Inhibitors

Abstract

The present invention relates to the use of an inhibitor of potassium channel Kv1.3 in a therapeutic method, in particular wherein said method comprises administering said inhibitor to a subject at specific intervals.

Description

Technical Field

[0001] The present invention relates to the use of inhibitors of the potassium channel Kv1.3 in therapeutic methods, in particular, wherein said methods comprise administering said inhibitor to a subject at specific intervals. Background Art

[0002] Ion channels are membrane proteins that form pores in biological membranes to allow (and regulate) the flow of ions across the associated membrane. There are many different types of ion channels, which can be classified in a variety of ways, such as by the type of ion for which they provide a channel, the way in which they regulate or "gate" the ion channel (e.g., "ligand-gated" or "voltage-gated"), and their cellular or subcellular localization.

[0003] Potassium channels are divided into four major classes, namely voltage-gated potassium channels, calcium-activated potassium channels, inwardly rectifying potassium channels, and tandem pore domain potassium channels.

[0004] Voltage-gated potassium channels, like other voltage-gated channels, open or close in response to transmembrane voltage. They represent a complex family with a variety of biological functions, including regulation of neurotransmitter release, heart rate, insulin secretion, neuronal excitability, epithelial electrolyte transport, smooth muscle contraction, and cell volume.

[0005] The Kv1.3 (voltage-gated potassium channel subfamily A member 3) channel is expressed on T cells and plays a role in regulating T cell activation. Inhibitors of Kv1.3 have been shown to inhibit the proliferation of activated T cells in vitro (reviewed in Cahalan and Chandy, Immunol. Rev. 231:59-87, 2009), and to inhibit T cell-dependent disease progression in multiple experimental models of autoimmune diseases, including experimental autoimmune encephalomyelitis (EAE), experimental arthritis, delayed-type hypersensitivity (DTH), allergic contact dermatitis, and glomerulonephritis. See, e.g., Rangaraju et al. (Expert Opin. Ther. Targets 13:909-24, 2009); Beeton et al. (Proc. Natl. Acad. Sci. U S A. 103:17414-9, 2006); Koo et al. (J. Immunol. 158:5120-8, 1997); Hyodo et al. (Am. J. Physiol. Renal Physiol. 299:F1258-69, 2010). WO 2016 / 112208 describes the topical application of Kv1.3 blockers for the treatment of skin and mucosal inflammation.

[0006] Inhibitors of Kv1.3 have been shown to inhibit the proliferation of activated T cells and to have beneficial effects in multiple disease experimental models. Without wishing to be bound by theory, it is believed that potassium efflux through the Kv1.3 channel in cells is required to maintain the calcium influx necessary for T cell activation.

[0007] Kv1.3 is overexpressed in: Gad5 / insulin-specific T cells from patients with new-onset type 1 diabetes, myelin-specific T cells from MS patients, and T cells from the synovium of rheumatoid arthritis patients (Beeton et al., Proc Natl Acad Sci USA 103:17414-9, 2006), breast cancer samples (Abdul et al., Anticancer Res 23:3347, 2003), and prostate cancer cell lines (Fraser et al., Pflugers Arch 446:559, 2003).

[0008] Positive results of using Kv1.3 inhibitors in animal models have been described in the following models: models of hypersensitivity to ovalbumin and tetanus toxoid (Beeton et al., Mol Pharmacol 67:1369, 2005; Koo et al., Clin Immunol 197:99, 1999), models for multiple sclerosis such as the rat adoptive-transfer experimental autoimmune encephalomyelitis (AT-EAE) model (Beeton et al., Proc Natl Acad Sci USA 103:17414-9, 2006), an inflammatory bone resorption model (Valverde et al., J Bone Mineral Res 19:155, 2004), arthritis models (Beeton et al., Proc Natl Acad Sci 103:17414, 2006; Tarcha et al., J. Pharmacol. Exp. Ther. 342:642, 2012), and obesity, diabetes and metabolic disorders (Xu et al., Hum Mol Genet 12:551, 2003; Xu et al., Proc Natl Acad Sci 101:3112, 2004). Topical application of Kv1.3 blockers has been proposed for the treatment of skin and mucosal inflammation.

[0009] Accordingly, Kv1.3 inhibitors have considerable potential for the treatment of diseases and disorders, particularly inflammatory disorders such as autoimmune diseases.

[0010] WO 2015 / 013330 proposes the use of Kv1.3 blocker peptides for the treatment of eye disorders such as dry eye and uveitis, including when caused by autoimmune diseases (such as Sjogren’s syndrome).

[0011] Kv1.3 inhibitors can also have beneficial metabolic effects, such as those related to energy homeostasis, body weight regulation, and glucose control. Compared to control littermates, Kv1.3 knockout (Kv1.3(- / -)) mice exhibit reduced weight gain, higher insulin sensitivity, and lower plasma glucose levels in response to a high-fat diet (Xu et al., Hum. Mol. Genet. 12:551-9, 2003). In addition, Kv1.3 inhibitors have also been shown to increase the expression of glucose transporter 4 (GLUT4) in skeletal muscle and adipose tissue, improve insulin sensitivity in normal and ob / ob obese mice, and enhance glucose uptake in primary adipocytes in vitro (Xu et al., Proc. Natl. Acad. Sci. USA 101:3112-7, 2004). In humans, single nucleotide polymorphisms (SNPs) in the Kv1.3 gene are also associated with reduced insulin sensitivity and impaired glucose tolerance (Tschritter, Clin Endocrinol Metab 91:654-8, 2006).

[0012] Kv1.3 is also expressed in proliferating human and mouse smooth muscle cells. Inhibitors of Kv1.3 can be effective against smooth muscle proliferative disorders such as restenosis (e.g., in patients who have undergone vascular surgery such as angioplasty). Kv1.3 inhibitors have been shown to inhibit calcium entry, reduce smooth muscle cell migration, and inhibit neointimal hyperplasia in ex vivo human vein samples (Cheong et al., Cardiovasc. Res. 89:282-9, 2011).

[0013] Additional evidence suggests that Kv1.3 channels are involved in the activation and / or proliferation of various types of cells, including tumor cells (Bielanska et al., Curr. Cancer Drug Targets 9:904-14, 2009), microglia (Khanna et al., Am. J. Physiol. Cell Physiol. 280:C796-806, 2001), and the differentiation of neuronal progenitor cells (Wang et al., J. Neurosci. 30:5020-7, 2010). Thus, Kv1.3 inhibitors may be beneficial for the treatment of neuroinflammatory and neurodegenerative disorders as well as cancer.

[0014] Kv1.3 is part of a closely related subfamily of potassium channels (referred to as Kv1.1 to Kv1.8). When dealing with large homologous families, it is always desirable for an inhibitor to be as selective and specific as possible for the desired target in order to improve potency and safety and to avoid unwanted off-target effects. The most specific Kv1.3 inhibitors identified to date are venom peptides derived from various types of venomous organisms such as snakes, arachnids (e.g., scorpions and spiders), sea anemones, etc. Such Kv1.3 inhibitors include the peptides ShK, Oskl, margatoxin, and kaliotoxin (reviewed by Chandy et al., Trends in Pharmacol. Sci. 25:280-9, 2004). See also Abdel-Mottaleb et al., Toxicon 51:1424-30, 2008 and Mouhat et al., Biochem. J. 385 (Pt 1):95-104, 2005.

[0015] Numerous attempts have been described to engineer toxin peptides for specific properties including specificity or potency, such as in WO2006 / 002850, WO2006 / 042151, WO2008 / 088422, WO2006 / 116156, WO2010 / 105184, and WO2014 / 116937. Summary of the Invention

[0016] The present invention relates to a Kv1.3 inhibitor which is a toxin peptide derived from the scorpion Parabuthus transvaalicus. The toxin peptide has the amino acid sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1).

[0017] Among other desirable properties, it has been found that this molecule and its derivatives or variants are highly selective inhibitors of the Kv1.3 potassium channel compared to other voltage-gated potassium channels and generally also have high potency in blocking the Kv1.3 channel. Peptides that are highly selective for the Kv1.3 channel, such as peptides derived from PaT1, have the particular advantage of selectively targeting cells that express the Kv1.3 channel (e.g., a specific subset of effector memory T cells). Thus, this selectivity confers the potential for Kv1.3 inhibitors derived from PaT1 to have a targeted therapeutic effect.

[0018] Kv1.3 inhibitors based on the PaT1 toxin peptide have a short half-life in vivo in a subject. For example, Example 6 herein shows that such a Kv1.3 inhibitor has a half-life of about 1 hour in a rat model organism. However, the inventors have unexpectedly found that despite this short half-life, the Kv1.3 inhibitors described herein also have a lasting effect on T cells. In particular, Example 8 herein shows that inflammation in a rat ear inflammation model was reduced for up to 7 days after treatment with a Kv1.3 inhibitor. Additionally, Example 9 herein shows that treatment with a Kv1.3 inhibitor once every 5 days reduced inflammation in a rat arthritis model. Without wishing to be bound by theory, it is hypothesized that the Kv1.3 inhibitor somehow "reprograms" T cells through its interaction with the Kv1.3 ion channel to remain inactive for an extended period of time, thereby suppressing inflammation for multiple days.

[0019] These unexpected findings support the concept of treating a subject having a disease, disorder, or condition treatable with a Kv1.3 inhibitor as described herein according to a regimen of administering the inhibitor at approximately weekly intervals (i.e., once every 2 to 8 days). It is advantageous to extend the time period between administrations of the active compound as much as possible because this saves time and effort (since fewer administrations are required within a given time period) and cost (since less of the active compound is required within a given time period). Especially in the case of administering the active compound by certain routes (e.g., subcutaneous injection), a longer interval between administrations also improves patient comfort and can thereby improve patient compliance with the treatment regimen. For example, for the reasons described above, administering a Kv1.3 inhibitor according to the invention (i.e., once every 2 to 8 days) is superior to daily administration of the inhibitor. The unexpected finding described in the present disclosure is that despite the short in vivo half-life of the Kv1.3 inhibitor, it is possible to administer the Kv1.3 inhibitor described herein once every 2 to 8 days because an effect of the inhibitor (e.g., reduction of inflammation) was observed for multiple days after administration.

[0020] Accordingly, the present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use in a method of treating or preventing a disease or disorder in a subject, wherein the Kv1.3 inhibitor comprises or consists of a peptide comprising the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof, or consists of the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof, wherein the variant (a) has at least 65% sequence identity with SEQ ID NO 1, and / or (b) differs from SEQ ID NO 1 by a total of at most nine substitutions, insertions and / or deletions, and wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Depicts the study design of a keyhole limpet hemocyanin (KLH)-induced delayed type hypersensitivity (DTH) model to study the effect of a Kv1.3 inhibitor (peptide 100) on ear swelling after ear challenge on days 7, 9, 11 or 13. Ear edema of respective ear challenges was measured 24 or 48 hours after challenge.

[0022] Figure 2 Shows the effect of peptide 100 administered at 300 nmol / kg on day 6 on ear swelling after ear challenge on days 7, 9, 11 or 13, measured 24 hours after challenge. Data are shown as individual values and mean (n = 8 rats / group). Means of vehicle- and peptide 100-treated animals at respective ear challenge time points were compared by two-tailed unpaired t-test.

[0023] Figure 3 Shows the effect of peptide 100 administered at 300 nmol / kg on day 6 on ear swelling after ear challenge on days 7, 9, 11 or 13, measured 48 hours after challenge. Data are shown as individual values and mean (n = 8 rats / group). Means of vehicle- and peptide 100-treated animals at respective ear challenge time points were compared by two-tailed unpaired t-test.

[0024] Figure 4 Shows the clinical score of the front paws in a rat collagen-induced arthritis (CIA) model described in Example 9.

[0025] Figure 5Depicts the study design of a keyhole limpet hemocyanin (KLH)-induced delayed type hypersensitivity (DTH) model to study the effect of different doses of the Kv1.3 inhibitor (peptide 100) on ear swelling after ear challenge on day 7 or 11. Ear edema of the respective ear challenges was measured 24 hours or 48 hours after the challenge.

[0026] Figure 6 Shows the effect of peptide 100 administered at 10, 100, 300, or 700 nmol / kg on day 6 on ear swelling after ear challenge on day 7 or 11, measured 24 hours after the challenge. Data are shown as individual values and mean (n = 8 rats / group). The means of vehicle- and peptide 100-treated animals were compared at the respective ear challenge time points by two-sided unpaired t-test.

[0027] Figure 7 Shows the effect of peptide 100 administered at 10, 100, 300, or 700 nmol / kg on day 6 on ear swelling after ear challenge on day 7 or 11, measured 48 hours after the challenge. Data are shown as individual values and mean (n = 8 rats / group). The means of vehicle- and peptide 100-treated animals were compared at the respective ear challenge time points by two-sided unpaired t-test.

[0028] Figure 8 Shows the effect of peptide 100 administered at 1, 3, 10, 30, or 100 nmol / kg on day 6 on ear swelling after ear challenge on day 7 or 11, measured 24 hours after the challenge. Data are shown as individual values and mean (n = 8 rats / group). The means of vehicle- and peptide 100-treated animals were compared at the respective ear challenge time points by two-sided unpaired t-test.

[0029] Figure 9 Shows the effect of peptide 100 administered at 1, 3, 10, 30, or 100 nmol / kg on day 6 on ear swelling after ear challenge on day 7 or 11, measured 48 hours after the challenge. Data are shown as individual values and mean (n = 8 rats / group). The means of vehicle- and peptide 100-treated animals were compared at the respective ear challenge time points by two-sided unpaired t-test. Detailed Description

[0030] The present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use in a method of treating or preventing a disease or disorder in a subject, wherein the Kv1.3 inhibitor comprises or consists of a peptide comprising the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO: 1) or a variant thereof or consisting of the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO: 1) or a variant thereof, wherein the variant (a) has at least 70% sequence identity with SEQ ID NO: 1, and / or (b) differs from SEQ ID NO: 1 by a total of at most nine substitutions, insertions and / or deletions, and wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

[0031] The peptide can be any peptide described herein. The disease or disorder can be any disease or disorder described herein. Kv1.3 inhibitor

[0032] The present invention provides a Kv1.3 inhibitor that comprises or consists of a peptide.

[0033] The term "Kv1.3 inhibitor" is used herein to denote a molecule or compound that has inhibitor (or blocking) activity against the Kv1.3 ion channel, i.e., is capable of inhibiting or eliminating ion flow through the Kv1.3 ion channel, e.g., by binding to the ion channel. The term "blocker" as used herein is synonymous with the term "inhibitor". Thus, the Kv1.3 inhibitor of the present invention may also be referred to herein as an "ion channel blocker". The terms "blocker" and "inhibitor" should not be construed as implying any particular mechanism of action or any particular mode of interaction with the ion channel itself.

[0034] The terms "Kv1.3 inhibitor" and "compound" are used interchangeably herein.

[0035] The term "Kv1.3" is used to refer to potassium voltage-gated channel subfamily A member 3, also known as KCNA3, HPCN3, HGK5, HuKIII, and HLK3. "Subfamily A" may also be referred to as the "oscillator-related subfamily". The human amino acid sequence of Kv1.3 is provided under UniProt accession number P22001, version P22001.3 (Q5VWN2).

[0036] The Kv1.3 channel is expressed on T and B lymphocytes and is associated with T cell activation. Many research groups are developing Kv1.3 blockers for suppressing immune responses and for a variety of other indications. However, the Kv1.3 channel is part of a complex family of related ion channels, which also includes Kv1.1, Kv1.2, and Kv1.6 channels with different physiological roles. Thus, it is desirable that Kv1.3 inhibitors be as selective for Kv1.3 as possible over other ion channels, especially other voltage-gated potassium channels such as Kv1.1, Kv1.2, Kv1.4, Kv1.5, Kv1.6, Kv1.7, and Kv1.8. More information on Kv1.3 and known Kv1.3 inhibitors can be found in Murray et al. J Med Chem 2015, 58, 17, 6784 - 6802 and Tanner et al. Clin Immunol 2017, 180, 45 - 47.

[0037] The Kv1.3 inhibitors or pharmaceutically acceptable salts thereof of the present invention have Kv1.3 inhibitor activity. In other words, the Kv1.3 inhibitors of the present invention (and the peptide components of the isolated Kv1.3 inhibitors) have inhibitor or blocker activity at the Kv1.3 ion channel, i.e., it is capable of inhibiting ion flow through the Kv1.3 channel. IC 50 value

[0038] IC 50 values can be used as a measure of inhibitor (or blocker) activity or potency. IC 50 values are a measure of the inhibitor concentration required to achieve half-maximal inhibition of ion channel activity in a given assay. A compound having a lower IC 50 under a particular ion channel than a reference compound can be considered a more active inhibitor or a more potent inhibitor than the reference compound. The terms "activity" and "potency" are used interchangeably.

