WO2022232315A1 - Compounds - Google Patents

Abstract

The present invention provides synthetic peptides and substituted peptides that exhibit potent NaV1.7 inhibition. The present invention provides compounds, methods and compositions effective to image nerve anatomy or physiology, either by PET imaging or by fluorescent optical imaging, to treat pain by way of NaV1.7 inhibition, to treat itching by way of NaV1.7 inhibition, and to treat cancer by way of NaV1.7 inhibition.

Description

Compounds

Claim of Priority

This application claims priority under 35 U.S.C. §119(e) to U.S. Patent Application Serial No. 63/180,208, filed on April 27, 2021, which is incorporated herein by reference in its entirety.

Technical Field

The present invention relates to radio-labeled compounds useful in the imaging of pain, fluorophore-containing compounds useful in the imaging of peripheral neurons, compounds useful in the treatment and management of pain, and compounds useful in the treatment of cancer.

Background Art

In 2006, it was reported that nonsense mutations in SCN9A lead to a congenital inability to sense pain. This gene encodes the a-subunit of the voltage-gated sodium channel NaV1.7. Loss of function of NaV1.7 does not seem to otherwise impair signal transduction. Intelligence and other sensory functions, such as sensing heat, touch, pressure and motor function, were not impacted. NaV1.7 is expressed in peripheral sensory neurons, particularly nociceptive small-diameter dorsal root ganglia. NaV1.7 amplifies membrane potential, where neuronal firing initiates downstream cascades that ultimately result in pain sensation. Gain/loss of function mutants show thatNaV1.7 is a critical regulator of pain. Increased expression occurs concurrently with peripheral neuropathy, therefore NaV1.7 is a direct functional target for imaging pain, peripheral neurons, and treating pain. A screen of venoms from 52 species of spider and 12 species of scorpion produced a peptide, Hspla, that was capable of potently and selectively inhibiting NaV1.7. This peptide represents the parent compound upon which all modified peptides disclosed herein are based.

NaV1.7 is also known to have activity beyond pain regulation, including in urticaria. In 2014, it was reported that some NaV1.7 inhibitors have the ability to treat urticaria. Therefore, NaV 1.7 is a direct target for the treatment of urticaria.

NaV1.7 has also been implicated in the pathology of several cancers. In 2010, it was reported that NaV1.7 over expression is associated with metastatic ovarian cancer cells. In 2016, it was reported thatNaV1.7, which is over expressed in gastric cancer, promotes progression of gastric cancer, and that inhibition of NaV1.7 has the potential to inhibit progression of gastric cancer. Similar behavior has been observed in several other cancers. Therefore, NaV1.7 is a direct target for the treatment of certain cancers.

Summary of Invention

The present invention relates to a series of peptide based compounds for the imaging of pain, for the imaging of peripheral nerves, for the treatment of pain, for the treatment of itching, and for the treatment of cancer. These peptides are modified to enhance properties of the peptide, such as potency and/or specificity, or to provide for beneficial biophysical properties like pharmacokinetics and pharmacodynamics, or to add functionality like PET or optical imaging capabilities.

The invention includes modified peptides based on Hspla, a peptide isolated from the Peruvian Blue Tarantula. The modified peptides provide improved properties over the wild-type peptide including better overall stability and better stability when administered orally. The invention includes the administration of these peptides for pain management, urticaria management, and cancer treatment. The invention further includes the administration of Hspla peptide derivatives that have been modified for use in radio therapy, radio imaging (including PET), and fluorescence imaging.

The present technology provides NaV1.7 inhibiting peptides. These peptides are useful for treating pain, for locating and quantifying pain, for identifying the location of nerves in patients, for treating itching, and for treating cancer. In one embodiment, NaV1.7 inhibiting peptides exhibit enhanced pharmacological properties that allow them to be orally bioavailable for use as therapeutic agents to treat pain. In another embodiment, radiolabeled NaV1.7 inhibiting peptides are used to map where in a patient there is an over expression of NaV1.7, which can be correlated with the amount of pain being experienced by the patient, giving a clinician the ability to locate and quantify a patient's pain. In another embodiment, fluorescent labelled NaV1.7 inhibiting peptides can be used to map the location of nerves in a patient, which enables a surgeon to avoid nerves during surgery. In another embodiment, NaV1.7 inhibiting peptides or radiolabelled NaV1.7 inhibiting peptides are used as a therapeutic agent to treat cancer. In another embodiment, NaV1.7 inhibiting peptides exhibit enhanced pharmacological properties that allow them to be orally bioavailable for use as a therapeutic agent to treat itching. As used herein, "about" will be understood by persons of ordinary skill in the art and will vary depending on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art, given the context in which it is used, "about" will mean up to plus or minus 10% of the particular term. As will be understood by one skilled in the art, for any and all purposes, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof.

Pharmaceutically acceptable salts of compounds described herein are within the scope of the present technology and include acid or base addition salts which retain the desired pharmacological activity and are not biologically undesirable.

