WO2006108128A2

WO2006108128A2 - Ocular delivery of interferon-tau

Info

Publication number: WO2006108128A2
Application number: PCT/US2006/012980
Authority: WO
Inventors: Chih-Ping Liu
Original assignee: Pepgen Corporation
Priority date: 2005-04-05
Filing date: 2006-04-05
Publication date: 2006-10-12
Also published as: WO2006108128A3; JP2008534690A; AU2006232130A1; CA2602850A1

Abstract

Methods for treating ocular disorders and systemic diseases by administering interferon-tau to the eye are described.

Description

OCULAR DELIVERY OF INTERFERON-TAU

TECHNICAL FIELD

The subject matter described herein relates to ocular delivery of interferon- tau for treatment of eye disorders or systemic diseases.

BACKGROUND

Opthalmic preparations, including solutions, suspensions, ointments, and ocular inserts, provide a convenient route of administering therapeutic drugs to human patients. Drugs can be applied topically to the eye to exert a local effect in the eye or a systemic effect after absorption into the bloodstream via, for example, the blood vessels in the conjunctival mucosa.

Interferon-tau has been administered orally for treatment of systemic autoimmune conditions and viral infections (Soos, J. M. etal., J. Neuroimmunology, 75:43-50 (1997); Soos, J.M. et al., J. Immunology, 169(5): 2231-2235 (2002);

Nakajima, A. etal., J. Interferon Cytokine Res., 22:397-401 (2002)). There has been no suggestion in the art, however, of administering the protein to the eye for treatment of systemic disorders or for treatment of local disorders in the eye.

The foregoing examples of the related art and limitations related therewith are intended to be illustrative and not exclusive. Other limitations of the related art will become apparent to those of skill in the art upon a reading of the specification and a study of the drawings.

BRIEF SUMMARY The following aspects and embodiments thereof described and illustrated below are meant to be exemplary and illustrative, not limiting in scope.

In one aspect, a treatment method comprising administering interferon-tau to the eye of a patient is described. In one embodiment, the patient is suffering from a disorder, systemic or localized ocular, that is responsive to interferon-tau. In another embodiment, interferon-tau is administered to the eye in the form of a solution, a suspension, or an ointment. These preparations can be formulated, in other embodiments, for sustained release. Ocular devices are also contemplated, such as an ocular insert. In other embodiments, the interferon-tau has a certain degree of identity to the protein identified herein as SEQ ID NO:2.

In another embodiment, the interferon-tau is formulated with histidine, for increased stability and/or solubility in aqueous formulations. In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to the drawings and by study of the following descriptions.

BRIEF DESCRIPTION OF THE SEQUENCES SEQ ID NO:1 corresponds to an amino acid sequence of mature ovine interferon-τ (IFNτ; oTP-1 ; GenBank Accession No. Y00287; PID g1358).

SEQ ID NO:2 corresponds to an amino acid sequence of mature ovine IFNx, where the amino acid residues at positions 5 and 6 of the sequence are modified relative to the sequence of SEQ ID NO:1.

DETAILED DESCRIPTION I. Definitions

Interferon-tau, abbreviated as IFNτ or interferon-τ, refers to any one of a family of interferon proteins having at least one characteristic from each of the following two groups of characteristics: (i) (a) anti-luteolytic properties, (b) antiviral properties, (c) anti-cellular proliferation properties; and (ii) about 45 to 68% amino acid homology with α-interferons and greater than 70% amino acid homology to known IFNτ sequences (e.g., Ott, et al., J. Interferon Res., VV.357 (1991 ); Helmer, et al., J. Reprod. Fert., 79:83 (1987); Imakawa, et al., MoI. Endocrinol, 3:127 (1989); Whaley, et al., J. Biol. Chem., 269:10846 (1994); Bazer, et al., WO 94/10313 (1994)). Amino acid homology can be determined using, for example, the LALIGN program with default parameters. This program is found in the FASTA version 1.7 suite of sequence comparison programs (Pearson and Lipman, PNAS, 85:2444 (1988); Pearson, Methods in Enzymology, 183:63 (1990); program available from William R. Pearson, Department of Biological Chemistry, Box 440, Jordan Hall, Charlottesville, VA). IFNτ sequences have been identified in various ruminant species, including but not limited to, cow (Bovine sp., HelmerS.D., J. Reprod. Fert., 79:83 (1987); Imakawa, K., MoI. Endocrinol., 119:532 (1988)), sheep (Ovine sp.), musk ox (Ovibos sp.), giraffe (Giraffa sp., GenBank Accession no. U55050), horse {Equus caballus), zebra {Eqυus burchelli, GenBank Accession no. NC005027), hippopotamus (Hippopotamus sp.), elephant (Loxodonta sp.), llama (Llama glama), goat (Capra sp., GenBank Accession nos. AY357336, AY357335, AY347334, AY357333, AY357332, AY357331 , AY357330, AY357329, AY357328, AY357327), and deer (Cervidae sp.). The nucleotide sequences of IFNτ for many of these species are reported in public databases and/or in the literature (see, for example, Roberts, R.M. et ai, J. Interferon and Cytokine Res., 18:805 (1998), Leaman D.W. et al., J. Interferon Res., 12:1 (1993), Ryan, A.M. et ai, Anim. Genet, 34:9 (1996)). The term "interferon-tau" intends to encompass the interferon-tau protein from any ruminant species, exemplified by those recited above, that has at least one characteristic from each of the following two groups of characteristics listed above.

