052A9

WO2016035052A9 - Anti-nerve growth factor antibodies for treating itching

Info

Publication number: WO2016035052A9
Application number: PCT/IB2015/056771
Authority: WO
Inventors: David Gearing; B. Duncan X. LASCELLES
Original assignee: Nexvet Australia Pty Ltd
Priority date: 2014-09-05
Filing date: 2015-09-04
Publication date: 2016-09-22
Also published as: WO2016035052A1

Abstract

Methods of treating itching using NGF antagonists are provided, as are NGF antagonists and NGF antagonist-containing compositions for use in such methods.

Description

ANTI-NERVE GROWTH FACTOR ANTIBODIES FOR TREATING ITCHING

Cross Reference to Related Applications

This application claims the benefits of priority to U.S. Provisional Application No.

62/046,428, filed September 5, 2014, the entire contents of which are incorporated herein by reference.

Field of the Invention

The present invention relates to the use of NGF antagonists, such as anti-NGF antibodies, NGF-binding antibody fragments, NGF-binding proteins or polypeptides, or small molecule chemicals, for treating itching, and to compositions comprising an NGF antagonist for use in such methods, and to related uses of NGF antagonists.

Background

Itching and chronic itching is a major problem in humans and animals, including horses, dogs, and cats. It has been estimated that at least 1/3 of all Americans suffer from itching (see, e.g., Bickers et al., JAAD 55:490-500 (2004); Thorpe et al., Health Aff. 22: W4-437-445 (2004)) and that economic costs associated with chronic itching (such as that associated with atopic dermatitis) amount to about $3 billion annually. It has been estimated that atopic dermatitis affects 10-15% of canines. See, e.g., Scott & Paradis, Can. Vet. J. 31 : 830-35 (1990); Budgin & Flaherty, Vet. Clin. Small Anim. 43: 189-204 (2013). A number of agents that treat itching are known, include antihistamines, 5-HT3 antagonists, corticosteriods, μ-opioid antagonists (Naltrexone), k-opioid antagonists and anti-depressants (including SSRIs such as paroxetine and sertraline), and other agents are in development, such as anti-IL-31 , neurokinin-1-receptor antagonists, and H4- receptor antagonists. See, e.g., Raap et al., Curr. Op. Allergy & Clin. Immunol. 11 : 420- 27 (201 1). However, many of these agents have not been useful against itching associated with specific conditions, such as atopic dermatitis, or kidney or liver disease.

Nerve growth factor (NGF) is a naturally occurring secreted protein which consists of an alpha, beta and gamma polypeptide chain. NGF is a member of the neurotrophin family and is implicated in a number of different roles. NGF promotes survival and differentiation of sensory and sympathetic neurons and signals via two membrane bound receptors, p75, a low affinity NGF receptor and TrkA, a transmembrane tyrosine kinase and a high affinity NGF receptor. The binding of NGF to TrkA or p75 results in an upregulation of neuropeptides in sensory neurons. The use of NGF antagonists to treat pain and pain sensitivity in humans has been described. See, e.g., Cattaneo, Curr. Op. Mol. Ther. 12:94-106 (2010). However, while pain and itching share some common pathways, they differ in a number of respects. See, e.g., Akiyama & Carstens, Neurosci. 250: 697-714 (2013); Han et al., Nature Neurosci. 16: 174-84 (2013). Notably, while μ-opioids attenuate pain, they can exacerbate itching. See, e.g., Akiyama & Carstens, supra. Moreover, painful stimuli can inhibit itching. Id. Recently, itch-specific neurons have been identified that are linked to itching but not pain (Han et al., supra), and distinct animal models have been developed for pain and itch (Akiyama & Carstens, supra).

Although chronic itching has been associated with increased levels of NGF (see, e.g., Rukwied et al., J. Invest. Derm. 133: 268-71 (2013)) and NGF is up-regulated in human atopic dermatitis lesions and itchy cutaneous tumors (see, e.g., Suga etal., Acta Derm. Venereol. 93: 144-49 (2013)), NGF antagonists have not heretofore been shown to be effective in treating itching. Takano et al., J. Pharmacol. Sci. 99: 277-86 (2005), investigated the effects of anti-NGF antibody on symptoms in a murine model of atopic dermatitis, and found that the anti-NGF antibodies inhibited the development and proliferation of skin lesions and epidermal innervation, and inhibited growth in itching, but did not ameliorate scratching already developed. Rukwied et al., supra, reported that NGF sensitized nociceptors for cowhage-induced itch, but not histamine-induced itch in human skin. Indeed, the current understanding of itching is that itch transduction can occur via different mediators and mechanisms. For example, histamine, cowhage, and IL-31 have been identified as peripheral itch mediators. See, e.g., Carstens, "Pruritus: Current Understanding of its Underlying Mechanisms," Veterinary Pain Short Course, Univ. of California, Davis (June 21 , 2014). Moreover, acute itching and chronic itching may involve different mechanisms. Id. Thus, there remains a need for effective methods of treating itching. Summary

In accordance with some embodiments, there are provided methods of treating itching in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an NGF antagonist In some embodiments, the subject is human, equine, canine or feline. In some embodiments,the itching is chronic itching or acute itching, and may be associated with one or more conditions selected from the group consisting of skin diseases, skin conditions, contact skin reactions, cancer, kidney disease, liver disease, viral infections, parasites, food allergies, and drug reactions. In some embodiments, the itching is associated with atopic dermatitis or is idiopathic itching.