Those of skill in the art will appreciate that compounds of the present technology may exhibit the phenomena of tautomerism, conformational isomerism, geometric isomerism and/or stereoisomerism. It should be understood that the present technology encompasses any tautomeric, conformational isomeric, stereochemical and/or geometric isomeric forms of the compounds having one or more of the utilities described herein, as well as mixtures of these various different forms.

"Cross-linking agents" refers to agents that include, but are not limited to, EGS (i.e., ethylene glycol bisfsuccinimidylsuccinate]), DSS (i.e., disuccinimidyl suberate), DMA (i.e., dimethyl adipimidate.2HCI), DTSSP (i.e., 3,3' dithiobis[sulfosuccinimidylpropionate])),

DPDPB (i.e., l,4-di-[3'-(2'-pyridyldithio)-propionamido]butane), BMH (i.e., bis- maleimidohexane), maleimidyl linkers, alkyl halide linkers, succinimidyl linkers, and the platinum cross-linking agents described in U.S. Pat. Nos.5, 580, 990; 5,985,566; and 6,133,038 of Kreatech Biotechnology, B.V., Amsterdam, The Netherlands

The peptides of the present invention may also be stapled or stitched peptides. A "stapled" peptide refers to a peptide, typically containing an alpha-helix, that contains a covalent linkage between two side chains of amino acids that make up the peptide. A stapled peptide may form a peptide macrocycle. A peptide that has multiple peptide staple points may be referred to as a "stitched" peptide. In a preferred embodiment, stapling via ring-closing metathesis is used. In another preferred embodiment, stapling occurs via a boron containing amino acid side chain. In another preferred embodiment, a peptide staple is substituted for a disulfide bond in a peptide. Stereoisomers of compounds (also known as optical isomers) include all chiral, diastereomeric, and racemic forms of a structure, unless the specific stereochemistry is expressly indicated. Thus, compounds used in the present technology include enriched or resolved optical isomers at any or all asymmetric atoms as are apparent from the depictions. Both racemic and diastereomeric mixtures, as well as the individual optical isomers can be isolated or synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and these stereoisomers are all within the scope of the present technology.

The compounds of the present technology may exist as solvates, especially hydrates. Hydrates may form during manufacture of the compounds or compositions comprising the compounds, or hydrates may form over time due to the hygroscopic nature of the compounds. Compounds of the present technology may exist as organic solvates as well, including DMF, ether, and alcohol solvates among others. The identification and preparation of any particular solvate is within the skill of the ordinary artisan of synthetic organic or medicinal chemistry.

As used herein, the terms "subject," "individual," or "patient" can be an individual organism, a vertebrate, a mammal, or a human. "Mammal" includes a human, non-human primate, murine (e.g., mouse, rat, guinea pig, hamster), ovine, bovine, ruminant, lagomorph, porcine, caprine, equine, canine, feline, avians, etc. In a preferred embodiment, the mammal is feline or canine. In another preferred embodiment, the mammal is human.

The term "administering" a compound or composition to a subject means delivering the compound to the subject. "Administering" includes prophylactic administration of the compound or composition (i.e., before the disease and/or one or more symptoms of the disease are detectable) and/or therapeutic administration of the composition (i.e., after the disease and/or one or more symptoms of the disease are detectable). The methods of the present technology include administering one or more compounds or agents. If more than one compound is to be administered, the compounds may be administered together at substantially the same time, and/or administered at different times in any order. Also, the compounds of the present technology may be administered before, concomitantly with, and/or after administration of another type of drug or therapeutic procedure (e.g., surgery). As used herein, the term "amino acid" is used to refer to any organic molecule that contains at least one amino group and at least one carboxyl group where the at least one amino group is at the a position relative to the carboxyl group, where the amino acid is in the L- configuration. Naturally occurring amino acids include, for example, the twenty most common levorotatory (L,) amino acids normally found in mammalian proteins, i.e., alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (lie), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan, (Trp), tyrosine (Tyr), and valine (Val). Naturally-occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, g-carboxyglutamate, and O-phosphoserine. Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the lUPAC-IUB Biochemical Nomenclature Commission.

As used herein, the terms "polypeptide," "polyamino acid," "peptide," and "protein" are used interchangeably herein to mean a polymer comprising two or more amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres. Polypeptide refers to both short chains, commonly referred to as peptides, glycopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. Polypeptides include amino acid sequences modified either by natural processes, such as post-translational processing, or by chemical modification techniques that are well known in the art.

As used herein, "isolated" or "purified" polypeptide, peptide, or protein refers to polypeptide, peptide, or protein that is substantially free of cellular material or other contaminating polypeptides from the cell or tissue source from which the agent is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. For example, an isolated protein would be free of materials that would interfere with therapeutic uses of the agent.

"Treating," "treat," "treated," or "treatment" as used herein covers the treatment of a disease or disorder described herein in a subject, such as a human, and includes: (i) inhibiting a disease or disorder, i.e., arresting its development; (ii) relieving a disease or disorder, i.e., causing regression of the disorder; (iii) slowing progression of the disorder; and/or (iv) inhibiting, relieving, ameliorating, or slowing progression of one or more symptoms of the disease or disorder. Symptoms may be assessed by methods known in the art or described herein, for example, biopsy, histology, and blood tests to determine relevant enzyme levels, metabolites or circulating antigen or antibody (or other biomarkers), quality of life questionnaires, patient-reported symptom scores, and imaging tests.