[0039] IC 50 values can be determined using any suitable assay, such as fluorescence-based assays measuring ion flux (e.g., thallium ion flux) and patch-clamp assays, which can be carried out as described in the examples herein. Patch-clamp assays may be preferred, for example using systems.

[0040] In some embodiments, the Kv1.3 inhibitors of the present invention have an IC 50is about 400 nM or less, such as about 300 nM or less, such as about 200 nM or less, such as about 100 nM or less, such as about 50 nM or less, such as about 15 nM or less, such as about 10 nM or less, such as about 5 nM or less. Preferably, the IC 50 of the Kv1.3 inhibitor of the present invention is about 2 nM or less. More preferably, the IC 50 of the Kv1.3 inhibitor of the present invention is about 1 nM or less, such as about 0.5 nM or less. Selectivity

[0041] The Kv1.3 inhibitor of the present invention is selective for Kv1.3. In some embodiments, the Kv1.3 inhibitor of the present invention is selective for Kv1.1, Kv1.2, Kv1.4, Kv1.5, Kv1.6, Kv1.7, and Kv1.8. In particular, the Kv1.3 inhibitor of the present invention is selective for Kv1.3 over one or more of Kv1.1, Kv1.2, and Kv1.6.

[0042] For example, the Kv1.3 inhibitor of the present invention may be selective for Kv1.3 over Kv1.1, selective for Kv1.3 over Kv1.2, selective for Kv1.3 over Kv1.6, selective for Kv1.3 over Kv1.1 and Kv1.2, selective for Kv1.3 over Kv1.1 and Kv1.6, selective for Kv1.3 over Kv1.2 and Kv1.6, or selective for Kv1.3 over Kv1.1, Kv1.2, and Kv1.6. Generally, the Kv1.3 inhibitor is selective for Kv1.3 over Kv1.1. Additionally, its selectivity for Kv1.3 may be over Kv1.2 and / or Kv1.6.

[0043] "Selectivity" herein means that the inhibitor activity of the Kv1.3 inhibitor against Kv1.3 is higher than the inhibitor activity against each of Kv1.1, Kv1.2, and Kv1.6. Thus, the IC 50 of the Kv1.3 inhibitor against Kv1.3 is generally lower than the IC 50 against each of the corresponding other one or more ion channels.

[0044] Therefore, the selectivity for Kv1.3 over another ion channel X can be expressed as the ratio of the respective IC 50 values, for example, expressed as IC 50 [X] / IC 50 [Kv1.3].

[0045] The Kv1.3 inhibitors of the present invention can be selective for Kv1.3 over Kv1.1 by at least about 10, at least about 100, at least about 1000, or at least about 10000, and can be up to about 100000 or even higher. Generally, the Kv1.3 inhibitors of the present invention are selective for Kv1.3 over Kv1.1 by at least about 100, or at least about 1000.

[0046] The Kv1.3 inhibitors of the present invention can be selective for Kv1.3 over Kv1.2 by at least about 10, at least about 100, at least about 1000, or at least about 10000, and can be up to about 100000 or even higher. Generally, the inhibitor is selective for Kv1.3 over Kv1.2 by at least 10, and preferably at least about 50, or at least about 100, or at least about 1000.

[0047] The Kv1.3 inhibitors of the present invention can be selective for Kv1.3 over Kv1.6 by at least about 10, at least about 100, at least about 1000, or at least about 10000, and can be up to about 100000 or even higher. Generally, the inhibitor is selective for Kv1.3 over Kv1.6 by at least 100, or at least about 400, or at least about 1000.

[0048] The Kv1.3 inhibitors of the present invention can have higher selectivity than known Kv1.3 inhibitors such as ShK, Mokatoxin (Moka1), Vm24, Odk2, or Osk1. Thus, the Kv1.3 inhibitors of the present invention can have higher selectivity for Kv1.3 over ion channel X, i.e., IC 50 [X] / IC 50 [Kv1.3], which is greater than the selectivity of the comparative molecule. The selectivity of the two Kv1.3 inhibitors will be determined for each ion channel under the same conditions for direct comparison. As described above, any suitable assay can be used, such as fluorescence-based ion flux assays and patch clamp assays.

[0049] In the case of any one or all of Kv1.1, Kv1.2, and / or Kv1.6, the Kv1.3 inhibitors of the present invention can have lower absolute inhibitor activity (i.e., higher IC 50 ) than known Kv1.3 inhibitors (such as Odk2 or Osk1). However, it is acceptable that: as long as the Kv1.3 inhibitors of the present invention are more selective for Kv1.3 than the comparative compound, then in the case of any one or all of these ion channels, the Kv1.3 inhibitors of the present invention have lower absolute inhibitor activity. Generally, the Kv1.3 inhibitors of the present invention combine high specificity and high potency for Kv1.3. Pharmaceutically acceptable salts

[0050] The Kv1.3 inhibitor of the present invention may be in the form of a pharmaceutically acceptable salt. All references herein to "Kv1.3 inhibitor", "the Kv1.3 inhibitor of the present invention", "peptide" or "the peptide of the present invention" shall be considered to cover any pharmaceutically acceptable salt thereof, whether or not "pharmaceutically acceptable salt" is expressly recited. The Kv1.3 inhibitor may also be referred to as a "solvate", which means a defined stoichiometric complex formed between a solute (the Kv1.3 inhibitor of the present invention or its pharmaceutically acceptable salt) and a solvent. In this regard, the solvent may be, for example, water, ethanol or some other pharmaceutically acceptable organic substance, such as a small molecule organic substance, such as, but not limited to, acetic acid or lactic acid. When the solvent in question is water, such a solvate is generally referred to as a hydrate. In some embodiments, the pharmaceutically acceptable salt of the present invention is an acetate. In other words, the present invention encompasses a salt comprising a cation of a Kv1.3 inhibitor and an acetate anion or consisting of a cation of a Kv1.3 inhibitor and an acetate anion. In some embodiments, the pharmaceutically acceptable salt of the present invention is a chloride salt. In other words, the present invention encompasses a salt comprising a cation of a Kv1.3 inhibitor and a chloride anion or consisting of a cation of a Kv1.3 inhibitor and a chloride anion. Peptide

[0051] The Kv1.3 inhibitor of the present invention comprises or consists of such a peptide, which peptide comprises the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof or consists of the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof, wherein the variant (a) has at least 70% sequence identity with SEQ ID NO 1, and / or (b) differs from SEQ ID NO 1 by a total of at most nine substitutions, insertions and / or deletions.

[0052] SEQ ID NO 1 is the amino acid sequence of a toxin peptide of the scorpion Androctonus australis. As described herein, this peptide is a selective inhibitor of the Kv1.3 potassium ion channel. Thus, it is the peptide moiety of the Kv1.3 inhibitor of the present invention that is considered to inhibit Kv1.3. This peptide may be referred to herein as the "peptide component", "peptide element", "Kv1.3 inhibitor component" or "Kv1.3 inhibitor element" of the Kv1.3 inhibitor of the present invention.

[0053] Variants of SEQ ID NO:1 are peptides that contain one or more amino acids that are different from, additional to, or missing relative to the amino acids of SEQ ID NO:1. In other words, the variant contains one or more amino acid changes compared to SEQ ID NO:1. Such variants may also be referred to herein as "derivatives", "variant sequences", "sequence variants", "variant peptides", "peptide variants", or simply "peptides".

[0054] In some embodiments, the Kv1.3 inhibitors of the invention comprise a peptide as described herein. In other words, in some embodiments, the Kv1.3 inhibitor comprises the peptide and other features or elements. In some embodiments, the Kv1.3 inhibitors of the invention consist of a peptide as described herein. In other words, in some embodiments, the Kv1.3 inhibitor is a peptide (i.e., the Kv1.3 inhibitor consists of a peptide and has no other features or elements).

[0055] In some embodiments, the peptide consists of the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO:1) or a variant thereof, wherein the variant (a) has at least 70% sequence identity with SEQ ID NO:1, and / or (b) differs from SEQ ID NO:1 by a total of at most nine substitutions, insertions, and / or deletions. Amino acid

[0056] Throughout this specification and the claims, the conventional three-letter and single-letter codes for naturally occurring amino acids are used, namely A (Ala), G (Gly), L (Leu), I (Ile), V (Val), F (Phe), W (Trp), S (Ser), T (Thr), Y (Tyr), N (Asn), Q (Gln), D (Asp), E (Glu), K (Lys), R (Arg), H (His), M (Met), C (Cys), and P (Pro); and the well - recognized codes for other α - amino acids, such as sarcosine (Sar), norleucine (Nle), α - amino - isobutyric acid (Aib), 2,3 - diamino - propionic acid (Dap), 2,4 - diamino - butyric acid (Dab), 2,5 - diamino - valeric acid (ornithine; Orn), α - amino - butyric acid (Abu, also known as homo - alanine), hK, hLys or homo - Lys (homo - lysine), hQ, hGln or homo - Gln (homo - glutamine, also known as 6 - oxo - lysine, L - 5 - carbamoyl - norvaline, 6 - amino - 6 - oxo - norleucine or 5 - (aminocarbonyl) - norvaline), F(4 - F) (4 - fluoro - phenylalanine), F(4 - NH2) (4 - amino - phenylalanine), F(4 - NO2) (4 - nitro - phenylalanine), F(4 - CH3) (4 - methyl - phenylalanine).

[0057] The name [2 - amino - 5 - carboxy - pentanoyl] represents the peptide residue of 2 - amino - 5 - carboxy - pentanoic acid: Thus, it has a side chain similar to that of glutamic acid, but with an additional methylene group.

[0058] The name [2,3 - diamino - propionyl] represents the peptide residue of 2,3 - diamino - propionic acid, which has the following structure:

[0059] The name [2,4 - diamino - butyryl] represents the peptide residue of 2,4 - diamino - butyric acid, which has the following structure:

[0060] The name [2 - amino - 3 - guanidino - propionyl] represents the peptide residue of 2 - amino - 3 - guanidino - propionic acid, which has the following structure:

[0061] When used in the general formulas or sequences in this specification, especially when the rest of the formula or sequence is shown using single - letter codes, such additional α - amino acids can be shown in square brackets “[]” (e.g., “[Nle]”). Unless otherwise specified, the amino acid residues in the peptides of the present invention have the L - configuration. However, D - configuration amino acids can be incorporated. In the context of the present invention, amino acid codes represented by lowercase letters represent the D - configuration of the said amino acids, e.g., “k” represents the D - configuration of lysine (K). Numbering of amino acid positions

[0062] The amino acid residues of SEQ ID NO 1 are numbered from 1 to 37 in the conventional direction from the N-terminus to the C-terminus. Throughout the specification, the amino acid positions in variants of SEQ ID NO 1 are numbered according to the corresponding positions in SEQ ID NO 1 when optimally aligned with SEQ ID NO 1. Thus, particularly for variants that contain one or more insertions or deletions compared to SEQ ID NO 1, the numbering of any given residue reflects the corresponding residue in SEQ ID NO 1, and does not necessarily reflect its linear position in the variant sequence.

[0063] Residues at a particular position can be represented by the number of the relevant position together with the one-letter code or three-letter code of the residue present. Thus, 1Q or Q1 (the two forms are interchangeable) represents the glutamine (Q) residue at position 1, while 2Nle, 2[Nle], Nle2 or [Nle]2 represents the norleucine residue at position 2.

[0064] An asterisk can be used to denote a deleted position relative to the SEQ ID NO 1 sequence. For example, "1*" indicates that the residue at position 1 is deleted compared to SEQ ID NO 1.

[0065] An insertion can be represented by a string of consecutive residues at a single position. For example, "1QA" represents the insertion of an alanine (A) residue after the glutamine (Q) residue at position 1.

[0066] In some embodiments, any substitution compared to SEQ ID NO 1 is a conservative substitution. However, any substitution listed in any of the general formulas provided below can be introduced at the corresponding position. Cysteine and disulfide bonds

[0067] The peptide contains six cysteine (C) residues that together form three disulfide bonds between residue 6C and 27C, between residue 12C and 32C, and between residue 16C and 34C. Thus, in the Kv1.3 inhibitors or pharmaceutically acceptable salts of the present invention, the peptide contains cysteine (C) at each of positions 6, 12, 16, 27, 32, and 34.

[0068] By reference to SEQ ID NO 1, the disulfide bonds can be represented visually as follows: QMDMRC(1)SASVEC(2)KQKC(3)LKAIGSIFGKC(1)MNKKC(2)KC(3)YPR Pairs of cysteine residues that participate in a disulfide bond together are denoted by the same number in parentheses. Similar notations can be applied to any other sequences in this application. Unless the context requires otherwise, it should be understood that the active inhibitor compound contains the appropriate disulfide bond. In some embodiments, the pattern of the disulfide bridge is different from the pattern shown in SEQ ID NO 1 above. In such embodiments, the different pattern of the disulfide bridge is represented by the above numbering system.

[0069] It may be desirable that no other cysteine residues are introduced into the variant of SEQ ID NO 1 by substitution or insertion. Thus, in some embodiments, the variant does not contain other cysteine residues except those at positions corresponding to positions 6, 12, 16, 27, 32, and 34 of SEQ ID NO 1. In some embodiments, any substitution or deletion in the variant of SEQ ID NO 1 is not at the amino acids at positions 6, 12, 16, 27, 32, and 34 of SEQ ID NO 1. Sequence identity

[0070] In some embodiments, the variant has at least 70% sequence identity with SEQ ID NO 1.

[0071] The sequence of the variant of the Kv1.3 inhibitor of the present invention can be represented by sequence identity rather than by the number of substitutions, insertions, and deletions relative to SEQ ID NO 1.

[0072] Accordingly, the present invention provides a method for using a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof in the treatment or prevention of a disease or disorder in a subject, wherein the Kv1.3 inhibitor comprises or consists of a peptide that comprises the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof, and wherein the variant has at least 70% sequence identity with SEQ ID NO 1, and wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

[0073] In some embodiments, the peptide has at least 75% sequence identity with SEQ ID NO:1, such as at least 80% sequence identity with SEQ ID NO:1, such as at least 85% sequence identity with SEQ ID NO:1, such as at least 90% sequence identity with SEQ ID NO:1, such as at least 95% sequence identity with SEQ ID NO:1, such as at least 96% sequence identity with SEQ ID NO:1, such as at least 97% sequence identity with SEQ ID NO:1, such as at least 98% sequence identity with SEQ ID NO:1, such as at least 99% sequence identity with SEQ ID NO:1, such as 100% sequence identity with SEQ ID NO:1.

[0074] In some embodiments, the "percent (%) sequence identity" of a peptide is defined as the percentage of amino acids in the peptide sequence that are identical to the amino acids in the wild-type toxin peptide sequence SEQ ID NO 1 after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity and without regard to any conservative substitutions that are part of the sequence identity. Sequence alignments can be performed by one of ordinary skill in the art using techniques well known in the art, such as using publicly available software (e.g., BLAST, BLAST2, or Align software). See, for example, Altschul et al., Methods in Enzymology 266:460-480 (1996) or Pearson et al., Genomics 46:24-36, 1997.

[0075] The percent sequence identity used herein can be determined using these programs with their default settings in the context of the present invention. More generally, one of ordinary skill in the art can readily determine the appropriate parameters for determining the alignment, including any algorithms required to achieve the maximum alignment over the full length of the sequences being compared. Substitutions, insertions, and deletions

[0076] Variants of SEQ ID NO 1 differ from SEQ ID NO 1 by the deletion of one or more amino acids of SEQ ID NO 1 and / or the replacement of one or more amino acids of SEQ ID NO 1 with different amino acids and / or the insertion of one or more amino acids into the sequence of SEQ ID NO 1. Amino acids can be inserted at internal positions of SEQ ID NO 1, at the N-terminus of SEQ ID NO 1, or at the C-terminus of SEQ ID NO 1. Thus, variants of SEQ ID NO 1 contain one or more substitutions, insertions, and / or deletions.

[0077] Unless otherwise indicated, "substitution" refers to the replacement (i.e., substitution) of a single amino acid in SEQ ID NO 1. Thus, for example, the substitution of three consecutive amino acids in SEQ ID NO 1 constitutes three substitutions, not a single substitution. Similarly, "insertion" refers to the insertion of a single amino acid into SEQ ID NO 1 (which can be internal, at the N-terminus, and / or at the C-terminus), and thus, for example, the insertion of three consecutive amino acids into SEQ ID NO 1 constitutes three insertions, not a single insertion. "Deletion" refers to the deletion of a single amino acid from SEQ ID NO 1, and thus, for example, the deletion of three consecutive amino acids from SEQ ID NO 1 constitutes three deletions, not a single deletion.

[0078] In some embodiments, the variant differs from SEQ ID NO 1 by a total of at most nine substitutions, insertions, and / or deletions. The term "a total of nine..." means that a total of at most nine amino acids can be substituted in SEQ ID NO 1, and / or inserted into SEQ ID NO 1, and / or deleted from SEQ ID NO 1. In other words, the maximum total combination of all substitutions, insertions, and deletions in the variant is nine, and within these nine can be any combination of substitutions, insertions, and / or deletions. In other words, the variant contains any combination of substitutions, insertions, and / or deletions with a maximum total combination of nine substitutions, insertions, and deletions.