For any subject and type of itching being treated, the NGF antagonist may be an anti- NGF antibody or NGF-binding antibody fragment In some embodiments, the subject is human and the NGF antagonist is a humanized anti-NGF antibody or humanized NGF- binding antibody fragment, such as tanezumab. In some embodiments, the subject is equine and (he NGF antagonist is an equinized anti-NGF antibody or equinized NGF- binding antibody fragment In some embodiments, the subject is canine and the NGF antagonist is a caninized anti-NGF antibody or caninized NGF-binding antibody fragment. In some embodiments, the subject is feline and the NGF antagonist is a feltnized anti-NGF antibody or felinized NGF-binding antibody fragment

For any subject and type of itching being treated, the NGF antagonist may be an NGF- binding polypeptide, such as polypeptides comprising TrkA and polypeptides comprising p75, wherein the polypeptides optionally further comprise an immunoglobulin Fc region.

For any subject, any type of itching being treated, and any type of NGF antagonist the NGF antagonist may be administered in a pharmaceutical composition. In some embodiments, the NGF antagonist Is administered parenterally, such as intravenously or subcutaneously.

In some embodiments.the subject is suffering from chronic itching and the NGF antagonist is administered once every four to six weeks.

For any subject, any type of itching being treated, and any type of NGF antagonist, the- method may further comprise administering an additional agent useful against itching. For any subject, any type of itching being treated, and any type of NGF antagonist, the subject may not be receiving steroid treatment, such as corticosteroid treatment.

In some embodiments, there is provided an NGF antagonist for use in a method of treating itching in a subject in need thereof, in accordance with the various embodiments described herein.

In some embodiments, there is provided the use of an NGF antagonist in the

manufacture of a medicament for use in a method of treating itching in a subject in need thereof, as described herein.

Brief Description of the Drawings FIG. 1 illustrates a pruritus visual analog scale (PVAS) used to score symptoms of canine atopic dermatitis. Users cross-mark a 10 cm line at the point best describing a dog's condition, and numerical scores are generated based on the distance of the mark (in cm) from the "normal dog" baseline.

FIG. 2 shows the change in PVAS score of each dog in the placebo group from baseline (day 0) at time points 7, 14, 21 , 28, and 42 days post-treatment (a negative value indicates improvement).

FIG. 3 shows the change in PVAS score of each dog in the NV-01 treated group from baseline (day 0) at time points 7, 14, 21 , 28, and 42 days post-treatment (a negative value indicates improvement). Detailed Description

Described herein are methods of treating itching, including chronic itching, using NGF antagonists, such as anti-NGF antibodies and NGF-binding antibody fragments or NGF- binding proteins or polypeptides, or small molecule chemicals.

Definitions Technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which the present invention pertains, unless otherwise defined. Reference is made herein to various methodologies known to those of ordinary skill in the art. Publications and other materials setting forth such known methodologies to which reference is made are incorporated herein by reference in their entireties as though set forth in full. Any suitable materials and/or methods known to those of ordinary skill in the art can be utilized in carrying out the present invention. However, specific materials and methods are described. Materials, reagents and the like to which reference is made in the following description and examples are obtainable from commercial sources, unless otherwise noted.

As used herein, the singular forms "a," "an," and "the" designate both the singular and the plural, unless expressly stated to designate the singular only.

The term "about" means that the number comprehended is not limited to the exact number set forth. As will be understood by persons of ordinary skill in the art, the meaning of "about" will vary to some extent on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art given the context in which it is used, "about" will mean up to plus or minus 10% of the particular value. As used herein "subject" denotes any animal, including human, equine, canine, or feline. In some embodiments, the subject is in need of treatment or prevention of itching, such as suffering from or being at risk of developing itching, including chronic itching or pruritus.

As used herein, the terms "itching" and "pruritus" are used interchangeably in

accordance with their art-recognized meaning to refer to "an unpleasant cutaneous sensation which provokes the desire to scratch." See, e.g., Akiyama & Carstens, Neuroscience 250: 697-714 (2013). Generally, itching lasting more than 6 weeks is termed "chronic" pruritus. See, e.g., Raap et al., Curr. Op. Allergy & Clin. Immunol. 1 1 : 420-27 (2011). As used herein, the phrase "therapeutically effective amount" means that dose or amount that provides the specific pharmacological response for which the agent is administered in the subject at hand, such as alleviation of itching.

As used herein, the term "NGF antagonist" means an agent that is capable of neutralising the biological activation and signalling of NGF. For example, the NGF antagonist may inhibit the binding of NGF to its target ligand, such as the cell membrane bound TrkA or p75 receptors. In some embodiments, the NGF antagonist is an anti- NGF antibody or NGF-binding antibody fragment or NGF-binding protein or polypeptide that specifically, and optionally selectively, binds to NGF and inhibits one or more biological activities of NGF. As used herein, the term "biological activity" refers to any one or more inherent biological properties of a molecule (whether present naturally as found in vivo, or provided or enabled by recombinant means). Biological properties include but are not limited to receptor binding and/or activation; induction of cell signalling or cell proliferation, inhibiting cell growth, induction of cytokine production, induction of apoptosis, and enzymatic activity.