"Ameliorate," "ameliorating," and the like, as used herein, refer to inhibiting, relieving, eliminating, or slowing progression of one or more symptoms.

As used herein, "prevention," "prevents," or "preventing" of a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder, symptom, or condition in the treated sample relative to a control subject, or delays the onset of one or more symptoms of the disorder or condition relative to the control subject.

Injuries to the peripheral nervous system represent a significant concern in surgical practice, and can occur during virtually any type of intervention. While the majority of peripheral nerve injuries occur in the upper limbs and are of traumatic origin, around 25% of patients suffering from neuropathic pain identified surgical intervention as the originating cause. Oncologic surgery in particular carries a considerable risk of peripheral nerve damage because of the distorted physiology around a malignant lesion and the need to achieve complete resection for oncologic control - which increases the likelihood of inadvertent injury.

A "conservative amino acid substitution variant" will be well understood by one of ordinary skill in the art. One of ordinary skill in the art understands amino acids may be grouped according to their physicochemical characteristics as follows:

(a) Non-polar amino acids: Ala(A) Ser(S) Thr(T) Pro(P) Gly(G) Cys (C);

(b) Acidic amino acids: Asn(N), Asp(D), Glu(E), Gln(Q);

(c) Basic amino acids: His(H), Arg(R), Lys(K);

(d) Hydrophobic amino acids: Met(M), Leu(L), Ile(I) ,Val(V); and

(e) Aromatic amino acids: Phe(F) Tyr(Y) Trp(W).

Substitutions of an amino acid in a peptide by another amino acid in the same group are referred to as a conservative substitution (and the resulting peptide a “conservative amino acid substitution variant”) and may preserve the physicochemical characteristics of the original peptide. In contrast, substitutions of an amino acid in a peptide by another amino acid in a different group are generally more likely to alter the characteristics of the original peptide. The present invention also contemplates substitution with non-naturally occurring amino acids.

In any embodiment disclosed herein, the compound may be of Formula I:

NH2ZZXCXXXXXXCXXXXXCCXXXXCXXXCXZZYYZZNH2;

Wherein each Z is either absent, any amino acid or any alkyne or azide containing amino acid derivative excluding cysteine, and each X is any amino acid, or any alkyne or azide containing amino acid derivative excluding cysteine, and each Y is a polar or non-polar or negatively charged amino acid excluding cysteine, and each C is cysteine, and the C-terminus is amidated. Examples of amino acids include: Ala(A), Arg(R), Asn(N) , Asp(D), Gln(Q, Glu(E), Gly(G), His(H) , lle(l), Leu(L), Lys(K), Met(M) , Phe(F) , Pro(P), Ser (S), Thr(T), Trp(W), Tyr(Y), Val(V). Examples of alkyne containing amino acid derivatives include: Homopropargyl- glycine (Hpg), 2-Propargyl-glycine (Pra), 4-Propargyl-proline, 6-Propargyl-lysine, 3-Propargyl- alanine. Examples of azide containing amino acid derivatives include: 3-Azido-alanine, 4- Azido-homoalanine, 4-Azido-phenylalanine, 5-Azido-ornithine, 6-Azido-lysine, 2-Azido- valine, 2-Azido-3-phenylpropionic acid, 2-Azido-aspertane, 3-Azido-alanine. Examples of polar amino acids include: Ser(S), Thr (T), Tyr (Y), Asn (N), Gln(Q). Examples of non-polar amino acids include: Gly(G), Ala(A), Val(V), Pro(P), Leu(L), lle(l), Met(M), Trp(W), Phe(F). Examples of negatively charged amino acids include: Asp(D), Glu(E).

The pharmacokinetics of any of the disclosed peptides can be determined after administration of the peptide through different routes of administration. For example, different routes of administration of the disclosed peptides are intravenous, subcutaneous, intramuscular, rectal, aerosol, ocular, lung, transdermal, vagina, eye, nasal, oral, sublingual, inhalation, skin, arachnoid membrane, intranasal, intraarticular, peritoneal, buccal, synovial, or topical administration. The peptides of the present disclosure can be analyzed using tracking agents such as radiolabels or fluorophores. For example, a radiolabeled peptide of the present disclosure can be administered through various routes of administration and peptide concentration or dose recovery can be measured in various biological samples such as plasma, urine, stool, any organ, skin, muscle, and other tissues; and this can be measured by HPLC, radio HPLC, radio TLC, gamma counter or liquid scintillation counting. As another example, a fluorophore labeled peptide of the present disclosure can be measured by fluorescence detection techniques (TECAN quantification, flow cytometry, I VIS) or a series of methods, including.

As used herein, a "radiolysis protection agent" refers to any molecule, compound or composition that may be added to a radionuclide-labeled complex or molecule to decrease the rate of breakdown of the radiolabeled complex or molecule by radiolysis. Any known radiolysis protection agent, including but not limited to ethanol, gentisic acid, or ascorbic acid, may be used.

