TREA

NERVE TARGETING PEPTIDE CONJUGATES AND METHODS OF USE

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Information

  • Patent Application
  • 20240207422
  • Publication Number
    20240207422
  • Date Filed
    December 06, 2023
    a year ago
  • Date Published
    June 27, 2024
    7 months ago
Information
Abstract
Nerve targeting peptides, including those comprising the amino acid sequence of any one of SEQ ID NOS:1-557, Formulas (I)-(XLIV), and Subformulas (Ia)-(XLIVa); conjugates comprising such nerve targeting peptides and a cargo molecule; compositions comprising such peptides and conjugates; and their use in methods of fluorescent guided surgery and other applications.
Description
REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

The contents of the electronic sequence listing (409_SeqListing.xml; Size: 504,874 bytes; and Date of Creation: Dec. 5, 2023) is herein incorporated by reference in its entirety.


FIELD

The present invention relates to nerve targeting peptides and conjugates thereof and their use in intraoperative nerve visualization and other targeted surgical and medical applications.


BACKGROUND

Preserving human neurons and human nerves is an important goal of surgical procedures; for example, accidental transection of nerves can lead to significant morbidity, such as chronic pain and loss of function. Nerves are typically identified by their elongated whitish appearance and relationship to nearby structures or by electrophysiological studies. However, such nerve identification can be difficult. Nerve structures can be obscured or distorted in complex surgical settings, such as those involving trauma, tumors, inflammation, or infection. Nerve visibility can be further restricted in minimally invasive procedures, such an endoscopy, which rely on small incisions with limited access. Indeed, inadvertent nerve damage is a leading complication associated with many surgeries using both open and minimal access procedures.


These observations highlight the need for methods and tools to identify neurons or nerves reliably and efficiently. Electromyography (EMG) offers one approach to neuron or nerve identification prior to direct exposure during surgery or confirmation of neuron or nerve identity in instances of uncertainty. A disadvantage of this technique, however, is the lack of visual feedback to the operating surgeon. Even if a nerve has been identified in one location, either through accidental or purposeful stimulation, there is no visual guidance to the operating surgeon as to how far away from the stimulation site the nerve lies or the direction of travel the nerve takes away from the stimulation site. Furthermore, EMG only traces motor pathways, not sensory fibers. EMG can fail if neuron or nerve conduction or neuromuscular transmission is temporarily blocked anywhere distal to the recording site. Such blockade easily occurs due to neuron or nerve compression, trauma, local anesthetics, or neuromuscular blockers.


Other approaches include neuron or nerve labeling, which primarily depends on retrograde or anterograde tracing of individually identified axonal tracts via the use of fluorescent dyes. However, methods of labeling neurons or nerves by locally applied fluorescent tracers can be limited. First, depending on where the dye has been injected, this technique can label only one neuron or nerve fiber tract at a time. Second, this technique may result in only limited labeling of fluorescent dyes along the axonal tracts because retrograde axonal tracers typically accumulate in the neural cell body. Third, retrograde transport is relatively slow (on the order of millimeters per day) and therefore takes a long time to label human neuron or nerves, which are often longer than a meter, such as in the case of the sciatic neuron or nerve and its arborizations. Fourth, the application of fluorescent dyes to innervation targets, such as direct intramuscular injections to label a motor neuron or nerves, is typically messy with a variable amount of the tracer dye remaining at the injection site. As dissection of neurons or nerves depends on accurate visualization of adjacent structures prior to encountering them, a surgical site that is contaminated with fluorescent dyes would not be desirable. Finally, the direct injection of the fluorescent dye itself may be damaging to the target organs or neuron or nerve of interest, either by mechanical damage or by the very high local concentration of dye and vehicle at the injection site.


Accordingly, there is a need in the art to identify peptides capable of selective binding to human nerves and neurons, to facilitate surgical procedures and human nerve protection. The present disclosure provides peptide sequences that selectively bind to human nerves or neurons, as well as methods of using those peptides, for example in conjunction with selected cargo molecules to allow fluorescent visualization of nerves during surgery, or to target drugs and other agents to nerves.


BRIEF SUMMARY

In one aspect, the present disclosure provides a nerve targeting peptide conjugate comprising: a peptide consisting or comprising the amino acid sequence of any one of SEQ ID NOS: 1-557; and a cargo molecule. In one aspect, the present disclosure provides a nerve targeting peptide conjugate comprising: a peptide consisting or comprising the amino acid sequence of any one of Formulas I-XLIV, as described herein, including Subformulas Ia-XLIVa; and a cargo molecule.


In some embodiments, the cargo molecule comprises a drug, fluorescent moiety, or photosensitizing agent.


In some embodiments, the cargo molecule is joined to the N-terminus or C-terminus of the peptide.


In some embodiments, the cargo molecule is joined to the peptide via a linker.


In some embodiments, the linker is a straight or branched-chain carbon linker, heterocyclic carbon linker, amino acid linker, lipophilic residue, peptide linker, peptide nucleic acid linker, hydrazone linker, SPDB disulfide, sulfo-SPDB, maleimidomethyl cyclohexane-1-carboxylate (MCC), aminohexanoic acid linker, polyether linker, or polyethylene glycol linker.


In some embodiments, the cargo molecule comprises a fluorescent moiety.


In some embodiments, the fluorescent moiety is a fluorescent protein, a fluorescent peptide, a fluorophore, or any combination thereof.


In some embodiments, the fluorescent moiety is selected from the group consisting of: a xanthene; a bimane; a coumarin; an aromatic amines; a benzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylene pyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare-earth metal chelate complexes; and derivatives thereof.


In some embodiments, the fluorescent moiety is selected from the group consisting of: 5-carboxyfluorescein; fluorescein-5-isothiocyanate; 6-carboxyfluorescein; tetramethylrhodamine-6-isothiocyanate; 5-carboxytetramethylrhodamine; 5-carboxy rhodol derivatives; tetramethyl and tetraethyl rhodamine; diphenyldimethyl and diphenyldiethyl rhodamine; dinaphthyl rhodamine; rhodamine 101 sulfonyl chloride; Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7, indocyanine green, IR800CW, cyan fluorescent protein (CFP), EGFP, 6-FAM, FAM, fluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluorescein zwitterion, carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate, carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,5′(6′)-carboxyfluorescein, fluorescein glutathione, or any combination thereof.


In some embodiments, the cargo molecule comprises a photosensitizing agent.


In some embodiments, the photosensitizing agent is selected from the group consisting of: a porphyrin, chlorin, and dye.


In some embodiments, the photosensitizing agent is selected from the group consisting of: porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid, Lutetium texaphyrin, metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazine, metallo-naphthalocyanines, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine derivative, Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF), IR700DX, naphthalocyanine, and porphyrin derivatives.


In some embodiments, the cargo molecule comprises a drug.


In some embodiments, the drug is selected from the group consisting of: an antihistamine, a GABA receptor modulator, a neurotransmitter reuptake inhibitor, a local anesthetic, an anticholinergic, a sodium channel blocker, a calcium channel blocker, a thyrotropin-releasing hormone, a γ-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDA receptor agonist or antagonist, an mGlu receptor agonist or antagonist, a growth factor, an antiemetic agent, a corticosteroid; a nonsteroidal anti-inflammatory drug (NSAID); an anti-epileptic agent; a neurotropic agent; a cytotoxic agent; an antioxidant, an iron chelator, a mitochondrial modulator, a sirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase (NOS) modulator, a potassium channel agonist or antagonist, a purigenic receptor agonist or antagonist, or any combination thereof.


In some embodiments, the drug is selected from the group consisting of: benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine; prilocaine; propxycaine; proparacaine; tetracaine; tocainide; and trimecaine; methotrexate; cyclophosphamide; thalidomide; paclitaxel; pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; platonin; procarbazine; raltitrexed; rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA; tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate; tris(2-chloroethyl)amine; troxacitabine; uracil mustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; carbamazepine; oxcarbazepine; phenytein; valproic acid; sodium valproate; cinnarizine; flunarizine; nimodipine; brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived neurotrophic factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-like growth factor (IGF); prednisone; prednisolone; dexamethasone; gabapentin; or any combination thereof.


In another aspect, the present disclosure provides a pharmaceutical composition comprising a nerve targeting peptide conjugate described herein and a pharmaceutically acceptable carrier.


In yet another aspect, the present disclosure provides a method of delivering a cargo molecule to a neuron or nerve comprising contacting the neuron or nerve with a nerve targeting peptide conjugate of the present disclosure.


In yet another aspect, the present disclosure provides a method of identifying a neuron or nerve comprising contacting the neuron or nerve with a nerve targeting peptide conjugate of the present disclosure.


In yet another aspect, the present disclosure provides a method of delivering a drug to a neuron or nerve comprising contacting the neuron or nerve with a nerve targeting peptide conjugate of the present disclosure.


In another aspect, the present disclosure provides a method of delivering a photosensitizing agent to a neuron or nerve comprising contacting the neuron or nerve with a nerve targeting peptide conjugate of the present disclosure.





BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.



FIGS. 1A-1B are Venn diagrams depicting: (FIG. 1A) the overlap of peptide sequence subsets identified from phage display screens directed to two independent human muscle samples (muscle 2 and muscle 3) and three independent human nerve samples (nerve2, nerve3, and plated_nerve2); and (FIG. 1B) the source of the representative sample of nerve-specific peptides, namely those comprising peptides unique to each nerve subset (highlighted in gray), peptides in common between two nerve subsets (highlighted in yellow), and peptides found in all three nerve subsets (highlighted in green).



FIGS. 2A-2B are graphical depictions of the most prevalent amino acids at each position of the representative sample of nerve-binding peptides identified in Example 1, based on (FIG. 2A) unweighted or (FIG. 2B) weighted probabilities.



FIG. 3 shows the alignment of peptide sequences comprising cluster 1, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The ID number of each peptide is indicated, and the “majority rule consensus” residues at each position are highlighted to identify patterns reflected in corresponding Formula I and Subformula Ia.



FIG. 4 shows the alignment of peptide sequences comprising cluster 2, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns reflected in corresponding Formula II and Subformula IIa.



FIG. 5 shows the alignment of peptide sequences comprising cluster 3, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns reflected in corresponding Formula III and Subformula IIIa.



FIG. 6 shows the alignment of peptide sequences comprising cluster 4, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns reflected in corresponding Formula IV and Subformula IVa.



FIG. 7 shows the alignment of peptide sequences comprising cluster 5, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns reflected in corresponding Formula V and Subformula Va.



FIG. 8 shows the alignment of peptide sequences comprising cluster 6, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula VI and Subformula VIa.



FIG. 9 shows the alignment of peptide sequences comprising cluster 7, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula VII and Subformula VIIa.



FIG. 10 shows the alignment of peptide sequences comprising cluster 8, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula VIII and Subformula VIIIa.



FIG. 11 shows the alignment of peptide sequences comprising cluster 9, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula IX and Subformula IXa.



FIG. 12 shows the alignment of peptide sequences comprising cluster 10, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula X and Subformula Xa.



FIG. 13 shows the alignment of peptide sequences comprising cluster 11, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XI and Subformula XIa.



FIG. 14 shows the alignment of peptide sequences comprising cluster 12, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XII and Subformula XIIa.



FIG. 15 shows the alignment of peptide sequences comprising cluster 13, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XIII and Subformula XIIIa.



FIG. 16 shows the alignment of peptide sequences comprising cluster 14, based on Clustal Omega analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XIV and Subformula XIVa.



FIG. 17 shows the alignment of peptide sequences comprising cluster 15, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XV and Subformula XVa.



FIG. 18 shows the alignment of peptide sequences comprising cluster 16, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XVI and Subformula XVIa.



FIG. 19 shows the alignment of peptide sequences comprising cluster 17, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XVII and Subformula XVIIa.



FIG. 20 shows the alignment of peptide sequences comprising cluster 18, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XVIII and Subformula XVIIIa.



FIG. 21 shows the alignment of peptide sequences comprising cluster 19, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XIX and Subformula XIXa.



FIG. 22 shows the alignment of peptide sequences comprising cluster 20, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XX and Subformula XXa.



FIG. 23 shows the alignment of peptide sequences comprising cluster 21, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXI and Subformula XXIa.



FIG. 24 shows the alignment of peptide sequences comprising cluster 22, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXII and Subformula XXIIa.



FIG. 25 shows the alignment of peptide sequences comprising cluster 23, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXIII and Subformula XXIIIa.



FIG. 26 shows the alignment of peptide sequences comprising cluster 24, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXIV and Subformula XXIVa.



FIG. 27 shows the alignment of peptide sequences comprising cluster 25, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXV and Subformula XXVa.



FIG. 28 shows the alignment of peptide sequences comprising cluster 26, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XVI and Subformula XVIa.



FIG. 29 shows the alignment of peptide sequences comprising cluster 27, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XVII and Subformula XVIIa.



FIG. 30 shows the alignment of peptide sequences comprising cluster 28, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XVIII and Subformula XVIIIa.



FIG. 31 shows the alignment of peptide sequences comprising cluster 29, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XIX and Subformula XIXa.



FIG. 32 shows the alignment of peptide sequences comprising cluster 30, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXX and Subformula XXXa.



FIG. 33 shows the alignment of peptide sequences comprising cluster 31, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXI and Subformula XXXIa.



FIG. 34 shows the alignment of peptide sequences comprising cluster 32, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXII and Subformula XXXIIa.



FIG. 35 shows the alignment of peptide sequences comprising cluster 33, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXIII and Subformula XXXIIIa.



FIG. 36 shows the alignment of peptide sequences comprising cluster 34, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXIV and Subformula XXXIVa.



FIG. 37 shows the alignment of peptide sequences comprising cluster 35, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXV and Subformula XXXVa.



FIG. 38 shows the alignment of peptide sequences comprising cluster 36, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXVI and Subformula XXXVIa.



FIG. 39 shows the alignment of peptide sequences comprising cluster 37, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXVII and Subformula XXXVIIa.



FIG. 40 shows the alignment of peptide sequences comprising cluster 38, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXVIII and Subformula XXXVIIIa.



FIG. 41 shows the alignment of peptide sequences comprising cluster 39, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XXXIX and Subformula XXXIXa.



FIG. 42 shows the alignment of peptide sequences comprising cluster 40, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XL and Subformula XLa.



FIG. 43 shows the alignment of peptide sequences comprising cluster 41, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XLI and Subformula XLIa.



FIG. 44 shows the alignment of peptide sequences comprising cluster 42, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XLII and Subformula XLIIa.



FIG. 45 shows the alignment of peptide sequences comprising cluster 43, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XLIII and Subformula XLIIIa.



FIG. 46 shows the alignment of peptide sequences comprising cluster 44, based on Jalview tree analysis of the representative sample of nerve-binding peptides. The corresponding ID number is indicated for each peptide, and the “majority rule consensus” residues at each position are highlighted to identify patterns, reflected in corresponding Formula XLIV and Subformula XLIVa.





DETAILED DESCRIPTION

Custom phage display libraries were generated from randomization of a parent peptide QVPWEEPYYVVKKS (SEQ ID NO:558) under conditions resulting in a mutation rate or about 50% per residue. The libraries were used to identify peptides specifically targeting nerve versus muscle. As described herein, these screens yielded a representative set of specific nerve-binding peptides set forth in SEQ ID NOS:1-557. Alignment and cluster analysis of these peptide sequences has further revealed multiple consensus sequences, corresponding to Formulas I to XLIV (and Subformulas Ia to XLIVa).


The targeting peptides described herein may bind to motor, sensory and autonomic nerves. Nerve targeting peptides can be used for nerve targeting peptide conjugates comprising a nerve targeting peptide comprising the amino acid sequence of any one of SEQ ID NOS: 1-557 and Formulas I-XLIV (including Subformulas Ia to XLIVa); and a cargo molecule, such as a drug, fluorescent moiety, or photosensitizing agent. Nerve targeting peptide conjugates of the present disclosure may be used for delivering a cargo molecule to a neuron or nerve, identifying a neuron or nerve, delivering a drug to a neuron or nerve, or delivering a photosensitizing agent to a neuron or nerve.


The invention may be more fully appreciated by reference to the following description, including the examples. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The use of headings and subheadings provided in the sections of this specification is solely for convenience of reference and does not limit the various embodiments herein, which are to be construed by reference to the specification as a whole.


Terms and Definitions

Prior to setting forth this disclosure in more detail, it may be helpful to an understanding thereof to provide definitions of certain terms to be used herein. Additional definitions are set forth throughout this disclosure.


The term “consisting essentially of” limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel characteristics of the claimed invention. It should be understood that the terms “a” and “an” as used herein refer to “one or more” of the enumerated components.


The use of the alternative (e.g., “or”) should be understood to mean either one, both, or any combination thereof of the alternatives. Thus, in embodiments of any of the Formulas and Subformulas described herein, the selection of amino acids at any position in the peptide can be any subset or combination of choices at that position. For example, in the case of Subformula (XLIVa), the embodiments not only include those in which X3 is A, L, T, or V; but also those in which X3 is A; X3 is L; X3 is T; X3 is V; X3 is A or L, X3 is A, L, or T; X3 is L, T, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X4 or X5.


As used herein, the terms “include” and “have” are used synonymously, which terms and variants thereof are intended to be construed as non-limiting. The term “comprise” means the presence of the stated features, integers, steps, or components as referred to in the claims, but does not preclude the presence or addition of one or more other features, integers, steps, components, or groups thereof.


In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. Also, any number range recited herein relating to any physical feature, such as polymer subunits, size or thickness, are to be understood to include any integer within the recited range, unless otherwise indicated. As used herein, the term “about” means ±20% of the indicated range, value, or structure, unless otherwise indicated.


In addition, it should be understood that the individual compounds, or groups of compounds, derived from the various combinations of the structures and substituents described herein, are disclosed by the present application to the same extent as if each compound or group of compounds was set forth individually. Thus, selection of particular structures or particular substituents is within the scope of the present disclosure


As used herein, the “central nervous system” or “CNS” refers to the brain and the spinal cord, as well as the retina.


As used herein, the “peripheral nervous system” or “PNS” refers to the nerves and ganglia outside the CNS. The PNS is divided into the somatic nervous system and the autonomic nervous system. The somatic nervous system controls all voluntary muscular systems within the body, and the process of voluntary reflex arcs and is composed of afferent or sensory nerves, which relay sensation from the body to the central nervous system, and efferent or motor nerves, which send out commands from the CNS to the body, stimulating muscle contraction. The autonomic nervous system supplies smooth muscle and glands, and thus influences the function of internal organs. The autonomic nervous system acts largely unconsciously and regulates bodily functions, such as the heart rate, digestion, respiratory rate, pupillary response, urination, and sexual arousal.


As used herein, a “neuron” refers to an electrically excitable cell that processes and transmits information by electrical and chemical signaling. A typical neuron possesses a cell body (often called the soma), dendrites, and an axon.