Ovine IFNτ (OvlFNτ) refers to a protein having the amino acid sequence as identified herein as SEQ ID NO:1, and to proteins having amino acid substitutions and alterations such as neutral amino acid substitutions that do not significantly affect the activity of the protein, such as the IFNτ protein identified herein as SEQ ID NO:2. More generally, an ovine IFNτ protein is one having about 80%, more preferably 90%, sequence homology to the sequence identified as SEQ ID NO:1. Sequence homology is determined, for example, by a strict amino acid comparison or using one of the many programs commercially available.

Treating a condition refers to administering a therapeutic substance effective to reduce the symptoms of the condition and/or lessen the severity of the condition. A condition "responsive to interferon therapy" is one in which the existence, progression, or symptoms of the condition is altered upon administration of an interferon, in particular a type-l interferon, and more particularly, interferon-tau. More preferably, a condition responsive to interferon therapy is one where the existence, progression, or symptoms of the condition are alleviated by IFNτ administered parenterally or orally. II. Treatment Methods

In one aspect, a treatment method comprising administering IFNτ to the eye of a human patient is provided. As will be described below, the method finds use in treating localized eye disorders and in treating systemic disorders, that are responsive to IFNτ. IFNx is a type I IFN first identified as a pregnancy recognition hormone in ruminants, such as sheep and cows (Bazer, F. VV. et al., Am. J. Reprod. Immunol. 26:19-22 (1991)). The protein possesses antiviral and anti-proliferative properties, with considerably lower toxicity than other type I interferons (Pontzer, C₁ et al., Biochem. Biophys. Res. Comrn., 152(2):801-807 (1988); Pontzer, C₁ et al., Cancer Res., 51;5304 (1991 )). Relative to other interferons, ovine IFNτ shares about 45-55% identity with IFN-αs from human, mouse, rat, and pig and 70% homology with bovine IFN-αll, now referred to as IFN-Ω. A cDNA of ovine IFNτ and several cDNA sequences which may represent different isoforms have been reported in the literature (Imakawa, K. et al, Nature, 330:377-379, (1987); Stewart, H.J., et al, . MoI. Endocrinol. 2:65 (1989); Klemann, S. W., et al., Nuc. Acids Res. 18:6724 (1990); and Charlier, M., et al., MoI. Cell Endocrinol. 76:161-171 (1991)). All are approximately 1kb with a 585 base open reading frame that codes for a 23 amino acid leader sequence and a 172 amino acid mature protein.

The 172 amino acid sequence of ovine-IFNτ is set forth, for example, in U.S. Patent No. 5,958,402, and its homologous bovine-IFNτ sequence is described, for example, in Helmer et al., J. Reprod. Fert., 79:83-91 (1987) and Imakawa, K. et al., MoI. Endocrinol., 3:127 (1989). The sequences of ovine-IFNτ and bovine-IFNτ from these references are hereby incorporated by reference. An amino acid sequence of ovine IFNτ is shown herein as SEQ ID NO:1. A modified amino acid sequence of ovine IFNτ is shown herein as SEQ ID NO:2.

Recombinant production of IFNτ is described in both the scientific literature (Ott, et al., J. Interferon Cytokine Res., 11:357-364 (1991); Soos, J. M. et al., J. Immunol., 155:2747 (1995)) and the patent literature (WO/94/10313; US 2003/0049277, the description of IFNτ production in these documents is incorporated by reference herein.)