The terms "polypeptide", "peptide", or "protein" are used interchangeably herein to designate a linear series of amino acid residues connected one to the other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues. The amino acid residues are usually in the natural "L" isomeric form. However, residues in the "D" isomeric form can be substituted for any L-amino acid residue, as long as the desired functional property is retained by the polypeptide.

As herein defined an "antibody" encompasses antigen-binding proteins which specifically bind to a target antigen of interest, in this case NGF, comprised of one or more polypeptides that can be recombinantly prepared or which are genetically encodable by immunoglobulin genes, or fragments of immunoglobulin genes. The term "antibody" encompasses monoclonal and chimeric antibodies, and further encompasses polyclonal antibodies and antibodies of any class or subtype. An "antibody" further extends to hybrid antibodies, and bispecific antibodies, heteroantibodies which retain antigen binding. The phrase "specifically binds to" refers to binding to a specific protein or target which is present amongst a heterogeneous population of proteins. Hence, when present in specific immunoassay conditions, the antibodies (for example) bind to a particular protein, in this case NGF, and do not bind in a significant amount to other proteins present in the sample. NGF Antagonists

The methods described herein use NGF antagonists. As set forth above, an NGF antagonist is any agent that is capable of neutralising the biological activation and signalling of NGF. For example, the NGF antagonist may inhibit the binding of NGF to its target ligand, such as the cell membrane bound TrkA or p75 receptors. In some embodiments, the NGF antagonist is an anti-NGF antibody, NGF-binding antibody fragment or NGF-binding protein or polypeptide.

NGF antagonists, including anti-NGF antibodies, NGF-binding antibody fragments and NGF-binding proteins or polypeptides are known. In some embodiments, the NGF antagonist is designed for use in the specific species being treated. For example, when the subject is human, a human or humanized NGF antagonist may be used, such as RN624 (tanezumab), JNJ-42160443, REGN475, PG1 10, alpha-D11 , AMG-403 (see, e.g., Mantyh et al., Anaesthesiology 115: 189-204 (2011)), the 91 1 antibody described in Hongo et al., Hybhdoma: ; 19: 215-27 (2000), or the aD1 1 antibody described in Ruberti et al., Cell. Molec. Neurobiol. 13: 559-68 (1993), or any of those described in WO 2012/153121 ; WO 2012/153122; WO 2012/153123; WO 2012/153126. In some embodiments, the human or humanized NGF antagonist is any one of those described in WO 2013/034900. When the subject is equine, an equine or equinized NGF antagonist may be used, such as any of those described in WO 2012/153123; when the subject is canine, a canine or caninized NGF antagonist may be used, such as any of those described in WO 2012/153121 , including NV-01 described in the examples below; when the subject is feline, a feline or felinized NGF antagonist may be used, such as any of those described in WO 2012/153122.

In some embodiments, the NGF antagonist is an NGF-binding protein or polypeptide, such as a polypeptide comprising TrkA or p75, or a fragment of TrkA or p75 that binds NGF. In specific embodiments, the NGF antagonist is an NGF-binding polypeptide comprising TrkA or p75 (or an NGF-binding fragment thereof) and an immunoglobulin Fc region. In some embodiments, the NGF antagonist is a trkA-lgG fusion protein that binds to NGF, such as described in Shelton et al., J. Neurosci. 15: 477-91 (1995). In some embodiments, the NGF antagonist is a p75 fusion protein that binds to NGF, such as p75-Fc being developed by Levicept (Kent, England) for the treatment of chronic pain. In some embodiments, the NGF antagonist is a small molecule chemical compound that interferes with the binding of NGF to TrkA, such as ALE0540 and PD90780, as described in Hefti et al., TRENDS in Pharmacol. Sci. 27: 85-91 (2006), or MNAC13, as described in Mantyh et al., Anaesthesiology 115: 189-204 (2011). In some

embodiments, the NGF antagonist is a small molecule chemical compound that inhibits TrkA.

Polypeptides (e.g., antibodies, antibody fragments and proteins) for use in the methods described herein may be produced by recombinant means, directly or as a fusion polypeptide with a heterologous polypeptide, such as a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide. The heterologous signal sequence may be selected to be recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For prokaryotic host cells that do not recognize and process a native antibody signal sequence, the signal sequence may be substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.

The term "isolated" when used in reference to polypeptides refers to the state in which said polypeptides are isolated from a natural source and/ or in purified form, such as having been separated, isolated or purified from their natural environment. Accordingly, such isolated polypeptides will be free or substantially free of material with which they are naturally associated, such as other polypeptides or nucleic acids with which they are found in their natural environment, or the environment in which they are prepared (e.g., cell culture) when such preparation is by recombinant DNA technology practiced in vitro or in vivo. The antibodies, fragments and proteins may be glycosylated, either naturally or by systems of heterologous eukaryotic cells (e.g. CHO or NSO cells, or they may be (for example if produced by expression in a prokaryotic cell) unglycosylated.