[0079] The sequence of the variant of the Kv1.3 inhibitor of the present invention can be represented by the number of substitutions, insertions, and deletions relative to SEQ ID NO 1, rather than by sequence identity.

[0080] Accordingly, the present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use in a method of treating or preventing a disease or disorder in a subject, wherein the Kv1.3 inhibitor comprises or consists of a peptide comprising the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof, and wherein the variant differs from SEQ ID NO1 by a total of at most nine substitutions, insertions, and / or deletions, and wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

[0081] In some embodiments, the variant of SEQ ID NO 1 differs from SEQ ID NO 1 by at most a total of eight substitutions, insertions, and / or deletions. In some embodiments, the variant differs from SEQ ID NO 1 by at most a total of seven, at most six, at most five, at most four, at most three, or at most two substitutions, insertions, and / or deletions, or a total of one substitution, insertion, or deletion. In some embodiments, compared to SEQ ID NO 1, the variant contains a total of 9, 8, 7, 6, 5, 4, 3, 2, or 1 substitution, insertion, and / or deletion. Preferably, compared to the sequence of SEQ ID NO 1, the peptide contains a total of 6 substitutions, insertions, and / or deletions. Amino acid at a specific position

[0082] In some embodiments of the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention, the peptide comprises the following amino acids: The amino acid at position 1 is H, N, P, p, Q, S, V, or Y, or is absent; The amino acid at position 2 is I, M, or Nle, or is absent; The amino acid at position 3 is D, E, or S, or is absent; The amino acid at position 4 is E, L, M, Nle, S, or V, or is absent; The amino acid at position 5 is R or K, or is absent; The amino acid at position 7 is E, F, H, K, Orn, R, S, Y, 2,3-diaminopropionyl, 2,4-diaminobutyryl, or 2-amino-3-guanidinopropionyl; The amino acid at position 8 is A, H, I, L, S, or Y; The amino acid at position 9 is F, L, P, S, Orn, V, Abu, or 2,3-diaminopropionyl; The amino acid at position 10 is K, P, Q, R, or V; The amino acid at position 11 is E or Q; The amino acid at position 13 is A, E, G, K, L, Q, or V; The amino acid at position 14 is E, K, L, Q, V, or 2-amino-5-carboxypentanoyl; The amino acid at position 15 is K, L, P, S; The amino acid at position 17 is K, L, R, or Y, or is absent; The amino acid at position 18 is A, D, G, K, Q, hQ, V, or Y, or is absent; The amino acid at position 19 is A, K, R, or Y, or is absent; The amino acid at position 20 is E, I, R, or Y, or is absent; The amino acid at position 21 is E, G, H or R; The amino acid at position 22 is C, R or S; The amino acid at position 23 is G, I, K, P or R; The amino acid at position 26 is K or hK; The amino acid at position 28 is M or Nle; The amino acid at position 30 is G or K; The amino acid at position 33 is H, K, R or V; The amino acid at position 35 is Y, F(4-F), F(4-CH3), F(4-NO2) or F(4-NH2); The amino acid at position 36 is Q or P, or absent; and / or The amino acid at position 37 is C, G, R, S or (4-amino-5-hydroxypentyl)guanidine, or absent.

[0083] In some embodiments of the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention, the peptide comprises the following amino acids: The amino acid at position 1 is N or P, or absent; The amino acid at position 2 is M or Nle, or absent; The amino acid at position 3 is D or E, or absent; The amino acid at position 4 is M or Nle, or absent; The amino acid at position 5 is R, K, or absent; The amino acid at position 7 is S, 2,4-diaminobutyryl or 2-amino-3-guanidinopropionyl; The amino acid at position 14 is K or Q; The amino acid at position 18 is K or A; The amino acid at position 19 is K, S or A; The amino acid at position 28 is M or Nle; and / or The amino acid at position 37 is R or S.

[0084] In some preferred embodiments of the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention, the peptide comprises the following amino acids: The amino acid at position 1 is P; The amino acid at position 2 is Nle; The amino acid at position 3 is E; The amino acid at position 4 is Nle; The amino acid at position 18 is A; and / or The amino acid at position 28 is Nle.

[0085] In some preferred embodiments, positions 1 to 5 of the variant are deleted, or comprise or consist of an amino acid sequence selected from: QMDMR (SEQ ID NO:155), NMDMR (SEQ ID NO:156), P[Nle]D[Nle]R, and P[Nle]E[Nle]R.

[0086] In some embodiments, position 14 of the variant is K or Q. In some embodiments, position 28 of the variant is M or Nle. In some embodiments, position 37 of the variant is R or S. In some embodiments, the amino acid at position 1 of the variant of SEQ ID NO 1 is not Q. In some embodiments, the amino acid at position 1 of the variant of SEQ ID NO 1 is N or P, or is deleted. In some embodiments, the variant of SEQ ID NO 1 comprises amino acids 22S and 23I.

[0087] In some embodiments of the Kv1.3 inhibitors of the present invention, one or more positions of the variant of SEQ ID NO 1 are the same amino acid as the corresponding position in SEQ ID NO 1. In other words, the amino acid present at the relevant position is the same as the amino acid present at the corresponding position in SEQ ID NO 1.

[0088] In some embodiments, positions 6, 12, 16, 27, 32, and 34 of the variant are the same amino acid as the corresponding positions in SEQ ID NO 1. In other words, in some embodiments, the peptide comprises the following amino acids: 6C, 12C, 16C, 27C, 32C, and 34C.

[0089] In some embodiments, positions 6, 12, 16, 24, 25, 27, 32, and 34 of the variant are the same amino acid as the corresponding positions in SEQ ID NO 1. In other words, in some embodiments, the peptide comprises the following amino acids: 6C, 12C, 16C, 24F, 25G, 27C, 32C, and 34C.

[0090] In some embodiments, positions 6, 12, 16, 24, 25, 27, 29, 31, 32, and 34 of the variant are the same amino acid as the corresponding positions in SEQ ID NO1. In other words, in some embodiments, the peptide comprises the following amino acids: 6C, 12C, 16C, 24F, 25G, 27C, 29N, 31K, 32C, and 34C.

[0091] In some embodiments, the following positions of the variant are amino acids identical to the corresponding positions in SEQ ID NO 1: positions 6, 8 to 13, 15 to 17, 20 to 27, and 29 to 36. In some embodiments, the following positions of the variant are amino acids identical to the corresponding positions in SEQ ID NO 1: positions 6 to 13, 15 to 17, 20 to 27, and 29 to 36.

[0092] In some embodiments, any substitution or deletion in the variant of SEQ ID NO 1 is located at an amino acid position selected from positions 1 to 5, 7 to 11, 13 to 15, 17 to 23, 26, 28, 30, 33, and 35 to 37 of SEQ ID NO 1. Preferably, any substitution or deletion in the variant of SEQ ID NO 1 is located at an amino acid position selected from positions 1, 2, 3, 4, 5, 7, 14, 18, 19, 28, and 37 of SEQ ID NO 1.

[0093] In some embodiments, at least one amino acid in positions 7, 8, 9, 10, or 11 of SEQ ID NO:1 is replaced with an amino acid having a positively charged side chain and / or an amino acid having an aromatic side chain. In some embodiments, at least one amino acid in positions 7, 8, 9, or 10 of SEQ ID NO:1 is replaced with an amino acid having a positively charged side chain and / or an amino acid having an aromatic side chain. In some embodiments, exactly one amino acid in positions 7, 8, 9, or 10 of SEQ ID NO:1 is replaced with an amino acid having a positively charged side chain and / or an amino acid having an aromatic side chain. In some embodiments, the amino acids having a positively charged side chain are selected from H, R, Orn, 2,3-diaminopropionyl, 2-amino-3-guanidinopropionyl, and 2,4-diaminobutyryl. In some embodiments, the amino acid having an aromatic side chain is Y. Deletion

[0094] In some embodiments, if the variant of SEQ ID NO 1 contains one or more deletions, one of these deletions is located at position 1. In some embodiments, if the variant of SEQ ID NO 1 contains two or more deletions, two of these deletions are located at positions 1 and 2.

[0095] In some embodiments, the variant contains exactly 1, 2, 3, 4, 5, 6, or 7 deletions.

[0096] In some embodiments, the deletions in the peptide variant are selected from: Deletion at position 1; Deletions at positions 1 and 2; Deletions at positions 1, 2, and 3; Deletions at positions 1, 2, 3, and 4; Deletion of positions 1, 2, 3, 4, and 5; Deletion of positions 1, 2, 3, 4, 5, and 36; Deletion of positions 1, 2, 3, 4, 5, 36, and 37; Deletion of position 17; Deletion of position 18; Deletion of position 19; Deletion of position 19; Deletion of position 20; Deletion of position 36; and Deletion of position 37.

[0097] In some preferred embodiments, the deletion in the variant is at positions 1, 2, 3, 4, and 5. Insertion

[0098] In some embodiments, the variant contains up to four insertions compared to SEQ ID NO 1. In some embodiments, the variant contains up to three insertions, up to two insertions, or up to one insertion compared to SEQ ID NO 1.

[0099] In some embodiments, the peptide contains one or more insertions at the N-terminus (i.e., before position 1). In some embodiments, the peptide contains only one or more insertions at the N-terminus. In some embodiments, the peptide contains one or more insertions at the C-terminus (i.e., after position 37). In some embodiments, the peptide contains only one or more insertions at the C-terminus. In some embodiments, the peptide contains one or more insertions at both termini.

[0100] In some embodiments, the insertion at the N-terminus contains the sequence GG or SG or consists of the sequence GG or SG. In some embodiments, the C-terminal insertion contains the following or consists of the following: the sequence RRTA (SEQ ID NO:158), HRRK (SEQ ID NO:159), QSKA (SEQ ID NO:160), AGPR (SEQ ID NO:161), RSRT (SEQ ID NO:162), RHKR (SEQ ID NO:163), GGKR (SEQ ID NO:164), PKTA (SEQ ID NO:165), TDAR (SEQ ID NO:166), HRQQ (SEQ ID NO:167), RPRH (SEQ ID NO:168), ARNA (SEQ ID NO:169), TGRK (SEQ ID NO:170), HERT (SEQ ID NO:171), NTRT (SEQ ID NO:172), QRNG (SEQ ID NO:173), AHRN (SEQ ID NO:174), PRSA (SEQ ID NO:175), QRQS (SEQ ID NO:176), QRRK (SEQ ID NO:177), ARAK (SEQ ID NO:178), AKRD (SEQ ID NO:179), RDKT (SEQ ID NO:180), RAKR (SEQ ID NO:182), QRTR (SEQ ID NO:183), ATRH (SEQ ID NO:184), ARRS (SEQ ID NO:185), AKTR (SEQ ID NO:186), NRQR (SEQ ID NO:187) or PRNT (SEQ ID NO:188).

[0101] In some embodiments, the insertions in the variant are selected from: GG at positions -1 and 0 (i.e., inserted before position 1); SG at positions -1 and 0 (i.e., inserted before position 1); R at position 38 (i.e., inserted after position 37); Y at position 38 (i.e., inserted after position 37); L at position 38 (i.e., inserted after position 37); H at position 38 (i.e., inserted after position 37); E at position 38 (i.e., inserted after position 37); KS at positions 38 and 39 (i.e., inserted after position 37); FE at positions 38 and 39 (i.e., inserted after position 37); HR at positions 38 and 39 (i.e., inserted after position 37); AK at positions 38 and 39 (inserted after position 37); 4-amino-5-hydroxypentanamide at position 38 (inserted after position 37); ST at positions 38 and 39 (inserted after position 37); RY at positions 38 and 39 (inserted after position 37); RRTA (SEQ ID NO:158) at positions 38 to 41 (inserted after position 37); HRRK (SEQ ID NO:159) at positions 38 to 41 (inserted after position 37); and RRTK (SEQ ID NO:157) at positions 38 to 41 (inserted after position 37).

[0102] In some embodiments, the peptide is a fusion protein comprising SEQ ID NO1 or a variant thereof as defined herein, and one or more heterologous polypeptide sequences. In some embodiments, SEQ ID NO 1 or a variant thereof is inserted into a heterologous scaffold polypeptide. In some embodiments, the maximum length of the peptide is 200 amino acids, 150 amino acids, 125 amino acids, 100 amino acids, 75 amino acids or 50 amino acids. Terminal group

[0103] The “H” (or “Hy-”) portion at the N-terminus of the sequence represents a hydrogen atom, corresponding to the presence of a free primary or secondary amino group at the N-terminus. Alternatively, the peptide may comprise an alternative N-terminal group (i.e., N-terminal modification).

[0104] Thus, in some embodiments of the Kv1.3 inhibitors or pharmaceutically acceptable salts of the present invention, the peptide comprises at the N-terminus a group selected from 1-4 alkyl, acetyl (Ac), formyl, benzoyl, and trifluoroacetyl.

[0105] The “-OH” portion at the C-terminus of the sequence represents the presence of a carboxyl group (-COOH) at the C-terminus of the molecule. The “-NH2” portion at the C-terminus of the sequence represents the presence of an amide group (CONH2) at the C-terminus of the molecule. The “CH2OH” portion at the C-terminus represents the presence of a hydroxyl group linked to an alkyl group at the C-terminus of the molecule. The CH2OH portion may be included in (4-amino-5-hydroxypentyl)guanidine or 4-amino-5-hydroxypentanamide.

[0106] Thus, in some embodiments of the Kv1.3 inhibitors or pharmaceutically acceptable salts of the present invention, the peptide comprises at the C-terminus a carboxyl group (-COOH), an amino group (-NH2) or a hydroxymethyl group (-CH2OH), preferably a carboxyl group (-COOH) or an amino group (-NH2). Sequence

[0107] In the context of the Kv1.3 inhibitors of the present invention, the term "sequence" as used herein refers to the order of amino acids in the peptide of the Kv1.3 inhibitor. Sequence identifier numbers (SEQ ID NO) are used herein to refer to specific sequences. Each mention of a SEQ ID NO herein refers to the sequence represented by that SEQ ID NO. A peptide "comprising" a given sequence may contain additional amino acids at one or both ends of the sequence. A peptide "consisting of" a given sequence does not contain any amino acids other than those of the sequence in its linear sequence, but it may contain other features (such as chemical groups at the N-terminus or C-terminus).

[0108] In some embodiments of the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention, the peptide comprises or consists of one of the following sequences:

[0109] In some embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention comprises a peptide consisting of the sequence of any one of SEQ ID NOs 1 to 150. In some embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention consists of a peptide consisting of the sequence of any one of SEQ ID NOs 1 to 150. In some preferred embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention comprises a peptide consisting of the sequence of any one of SEQ ID NO 97. In some preferred embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention consists of a peptide consisting of the sequence of any one of SEQ ID NO 97. Peptide

[0110] In the context of the Kv1.3 inhibitors of the present invention, the term "peptide" as used herein refers to the peptide component of the Kv1.3 inhibitor. The term peptide encompasses additional features other than the peptide sequence (i.e., the order of amino acids), namely the chemical groups at the N-terminus and C-terminus of the peptide and the pattern of disulfide bridges in the peptide. For ease of reference, specific peptides are assigned peptide numbers (Ptd no.) herein. Each mention of a peptide number herein refers to the peptide represented by that peptide number. A Kv1.3 inhibitor "comprising" a given peptide may contain other features. A Kv1.3 inhibitor "consisting of" a given peptide contains only the features of that peptide.

[0111] In some embodiments of the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention, the peptide is selected from the following peptides:

[0112] In some embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention comprises a peptide consisting of any one of peptides 1 to 158. In some embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention consists of any one of peptides 1 to 158. In some preferred embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention comprises peptide 100. In some preferred embodiments, the Kv1.3 inhibitor or pharmaceutically acceptable salt of the present invention consists of peptide 100. Synthesis of the Kv1.3 inhibitor of the present invention

[0113] The Kv1.3 inhibitors described herein can be synthesized by solid-phase or liquid-phase peptide synthesis methods. In this case, reference can be made to WO 98 / 11125 and many other documents such as Fields, G.B. et al., 2002, “Principles and practice of solid-phase peptide synthesis”. In: Synthetic Peptides (Second Edition), as well as the examples herein. Alternatively, the Kv1.3 inhibitors described herein can be synthesized by recombinant techniques or by a combination of recombinant techniques and peptide chemistry.

[0114] Exemplary methods for producing the Kv1.3 inhibitor of the present invention include synthesizing a peptide by solid-phase or liquid-phase peptide synthesis methods and recovering the peptide; or expressing a peptide from a nucleic acid construct encoding the peptide and recovering the expression product; or expressing a precursor peptide from a nucleic acid construct encoding a precursor peptide sequence, recovering the expression product, and modifying the precursor peptide to produce a Kv1.3 inhibitor.