The peptides used are conveniently synthesized on an automated peptide synthesizer using a solid-phase support and standard techniques of repetitive orthogonal deprotection and coupling. Free amino groups in the peptide, that are to be used later for conjugation of chelating moieties or other agents, are advantageously blocked with standard protecting groups such as a Boc group, while N-terminal residues may be acetylated to increase serum stability. Such protecting groups are well known to the skilled artisan. See Greene and Wuts Protective Groups in Organic Synthesis, 1999 (John Wiley and Sons, N.Y.). Peptides are advantageously cleaved from the resins to generate the corresponding C-terminal amides, in order to inhibit in vivo carboxypeptidase activity. Exemplary structures of use and methods of peptide synthesis are disclosed in the Examples below. Chelating moieties may be conjugated to peptides using bifunctional chelating moieties.

Some embodiments may involve substitution of one or more D-amino acids for the corresponding L-amino acids. Peptides comprising D-amino acid residues are more resistant to peptidase activity than L-amino acid comprising peptides. Such substitutions may be readily performed using standard amino acid synthesizers.

Non-limiting examples of diagnostic agents may include a radionuclide such as 11C, 150, 18F, 32P, 51Mn, 52Fe, 52Mn, 55Co, 62Cu, 64Cu, 67Cu, 67Ga, 68Ga, 72As, 75Br, 76Br, 82mRb, 83Sr, 86Y, 89Zr, 90Y, 94mTc, 94Tc, 99mTc, lllln, 1201, 1231, 1241, 1251, 13N, 1311, 154-158Gd, 177Lu, 186Re, 188Re, or other gamma-, beta-, or positron-emitters. Other non limiting examples of diagnostic agents may include a paramagnetic ion such as chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III) or erbium (III). Other non-limiting examples of diagnostic agents may include a metal contrast agent such as lanthanum (III), gold (III), lead (II) or bismuth (III).

Other non-limiting examples of diagnostic agents may include a radiopaque diagnostic agent selected from iodine compounds, barium compounds, gallium compounds, and thallium compounds.

Other non-limiting examples of diagnostic agents may include a fluorescent label such as fluorescein isothiocyanate, rhodamine, phycoerytherin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine. Other non-limiting examples of diagnostic agents may include a chemiluminescent labels of use may include luminol, isoluminol, an aromatic acridinium ester, an imidazole, an acridinium salt or an oxalate ester.

Non-limiting examples of radionuclide therapy agents may include a radionuclide such as 67Cu, 68Ga, 77Br, 89Sr, 90Y, 105Rh, lllln, 1231, 1251, 1311, 149Pm, 166Ho, 177Lu, 186Re, 188Re, 199Au, 211At, 212Bi, 212Pb, 213Bi, 223Ra, 224Ra, 225Ac, 227Th, or other alpha- beta-, or auger-emitters.

Radiolabeled peptides of the present invention can be useful as PET probes. A PET probes would provide a quantitative readout of pain, improving on the current, often qualitative assessments. This can be used, for example, as a way to way to image pain before the assignment of treatment (e.g. opioid medications, surgery), or as an evaluation of ongoing treatment, or as a tool to monitor newly assigned treatment. A PET probe would also provide quantitative readout of the physiological condition of the peripheral nerves. This can be used, for example, as a way to locate nerve injury, or as a way to monitor other treatments (e.g. chemotherapy, surgery) that are capable of impairing the peripheral nerves. Radiolabeled peptides of the present invention are also useful as molecular targeting agents for radionuclide therapy.

A compound can consist of a fluorophore conjugated to a peptide of SEQ ID NOs. 1- 514. Non-limiting examples of fluorophores are: Squaranines, rhodamines, cyanines, AF350, AF405, AF430, AF488, AF514, AF532, AF546, AF555, AF568, AF594, BDP503, BDP550, BDP568, BDP570, BDP589, BDP591, BDP-FL, BDP-R6G, BDP-TMR, CY3, CY3.5, Fluorescein, JF549, JF585. Non-limiting examples of IR fluorophores are: BODIPY derivatives, rhodamine derivatives, squaranine derivatives, cyanine derivatives, AF610, AF633, AF635, AF647, AF660, AF680, AF700, AF750, AF790, BDP650, BDP665, BDPTR, CY5, CY5.5, CY7, CY7.5, DY675, DY676, DY677, DY678, DY679P1, DY680, DY681, DY682, DY684, ICG, IR650, IR680, IR750, IR800, JF635, JF646, JF669.

Fluorophores of the present invention can be attached to peptides at the following amino acids: lysine (Lys), 2-Propargyl-glycine (Pra), homopropargyl-glycine (Hpg), 4- Propargyl-proline, 6-Propargyl-lysine, 3-Propargyl-alanine, 3-Azido-alanine, 4-Azido- homoalanine, 4-Azido-phenylalanine, 5-Azido-ornithine, 6-Azido-lysine, 2-Azido-valine, 2- Azido-3-phenylpropionic acid, 2-Azido-aspertane, 3-Azido-alanine.

