COMPOSITIONS AND METHODS RELATING TO THE TREATMENT OF DISEASES

Abstract

The present invention relates to compositions and methods for preventing or treating pruritus, prurigo, neutrophilic dermatoses and skin cancer. The invention extends to compositions for preventing or treating conditions where an exaggerated Th2 response plays a detrimental role. The invention further extends to compositions and the use of the compositions of the invention for the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses and skin cancer for Human and Veterinary therapies.

Claims

1. A method for the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer, said method comprising the step of: (i) administering to a subject in need thereof a therapeutically effective amount of an interferon alpha subtype, wherein the interferon alpha subtype is IFN-α14, HYBRID 1, HYBRID 2 or a combination of at least two of IFN-α14, HYBRID 1 and HYBRID 2.

2. The method of claim 1, wherein the interferon alpha subtype IFN-α14 comprises or consists of an amino acid sequence SEQ ID NO: 1 or a functionally active fragment or variant thereof.

3. The method of claim 1, wherein the interferon alpha subtype HYBRID 1 comprises or consists of an amino acid sequence SEQ ID NO:2 or a functionally active fragment or variant thereof.

4. The method of claim 1, wherein the interferon alpha subtype HYBRID 2 comprises or consists of an amino acid sequence SEQ ID NO:3 or a functionally active fragment or variant thereof.

5. The method of claim 1, wherein the method of administration is selected from topical administration and sublingual administration.

6. The method of claim 1, wherein the therapeutically effective amount of the interferon alpha subtype is a low dose.

7. The method of claim 1, wherein the skin cancer is cutaneous T-cell lymphoma.

8. The method of claim 1, for treatment of a condition selected from pruritus, prurigo, or neutrophilic dermatoses.

9. (canceled)

10. The method of claim 8, wherein the neutrophilic dermatoses is at least one selected from the list comprising sweet syndrome (SS), neutrophilic eccrine hidradenitis (NEH), Behcet disease (BD), pyoderma gangrenosum (PG), bowel-associated dermatosis-arthritis syndrome, rheumatoid neutrophilic dermatitis, adult Still disease, primarily bullous, epidermal, and vasculitic NDs.

11. (canceled)

12. An interferon alpha subtype, wherein the interferon alpha subtype is IFN-α14, HYBRID 1, HYBRID 2 or a combination of at least two of IFN-α14, HYBRID 1,_and_HYBRID 2 for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer.

13. The interferon alpha subtype of claim 12 for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer, wherein the IFN-α14 comprises or consists of an amino acid sequence SEQ ID NO:1 or a functionally active fragment or variant thereof.

14. The interferon alpha subtype of claim 12 for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer, wherein the interferon alpha subtype HYBRID 1 comprises or consists of an amino acid sequence SEQ ID NO:2 or a functionally active fragment or variant thereof.

15. The interferon alpha subtype of claim 12 for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer, wherein the interferon alpha subtype HYBRID 2 comprises or consists of an amino acid sequence SEQ ID NO:3 or a functionally active fragment or variant thereof.

16. The interferon alpha subtype of claim 12 for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer, wherein the interferon alpha subtype is administered topically or is by sublingual administration.

17. The interferon subtype of claim 16 for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer, wherein the interferon alpha subtype is administered at a low dose.

18. The interferon alpha subtype of claim 12 for use in the treatment of skin cancer wherein, the skin cancer is cutaneous T-cell lymphoma.

19. The interferon alpha subtype of claim 12 for use in the treatment of a condition selected from pruritus, prurigo, or neutrophilic dermatoses.

20. (canceled)

21. The interferon alpha subtype of claim 19 for use in the treatment of neurophilic dermatoses wherein the neutrophilic dermatoses are selected from the list comprising sweet syndrome (SS), neutrophilic eccrine hidradenitis (NEH), Behcet disease (BD), pyoderma gangrenosum (PG), bowel-associated dermatosis-arthritis syndrome, rheumatoid neutrophilic dermatitis, adult Still disease, primarily bullous, epidermal, and vasculitic NDs.

22. The interferon alpha subtype of claim 19 for use in the treatment of nucleophilic dermatoses wherein the neutrophilic dermatoses are selected from the list comprising sweet syndrome (SS), neutrophilic eccrine hidradenitis (NEH), Behcet disease (BD), pyoderma gangrenosum (PG), bowel-associated dermatosis-arthritis syndrome, rheumatoid neutrophilic dermatitis, and adult Still disease.

23. (canceled)

24. A composition comprising an interferon alpha subtype, wherein the interferon alpha subtype is IFN-α14, HYBRID 1, HYBRID 2 or a combination of IFN-α14 and HYBRID 1 or HYBRID 2, for use in the treatment and/or prophylaxis of pruritus, prurigo, neutrophilic dermatoses or skin cancer.

25. (canceled)

Description

BRIEF DESCRIPTION OF THE FIGURES

[0131] FIG. 1 shows a graph demonstrating the effect of IFNα-14 on CXCL8 (IL8) production from human keratinocytes.

[0132] FIG. 2 shows a graph demonstrating the effect of IFNα-14 on IL6 production from human keratinocytes.

[0133] FIG. 3 shows a graph demonstrating the effect of IFNα-14 on IL31 production from human leucocytes.

[0134] FIG. 4 shows a graph demonstrating the effect of HYBRID 1 on IL31 production from human leucocytes.

[0135] FIG. 5 shows a graph demonstrating the effect of HYBRID 2 on IL31 production from human leucocytes.