[0115] The precursor peptide can be modified by introducing one or more non-proteinogenic amino acids (such as Nle), introducing suitable terminal groups R 1 and R 2 and so on.

[0116] Expression of a peptide or precursor peptide from a nucleic acid encoding the peptide or precursor peptide can be carried out in a cell-free expression system or in a cell containing such a nucleic acid. Such expression generally requires that the peptide or precursor peptide be composed entirely of proteinogenic amino acids (i.e., the 20 amino acids encoded by the standard genetic code).

[0117] For recombinant expression, a nucleic acid fragment encoding a precursor peptide is typically inserted into a suitable vector to form a cloning or expression vector. Depending on the application purpose and type, the vector can be in the form of a plasmid, phage, cosmid, minichromosome, or virus, but naked DNA that is transiently expressed only in certain cells is also an important vector. Preferred cloning and expression vectors (plasmid vectors) are capable of autonomous replication, enabling the production of high copy numbers for high-level expression or high-level replication for subsequent cloning.

[0118] Generally, an expression vector contains the following features operably linked in the 5'→3' direction: a promoter for driving the expression of the nucleic acid fragment; optionally, a nucleic acid sequence encoding a leader peptide capable of being secreted (secreted into the extracellular phase or, where appropriate, into the periplasma); a nucleic acid fragment encoding the precursor peptide; and optionally, a nucleic acid sequence encoding a terminator. The expression vector may contain additional features, such as selectable markers and origins of replication. When manipulating the expression vector in a production strain or cell line, it is preferred that the vector be capable of integrating into the host cell genome. Those skilled in the art are very familiar with suitable vectors and can design vectors according to their specific requirements.

[0119] Such vectors are used to transform host cells to produce a peptide or precursor peptide. Such transformed cells can be cultured cells or cell lines used for amplifying the nucleic acid fragment and the vector and / or for recombinantly producing the precursor peptide.

[0120] Preferred transformed cells are microorganisms, such as bacteria (e.g., species of the genus Escherichia (e.g., Escherichia coli), Bacillus (e.g., Bacillus subtilis), Salmonella, or Mycobacterium (preferably non-pathogenic, such as Mycobacterium bovis BCG)), yeast (e.g., Saccharomyces cerevisiae and Pichia pastoris), and protozoa. Alternatively, the transformed cells can be derived from multicellular organisms, i.e., they can be fungal cells, insect cells, algal cells, plant cells, or animal cells (e.g., mammalian cells). For the purpose of cloning and / or optimizing expression, it is preferred that the transformed cells be capable of replicating the nucleic acid fragment of the present invention. Cells expressing the nucleic acid fragment can be used for small-scale or large-scale preparation of the peptide of the present invention. When producing a peptide or precursor peptide by transformed cells, it is convenient, although far from essential, to secrete the expression product into the culture medium. Treating or preventing a disease or disorder

[0121] The present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof as described herein for use in a method of treating or preventing a disease or disorder in a subject, wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

[0122] Accordingly, the present invention also provides a method of treating or preventing a disease or disorder in a subject, the method comprising administering a Kv1.3 inhibitor or a pharmaceutically acceptable salt of the present invention to the subject once every 2 to 8 days.

[0123] The present invention also provides the use of a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof as described herein in the manufacture of a medicament for use in a method of treating or preventing a disease or disorder in a subject, wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

[0124] In other words, the present invention provides the medical use of a Kv1.3 inhibitor or a pharmaceutically acceptable salt as described herein. The present invention provides therapeutic, prophylactic or preventive methods which comprise administering a Kv1.3 inhibitor or a pharmaceutically acceptable salt as described herein to a subject. In all cases, the medical use can also be expressed as a method of treating or preventing a disease or disorder, the method comprising administering a Kv1.3 inhibitor or a pharmaceutically acceptable salt of the present invention to a subject.

[0125] The term "subject" is used interchangeably herein with "patient" and "individual" and refers to a human or non-human animal. These terms include mammals such as humans, primates, domestic animals (e.g., cows and pigs), companion animals (e.g., dogs and cats) and rodents (e.g., mice and rats). The subject described herein suffers from the disease or disorder described herein.

[0126] The terms "disease" and "disorder" refer to a state of dysfunction of the body. The term disease is synonymous herein with similar terms such as "disorder" or "ailment". Accordingly, the terms "disease", "disorder" and "ailment" are interchangeable herein. In some embodiments, the disease or disorder is a disease or disorder that can be treated or prevented (i.e., is treatable or preventable) using a Kv1.3 inhibitor as described herein.

[0127] The term "treatment" and variations thereof refer to alleviating, reducing or eliminating the symptoms of a disease in a subject. Accordingly, the term "treatment" includes curing a disease or disorder, but does not require complete elimination of the disease or disorder from the subject. Even a minor alleviation of the symptoms of a disease or disorder is "treatment" of the disease or disorder. The term "treatment" and variations thereof refer to both treating an existing disease in a subject and preventing a disease (i.e., prophylaxis) in a subject. It will thus be appreciated that treatment as referred to herein can in some embodiments be prophylactic.

[0128] The terms "prevention", "prevent" and variations thereof relate to prophylactic treatment, i.e., to measures or steps that are aimed at preventing the occurrence of a disease or disorder rather than treating an existing disease or disorder. Prevention means obtaining such desired pharmacological and / or physiological effects that are prophylactic in completely or partially preventing a disease or disorder or its symptoms. Treating inflammation

[0129] As described herein, inhibitors of Kv1.3 can be used to reduce inflammation. The data presented in the examples herein directly demonstrate that treating a subject with a Kv1.3 inhibitor reduces inflammation.

[0130] Thus, in some embodiments, the disease or disorder is an inflammatory disease or disorder. An inflammatory disease or disorder is any disease, disorder or condition in which it is desirable to reduce inflammation, such as a condition in which inflammation contributes to the symptoms or pathogenesis.

[0131] In some embodiments, the inflammatory disorder or condition is selected from autoimmune diseases, allergy or hypersensitivity, allograft rejection, transplant rejection, graft-versus-host disease, hay fever, asthma, anaphylaxis, allergic rhinitis, urticaria, eczema, alopecia areata, dermatomyositis, inclusion body myositis, polymyositis, ankylosing spondylitis, vasculitis, arthritis (including rheumatoid arthritis, osteoarthritis, psoriatic arthritis), Sjogren's syndrome, systemic lupus erythematosus (SLE), uveitis, inflammatory fibrosis (such as scleroderma, pulmonary fibrosis, liver cirrhosis), chronic obstructive pulmonary disease (COPD), hepatitis, chronic inflammatory demyelinating polyneuropathy, inflammatory bowel disease, colitis (such as Crohn's disease and ulcerative colitis), erythema, thyroiditis, psoriasis, atopic dermatitis, allergic contact dermatitis, scleroderma, glomerulonephritis, inflammatory bone resorption, multiple sclerosis and type 1 diabetes. In some preferred embodiments, the disease or disorder is arthritis, such as rheumatoid arthritis, osteoarthritis or psoriatic arthritis.

[0132] In some embodiments, the present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof of the present invention as described herein, for use in a method of inhibiting or reducing inflammation.

[0133] In some embodiments, after administration of a Kv1.3 inhibitor, inflammation in the subject is reduced. Inflammation can be measured using techniques known in the art (e.g., by measuring cytokine levels in the subject). In some embodiments, after administration of a Kv1.3 inhibitor, inflammation in the subject is reduced by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95%. In some embodiments, after administration of a Kv1.3 inhibitor, inflammation in the subject is reduced by 100% (i.e., inflammation is eliminated). Treatment of metabolic diseases

[0134] Inhibitors of Kv1.3 can also have beneficial metabolic effects, such as those related to energy homeostasis, body weight regulation, and glucose control.

[0135] In some embodiments, the disease or disorder is a metabolic disease or disorder. A metabolic disease or disorder is any disease, disorder, or condition caused by or associated with abnormal metabolism (i.e., any disruption of the body's processing of food to sustain life). A metabolic disease or disorder can be characterized by the presence of too much or too little of a particular chemical substance in the body of the subject, such as proteins, carbohydrates, lipids, their constituent molecules (e.g., amino acids, sugars, fatty acids), and other such biomolecules.

[0136] Given that obese subjects have too much fat and can have abnormal metabolic processes, obesity can be considered a metabolic disorder. Thus, in some embodiments, the disease or disorder is obesity, obesity-related inflammation, obesity-related gallbladder disease, or obesity-induced sleep apnea.

[0137] In some embodiments, the disease or disorder is a disease or disorder caused by or associated with impaired glucose control. Such diseases include metabolic syndrome, insulin resistance, glucose intolerance, prediabetes, elevated fasting blood glucose, and type 2 diabetes. Some of these conditions may be related to obesity. The effects of Kv1.3 inhibitors on these conditions may be mediated, in whole or in part, by their effects on body weight, or may be independent of their effects on body weight.

[0138] In some embodiments, the present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt of the present invention as described herein for use in a method of inhibiting weight gain, promoting weight loss, reducing overweight body weight, or treating obesity (e.g., by controlling appetite, feeding, food intake, calorie intake, and / or energy expenditure). The effect on body weight can be therapeutic or cosmetic. Treatment of proliferating cells and cancer

[0139] Kv1.3 is also expressed in proliferating human and mouse smooth muscle cells. Inhibitors of Kv1.3 can be effective against smooth muscle proliferative disorders such as restenosis (e.g., in patients who have undergone vascular surgery (e.g., angioplasty)). Thus, in some embodiments, the disease or disorder is a smooth muscle proliferative disorder. In some embodiments, the smooth muscle proliferative disorder is restenosis.

[0140] Additional evidence indicates that the Kv1.3 channel is involved in the activation and / or proliferation of multiple cell types, including tumor cells (Bielanska et al., Curr. Cancer Drug Targets 9:904-14, 2009), microglia (Khanna et al., Am. J. Physiol. Cell Physiol. 280:C796-806, 2001), and the differentiation of neuronal progenitor cells (Wang et al., J. Neurosci. 30:5020-7, 2010). Thus, Kv1.3 inhibitors may be beneficial for treating neuroinflammatory and neurodegenerative disorders. Thus, in some embodiments, the disease or disorder is a neuroinflammatory or neurodegenerative disease or disorder. In some embodiments, the neuroinflammatory or neurodegenerative disease or disorder is Alzheimer’s disease, multiple sclerosis (MS), Parkinson’s disease, or amyotrophic lateral sclerosis (ALS) (e.g., after viral infection).

[0141] In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is breast cancer, prostate cancer, or lymphoma. In some embodiments, the lymphoma is non-Hodgkin lymphoma (NHL). Non-Hodgkin lymphoma includes T-cell NHL and B-cell NHL. Forms of B-cell NHL include diffuse large B-cell lymphoma, follicular lymphoma, Burkitt lymphoma, immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma, and mantle cell lymphoma. Forms of T-cell NHL include mycosis fungoides, anaplastic large cell lymphoma, peripheral T-cell lymphoma, precursor T lymphoblastic lymphoma, and Sézary syndrome. Administration of a Kv1.3 inhibitor

[0142] The present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use in a method of treating or preventing a disease or disorder in a subject, wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days. Thus, the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days. In other words, the Kv1.3 inhibitor is administered to the subject once every 2 to 8 days.

[0143] The term "administer" and variations thereof refer to providing the Kv1.3 inhibitor to the interior of a subject. "One administration" is a single event of administering the Kv1.3 inhibitor to a subject. Administration of the Kv1.3 inhibitor can be by any mode of administration common or standard in the art, such as oral, intravenous, intramuscular, subcutaneous, sublingual, intranasal, or intradermal administration, by the rectal route or by implantation. In some embodiments, the Kv1.3 inhibitor is administered by injection, preferably subcutaneous injection. Time of administration

[0144] According to the present invention, the Kv1.3 inhibitor is administered to the subject once every 2 to 8 days.

[0145] In other words, according to the present invention, the Kv1.3 inhibitor is administered to the subject once every 2 to 8 days at a specific dose. In some aspects of the present invention, the Kv1.3 inhibitor is administered to the subject once every 3 to 8 days. In other words, in some aspects of the present invention, the Kv1.3 inhibitor is administered to the subject once every 3 to 8 days at a specific dose. In some aspects of the present invention, the Kv1.3 inhibitor is administered to the subject once every 3 to 7 days. In other words, in some aspects of the present invention, the Kv1.3 inhibitor is administered to the subject once every 3 to 7 days at a specific dose.

[0146] Kv1.3 inhibitors based on the PaT1 toxin peptide have a short half-life in the body of a subject. For example, Example 6 herein shows that such a Kv1.3 inhibitor has a half-life of about 1 hour in a rat model organism. However, it was unexpectedly found that despite this short half-life, the Kv1.3 inhibitors described herein also have a lasting effect on T cells. In particular, Example 8 herein shows that inflammation in a rat ear inflammation model was reduced for up to 7 days after treatment with a Kv1.3 inhibitor. Additionally, Example 9 herein shows that treatment with a Kv1.3 inhibitor once every 5 days reduced inflammation in a rat arthritis model. Without wishing to be bound by theory, it is hypothesized that the Kv1.3 inhibitor somehow "reprograms" T cells through its interaction with the Kv1.3 ion channel to remain inactive for an extended period of time, thereby suppressing inflammation for multiple days.

[0147] These unexpected findings support the concept of treating a subject having a disease, disorder or condition treatable with a Kv1.3 inhibitor as described herein according to a regimen of administering the inhibitor at approximately weekly intervals (i.e., once every 2 to 8 days). It is advantageous to extend the time period between administrations of the active compound as much as possible because this saves time and effort (since fewer administrations are required over a given time period) and cost (since less of the active compound is required over a given time period). In particular, in the case of administering the active compound by certain routes (such as subcutaneous injection), longer intervals between administrations also improve patient comfort and can thereby improve patient compliance with the treatment regimen. For example, for the reasons described above, administering a Kv1.3 inhibitor according to the present invention (i.e., once every 2 to 8 days) is superior to daily administration of the inhibitor. The unexpected finding described in the present disclosure is that it is possible to administer a Kv1.3 inhibitor as described herein once every 2 to 8 days despite its short in vivo half-life because an effect of the inhibitor (such as reduced inflammation) was observed for multiple days after administration.

[0148] Accordingly, a method of treating a disease or disorder comprises administering a Kv1.3 inhibitor to a subject once every 2 to 8 days. In other words, the Kv1.3 inhibitor is administered to the subject at least 2 days after a previous administration of the Kv1.3 inhibitor (if any) and at most 8 days after a previous administration of the Kv1.3 inhibitor to the subject. The time period between administrations of the Kv1.3 inhibitor to the subject may be referred to as the "interval" between administrations of the Kv1.3 inhibitor. Accordingly, the method comprises administering a Kv1.3 inhibitor to the subject at an interval of 2 to 8 days (i.e., the interval between administrations is a duration of 2 to 8 days). The method comprises administering a Kv1.3 inhibitor to the subject at an interval of 2 to 8 days.

[0149] The term "day" refers to a time period of 24 hours ± 8 hours (i.e., 16 to 32 hours). In other words, "1 day" is approximately 24 hours, allowing for an 8-hour margin of error around the exact 24-hour time point. The 8-hour margin of error is not cumulative over multiple days. Thus, "2 days" refers to a time period of 48 hours ± 8 hours (i.e., 40 to 56 hours) rather than a time period of 48 hours ± 16 hours, "3 days" refers to a time period of 72 hours ± 8 hours (i.e., 64 to 80 hours), "4 days" refers to a time period of 96 hours ± 8 hours (i.e., 88 to 104 hours), "5 days" refers to a time period of 120 hours ± 8 hours (i.e., 112 to 128 hours), "6 days" refers to a time period of 144 hours ± 8 hours (i.e., 136 to 152 hours), "7 days" refers to a time period of 168 hours ± 8 hours (i.e., 160 to 176 hours) and "8 days" refers to a time period of 192 hours ± 8 hours (i.e., 184 to 200 hours).

[0150] Thus, the expression "administering a Kv1.3 inhibitor to a subject once every 2 to 8 days" (i.e., once every 2 days to 8 days) herein means administering a Kv1.3 inhibitor to a subject once every 40 to 200 hours. Thus, the Kv1.3 inhibitor is administered to the subject 40 hours to 200 hours after a previous administration (if any). In other words, the Kv1.3 inhibitor is administered to the subject at least 40 hours after a previous administration (if any) of the Kv1.3 inhibitor to the subject and at most 200 hours after a previous administration (if any) of the Kv1.3 inhibitor to the subject.

[0151] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 2 to 7 days, such as once every 2 to 6 days, once every 2 to 5 days, once every 2 to 4 days, or once every 2 to 3 days. In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 3 to 8 days, such as once every 3 to 7 days, once every 3 to 6 days, once every 3 to 5 days, or once every 3 to 4 days. In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 4 to 8 days, such as once every 4 to 7 days, once every 4 to 6 days, or once every 4 to 5 days. In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 5 to 8 days, such as once every 5 to 7 days or once every 5 to 6 days. In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 6 to 8 days, such as once every 6 to 7 days. In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 7 to 8 days.