As used herein, a “nerve” refers to an enclosed, cable-like bundle of neural axons. Each nerve is a cordlike structure that contains many axons. Each axon within the nerve is an extension of an individual neuron. Within a nerve, each axon is surrounded by a layer of connective tissue called the endoneurium. The axons are bundled together into groups called fascicles, and each fascicle is wrapped in a layer of connective tissue called the perineurium. Each nerve is covered on the outside by a dense sheath of connective tissue, the epineurium.


As used herein the term “conjugate” refers to a peptide attached to at least one cargo molecule, optionally via a linker.


The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to naturally occurring amino acid polymers as well as amino acid polymers in which one or more amino acid residues is a non-naturally occurring amino acid (e.g., an amino acid analog). The terms encompass amino acid chains of any length, including full length proteins (i.e., antigens), wherein the amino acid residues are linked by covalent peptide bonds. As used herein, the term “peptide” refers to a polymer of amino acid residues typically ranging in length from 2 to about 50 residues. Where an amino acid sequence is provided herein, L-, D-, or beta amino acid versions of the sequence are also contemplated as well as retro, inversion, and retro-inversion isoforms. Peptides also include amino acid polymers in which one or more amino acid residues is an artificial chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers. In addition, the term applies to amino acids joined by a peptide linkage or by other modified linkages (e.g., where the peptide bond is replaced by an α-ester, a /3-ester, a thioamide, phosphonamide, carbamate, hydroxylate, and the like (see, e.g., Spatola, 1983, Chem. Biochem. Amino Acids and Proteins 7, 267-357), where the amide is replaced with a saturated amine (see, e.g., Skiles et al., U.S. Pat. No. 4,496,542, which is incorporated herein by reference, and Kaltenbronn et al. 1990, pp. 969-970 in Proc. 11th American Peptide Symposium, ESCOM Science Publishers, The Netherlands, and the like).


The term “amino acid” refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. An amino acid may be an L- or D-amino acid. Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.


Amino acids are referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUP AC-IUB Biochemical Nomenclature Commission, as summarized here:


















Three Letter
One Letter



Amino Acid
Symbol
Symbol









Alanine
Ala
A



Arginine
Arg
R



Asparagine
Asn
N



Aspartic Acid
Asp
D



Cysteine
Cys
C



Glutamic Acid
Glu
E



Glutamine
Gln
Q



Glycine
Gly
G



Histidine
His
H



Isoleucine
Ile
I



Leucine
Leu
L



Lysine
Lys
K



Methionine
Met
M



Phenylalanine
Phe
F



Proline
Pro
P



Serine
Ser
S



Threonine
Thr
T



Tryptophan
Trp
W



Tyrosine
Tyr
Y



Valine
Val
V










Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.


One of skill will recognize that individual substitutions, deletions or additions to a peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a “conservatively modified variant” where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to, and do not exclude, polymorphic variants, interspecies homologs, and alleles of the invention.


In some embodiments, naturally occurring amino acids in the peptides disclosed herein may be substituted with naturally occurring non-encoded amino acids and synthetic amino acids. For example, certain commonly encountered amino acids that may provide useful substitutions include, but are not limited to, β-alanine (β-Ala) and other omega-amino acids such as 3-aminopropionic acid, 2,3-diaminopropionic acid (Dpr), 4-aminobutyric acid and so forth; α-aminoisobutyric acid (Aib); ε-aminohexanoic acid (Aha); δ-aminovaleric acid (Ava); N-methylglycine or sarcosine (MeGly); ornithine (Orn); citrulline (Cit); t-butylalanine (t-BuA); t-butylglycine (t-BuG); N-methylisoleucine (MeIle); phenylglycine (Phg); cyclohexylalanine (Cha); norleucine (Nle); naphthylalanine (Nal); 4-chlorophenylalanine (Phe(4-Cl)); 2-fluorophenylalanine (Phe(2-F)); 3-fluorophenylalanine (Phe(3-F)); 4-fluorophenylalanine (Phe(4-F)); penicillamine (Pen); 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic); β-2-thienylalanine (Thi); methionine sulfoxide (MSO); homoarginine (hArg); N-acetyl lysine (AcLys); 2,4-diaminobutyric acid (Dbu); 2,3-diaminobutyric acid (Dab); p-aminophenylalanine (Phe(pNH2)); N-methyl valine (MeVal); homocysteine (hCys), homophenylalanine (hPhe) and homoserine (hSer); hydroxyproline (Hyp), homoproline (hPro), N-methylated amino acids and peptoids (N-substituted glycines).


While in most instances, the amino acids of the nerve-binding peptides disclosed herein will be substituted with L-enantiomeric amino acids, the substitutions are not limited to L-enantiomeric amino acids. Thus, in some embodiments, an L-amino acid may be replaced with an identical D-amino acid or with a D-amino acid of the same category or subcategory.


“Sequence identity,” as used herein, refers to the percentage of amino acid residues in a single sequence that are identical with the amino acid residues in another reference polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. The percentage sequence identity values can be generated using the NCBI BLAST 2.0 software as defined by Altschul et al. (1997), Nucl. Acids Res. 25:3389-3402, with the parameters set to default values.


As used herein, the terms “label” refers to a molecule that facilitates the visualization and/or detection of a targeting molecule disclosed herein. In some embodiments, the label is a fluorescent moiety.


The phrase “specifically binds” when referring to the interaction between a peptide disclosed herein and a target (e.g., purified protein, neuron or nerve tissue, neuron or nerves, cranial neuron or nerves, central neuron or nerves, myelinated or unmyelinated neuron or nerves, or connective tissue surrounding neuron or nerves), refers to the formation of a high affinity bond between the targeting molecule and the target. Further, the term means that the peptide has low affinity for non-targets.


“Selective binding,” “selectivity,” and the like refer to the preference of agent to interact with one molecule as compared to another. Preferably, interactions between a peptide disclosed herein and a target are both specific and selective. Note that in some embodiments an agent is designed to “specifically bind” and “selectively bind” two distinct, yet similar targets without binding to other undesirable targets.


The terms “individual,” “patient,” or “subject” are used interchangeably. As used herein, they mean any mammal (i.e., species of any orders, families, and genera within the taxonomic classification animalia: chordata: vertebrata: mammalia). In some embodiments, the mammal is a cow, horse, sheep, pig, cat, dog, goat, mouse, rat, rabbit, guinea pig, non-human primate, or human. In some embodiments, the mammal is a human.


The terms “administer,” “administering”, “administration,” and the like, as used herein, refer to the methods that may be used to enable delivery of agents or compositions to the desired site of biological action. These methods include, but are not limited to topical, oral, intrarectal, intravaginal, intranasal, inhalation, parenteral injection (e.g., intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, intravascular, intrathecal, intravitreal, infusion, or local). Administration techniques that are optionally employed with the agents and methods described herein, include e.g., as discussed in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa. Administration of agents may be local or systemic. In some embodiments, administration is via systemic intravenous injection into human patients.


The term “pharmaceutically acceptable” as used herein, refers to a material that does not abrogate the biological activity or properties of the agents described herein, and is relatively nontoxic (i.e., the toxicity of the material significantly outweighs the benefit of the material). In some instances, a pharmaceutically acceptable material may be administered to an individual without causing significant undesirable biological effects or significantly interacting in a deleterious manner with any of the components of the composition in which it is contained.


As used herein, “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, and having the properties of being nontoxic and non-inflammatory in a patient. Each excipient must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Examples of pharmaceutical excipients can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water or aqueous solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as excipients, particularly for injectable solutions. Suitable pharmaceutical excipients are described in “Remington's Pharmaceutical Sciences” by E. W. Martin.


The term “surgery” as used herein, refers to any methods that may be used to manipulate, change, or cause an effect by a physical intervention. These methods include, but are not limited to open surgery, endoscopic surgery, laparoscopic surgery, minimally invasive surgery, microscopic surgery, robotic surgery, any procedures that may affect any neuron or nerves such as placement of retractors during spinal surgery, microscopic procedures, cardiac neuron or nerve ablation, epidural injection, intrathecal injections, neuron or nerve blocks, implantation of devices such as neuron or nerve stimulators and implantation of pumps. In some embodiments, the subject of the surgery is a human subject or human patient.


I. Peptide

In one aspect, the present disclosure provides peptides showing selective binding for nerves versus muscle and that can be used in nerve targeting peptide conjugates of the present disclosure.


In some embodiments, a peptide comprises or consists of an amino acid sequence set forth in any one of SEQ ID NOS:1-557. In some embodiments, the peptide comprises or consists of an amino acid sequence set forth in any one of SEQ ID NOS: 1-30. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:31-36. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:37-40. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:41-240. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:241-349. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:350-449. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:450-557.


In some embodiments, a peptide comprises or consists of the amino acid sequence having at least 75% identity to any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence having at least 80% identity to any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence having at least 85% identity to any one of SEQ ID NOS:1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence having at least 90% identity to any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence having at least 92% identity to any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence having at least 95% identity to any one of SEQ ID NOS: 1-557.


In some embodiments, a peptide comprises or consists of the amino acid sequence set forth in any one of Formulas (I)-(XLIV), including Subformulas (Ia)-(XLIVa).


Formula (I)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (I): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, E, H, L, Q, R, or Y;
    • X2 is D, F, G, I, L, N, or V;
    • X3 is A, E, H, I, L, P, Q, R, S, T, or W;
    • X4 is C, F, G, K, L, Q, R, S, V, W, or Y;
    • X5 is A, D, E, G, K, L, Q, T, V, or Y;
    • X6 is A, D, E, G, K, N, Q, R, S, V, or Y;
    • X7 is A, L, M, P, S, T, or W;
    • X8 is C, E, H, L, N, P, R, S, V, or Y;
    • X9 is A, C, D, E, H, L, N, P, S, or Y;
    • X10 is A, D, E, F, G, H, I, L, N, P, S, T, V, or Y;
    • X11 is A, D, E, I, L, Q, R, T, or V;
    • X12 is D, I, K, L, N, Q, R, S, or T;
    • X13 is A, C, D, E, H, I, K, M, N, Q, R, T, or V; and
    • X14 is P, S, T, V, or Y.


In some embodiments, a peptide of Formula (I) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 25, 28, 37, 59, 60, 67, 78, 86, 99, 101, 140, 144, 149, 158, 159, 175, 182, 187, 188, 217, 233, 241, 248, 270, 282, 287, 290, 302, 317, 350, 359, 384, 407, 410, 422, 439, 448, 451, 454, 459, 460, 476, 495, 506, 513, 519, 520, 551, or 552.


In some embodiments, a peptide of Formula (I) comprises or consists of the amino acid sequence of Subformula (Ia): H-V-X3-X4-X5-X6-P-Y-Y-X10-V-X12-X13-S, wherein:

    • X3 is A, E, H, I, L, P, Q, R, S, T, or W;
    • X4 is C, F, G, K, L, Q, R, S, V, W, or Y;
    • X5 is A, D, E, G, K, L, Q, T, V, or Y;
    • X6 is A, D, E, G, K, N, Q, R, S, V, or Y;
    • X10 is A, D, E, F, G, H, I, L, N, P, S, T, V, or Y;
    • X12 is D, I, K, L, N, Q, R, S, or T; and
    • X13 is A, C, D, E, H, I, K, M, N, Q, R, T, or V.


In some embodiments, a peptide of Formula (I) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (Ia) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X6 is E; X10 is V; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (I) and Subformula (Ia) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is A, E, H, I, L, P, Q, R, S, T, or W; but also, for example, those in which X3 is A; X3 is E; X3 is H; etc.; X3 is A or E; X3 is A, E, or H; X3 is E or H; X3 is E, H, L, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X5 or X6, etc.


Formula (II)

In some embodiments, a peptide comprises or consists of an amino acid sequence of Formula (II): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, D, E, H, K, L, N, Q, R, S, or T;
    • X2 is A, F, H, I, L, T, or V;
    • X3 is A, E, F, L, P, Q, S, or T;
    • X4 is A, E, F, G, L, M, R, S, T, V, W, or Y;
    • X5 is A, E, G, H, I, P, Q, T, V, or Y;
    • X6 is A, D, E, G, H, I, K, N, Q, or R;
    • X7 is A, H, L, P, Q, R, S, or T;
    • X8 is G, H, L, R, S, V, or Y;
    • X9 is D, E, F, L, M, N, T, V, or Y;
    • X10 is A, D, F, G, H, I, L, N, R, S, V, or Y;
    • X11 is E, F, G, K, L, M, R, or V;
    • X12 is E, H, I, K, N, P, Q, R, S, T, or V,
    • X13 is D, E, F, H, I, K, N, Q, R, S, T, or V; and
    • X14 is A, F, H, P, R, S, or T.


In some embodiments, a peptide of Formula (II) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 26, 52, 68, 83, 89, 108, 114, 119, 139, 147, 174, 177, 179, 183, 195, 197, 198, 205, 209, 225, 232, 238, 279, 285, 303, 318, 321, 322, 325, 348, 363, 383, 397, 417, 462, 464, 488, 494, 502, 518, 545, or 546.


In some embodiments, a peptide of Formula (II) comprises or consists of the amino acid sequence of Subformula (IIa): X1-X2-X3-X4-E-X6-X7-Y-Y-X10-V-X12-X13-X14, wherein:

    • X1 is A, D, E, H, K, L, N, Q, R, S, or T;
    • X2 is A, F, H, I, L, T, or V;
    • X3 is A, E, F, L, P, Q, S, or T;
    • X4 is A, E, F, G, L, M, R, S, T, V, W, or Y;
    • X6 is A, D, E, G, H, I, K, N, Q, or R;
    • X7 is A, H, L, P, Q, R, S, or T;
    • X10 is A, D, F, G, H, I, L, N, R, S, V, or Y;
    • X12 is E, H, I, K, N, P, Q, R, S, T, or V,
    • X13 is D, E, F, H, I, K, N, Q, R, S, T, or V; and
    • X14 is A, F, H, P, R, S, or T.


In some embodiments, a peptide of Formula (II) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (IIa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X6 is E; X7 is P; X10 is V; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (II) and Subformula (IIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is A, E, F, L, P, Q, S, or T; but also, for example, those in which X3 is A; X3 is E; X3 is F; etc.; X3 is A or E; X3 is A, F, or P; X3 is E or L; X3 is E, F, L, or S, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X4 or X7, etc.


Formula (III):

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (III): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is H, Q, R, or V;
    • X2 is F, G, H, I, L, or V;
    • X3 is A, H, P, or T;
    • X4 is F, G, L, M, S, T, or W;
    • X5 is A, D, G, I, K, P, Q, or V;
    • X6 is D, E, G, or P;
    • X7 is L, P, Q, R, S, or T;
    • X8 is F, H, S, or Y;
    • X9 is D, F, G, H, N, S, or Y;
    • X10 is N or V;
    • X11 is A, E, G, K, L, Q, S, V, or W;
    • X12 is A, D, E, G, K, M, N, Q, R, or T;
    • X13 is E, H, K, N, Q, R, T, or V; and
    • X14 is F, H, N, P, S, T, or Y.


In some embodiments, a peptide of Formula (III) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 3, 4, 30, 80, 94, 118, 181, 214, 224, 240, 260, 286, 289, 312, 313, 357, 377, 405, or 550.


In some embodiments, a peptide of Formula (III) comprises or consists of the amino acid sequence of Subformula (IIIa): H-V-P-X4-X5-G-P-Y-X9-V-X11-K-X13-X14, wherein:

    • X4 is F, G, L, M, S, T, or W;
    • X5 is A, D, G, I, K, P, Q, or V;
    • X9 is D, F, G, H, N, S, or Y;
    • X11 is A, E, G, K, L, Q, S, V, or W;
    • X13 is E, H, K, N, Q, R, T, or V; and
    • X14 is F, H, N, P, S, T, or Y.


In some embodiments, a peptide of Formula (III) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (IIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X9 is Y; X11 is V; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (III) and Subformula (IIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is F, G, L, M, S, T, or W; but also, for example, those in which X4 is F; X4 is G; X4 is L; etc.; X4 is F or G; X4 is F, G, or L; X4 is G or L; and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X5, X9, or X10, etc.


Formula (IV)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (IV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is E, H, K, L, Q, R, S, V, or Y;
    • X2 is A, D, F, G, H, I, L, S, V, or Y;
    • X3 is A, H, K, L, N, P, Q, R, S, T, V, or W;
    • X4 is A, C, E, F, G, I, L, M, P, R, S, T, V, W, or Y;
    • X5 is A, D, E, G, H, I, K, N, Q, S, T, V, or Y;
    • X6 is A, D, E, G, H, K, N, P, Q, S, T, V, or Y;
    • X7 is A, H, L, P, Q, R, S, or T;
    • X8 is C, D, E, F, H, I, K, L, N, S, V, or Y;
    • X9 is A, D, F, H, K, L, N, S, V, W, or Y;
    • X10 is A, E, F, G, I, L, P, R, S, V, or Y;
    • X11 is A, C, E, F, G, I, L, M, N, P, Q, S, T, V, or W;
    • X12 is A, E, H, I, K, N, P, Q, R, T, V, or W;
    • X13 is D, E, H, I, K, M, N, Q, R, S, T, or Y; and
    • X14 is A, F, H, I, M, P, R, S, T, W, or Y.


In some embodiments, a peptide of Formula (IV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 22, 36, 40, 64, 65, 69, 81, 85, 102, 111, 112, 116, 121, 129, 131, 135, 136, 142, 150, 216, 223, 234, 259, 261, 262, 266, 283, 288, 292, 296, 307, 315, 319, 345, 365, 368, 373, 378, 379, 389, 393, 416, 423, 485, 503, 504, 527, 531, 538, or 539.


In some embodiments, a peptide of Formula (IV) comprises or consists of the amino acid sequence of Subformula (IVa): H-V-X3-X4-X5-X6-X7-X8-Y-V-X11-X12-K-X14, wherein:

    • X3 is A, H, K, L, N, P, Q, R, S, T, V, or W;
    • X4 is A, C, E, F, G, I, L, M, P, R, S, T, V, W, or Y;
    • X5 is A, D, E, G, H, I, K, N, Q, S, T, V, or Y;
    • X6 is A, D, E, G, H, K, N, P, Q, S, T, V, or Y;
    • X7 is A, H, L, P, Q, R, S, or T;
    • X8 is C, D, E, F, H, I, K, L, N, S, V, or Y;
    • X11 is A, C, E, F, G, I, L, M, N, P, Q, S, T, V, or W;
    • X12 is A, E, H, I, K, N, P, Q, R, T, V, or W; and
    • X14 is A, F, H, I, M, P, R, S, T, W, or Y.


In some embodiments, a peptide of Formula (IV) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (IVa) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X11 is V; X12 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (IV) and Subformula (IVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X7 is A, H, L, P, Q, R, S, or T; but also, for example, those in which X7 is A; X7 is H; X7 is L; etc.; X7 is A or H; X7 is A, H, or L; X7 is H or L; X7 is H, L, P, or S, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X7, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X4, X5, X6, etc.