In some embodiments, the methods use a heterogeneous preparation of NGF antagonists comprising different NGF antagonists, such as one or more anti-NGF antibodies, one or more NGF-binding antibody fragments and/or one or more NGF- binding proteins or polypeptides. Methods and Pharmaceutical Compositions

Described herein are methods of treating itching, including chronic itching, using NGF antagonists, such as anti-NGF antibodies or NGF-binding antibody fragments or NGF- binding proteins or polypeptides. In some embodiments, the methods use a pharmaceutical composition comprising an NGF antagonist, optionally in a pharmaceutically acceptable carrier, excipient or diluent. Examples of suitable pharmaceutical carriers include; water, glycerol, ethanol and the like. The pharmaceutical composition may also include one or more preservatives, stabilizers, buffers, antioxidants and/or other additives typically used in pharmaceutical compositions, which may depend on the route of administration, target subject (e.g., human vs. equine), and anticipated length of storage.

The composition may be formulated for any route of administration. In specific embodiments, the composition is formulated for parenteral administration, such as intravenous infusion or injection. Other routes for parenteral administration include, but are not limited to intracardial, intraarterial, intraperitoneal, intramuscular, intracavity, subcutaneous, transmucosal, by inhalation or transdermally. Additional routes of administration include topical and enteral, for example, mucosal (including pulmonary), oral, nasal, and rectal.

In some embodiments, the pharmaceutical composition is a liquid formulation, a lyophilized formulation, a lyophilized formulation that is reconstituted as a liquid prior to use, or an aerosol formulation. The composition may also be administered via microspheres, liposomes, other microparticulate delivery systems or sustained release formulations placed in certain tissues including blood.

In embodiments where the composition is administered as an injectable composition, for example for intravenous, intradermal or subcutaneous administration, the NGF antagonist can be formulated in a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability. Those of relevant skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles such as sodium chloride injection, Ringer's injection or, Lactated Ringer's injection. Such compositions may also include one or more preservatives, stabilizers, buffers, antioxidants and/or other additives. In some embodiments, the composition further comprises a buffer. In some

embodiments, the buffer is a phosphate buffer, such as a composition comprising phosphate-buffered saline. In certain embodiments, the buffer comprises sodium chloride at a concentration of from about 10mM to about 300mM, such as about 140mM, and a concentration of phosphate of about 5 mM to about 20 mM, such as about 10 mM. In further specific embodiments, such a composition has a pH of about 7 to about 7.5, such as a pH of about 7.4. In some embodiments, the composition comprises sorbitol, such as a composition comprising soduium chloride, phosphate, and sorbitol. In further specific embodiments, such a composition has a pH of about 6.7 to about 7.0. In other embodiments, the buffer may be histidine buffer (about 4 mM to about 60 mM), succinate buffer (about 5 mM to about 25 mM), or acetate buffer (about 5 mM to 25 mM).

In some embodiments the composition further comprises a surfactant, optionally at a concentration of just above 0% to about 0.2%. The surfactant may be any surfactant suitable for use in the pharmaceutical composition, such as may be selected from the group consisting of polysorbate-20, polysorbate-40, polysorbate-60, polysorbate-65, polysorbate-80, polysorbate-85, and combinations thereof. In specific embodiments, the surfactant is polysorbate-20.

In some embodiments, the NGF antagonist is present in the composition at a concentration of about 0.5 mg/ml to about 250 mg/ml, about 0.5 mg/ml to about 100 mg/ml, about 0.5 mg/ml to about 45 mg/ml, about 100 mg/ml to about 200 mg/ml, or about 50 mg/ml to about 250 mg/ml. For example, when the NGF antagonist is a monoclonal antibody, it may be present in the composition at a concentration of about 0.5 mg/ml to about 250 mg/ml, including at about 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg/ml. Examples of the techniques and protocols mentioned above and other techniques and protocols which may be used in accordance with formulating compositions for use in the methods described herein can be found in Remington's Pharmaceutical Sciences, 18th edition, Gennaro, A.R., Lippincott Williams & Wilkins; 20th edition ISBN 0-912734-04-3 and Pharmaceutical Dosage Forms and Drug Delivery Systems; Ansel, H.C. et al. 7th Edition ISBN 0-683305-72-7, the entire disclosures of which are herein incorporated by reference. The NGF antagonist generally is administered to a subject in need thereof, such as a subject suffering from or at risk of developing itching. Pruritus (itching) may be associated with a number of different conditions, such as skin diseases (e.g., psoriasis or atopic dermatitis), skin conditions (e.g., aging, dry skin, burns), contact skin reactions (such as poison ivy), and other conditions, such as cancers, kidney disease, liver disease, viral infections (Herpes zoster) parasites, food allergies, drug reactions (such as reactions to spinal opiates), psychiatric conditions, and unknown reasons (e.g., idiopathic itching). Thus, in some embodiments, a subject may be suffering from or at risk of developing one or more of these conditions, or may be suffering from or at risk of developing itching associated with one or more of these conditions. In some

embodiments, the subject is suffering from chronic itching (e.g., itching that has lasted for at least 6 weeks). In some embodiments, the subject is suffering from acute itching.