[0136] FIG. 6 shows a graph demonstrating the effect of Human Alpha-Interferon-2a (Roferon) on IL31 production from human leucocytes.

[0137] FIG. 7 shows the IFN-α14 amino acid sequence.

[0138] FIG. 8 shows the HYBRID 1 amino acid sequence.

[0139] FIG. 9 shows the HYBRID 2 amino acid sequence.

[0140] FIG. 10 demonstrates the pruritic pathway.

Experimental Data

Experiment 1: The Effect Of IFNα-14 On IL-6 And CXCL8 (IL8) Production In Ketatinocytes From Normal Human Skin

[0141] The inventors tested the effect of IFNα-14 on keratinocytes from normal human skin that were activated with TNF-α to induce chemokine secretion.

[0142] FIG. 1 demonstrates that IFNα-14 suppresses CXC18 (IL8) secretion directly by >80%. FIG. 1 indicates strong inhibition of CXCL8 (IL8) in the presence of IFNα-14 and in particular at low concentrations of IFNα-14. CXCL8 is a major contributor to inflammation. It attracts neutrophils and basophils/mast cells to the site of inflammation - the latter release histamine and many other noxious agents e.g. prostaglandins, leukotrienes.

[0143] FIG. 2 demonstrates inhibition of IL6 production in the presence of IFNα-14 and in particular at low concentrations of IFNα-14. IL-6 is an acute phase reactant that rises in trauma and is widely employed as an ancillary growth factor/stimulant. IL6 is a growth factor commonly associated with stress.

Experiment 2: The Effect Of IFNα-14, HYBRID 1 And HYBRID2 On IL31 Production In Human Leucocytes

Whole Blood Stimulation Assay

[0144] Fresh whole human blood (50 IU/ml preservative heparin) was diluted ⅒ with RPMI 1640 medium with penicillin/streptomycin, L-glutamine and calcium. This was incubated with or without PHA-P (100 .Math.g/ml) in the presence of a range of concentrations of either rIFN-alpha14, HYBRID 1 or HYBRID 2 for 24 hours at 37° C. in an atmosphere of 5% CO.sub.2 in air in a humidified incubator. IFN concentrations used were 0, 1, 5, 10, 50, 100, 1,000, 10,000, 100,000 and 1,000 ,000 IU/ml. Each IFN concentration (1ml) was cultured in triplicate in 12 well plates.

[0145] After 24 hours incubation, the supernatants were harvested into individual labelled microcentrifuge tubes and centrifuged at 11,000 RPMI for 5 minutes in a microcentrifuge. Samples are then collected and stored in labelled microcentrifuge tubes at -20° C. for assaying by ELISA.

Typical ELISA Procedure

[0146] One day prior to the beginning of the assay, a 96 well flat-bottomed ELISA plate was coated with 100 .Math.l of the corresponding Capture Antibody, as per the manufacturer’s instructions.

[0147] The following day, ELISA reagents and supernatants were allowed to heat to room temperature, and the coated ELISA plate was washed four times with wash buffer (1x phosphate buffer saline + 0.05%(v/v) Tween 20, and 200 .Math.l assay diluent is added to each well to prevent nonspecific antibody binding, and incubated at room temperature for 1 hour, with shaking (500 rpm).

[0148] The plate was washed 4 times, and 100 .Math.l of diluted standards and samples are added the appropriate wells. The plate is then sealed and incubated for 2 hours with shaking.

[0149] After a further 4 wash cycles, 100 ul of Detection Antibody was added to each well, followed by a further 1 hour incubation at room temperature, with shaking. The plate was washed a further 4 times, and diluted Avidin-HRP solution was added to each well, followed by a 30 minute incubation at room temperature, with shaking. The plate was then washed a total of 5 times, allowing for 30 seconds to 1 minute of soaking between washes, before adding 100 .Math.l Substrate Solution for 15 minutes in darkness.

[0150] After 15-45 minutes, or when the standard wells reach the desired colour, 100 .Math.l of stop solution (1M H.sub.2SO.sub.4) and the plate was read via spectrophotometry, typically at wavelengths 450 and 570 nm.

Results

[0151] The results obtained are shown in FIGS. 3 to 5. FIG. 3 demonstrates that IFNα-14 inhibits IL31 production in human leucocytes. FIG. 3 indicates strong inhibition of IL31 in the presence of IFNα-14 and in particular at low concentrations of IFNα-14.

[0152] FIG. 4 demonstrates that HYBRID 1 inhibits IL31 production in human leucocytes. FIG. 4 indicates strong inhibition of IL31 in the presence of HYBRID 1 and in particular at low concentrations of HYBRID 1.

[0153] FIG. 5 demonstrates that HYBRID 2 inhibits IL31 production in human leucocytes. FIG. 5 indicates strong inhibition of IL31 in the presence of HYBRID 2 and in particular at low concentrations of HYBRID 2.

[0154] By way of comparison, the inventors used Human Alpha-Interferon-2a (Roferon) to demonstrate that it would be unexpected for an interferon alpha subtype to inhibit IL31. FIG. 6 demonstrates a lack of suppression of IL-31 production by Human Alpha-Interferon-2a (Roferon) in human leukocytes.

[0155] These results surprisingly demonstrate that administration of IFN-α14, HYBRID 1 or HYBRID 2 results in a greater reduction or inhibition of IL31 in keratinocytes compared to previous medications.

[0156] 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 preferred 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.