Formula (V)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (V): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, F, G, H, N, Q, R, T, or Y;
    • X2 is D, F, H, I, L, M, Q, or V;
    • X3 is D, E, G, L, P, Q, R, S, or T;
    • X4 is G, L, Q, R, S, or W;
    • X5 is A, D, E, G, H, K, Q, R, or V;
    • X6 is A, E, G, Q, or V;
    • X7 is P or R;
    • X8 is S or Y;
    • X9 is G, H, N, Q, R, S, T, or Y;
    • X10 is D, F, I, P, or V;
    • X11 is A, D, F, G, I, L, Q, or V;
    • X12 is A, E, H, K, L, N, R, S, or T;
    • X13 is K, M, N, P, Q, T, or W; and
    • X14 is A, F, I, L, P, R, S, or Y.


In some embodiments, a peptide of Formula (V) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 35, 75, 76, 107, 157, 196, 207, 215, 249, 251, 252, 272, 294, 305, 310, 311, 314, 316, 320, 347, 355, 362, 370, 385, 412, 413, 456, 498, or 549.


In some embodiments, a peptide of Formula (V) comprises or consists of the amino acid sequence of Subformula (Va): H-V-X3-W-X5-E-P-Y-Y-V-X11-X12-K-X14, wherein:

    • X3 is D, E, G, L, P, Q, R, S, or T;
    • X5 is A, D, E, G, H, K, Q, R, or V;
    • X11 is A, D, F, G, I, L, Q, or V;
    • X12 is A, E, H, K, L, N, R, S, or T; and
    • X14 is A, F, I, L, P, R, S, or Y.


In some embodiments, a peptide of Formula (V) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (Va) does not comprise or consist of the amino acid sequence, wherein X3 is P; X5 is E; X11 is V; X12 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (V) and Subformula (Va) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X5 is A, D, E, G, H, K, Q, R, or V; but also, for example, those in which X5 is A; X5 is D; X5 is E; etc.; X5 is A or D; X5 is A, D, or E; X5 is D or E; X5 is A, D, E, or R, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X5, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X11 or X12, etc.


Formula (VI)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (VI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, H, L, N, Q, or Y;
    • X2 is A, D, F, G, H, I, L, R, or V;
    • X3 is A, F, L, P, S, T, or V;
    • X4 is A, G, L, S, V, W, or Y;
    • X5 is A, D, E, I, K, Q, or V;
    • X6 is A, E, L, N, P, Q, or R;
    • X7 is A, G, H, L, P, Q, R, or S;
    • X8 is A, D, F, K, N, R, S, or Y;
    • X9 is D, E, F, K, L, N, S, or Y;
    • X10 is A, E, F, G, I, L, N, P, T, or V;
    • X11 is E, K, L, M, V, or W;
    • X12 is E, H, K, L, Q, R, S, or T;
    • X13 is E, H, K, M, N, Q, or Y; and
    • X14 is F, G, I, S, or T.


In some embodiments, a peptide of Formula (VI) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 13, 33, 91, 96, 124, 167, 185, 199, 227, 235, 326, 360, 392, 409, 440, or 530.


In some embodiments, a peptide of Formula (VI) comprises or consists of the amino acid sequence of Subformula (VIa): X1-X2-X3-X4-X5-E-X7-X8-X9-X10-X11-X12-X13-S, wherein:

    • X1 is D, H, L, N, Q, or Y;
    • X2 is A, D, F, G, H, I, L, R, or V;
    • X3 is A, F, L, P, S, T, or V;
    • X4 is A, G, L, S, V, W, or Y;
    • X5 is A, D, E, I, K, Q, or V;
    • X7 is A, G, H, L, P, Q, R, or S;
    • X8 is A, D, F, K, N, R, S, or Y;
    • X9 is D, E, F, K, L, N, S, or Y;
    • X10 is A, E, F, G, I, L, N, P, T, or V;
    • X11 is E, K, L, M, V, or W;
    • X12 is E, H, K, L, Q, R, S, or T; and
    • X13 is E, H, K, M, N, Q, or Y.


In some embodiments, a peptide of Formula (VI) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (VIa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (VI) and Subformula (VIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X1 is D, H, L, N, Q, or Y; but also, for example, those in which X1 is D; X1 is H; X1 is L; etc.; X1 is D or H; X1 is D, H, or L; X1 is H or L; X1 is H, L, N, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X1, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X2 or X3, etc.


Formula (VII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (VII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, D, E, H, K, L, N, Q, R, or Y;
    • X2 is A, C, D, E, F, G, I, L, R, S, or V;
    • X3 is A, E, H, L, P, Q, R, S, T, W, or Y;
    • X4 is C, F, G, I, L, M, S, T, V, or W;
    • X5 is D, E, G, H, Q, S, or V;
    • X6 is A, D, E, G, H, Q, T, V, or Y;
    • X7 is H, L, P, Q, R, S, T, or V;
    • X8 is C, D, E, F, H, L, N, Q, S, W, or Y;
    • X9 is C, D, G, H, N, P, Q, S, W, or Y;
    • X10 is A, D, G, I, L, P, T, or V;
    • X11 is A, C, F, G, I, K, L, N, R, S, T, or V;
    • X12 is D, K, N, Q, R, T, V, or Y;
    • X13 is D, E, I, K, L, M, N, P, Q, R, T, or V; and
    • X14 is C, F, S, V, or Y.


In some embodiments, a peptide of Formula (VII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 5, 7, 10, 16, 21, 32, 53, 55, 61, 71, 84, 106, 110, 120, 123, 128, 130, 137, 151, 152, 204, 210, 237, 258, 268, 275, 281, 308, 323, 328, 330, 334, 367, 374, 375, 380, 395, 426, 433, 434, 474, 475, 486, 489, 490, 510, 525, 528, or 529.


In some embodiments, a peptide of Formula (VII) comprises or consists of the amino acid sequence of Subformula (VIIa): Q-X2-X3-W-E-X6-P-X8-Y-V-X11-K-X13-S, wherein:

    • X2 is A, C, D, E, F, G, I, L, R, S, or V;
    • X3 is A, E, H, L, P, Q, R, S, T, W, or Y;
    • X6 is A, D, E, G, H, Q, T, V, or Y;
    • X8 is C, D, E, F, H, L, N, Q, S, W, or Y;
    • X11 is A, C, F, G, I, K, L, N, R, S, T, or V; and
    • X13 is D, E, I, K, L, M, N, P, Q, R, T, or V.


In some embodiments, a peptide of Formula (VII) does not comprise or consist of an amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (VIIa) does not comprise or consist of an amino acid sequence, wherein X2 is V; X3 is P; X6 is E; X8 is Y; X11 is V; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (VII) and Subformula (VIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, C, D, E, F, G, I, L, R, S, or V; but also, for example, those in which X2 is A; X2 is C; X2 is D; etc.; X2 is A or C; X2 is A, C, or D; X2 is D or E; X2 is A, C, D, or R, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X8, or X11, etc.


Formula (VIII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (VIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, D, E, H, L, N, Q, R, S, T, Y;
    • X2 is A, D, E, F, H, I, L, N, P, R, S, T, V, Y;
    • X3 is C, I, L, P, Q, R, S, T, V;
    • X4 is A, C, E, F, G, K, L, M, N, Q, R, S, T, V, W, Y;
    • X5 is A, D, E, G, H, K, P, Q, R, S, T, V, Y;
    • X6 is A, D, E, G, H, K, L, N, P, Q, R, S, T, V, W, Y;
    • X7 is A, D, F, H, I, L, M, P, Q, S, T, W;
    • X8 is C, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, Y;
    • X9 is A, C, D, E, F, G, H, N, S, T, Y;
    • X10 is A, D, F, G, I, K, L, M, N, P, R, S, V;
    • X11 is A, E, G, H, I, M, P, Q, V, Y;
    • X12 is A, E, H, I, K, L, N, P, Q, R, T, V;
    • X13 is A, D, E, I, K, M, N, P, R, S, T, V, Y; and
    • X14 is A, C, F, G, H, L, N, P, S, T, Y.


In some embodiments, a peptide of Formula (VIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 1, 6, 12, 14, 19, 27, 38, 43, 44, 45, 50, 66, 90, 103, 104, 109, 132, 145, 160, 164, 166, 168, 170, 178, 191, 194, 213, 219, 220, 221, 247, 255, 257, 265, 267, 284, 298, 299, 336, 339, 340, 343, 349, 353, 372, 382, 406, 411, 415, 428, 431, 435, 445, 447, 453, 468, 471, 472, 491, 509, 511, 512, 522, 547 and 555.


In some embodiments, a peptide of Formula (VIII) comprises or consists of the amino acid sequence of Subformula (VIIIa): Q-X2-P-X4-X5-X6-P-X8-Y-X10-V-X12-X13-X14, wherein:

    • X2 is A, D, E, F, H, I, L, N, P, R, S, T, V, Y;
    • X4 is A, C, E, F, G, K, L, M, N, Q, R, S, T, V, W, Y;
    • X5 is A, D, E, G, H, K, P, Q, R, S, T, V, Y;
    • X6 is A, D, E, G, H, K, L, N, P, Q, R, S, T, V, W, Y;
    • X8 is C, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, Y;
    • X10 is A, D, F, G, I, K, L, M, N, P, R, S, V;
    • X12 is A, E, H, I, K, L, N, P, Q, R, T, V;
    • X13 is A, D, E, I, K, M, N, P, R, S, T, V, Y; and
    • X14 is A, C, F, G, H, L, N, P, S, T, Y.


In some embodiments, a peptide of Formula (VIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (VIIIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X5 is E; X6 is E; X8 is Y; X10 is V; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (VIII) and Subformula (VIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, D, E, F, H, I, L, N, P, R, S, T, V, Y; but also, for example, those in which X2 is A; X2 is D; X2 is E; etc.; X2 is A or D; X2 is A, D, or E; X2 is D or E; X2 is D, E, F, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X12, or X13, etc.


Formula (IX)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (IX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, H, N, Q, R, S, W, or Y;
    • X2 is D, F, H, I, L, M, N, P, R, S, V, or Y;
    • X3 is A, E, H, L, P, Q, R, S, T, or Y;
    • X4 is A, G, H, I, L, N, P, R, S, or W;
    • X5 is A, E, G, I, K, Q, S, or V;
    • X6 is A, D, E, G, K, L, P, Q, R, T, or V;
    • X7 is A, G, K, L, P, Q, R, S, or T;
    • X8 is E, I, K, L, S, V, or Y;
    • X9 is A, D, F, G, H, I, K, N, Q, S, T, W, or Y;
    • X10 is A, D, F, H, I, L, N, T, or V;
    • X11 is A, D, E, F, G, L, M, V, or W;
    • X12 is E, K, N, Q, R, T, or V;
    • X13 is E, H, K, L, N, Q, R, S, T, or Y; and
    • X14 is E, P, R, S, T, or Y.


In some embodiments, a peptide of Formula (IX) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 8, 31, 79, 93, 127, 153, 155, 156, 169, 186, 189, 202, 212, 218, 226, 304, 329, 331, 335, 338, 352, 361, 366, 371, 376, 381, 390, 401, 408, 418, 424, 430, 436, 438, 443, 457, 493, 515, 521, 544.


In some embodiments, a peptide of Formula (IX) comprises or consists of the amino acid sequence of Subformula (IXa): Q-X2-P-W-E-X6-P-Y-X9-X10-X11-X12-X13-S, wherein:

    • X2 is D, F, H, I, L, M, N, P, R, S, V, or Y;
    • X6 is A, D, E, G, K, L, P, Q, R, T, or V;
    • X9 is A, D, F, G, H, I, K, N, Q, S, T, W, or Y;
    • X10 is A, D, F, H, I, L, N, T, or V;
    • X11 is A, D, E, F, G, L, M, V, or W;
    • X12 is E, K, N, Q, R, T, or V; and
    • X13 is E, H, K, L, N, Q, R, S, T, or Y.


In some embodiments, a peptide of Formula (IX) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E, X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (IXa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X6 is E; X7 is P; X9 is Y; X10 is V; Xu is V; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (IX) and Subformula (IXa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X12 is E, K, N, Q, R, T, or V; but also, for example, those in which X12 is E; X12 is K; X12 is N; etc.; X12 is E or K; X12 is E, K, or N; X12 is K or N; X12 is K, N, Q, or T, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X12, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X6, or X13, etc.


Formula (X)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (X): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, D, E, G, H, K, N, Q, R, S, T, or Y;
    • X2 is A, D, F, H, I, L, M, Q, S, T, V, or Y;
    • X3 is A, E, G, K, L, N, P, Q, R, S, T, V, or Y;
    • X4 is C, F, G, H, L, M, R, S, T, W, or Y;
    • X5 is A, D, E, G, K, L, N, Q, T, or V;
    • X6 is A, D, E, G, H, K, L, N, Q, S, or V,
    • X7 is A, L, P, R, S, T, or Y;
    • X8 is A, C, D, F, G, N, R, S, T, or Y;
    • X9 is A, D, F, H, L, N, Q, R, S, T, or Y;
    • X10 is A, D, E, F, G, H, I, L, P, R, S, T, or V;
    • X11 is A, D, E, F, G, H, I, L, M, P, Q, R, S, or V;
    • X12 is A, D, E, G, H, K, N, Q, S, T, or V;
    • X13 is D, E, F, H, I, K, L, M, N, Q, R, S, T, V, or Y; and
    • X14 is A, F, H, I, K, L, N, P, R, S, T, V, or Y.


In some embodiments, a peptide of Formula (X) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 2, 17, 20, 23, 39, 41, 42, 56, 58, 63, 70, 97, 113, 125, 133, 134, 141, 148, 163, 165, 176, 180, 193, 203, 222, 228, 229, 243, 245, 254, 277, 295, 297, 309, 324, 342, 344, 346, 358, 387, 391, 396, 421, 425, 429, 432, 458, 461, 465, 467, 473, 477, 487, 514, 532, 537, 540, and 556.


In some embodiments, a peptide of Formula (X) comprises or consists of the amino acid sequence of Subformula (Xa): Q-X2-X3-X4-X5-E-P-Y-Y-X10-X11-K-X13-X14, wherein:

    • X2 is A, D, F, H, I, L, M, Q, S, T, V, or Y;
    • X3 is A, E, G, K, L, N, P, Q, R, S, T, V, or Y;
    • X4 is C, F, G, H, L, M, R, S, T, W, or Y;
    • X5 is A, D, E, G, K, L, N, Q, T, or V;
    • X10 is A, D, E, F, G, H, I, L, P, R, S, T, or V;
    • X11 is A, D, E, F, G, H, I, L, M, P, Q, R, S, or V;
    • X13 is D, E, F, H, I, K, L, M, N, Q, R, S, T, V, or Y; and
    • X14 is A, F, H, I, K, L, N, P, R, S, T, V, or Y.


In some embodiments, a peptide of Formula (X) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (Xa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X4 is W; X5 is E; X10 is V; X11 is V; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (X) and Subformula (Xa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X5 is A, D, E, G, K, L, N, Q, T, or V; but also, for example, those in which X5 is A; X5 is D; X5 is E; etc.; X5 is A or D; X5 is A, D, or E; X5 is D or E; X5 is D, E, G, or K and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X5, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X2, or X3, etc.


Formula (XI)

In some embodiments, a peptide comprises or consists of an amino acid sequence of Formula (XI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, H, L, Q, or R;
    • X2 F, G, H, I, L, N, Q, S, T, V, or Y;
    • X3 is A, G, H, L, P, Q, R, S, T, or Y;
    • X4 is A, C, F, G, L, P, Q, R, S, W, or Y;
    • X5 is A, D, E, G, H, K, N, P, Q, S, T, V, or Y;
    • X6 is A, D, E, G, H, K, L, N, Q, T, or V;
    • X7 is A, E, H, K, L, P, Q, R, S, or T;
    • X8 is C, D, E, F, G, H, K, L, N, P, Q, S, T, V, or Y;
    • X9 is A, D, F, G, H, I, L, N, Q, S, T, V, W, or Y;
    • X10 is A, D, F, G, H, I, L, P, R, S, T, V, or Y;
    • X11 is A, D, E, F, G, H, L, M, Q, S, or V;
    • X12 is A, E, H, I, K, L, N, P, Q, R, S, or T;
    • X13 is A, D, H, I, K, L, M, N, P, Q, S, T, V, or Y; and
    • X14 is A, F, H, L, P, S, T, V, or Y.


In some embodiments, a peptide of Formula (XI) comprises or consists of an amino acid sequence of any one of SEQ ID NOS: 11, 29, 46, 48, 51, 62, 77, 87, 88, 95, 98, 100, 162, 173, 201, 236, 253, 256, 263, 273, 278, 300, 327, 341, 351, 364, 386, 388, 399, 400, 403, 404, 414, 419, 420, 437, 441, 444, 449, 450, 466, 469, 478, 481, 483, 484, 496, 500, 516, 524, 533, 535, 536, 542, 543, 548, and 554.


In some embodiments, a peptide of Formula (XI) comprises or consists of an amino acid sequence of Subformula (XIa): Q-X2-P-X4-X5-X6-X7-X8-Y-X10-X11-K-X13-S, wherein:

    • X2 is F, G, H, I, L, N, Q, S, T, V, or Y;
    • X4 is A, C, F, G, L, P, Q, R, S, W, or Y;
    • X5 is A, D, E, G, H, K, N, P, Q, S, T, V, or Y;
    • X6 is A, D, E, G, H, K, L, N, Q, T, or V;
    • X7 is A, E, H, K, L, P, Q, R, S, or T;
    • X8 is C, D, E, F, G, H, K, L, N, P, Q, S, T, V, or Y;
    • X10 is A, D, F, G, H, I, L, P, R, S, T, V, or Y;
    • X11 is A, D, E, F, G, H, L, M, Q, S, or V; and
    • X13 is A, D, H, I, K, L, M, N, P, Q, S, T, V, or Y.


In some embodiments, a peptide of Formula (XI) does not comprise or consist of an amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XIa) does not comprise or consist of an amino acid sequence, wherein X2 is V; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X10 is V; X11 is V; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XI) and Subformula (XIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X7 is A, E, H, K, L, P, Q, R, S, or T; but also, for example, those in which X7 is A; X7 is H; X7 is L; etc.; X7 is E or H; X7 is E, H, or L; X7 is H or L; X7 is H, L, P, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X7, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X10, or X11, etc.


Formula (XII)

In some embodiments, a peptide comprises or consists of an amino acid sequence of Formula (XII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is E, H, K, L, N, Q, or R;
    • X2 is A, C, D, F, I, K, L, M, N, S, V, or Y;
    • X3 is A, L, P, R, S, or T;
    • X4 is A, E, F, G, L, M, R, S, T, W, or Y;
    • X5 is A, D, E, F, G, H, K, N, Q, R, T, V, or Y;
    • X6 is A, D, E, G, Q, R, S, T, V, or Y;
    • X7 is A, H, K, L, P, Q, R, S, or T;
    • X8 is A, C, D, F, H, N, R, S, T, or Y;
    • X9 is D, E, F, H, L, N, R, S, T, V, or Y;
    • X10 is A, D, F, G, H, I, L, N, R, S, T, or V;
    • X11 is A, D, F, G, L, M, P, Q, R, T, V, or W;
    • X12 is A, D, E, H, K, L, M, N, P, Q, R, S, T, or Y;
    • X13 is E, H, I, K, L, M, N, P, Q, R, S, T, or V; and
    • X14 is A, F, P, or S.