[0152] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, or once every 8 days. Preferably, the method comprises administering a Kv1.3 inhibitor to a subject once every 7 days. Preferably, the method comprises administering a Kv1.3 inhibitor to a subject approximately once a week. Preferably, the method comprises administering a Kv1.3 inhibitor to a subject once a week.

[0153] The method comprises administering a Kv1.3 inhibitor to a subject once every 2 to 8 days within an administration period. "Administration period" is the total time period during which a Kv1.3 inhibitor is administered to a subject (i.e., the time period during which the Kv1.3 is administered to the subject at intervals). In other words, the administration period is the time period starting from the first administration of the Kv1.3 inhibitor during that administration period to the end of the last administration of the inhibitor during that administration period. The duration of the administration period can depend on various factors, including whether a disease or disorder is being treated or prevented, the type of disease or disorder being treated or prevented, and the characteristics of the subject (such as age, weight, or immune status). The administration period can be determined by a clinician prescribing the Kv1.3 inhibitor for the subject.

[0154] The administration period can be as long as required for treating or preventing a disease or disorder in a subject (i.e., the administration of the Kv1.3 inhibitor can continue for as long as required). In some embodiments, the administration period is at least 1 month. In other words, in some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days for at least 1 month. In some embodiments, the administration period is at least 2 months, such as at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months (i.e., at least 1 year). In some embodiments, the administration period is at least 1 year. In other words, in some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days for at least 1 year. In some embodiments, the administration period is at least 2 years, such as at least 3 years, at least 4 years, at least 5 years, at least 6 years, at least 7 years, at least 8 years, at least 9 years, at least 10 years, at least 11 years, at least 12 years, at least 13 years, at least 14 years, at least 15 years, at least 16 years, at least 17 years, at least 18 years, at least 19 years, or at least 20 years. In some embodiments, the administration period is for the lifetime of the subject. In other words, in some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days for the lifetime of the subject.

[0155] A given subject can undergo multiple (i.e., more than one) administration periods. In other words, the Kv1.3 can be administered to the subject during a given administration period, then the administration of the Kv1.3 inhibitor can be stopped, and then the Kv1.3 inhibitor can be administered to the subject again for another administration period. There can be multiple such interruptions and resumptions of the administration of the Kv1.3 inhibitor during the lifetime of the subject (i.e., multiple administration periods, such as 2, 3, 4, 5, 6, 7, 8, 9, 10 or more administration periods). The administration period of the present invention can be any one (or more) of these multiple administration periods and is not excluded by the fact that one or more other administration periods are not within the scope of the present invention.

[0156] The number of Kv1.3 inhibitor administrations to a subject during a dosing period (i.e., the number of times the Kv1.3 inhibitor is administered to the subject during a dosing period) depends on the duration of the dosing period and the time elapsed between administrations (i.e., the interval between administrations). For example, the longer the dosing period, the greater the total number of times the Kv1.3 inhibitor may be administered to the subject during that dosing period. The Kv1.3 inhibitor may be administered to the subject any number of times (i.e., the method includes administering the Kv1.3 inhibitor to the subject any number of times). In some embodiments, the method includes administering the Kv1.3 inhibitor to the subject at least 2 times (i.e., 2 or more times) during the dosing period. In other words, in some embodiments, the method includes administering the Kv1.3 inhibitor to the subject at least 2 times during the dosing period. In some embodiments, the method includes administering the Kv1.3 inhibitor to the subject at least 3 times (i.e., 3 or more times) during the dosing period, such as at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, or at least 100 times during the dosing period.

[0157] Generally, the interval between Kv1.3 inhibitor administrations (i.e., the amount of time elapsed from a given administration until the next administration) is the same throughout the dosing period. In other words, in some embodiments, the interval between Kv1.3 inhibitor administrations is constant or uniform throughout the dosing period. For example, the Kv1.3 inhibitor may be administered once every 7 days (i.e., the interval between all administrations is 7 days). A constant interval is preferred because it can improve patient compliance with the treatment, as it is simpler to administer the inhibitor at a constant interval (e.g., it may be easier for the subject / patient to remember to administer the inhibitor). However, in some embodiments, the intervals are different. In other words, in some embodiments, each interval is independently selected to be a duration of 2 to 8 days. For example, the Kv1.3 inhibitor may be administered 2 days after a previous administration, which may have been 8 days after its previous administration, which may have been 5 days after its previous administration, and so on. Under the guidance of a clinician, the intervals may be varied in this way to seek to achieve a particular clinical outcome. For example, the intervals may be increased during the course of the dosing period to determine the maximum possible interval length while maintaining treatment or preventing a disease or disorder.

[0158] According to the present invention, the method comprises administering a Kv1.3 inhibitor to a subject once every 2 to 8 days. However, within a given administration period, there may be intervals lasting longer than 8 days. It should be understood that the present invention encompasses administering a Kv1.3 inhibitor once every 2 to 8 days for any part of an administration period. For example, administering a Kv1.3 inhibitor to a subject once every 2 to 8 days for an initial part of the administration period, and subsequently administering a Kv1.3 inhibitor to the subject at intervals longer than 8 days is within the scope of the present invention. Thus, according to the present invention, a method of treating or preventing a disease or disorder comprises administering a Kv1.3 inhibitor to a subject at least twice, wherein the interval between at least two administrations of the Kv1.3 inhibitor is 2 to 8 days. In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject, wherein there is an interval of 2 to 8 days between at least two administrations of the Kv1.3 inhibitor. In some embodiments, the method comprises at least one interval of 2 to 8 days (i.e., the interval between administrations of the Kv1.3 inhibitor). In other words, in some embodiments, the method comprises a first administration of a Kv1.3 inhibitor to a subject, followed by a second administration of the Kv1.3 inhibitor 2 to 8 days later. The Kv1.3 inhibitor may be additionally administered to the subject before or after the first and second administrations. In some embodiments, the method consists of a first administration of a Kv1.3 inhibitor to a subject followed by a second administration of the Kv1.3 inhibitor 2 to 8 days later (i.e., there is no additional administration of the Kv1.3 inhibitor). Dosage of the Kv1.3 inhibitor

[0159] The term "dose" refers to the amount of the Kv1.3 inhibitor administered to a subject in a single administration. For example, in some embodiments, where the Kv1.3 inhibitor is administered by subcutaneous injection, the "dose" may refer to the amount of the Kv1.3 inhibitor in a single injection of the inhibitor. The terms "dosage" and "dose" may be used interchangeably herein.

[0160] In some embodiments, the method comprises administering the Kv1.3 inhibitor in increasing doses.

[0161] In some embodiments, the method comprises administering the Kv1.3 inhibitor to a subject at a dose of 0.1 mg to 30.0 mg.

[0162] In other words, in some embodiments, the Kv1.3 inhibitor is administered to a subject at a dose of 0.1 mg to 30.0 mg. In other words, in some embodiments, the Kv1.3 inhibitor is administered to a subject at a dose of not less than 0.1 mg and not more than 30.0 mg. In some embodiments, the Kv1.3 inhibitor is formulated at a dose of 0.1 mg to 30.0 mg. In some embodiments, the method comprises administering the Kv1.3 inhibitor to a subject once every 2 to 8 days at a dose of 0.1 mg to 30.0 mg.

[0163] In some embodiments, the dose of the Kv1.3 inhibitor administered to the subject each time is the same as the doses administered in other instances. In some embodiments, the dose of the Kv1.3 inhibitor administered to the subject each time can be different from the doses administered in other instances. In other words, the dose of the Kv1.3 inhibitor administered to the subject each time can be independently selected to be from 0.1 mg to 30.0 mg.

[0164] In some embodiments, the Kv1.3 inhibitor is administered to the subject in a single-dose formulation of from 0.1 mg to 30.0 mg. The single-dose formulation can be administered to the subject one or more times, and each dose in the multi-dose formulation used for administering to the subject does not need to contain the same amount of the Kv1.3 inhibitor. In other words, the Kv1.3 inhibitor can be administered to the subject in a series of single administrations, and each of the single administrations may not contain the same amount of the Kv1.3 inhibitor.

[0165] In some embodiments, the method comprises administering a Kv1.3 inhibitor to the subject at a dose of: from 1.0 mg to 30.0 mg, such as from 2.0 mg to 30.0 mg, from 3.0 mg to 30.0 mg, from 4.0 mg to 30.0 mg, from 5.0 mg to 30.0 mg, from 6.0 mg to 30.0 mg, from 7.0 mg to 30.0 mg, from 8.0 mg to 30.0 mg, from 9.0 mg to 30.0 mg, from 10.0 mg to 30.0 mg, from 11.0 mg to 30.0 mg, from 12.0 mg to 30.0 mg, from 13.0 mg to 30.0 mg, from 14.0 mg to 30.0 mg, from 15.0 mg to 30.0 mg, from 16.0 mg to 30.0 mg, from 17.0 mg to 30.0 mg, from 18.0 mg to 30.0 mg, from 19.0 mg to 30.0 mg, from 20.0 mg to 30.0 mg, from 21.0 mg to 30.0 mg, from 22.0 mg to 30.0 mg, from 23.0 mg to 30.0 mg, from 24.0 mg to 30.0 mg, from 25.0 mg to 30.0 mg, from 26.0 mg to 30.0 mg, from 27.0 mg to 30.0 mg, from 28.0 mg to 30.0 mg, or from 29.0 mg to 30.0 mg.

[0166] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of from 1.0 mg to 29.0 mg, such as from 1.0 mg to 28.0 mg, from 1.0 mg to 27.0 mg, from 1.0 mg to 26.0 mg, from 1.0 mg to 25.0 mg, from 1.0 mg to 24.0 mg, from 1.0 mg to 23.0 mg, from 1.0 mg to 22.0 mg, from 1.0 mg to 21.0 mg, from 1.0 mg to 20.0 mg, from 1.0 mg to 19.0 mg, from 1.0 mg to 18.0 mg, from 1.0 mg to 17.0 mg, from 1.0 mg to 16.0 mg, from 1.0 mg to 15.0 mg, from 1.0 mg to 14.0 mg, from 1.0 mg to 13.0 mg, from 1.0 mg to 12.0 mg, from 1.0 mg to 11.0 mg, from 1.0 mg to 10.0 mg, from 1.0 mg to 9.0 mg, from 1.0 mg to 8.0 mg, from 1.0 mg to 7.0 mg, from 1.0 mg to 6.0 mg, from 1.0 mg to 5.0 mg, from 1.0 mg to 4.0 mg, from 1.0 mg to 3.0 mg, or from 1.0 mg to 2.0 mg.

[0167] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of from 0.1 mg to 15.0 mg, such as from 0.1 mg to 10.0 mg, from 0.1 mg to 9.0 mg, from 0.1 mg to 8.0 mg, from 0.1 mg to 7.0 mg, from 0.1 mg to 6.0 mg, from 0.1 mg to 5.0 mg, from 0.1 mg to 4.0 mg, from 0.1 mg to 3.0 mg, from 0.1 mg to 2.0 mg, from 0.1 mg to 1.0 mg, or from 0.1 mg to 0.5 mg.

[0168] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of about 1.0 mg, about 2.0 mg, about 3.0 mg, about 4.0 mg, about 5.0 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10.0 mg, about 11.0 mg, about 12.0 mg, about 13.0 mg, about 14.0 mg, about 15.0 mg, about 16.0 mg, about 17.0 mg, about 18.0 mg, about 19.0 mg, about 20.0 mg, about 21.0 mg, about 22.0 mg, about 23.0 mg, about 24.0 mg, about 25.0 mg, about 26.0 mg, about 27.0 mg, about 28.0 mg, about 29.0 mg, or about 30.0 mg.

[0169] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of from 10 nmol / kg to 400 nmol / kg.

[0170] In other words, in some embodiments, the Kv1.3 inhibitor is administered to a subject at a dose of 10 nmol / kg to 400 nmol / kg. In other words, in some embodiments, the Kv1.3 inhibitor is administered to a subject at a dose of no less than 10 nmol / kg and no more than 400 nmol / kg. In some embodiments, the Kv1.3 inhibitor is formulated at a dose of 10 nmol / kg to 400 nmol / kg. In some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject at a dose of 10 nmol / kg to 400 nmol / kg once every 2 to 8 days. In some embodiments, the dose of the Kv1.3 inhibitor administered to the subject each time can be independently selected to be 10 nmol / kg to 400 nmol / kg.

[0171] The unit of "nmol / kg" means that the dose of the Kv1.3 inhibitor is related to the weight of the subject per unit weight. A given number of nanomoles of Kv1.3 is administered per kilogram of the subject's body weight. Therefore, the exact dose of the Kv1.3 inhibitor can be determined by the clinician on a case-by-case basis.

[0172] In some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject at the following doses: 50 nmol / kg to 400 nmol / kg, such as 100 nmol / kg mg to 400 nmol / kg, 150 nmol / kg mg to 400 nmol / kg, 200 nmol / kgmg to 400 nmol / kg, 250 nmol / kg mg to 400 nmol / kg, 300 nmol / kg mg to 400 nmol / kg, or 350 nmol / kg mg to 400 nmol / kg.

[0173] In some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject at the following doses: 10 nmol / kg to 350 nmol / kg, such as 10 nmol / kg mg to 350 nmol / kg, 10 nmol / kg mg to 300 nmol / kg, 10 nmol / kg mg to 250 nmol / kg, 10 nmol / kg mg to 200 nmol / kg, 10 nmol / kg mg to 150 nmol / kg, 10 nmol / kg mg to 100 nmol / kg, or 10 nmol / kg mg to 50 nmol / kg.

[0174] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of about 3 nmol / kg, about 10 nmol / kg, about 50 nmol / kg, about 100 nmol / kg, about 150 nmol / kg, about 200 nmol / kg, about 250 nmol / kg, about 300 nmol / kg, about 350 nmol / kg, or about 400 nmol / kg. Preferably, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of about 300 nmol / kg.

[0175] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of from about 3 nmol / kg to about 300 nmol / kg.

[0176] In other words, in some embodiments, the Kv1.3 inhibitor is administered to the subject at a dose of from 3 nmol / kg to 300 nmol / kg. In some embodiments, the Kv1.3 inhibitor is administered to the subject at a dose of no less than 3 nmol / kg and no more than 300 nmol / kg. In some embodiments, the Kv1.3 inhibitor is formulated at a dose of from 3 nmol / kg to 300 nmol / kg. In some embodiments, the method comprises administering the Kv1.3 inhibitor to the subject at a dose of from 3 nmol / kg to 300 nmol / kg once every 2 to 8 days. In some embodiments, the dose of the Kv1.3 inhibitor administered to the subject each time can be independently selected to be from 3 nmol / kg to 300 nmol / kg.

[0177] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of: 50 nmol / kg to 300 nmol / kg, such as 100 nmol / kg mg to 300 nmol / kg, 150 nmol / kg mg to 300 nmol / kg, 200 nmol / kgmg to 300 nmol / kg, or 250 nmol / kg mg to 300 nmol / kg.

[0178] In some embodiments, the method comprises administering a Kv1.3 inhibitor to a subject at a dose of: 3 nmol / kg to 250 nmol / kg, such as 3 nmol / kg mg to 200 nmol / kg, 3 nmol / kg mg to 150 nmol / kg, 3 nmol / kg mg to 100 nmol / kg, or 3 nmol / kg mg to 50 nmol / kg. Pharmaceutical composition

[0179] The present invention also provides a pharmaceutical composition comprising a Kv1.3 inhibitor or a pharmaceutically acceptable salt as described herein, for use in the methods as described herein. Accordingly, the present invention provides a pharmaceutical composition comprising a Kv1.3 inhibitor or a pharmaceutically acceptable salt as described herein, for use in a method of treating or preventing a disease or disorder in a subject, wherein the method comprises administering the pharmaceutical composition to the subject once every 2 to 8 days.

[0180] In other words, in some embodiments, the present invention provides a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof, for use as described herein, wherein the Kv1.3 inhibitor or pharmaceutically acceptable salt is in the form of a composition. Preferably, the composition is a pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable carrier, excipient or vehicle. In some embodiments, the pharmaceutical composition of the present invention is a pharmaceutical composition wherein the Kv1.3 inhibitor is in the form of a pharmaceutically acceptable acid addition salt.

[0181] In some embodiments, the composition comprises one or more Kv1.3 inhibitors as described herein (i.e., more than one Kv1.3 inhibitor). Each of the Kv1.3 inhibitors is independently selected from any of the Kv1.3 inhibitors described herein. In other words, each of the Kv1.3 inhibitors in the composition can be any of the Kv1.3 inhibitors described herein. In some embodiments, the composition comprises one or more peptides. In some embodiments, the composition comprises one or more peptides, wherein each of the peptides comprises a sequence independently selected from any of the peptide sequences described herein or consists of a sequence independently selected from any of the peptide sequences described herein. In some embodiments, the composition comprises one or more peptides, wherein each of the peptides comprises a peptide independently selected from any of the peptides described herein or consists of a peptide independently selected from any of the peptides described herein.