The fluorophore conjugated peptides can be useful as fluorescent probes. A fluorescent probe is primarily for imaging peripheral nerves during surgery in order to avoid iatrogenic nerve injuries resulting in pain, paresthesia, abnormal sensation or loss of function. They can be used, for example, in orthopedic procedures such as osteosynthesis, osteotomy, or repair of ruptured ligaments. As another example, this method can be used in minor surgeries such as lymph node biopsy, ganglion cyst removal, or lipoma removal. As another example, this method can be used in endoscopic or open hand surgeries, or other hand surgeries such as index finger release. As another example, this method can be used in cancer related surgeries such as thyroidectomy, prostatectomy or mastectomy. As another example, this method can be used in varicose vein operations. As another example, this method can be used in hernia repair.

The labeled peptides may be formulated to obtain compositions that include one or more pharmaceutically suitable excipients, one or more additional ingredients, or some combination of these. These can be accomplished by known methods to prepare pharmaceutically useful dosages, whereby the active ingredients (i.e., the labeled peptides) are combined in a mixture with one or more pharmaceutically suitable excipients. Sterile phosphate-buffered saline is one example of a pharmaceutically suitable excipient. Other suitable excipients are well known to those in the art.

A preferred route for administration of the compositions described herein is parenteral injection. Injection may be intravenous, intraarterial, intralymphatic, intrathecal, subcutaneous or intracavitary (i.e., pa rente rally). In parenteral administration, the compositions will be formulated in a unit dosage injectable form such as a solution, suspension or emulsion, in association with a pharmaceutically acceptable excipient. Such excipients are inherently nontoxic and nontherapeutic. Examples of such excipients are saline, Ringer's solution, dextrose solution and Hank's solution. Nonaqueous excipients such as fixed oils and ethyl oleate may also be used. A preferred excipient is 5% dextrose in saline. The excipient may contain minor amounts of additives such as substances that enhance isotonicity and chemical stability, including buffers and preservatives.

Another preferred route for administration of the compositions described herein is by oral administration. Compositions for oral administration will be formulated in dosage forms suitable for oral administration, including: pills; lozenges; capsules; powders; films; oral dissolving tablets; and liquids. Dosage forms for oral administration may be immediate release, extended release, or modified release. Dosage forms for oral administration may include enteric coatings, extended release coatings, modified release coating, or no coating. Oral dosage forms may include any acceptable excipients, including: bulking agents; dissolution enhancers; wetting agents; surfactants; flavoring agents; coatings; and preservatives. A preferred excipient is mannitol.

Formulated compositions comprising labeled targeting peptides can be used for intravenous administration via, for example, bolus injection or continuous infusion. Compositions for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. Compositions can also take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the compositions can be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

The compositions may be administered in solution. The pH of the solution should be in the range of pH 5 to 9.5, preferably pH 6.5 to 7.5. The formulation thereof should be in a solution having a suitable pharmaceutically acceptable buffer such as phosphate, TRIS (hydroxymethyl) aminomethane-HCI or citrate and the like. In certain preferred embodiments, the buffer is potassium biphthalate (KHP), which may act as a transfer ligand to facilitate 18F-labeling. Buffer concentrations should be in the range of 1 to 100 mM. The formulated solution may also contain a salt, such as sodium chloride or potassium chloride in a concentration of 50 to 150 mM. An effective amount of a stabilizing agent such as glycerol, albumin, a globulin, a detergent, a gelatin, a protamine or a salt of protamine may also be included. The compositions may be administered to a mammal subcutaneously, intravenously, intramuscularly or by other parenteral routes. Moreover, the administration may be by continuous infusion or by single or multiple boluses.

In general, the dosage of 18F or other radiolabel to administer to a human subject will vary depending upon such factors as the patient's age, weight, height, sex, general medical condition and previous medical history. Preferably, a saturating dose of the labeled molecule is administered to a patient. For administration of radiolabeled molecules, the dosage may be measured by millicuries. A typical range for imaging studies would be 5 to 10 mCi.

Various embodiments of the claimed methods and/or compositions may concern one or more 18F- or other radiolabeled peptides to be administered to a subject. Administration may occur by any route known in the art, including but not limited to oral, nasal, buccal, inhalational, rectal, vaginal, topical, orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal, intraarterial, intrathecal or intravenous injection.

In certain embodiments, the standard peptide bond linkage may be replaced by one or more alternative linking groups, such as CH2-NH, CH2-S, CH2-CH2, CH=CH, CO-CH2, CHOH-CH2 and the like. Peptide mimetics may exhibit enhanced stability and/or absorption in vivo compared to their peptide analogs.

Methods of diagnostic imaging with labeled peptides are well-known. For example, in the 2D technique of scintigraphy, peptide ligands are labeled with a gamma-emitting radioisotope and introduced into a patient. A gamma camera is used to detect the location and distribution of gamma-emitting radioisotopes. 3D techniques include the use of gamma-emitting radionuclides (SPECT isotopes), with gamma energies typically ranging from 50-400 keV, such as 67Ga, 99mTc, lllln, 1231, 1311, and 201TI. Such radionuclides may be imaged by well-known SPECT scanning techniques. 3D techniques also include the use of positron-emitting radionuclides (PET isotopes), with a gamma energy of 511 keV through annihilation, such as 18F, 68Ga, 64Cu, 89Zr, and 1241. Such radionuclides may be imaged by well-known PET scanning techniques.