IFNτ is formulated in any pharmaceutically-acceptable vehicle for delivery to the eye. In one embodiment, the formulation is a solution or suspension or ointment that comprises lFNτ. The formulation may contain any physiologically compatible vehicle, as those skilled in the ophthalmic art can select using conventional criteria. The vehicles may be selected from the known ophthalmic vehicles which include, but are not limited to, saline solution, water, mineral oil, petroleum jelly, polyethers such as polyethylene glycol, polyvinyls such as polyvinyl alcohol and povidone, cellulose derivatives such as methylcellulose and hydroxypropyl methylcellulose, petroleum derivatives such as mineral oil and white petrolatum, animal fats such as lanolin, vegetable oils such as olive oil, polymers of acrylic acid such as carboxypolymethylene gel, vegetable fats such as peanut oil and polysaccharides such as dextrans, and glycosaminoglycans such as sodium hyaluronate, salts such as sodium chloride and potassium chloride, alcohols such as ethanol, n-propyl alcohol, and iso-propyl alcohol, and dimethyl sulfoxide. These vehicles can be used alone in various combinations.

The formulation can also be designed for sustained or controlled release in the eye. For example, formulations where the protein is entrapped or incorporated into particles, such as lipidic particles or polymeric particles, are contemplated.

The INFτ formulation is administered topically as an ophthalmic drop (solution or suspension) or ophthalmic ointment containing an amount of the protein. Typically, the preparation is administered in any quantity needed to provide the degree of treatment needed. For example, drops can be instilled into the eye multiple times throughout the day, for example, hourly, every 2-3 hours, or several times per day.

Delivery devices designed for ocular administration of drugs are also suitable for use in the treatment method. Exemplary devices include Ocusert^® and Lacrisert^®. In these devices, a reservoir of the active agent is prepared, the reservoir being a matrix, gel, or fluid. The reservoir can be surrounded by additional polymer membrane layers, such as the ethylene-vinyl acetate membranes in the Ocusert^® device. The device is placed in the conjunctival sac where the drug is released for a prolonged period of time.

The preparations described above can additionally include a preservative, a surfactant, a viscosity enhancer, a buffer. In one preferred embodiment, the preparation contains with histidine, which acts to stabilize the tertiary structure of (FNx and to increase its solubility in aqueous formulations, as described in WO 2004/032863, which is incorporated by reference herein. In one embodiment, an ocular preparation as described above is administered for treatment of uveitis. Uveitis is an inflammation of the uvea and is responsible for about 10% of the visual impairment in the U.S. Panuveitis refers to inflammationof the entire uveal (vascular) layer of the eye. Posterior uveitis generally refers to chorioentinitis and anterior uveitis refers to iridocyclitis. The inflammatory precuts, cells, fibrins, excess proteins, of these inflammations are commonly found in the fluid spaces of the eye, i.e., the anterior changer, posterior chamber, and vitreous space as well as infiltrating the tissue intimately involved in the inflammatory response. Uveitis may occur following surgical or traumatic injury to the eye or as a component of another disorder, such as an autoimmune disease. Accordingly, the invention contemplates administering to a person suffering from uveitis an amount of IFNτ delivered to the eye for treatment of the condition or relief of the symptoms. In another embodiment, an ocular preparation as described above is administered for treatment of Sjogren's syndrome. Sjogren's syndrome is an autoimmune disease in which the body's immune system mistakenly attacks its own moisture producing glands. The syndrome can occur alone or in combination with another connective tissue disorder, such as rheumatoid arthritis, systemic lupus, systemic sclerosis (scleroderma) or polymyositis/dermatomyositis. Hallmark symptoms of Sjogren's syndrome are dry eyes and dry mouth, but it is a systemic disease, affecting many organs. Accordingly, it is contemplated to administer to a person suffering from Sjogren's syndrome an amount of IFNτ delivered to the eye for treatment of the condition or relief of the symptoms, both locally in the eye and systemically.

In another embodiment, an ocular preparation as described above is administered for treatment of a person at risk of an ocular tissue transplantation. Corneal transplant, also known as keratoplasty, involves replacing a patient's damaged cornea with a cornea from the eye of a human cadaver. Corneal transplant is done when vision is lost in an eye because the cornea has been damaged by disease or traumatic injury. Some of the disease conditions that might require corneal transplant include the bulging outward of the cornea

(keratoconus), a malfunction of the inner layer of the cornea (Fuchs' dystrophy), and painful swelling of the cornea (pseudophakic bullous keratopathy). Some of these conditions cause cloudiness of the cornea; others alter its natural curvature, which can also reduce the quality of vision. Injury to the cornea can occur because of chemical burns, mechanical trauma, or infection by viruses, bacteria, fungi, or protozoa. Herpes virus is one of the more common infections leading to corneal transplant. Accordingly, it is contemplated to administer to a person having a cornea transplant an amount of IFNτ delivered to the eye for prevention of tissue rejection. More generally, the invention contemplates administering IFNτ to the eye in an amount to inhibit rejection of a tissue or organ transplant.