In some embodiments, a peptide of Formula (XII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 9, 34, 47, 57, 72, 73, 82, 92, 105, 115, 117, 122, 138, 143, 146, 161, 171, 172, 190, 200, 208, 211, 244, 246, 274, 276, 280, 293, 332, 333, 337, 354, 356, 369, 394, 398, 402, 442, 446, 452, 455, 463, 470, 479, 480, 482, 497, 499, 501, 507, 508, 517, 534, 541, 553, and 557.


In some embodiments, a peptide of Formula (XII) comprises or consists of the amino acid sequence of Subformula (XIIa): Q-X2-P-X4-X5-E-P-Y-X9-X10-X11-X12-X13-S, wherein:

    • X2 is A, C, D, F, I, K, L, M, N, S, V, or Y;
    • X4 is A, E, F, G, L, M, R, S, T, W, or Y;
    • X5 is A, D, E, F, G, H, K, N, Q, R, T, V, or Y;
    • X9 is D, E, F, H, L, N, R, S, T, V, or Y;
    • X10 is A, D, F, G, H, I, L, N, R, S, T, or V;
    • X11 is A, D, F, G, L, M, P, Q, R, T, V, or W;
    • X12 is A, D, E, H, K, L, M, N, P, Q, R, S, T, or Y; and
    • X13 is E, H, I, K, L, M, N, P, Q, R, S, T, or V.


In some embodiments, a peptide of Formula (XII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XIIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X5 is E; X9 is Y; X10 is V; X11 is V; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XII) and Subformula (XIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, C, D, F, I, K, L, M, N, S, V, or Y; but also, for example, those in which X2 is A; X2 is C; X2 is D; etc.; X2 is A or C; X2 is A, C, or D; X2 is C or D; X2 is C, D, F, or L, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X4, or X5, etc.


Formula (XIII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XIII): X1-X2-X3-X4-X5-E-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is E, F, H, or Q;
    • X2 is A, F, I, M, S, T, or V;
    • X3 is A, G, H, P, Q, R, or T;
    • X4 is F, G, L, M, N, R, S, W, or Y;
    • X5 is A, G, K, N, P, Q, or R;
    • X7 is H, P, R, or T;
    • X8 is D, F, H, K, L, N, or Y;
    • X9 is F, G, H, K, Q, T, V, or Y;
    • X10 is F, G, L, P, S, V, or Y;
    • X11 is L, M, Q, or V;
    • X12 is E, G, I, K, L, N, P, S, or Y;
    • X13 is E, H, I, N, Q, S, or T; and
    • X14 is I, P, S, or Y.


In some embodiments, a peptide of Formula (XIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 15, 54, 74, 154, 184, 192, 206, 230, 242, 269, 271, 306, 427, 492, or 505.


In some embodiments, a peptide of Formula (XIII) comprises or consists of the amino acid sequence of Subformula (XIIIa): H-X2-A-X4-X5-E-P-X8-X9-X10-V-X12-X13-S, wherein:

    • X2 is A, F, I, M, S, T, or V;
    • X4 is F, G, L, M, N, R, S, W, or Y;
    • X5 is A, G, K, N, P, Q, or R;
    • X8 is D, F, H, K, L, N, or Y;
    • X9 is F, G, H, K, Q, T, V, or Y;
    • X10 is F, G, L, P, S, V, or Y;
    • X12 is E, G, I, K, L, N, P, S, or Y; and
    • X13 is E, H, I, N, Q, S, or T.


In some embodiments, a peptide of Formula (XIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XIIIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X8 is Y; X9 is Y; X10 is V; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XIII) and Subformula (XIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, F, I, M, S, T, or V; but also, for example, those in which X2 is A; X2 is F; X2 is I; etc.; X2 is A or F; X2 is A, F, or I; X2 is F or I; X2 is F, I, M, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X8 or X9, etc.


Formula (XIV)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XIV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is H, L, N, Q, or R;
    • X2 is F, L, R, S, or V;
    • X3 is P, S, or T;
    • X4 is A, L, S, or T;
    • X5 is A, D, E, H, Q, R, V, or Y;
    • X6 is D, E, or K;
    • X7 is A, P, Q, or W;
    • X8 is H, R, W, or Y;
    • X9 is H, K, L, S, or Y;
    • X10 is D, E, F, L, T, or V;
    • X11 is A, D, or L;
    • X12 is D, E, G, H, K, N, Q, or R;
    • X13 is G, K, N, Q, or T; and
    • X14 is P or S.


In some embodiments, a peptide of Formula (XIV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 18, 24, 49, 126, 231, 239, 250, 264, 291, 301, 523, and 526. In some embodiments, a peptide of Formula (XIV) comprises or consists of the amino acid sequence of Subformula (XIVa): H-V-X3-S-X5-E-P-Y-Y-X10-L-X12-X13-S, wherein:

    • X3 is P, S, or T;
    • X5 is A, D, E, H, Q, R, V, or Y;
    • X10 is D, E, F, L, T, or V;
    • X12 is D, E, G, H, K, N, Q, or R; and
    • X13 is G, K, N, Q, or T.


In some embodiments, a peptide of Formula (XIV) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XIVa) does not comprise or consist of the amino acid sequence, wherein X3 is P; X5 is E; X10 is V; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XIV) and Subformula (XIVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is P, S, or T; but also, for example, those in which X3 is P; X3 is S; X3 is T; etc.; X3 is Por S; X3 is P, S, or T; X3 is S or T; and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X12, or X13, etc.


Formula (XV)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XV): X1-X2-X3-X4-X5-X6-X7-Y-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is E, K, Q, or T;
    • X2 is D, F, L, P, or V;
    • X3 is C, L, P, T, or V;
    • X4 is A, L, or T;
    • X5 is D, E, H, or Q;
    • X6 is A, D, K, or N;
    • X7 is A, P, or S;
    • X9 is C, H, M, or Y;
    • X10 is F, L, or V;
    • X11 is E, G, H, R, or V;
    • X12 is K or T;
    • X13 is I or Q; and
    • X14 is A, F, or T.


In some embodiments, a peptide of Formula (XV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 150, 238, 321, 397, and 415.


In some embodiments, a peptide of Formula (XV) comprises or consists of the amino acid sequence of Subformula (XVa): X1-X2-X3-X4-X5-X6-P-Y-X9-X10-X11-T-Q-T. wherein:

    • X1 is E, K, Q, or T;
    • X2 is D, F, L, P, or V;
    • X3 is C, L, P, T, or V;
    • X4 is A, L, or T;
    • X5 is D, E, H, or Q;
    • X6 is A, D, K, or N;
    • X9 is C, H, M, or Y;
    • X10 is F, L, or V; and
    • X11 is E, G, H, R, or V.


In some embodiments, a peptide of Formula (XV) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X5 is E; X7 is P; X9 is Y; X10 is V; Xu is V; and X12 is K.


In some embodiments, a peptide of Subformula (XVa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X5 is E; X9 is Y; X10 is V; and X11 is V.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XV) and Subformula (XVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X1 is E, K, Q, or T; but also, for example, those in which X1 is E; X1 is K; X1 is Q; etc.; X1 is E or K; X1 is E, K, or Q; X1 is E or K; X1 is E, K, or T; and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X1, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X2 or X3, etc.


Formula (XVI)

In some embodiments, a peptide comprises or consists of an amino acid sequence of Formula (XVI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, H, K, or Q;
    • X2 is F, I, or V;
    • X3 is A, P, R, S, or T;
    • X4 is G, L, M, Q, S, T, or W;
    • X5 is D, G, Q, T, or V;
    • X6 is D, E, H, Q, or S;
    • X7 is A, P, R, or T;
    • X8 is E, G, T, V, or Y;
    • X9 is A, Q, S, or Y;
    • X10 is A, D, E, G, I, L, R, T, V, or Y;
    • X11 is A, E, F, G, or I;
    • X12 is K, Q, or T;
    • X13 is E, I, K, M, N, or S; and
    • X14 is F, L, S, or T.


In some embodiments, a peptide of Formula (XVI) comprises or consists of an amino acid sequence of any one of SEQ ID NOS: 2, 48, 63, 70, 100, 141, 193, 228, 292, 403, 425, 473, and 477.


In some embodiments, the peptide comprises or consists of the amino acid sequence of Subformula (XVIa): Q-F-P-X4-X5-E-P-Y-Y-X10-X11-K-K-T, wherein:

    • X4 is G, L, M, Q, S, T, or W;
    • X5 is D, G, Q, T, or V;
    • X10 is A, D, E, G, I, L, R, T, V, or Y; and
    • X11 is A, E, F, G, or I.


In some embodiments, a peptide of Formula (XVI) does not comprise or consist of an amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XVIa) does not comprise or consist of an amino acid sequence, wherein X4 is W and X10 is V.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XVI) and Subformula (XVIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X5 is D, G, Q, T, or V; but also, for example, those in which X5 is D; X5 is G; X5 is Q; etc.; X5 is D or G; X5 is D, G, or Q; X5 is Q or T; X5 is D, Q, T, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X5, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X10 or X11, etc.


Formula (XVII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XVII): Q-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X2 is H, S, or T;
    • X3 is P, Q, or T;
    • X4 is F, M, P, or W;
    • X5 is D, E, or Q;
    • X6 is E or P;
    • X7 is P, R, or T;
    • X8 is A, K, or Y;
    • X9 is H or Y;
    • X10 is G, R, or V;
    • X11 is L or V;
    • X12 is I, L, S, or T;
    • X13 is H, K, M, or Q; and
    • X14 is A or S.


In some embodiments, a peptide of Formula (XVII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 74, 265, 266, 296, and 482.


In some embodiments, a peptide of Formula (XVII) comprises or consists of the amino acid sequence of Subformula (XVIIa): Q-S-P-X4-D-E-X7-Y-Y-G-V-X12-X13-A, wherein:

    • X4 is F, M, P, or W;
    • X7 is P, R, or T;
    • X12 is I, L, S, or T; and
    • X13 is H, K, M, or Q.


In some embodiments, a peptide of Formula (XVII) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XVIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X7 is P; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XVII) and Subformula (XVIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is F, M, P, or W; but also, for example, those in which X4 is F; X4 is M; X4 is W; etc.; X4 is F or M; X4 is F, M, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X12 or X13, etc.


Formula (XVIII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XVIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is K, Q, or R;
    • X2 is F or L;
    • X3 is G, P, or T;
    • X4 is E, L, Q, S, or W;
    • X5 is A, D, E, F, P, Q, V, or Y;
    • X6 is A, D, E, K, N, Q, T, or Y;
    • X7 is H, L, P, Q, R, or T;
    • X8 is F, N, or Y;
    • X9 is H, T, or Y;
    • X10 is F, H, I, L, N, T, or Y;
    • X11 is D, G, L, M, Q, or V;
    • X12 is D, E, K, R, or T;
    • X13 is A, E, K, M, N, P, Q, R, or T; and
    • X14 is A, S, or P.


In some embodiments, the peptide of Formula (XVIII) comprises or consists of the amino acid of any one of SEQ ID NOS: 50, 88, 117, 143, 201, 211, 300, 303, 337, 420, 455, 463, 499, and 536.


In some embodiments, a peptide of Formula (XVIII) comprises or consists of the amino acid sequence of Subformula (XVIIIa): Q-L-P-L-X5-X6-X7-N-Y-X10-V-T-X13-S, wherein:

    • X5 is A, D, E, F, P, Q, V, or Y;
    • X6 is A, D, E, K, N, Q, T, or Y;
    • X7 is H, L, P, Q, R, or T;
    • X10 is F, H, I, L, N, T, or Y; and
    • X13 is A, E, K, M, N, P, Q, R, or T.


In some embodiments, a peptide of Formula (XVIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XVIIIa) does not comprise or consist of the amino acid sequence, wherein X5 is E; X6 is E; X7 is P; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XVIII) and Subformula (XVIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X5 is A, D, E, F, P, Q, V, or Y; but also, for example, those in which X5 is A; X5 is D; X5 is E; etc.; X5 is A or D; X5 is A, D, or E; X5 is D or E; X5 is D, E, P, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X5, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X10 or X13, etc.


Formula (XIX)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XIX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is Q or T;
    • X2 is H, I, L, T, or V;
    • X3 is P or T;
    • X4 is A, C, F, L, R, S, T, or Y;
    • X5 is A, E, G, H, or Q;
    • X6 is A, E, K, Q, T, V, or Y;
    • X7 is L, M, P, Q, or S;
    • X8 is C, F, L, N, W, or Y;
    • X9 is C or Y;
    • X10 is A, L, P, S, or V;
    • X11 is A, G, or V;
    • X12 is K, N, Q, R, or V;
    • X13 is A, E, K, T, V, or Y; and
    • X14 is F, G, N, P, S, T, or Y.


In some embodiments, a peptide of Formula (XIX) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 1, 6, 12, 45, 103, 220, 267, 298, 299, 336, 343, 428, 491, and 522.


In some embodiments, a peptide of Formula (XIX) comprises or consists of the amino acid sequence of Subformula (XIXa): Q-X2-P-X4-E-X6-P-X8-Y-X10-V-X12-K-X14, wherein:

    • X2 is H, I, L, T, or V;
    • X4 is A, C, F, L, R, S, T, or Y;
    • X6 is A, E, K, Q, T, V, or Y;
    • X8 is C, F, L, N, W, or Y;
    • X10 is A, L, P, S, or V;
    • X12 is K, N, Q, R, or V; and
    • X14 is F, G, N, P, S, T, or Y.


In some embodiments, a peptide of Formula (XIX) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XIXa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X6 is E; X8 is Y; X10 is V; X12 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XIX) and Subformula (XIXa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is H, I, L, T, or V; but also, for example, those in which X2 is H; X2 is I; X2 is L; etc.; X2 is H or I; X2 is H, I, or L; X2 is I or L; X2 is I, L, T, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X12 or X14, etc.


Formula (XX)

In some embodiments, the peptide comprises or consists of an amino acid sequence of Formula (XX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is E, H, N, Q, or S;
    • X2 is H, I, L, or V;
    • X3 is A, P, or T;
    • X4 is G, L, M, or W;
    • X5 is D, E, G, or V;
    • X6 is A, D, E, G, H, L, or V;
    • X7 is G, K, P, Q, R, or T;
    • X8 is E, L, S, or Y;
    • X9 is H, K, N, Q, R, S, V, or Y;
    • X10 is A, F, I, L, or P;
    • X11 is F, G, I, L, M, P, R, or V;
    • X12 is H, K, N, or V;
    • X13 is H, K, L, M, R, S, or T; and
    • X14 is F, P, R, S, or Y.


In some embodiments, a peptide of Formula (XX) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 31, 58, 92, 105, 125, 169, 176, 245, 276, 293, 338, 381, 443, 452, 497, and 501.


In some embodiments, a peptide of Formula (XX) comprises or consists of the amino acid sequence of Subformula (XXa): Q-L-P-G-E-X6-X7-Y-X9-L-X11-K-K-S, wherein:

    • X6 is A, D, E, G, H, L, or V;
    • X7 is G, K, P, Q, R, or T;
    • X9 is H, K, N, Q, R, S, V, or Y; and
    • X11 is F, G, I, L, M, P, R, or V.


In some embodiments, a peptide of Formula (XX) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXa) does not comprise or consist of an amino acid sequence, wherein X6 is E; X7 is P; X9 is Y; and X11 is V.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XX) and Subformula (XXa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X1 is E, H, N, Q, or S; but also, for example, those in which X1 is E; X1 is H; X1 is Q; etc.; X1 is E or H; X1 is E, H, or N; X1 is N or Q; X1 is N, Q, or S, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X1, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X6 or X7, etc.


Formula (XXI)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXI): X1-X2-X3-X4-X5-E-X7-X8-X9-X10-X11-K-X13-X14, wherein:

    • X1 is D, Q, or R;
    • X2 is A, F, H, I, L, V, or Y;
    • X3 is A, E, L, P, or Q;
    • X4 is A, L, R, S, or V;
    • X5 is A, E, G, H, K, Q, or R;
    • X7 is K, L, P, R, or T;
    • X8 is D, N, R, or Y;
    • X9 is E, F, H, N, R, S, or T;
    • X10 is A, D, F, I, L, R, or V;
    • X11 is A, F, L, M, P, Q, V, or W;
    • X13 is E, H, K, M, or R; and
    • X14 is P, F, or S.


In some embodiments, a peptide of Formula (XXI) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 9, 57, 138, 146, 163, 203, 227, 244, 274, 354, 442, 446, 487, and 532.


In some embodiments, a peptide of Formula (XXI) comprises or consists of the amino acid sequence of Subformula (XXIa): Q-X2-P-S-X5-E-P-Y-X9-X10-X11-K-X13-S, wherein:

    • X2 is A, F, H, I, L, V, or Y;
    • X5 is A, E, G, H, K, Q, or R;
    • X9 is E, F, H, N, R, S, or T;
    • X10 is A, D, F, I, L, R, or V;
    • X11 is A, F, L, M, P, Q, V, or W; and
    • X13 is E, H, K, M, or R.


In some embodiments, a peptide of Formula (XXI) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X5 is E; X7 is P; X8 is Y; X10 is V; X11 is V; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXI) does not comprise or consist of the amino acid sequence, wherein X2 is V; X5 is E; X10 is V; X is V; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXI) and Subformula (XXIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, F, H, I, L, V, or Y; but also, for example, those in which X2 is A; X2 is F; X2 is H; etc.; X2 is A or F; X2 is A, F, or H; X2 is F or H; X2 is F, H, I, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X10 or X11, etc.


Formula (XXII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XXII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, H, L, or Q;
    • X2 is D, F, I, L, N, S, T, or Y;
    • X3 is P, Q, S, or T;
    • X4 is A, C, L, S, or W;
    • X5 is A, D, E, H, K, Q, S, T, or V;
    • X6 is A, D, E, G, H, K, Q, R, S, W, or Y;
    • X7 is D, P, or S;
    • X8 is C, F, G, H, L, N, Q, S, T, W, or Y;
    • X9 is A, C, D, F, N, S, or Y;
    • X10 is A, D, G, L, or V;
    • X11 is A, G, L, Q, S, T, V, W, or Y;
    • X12 is A, D, K, P, Q, S, T, or Y;
    • X13 is I, K, M, N, P, R, or T; and
    • X14 is A, C, F, P, S, or Y.


In some embodiments, a peptide of Formula (XXII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 7, 16, 19, 27, 47, 66, 72, 166, 194, 236, 246, 278, 332, 340, 394, 453, 475, and 479.