In an aspect, a composition is provided that includes a compound of any aspect or embodiment disclosed herein and a pharmaceutically acceptable carrier or one or more excipients, fillers or agents (collectively referred to hereafter as "pharmaceutically acceptable carrier" unless otherwise indicated and/or specified). In a related aspect, a medicament is provided that includes a compound of any aspect or embodiment disclosed herein. In a related aspect, a pharmaceutical composition is provided that includes (i) an effective amount of a compound of any aspect or embodiment disclosed herein, and (ii) a pharmaceutically acceptable carrier. For ease of reference, the compositions, medicaments, and pharmaceutical compositions of the present technology may collectively be referred to herein as "compositions." In further related aspects, the present technology provides methods including a compound of any embodiment disclosed herein and/or a composition of any embodiment disclosed herein as well as uses of a compound of any embodiment disclosed herein and/or a composition of any embodiment disclosed herein. Such methods and uses may include an effective amount of a compound of any embodiment disclosed herein.

In any aspect or embodiment disclosed herein, the effective amount may be determined in relation to a subject or group of subjects. "Effective amount" refers to the amount of a compound or composition required to produce a desired effect. One non limiting example of an effective amount includes amounts or dosages that yield acceptable toxicity and bioavailability levels for therapeutic (pharmaceutical) use including, but not limited to, the treatment of pain or for the imaging of peripheral neurons. In any aspect or embodiment disclosed herein (collectively referred to herein as "any embodiment herein," "any embodiment disclosed herein," or the like) of the compositions, pharmaceutical compositions, and methods including compounds of the present technology, the effective amount may be an imaging-effective amount of the compound for imaging peripheral neurons in a subject. An "imaging-effective amount" refers to the amount of a compound or composition required to produce a desired imaging effect, such as a quantity of a compound of the present technology necessary to be detected by the detection method chosen. For example, an effective amount of a compound of the present technology includes an amount sufficient to enable detection of binding of the compound to peripheral neurons. Another example of an effective amount includes amounts or dosages that are capable of providing a fluorescence emission (above background) in peripheral neurons in a subject, such as, for example, statistically significant emission above background. As used herein, a "subject" or "patient" is a mammal, such as a cat, dog, rodent or primate. Typically the subject is a human. In any embodiment herein of the compositions, pharmaceutical compositions, and methods including proteins of the present technology, the effective amount may be an amount that treats pain in a subject. Such treatment may include a statistically significant reduction of perceived pain over such performance absent administration of the compound; such an increase may be a statistically significant increase over such performance as compared to administration of an equivalent amount of a comparative standard in terms of moles. By way of example, the effective amount of any embodiment herein including proteins of the present technology may be from about 0.01 ng to about 1,000 mg of the compound per gram of the composition, preferably from about 0.01 mg to about 200 mg of the compound per gram of the composition, and further preferably from about 0.1 mg to about 10 mg of the compound per gram of the composition. By further way of example, for PET imaging, an effective amount may include the synthesis of between 50 and 5000 ug, and preferably between 200 and 500 ug, and may include administering preferentially between 2 and 100 mol, and more preferentially between 5 and 30 nmol. By further way of example, for, an effective amount for PET imaging or radiotherapy may include the synthesis of between 50 and 5000 ug, and preferably between 200 and 500 ug, and may include administering preferentially between 2 and 100 mol, and more preferentially between 5 and 30 nmol.

The pharmaceutical composition of any embodiment disclosed herein may be packaged in unit dosage form. The unit dosage form is effective in treating pain (when proteins of the present technology are included) or is effective in imaging peripheral neurons (when compounds of the present technology are included). Generally, a unit dosage including a compound of the present technology will vary depending on patient considerations. Such considerations include, for example, age, protocol, condition, sex, extent of disease, contraindications, concomitant therapies and the like. An exemplary unit dosage based on these considerations may also be adjusted or modified by a physician skilled in the art. For example, a unit dosage for a patient comprising a compound of the present technology may vary from 1 x 10-4 g/kg to 1 g/kg, preferably, 1 x 10-3 g/kg to 1.0 g/kg. Dosage of a compound of the present technology may also vary from 0.01 mg/kg to 100 mg/kg or, preferably, from 0.1 mg/kg to 10 mg/kg. Suitable unit dosage forms, include, but are not limited to parenteral solutions, oral solutions, powders, tablets, pills, gelcaps, capsules, lozenges, suppositories, patches, nasal sprays, injectables, implantable sustained- release formulations, mucoadherent films, topical varnishes, lipid complexes, liquids, etc.