In another embodiment, an ocular preparation as described above is administered for treatment of a systemic disorder that is responsive to IFNτ. Systemic disorders include autoimmune, inflammatory, viral infections, proliferative and hyperproliferative diseases, as well as immunologically-mediated diseases. An ophthalmic preparation comprising INFτ is prepared and delivered in an amount sufficient to treat the condition or ameliorate the symptoms associated with the condition.

It will be appreciated that the ocular iFNτ preparation can be administered alone or in combination with another therapeutic agent, given by any route of administration. Selection of a second agent is made by a primary caregiver and typically relates to the condition from which the patient suffers. For example, as noted above, Sjogren's syndrome is often accompanied by another condition, such as rheumatoid arthritis. IFNτ is administered to the eye for local treatment of Sjogren's syndrome and a second agent, such as a pain reliever or a steroid, is administered parenterally or orally for treatment of the conditions associated with rheumatoid arthritis. By way of another example, IFNτ can be administered to the eye for treatment of a systemic autoimmune condition, such as multiple sclerosis, along with a second drug, such as azathioprine, cyclophosphamide, corticosteroids (prednisone, prednisolone, others), cyclosporine, mycophenolate mofetil, antithymocyte globulin, muromonab-CD3 monoclonal antibody, mercaptopurine, mitoxantrone, glatiramer acetate (Copaxone), interferon-beta (Avonex™, Betaseron™, Ribif™), daclizumab, methotrexate, sirolimus, tacrolimus, and others. The second drug can be administered orally, parenterally, or topically. Selection of other second therapeutic agents for combination with IFNτ for a variety of conditions are readily identified by trained medical personnel. III. Examples

The following examples are illustrative in nature and are in no way intended to be limiting.

EXAMPLE 1

In vivo Ocular Delivery to Mice

Eight Balb/c-mice are obtained for use in a study to evaluate the local and systemic effects of interferon-tau administered to the eye. The mice are housed in standard cages in one room under controlled environmental conditions. Animals have free access to food and water throughout the study. Animals are observed daily for signs of ill health and only healthy animals with no ocular abnormalities are used for experiments.

Interferon-tau (SEQ ID NO:2) is prepared according to well known techniques and the specific activity is measured in a standard cytopathic effect assay (see, e.g., US 2005-0142109). The protein is formulated in water and NaCI to make an isotonic solution. The solution is instilled three times daily into the conjunctival sac of the right eye of each mouse in drop form (10-100 μL) to provide a dose of between 0.001-10 μg of interferon-tau.

After treatment for various periods of time, mice are sacrificed. Blood and tissue samples are collected for analysis of immunological markers, including IL-10, IL-12, IFN-gamma, other pro-inflammatory cytokines, chemokines, cellular markers, and biological markers, such as 2'-5'-oligoadenylate synthetase (OAS), indolamine dioxygenase, and interferon response genes, such as type I receptors, interferon- response factor 1 , MxA protein, neopterine, JAK, stat proteins.

EXAMPLE 2

In vivo Ocular Delivery to Rabbits

For a study to evaluate the acute ocular safety of interferon-tau, five healthy, male, adult albino New Zealand rabbits (2-2.5 kg) are obtained. Animals are housed in standard cages in one room under controlled environmental conditions. Animals have free access to food and water throughout the study. Animals are observed daily for signs of ill health and only healthy animals with no ocular abnormalities are used for experiments. Interferon-tau (SEQ ID NO:2) is formulated in water and NaCI to make an isotonic solution. The solution is instilled into the conjunctival sac of the right eye of each rabbits by multiple 50 μL instillations (5 times in 20 minutes). Animals are assigned a clinical ocular safety grade for the conjunctiva, cornea and iris according to a modified Draize scale at 0, 1 , 2 and 3 hours after the last instillation. The left eye is instilled with physiological saline and serves as a control.

EXAMPLE 3

In vivo Ocular Delivery to Patient Suffering from Uveitis Patients diagnosed with uveitis are enrolled for treatment with interferon-tau.