In some embodiments, a peptide of Formula (XXII) comprises or consists of the amino acid sequence of Subformula (XXIIa): Q-X2-P-X4-X5-X6-P-X8-S-X10-V-X12-X13-X14, wherein:

    • X2 is D, F, I, L, N, S, T, or Y;
    • X4 is A, C, L, S, or W;
    • X5 is A, D, E, H, K, Q, S, T, or V;
    • X6 is A, D, E, G, H, K, Q, R, S, W, or Y;
    • X8 is C, F, G, H, L, N, Q, S, T, W, or Y;
    • X10 is A, D, G, L, or V;
    • X12 is A, D, K, P, Q, S, T, or Y;
    • X13 is I, K, M, N, P, R, or T; and
    • X14 is A, C, F, P, S, or Y.


In some embodiments, a peptide of Formula (XXII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X5 is E; X6 is E; X8 is Y; X10 is V; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXII) and Subformula (XXIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is A, C, L, S, or W; but also, for example, those in which X4 is C; X4 is L; X4 is S; etc.; X4 is C or L; X4 is C, L, or S; X4 is L or S; X4 is C, L, S, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X6 or X8, etc.


Formula (XXIII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, or Q;
    • X2 is L, M, N, P, R, or V;
    • X3 is P, Q, R, or T;
    • X4 is R, S, or W;
    • X5 is E, G, H, Q, T, or Y;
    • X6 is A, E, K, N, Q, or V;
    • X7 is A, P, R, or S;
    • X8 is D, F, H, K, N, or Y;
    • X9 is D, F, H, N, S, T, W, or Y;
    • X10 is G, I, R, or V;
    • X11 is D, G, M, or V;
    • X12 is E, K, N, R, or V;
    • X13 is I, K, N, Q, R, S, T, V, or Y; and
    • X14 is G, H, P, S, T, or Y.


In some embodiments, a peptide of Formula (XXIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 46, 87, 93, 98, 202, 218, 255, 339, 351, 352, 361, 376, 430, 436, 496, and 511.


In some embodiments, a peptide of Formula (XXIII) comprises or consists of the amino acid sequence of Subformula (XXIIIa): Q-L-P-W-X5-X6-P-Y-X9-V-V-X12-X13-S, wherein:

    • X5 is E, G, H, Q, T, or Y;
    • X6 is A, E, K, N, Q, or V;
    • X9 is D, F, H, N, S, T, W, or Y;
    • X12 is E, K, N, R, or V; and
    • X13 is I, K, N, Q, R, S, T, V, or Y.


In some embodiments, a peptide of Formula (XXIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXIIIa) does not comprise or consist of the amino acid sequence, wherein X5 is E; X6 is E; X9 is Y; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXIII) and Subformula (XXIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X5 is E, G, H, Q, T, or Y; but also, for example, those in which X5 is E; X5 is G; X5 is H; etc.; X5 is E or G; X5 is E, G, or H; X5 is G or H; X5 is G, H, Q, or T, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X5, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X12 or X13, etc.


Formula (XXIV)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXIV): Q-X2-P-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-S wherein:

    • X2 is D, I, K, L, or M;
    • X4 is F, G, L, S, or T;
    • X5 is E, G, H, N, Q, V, or Y;
    • X6 is A, D, E, G, or S;
    • X7 is L, P, or S;
    • X8 is N, T, or Y;
    • X9 is D, N, or Y;
    • X10 is A, G, I, S, or V;
    • X11 is D, G, L, Q, or T;
    • X12 is K, L, M, N, Q, R, S, or T; and
    • X13 is E, K, M, N, P, S, or V.


In some embodiments, a peptide of Formula (XXIV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 115, 172, 208, 333, 356, 398, 470, 480, 508, 517, 553, and 557.


In some embodiments, a peptide of Formula (XXIV) comprises or consists of the amino acid sequence of Subformula (XXIVa): Q-X2-P-X4-X5-X6-P-Y-Y-V-X11-X12-X13-S, wherein:

    • X2 is D, I, K, L, or M;
    • X4 is F, G, L, S, or T;
    • X5 is E, G, H, N, Q, V, or Y;
    • X6 is A, D, E, G, or S;
    • X11 is D, G, L, Q, or T;
    • X12 is K, L, M, N, Q, R, S, or T; and
    • X13 is E, K, M, N, P, S, or V.


In some embodiments, a peptide of Formula (XXIV) does not comprise or consist of the amino acid sequence, wherein X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X12 is K; and X13 is K.


In some embodiments, a peptide of Subformula (XXIVa) does not comprise or consist of the amino acid sequence, wherein X5 is E; X6 is E; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXIV) and Subformula (XXIVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is D, I, K, L, or M; but also, for example, those in which X2 is D; X2 is I; X2 is K; etc.; X2 is D or I; X2 is D, I, or K; X2 is I or K; X2 is D, I, K, or M, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X11 or X12, etc.


Formula (XXV)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXV): Q-X2-P-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X2 is F, I, L, N, S, or V;
    • X4 is E, G, L, R, W, or Y;
    • X5 is A, D, G, H, K, N, Q, S, T, V, or Y;
    • X6 is A, D, E, H, N, Q, S, or T;
    • X7 is A, K, L, P, or R;
    • X8 is N, Q, S, T, or Y;
    • X9 is E, F, H, N, V, or Y;
    • X10 is S or V;
    • X11 is F, H, L, M, S, or V;
    • X12 is E, H, I, K, or P;
    • X13 is K, N, Q, S, or Y; and
    • X14 is F, S, V, or Y.


In some embodiments, a peptide of Formula (XXV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 11, 29, 77, 82, 263, 280, 369, 402, 419, 449, 466, 478, 507, 534, 542, and 554.


In some embodiments, a peptide of Formula (XXV) comprises or consists of the amino acid sequence of Subformula (XXVa): Q-X2-P-X4-X5-X6-P-Y-X9-V-V-X12-K-S, wherein:

    • X2 is F, I, L, N, S, or V;
    • X4 is E, G, L, R, W, or Y;
    • X5 is A, D, G, H, K, N, Q, S, T, V, or Y;
    • X6 is A, D, E, H, N, Q, S, or T;
    • X9 is E, F, H, N, V, or Y; and
    • X12 is E, H, I, K, or P.


In some embodiments, a peptide of Formula (XXV) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXVa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X6 is E; X9 is Y; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXV) and Subformula (XXVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is F, I, L, N, S, or V; but also, for example, those in which X2 is F; X2 is I; X2 is L; etc.; X2 is F or I; X2 is F, I, or L; X2 is I or L; X2 is I, L, N, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X5 or X6, etc.


Formula (XXVI)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXVI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is N, Q, R, S, or V;
    • X2 is A, D, F, H, I, L, N, or V;
    • X3 is L, P, Q, R, or S;
    • X4 is A, C, E, G, H, I, L, R, S, V, W, or Y;
    • X5 is E, G, K, N, P, Q, or S;
    • X6 is A, D, E, G, K, L, or R;
    • X7 is H, L, Q, S, or T;
    • X8 is F, H, K, or Y;
    • X9 is D, F, I, L, N, or Y;
    • X10 is A, F, G, L, M, or V;
    • X11 is A, F, G, I, L, M, P, V, or W;
    • X12 is A, K, N, T, or Y;
    • X13 is H, K, L, M, N, Q, S, T, or Y; and
    • X14 is C, E, G, N, P, or S.


In some embodiments, a peptide of Formula (XXVI) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 34, 38, 64, 127, 155, 156, 164, 213, 304, 329, 366, 371, 444, 445, 447, 493, and 555.


In some embodiments, a peptide of Formula XXVI comprises or consists of the amino acid sequence of Subformula (XXVIa): Q-X2-X3-X4-X5-X6-Q-Y-X9-V-X11-N-X13-S, wherein:

    • X2 is A, D, F, H, I, L, N, or V;
    • X3 is L, P, Q, R, or S;
    • X4 is A, C, E, G, H, I, L, R, S, V, W, or Y;
    • X5 is E, G, K, N, P, Q, or S;
    • X6 is A, D, E, G, K, L, or R;
    • X9 is D, F, I, L, N, or Y;
    • X11 is A, F, G, I, L, M, P, V, or W; and
    • X13 is H, K, L, M, N, Q, S, T, or Y.


In some embodiments, a peptide of Formula (XXVI) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXVIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X9 is Y; X11 is V; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXVI) and Subformula (XXVIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is L, P, Q, R, or S; but also, for example, those in which X3 is L; X3 is P; X3 is Q; etc.; X3 is L or P; X3 is L, P, or Q; X3 is P or Q; X3 is P, Q, R, or S, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X11 or X13, etc.


Formula (XXVII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXVII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, Q, or R;
    • X2 is F, G, I, L, or V;
    • X3 is A, H, P, Q, R, or S;
    • X4 is C, L, R, S, T, W, or Y;
    • X5 is A, E, G, H, P, Q, or V;
    • X6 is A, D, E, G, K, Q, or T;
    • X7 is A, H, Q, R, or S;
    • X8 is C, E, F, N, Q, S, or Y;
    • X9 is C, H, S, V, W, or Y;
    • X10 is D, I, L, T, or V;
    • X11 is A, E, G, or V;
    • X12 is K, P, or T;
    • X13 is K, M, N, Q, R, S, or T; and
    • X14 is S or Y.


In some embodiments, a peptide of Formula (XXVII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 53, 62, 204, 212, 275, 364, 374, 399, 437, 469, 474, 500, 524, 529, 533, and 548.


In some embodiments, a peptide of Formula (XXVII) comprises or consists of the amino acid sequence of Subformula (XXVIIa): Q-F-P-X4-E-X6-S-X8-Y-X10-V-K-X13-S, wherein:

    • X4 is C, L, R, S, T, W, or Y;
    • X6 is A, D, E, G, K, Q, or T;
    • X8 is C, E, F, N, Q, S, or Y;
    • X10 is D, I, L, T, or V; and
    • X13 is K, M, N, Q, R, S, or T.


In some embodiments, a peptide of Formula (XXVII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXVIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X6 is E; X8 is Y; X10 is V; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXVII) and Subformula (XXVIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is C, L, R, S, T, W, or Y; but also, for example, those in which X4 is C; X4 is L; X4 is R; etc.; X4 is C or L; X4 is C, L, or R; X4 is L or R; X4 is C, L, R, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X8 or X10, etc.


Formula (XXVIII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXVIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is H, Q, or S;
    • X2 is C, F, G, I, V, or Y;
    • X3 is A, P, Q, R, S, T, or Y;
    • X4 is C, L, N, S, W, or Y;
    • X5 is D, E, H, K, Q, S, or V;
    • X6 is A, E, G, K, or Q;
    • X7 is L or S;
    • X8 is C, N, P, S, V, or Y;
    • X9 is A, D, F, G, N, S, or Y;
    • X10 is D, G, L, S, V, or Y;
    • X11 is A, E, F, G, L, Q, S, or V;
    • X12 is A, K, N, Q, R, S, or T;
    • X13 is I, K, or T; and
    • X14 is I, L, P, R, S, T, or W.


In some embodiments, a peptide of Formula (XXVIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 121, 128, 136, 186, 200, 256, 261, 331, 345, 386, 389, 404, 441, and 450.


In some embodiments, a peptide of Formula (XXVIII) comprises or consists of the amino acid sequence of Subformula (XXVIIIa): Q-X2-X3-X4-X5-X6-S-X8-X9-X10-X11-X12-K-S, wherein:

    • X2 is C, F, G, I, V, or Y;
    • X3 is A, P, Q, R, S, T, or Y;
    • X4 is C, L, N, S, W, or Y;
    • X5 is D, E, H, K, Q, S, or V;
    • X6 is A, E, G, K, or Q;
    • X8 is C, N, P, S, V, or Y;
    • X9 is A, D, F, G, N, S, or Y;
    • X10 is D, G, L, S, V, or Y;
    • X11 is A, E, F, G, L, Q, S, or V; and
    • X12 is A, K, N, Q, R, S, or T.


In some embodiments, a peptide of Formula (XXVIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXVIIIa) does not comprise or consist of the amino acid sequence, wherein X2 is V, X3 is P, X4 is W; X5 is E; X6 is E; X8 is Y; X9 is Y; X10 is V; X11 is V; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXVIII) and Subformula (XXVIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is C, L, N, S, W, or Y; but also, for example, those in which X4 is C; X4 is L; X4 is N; etc.; X4 is C or L; X4 is C, L, or W; X4 is L or W; X4 is L, N, S, or Y, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X8 or X10, etc.


Formula (XXIX)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXIX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D or E;
    • X2 is F, G, H, I, R, or V;
    • X3 is A, L, P, or T;
    • X4 is F, G, L, R, S, T, W, or Y;
    • X5 is A, D, E, G, I, P, R, or V;
    • X6 is A, D, E, H, K, L, or R;
    • X7 is L, P, Q, R, S, T, or W;
    • X8 is F, H, N, R, S, V, or Y;
    • X9 is F, K, or Y;
    • X10 is A, D, F, G, I, L, N, R, or V;
    • X11 is A, E, F, G, L, M, or V;
    • X12 is E, K, P, Q, S, T, or V;
    • X13 is D, F, H, I, K, or Q; and
    • X14 is G, P, S, T, or Y.


In some embodiments, a peptide of Formula (XXIX) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 13, 108, 109, 139, 167, 183, 198, 205, 247, 285, 383, 440, 488, 502, and 546.


In some embodiments, a peptide of Formula (XXIX) comprises or consists of the amino acid sequence of Subformula (XXIXa): D-V-X3-X4-X5-X6-X7-X8-Y-X10-X11-X12-X13-X14, wherein:

    • X3 is A, L, P, or T;
    • X4 is F, G, L, R, S, T, W, or Y;
    • X5 is A, D, E, G, I, P, R, or V;
    • X6 is A, D, E, H, K, L, or R;
    • X7 is L, P, Q, R, S, T, or W;
    • X8 is F, H, N, R, S, V, or Y;
    • X10 is A, D, F, G, I, L, N, R, or V;
    • X11 is A, E, F, G, L, M, or V;
    • X12 is E, K, P, Q, S, T, or V;
    • X13 is D, F, H, I, K, or Q; and
    • X14 is G, P, S, T, or Y.


In some embodiments, a peptide of Formula (XXIX) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXIXa) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXIX) and Subformula (XXIXa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is F, G, L, R, S, T, W, or Y; but also, for example, those in which X4 is F; X4 is G; X4 is L; etc.; X4 is F or G; X4 is F, G, or L; X4 is G or L; X4 is G, L, R, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X7 or X8, etc.


Formula (XXX)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, H, L, or R;
    • X2 is F, G, H, I, L, S, or V;
    • X3 is L, P, or Q;
    • X4 is A, C, G, L, P, Q, R, S, T, V, or W;
    • X5 is D, E, G, H, K, or Q;
    • X6 is D, E, K, T, V, or Y;
    • X7 is A, E, L, P, R, S, T, or W;
    • X8 is D, E, F, H, S, W, or Y;
    • X9 is I, W, or Y;
    • X10 is A, D, I, P, or V;
    • X11 is A, C, F, G, H, I, L, M, T, or V;
    • X12 is D, E, H, K, T, or V;
    • X13 is G, H, I, K, L, N, Q, R, S, V, or Y; and
    • X14 is A, F, H, I, M, S, or Y.


In some embodiments, a peptide of Formula (XXX) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 21, 85, 95, 102, 112, 116, 129, 253, 259, 273, 327, 400, 423, 484, 486, 523, 526, and 539.


In some embodiments, a peptide of Formula (XXX) comprises or consists of the amino acid sequence of Subformula (XXXa): H-X2-P-X4-X5-E-X7-X8-Y-V-X11-K-X13-S, wherein:

    • X2 is F, G, H, I, L, S, or V;
    • X4 is A, C, G, L, P, Q, R, S, T, V, or W;
    • X5 is D, E, G, H, K, or Q;
    • X7 is A, E, L, P, R, S, T, or W;
    • X8 is D, E, F, H, S, W, or Y;
    • X11 is A, C, F, G, H, I, L, M, T, or V; and
    • X13 is G, H, I, K, L, N, Q, R, S, V, or Y.


In some embodiments, a peptide of Formula (XXX) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X5 is E; X7 is P; X8 is Y; X11 is V; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXX) and Subformula (XXXa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is F, G, H, I, L, S, or V; but also, for example, those in which X2 is F; X2 is G; X2 is H; etc.; X2 is F or G; X2 is F, G, or H; X2 is G or H; X2 is G, H, L, or V, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X7 or X8, etc.


Formula (XXXI)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, H, L, Q, R, T, or Y;
    • X2 is F, G, I, R, S, or V;
    • X3 is A, G, K, L, P, R, S, or T;
    • X4 is A, G, L, R, S, T, W, or Y;
    • X5 is A, E, G, H, K, N, P, Q, T, V, or Y;
    • X6 is A, D, E, G, or N;
    • X7 is A, P, Q, S, or T;
    • X8 is D, F, G, H, N, R, S, or Y;
    • X9 is D, F, G, H, K, N, Q, R, S, T, V, or Y;
    • X10 is E, I, L, N, T, V, or Y;
    • X11 is A, F, G, K, L, M, Q, or V;
    • X12 is I, K, L, N, Q, R, or T;
    • X13 is E, H, I, K, M, N, Q, T, or W; and
    • X14 is F, H, I, L, N, P, R, or S.


In some embodiments, a peptide of Formula (XXXI) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 18, 22, 23, 24, 49, 90, 111, 119, 131, 157, 222, 242, 264, 269, 301, 306, 344, 346, 347, 368, 373, 385, 414, 421, 429, and 492.


In some embodiments, a peptide of Formula (XXXI) peptide comprises or consists of the amino acid sequence of Subformula (XXXIa): X1-V-S-X4-X5-E-P-X8-X9-V-L-X12-X13-X14, wherein:

    • X1 is D, E, H, L, Q, R, T, or Y;
    • X4 is A, G, L, R, S, T, W, or Y;
    • X5 is A, E, G, H, K, N, P, Q, T, V, or Y;
    • X8 is D, F, G, H, N, R, S, or Y;
    • X9 is D, F, G, H, K, N, Q, R, S, T, V, or Y;
    • X12 is I, K, L, N, Q, R, or T;
    • X13 is E, H, I, K, M, N, Q, T, or W; and
    • X14 is F, H, I, L, N, P, R, or S.


In some embodiments, a peptide of Formula (XXXI) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXIa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X4 is W; X5 is E; X8 is Y; X9 is Y; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXI) and Subformula (XXXIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X12 is I, K, L, N, Q, R, or T; but also, for example, those in which X12 is I; X12 is K; X12 is L; etc.; X12 is I or K; X12 is I, K, or L; X12 is K or L; X12 is K, L, N, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X12, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1, X13, or X14, etc.