The NGF antagonist generally is administered to a subject in a "therapeutically effective amount," this being an amount sufficient to show benefit to the subject to whom the composition is administered, such as alleviation of itching, such as reduction or elimination of itching. The actual dose administered, and rate and time-course of administration, will depend on, and can be determined with due reference to, the nature and severity of the condition which is being treated, as well as factors such as the age, sex and weight of the subject being treated, as well as the route of administration.

Further due consideration may be given to the properties of the composition, for example, the binding activity and in vivo plasma life of the NGF antagonist, the concentration of the NGF antagonist in the composition, as well as the route, site and rate of delivery.

Dosage regimens can include a single administration of the NGF antagonist

composition, or multiple administrations of the composition. Multiple adminsitrations can be several times a day, once a day, once every 2-3 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once a month, once every 6 weeks, once every 4-6 weeks, once every 2 months, one every 6 months, or as needed. In some embodiments, the NGF antagonist is administered not more often than once a month, not more often than once every 4-6 weeks, not more often than once every 6 weeks, not more often than once every 8 weeks, or less frequently. In general, an NGF antagonist that is a polypeptide or antibody fragment may require more frequent dosing than an NGF antagonist that is an antibody. Examples of suitable doses include from 1 μg/kg to 20 mg/kg, 1 μg/kg to 10mg/kg, 10 μg/kg to 1 mg/kg, based on the amount of NGF antagonist and weight of the subject.

In certain embodiments, the NGF antagonist is an anti-NGF antibody or NGF-binding antibody fragment or NGF-binding protein, and is administered at a dose ranging from about 0.01 mg/kg of body weight to about 30 mg/kg of body weight, including from 0.01 mg/kg of body weight to about 10 mg/kg of body weight, including from 0.03 mg/kg of body weight to about 3 mg/kg of body weight, including 0.02 mg/kg of body weight to about 2 mg/kg of body weight. For example, when the NGF antagonist is a monoclonal antibody, it may be administered at a dose of from about 0.01 mg/kg of body weight to about 30 mg/kg of body weight, including a dose of about 0.05, 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, or 2.0 mg/kg body weight. In canines, a typical dose may range from about 0.2 to about 2 mg/kg

In certain embodiments, the methods further comprise administering a further agent which may enhance and/or complement the effectiveness of the NGF antagonist, such as an antihistamine, analgesic, or steroid. In such embodiments, the other agent(s) may be administered as part of the same composition as the NGF antagonist, or may be administered in a separate composition, in which case it may be administered simultaneously or sequentially (before or after administration of the NGF antagonist), or according to an independent dosing schedule. In certain embodiments, the NGF antagonist is the only therapeutic agent administered for the treatment of itching or a disease or condition associated with itching (such as atopic dermatitis). For example, in certain embodiments, the subject is not also receiving steroid (such as corticosteroid) treatment. Thus, for example, in some embodiments the subject is not concurrently administered (e.g., not simultaneously or substantially sequentially administered) a steroid, such as a corticosteroid, such as prednisone or prednisolone, or methylprednisolone. In such embodiments, the subject may have been previously treated with a steroid, such as a corticosteroid, but the window of therapeutic effect of the steroid has passed.

The following examples are provided for the purpose of illustration and are not intended to be construed as being limiting on the present invention. Examples

Example 1 - Compositions

Anti-NGF antibody maybe formulated in a phosphate buffered saline composition prepared with 140 mM sodium chloride and 10 mM phosphate, at a pH of 7.4, with anti- NGF antibody added at a concentration of 2 mg/mL.

Anti-NGF antibody may be formulated in a phosphate buffered saline composition with sorbitol prepared as follows:

To 1900 ml of water are added 0.0078 moles (2.15 g) sodium phopshpate monobasic (10 mM), 0.0121 mol (3.41 g) sodium phosphate dibasic (10mM), and 100 g sorbitol. The pH is adjusted to from about 6.7 to about 7.0 with 10N sodium hydroxide and the volume is brought to 2L with addition of water. Anti-NGF antibody is added at a concentration of 2 mg/mL.

Example 2 - Clinical Trial

A dog with atopic dermatitis exhibiting chronic scratching behaviour was administered a single dose of caninized anti-NGF antibody (NV-01) at a dose amount of about 200 μg/kg. (72 mg, supplied at 2 mg/ml). After treatment, the owner observed that the dog was no longer scratching.

Example 3 - Canine Atopic Dermatitis Pilot Study

In atopic dermatitis, increased levels of IL-4 and IL-13 are major pathogenic mediators and have been associated with an increase eosinophil infiltration and NGF production. In inflamed skin lesions, NGF is produced by mast cells, lymphocytes, monocytes, and keratinocytes. Further, expression of both NGF and TrkA are up-regulated in atopic dermatitis lesions, and such expression has been correlated with disease severity or itch. See, e.g., Mollanazar, "Mediators of Chronic Pruritus in Atopic Dermatitis: Getting the Itch Out?" Clin Rev Allergy Immunol. Epub (May 1 , 2015).