The compositions of the present technology may be prepared by mixing one or more compounds of any embodiment disclosed herein of the present technology with one or more pharmaceutically acceptable carriers in order to provide a pharmaceutical composition useful to prevent and/or treat pain (when proteins of the present technology are included) or useful in imaging peripheral neurons (when compounds of the present technology are included). Such compositions may be in the form of, for example, granules, powders, tablets, capsules, syrup, suppositories, injections, emulsions, elixirs, suspensions or solutions. The instant compositions may be formulated for various routes of administration, for example, by oral, parenteral, topical, rectal, nasal, vaginal administration, or via implanted reservoir. Parenteral or systemic administration includes, but is not limited to, subcutaneous, intravenous, intraperitoneal, and intramuscular injections. The following dosage forms are given by way of example and should not be construed as limiting the present technology. The compounds of the present invention may be used in combination with one or more other compounds including but not limited to antihistamines, NSAIDs, anti-inflammatories, opioids, anti-coagulants, anti-neoplastic agents, antibody therapy, tyrosine kinase inhibitors, DNA methylation agents, angiogenesis inhibitors, hormonal therapy, immunotherapy, antimetabolites, topoisomerase inhibitors, cross-linking agents, photosensitizers, receptor antagonists, enzyme inhibitors, microtubule blockers, or DMARDs.

For oral, buccal, and sublingual administration, powders, suspensions, granules, tablets, pills, capsules, gel caps, and caplets are acceptable as solid dosage forms. These can be prepared, for example, by mixing one or more compounds of the instant present technology with at least one additive such as a starch or other additive. Suitable additives include sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates, chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins, collagens, casein, albumin, synthetic or semi-synthetic polymers or glycerides. Optionally, oral dosage forms can contain other ingredients to aid in administration, such as an inactive diluent, or lubricants such as magnesium stearate, or preservatives such as para ben or sorbic acid, or anti oxidants such as ascorbic acid, tocopherol or cysteine, a disintegrating agent, enteric coatings, controlled release coatings, binders, thickeners, buffers, sweeteners, flavoring agents, perfuming agents, or a combination of any two or more thereof. Tablets and pills may be further treated with suitable coating materials known in the art, including immediate release, extended release and modified release coatings.

Liquid dosage forms for oral administration may be in the form of pharmaceutically acceptable emulsions, syrups, elixirs, suspensions, and solutions, which may contain an inactive diluent, such as water. Pharmaceutical compositions may be prepared as liquid suspensions or solutions using a sterile liquid, such as, but not limited to, an oil, water, an alcohol, and combinations of these. Pharmaceutically suitable surfactants, stabilizers, antioxidants, suspending agents, emulsifying agents, buffers, pH modifiers, or a combination of any two or more thereof, may be added for oral or parenteral administration.

Injectable dosage forms generally include aqueous suspensions or oil suspensions which may be prepared using a suitable dispersant or wetting agent and a suspending agent. Injectable forms may be in solution phase or in the form of a suspension, which is prepared with a solvent or diluent. Acceptable solvents or vehicles include sterilized water, Ringer's solution, or an isotonic aqueous saline solution. Additionally or alternatively, sterile oils may be employed as solvents or suspending agents. Typically, the oil or fatty acid is non volatile, including natural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the composition may be a powder suitable for reconstitution with an appropriate solution as described above. Examples of these include, but are not limited to, freeze dried, rotary dried or spray dried powders, amorphous powders, granules, precipitates, or particulates. For injection, the formulations may optionally contain stabilizers, pH modifiers, buffers, surfactants, bioavailability modifiers, and combinations of any two or more of these.

Compounds of the present technology may be administered to the lungs by inhalation through the nose or mouth. Suitable compositions for inhalation include solutions, sprays, dry powders, or aerosols containing any appropriate solvents and optionally other compounds such as, but not limited to, stabilizers, antimicrobial agents, antioxidants, pH modifiers, surfactants, bioavailability modifiers and combinations of these. The carriers and stabilizers vary with the requirements of the particular compound, but typically include nonionic surfactants (Tweens, Pluronics, or polyethylene glycol), innocuous proteins like serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aqueous and non-aqueous (e.g., in a fluorocarbon propellant) aerosols may be used for delivery of compounds of the present technology by inhalation.

Dosage forms for the topical (including buccal and sublingual) or transdermal administration of compounds of the present technology include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, and patches. The active component may be mixed under sterile conditions with a pharmaceutically acceptable carrier and/or excipient, and with any preservatives, or buffers, which may be required. Powders and sprays can be prepared, for example, with excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. The ointments, pastes, creams and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof. Absorption enhancers can also be used to increase the flux of the compounds of the present technology across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane (e.g., as part of a transdermal patch) or dispersing the compound in a polymer matrix or gel.

Besides those representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the instant present technology. Such excipients and carriers are described, for example, in "Remington's Pharmaceutical Sciences" Mack Pub. Co., New Jersey (1991), and "Handbook of Pharmaceutical Excipients" by Raymond Rowe. Pharmaceutical Press, London, UK (2009), each of which is incorporated herein by reference.

The compositions (e.g., pharmaceutical compositions) of the present technology may be designed to be short-acting, fast-releasing, long-acting, and sustained-releasing as described below. Thus, the compositions may also be formulated for controlled release or for slow release.