Interferon-tau is formulated in saline and histidine buffer to instillation into both eyes. One drop of approximately 40 microliters is administered to the patient every two hours during waking hours for five consecutive days.

The acute phase and flare-ups of uveitis are measured using methods well known in the art, including symptoms such as increased clouding of the cornea, appearance of precipitates and new synechiae. Clinical signs of the remission of uveitis are measured using clinical parameters such as disappearance of corneal clouding, absence of vasodilation in the iris. A diminished amount of precipitates and synechiae indicate remission.

EXAMPLE 4

In vivo Ocular Delivery to Patient Suffering from Sjogren's Syndrome Patients diagnosed with Sjogren's syndrome are enrolled for a study to evaluate the use of interferon-tau for treatment of salivary hypofunction and dry mouth symptoms in primary Sjogren's syndrome patients. Interferon-tau is formulated in saline and histidine buffer to instillation into both eyes. One drop of approximately 40 microliters is administered to the patient every two hours during waking hours for twenty weeks. A separate group of patients are treated with a placebo formulation. Effectiveness of interferon-tau is evaluated using a Dry Eye Questionnaire that includes categorical scales to measure the prevalence, frequency, diurnal intensity, and intrusiveness of common ocular surface symptoms, including discomfort, dryness, visual changes, soreness and irritation, grittiness and scratchiness, burning and stinging, foreign body sensation, light sensitivity, and itching. During the treatment period, unstimulated whole saliva flow, stimulated whole saliva flow, and oral dryness are measured periodically in each treated and placebo patient. In addition, a Schirmer test for tear production is used to evaluate increase in tear production in treated patients.

While a number of exemplary aspects and embodiments have been discussed above, those of skill in the art will recognize certain modifications, permutations, additions and sub-combinations thereof. It is therefore intended that the following appended claims and claims hereafter introduced are interpreted to include all such modifications, permutations, additions and sub-combinations as are within their true spirit and scope.

Claims

IT IS CLAIMED:

1. Use of interferon-tau for the manufacture of a medicament for treating a disease responsive to treatment by interferon-tau by administering said medicament to the eye of a subject.

2. The use according to claim 1 , wherein said medicament is in the form of a solution, a suspension, or an ointment.

3. The use according to claim 1 , wherein said medicament is in the form of an ocular insert.

4. The use according to claim 1 , wherein said medicament is in the form of a sustained release dosage form.

5. The use according to claim 1 , wherein said interferon-tau has a sequence at least 80% identity to SEQ ID NO:2.

6. The use according to claim 1 , wherein said interferon-tau has a sequence identified as SEQ ID NO:1 or SEQ ID NO:2.

7. The use according to claim 1 , wherein said medicament further comprises histidine.

8. The use according to claim 1 , wherein said subject is suffering from a disorder to an eye selected from uveitis and Sjogren's syndrome.

9. The use according to claim 1 , wherein said subject is suffering from a systemic disorder.

10. The use according to claim 1 , wherein said subject is treated with a second therapeutic agent.

1 1. A treatment method, comprising administering to the eye an effective amount of interferon-tau.

12. The method of claim 1 , wherein said administering comprises administering interferon-tau in the form of a solution, a suspension, or an ointment.

13. The method of claim 1 , wherein said administering comprises administering interferon-tau in the form of an ocular insert.

14. The method of claim 1 , wherein said administering comprises administering interferon-tau in a sustained release dosage form.

15. The method of claim 1 , wherein said administering comprises administering an interferon-tau having a sequence at least 80% identity to SEQ ID NO:2.

16. The method of claim 1 , wherein said administering comprises administering an interferon-tau having a sequence identified herein as SEQ ID NO:1 or SEQ ID NO:2.

17. The method of claim 1 , wherein said administering comprises administering a preparation comprising histidine.

18. The method of claim 1 , wherein said administering comprises administering to a patient suffering from a disorder to an eye.

19. The method of claim 18, wherein said eye disorder is selected from uveitis and Sjogren's syndrome.

20. The method of claim 18, wherein said administering comprising administering to a patient at risk of rejection of an eye tissue transplant.

21. The method of claim 20, wherein said tissue transplant is selected from cornea and lens.

22. The method of claim 1 , wherein said administering comprises administering to a patient suffering from a systemic disorder.

23. The method of claim 1 , further comprising administering a second therapeutic agent to the patient.

24. The method of claim 23, wherein said second agent is administered parenterally or orally.