Canine atopic dermatitis (CAD) is a multifaceted, cutaneous, chronic disease resulting from a complex interaction between environmental and genetic factors. CAD affects approximately 10% of pure and mixed breed dogs in industrialized countries such as the

United States. Current general recommendations for the treatment of CAD involve a combination of measures including allergen avoidance, the use of anti-inflammatory agents, allergen-specific immunotherapy and antimicrobial drugs.

CAD is often difficult to clinically distinguish from the most common dog skin disorder, flea allergy dermatitis, and other skin diseases such as food allergies, scabies, and demodectic mange. In 2001 , The American College of Veterinary Dermatology Task Force on Canine Atopic Dermatitis proposed the following definition of CAD: "A genetically-predisposed inflammatory and pruritic allergic skin disease with

characteristic clinical features. It is associated most commonly with IgE antibodies to environmental allergens." See, e.g., Olivry et al., Vet. Immunol. Immunopathol. 81 (3- 4): 143-146 (2001). The renamed International Task Force on Canine Atopic Dermatitis (ITFCAD) was established with the aim of increasing standards in CAD diagnosis, and published the third version of the standard tool used for evaluating the severity of lesions associated with CAD, the Canine Atopic Dermatitis Extent and Severity Index (CADESI-03) in 2007. See, e.g., Olivry et al., Vet. Dermatol. 18(2):79-96 (2007).

Subsequently, Plant et al., Vet. Dermatol. 23:515-521 (2012), developed and validated the Canine Atopic Dermatitis Lesion Index (CADLI), a simplified and more efficient tool for evaluating the severity of lesions associated with CAD. The ITFCAD supports adoption of Favrot's criteria for CAD for increasing the homogeneity of enrollment of canine subjects in research trials. See, e.g., Olivry et al. Vet. Dermatol. 21 (1): 124-127 (2010).

NV-01 is a fully caninized anti-NGF monoclonal antibody (mAb) that is under development for the control of pain associated with osteoarthritis (OA) in dogs. NV-01 has a light chain comprising SEQ ID NO:10 of WO 2012/153121 and a heavy chain comprising SEQ ID NO:1 1 of WO 2012/153121. The following describes a pilot study in client-owned dogs evaluating the efficacy of NV- 01 for control of pruritus and resolution of skin lesions in CAD. The study was a multi- center, placebo-controlled, randomized, masked (blinded), pilot study comparing NV-01 with placebo for the control of pruritus and resolution of skin lesions in dogs with CAD.

Client-owned dogs diagnosed with CAD according to Favrot's criteria were enrolled under an established Veterinary Client Patient Relationship (VCPR). A total of 29 evaluable dogs were enrolled, 20 treated with NV-01 and 9 with placebo (saline). The primary efficacy variables were the puritis visual analog score (PVAS) at any time post-treatment compared with the PVAS at screening (e.g. prior to or on Day 0). The CADLI post- treatment at Day 28 and 42 compared to the CADLI at screening also was assessed. As noted above, the PVAS reflects the owner's assessment of itching, while the CADLI represents a veterinarian's assessment of dermatitis based on severity of lesions. With regard to PVAS, success may be defined as improvement in PVAS score of about 2 units or more (corresponding to a difference of about 2 cm or more on the 10 cm scoring scale) compared to PVAS at initial screening, observed at more than one assessment. Pre-Treatment Screening

At a screening visit between Day -7 and Day 0, dogs underwent a physical examination to ensure that they were in good health and appropriate for the study. At the screening visit, a blood sample and a urine sample were collected from each enrolled dog for baseline hematology, serum chemistry and urinalysis. An additional blood sample was collected and stored frozen for possible immunological analysis. The dogs also were evaluated at screening by PVAS and CADLI. Dogs with a CALDI > 5 were eligible for enrolment.

Treatment

On Day 0, dogs received a single subcutaneous injection of NV-01 or placebo in the loose skin over the shoulders or neck. The NV-01 was provided in single use glass vials as a sterile solution containing 2 mg/mL NV-01 in buffered sodium phosphate.

Treatment was administered at a dose of about 0.2 mg/kg of NV-01 per dog or equivalent dose volume of saline (about 0.1 mL/kg). Unit dosing was used, with each dog receiving whole vials (1 mL) according to the dosing schedule shown in Table 1. Partial vials were only used for dogs weighing≤ 5 kg (11.0 lb). TABLE 1

Post-Treatment Evaluation

Dogs were evaluated by PVAS on Day 7 (± 1 ), Day 14 (± 2), Day 21 (± 2), Day 28 (± 3), and Day 42 (± 3). Owners were instructed not to review previous PVAS scores prior to making each assessment. In addition, dogs underwent a comprehensive physical examination and were evaluated by CALDI on Day 28 (± 3) and Day 42 (± 3). Blood samples taken at Day 28 (± 3) were subject to serum chemistry and hematology analysis.

The following were excluded from the effectiveness analysis:

• Dogs withdrawn early from the study due to situations unrelated to the

treatment, such as owner's decision to withdraw the dog from the study or owner non-compliance.

• Dogs withdrawn early due to uncooperativeness, serious injury, or serious illness.

As noted above, an outcome of success may be based on a comparison of the PVAS before and after treatment (e.g., at screening versus Day 7, Day 14, Day 21 , Day 28 or Day 42), and may be defined as improvement in PVAS score of about 2 units or more (about 2 cm or more on the 10 cm scoring scale) compared to PVAS at initial screening. Observation of such an improvement at more than one assessment indicates a meaningful and persistant effect.