Formula (XXXII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, H, L, N, Q, or Y;
    • X2 is C, D, F, H, I, L, R, S, T, V, or Y;
    • X3 is A, G, L, N, P, Q, R, T, V, or Y;
    • X4 is C, F, G, R, S, or W;
    • X5 is A, E, or T;
    • X6 is A, D, E, Q, S, or Y;
    • X7 is A, L, P, T, or Y;
    • X8 is C, E, F, G, N, Q, S, or Y;
    • X9 is H, S, or Y;
    • X10 is A, F, H, L, R, S, T, or V;
    • X11 is A, C, D, E, F, G, H, L, M, N, Q, or V;
    • X12 is A, D, E, G, H, K, N, Q, S, or T;
    • X13 is D, E, F, I, K, L, M, N, P, Q, R, S, T, or Y; and
    • X14 is A, F, K, L, P, S, V, or Y.


In some embodiments, a peptide of Formula (XXXII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 10, 20, 41, 84, 97, 113, 122, 123, 130, 148, 165, 254, 277, 295, 297, 308, 342, 358, 432, 458, 461, 465, 514, 537, 540, 541, and 556.


In some embodiments, peptide of Formula (XXXII) comprises or consists of the amino acid sequence of Subformula (XXXIIa): Q-X2-X3-X4-E-E-P-Y-Y-V-X11-K-X13-X14, wherein:

    • X2 is C, D, F, H, I, L, R, S, T, V, or Y;
    • X3 is A, G, L, N, P, Q, R, T, V, or Y;
    • X4 is C, F, G, R, S, or W;
    • X11 is A, C, D, E, F, G, H, L, M, N, Q, or V;
    • X13 is D, E, F, I, K, L, M, N, P, Q, R, S, T, or Y; and
    • X14 is A, F, K, L, P, S, V, or Y.


In some embodiments, a peptide of Subformula (XXXII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Formula (XXXIIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X4 is W; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXII) and Subformula (XXXIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is X4 is C, F, G, R, S, or W; but also, for example, those in which X4 is C; X1 is F; X4 is G; etc.; X4 is C or F; X4 is C, F, or G; X4 is F or G; X4 is F, R, S, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X1 or X3, etc.


Formula (XXXIII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, H, L, N, Q, R, or Y;
    • X2 is A, D, E, F, I, L, T, V, or Y;
    • X3 is A, E, L, P, Q, R, or S;
    • X4 is F, G, P, R, S, or W;
    • X5 is A, D, E, G, H, Q, S, or V;
    • X6 is A, D, E, G, H, Q, or T;
    • X7 is P, Q, R, S, or T;
    • X8 is D, F, H, K, N, S, or Y;
    • X9 is D, F, G, H, N, P, Q, S, or Y;
    • X10 is A, D, F, G, I, L, P, R, or V;
    • X11 is A, E, G, I, L, R, or V;
    • X12 is D, E, K, N, Q, R, T, or V;
    • X13 is D, E, I, K, L, Q, R, or T; and
    • X14 is A, I, S, T, or Y.


In some embodiments, a peptide of Formula (XXXIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 5, 33, 71, 79, 106, 120, 151, 168, 219, 226, 229, 237, 268, 326, 330, 334, 375, 392, 395, 426, 433, 434, 472, 490, 528, and 544.


In some embodiments, the peptide comprises or consists of the amino acid sequence of Subformula (XXXIIIa): Q-X2-X3-W-E-X6-P-X8-Y-X10-X11-X12-K-S, wherein:

    • X2 is A, D, E, F, I, L, T, V, or Y;
    • X3 is A, E, L, P, Q, R, or S;
    • X6 is A, D, E, G, H, Q, or T;
    • X8 is D, F, H, K, N, S, or Y;
    • X10 is A, D, F, G, I, L, P, R, or V;
    • X11 is A, E, G, I, L, R, or V; and
    • X12 is D, E, K, N, Q, R, T, or V.


In some embodiments, a peptide of Formula (XXXIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXIIIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X6 is E; X8 is Y; X10 is V; X11 is V; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXIII) and Subformula (XXXIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, D, E, F, I, L, T, V, or Y; but also, for example, those in which X2 is A; X2 is D; X2 is E; etc.; X2 is A or D; X2 is D, E, or F; X2 is D or E; X2 is D, E, F, or T, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X10 or X11, etc.


Formula (XXXIV)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXIV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, D, E, K, L, N, Q, R, S, T, or Y;
    • X2 is D, E, F, I, L, N, Q, R, S, T, or V;
    • X3 is I, L, P, Q, or T;
    • X4 is F, G, I, K, M, N, Q, R, S, V, W, or Y;
    • X5 is A, D, E, G, P, Q, V, or Y;
    • X6 is A, D, E, G, H, L, P, Q, S, or V;
    • X7 is F or P;
    • X8 is D, G, K, N, R, S, V, or Y;
    • X9 is D, E, L, or Y;
    • X10 is D, F, G, K, L, M, N, R, or V;
    • X11 is A, E, F, G, M, P, Q, R, or V;
    • X12 is K, L, N, or V;
    • X13 is A, D, K, M, N, T, or V; and
    • X14 is A, F, H, L, P, S, T, V, or Y.


In some embodiments, a peptide of Formula (XXXIV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 14, 43, 44, 104, 132, 145, 160, 191, 210, 221, 284, 323, 324, 328, 353, 372, 387, 391, 411, 431, 435, 468, 471, 481, 509, 510, 516, 525, and 547.


In some embodiments, a peptide of Formula (XXXIV) or consists of the amino acid sequence of Subformula (XXXIVa): X1-X2-P-X4-X5-X6-P-X8-Y-X10-V-X12-N-S, wherein:

    • X1 is A, D, E, K, L, N, Q, R, S, T, or Y;
    • X2 is D, E, F, I, L, N, Q, R, S, T, or V;
    • X4 is F, G, I, K, M, N, Q, R, S, V, W, or Y;
    • X5 is A, D, E, G, P, Q, V, or Y;
    • X6 is A, D, E, G, H, L, P, Q, S, or V;
    • X8 is D, G, K, N, R, S, V, or Y;
    • X10 is D, F, G, K, L, M, N, R, or V; and
    • X12 is K, L, N, or V.


In some embodiments, a peptide of Formula (XXXIV) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXIVa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X4 is W; X5 is E; X6 is E; X8 is Y; X10 is V; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXIV) and Subformula (XXXIVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X1 is A, D, E, K, L, N, Q, R, S, T, or Y; but also, for example, those in which X1 is A; X1 is D; X1 is E; etc.; X1 is A or D; X1 is A, D, or E; X1 is D or E; X1 is D, E, Q, or R, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X1, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X5 or X6, etc.


Formula (XXXV)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, E, H, L, N, Q, R, S, V, or Y;
    • X2 is F, I, N, S, V, or Y;
    • X3 is A, E, I, L, P, R, S, T, V, W, or Y;
    • X4 is C, F, G, K, L, R, S, W, or Y;
    • X5 is D, E, I, K, Q, V, or Y;
    • X6 is A, D, E, G, N, P, Q, R, S, V, or Y;
    • X7 is G, L, P, Q, or S;
    • X8 is H, I, L, N, S, or Y;
    • X9 is A, D, E, F, L, N, S, T, V, or Y;
    • X10 is A, D, E, F, G, H, I, L, N, S, T, or V;
    • X11 is K, L, M, or V;
    • X12 is I, K, N, P, Q, R, S, or T;
    • X13 is A, C, E, H, K, M, N, Q, R, S, T, or Y; and
    • X14 is F, H, P, S, T, or V.


In some embodiments, a peptide of Formula (XXXV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 25, 37, 68, 83, 101, 124, 147, 153, 158, 159, 182, 209, 232, 233, 235, 248, 262, 270, 283, 302, 384, 390, 401, 407, 409, 418, 448, 457, 515, 519, and 530.


In some embodiments, a peptide of Formula (XXXV) comprises or consists of the amino acid sequence of Subformula (XXXVa): X1-X2-X3-X4-E-X6-P-Y-Y-X10-V-X12-X13-S, wherein:

    • X1 is A, E, H, L, N, Q, R, S, V, or Y;
    • X2 is F, I, N, S, V, or Y;
    • X3 is A, E, I, L, P, R, S, T, V, W, or Y;
    • X4 is C, F, G, K, L, R, S, W, or Y;
    • X6 is A, D, E, G, N, P, Q, R, S, V, or Y;
    • X10 is A, D, E, F, G, H, I, L, N, S, T, or V;
    • X12 is I, K, N, P, Q, R, S, or T; and
    • X13 is A, C, E, H, K, M, N, Q, R, S, T, or Y.


In some embodiments, a peptide of Formula (XXXV) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXVa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X6 is E; X10 is V; X12 is K; and X13 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXV) and Subformula (XXXVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is F, I, N, S, V, or Y; but also, for example, those in which X2 is F; X2 is I; X2 is N; etc.; X2 is F or I; X2 is F, I, or N; X2 is I or N; X2 is I, N, V, or Y, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X4 or X6, etc.


Formula (XXXVI)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXVI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, F, H, L, N, Q, R, S, T, or Y;
    • X2 is A, D, F, G, I, L, Q, or V;
    • X3 is A, D, E, F, G, H, K, L, P, Q, R, S, T, or V;
    • X4 is A, C, E, F, G, I, L, P, Q, R, S, V, W, or Y;
    • X5 is A, D, E, G, H, I, K, L, N, P, Q, R, S, T, V, or Y;
    • X6 is A, D, E, G, K, N, P, Q, S, T, V, or Y;
    • X7 is A, H, I, L, P, Q, R, S, or T;
    • X8 is A, E, F, H, I, K, L, P, Q, R, S, V, or Y;
    • X9 is A, C, D, F, G, H, K, L, N, P, Q, R, S, or Y;
    • X10 is A, D, E, F, G, H, I, L, N, P, S, V, or Y;
    • X11 is A, D, E, F, G, I, L, M, N, S, T, or V;
    • X12 is D, E, G, H, I, K, N, Q, R, S, T, W, or Y;
    • X13 is D, E, H, K, N, Q, R, S, T, or V; and
    • X14 is A, I, P, S, or Y.


In some embodiments, a peptide of Formula (XXXVI) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 15, 67, 69, 75, 81, 86, 96, 99, 107, 126, 135, 140, 142, 144, 149, 170, 173, 175, 178, 188, 192, 196, 197, 206, 207, 217, 223, 231, 234, 239, 249, 250, 251, 257, 271, 272, 287, 291, 307, 314, 315, 317, 318, 319, 320, 349, 350, 359, 360, 362, 363, 365, 370, 378, 382, 406, 410, 427, 438, 456, 476, 485, 506, 512, 520, 527, 531, 538, and 545.


In some embodiments, a peptide of Formula (XXXVI) comprises or consists of the amino acid sequence of Subformula (XXXVIa): H-V-X3-X4-X5-E-P-Y-X9-X10-V-X12-K-S, wherein:

    • X3 is A, D, E, F, G, H, K, L, P, Q, R, S, T, or V;
    • X4 is A, C, E, F, G, I, L, P, Q, R, S, V, W, or Y;
    • X5 is A, D, E, G, H, I, K, L, N, P, Q, R, S, T, V, or Y;
    • X9 is A, C, D, F, G, H, K, L, N, P, Q, R, S, or Y;
    • X10 is A, D, E, F, G, H, I, L, N, P, S, V, or Y; and
    • X12 is D, E, G, H, I, K, N, Q, R, S, T, W, or Y.


In some embodiments, a peptide of Formula (XXXVI) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXVIa) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X9 is Y; X10 is V; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXVI) and Subformula (XXXVIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is A, D, E, F, G, H, K, L, P, Q, R, S, T, or V; but also, for example, those in which X3 is A; X3 is D; X3 is F; etc.; X3 is A or D; X3 is A, D, or E; X3 is D or E; X3 is D, E, F, or P, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X9 or X10, etc.


Formula (XXXVII)

In some embodiments, the peptide comprises or consists of the amino acid sequence of Formula (XXXVII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, E, H, Q, or R;
    • X2 is D, F, G, L, M, or V;
    • X3 is A, E, H, P, Q, R, S, T, or W;
    • X4 is C, G, L, R, S, V, or W;
    • X5 is A, D, E, G, I, K, L, or Y;
    • X6 is A, D, E, G, K, L, N, Q, S, or V;
    • X7 is A, H, L, M, P, S, T, or W;
    • X8 is C, H, I, L, N, S, V, or Y;
    • X9 is D, H, L, N, S, or Y;
    • X10 is A, D, E, F, G, L, P, S, T, or V;
    • X11 is A, D, E, G, I, M, Q, R, T, V, or W;
    • X12 is H, I, K, L, N, Q, R, or T;
    • X13 is A, E, H, I, K, M, N, R, or V; and
    • X14 is F, P, S, or T.


In some embodiments, a peptide of Formula (XXXVII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 28, 59, 60, 78, 91, 133, 187, 199, 241, 243, 281, 282, 290, 416, 422, 439, 451, 454, 459, 460, 495, 503, 513, 551, and 552.


In some embodiments, a peptide of Formula (XXXVII) comprises or consists of the amino acid sequence of Subformula (XXXVIIa): H-V-X3-X4-X5-X6-X7-Y-Y-X10-X11-K-N-S, wherein:

    • X3 is A, E, H, P, Q, R, S, T, or W;
    • X4 is C, G, L, R, S, V, or W;
    • X5 is A, D, E, G, I, K, L, or Y;
    • X6 is A, D, E, G, K, L, N, Q, S, or V;
    • X7 is A, H, L, M, P, S, T, or W;
    • X10 is A, D, E, F, G, L, P, S, T, or V; and
    • X11 is A, D, E, G, I, M, Q, R, T, V, or W.


In some embodiments, a peptide of Formula (XXXVII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXVIIa) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X10 is V; and X11 is V.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXVII) and Subformula (XXXVIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is A, E, H, P, Q, R, S, T, or W; but also, for example, those in which X3 is D; X1 is H; X3 is L; etc.; X3 is D or H; X3 is D, H, or L; X3 is H or L; X3 is H, L, N, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X10 or X11, etc.


Formula (XXXVIII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XXXVIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is D, E, G, H, K, L, R, or S;
    • X2 is A, F, I, or L;
    • X3 is A, E, F, L, P, Q, R, or S;
    • X4 is E, F, G, M, R, S, V, W, or Y;
    • X5 is E, P, or Q;
    • X6 is A, D, E, or G;
    • X7 is H, L, P, R, S, or T;
    • X8 G, L, S, or Y;
    • X9 is D, H, K, L, T, or Y;
    • X10 is A, F, G, I, L, or R;
    • X11 L, P, or V;
    • X12 is A, E, H, I, K, or R;
    • X13 is E, I, K, N, Q, R, S, or T; and
    • X14 is A, F, H, P, R, S, T, or Y.


In some embodiments, a peptide of Formula (XXXVIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 17, 26, 114, 174, 179, 184, 195, 309, 322, 325, 348, 417, 462, 464, 494, 505, and 518.


In some embodiments, a peptide of Formula (XXXVIII) comprises or consists of the amino acid sequence of Subformula (XXXVIIIa): H-L-P-X4-E-E-X7-Y-X9-X10-V-X12-X13-X14, wherein:

    • X4 is E, F, G, M, R, S, V, W, or Y;
    • X7 is H, L, P, R, S, or T;
    • X9 is D, H, K, L, T, or Y;
    • X12 is A, E, H, I, K, or R;
    • X13 is E, I, K, N, Q, R, S, or T; and
    • X14 is A, F, H, P, R, S, T, or Y.


In some embodiments, a peptide of Formula (XXXVIII) does not comprise or consist of the amino acid sequence, wherein X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXVIIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X7 is P; X9 is Y; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXVIII) and Subformula (XXXVIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is E, F, G, M, R, S, V, W, or Y; but also, for example, those in which X4 is E; X4 is F; X4 is M; etc.; X4 is E or F; X4 is E, F, or G; X4 is F or G; X4 is F, G, M, or R, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X13 or X14, etc.


Formula (XXXIX)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XXXIX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is A, E, N, Q, R, or W;
    • X2 is A, D, I, L, S, T, or V;
    • X3 is E, H, L, P, T, or Y;
    • X4 is L, S, or W;
    • X5 is A, E, G, V, or Y;
    • X6 is E, G, I, K, P, Q, or T;
    • X7 is H, P, R, or S;
    • X8 is S or Y;
    • X9 is F, S, W, or Y;
    • X10 is A, D, F, G, H, I, T, or V;
    • X11 is L or V;
    • X12 is E, H, K, or N;
    • X13 is H, K, N, or V; and
    • X14 is F, S, T, or Y.


In some embodiments, a peptide of Formula (XXXIX) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 8, 52, 89, 177, 189, 225, 279, 335, 379, 408, 424, 521, and 535.


In some embodiments, peptide of Formula (XXXIX) comprises or consists of the amino acid sequence of Subformula (XXXIXa): Q-X2-X3-W-X5-X6-P-Y-Y-X10-V-K-K-X14, wherein:

    • X2 is A, D, I, L, S, T, or V;
    • X3 is E, H, L, P, T, or Y;
    • X5 is A, E, G, V, or Y;
    • X6 is E, G, I, K, P, Q, or T;
    • X10 is A, D, F, G, H, I, T, or V; and
    • X14 is F, S, T, or Y.


In some embodiments, a peptide of Formula (XXXIX) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XXXIXa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X5 is E; X6 is E; X10 is V; and X14 is S. More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XXXIX) and Subformula (XXXIXa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, D, I, L, S, T, or V; but also, for example, those in which X2 is A; X2 is D; X2 is I; etc.; X2 is A or D; X2 is A, D, or I; X2 is D or I; X2 is D, I, L, or T, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X5 or X6, etc.


Formula (XL)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XL): X1-X2-X3-X4-X5-X6-X7-X8-X9-V-X11-X12-X13-X14, wherein:

    • X1 is D, E, H, L, or Q;
    • X2 is D, F, L, S, or V;
    • X3 is A, H, L, P, Q, or S;
    • X4 is E, G, M, R, or W;
    • X5 is A, D, E, Q, or T;
    • X6 is D or E;
    • X7 is A, L, R, S, or V;
    • X8 is F, L, N, or Y;
    • X9 is F, G, H, L, N, S, or Y;
    • X11 is K, M, or V;
    • X12 is K or T;
    • X13 is D, K, or N; and
    • X14 is F, P, S, or T.


In some embodiments, a peptide of Formula (XL) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 32, 40, 51, 65, 134, 152, 185, 216, 258, 367, and 388.


In some embodiments, the peptide comprises or consists of the amino acid sequence of Subformula (XLa): X1-X2-X3-W-X5-E-X7-X8-X9-V-V-K-N-X14, wherein:

    • X1 is D, E, H, L, or Q;
    • X2 is D, F, L, S, or V;
    • X3 is A, H, L, P, Q, or S;
    • X5 is A, D, E, Q, or T;
    • X7 is A, L, R, S, or V;
    • X8 is F, L, N, or Y;
    • X9 is F, G, H, L, N, S, or Y; and
    • X14 is F, P, S, or T.


In some embodiments, a peptide of Formula (XL) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X8 is Y; X9 is Y; Xu is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XLa) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X5 is E; X8 is Y; X9 is Y; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XL) and Subformula (XLa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X3 is A, H, L, P, Q, or S; but also, for example, those in which X3 is A; X3 is H; X3 is L; etc.; X3 is A or H; X3 is A, H, or L; X3 is H or L; X3 is H, L, P, or Q, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X3, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X7 or X8, etc.