[0182] In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof. As will be apparent to those skilled in the medical arts, the "therapeutically effective amount" of the compounds or pharmaceutical compositions of the present invention will vary particularly depending on the age, weight and / or sex of the subject (patient) to be treated. Other factors that may be relevant include the physical characteristics of the particular patient under consideration, the patient's diet, the nature of any concomitant medications, the particular compound used, the particular mode of administration, the desired pharmacological effect and the particular therapeutic indication. Since these factors and their relationship in determining a therapeutically effective amount are well known in the medical arts, determining a therapeutically effective dosage level for achieving the desired therapeutic effect will be within the capabilities of the skilled person.

[0183] As used herein, the term "therapeutically effective amount" means an amount that alleviates the symptoms of a given disease, disorder, condition or pathological state and preferably normalizes the physiological response in an individual suffering from the disease, disorder, condition or pathological state. The alleviation of symptoms or the normalization of physiological response can be determined using conventional methods in the art and may vary with the given disease, disorder, condition or pathological state. In one aspect, a therapeutically effective amount of a Kv1.3 inhibitor or pharmaceutical composition of the invention is an amount that restores a measurable physiological parameter to a value substantially the same as that in an individual not suffering from the disease, disorder, condition or pathological state under discussion.

[0184] In some embodiments of the invention, administration of the Kv1.3 inhibitor or pharmaceutical composition of the invention is initiated at a lower dose level and the dose level is increased until the desired effect for the prophylactic / therapeutic relevant medical indication is achieved. This will define the therapeutically effective amount. For the compounds of the invention (alone or as part of a pharmaceutical composition), such a human dose of the active compound can be from about 0.01 pmol / kg to 500 μmol / kg body weight, from about 0.01 pmol / kg to about 300 μmol / kg body weight, from about 0.01 pmol / kg to about 100 μmol / kg body weight, from about 0.1 pmol / kg to about 50 μmol / kg body weight, from about 1 pmol / kg to about 10 μmol / kg body weight, from about 5 pmol / kg to about 5 μmol / kg body weight, from about 10 pmol / kg to about 1 μmol / kg body weight, from about 50 pmol / kg to about 0.1 μmol / kg body weight, from about 100 pmol / kg to about 0.01 μmol / kg body weight, from about 0.001 μmol / kg to about 0.5 μmol / kg body weight, from about 0.05 μmol / kg to about 0.1 μmol / kg body weight.

[0185] Effective doses and treatment regimens can be determined by conventional means, starting at low doses in laboratory animals and then increasing the dose while monitoring the effect, and also systematically varying the dosing regimen. When determining the optimal dose for a given subject, the clinician may consider several factors. Such considerations are known to those skilled in the art. Patent terms

[0186] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings commonly understood by those of ordinary skill in the art. Generally speaking, the nomenclature and techniques associated with chemistry, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry as described herein are those well known and commonly used in the art.

[0187] All patents, published patent applications, and non-patent publications mentioned in this application are hereby specifically incorporated by reference. In case of conflict, the present specification (including its specific definitions) shall prevail.

[0188] Each embodiment of the invention described herein can be used alone or in combination with one or more other embodiments of the invention.

[0189] The present disclosure is not limited to the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the embodiments of the present disclosure. Numerical ranges include the numbers defining the range. Unless otherwise specified, any nucleic acid sequence is written from left to right in the 5' to 3' direction; amino acid sequences are written from left to right in the amino to carboxyl direction.

[0190] Unless the context clearly dictates otherwise, a noun without a quantifier means one or more.

[0191] Throughout this specification, the word "comprise" and its grammatical variants (e.g., "comprises" or "comprising") shall be understood to mean including the specified whole or component or group of wholes or components, but not excluding any other whole or component or group of wholes or components. The terms "comprise" and "consist of" used herein are synonymous with "include" or "contain", and are inclusive or open-ended, without excluding additional unrecited members, elements or method steps. The terms "comprise" and "consist of" also include the term "consist of". The term "include" is used to mean "including but not limited to". "Include" and "including but not limited to" can be used interchangeably.

[0192] The publications discussed herein are provided only for their disclosure prior to the filing date of this application. Nothing herein is to be construed as an admission that such publications constitute prior art to the aspects appended hereto.

[0193] The present invention will now be further described by way of examples, which are intended to assist those of ordinary skill in the art in practicing the present invention and are not intended to limit the scope of the present invention in any way. Sequence Listing

[0194] The following sequence identifier numbers (SEQ ID NO) are assigned to the sequences herein: SEQ ID NO: 1 to 150 - Kv1.3 inhibitor peptide sequences SEQ ID NO: 151 to 180 & 182 to 188 - Fragments of Kv1.3 inhibitor peptide sequences SEQ ID NO: 181 - Skipped reading Example Example 1: General peptide synthesis

[0195] Table 1 below provides a list of abbreviations and suppliers. Table 1 Apparatus and synthesis strategy

[0196] According to solid-phase peptide synthesis procedures, peptides were synthesized batchwise on a peptide synthesizer (such as a CEM Liberty peptide synthesizer or a Symphony X synthesizer) using 9-fluorenylmethyloxycarbonyl (Fmoc) as the N-α-amino protecting group and appropriate common protecting groups for side-chain functional groups.

[0197] A polymer support-based resin such as TentaGel was used TM . The resin swollen in DMF before use was loaded into the synthesizer. Coupling CEM Liberty peptide synthesizer

[0198] A solution of Fmoc-protected amino acid (4 equivalents) was added to the resin together with a solution of coupling reagent (4 equivalents) and a base solution (8 equivalents). The mixture was heated to 70 to 75 °C in a microwave unit and coupled for 5 minutes or coupled for 60 minutes without heating. During coupling, nitrogen was bubbled through the mixture. Symphony X synthesizer

[0199] The coupling solution was transferred to the reaction vessel in the following order: amino acid (4 equivalents), HATU (4 equivalents), and DIPEA (8 equivalents). Unless otherwise specified, the coupling time was 10 minutes at room temperature (RT). The resin was washed with DMF (5 × 0.5 minutes). In the case of repeated coupling, the coupling time was 45 minutes at room temperature in all cases. Deprotection CEM Liberty peptide synthesizer

[0200] Deprotect the Fmoc group using piperidine in DMF or other suitable solvents. Add the deprotection solution to the reaction vessel and heat the mixture for 30 seconds to reach approximately 40 °C. Empty the reaction vessel, add fresh deprotection solution, and then heat to 70 to 75 °C for 3 minutes. After emptying the reaction vessel, wash the resin with DMF or other suitable solvents. Symphony X synthesizer

[0201] Perform Fmoc deprotection using 40% piperidine in DMF for 2.5 minutes and repeat using the same conditions. Wash the resin with DMF (5 × 0.5 minutes). Cleavage

[0202] Treat the dried peptide resin with TFA and suitable scavengers for approximately 2 hours. Reduce the volume of the filtrate and precipitate the crude peptide after adding diethyl ether. Wash the crude peptide precipitate with diethyl ether several times and finally dry it. HPLC purification of the crude peptide

[0203] Purify the crude peptide by preparative reversed-phase HPLC: Use a conventional HPLC apparatus, such as a Gilson GX-281, with a 331 / 332 pump combination for binary gradient applications, equipped with a column (e.g., a 5 × 25 cm Gemini NX 5u C18 110A column) and a fraction collector, using a flow rate of 20 to 40 ml / min with an appropriate gradient of buffer A (0.1% formic acid, aqueous solution) or A (0.1% TFA, aqueous solution) and buffer B (0.1% formic acid, 90% MeCN, aqueous solution) or B (0.1% TFA, 90% MeCN, aqueous solution). Analyze the fractions by analytical HPLC and MS, combine the selected fractions, and lyophilize. Characterize the final product by HPLC and MS. Formation of disulfide bonds

[0204] Dissolve the crude or partially purified linear peptide with six cysteines in a buffer (e.g., sodium bicarbonate (NaHCO3) or ammonium acetate (NH4Ac)) to obtain a final concentration of approximately 0.1 mg / ml or 25 μM. Adjust the pH of the buffer to pH 8.0 and stir the solution with magnetic stirring at room temperature and open the channel to the atmosphere. The reaction progress is determined by HPLC and is generally evaluated as completed overnight. Quench the solution by lowering the pH of the solution with an organic acid (e.g., acetic acid or trifluoroacetic acid (pH < 4)). Filter the solution and load it directly onto a preparative HPLC column for purification. Analytical HPLC

[0205] The final purity was determined by analytical HPLC (Agilent 1100 / 1200 series) equipped with an autosampler, a degasser, a 20 μl flow cell, and Chromeleon software. The HPLC was operated at a flow rate of 1.2 ml / min at 40 °C using an analytical column (e.g., Kinetex 2.6 μm XB-C18 100A 100×4.6 mm column). The compounds were detected and quantified at 215 nm. Buffer A (0.1% TFA, aqueous solution) and Buffer B (0.1% TFA, 90% MeCN, aqueous solution). Mass spectrometry

[0206] The final MS analysis was performed on a conventional mass spectrometer (e.g., Waters Xevo G2 Tof) equipped with an electrospray detector with lock mass calibration and MassLynx software. It was operated in positive mode using direct injection and a cone voltage of 15 V (1TOF), 30 V (2TOF), or 45 V (3TOF) as specified in the chromatogram. The accuracy was 5 ppm, while the typical resolution was 15,000 to 20,000.

[0207] The synthetic peptides (i.e., Kv1.3 inhibitors) are shown in Table 2: Table 2 Example 2: Selectivity of Kv1.3 Inhibitor Peptides in Patch-Clamp Assays

[0208] Patch-clamp assays were used to determine the selectivity of Kv1.3 inhibitor peptides for Kv1.3 over other potassium channels (Kv1.1, Kv1.2, and Kv1.6).

[0209] Chinese Hamster Ovary (CHO) cell lines stably expressing exogenous human α-subunits of each potassium channel were grown and passaged under standard culture conditions.

[0210] An automated chip-based planar patch-clamp device was used Quantitative ionic current. After establishing a gigohm seal, all recordings were made in the conventional whole-cell configuration. The external recording solution (150 mM NaCl, 10 mM KCl, 10 mM HEPES, 1 mM MgCl2, 3 mM CaCl2, 10 mM glucose, pH adjusted to 7.4 with NaOH) and the internal recording solution (20 mM KCl, 120 mM KF, 10 mM HEPES, 10 mM EGTA, 5 mM NaATP, pH adjusted to 7.2 with KOH) were used. During the experiment, 0.1% (v / v) BSA was included as a carrier in all external recording solutions. Using a voltage protocol, currents were elicited from a holding potential of -80 mV, which switched the voltage to 30 mV every 15 seconds for 500 milliseconds.

[0211] The concentration-response relationship was established by cumulatively applying seven increasing concentrations of the test sample to individual cells, with a recording period of 2 minutes for each compound application.

[0212] Potency was determined as the average charge of the last three scans starting from the cursor position at the end of each concentration application period. The percent inhibition for each test dose application period was calculated as the reduction in the average cursor value (charge) relative to the cursor value measured at the end of the carrier period and was used to calculate the IC from the concentration-response curve 50 . The results are shown in Table 3 below. Table 3 Example 3a: Inhibitory Activity of Kv1.3 Inhibitor Peptide on Human PBMC

[0213] Human peripheral blood mononuclear cells (PBMC) were used to evaluate the effect of the Kv1.3 inhibitor peptide on T cell activation, which was determined by IL-2 (cytokine) release after anti-CD3 stimulation.

[0214] Human PBMC were obtained from Precision for Medicine (Frederick, MD). Cells from 5 donors were used. Plate-bound anti-CD3 was used to stimulate a large number of T cells in the PBMC preparation. Briefly, a 96-well plate was coated with anti-CD3 antibody using 50 μL of a 0.5 μg / mL anti-CD3 solution diluted in 1× PBS at 37 °C for 2 hours. Thereafter, the plate was washed twice.

[0215] The peptides shown in Table 4a were diluted in medium (RPMI 1640, which had Glutamax-I, contained 10% v / v fetal bovine serum, 1% v / v penicillin-streptomycin solution) and added in a volume of 100 μL at concentrations ranging from 0.01 pM to 100 nM (ten-fold dilutions). Cyclosporin A (1 μg / ml) and Vm24 peptide (100 nM) were used as positive controls. Finally, 1×10 5 PBMCs were added to each well in a volume of 100 μL, bringing the final volume of each well to 200 μL. The plates were incubated in a 37 °C / 5% CO2 incubator for 20 to 24 hours. After centrifuging the plates, 25 μl of the supernatant was transferred to an IL-2 detection plate (MSD human IL-2 tissue culture kit, catalog number K151AHB-2), and IL-2 was measured as described by the manufacturer (Meso Scale Discovery, Rockville, Maryland, USA).

[0216] The results are shown in Table 4a as the geometric mean of the IC50 values obtained from anti-CD3-stimulated human PBMC assays. All values were derived from at least 4 replicates. Table 4a

[0217] Incubation in the presence of anti-CD3 antibody-activated hPBMCs and addition of the reference peptides resulted in a dose-dependent decrease in IL-2 secretion. The mean IC 50 values (calculated based on IL-2 release) were in the range of 0.05 nM to 0.4 nM. This was comparable to the IC 50 (IC 50 of 0.07 nM) observed with ShK186 and was approximately 1 / 100 to 1 / 10 of the IC 50 of Moka1 (which was less potent in inhibiting IL-2 secretion). There was no significant difference between the peptide and ShK186. Both ShK186 and the peptide were significantly lower than Moka1. In all experiments, cyclosporin completely blocked CD3-induced IL-2 release. Example 3b: Inhibitory activity of Kv1.3 inhibitor peptides on human PBMCs

[0218] Human peripheral blood mononuclear cells (PBMCs) were used to evaluate the effect of Kv1.3 inhibitor peptides on T cell activation, which was determined by IL-2 release after anti-CD3 stimulation.

[0219] Human PBMCs were obtained from Precision for Medicine (Frederick, MD). Cells from 5 donors were used. Plate-bound anti-CD3 was used to stimulate a large number of T cells in the PBMC preparation. Briefly, a 96-well plate was coated with anti-CD3 antibody with a 1 μg / ml anti-CD3 solution diluted in PBS at 5 °C for approximately 16 hours using 50 μl. The plate was then washed twice.

[0220] Subsequently, the peptides were diluted in medium (RPMI 1640 with Glutamax-I, containing 10% v / v fetal bovine serum, 1% v / v penicillin-streptomycin solution) and added in 50 μl volumes. The peptides shown in Table 4b were used at concentrations from 0.3 pM to 1000 nM (semi-logarithmic dilution, different starting concentrations). Cyclosporin A (1 μg / ml) and Vm24 peptide (100 nM) were used as positive controls.

[0221] Finally, 50,000 PBMCs in the same medium were added to each well in a 50 μl volume, bringing the final volume of each well to 100 μl. The plate was incubated in a 37 °C / 5% CO2 incubator for 20 to 24 hours. After centrifuging the plate, 25 μl of the supernatant was transferred to an IL-2 detection plate (MSD human IL-2 tissue culture kit, catalog number K151AHB-2), and IL-2 was measured as described by the manufacturer (Meso Scale Discovery, Rockville, Maryland, USA).

[0222] Results are shown in Table 4b as the geometric mean of the IC 50 values obtained from anti-CD3-stimulated human PBMC assays. All values were derived from at least 6 replicates. Table 4b

[0223] Incubation in the presence of anti-CD3 antibody-activated hPBMCs and addition of Kv1.3 inhibitor peptides resulted in a dose-dependent decrease in IL-2 secretion.

[0224] As shown in Table 4b, the mean IC 50 values (calculated based on IL-2 release) of the peptides were in the range of 0.01 nM to 0.09 nM. This is comparable to the IC 50 observed with ShK186 (IC 50 being 0.05 nM). This assay was performed using donors different from those used in Example 3a, so it was expected that the values of Shk-186 in the two sets of experiments would not be the same.

[0225] In all experiments, cyclosporine completely blocked anti-CD3-induced IL-2 release. Example 4: Inhibitory Activity of Kv1.3 Inhibitor Peptides in Rat Whole Blood

[0226] Rat whole blood was used to evaluate the potency of Kv1.3 inhibitor peptides on T cell activation, which was determined by IL-17A release after stimulation with thapsigargin. Addition of thapsigargin leads to activation of the signaling cascade, ultimately activating T cell proliferation and cytokine production, in which the Kv1.3 ion channel plays a key role. Thus, the activity of Kv1.3 inhibitors in primary cells can be measured in this experimental system.