The raw PVAS results are set forth in Table 2 below and the results presented relative to PVAS at initial screening is shown in Figures 2 and 3. Seven dogs exhibited an improvement in PVAS score deemed to be meaningful and persistant: one dog in the placebo group (FIG. 2; dog 03-12) and six dogs in the NV-01 group (FIG. 4, dogs 03-04, 03-11 , 03-09, 03-05,03-07 and 03-18. Thus, the NV-01 group had a higer rate of success following treatment than the placebo group.

CALDI scores did not show a significant difference between the placebo group and NV-01 treated group during the assessment period, but long term benefits are expected.

Table 2

Dog Dav O Dav 7 Dav 14 Dav 21 Dav 28 Dav 42

Placebo

01-01 7.0 7.5 7.1 7.3 7.3 7.6

01-05 7.1 6.9 6.0 6.4 5.7 8.1

02-03 9.2 9.1 8.7 8.7 9.1 9.2

02-04 7.1 5.8 6.4 7.2 7.0 7.3

03-01 5.7 5.8 5.8 5.4 5.9 5.6

03-08 6.2 7.9 7.4 6.6 6.8 5.4

03-12 7.5 7.3 7.0 3.8 2.3 2.7

03-15 7.3 5.0 7.1 7.1 7.0 6.9

03-17 6.1 5.8 5.7 6.2 6.7 6.1

NV-01

01-03 5.5 7.3 — — — —

01-04 6.4 6.6 8.0 8.0 8.1 7.6

01-08 7.2 7.0 7.3 6.9 7.7 6.9

02-01 7.8 8.1 8.1 8.4 8.2 7.2

02-05 6.7 6.4 6.4 6.5 5.9 6.5

03-03 5.2 4.9 6.7 7.0 7.1 7.1

03-04 7.1 1.8 0.7 1.0 0.2 1.6 Dog Dav O Dav 7 Dav 14 Dav 21 Dav 28 Dav 42

03-09 7.0 7.1 7.1 3.0 3.1 3.1

03-11 9.2 8.7 6.7 3.8 2.9 2.3

03-14 5.4 7.5 9.7 9.7 9.8 9.6

03-18 8.1 7.3 6.5 6.0 5.3 6.6

03-19 6.0 6.0 — 6.2 6.2 6.1

01-02 6.9 6.3 5.2 6.4 — —

02-02 7.2 7.6 7.4 7.6 5.5 5.6

03-02 7.2 6.9 7.2 7.9 7.7 8.1

03-05 5.0 5.2 3.8 2.9 2.5 2.5

03-07 7.1 7.2 5.1 5.1 5.2 5.2

03-10 5.2 — 6.6 8.1 7.0 7.6

03-13 7.3 6.7 5.5 7.5 7.2 6.6

03-16 7.2 7.6 7.6 7.1 7.6 7.2

All documents referred to in this specification are herein incorporated by reference. Various modifications and variations to the described embodiments of the inventions will be apparent to those skilled in the art without departing from the scope of the invention. Although the invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes of carrying out the invention which are obvious to those skilled in the art are intended to be covered by the present invention. between the placebo group and NV-01 treated group

Claims

1. An NGF antagonist for use in a method of treating itching in a subject in need thereof.

2. The NGF antagonist for use according to claim 1 , wherein the subject is human, equine, canine or feline.

3. The NGF antagonist for use according to claim 1 , wherein the itching is chronic itching.

4. The NGF antagonist for use according to claim 1 , wherein the itching is acute itching.

5. The NGF antagonist for use according to claim 1 , wherein the itching is associated with one or more conditions selected from the group consisting of skin diseases, skin conditions, contact skin reactions, cancer, kidney disease, liver disease, viral infections, parasites, food allergies, and drug reactions.

6. The NGF antagonist for use according to claim 1 , wherein the itching is associated with atopic dermatitis.

7. The NGF antagonist for use according to claim 1 , wherein the itching is idiopathic itching.

8. The NGF antagonist for use according to any one of claims 1-7, wherein the NGF antagonist is an anti-NGF antibody or NGF-binding antibody fragment.

9. The NGF antagonist for use according to claim 8, wherein the subject is human and the NGF antagonist is a humanized anti-NGF antibody or humanized NGF-binding antibody fragment.

10. The NGF antagonist for use according to claim 9, wherein the subject is human and the NGF antagonist is tanezumab.

11. The NGF antagonist for use according to claim 8, wherein the subject is equine and the NGF antagonist Is an equinized and-NGF antibody or equinized NGF-binding antibody fragment

12. The NGF antagonist for use according to claim 8, wherein the subject Is canine and the NGF antagonist is a caninized anti-NGF antibody or caninized NGF-binding antibody fragment

13. The NGF antagonist for use according to claim 8, wherein the subject is feline and the NGF antagonist is a felinized anti-NGF antibody or felinized NGF-binding antibody fragment

14. The NGF antagonist for use according to any one of claims 1-7, wherein the NGF antagonist is an NGF-binding polypeptide.