Formula (XLI)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XL1): X1-X2-X3-X4-X5-X6-X7-X8-X9-V-X11-X12-X13-X14, wherein:

    • X1 is H or Q;
    • X2 is A, F, I, L, T, or V;
    • X3 is H, L, P, R, S, or T;
    • X4 is A, F, G, L, S, W, or Y;
    • X5 is A, D, E, G, N, or T;
    • X6 is A, E, or V;
    • X7 is A, H, P, S, or T;
    • X8 is A, D, H, N, Q, T, or Y;
    • X9 is G, H, L, S, T, V, or Y;
    • X11 is S or V;
    • X12 is K, L, N, P, Q, R, T, or Y;
    • X13 is E, H, I, K, N, or T; and
    • X14 is F or S.


In some embodiments, a peptide of Formula (XLI) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 39, 55, 61, 73, 110, 137, 161, 171, 190, 230, 341, 380, 489, and 543.


In some embodiments, a peptide of Formula (XLI) comprises or consists of the amino acid sequence of Subformula (XLIa): Q-X2-X3-X4-X5-E-X7-X8-X9-V-V-X12-K-S, wherein:

    • X2 is A, F, I, L, T, or V;
    • X3 is H, L, P, R, S, or T;
    • X4 is A, F, G, L, S, W, or Y;
    • X5 is A, D, E, G, N, or T;
    • X7 is A, H, P, S, or T;
    • X8 is A, D, H, N, Q, T, or Y;
    • X9 is G, H, L, S, T, V, or Y; and
    • X12 is K, L, N, P, Q, R, T, or Y.


In some embodiments, a peptide of Formula (XLI) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X5 is E; X6 is E; X7 is P; X8 is Y; X9 is Y; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XLIa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X3 is P; X4 is W; X5 is E; X7 is P; X8 is Y; X9 is Y; and X12 is K.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XLI) and Subformula (XLIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X2 is A, F, I, L, T, or V; but also, for example, those in which X2 is A; X2 is F; X2 is I; etc.; X2 is A or F; X2 is A, F, or F; X2 is F or I; X2 is A, F, I, or T, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X2, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X4 or X5, etc.


Formula (XLII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XLII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X1 is H, L, Q, R, or V;
    • X2 is F, G, H, I, L, or V;
    • X3 is A, H, N, P, Q, or T;
    • X4 is F, G, L, M, S, T, or W;
    • X5 is A, D, G, I, K, P, Q, or V;
    • X6 is D, E, G, K, or P;
    • X7 is A, L, P, Q, R, S, or T;
    • X8 is F, H, S, or Y;
    • X9 is A, D, F, G, H, K, N, S, or Y;
    • X10 is I, L, N, or V;
    • X11 is A, E, G, K, L, Q, S, V, or W;
    • X12 is A, D, E, G, I, K, M, N, Q, R, or T;
    • X13 is E, H, K, N, Q, R, T, or V; and
    • X14 is F, H, N, P, S, T, or Y.


In some embodiments, a peptide of Formula (XLII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 3, 4, 30, 36, 80, 94, 118, 181, 214, 224, 240, 260, 286, 288, 289, 312, 313, 357, 377, 393, 405, 504, and 550.


In some embodiments, a peptide of Formula (XLII) comprises or consists of the amino acid sequence of Subformula (XLIIa): H-V-P-X4-X5-G-X7-Y-X9-V-X11-X12-X13-X14, wherein:

    • X4 is F, G, L, M, S, T, or W;
    • X5 is A, D, G, I, K, P, Q, or V;
    • X7 is A, L, P, Q, R, S, or T;
    • X9 is A, D, F, G, H, K, N, S, or Y;
    • X11 is A, E, G, K, L, Q, S, V, or W;
    • X12 is A, D, E, G, I, K, M, N, Q, R, or T;
    • X13 is E, H, K, N, Q, R, T, or V; and
    • X14 is F, H, N, P, S, T, or Y.


In some embodiments, a peptide of Formula (XLII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XLIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X7 is P; X9 is Y; X11 is V; X12 is K; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XLII) and Subformula (XLIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is F, G, L, M, S, T, or W; but also, for example, those in which X4 is F; X4 is G; X4 is L; etc.; X4 is F or G; X4 is F, G, or L; X4 is G or L; X4 is G, L, M, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X11 or X12, etc.


Formula (XLIII)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XLIII): X1-X2-X3-X4-X5-X6-X7-Y-X9-V-X11-X12-X13-X14, wherein:

    • X1 is G, H, N, Q, R, or Y;
    • X2 is F, H, I, L, M, or V;
    • X3 is L, P, or Q;
    • X4 is G, L, Q, R, S, or W;
    • X5 is G, H, K, Q, R, or V;
    • X6 is E or V;
    • X7 is P or R;
    • X9 is N, S, Y;
    • X11 is L, Q, or V;
    • X12 is A, E, K, L, or T;
    • X13 is K, M, N, P, Q, or T; and
    • X14 is A, F, I, L, P, S, or Y.


In some embodiments, a peptide of Formula (XLIII) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 35, 76, 215, 252, 294, 305, 310, 311, 316, 355, 412, 413, 498, and 549.


In some embodiments, the peptide comprises or consists of the amino acid sequence of Subformula (XLIIIa): H-V-P-X4-Q-E-P-Y-Y-V-V-K-X13-X14, wherein:

    • X4 is G, L, Q, R, S, or W;
    • X13 is K, M, N, P, Q, or T; and
    • X14 is A, F, I, L, P, S, or Y.


In some embodiments, a peptide of Formula (XLIII) does not comprise or consist of the amino acid sequence, wherein X1 is Q; X2 is V; X3 is P; X4 is W; X6 is E; X7 is P; X9 is Y; X11 is V; X12 is K; X13 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XLIIIa) does not comprise or consist of the amino acid sequence, wherein X4 is W; X13 is K; and X14 is S.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XLIII) and Subformula (XLIIIa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is G, L, Q, R, S, or W; but also, for example, those in which X4 is G; X4 is L; X4 is Q; etc.; X4 is G or L; X4 is G, L, or Q; X4 is G or L; X4 is L, Q, R, or S, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X13 or X14, etc.


Formula (XLIV)

In some embodiments, a peptide comprises or consists of the amino acid sequence of Formula (XLIV): Q-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:

    • X2 is H, I, M, N, or V;
    • X3 is A, L, T, or V;
    • X4 is F, H, L, N, S, or W;
    • X5 is A, D, K, N, or S;
    • X6 is A, E, H, K, or Q;
    • X7 is P or S;
    • X8 is F, H, N, or Y;
    • X9 is N, S, T, or Y;
    • X10 is G, P, S, V, or Y;
    • X11 is Q or V;
    • X12 is E or K;
    • X13 is D, N, P, R, or T; and
    • X14 is S, T, or Y.


In some embodiments, a peptide of Formula (XLIV) comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 42, 54, 56, 154, 162, 180, 396, 467, and 483.


In some embodiments, a peptide of Formula (XLIV) comprises or consists of the amino acid sequence of Subformula (XLIVa): Q-X2-X3-X4-X5-X6-P-Y-X9-X10-V-K-X13-S, wherein:

    • X2 is H, I, M, N, or V;
    • X3 is A, L, T, or V;
    • X4 is F, H, L, N, S, or W;
    • X5 is A, D, K, N, or S;
    • X6 is A, E, H, K, or Q;
    • X9 is N, S, T, or Y;
    • X10 is G, P, S, V, or Y; and
    • X13 is D, N, P, R, or T.


In some embodiments, a peptide of Formula (XLIV) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X6 is E; X7 is P; X8 is Y; X9 is Y; X10 is V; X11 is V; X12 is K; and X14 is S.


In some embodiments, a peptide of Subformula (XLIVa) does not comprise or consist of the amino acid sequence, wherein X2 is V; X4 is W; X6 is E; X9 is Y; and X10 is V.


More generally, in some embodiments, the selection of amino acids at any position of a peptide of Formula (XLIV) and Subformula (XLIVa) can be any subset or combination of choices at that position. Thus, the embodiments not only include those in which X4 is F, H, L, N, S, or W; but also, for example, those in which X4 is F; X4 is H; X4 is L; etc.; X4 is F or H; X4 is F, H, or L; X4 is H or L; X4 is H, L, N, or W, and so on. Moreover, the choice of amino acids, or combination or subset thereof, at one position, e.g., X4, is independent of the choice of amino acids (or combination or subset thereof) at any other position, e.g., X9 or X10, etc.


In some embodiments, a nerve targeting peptide of the present disclosure shows selective nerve binding, including a peptide consisting or comprising the amino acid sequence of any one of SEQ ID NOS: 1-557 or a peptide consisting or comprising the amino acid sequence of any one of Formulas I-XLIV, including Subformulas Ia-XLIVa.


In some embodiments, a nerve targeting peptide of the present disclosure show selective binding to nerves versus muscle, including a peptide consisting or comprising an amino acid sequence of any one of SEQ ID NOS: 1-557 or a peptide consisting or comprising an amino acid sequence of any one of Formulas I-XLIV, including Subformulas Ia-XLIVa.


In some embodiments, a nerve targeting peptide of the present disclosure, including one comprising the amino acid sequence of Formulas (I)-(XLIV) or Subformulas (Ia)-(XLIVa), as applicable, does not include the amino acid sequence of SGQVPWEEPYYVVKKSS (HNP-401, SEQ ID NO:559); SGQVPWEEPYYVVKKSSGGC (HNP-401 with GGC linker; SEQ ID NO:560); QVPWEEPYYVVKKSS (HNP401-N-2; SEQ ID NO: 561); QVPWEEPYYVVKKSSGG (HNP401-N-2 with GG linker; SEQ ID NO: 562) and QVPWEEPYYVVKKSSGGC (HNP401-N-2 with GGC linker; SEQ ID NO: 563).


In some embodiments, peptides of the present disclosure are acetylated at the N-terminus (“Ac” or “acetyl”), amidated at the C-terminus (“CONH2” or “NH2”), or both.


The peptides of the present disclosure may be synthesized by any suitable method. In some embodiments, the peptides are chemically synthesized, e.g., by solid phase peptide synthesis. Techniques for solid phase synthesis are described, for example, by Barany and Merrifield (1963) Solid-Phase Peptide Synthesis; pp. 3-284 in The Peptides: Analysis, Synthesis, Biology. Vol. 2: Special Methods in Peptide Synthesis, Part A.; Merrifield et al. (1963) J. Am. Chem. Soc, 85: 2149-2156, and Stewart et al. (1984) Solid Phase Peptide Synthesis, 2nd ed. Pierce Chem. Co., Rockford, 111. In some embodiments, the peptides are recombinantly produced via cell-based expression systems. A polynucleotide encoding a peptide of the present disclosure may be incorporated into a recombinant expression vector for production of the peptide in a host cell. The expression vector also comprises expression control sequences, such as a promoter, enhancer, initiation site, and the like. The polynucleotide encoding the peptide is operatively linked to at least one expression control sequence in the expression vector to direct mRNA synthesis. Host cells may be genetically engineered (transduced, transformed, or transfected) with the vectors. The vector may be in the form of a plasmid, a viral particle, a phage, etc. The engineered host cells can be cultured in appropriate media and under suitable conditions, such as temperature, pH, and the like, and for a suitable time to express a sufficient amount of the peptide. In some embodiments, the synthesized peptides are purified using methods such as preparative reversed phase chromatography, partition chromatography, gel filtration, gel electrophoresis, or ion-exchange chromatography or other methods used in the art.


II. Cargo Molecule

In some embodiments, the peptide of a nerve targeting peptide conjugate is directly bound to a cargo molecule. In some embodiments, the peptide of a nerve targeting peptide conjugate is indirectly (e.g., via a linker) bound to a cargo molecule. In some embodiments, the peptide is bound to a cargo molecule at its N-terminus, at its C-terminus, or at an internal position (e.g., to an internal amino acid) of the peptide. In some embodiments, two, three, four or more peptides are directly or indirectly bound to a cargo molecule.


In certain embodiments, a cargo molecule comprises a drug, fluorescent moiety, photosensitizing agent, radiolabel, neurotrophic factor, or a combination thereof. In some embodiments, the cargo molecule comprises a drug. In some embodiments, the cargo molecule comprises a fluorescent moiety. In some embodiments, the cargo molecule comprises a photosensitizing agent. In some embodiments, the peptide is bound to two or more cargo molecules. The two or more cargo molecule may be the same molecule or different molecule, or be from the same class of cargo molecule (e.g., two drugs) or from different classes of cargo molecule (e.g., one drug and one fluorescent moiety).


In some embodiments, the peptide comprises or consists or the amino acid sequence set forth in any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:1-30. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:31-36. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:37-40. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:41-240. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:241-349. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:350-449. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:450-557.


In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of Formulas (I)-(XLIV) and Subformulas (Ia)-(XLIVa).


Fluorescent Moieties

In some embodiments, the cargo molecule of the nerve targeting peptide conjugate comprises a fluorescent moiety (e.g., a fluorescent protein, fluorescent peptide, or fluorophore (fluorescent dye) molecule). All fluorescent moieties are encompassed within the term “fluorescent moiety.” Specific examples of fluorescent moieties given herein, are illustrative and are not meant to limit the fluorescent moieties for use with the targeting molecules disclosed herein. Common classes of fluorophores (fluorescent dyes) include, but are not limited to, xanthenes, such as rhodamines, rhodols and fluoresceins, and their derivatives; bimanes; coumarins and their derivatives such as umbelliferone and aminomethyl coumarins; aromatic amines such as dansyl; squarate dyes; benzofurans; fluorescent cyanines; carbazoles; dicyanomethylene pyranes; polymethine; oxabenzanthrane; xanthene; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare-earth metal chelate complexes; and derivatives of such dyes. Fluorescent dyes are discussed, for example, in U.S. Pat. Nos. 4,452,720; 5,227,487; and 5,543,295.


In some embodiments, the fluorescent moiety is selected from the group consisting of a xanthene; a bimane; a coumarin; an aromatic amines; a benzofuran; a fluorescent cyanine; a carbazole; a dicyanomethylene pyrane; polymethine; oxabenzanthrane; pyrylium; carbostyl; perylene; acridone; quinacridone; rubrene; anthracene; coronene; phenanthrecene; pyrene; butadiene; stilbene; porphyrin; pthalocyanine; lanthanide metal chelate complexes; rare-earth metal chelate complexes; FITC; Cy3; EGFP; cyan fluorescent protein (CFP); EGFP; 5-FAM; 6-FAM; FAM; fluorescein, IAEDANS, EDANS and BODIPY FL; TRITC; Cy5; Cy3; YFP; 6-FAM; LC Red 640; Alexa Fluor 546; fluorescein; tetramethylrhodamine; Dabcyl; BODIPY FL; QSY 7, QSY 9, QSY 21 and BBQ-650 dyes.


In some embodiments, the cargo molecule comprises a fluorescein dye. Typical fluorescein dyes include, but are not limited to, 5-carboxyfluorescein, fluorescein-5-isothiocyanate, 5(6)-carboxyfluorescein, 5,6-dicarboxyfluorescein, 5-(and 6)-sulfofluorescein, sulfonefluorescein, succinyl fluorescein, 5-(and 6)-carboxy SNARF-1, carboxyfluorescein sulfonate, carboxyfluorescein zwitterion, carboxyfluorescein quaternary ammonium, carboxyfluorescein phosphonate, carboxyfluorescein GABA, carboxyfluorescein-cys-Cy5,5′(6′)-carboxyfluorescein, fluorescein glutathione, and 6-carboxyfluorescein; examples of other fluorescein dyes can be found, for example, in U.S. Pat. Nos. 6,008,379, 5,750,409, 5,066,580, and 4,439,356.


In some embodiments, the cargo molecule comprises a rhodamine dye, such as, for example, 5-(and 6)-carboxy rhodamine 110, tetramethylrhodamine-6-isothiocyanate, 5-carboxytetramethylrhodamine, 5-carboxy rhodol derivatives, tetramethyl and tetraethyl rhodamine, diphenyldimethyl and diphenyldiethyl rhodamine, dinaphthyl rhodamine, rhodamine 101 sulfonyl chloride (sold under the tradename of TEXAS RED®), and other rhodamine dyes. Other rhodamine dyes can be found, for example, in U.S. Pat. Nos. 6,080,852; 6,025,505; 5,936,087; 5,750,409. In some embodiments, a cargo moiety includes a cyanine dye, such as, for example, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy 7, and indocyanine green.


In some embodiments, the fluorophore exhibits green fluorescence (such as for example 494 nm/519 nm), orange fluorescence (such as for example 554 nm/570 nm), red fluorescence (such as for example 590 nm/617 nm), or far red fluorescence (such as for example 651 nm/672 nm) excitation/emission spectra. In some embodiments, the fluorophore is a fluorophore with excitation and emission spectra in the range of about 350 nm to about 775 nm. In some embodiments the excitation and emission spectra are about 346 nm/446 nm, about 494 nm/519 nm, about 554 nm/570 nm, about 555 nm/572 nm, about 590 nm/617 nm, about 651 nm/672 nm, about 679 nm/702 nm or about 749 nm/775 nm.