[0227] Rat whole blood was obtained from healthy, naïve Lewis or Sprague-Dawley rats, which were terminally bled from the heart using heparinized blood collection tubes for collection. The peptides were diluted in assay buffer (DMEM + GlutaMAX) to 4× the final test concentration (GlutaMAX is a medium containing 3.97 mM L-alanine-L-glutamine (Gibco catalog number 61965026), supplemented with 25 mM HEPES buffer, 1 mM sodium pyruvate, 100 units / ml penicillin, 100 μg / ml streptomycin, and 0.05% casein from bovine milk (Sigma-Aldrich)) and 25 μl was added to the wells of a 96-well plate. Then 50 μl of rat whole blood was added and incubated at room temperature for at least 5 minutes to allow compound binding. Then 25 μl of 40 μM thapsigargin diluted in assay buffer was added to all wells of the assay plate to activate the cells, which were then incubated at 37 °C / 5% CO2 in a humidified chamber for 24 hours. The assay plate was centrifuged at 300 g for 10 minutes at 4 °C, and the supernatant was transferred to a new plate. The concentration of IL-17A released into the supernatant was measured using a rat IL-17A ELISA kit (Abcam catalog number ab214028) according to the manufacturer's recommendations. The samples were diluted 2.5-fold by transferring 20 μl of the supernatant to the wells on an ELISA plate from the detection kit containing 30 μl of buffer 75BS.

[0228] Data from peptides that caused inhibition of IL-17A were normalized relative to full thapsigargin activation (no inhibitor added) and non-activated controls (assay buffer added instead of thapsigargin) to calculate IC 50 .

[0229] The results are shown in Table 5, expressed as IC 50 , with standard deviation (IC 50_SD). All values are from at least 2 replicates. Ex vivo biological effects showed a correlation with peptide potency. Table 5 Example 5: Inhibitory Activity of Kv1.3 Inhibitor Peptides in Human Whole Blood

[0230] Human whole blood was used to evaluate the potency of Kv1.3 inhibitor peptides on T cell activation, which was determined by the release of cytokines IFN-γ, IL-2, and IL-17A after stimulation with thapsigargin. Adding thapsigargin activates the signaling cascade, leading to T cell proliferation and cytokine production, in which the Kv1.3 ion channel plays a key role. Therefore, the activity of Kv1.3 inhibitors in human primary T cells can be measured in this experimental system.

[0231] Human whole blood was obtained from healthy blood donors and collected using heparin sodium blood collection tubes (Becton, Dickinson and Company (BD), catalog number 367876) after informed consent. The test peptides were diluted to 4× the final test concentration in assay buffer (Dulbecco's Modified Eagle Medium (DMEM) with high glucose and GlutaMAX (3.97 mM L-alanine-L-glutamine) (Gibco catalog number 61965026), supplemented with 25 mM HEPES buffer, 1 mM sodium pyruvate, 100 units / ml penicillin, 100 μg / ml streptomycin, and 0.05% casein from bovine milk (Sigma-Aldrich catalog number C4765)), and 25 μl was added to the wells of a 96-well tissue culture plate. Then 50 μl of human whole blood was added and incubated at room temperature for at least 5 minutes to allow compound binding. Then 25 μl of 40 μM thapsigargin diluted in assay buffer was added to all wells of the assay plate to activate the cells, and then incubated in a humidified chamber at 37°C / 5% CO2 for 24 hours. The assay plate was centrifuged at 300 g for 10 minutes at 4°C, and the supernatant was transferred to a new plate. The concentrations of IFN-γ, IL-2, and IL-17A released into the supernatant were measured using a 3-plex human cytokine detection kit (MSD human U-Plex IFN-γ, IL-2, and IL-17A kit; Meso Scale Discovery catalog number K15067L-2), and all three cytokines were measured using an MSD MESOQuickPlex SQ 120 instrument according to the manufacturer's instructions (Meso Scale Discovery, Rockville, Maryland, USA).

[0232] To determine the inhibitory potency against the compound, cytokine concentration data was fitted using a three-parameter logistic dose response model based on the equation Y = Bottom + (Top - Bottom) / (1 + IC50 / X), where Y is the measured cytokine concentration, X is the compound concentration and Top, Bottom and IC 50 are the parameters fitted using the software Graphpad Prism version 5.04. The IC 50 value calculated from the concentration-response curve represents the compound concentration that produces a half-maximal inhibitory response between the basal (bottom) and maximal (top) responses.

[0233] The results are shown in Table 6, expressed as IC 50 . All values are from at least 2 replicate experiments. All tested Kv1.3 blockers were able to inhibit cytokine production from human T cells present in phorbol myristate acetate-stimulated human whole blood. Table 6 Example 6: Pharmacokinetic Characteristics of Kv1.3 Inhibitors

[0234] Male Sprague Dawley or Wistar rats (weighing approximately 250 g to 350 g) were given a single subcutaneous (s.c.) injection of each peptide to be tested.

[0235] After s.c. administration of the selected peptide (dose 70 nmol / kg, dosing volume 2 or 5 mL / kg), blood samples were taken at 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 min, 2 hours, 3 hours, 4 hours after dosing. At each sampling time point, samples were taken from the rats by sublingual bleeding or by tail clipping. After the last sampling, the rats were sacrificed by anesthesia with O2 / CO2. The dosing vehicle was 10 mM phosphate, 0.8% NaCl, 0.05% polysorbate 20 (pH 6.0).

[0236] Plasma samples were analyzed by liquid chromatography tandem mass spectrometry (LC-MS / MS) after solid phase extraction (SPE). In Phoenix WinNonlin 6.4 or higher, non-compartmental methods were used to calculate pharmacokinetic parameters using the mean plasma concentration. Plasma terminal elimination half-life (T 1 / 2) was determined to be ln(2) / λz, where λz is the magnitude of the slope of the log-linear regression of the log concentration versus time curve during the terminal phase. AUC inf is the area under the plasma concentration-time curve extrapolated to infinity (AUC inf = AUC last + C last / λz, where C last is the last observed plasma concentration). Cmax is the maximum concentration observed, which occurs at Tmax.

[0237] The results for some exemplary peptides are shown in Table 7. Table 7 Example 7: Effect of treatment with Kv1.3 inhibitor peptides on the rat keyhole limpet hemocyanin (KLH) ear inflammation model

[0238] A classical delayed-type hypersensitivity (DTH) reaction was induced in one ear of the rats. Briefly, male Lewis rats, 8 to 10 weeks old, were immunized subcutaneously (SC) at the base of the tail on day -7 with 200 μL of keyhole limpet hemocyanin (KLH) (from Sigma, catalog number H7017) (4 mg / mL) emulsified in complete Freund’s adjuvant (CFA) (Difco, catalog number 263810). On day 0, the rats were challenged intradermally in the left ear with 40 μL of KLH / 0.9% NaCl (2 mg / mL). After ear challenge, the rats developed a T cell-dependent inflammation in the KLH-challenged left ear. The right ear remained un-inflamed and served as a control.

[0239] The ability of Kv1.3 inhibitor peptide treatment to reduce the DTH ear swelling response was investigated by comparing the responses in rats treated with vehicle (n = 8 to 10 rats / group) with those treated with Kv1.3 inhibitor peptides. Twenty-four hours before KLH ear challenge, vehicle or peptide dissolved in vehicle (2 mL / kg) was administered SC. The test doses of the peptide were 50, 70, or 100 nmol / kg. The test vehicle was 10 mM phosphate, 0.8% w / v NaCl, 0.05% w / v polysorbate 20, pH 6. In all experiments, cyclosporine (CsA) was included as a positive study control. Cyclosporine (Sandimmune 100 mg / mL oral solution, Novartis) (10 mg / kg) was administered orally one hour before KLH ear challenge and again 6 hours after KLH ear challenge.

[0240] As the primary readout of efficacy, the area under the curve (AUC) of the Δ ear thickness (mm) was calculated for each animal from 0 to 48 hours after induction of the ear DTH response, where the change (D) was calculated as follows: left ear thickness - right ear thickness. These results were then used to calculate the percent inhibition of ear thickness by Kv1.3 inhibitor treatment: Percent inhibition: ((1 - (individual ΔAUC Kv1.3 inhibitor / mean ΔAUC vehicle group)) × 100. Results were calculated as percent inhibition + / - standard deviation (SD) and are shown in Tables 8 and 9. Table 8 Exp Dose (nmol / kg) Ptd3 Ptd42 Ptd53 CsA* #1 70 38.8(+ / -9.7) 46.9(+ / -10.9) - 71.4(+ / -4.8) #2 70 25.0(+ / -12.5) 27.7(+ / -11.5) - 76.8(+ / -13.4) #3 50 37.3(+ / -13.8) 22.2(+ / -11.1) - 65.1(+ / -4.5) #4 100 - 41.9(+ / -12.5) 25.1(+ / -6.3) 63.4(+ / -5.9) Table 9 Example 8: Prolonged effect of treatment with a Kv1.3 inhibitor peptide on the KLH-induced delayed type hypersensitivity (DTH) model Animal Care Committee

[0241] The animal care facility used was AAALAC-accredited. Female Lewis rats weighing 180 ± 20 g at arrival were used in this study. After arrival at the animal facility, all animals were evaluated for general health status. A 1-day acclimation period was allowed before the start of the study. Housing environment

[0242] The animals were housed under standardized environmental conditions. The rats were housed in top-opening cages with 6 animals per cage. Standard certified commercial rodent diet was provided ad libitum. Tap water was provided ad libitum at all times. The diet and water were considered to be free of known contaminants that would interfere with the objectives of this study. Each cage was labeled with the corresponding group, indicating the treatment and identity of the animals housed in the cage.

[0243] The animal room was maintained at a controlled temperature of 20 to 24 °C and a relative humidity of 30% to 70%. The controlled lighting system ensured 12 hours of light and 12 hours of darkness per day for the animals. Adequate ventilation was maintained at 15 air changes per hour. Research details

[0244] This study was conducted to investigate the duration of action of a Kv1.3 inhibitor (peptide 100) on ear inflammation in a rat KLH-induced DTH model.

[0245] Healthy male Lewis rats, 8 to 9 weeks old, were immunized subcutaneously (SC) at the base of the tail on day 0 with 200 μL of keyhole limpet hemocyanin (KLH) (from Sigma, catalog number H7017) (4 mg / mL) emulsified in complete Freund's adjuvant (CFA) (Difco, catalog number 263810).

[0246] To induce local inflammation mediated by KLH-specific T cells, rats were intradermally challenged in the left ear with 40 μL of KLH / 0.9% NaCl (2 mg / mL) at the specified time points (days 7, 9, 11, or 13 after immunization). Ear edema was measured 24 to 48 hours later, and the right ear was left untreated and used as a control. A schematic overview of the protocol is depicted in Figure 1 .

[0247] The ability of Kv1.3 inhibitor treatment to reduce the DTH ear swelling response was investigated by comparing the responses in vehicle-treated rats (n = 8 rats / group) with those in rats treated with peptide 100. On day 6, the vehicle or peptide 100 (2 mL / kg) dissolved in the vehicle was administered SC. The test dose of the Kv1.3 inhibitor was 300 nmol / kg. The test vehicle was 10 mM phosphate, 0.8% w / v NaCl, 0.05% w / v polysorbate 20, pH 6. Cyclosporine A (CsA) was included as a positive study control. Cyclosporine (Sandimmune 100 mg / mL oral solution, Novartis) (10 mg / kg) was administered orally one hour before KLH ear challenge on day 7 and again 6 hours after KLH ear challenge.

[0248] As a readout of efficacy, the thickness of the induced ear was measured 24 and 48 hours after the respective ear challenges and compared to the vehicle control. The ear thickness 24 hours after challenge is shown in Table 10 and Figure 2 , and the ear thickness 48 hours after challenge is shown in Table 11 and Figure 3 . The results are also summarized below.

[0249] For animals challenged on day 7 (i.e., 1 day after treatment), when measuring ear swelling 24 hours after challenge (i.e., 2 days after treatment) (peptide 100: 0.676 ± 0.017 mm; vehicle: 0.796 ± 0.03 mm; CsA: 0.557 ± 0.021 mm; p < 0.0001) and when measuring ear swelling 48 hours after challenge (i.e., 3 days after treatment) (peptide 100: 0.623 ± 0.015 mm; vehicle: 0.733 ± 0.02 mm; CsA: 0.539 ± 0.018 mm; p < 0.0001), peptide 100 and CsA significantly reduced ear swelling compared to the vehicle control.

[0250] When ear challenges were performed on rats on day 9 (i.e., 3 days after treatment), ear swelling was measured 24 hours after the challenge (i.e., 4 days after treatment) (peptide 100: 0.689 ± 0.031 mm; vehicle: 0.761 ± 0.04 mm; p = 0.0011) and 48 hours after the challenge (i.e., 5 days after treatment) (peptide 100: 0.659 ± 0.034 mm; vehicle: 0.729 ± 0.02 mm; p = 0.0002), and ear swelling was significantly reduced compared to the vehicle control.

[0251] When ear challenges were performed on rats on day 11 (i.e., 5 days after treatment), ear swelling was measured 24 hours after the challenge (i.e., 6 days after treatment) (peptide 100: 0.681 ± 0.023 mm; vehicle: 0.784 ± 0.025 mm; p < 0.0001) and 48 hours after the challenge (i.e., 7 days after treatment) (peptide 100: 0.653 ± 0.013 mm; vehicle: 0.746 ± 0.41 mm; p < 0.0001), and ear swelling was significantly reduced compared to the vehicle control.

[0252] For ear challenges on day 13 (i.e., 7 days after treatment), ear swelling was measured 24 hours after the challenge (i.e., 8 days after treatment) (peptide 100: 0.729 ± 0.038 mm; vehicle: 0.767 ± 0.039 mm; p = 0.0696) and 48 hours after the challenge (i.e., 9 days after treatment) (peptide 100: 0.699 ± 0.028 mm; vehicle: 0.720 ± 0.036 mm; p = 0.2256), and the reduction was not significant.

[0253] In summary, peptide 100 administered on day 6 reduced KLH-induced ear swelling when measured 24 hours after challenges on days 7, 9, and 11 (i.e., ear swelling reduction was observed on days 2, 4, and 6 after treatment) and when measured 48 hours after challenges (i.e., ear swelling reduction was measured on days 3, 5, and 7 after treatment). Table 10: Mean ear thickness (mm) ± SD of KLH-induced ears measured 24 hours after challenge Table 11: Mean ear thickness (mm) ± SD of KLH-induced ears measured 48 hours after challenge Example 9: Different dosing regimens of Kv1.3 inhibitors in a rat collagen-induced arthritis (CIA) model Peptide and vehicle formulations

[0254] Vehicle: 10 mM phosphate pH 6 + 0.8% NaCl + 0.05% polysorbate 20

[0255] Peptide: Peptide 100 was formulated in the vehicle at 50 nmol / mL. The dosing volume was adjusted individually according to the body weight of each animal to achieve a target dose of 100 nmol / kg of Peptide 100. Animals

[0256] The animal care facilities used were AAALAC - accredited. Female Lewis rats weighing 180 ± 20 g upon arrival were used in this study. After arrival at the animal facility, all animals were evaluated for general health status. A 1 - day acclimation period was allowed before the start of the study.

[0257] The animals were housed under standardized environmental conditions. The rats were housed in top - opened cages with 6 animals per cage. Standard certified commercial rodent diet was provided ad libitum. Tap water was always provided ad libitum. It was considered that there were no known contaminants in the diet and water that would interfere with the objectives of this study. Each cage was labeled with the corresponding group, indicating the treatment and identity of the animals housed in the cage.

[0258] The animal room was maintained at a controlled temperature of 20 to 24 °C and a relative humidity of 30% to 70%. The controlled lighting system ensured 12 - hour light and 12 - hour dark for the animals every day. Sufficient ventilation was maintained at 15 air changes per hour. Immunization of Rats

[0259] Based on their body weight, they were divided into groups of 9 animals each. All animals were challenged on Day 1 with porcine type II collagen together with incomplete Freund's adjuvant (0.2 mg / 0.2 mL / rat, subcutaneously at the base of the tail) and boosted on Day 7 (0.1 mg / 0.1 mL / rat, s.c.). In this study, dexamethasone was administered at 0.3 mg / kg daily PO as a positive control. Administration of Test Peptide and Vehicle

[0260] From Day 12 to Day 30, Peptide 100 was administered subcutaneously to the flanks daily (QD), every three days (Q3D), or every five days (Q5D) as a single bolus. On the days when the animals did not receive the peptide, only the vehicle was administered to the animals. The peptide and the vehicle were administered at a volume of 2 mL / kg. The dosing volume was adjusted individually according to the body weight of each animal to achieve the target dose of Formulation 1 at 100 nmol / kg. Disease Scoring

[0261] The disease was evaluated at days 1, 7, 10, 12 (before dosing), 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24, 26, and 28 before peptide treatment and at one hour after treatment on day 30 using a severity qualitative scoring system (see below, maximum score of 16).