The compositions of the present technology may also include, for example, micelles or liposomes, or some other encapsulated form, or may be administered in an extended release form to provide a prolonged storage and/or delivery effect. Therefore, the compositions may be compressed into pellets or cylinders and implanted intramuscularly or subcutaneously as depot injections or as implants such as stents. Such implants may employ known inert materials such as silicones and biodegradable polymers.

In one aspect, the present invention provides for a method for the treatment of pain by administering a compound of SEQ ID NOs 1-514 and Formula I. This method can be used to treat, for example, acute pain, chronic pain, nociceptive pain, inflammatory pain, neuropathic pain, and functional pain. This method can be used to treat pain arising from conditions including, but not limited to, diabetic neuropathy, fibromyalgia, post-operative pain, migraine, other neuropathy, headache, arthritis pain, mechanical pain (e.g. back pain), gout, rheumatoid arthritis, neuropathy, radicular pain, trigeminal neuralgia, irritable bowel syndrome, cancer related-pain, repetitive strain injury pain, burn pain, animal bite or sting pain, sciatica pain, multiple sclerosis related pain, lupus related pain, ankylosing spondylitis related pain, unspecified connective tissue disorder pain, phantom limb pain, ulcer related pain, myelopathy related pain, claudication related pain, STI related pain, muscular dystrophy related pain, analgesic rebound, chemical substance withdrawal related pain, radiculopathy related pain, anatomical related pain (e.g., os trignum syndrome), ocular pain (e.g., uveitis, iritis, retinopathy), food poisoning related pain, rhabdomyolysis related pain, poisoning related pain (e.g. mycotoxin exposure related pain), nutritional deficiency related pain (e.g., anemia related pain), menstrual pain, dystonia related pain, and spasticity related pain.

In one aspect, the present invention provides a method for the treatment of itching by administering a compound of SEQ ID NOs 1-514 and Formula I. This method can be used to treat, for example, urticaria, dermatological irregularities, rashes, eczema, chicken pox, surgical healing and the like.

In one aspect, the present invention provides a method for the treatment of cancer by administering a compound of SEQ ID NOs 1-514 and Formula I. This method can be used to treat, for example, endometrial cancer, gastric cancer, lung cancer, ovarian cancer, and prostate cancer and the like. Treatment may be achieved by inhibition of NaV1.7 or by using NaV1.7 targeting to bring a radionuclide to the cancer for radionuclide therapy. In a preferred embodiment, the NaV1.7 inhibiting peptide compound of SEQ ID NOs 1-514 and Formula I includes a radioactive atom that is bound to the peptide either via a chemical bond or via chelation or other inter-molecular forces, including 177Lu.

In one aspect, the present invention provides a method for imaging pain by administering a compound of SEQ ID NOs 1-514 and Formula I. This method can be used to locate and quantify pain to enable more precise pain management. In a preferred embodiment, the NaV1.7 inhibiting peptide compound of SEQ ID NOs 1-514 and Formula I includes a radioactive atom that is bound to the peptide either via a chemical bond or via chelation or other inter-molecular forces, including 18F.

In one aspect, the present invention provides a method for imaging peripheral neurons by administering a compound of SEQ ID NOs 1-514 and Formula I. This method can be used for to monitor the location of nerves during surgery so as to avoid them if possible. Examples of such surgeries include, but are not limited to: horacotomy, breast surgery, cholecystectomy, inguinal herniorrhaphy, and radical prostatectomy. In a preferred embodiment, the NaV1.7 inhibiting peptide compound of SEQ ID NOs 1-514 and Formula I contains a fluorophore, including BODIPY.

In one aspect, the present invention provides methods to manage conditions in either communicative or non-communicative patients, including human and veterinary patients. This method can be used to detect over-expression of receptors associated with the conditions being treated, including pain. The detection of an over-expression of such receptors can be used to diagnose untreated pain in non-communicative patients or patients that cannot identify the location of said pain.

Examples

Example 1: Production of SEQ. ID NO 1-514

The peptides of SEQ ID NO 1-514 will be synthesized on an automated peptide synthesizer using a solid-phase support and standard techniques of repetitive orthogonal deprotection and coupling. Free amino groups in the peptide, that will be used later for conjugation of chelating moieties or other agents, will be advantageously blocked with standard protecting groups such as a Boc group, while N-terminal residues may be acetylated to increase serum stability. Peptides will be cleaved from the resins to generate the corresponding C-terminal amides, in order to inhibit in vivo carboxypeptidase activity. Alternatively, the peptides of SEQ ID NO 1-514 will be synthesized using recombinant periplasmic expression in bacterial hosts, including E. coli.

The examples herein are provided to illustrate advantages of the present technology and to further assist a person of ordinary skill in the art with preparing and/or using the compounds of the present technology. The examples should in no way be construed as limiting the scope of the present technology.

Claims

Claims

Claim 1. A peptide comprising SEQ ID NO: 2

( Y CQKWLWT CD SERPCCEGL V CRLW CKIN-NEE), or a substituted variant thereof, or a pharmaceutically acceptable salt and/or solvate thereof.