15. The NGF antagonist for use according to claim 1 , wherein the NGF antagonist is selected from the group consisting of polypeptides comprising TrkA and polypeptides comprising p75, wherein the polypepUdes optionally further comprise an immunoglobulin Fc region.

16. The NGF antagonist for use according to claim 1. wherein the NGF antagonist is administered in a pharmaceutical composition.

1 . The NGF antagonist for use according to claim 1. wherein the NGF antagonist is administered parenterally.

18. The NGF antagonist for use according to claim 1 , wherein the NGF antagonist is administered intravenously orsubcutaneously.

19. The NGF antagonist for use according to claim 1. wherein the subject is suffering from chronic itching and the NGF antagonist is administered once every four to six weeks.

20. The NGF antagonist for use according to any one of claims 1-7, wherein the use further comprises administering an additional agent useful against itching.

2 . The NGF antagonist for use according to any one of claims 1-7, wherein the subject is not receiving concurrent steroid treatment

22- Use of an NGF antagonist in the manufacture of a medicament for use in a method of treating itching in a subject in need thereof.

23. The use according to claim 22, wherein the subject is human, equfne. canine or feline.

24. The use according to claim 22, wherein the itching is chronic itching, acute itching, and or is associated with one or more conditions selected from the group consisting of skin diseases such as atopic dermatitis, skin conditions, contact skin reactions, cancer, kidney disease, liver disease, viral infections, parasites, food allergies, drug reactions, or idiopathic itching.

25. The use according to claim 22, wherein the NGF antagonist is an anti-NGF antibody or NGF-binding antibody fragment

26. The use according to claim 22, wherein (i) the subject is human and the NGF antagonist is a humanized anti-NGF antibody or humanized NGF-binding antibody fragment or is tanezumab; (ii) wherein the subject is equine and the NGF antagonist is an equinized anti-NGF antibody or equinized NGF-binding antibody fragment; (lli) the subject is canine and the NGF antagonist is a caninized anti-NGF antibody or caninized NGF-binding antibody fragment; or (iv) the subject is feline and the NGF antagonist is a felinized anti-NGF antibody or felinized NGF-binding antibody fragment

27. The use according to claim 22, wherein the NGF antagonist is an NGF-binding polypeptide, such as polypeptides comprising TrkA and polypeptides comprising p75, optionally further comprising an immunoglobulin Fc region.

28. The use according to claim 22, wherein the subject is suffering from chronic itching and the NGF antagonist is administered pareneterally once every four to six weeks.

29. The use according to claim 22, wherein the method further comprises

administering to the subject an additional agent useful against itching.

30. The use according to claim 22, wherein in accordance with the method the subject is not receiving concurrent steroid treatment.

31. A method of treating itching in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an NGF antagonist.

32. The method of claim 31 , wherein the subject is human, equine, canine or feline.

33. The method of claim 31 , wherein the itching is chronic itching.

34. The method of claim 31 , wherein the itching is acute itching.

35. The method of claim 31 , wherein the itching is associated with one or more conditions selected from the group consisting of skin diseases, skin conditions, contact skin reactions, cancer, kidney disease, liver disease, viral infections, parasites, food allergies, and drug reactions.

36. The method of claim 31 , wherein the itching is associated with atopic dermatitis.

37. The method of claim 31 , wherein the itching is idiopathic itching.

38. The method of claim 31 , wherein the NGF antagonist is an anti-NGF antibody or NGF-binding antibody fragment.

39. The method of claim 38, wherein the subject is human and the NGF antagonist is a humanized anti-NGF antibody or humanized NGF-binding antibody fragment.

40. The method of claim 39, wherein the subject is human and the NGF antagonist is tanezumab.

41. The method of claim 38, wherein the subject is equine and the NGF antagonist is an equinized anti-NGF antibody or equlnized NGF-binding antibody fragment.

42. The method of claim 38, wherein the subject is canine and the NGF antagonist Is a carn^'nized anti-NGF antibody orcaninized NGF-binding antibody fragment

43. The method of claim 38, wherein the subject is feline and the NGF antagonist is a felinized anti-NGF antibody or felinized NGF-binding antibody fragment

44. The method of claim 31 , wherein the NGF antagonist is an NGF-binding polypeptide.

45. The method of claim 44, wherein the NGF antagonist is selected from the group consisting of polypeptides comprising TrkA and polypeptides comprising p75, wherein the polypeptides optionally further comprise an immunoglobulin Fc region.

46. The method of claim 31 , wherein the NGF antagonist is administered in a pharmaceutical composition,

47. The method of claim 31 , wherein the NGF antagonist is administered parenteraliy.

48. The method of claim 31 , wherein the NGF antagonist is administered intravenously or subcutaneously.

49. The method of claim 31 , wherein the subject is suffering from chronic itching and the NGF antagonist is administered once every four to six weeks.

50. The method of claim 31, further comprising administering an additional agent useful against itching.

51. The method of claim 31 , where the subject is not receiving concurrent steroid treatment.

52. An NGF antagonist, for use in a method of treating itching in a subject in need thereof, as set forth in any one of claims 31-51.

53. Use of an NGF antagonist in the manufacture of a medicament for use in a method of treating itching in a subject in need thereof, as set forth in any one of claims 31-51.