[0262] The disease severity score was based on the following:

[0263] The ability of Kv1.3 inhibitor treatment to reduce the chronic inflammation of arthritis was investigated by comparing the responses in rats treated with vehicle (n = 9 animals / group) with those treated with peptide 100 daily, every three days, or every five days. The severity of arthritis was determined by summing the disease scores of all four paws of each animal from day 12 to day 30. These results are shown in Table 12. Treatment with peptide 100 at all dosing regimens led to a reduction in the mean clinical score for all dosing regimens. Table 12

[0264] The mean clinical score of the front paws was analyzed separately and shown in Figure 4 . All three dosing regimens (daily, every three days, and every five days) were able to delay the disease in the front paws. Example 10: Determination of the maximum effective dose of treatment with a Kv1.3 inhibitor peptide in a KLH-induced delayed-type hypersensitivity (DTH) model

[0265] This study was conducted as described in Example 8, except that rats were intradermally challenged in the left ear with 40 μL of KLH / 0.9% NaCl (2 mg / mL) only on days 7 or 11 after immunization (i.e., days 1 and 5 after treatment). A schematic overview of this protocol is depicted in Figure 5 .

[0266] The dose at which Kv1.3 inhibitor treatment can reduce the DTH ear swelling response was investigated by comparing the responses in rats treated with vehicle (n = 8 / group) with those treated with different doses of peptide 100. On day 6 after immunization, vehicle or peptide 100 (2 mL / kg) dissolved in vehicle was administered SC. The test doses of the Kv1.3 inhibitor were 10, 100, 300, or 700 nmol / kg. The test vehicle was 10 mM phosphate, 0.8% w / v NaCl, 0.05% w / v polysorbate 20, pH 6.

[0267] As a readout of potency, the thickness of the induced ear was measured 24 and 48 hours after each ear challenge and compared to vehicle control. The ear thickness 24 hours after challenge is shown in Table 13 and Figure 6 and the ear thickness 48 hours after challenge is shown in Table 14 and Figure 7 and is also summarized below.

[0268] For animals challenged on Day 7 (i.e., 1 day after treatment), when measuring ear swelling 24 hours after challenge (i.e., 2 days after treatment) (peptide 100: 0.704 ± 0.018 mm, p = 0.1216; vehicle: 0.721 ± 0.021 mm; CsA: 0.528 ± 0.032 mm, p < 0.0001) and when measuring ear swelling 48 hours after challenge (i.e., 3 days after treatment) (peptide 100: 0.652 ± 0.026 mm, p = 0.0008; vehicle: 0.694 ± 0.011 mm; CsA: 0.497 ± 0.021 mm, p < 0.0001), CsA showed reduced ear swelling compared to vehicle control.

[0269] In the case of peptide 100 at doses of 10, 100, 300 or 700 nmol / kg, when rats were ear challenged on Day 11 (i.e., 5 days after treatment), when measuring ear swelling 24 hours after challenge (i.e., 6 days after treatment) (peptide 100 at 10 nmol / kg: 0.807 ± 0.025 mm, p = 0.0495; 100 nmol / kg: 0.777 ± 0.019 mm, p = 0.0015; 300 nmol / kg: 0.772 ± 0.009 mm, p = 0.0005; 700 nmol / kg: 0.771 ± 0.018 mm, p = 0.0007; vehicle: 0.848 ± 0.047 mm) and when measuring ear swelling 48 hours after challenge (i.e., 7 days after treatment) (peptide 100 at 10 nmol / kg: 0.761 ± 0.025 mm, p = 0.0009; 100 nmol / kg: 0.713 ± 0.034 mm, p < 0.0001; 300 nmol / kg: 0.684 ± 0.016 mm, p < 0.0001; 700 nmol / kg: 0.671 ± 0.011 mm, p < 0.0001; vehicle: 0.819 ± 0.03 mm), ear swelling was significantly reduced compared to vehicle control.

[0270] In summary, the maximum effect was obtained with peptide 100 at a dose of 300 nmol / kg in the KLH-induced DTH model. Table 13: Mean ear thickness (mm) ± SD of KLH-induced ears measured 24 hours after challenge Table 14: Mean ear thickness (mm) ± SD of KLH-induced ears measured 48 hours after attack Example 11: Determination of the minimum effective dose of treatment with a Kv1.3 inhibitor peptide in a KLH-induced delayed-type hypersensitivity (DTH) model

[0271] This study was conducted as described in Example 8, except that rats were intradermally challenged in the left ear with 40 μL of KLH / 0.9% NaCl (2 mg / mL) only on days 7 or 11 after immunization (i.e., days 1 and 5 after treatment). A schematic overview of this protocol is depicted in Figure 5 .

[0272] The dose at which Kv1.3 inhibitor treatment can reduce the DTH ear swelling response was investigated by comparing the responses in rats treated with vehicle (n = 8 rats / group) with those treated with different doses of peptide 100. On day 6, vehicle or peptide 100 (2 mL / kg) dissolved in vehicle was administered SC. The test doses of the Kv1.3 inhibitor were 1, 3, 10, 30, or 100 nmol / kg. The test vehicle was 10 mM phosphate, 0.8% w / v NaCl, 0.05% w / v polysorbate 20, pH 6.

[0273] As a readout of potency, the thickness of the induced ears was measured 24 and 48 hours after the respective ear challenges and compared to the vehicle control. The ear thickness 24 hours after challenge is shown in Table 15 and Figure 8 and the ear thickness 48 hours after challenge is shown in Table 16 and Figure 9 . The results are also summarized below.

[0274] For animals challenged on day 7 (i.e., 1 day after treatment), when ear swelling was measured 24 hours after challenge (i.e., 2 days after treatment) (peptide 100: 0.736 ± 0.026 mm, p < 0.0001; vehicle: 0.849 ± 0.032 mm; CsA: 0.601 ± 0.033 mm, p < 0.0001) and when ear swelling was measured 48 hours after challenge (i.e., 3 days after treatment) (peptide 100: 0.671 ± 0.022 mm, p < 0.0001; vehicle: 0.777 ± 0.036 mm; CsA: 0.551 ± 0.021 mm, p < 0.0001), the ear swelling was significantly reduced with CsA compared to the vehicle control.

[0275] In the case of peptide 100 at a dose of 1, 3, 10, 30 or 100 nmol / kg, when rats were subjected to ear challenge on day 11 (i.e., 5 days after treatment), ear swelling was measured 24 hours after the challenge (i.e., 6 days after treatment) (peptide 100 at 1 nmol / kg: 0.872 ± 0.037 mm, p = 0.0872; 3 nmol / kg: 0.814 ± 0.014 mm, p < 0.0001; 10 nmol / kg: 0.796 ± 0.043 mm, p < 0.0001; 30 nmol / kg: 0.754 ± 0.017 mm, p < 0.0001; 100 nmol / kg: 0.751 ± 0.009 mm, p < 0.0001; vehicle: 0.902 ± 0.028 mm) and when ear swelling was measured 48 hours after the challenge (i.e., 7 days after treatment) (peptide 100 at 1 nmol / kg: 0.854 ± 0.038 mm, p = 0.1953; 3 nmol / kg: 0.814 ± 0.019 mm, p = 0.0001; 10 nmol / kg: 0.794 ± 0.036 mm, p = 0.0001; 30 nmol / kg: 0.736 ± 0.03 mm, p < 0.0001; 100 nmol / kg: 0.731 ± 0.016 mm, p < 0.0001; vehicle: 0.876 ± 0.027 mm), ear swelling was significantly reduced compared to the vehicle control.

[0276] In summary, a dose of 3 nmol / kg of peptide 100 is the minimum dose to achieve an effect in the KLH-induced DTH model. Table 15: Mean ear thickness (mm) ± SD of KLH-induced ears measured 24 hours after challenge Table 16: Mean ear thickness (mm) ± SD of KLH-induced ears measured 48 hours after challenge

[0277] All publications mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the methods and systems described in the present invention will be apparent to those skilled in the art and do not depart from the scope and spirit of the present invention. Although the present invention has been described in connection with some specific preferred embodiments, it should be understood that the claimed invention should not be unduly limited to such specific embodiments. In fact, various modifications of the described ways of implementing the present invention that are apparent to those skilled in the art of biochemistry, molecular biology or related fields are intended to be within the scope of the appended aspects.

Claims

1. A Kv1.3 inhibitor, or a pharmaceutically acceptable salt thereof, for use in a method of treating or preventing a disease or disorder in a subject, wherein the Kv1.3 inhibitor comprises or consists of a peptide that comprises the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof or consists of the sequence QMDMRCSASVECKQKCLKAIGSIFGKCMNKKCKCYPR (SEQ ID NO 1) or a variant thereof, wherein the variant (a) has at least 70% sequence identity with SEQ ID NO 1, and / or (b) differs from SEQ ID NO 1 by a total of at most nine substitutions, insertions and / or deletions, and wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

2. The Kv1.3 inhibitor or pharmaceutically acceptable salt of the application according to claim 1, wherein the IC 50 of the Kv1.3 inhibitor against the human Kv1.3 potassium channel is 400 nM or less, preferably 300 nM or less, preferably 50 nM or less, preferably 15 nM or less, preferably 10 nM or less, preferably 5 nM or less, preferably 2 nM or less.

3. The Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to claim 1 or claim 2, wherein the variant has at least 70% sequence identity with SEQ ID NO 1, preferably at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with SEQ ID NO 1.

4. The Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 3, wherein the peptide comprises the following amino acids: The amino acid at position 6 is C; The amino acid at position 12 is C; The amino acid at position 16 is C; The amino acid at position 27 is C; The amino acid at position 32 is C; and The amino acid at position 34 is C; Preferably, further wherein: The amino acid at position 24 is F; The amino acid at position 25 is G; The amino acid at position 29 is N; and The amino acid at position 31 is K.

5. The Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 4, wherein the variant differs from SEQ ID NO 1 by a total of at most nine substitutions, insertions and / or deletions, preferably a total of at most eight, at most seven, at most six, at most five, at most four, at most three or at most two substitutions, insertions and / or deletions or a total of one substitution, insertion or deletion.

6. The Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 5, wherein any substitution or deletion in the variant of SEQ ID NO 1 is at an amino acid position selected from positions 1 to 5, 7 to 11, 13 to 15, 17 to 23, 26, 28, 30, 33 and 35 to 37 of SEQ ID NO 1, preferably at an amino acid position selected from positions 1, 2, 3, 4, 5, 7, 14, 18, 19, 28 and 37 of SEQ ID NO 1.

7. The Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 6, wherein the peptide comprises the following amino acids: The amino acid at position 1 is H, N, P, p, Q, S, V or Y, or is absent; The amino acid at position 2 is I, M or Nle, or is absent; The amino acid at position 3 is D, E or S, or is absent; The amino acid at position 4 is E, L, M, Nle, S or V, or is absent; The amino acid at position 5 is R or K, or is absent; The amino acid at position 7 is E, F, H, K, Orn, R, S, Y, 2,3-diaminopropionyl, 2,4-diaminobutyryl or 2-amino-3-guanidinopropionyl; The amino acid at position 8 is A, H, I, L, S or Y; The amino acid at position 9 is F, L, P, S, Orn, V, Abu or 2,3-diaminopropionyl; The amino acid at position 10 is K, P, Q, R or V; The amino acid at position 11 is E or Q; The amino acid at position 13 is A, E, G, K, L, Q or V; The amino acid at position 14 is E, K, L, Q, V or 2-amino-5-carboxypentanoyl; The amino acid at position 15 is K, L, P, S; The amino acid at position 17 is K, L, R or Y, or is absent; The amino acid at position 18 is A, D, G, K, Q, hQ, V or Y, or is absent; The amino acid at position 19 is A, K, R or Y, or is absent; The amino acid at position 20 is E, I, R or Y, or is absent; The amino acid at position 21 is E, G, H or R; The amino acid at position 22 is C, R or S; The amino acid at position 23 is G, I, K, P or R; The amino acid at position 26 is K or hK; The amino acid at position 28 is M or Nle; The amino acid at position 30 is G or K; The amino acid at position 33 is H, K, R or V; The amino acid at position 35 is Y, F(4-F), F(4-CH3), F(4-NO2) or F(4-NH2); The amino acid at position 36 is Q or P, or is absent; and / or The amino acid at position 37 is C, G, R, S or (4-amino-5-hydroxypentyl)guanidine, or is absent.

8. A Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 7, wherein the peptide comprises one or consists of one of the following sequences:

9. A Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 8, wherein the peptide is selected from the following peptides:

10. A Kv1.3 inhibitor or pharmaceutically acceptable salt for use according to any one of claims 1 to 9, wherein the disease or disorder (a) is an inflammatory disease or disorder, preferably, wherein the inflammatory disease or disorder is selected from autoimmune diseases, allergic or hypersensitivity reactions, allograft rejection, transplant rejection, graft-versus-host disease, hay fever, asthma, allergic reactions, allergic rhinitis, urticaria, eczema, alopecia areata, dermatomyositis, inclusion body myositis, polymyositis, ankylosing spondylitis, vasculitis, arthritis (including rheumatoid arthritis, osteoarthritis, psoriatic arthritis), Sjogren's syndrome, systemic lupus erythematosus (SLE), uveitis, inflammatory fibrosis (such as scleroderma, pulmonary fibrosis, liver cirrhosis), chronic obstructive pulmonary disease (COPD), hepatitis, chronic inflammatory demyelinating polyneuropathy, inflammatory bowel disease, colitis (such as Crohn's disease and ulcerative colitis), erythema, thyroiditis, psoriasis, atopic dermatitis, allergic contact dermatitis, scleroderma, glomerulonephritis, inflammatory bone resorption, multiple sclerosis and type 1 diabetes; or (b) is a metabolic disease or disorder; or (c) is selected from obesity, obesity-related inflammation, obesity-related gallbladder diseases and obesity-induced sleep apnea; or (d) is caused by or associated with impaired glucose control, preferably, wherein the disease or disorder is selected from metabolic syndrome, insulin resistance, glucose intolerance, prediabetes, elevated fasting blood glucose and type 2 diabetes; or (e) is a smooth muscle proliferative disorder, preferably, wherein the smooth muscle proliferative disorder is restenosis; or (f) is a neuroinflammatory or neurodegenerative disease or disorder, preferably, wherein the neuroinflammatory or neurodegenerative disease or disorder is selected from Alzheimer's disease, multiple sclerosis (MS), Parkinson's disease and amyotrophic lateral sclerosis (ALS); or (g) is a cancer, preferably, wherein the cancer is breast cancer, prostate cancer or lymphoma, preferably, wherein the lymphoma is non-Hodgkin lymphoma (NHL), preferably, wherein the NHL is selected from diffuse large B-cell lymphoma, follicular lymphoma, Burkitt lymphoma, immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma, mantle cell lymphoma, mycosis fungoides, anaplastic large cell lymphoma, peripheral T-cell lymphoma, precursor T lymphoblastic lymphoma and Sezary syndrome.

11. The Kv1.3 inhibitor, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 9, for use in a method for: (a) inhibiting or reducing inflammation; or (b) inhibiting weight gain, promoting weight loss, reducing overweight or treating obesity; wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 8 days.

12. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to any one of claims 1 to 11, wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 to 7 days, once every 2 to 6 days, once every 2 to 5 days, once every 2 to 4 days, once every 2 to 3 days, once every 3 to 8 days, once every 3 to 7 days, once every 3 to 6 days, once every 3 to 5 days, once every 3 to 4 days, once every 4 to 8 days, once every 4 to 7 days, once every 4 to 6 days, once every 4 to 5 days, once every 5 to 8 days, once every 5 to 7 days, once every 5 to 6 days, once every 6 to 8 days, once every 6 to 7 days, or once every 7 to 8 days.

13. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to any one of claims 1 to 12, wherein the method comprises administering the Kv1.3 inhibitor to the subject once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, or once every 8 days, preferably administering the Kv1.3 inhibitor to the subject once every 7 days.

14. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to any one of claims 1 to 13, wherein the method comprises administering the Kv1.3 inhibitor to the subject by injection, preferably by subcutaneous injection.

15. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to any one of claims 1 to 14, wherein the method comprises administering the Kv1.3 inhibitor to the subject at a dose of 0.1 mg to 30.0 mg.

16. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to any one of claims 1 to 15, wherein the method comprises administering the Kv1.3 inhibitor to the subject at a dose of 10 nmol / kg to 400 nmol / kg or at a dose of 3 nmol / kg to 300 nmol / kg.

17. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to any one of claims 1 to 16, wherein the Kv1.3 inhibitor or the pharmaceutically acceptable salt is in the form of a composition, preferably wherein the composition is a pharmaceutical composition, and preferably, wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier, excipient, or vehicle.

18. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to claim 17, wherein the composition comprises one or more peptides, each of which peptides comprises or consists of a sequence independently selected from the following sequences:

19. A Kv1.3 inhibitor or a pharmaceutically acceptable salt thereof for use according to claim 17 or claim 18, wherein the composition comprises one or more peptides, which peptides are independently selected from the following peptide sequences:

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