In some embodiments, the fluorophore can include but is not limited to AlexaFluor 3, AlexaFluor 5, AlexaFluor 350, AlexaFluor 405, AlexaFluor 430, AlexaFluor 488, AlexaFluor 500, AlexaFluor 514, AlexaFluor 532, AlexaFluor 546, AlexaFluor 555, AlexaFluor 568, AlexaFluor 594, AlexaFluor 610, AlexaFluor 633, AlexaFluor 647, AlexaFluor 660, AlexaFluor 680, AlexaFluor 700, and AlexaFluor 750 (Molecular Probes AlexaFluor dyes, available from Life Technologies, Inc. (USA)). In some embodiments, the fluorophore can include but is not limited to Cy dyes, including Cy2, Cy3, Cy3B, Cy3.5, Cy5, Cy5.5 and Cy7 (available from GE Life Sciences or Lumiprobes). In some embodiments the fluorophore can include but is not limited to DyLight 350, DyLight 405, DyLight 488, DyLight 550, DyLight 594, DyLight 633, DyLight 650, DyLight 680, DyLight 750 and DyLight 800 (available from Thermo Scientific (USA)). In some embodiments, the fluorophore can include but is not limited to a FluoProbes 390, FluoProbes 488, FluoProbes 532, FluoProbes 547H, FluoProbes 594, FluoProbes 647H, FluoProbes 682, FluoProbes 752 and FluoProbes 782, AMCA, DEAC (7-Diethylaminocoumarin-3-carboxylic acid); 7-Hydroxy-4-methylcoumarin-3; 7-Hydroxycoumarin-3; MCA (7-Methoxycoumarin-4-acetic acid); 7-Methoxycoumarin-3; AMF (4′-(Aminomethyl)fluorescein); 5-DTAF (5-(4,6-Dichlorotriazinyl)aminofluorescein); 6-DTAF (6-(4,6-Dichlorotriazinyl)aminofluorescein); FAM; 6-FAM (6-Carboxyfluorescein), 5(6)-FAM cadaverine; 5-FAM cadaverine; 5(6)-FAM ethylenediamme; 5-FAM ethylenediamme; 5-FITC (FITC Isomer I; fluorescein-5-isothiocyanate); 5-FITC cadaverin; Fluorescein-5-maleimide; 5-IAF (5-Iodoacetamidofluorescein); 6-JOE (6-Carboxy-4′,5′-dichloro-2′,7′-dimethoxyfluorescein); 5-CRI 10 (5-Carboxyrhodamine 110); 6-CRI 10 (6-Carboxyrhodamine 110); 5-CR6G (5-Carboxyrhodamine 6G); 6-CR6G (6-Carboxyrhodamine 6G); 5(6)-Carboxyrhodamine 6G cadaverine; 5(6)-Caroxyrhodamine 6G ethylenediamme; 5-ROX (5-Carboxy-X-rhodamine); 6-ROX (6-Carboxy-X-rhodamine); 5-TAMRA (5-Carboxytetramethylrhodamine); 6-TAMRA (6-Carboxytetramethylrhodamine); 5-TAMRA cadaverine; 6-TAMRA cadaverine; 5-TAMRA ethylenediamme; 6-TAMRA ethylenediamme; 5-TMR C6 maleimide; 6-TMR C6 maleimide; TR C2 maleimide; TR cadaverine; 5-TRITC; G isomer (Tetramethylrhodamine-5-isothiocyanate); 6-TRITC; R isomer (Tetramethylrhodamine-6-isothiocyanate); Dansyl cadaverine (5-Dimethylaminonaphthalene-1-(N-(5-aminopentyl))sulfonamide); EDANS C2 maleimide; fluorescamine; NBD; and pyrromethene and derivatives thereof.


In some embodiments, a cargo comprises an environmentally sensitive fluorescent dye or fluorophore. Examples of environmentally sensitive fluorescent dyes or fluorophores include 5,6-carboxy-diethyl rhodol (pH sensitive), merocyanine (membrane potential sensitive), and Nile red carboxylic acid (lipid sensitive).


In some embodiments, the fluorescent moiety is a peptide or protein. In some embodiments, the fluorescent moiety is Green Fluorescent Protein (GFP). In some embodiments, the fluorescent moiety is a derivative or variant of GFP, including for example: EGFP, emerald, superfolder GFP, azami green mWasabi, TagGFP, TurboGFP, AcGFP, ZsGreen, T-Sapphire, EBFP, EBFP2, Azurite, mTagBFP, ECFP, mECFP, Cerulean, m Turquoise, CyPet, AmCyan1, Midori-Ishi Cyan, TagCFP, mTFP1 (Teal), EYFP, Topaz, Venus, mCitrine, YPet, TagYFP, PhiYFP, ZsYellow1, mBanana, Kusabira Orange, Kusbira Orange2, mOrange, mOrange2, dTomato, dTomato-Tandem, TagRFP, TagRFP-Y, DsRed, DsRed2, DsRed-Express (T1), DsRed-Monomer, mTangerine, mRuby, mApple, mStrawberry, AsRed2, mRFP1, JRed, mCherry, HcRed1, mRaspberry, dKeima-Tandem, HcRed-Tandem, mPlum, AQ143, mKalamal, YFP, and Citrine.


Fluorescent moieties are detected by any suitable method. For example, a fluorescent moiety may be detected by exciting the fluorochrome with the appropriate wavelength of light and detecting the resulting fluorescence, e.g., by microscopy, visual inspection, via photographic film, by the use of electronic detectors such as charge coupled devices (CCDs), photomultipliers, etc.


In some embodiments, the fluorescent moiety is conjugated to high molecular weight molecule, such as water soluble polymers including, but not limited to, dextran, PEG, serum albumin, or poly(amidoamine) dendrimer.


Photosensitizing Agents

In some embodiments, a cargo molecule of a nerve targeting peptide conjugate comprises a photosensitizing agent. A photosensitizing agent is any agent or compound useful in light induced ablation therapy. Such agents, when exposed to a specific wavelength of light, react with molecular oxygen to produce singlet oxygen, which is highly cytotoxic. Thus, targeting molecules of the present invention comprising a photosensitizing agent may be used to focally injure nerves. In certain embodiments, a photosensitizing agent is a porphyrin, chlorin, or dye. Examples of photosensitizing agents include porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid, Lutetium texaphyrin, metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazines, metallo-naphthalocyanines, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine derivative, Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF), IR700DX, naphthalocyanine, and porphyrin derivatives.


In some embodiments, the photosensitizing agent is conjugated to a C-terminal cysteine residue of the human neuron or nerve targeting molecule via maleimide mediated conjugation. Preferably, the photosensitizing agent of the present disclosure is activated by light having a wavelength of between 400 nm to 700 nm. Still more preferably, the photosensitizing agent in the present disclosure is activated at 627 nm and 660 nm. An optimal light dose can be identified to generate maximal nerve killing with minimal injury to adjacent tissue.


Drugs

In some embodiments, a cargo molecule of a nerve targeting peptide conjugate comprises a drug. All drugs that act on a neuron or nerve (or a component thereof) are encompassed within the term “drug.” Specific examples of drug given herein, are illustrative and are not meant to limit the drugs for use with the targeting molecules disclosed herein.


In some embodiments, the drug is selected from a drug that: induces cell death (apoptotic or necrotic), inhibits cell death (apoptotic or necrotic), inhibits the transmission of a neuron or nerve signal (i.e., an electrochemical impulse), inhibits the release of a neurotransmitter, agonizes the activity of a GABA receptor, partially or fully inhibits the repolarization of a neuron, disrupts the conduction of an ion channel, or a combination thereof.


In some embodiments, the drug is an antihistamine, a GABA receptor modulator, a neurotransmitter reuptake inhibitor, a local anesthetic, an anticholinergic, a sodium channel blocker, a calcium channel blocker, a thyrotropin-releasing hormone, a y-secretase inhibitor, an AMPA receptor agonist or antagonist, an NMDA receptor agonist or antagonist, an mGlu receptor agonist or antagonist, a growth factor, an antiemetic agent, a corticosteroid; a cytotoxic agent; an antioxidant, an iron chelator, a mitochondrial modulator, a sirtuin modulator, a nitric oxide (NO) and/or nitric oxide synthase (NOS) modulator, a potassium channel agonist or antagonist, a purigenic receptor agonist or antagonist, or a combination thereof.


In some embodiments, the drug is meclizine, diphenhydramine, dimenhydrinate, loratadine, quetiapine, mepyramine, piperoxan, antazoline, carbinoxamine, doxylamine, clemastine, pheniramine, chlorphenamine, chlorpheniramine, dexchlorpheniramine, brompheniramine, triprolidine, cyclizine, chlorcyclizine, hydroxyzine, promethazine, alimemazine, trimeprazine, cyproheptadine, azatadine, ketotifen, oxatomide, meclizine hydrochloride, promethazine hydrochloride, cinnarizine, hydroxyzine pamoate, betahistine dihydrochloride, alprazolam, bromazepam, brotizolam, chlordiazepoxide, clonazepam, clorazepate, diazepam, estazolam, flunitrazepam, flurazepam, loprazolam, lorazepam, lormetazepam, idazolam, nimetazepam, nitrazepam, oxazepam, prazepam, temazepam, triazolam, clonazepam, diazepam, lorazepam, furosemide, bumetanide, ethacrynic acid, gabapentin, pregabalin, muscimol, baclofen, amitriptyline, nortriptyline, trimipramine, fluoxetine, paroxetine, sertraline, glycopyrrolate, homatropine, scopolamine, atropine, benzocaine, carticaine, cinchocaine, cyclomethycaine, lidocaine, prilocaine, propxycaine, proparacaine, tetracaine, tocainide, trimecaine, carbamazepine, oxcarbazepine, phenytein, valproic acid, sodium valproate, cinnarizine, flunarizine, nimodipine, thyrotropin-releasing hormone, amifostine (also known as WR-2721, or ETHYOL®); a carbamate compound (e.g., 2-phenyl-1,2-ethanediol monocarbomates and dicarbamates); LY450139 (hydroxylvaleryl monobenzocaprolactam); L685458 (1S-benzyl-4R[1-[1-S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarbamoyl]-2R-hydroxy-5-phenylpentyl}carbamic acid tert-butyl ester); LY411575 (N2-[(2S)-2-(3,5-difluorophenyl)-2-hydroxyethanoyl]-N1[(7S)-5-methyl-6-oxo-6,7-dihydro-5H-dibenzo[bid]azepin-7yl]-L-alaninamide); MK-0752; tarenflurbil; BMS-299897 (2-[(1R)-1-[[(4-chlorophenyl) sulfony](2,5-difluorophenyl)amino]ethyl]-5-fluorobenzenepropanoic acid; CNQX (6-cyano-7-nitroquinoxaline-2,3-dione); NBQX (2,3-dihydroxy-6-nitro-7-sulfamoylbenzo[f]quinoxaline-2,3-dione); DNQX (6,7-dinitroquinoxaline-2,3-dione); kynurenic acid; 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo-[fjquinoxaline; 1-aminoadamantane; dextromethorphan; dextrorphan; ibogaine; ketamine; nitrous oxide; phencyclidine; riluzole; tiletamine; memantine; dizocilpine; aptiganel; remacimide; 7-chlorokynurenate; DCKA (5,7-dichlorokynurenic acid); kynurenic acid; 1-aminocyclopropanecarboxylic acid (ACPC); AP7 (2-amino-7-phosphonoheptanoic acid); APV (R-2-amino-5-phosphonopentanoate); CPPene (3-[(R)-2-carboxypiperazin-4-yl]-prop-2-enyl-1-phosphonic acid); (+)-(1S,2S)-1-(4-hydroxy-phenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-pro-panol; (1S,2S)-1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-phenylpiperi-dino)-1-propanol; (3R,4S)-3-(4-(4-fluorophenyl)-4-hydroxypiperidin-1-yl-)-chroman-4;7-diol; (1R*,2R*)-1-(4-hydroxy-3-memylphenyl)-2-(4-(4-fluoro-phenyl)-4-hydroxypiperidin-1-yl)-propan-1-ol-mesylate); LY389795 ((−)-2-thia-4-aminobicyclo-hexane-4,6-dicarboxylate); LY379268 ((−)-2-oxa-4-aminobicyclo-hexane-4,6-dicarboxylate); LY354740 ((+)-2-aminobicyclo-hexane-2,6dicarboxylate); DCG-IV ((2S,2′R,3′R)-2-(2′,31-dicarboxycyclopropyl)glycine); 2R,4R-APDC (2R,4R-4-aminopyrrolidine-2,4-dicarboxylate); (S)-3C4HPG ((S)-3-carboxy-4-hydroxyphenylglycine); (S)-4C3HPG ((S)-4-carboxy-3-hydroxyphenylglycine); L-CCG-I ((2S,1′S,2′S)-2-(carboxycyclopropyl)glycine); ACPT-I ((1S,3R,4S)-1-aminocyclopentane-1,3,4-tricarboxylic acid); L-AP4 (L-(+)-2-Amino-4-phosphonobutyric acid); (S)-3,4-DCPG ((S)-3,4-dicarboxyphenylglycine); (RS)-3,4-DCPG ((RS)-3,4-dicarboxyphenylglycine); (RS)-4-phosphonophenylglycine ((RS)PPG); AMN082 (,N′-bis(diphenylmethyl)-1,2-ethanediamine dihydrochloride); DCG-IV ((2S,2′R,3′R)-2-(2′,3′-dicarboxycyclopropyl)glycine); AMN082; brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived neurotrophic factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-like growth factor (IGF); an aminoglycoside antibiotic (e.g., gentamicin and amikacin); a macrolide antibiotic (e.g, erythromycin); a glycopeptide antibiotic (e.g. vancomycin); salicylic acid; nicotine; Eburnamenine-14-carboxylic acid ethyl ester; sipatrigine (2-(4-Methylpiperazin-1-yl)-5-(2,3,5-trichlorophenyl)-pyrimidin-4-amine); amiloride (3,5-diamino-N-(aminoiminomethyl)-6-chloropyrazinecarbox amide hydrochloride); carbamazepine (5H-dibenzo[b,f]azepine-5-carboxamide); TTX (octahydro-12-(hydroxymethyl)-2-imino-5,9:7,10a-dimethano-10aH-[1,3]dioxocino[6,5-d]pyrimidine-4,7,10,11,12-pentol); RS100642 (1-(2,6-dimethyl-phenoxy)-2-ethylaminopropane hydrochloride); mexiletine ((1-(2,6-dimethylphenoxy)-2-aminopropane hydrochloride)); QX-314 (N-(2,6-Dimethylphenylcarbamoylmethyl)triethylammonium bromide); phenytoin (5,5-diphenylimidazolidine-2,4-dione); lamotrigine (6-(2,3-dichlorophenyl)-1,2,4-triazine-3,5-diamine); 4030W92 (2,4-diamino-5-(2,3-dichlorophenyl)-6-fluoromethylpyrimidine); BW1003C87 (5-(2,3,5-trichlorophenyl)pyrimidine-2,4-1.1 ethanesulphonate); QX-222 (2-[(2,6-dimethylphenyl)amino]-N,N,N-trimethyl-2-oxoetha niminium chloride); ambroxol (trans-4-[[(2-Amino-3,5-dibromophenyl)methyl]amino]cyclo hexanol hydrochloride); R56865 (N-[1-(4-(4-fluorophenoxy)butyl]-4-piperidinyl-N-methyl-2-benzo-thiazolamine); lubeluzole; ajmaline ((17R,21alpha)-ajmalan-17,21-diol); procainamide (4-amno-N-(2-diethylaminoethyljbenzamide hydrochloride); flecainide; riluzoleor; triamicinolone actenoide; Dexamethasone; promethazine; prochlorperazine; trimethobenzamide; triethylperazine; dolasetron; granisetron; ondansetron; tropisetron; and palonosetron; droperidol; meclizine; perphenazine; thiethyl perazine; domperidone; properidol; haloperidol; chlorpromazine; promethazine; prochlorperazine; metoclopramide; dronabinol; nabilone; sativex; scopolamine; dexamethasone; trimethobenzamine; emetrol; propofol; muscimol; acridine carboxamide; actinomycin; 17-N-allylamino-17-demethoxygeldanamycin; amcinonide; amsacrine; aminopterin; anthracycline; antineoplastic; antineoplaston; 5-azacytidine; azathioprine; BL22; beclometasone; bendamustine; betamethasone; biricodar; bleomycin; bortezomib; bryostatin; budesonide; busulfan; calyculin; camptothecin; capecitabine; carboplatin; chlorambucil; cisplatin; cladribine; clofarabine; cortisone acetate; cytarabine; dacarbazine; dasatinib; daunorubicin; decitabine; desonide; dexamethasone; dichloroacetic acid; discodermolide; docetaxel; doxorubicin; epirubicin; epothilone; eribulin; estramustine; etoposide; exatecan; exisulind; ferruginol; floxuridine; fludarabine; fluocinonide; fluocortolone; fluorouracil; fosfestrol; fotemustine; gemcitabine; halometasone; halcinonide; hydrocortisone; hydrocortisone acetate; hydroxyurea; IT-101; idarubicin; ifosfamide; imiquimod; irinotecan; irofulven; ixabepilone; laniquidar; lapatinib; lenalidomide; lomustine; lurtotecan; mafosfamide; masoprocol; mechlorethamine; melphalan; mercaptopurine; methylprednisone; mitomycin; mitotane; mitoxantrone; mometasone; nelarabine; nilotinib; oblimersen; oxaliplatin; PAC-I; methotrexate (RHEUMATREX®, Amethopterin); cyclophosphamide (CYTOXAN®) thalidomide (THALID OMID®); paclitaxel; pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; prednisolone; prednisone; procarbazine; proteasome inhibitors (e.g., bortezomib); raltitrexed; rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; thioTEPA; tioguanine; topotecan; trabectedin; triamcinolone acetonide; tretinoin; triplatin tetranitrate; tris(2-chloroethyl)amme; troxacitabine; uracil mustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; N-acetylcysteine; vitamin E; vitamin C; vitamin A; lutein; selenium glutathione; melatonin; a polyphenol; a carotenoid; coenzyme Q-IO; Ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one (also called PZ 51 or DR3305); L-methionine; azulenyl nitrones; L-(+)-Ergothioneine; CAPE (caffeic acid phenethyl ester); dimethylthiourea; dimethylsulfoxide; disufenton sodium; pentoxifylline; MCI-186; Ambroxol; U-83836E; MitoQ (mitoquinone mesylate); Idebenone (2-(10-hydroxydecyl)-5,6-dimethoxy-3-methyl-cyclohexa-2,5-diene-1,4-dione); desferrioxamine; hydroxybenzyl ethylene diamine; fullerenol-1, pyrrolidine dithiocarbamate; acetylcarnitine; lipoic acid; a stilbene; a chalcone; a flavone; an isoflavone; a flavanones; an anthocyanidin; a catechin; isonicotinamide; dipyridamole; ZM 336372; camptothecin; coumestrol; nordihydroguaiaretic acid; esculetin; SRT-1720; SRT-1460; SRT-2183; aminoguanidine; 1-Amino-2-hydroxyguanidine p-toluensulfate; GED; bromocriptine mesylate; dexamethasone; SDMA; ADMA; L-NMMA; L-NMEA; D-MMA; L-NIL; L-NNA; L-NPA; L-NAME; L-VNIO; diphenyleneiodonium chloride; 2-ethyl-2-thiopseudourea; haloperidol; L-NIO; MEG; SMT; SMTC; 7-Ni; nNOS inhibitor; 1,3-PBITU; L-thiocitrulline; TRIM; MTR-105; BBS-I; BBS-2; ONO-1714; GW273629; GW 274150; PPA250; AR-R17477; AR-R18512; spiroquinazolone; 1400W; S-NC; NTG; SNP; thapsigargin; VEGF; bradykinin; ATP; sphingosine-1-phosphate; estrogen; angiopoietin; acetylcholine; SIN-I; GEA 3162; GEA; GEA 5024; GEA 5538; SNAP; molsidomine; CNO-4; CNO-5; DEA/NO; IPA/NO; SPER/NO; SULFI/NO; OXI/NO; DETA/NO; nicorandil; minoxidil, levcromakalim; lemakalim; cromakalim; L-735,334; retigabine; flupirtine; BMS-204352; DMP-543; linopirdine; XE991; 4-AP; 3,4-DAP; E-4031; DIDS; Way 123,398; CGS-12066 A; dofetilide; sotalol; apamin; amiodarone; azimilide; bretylium; clofilium; tedisamil; ibutilide; sematilide; nifekalant; tamulustoxin; ATP; ADP; UTP; UDP; UDP-glucose; adenosine; 2-MESATP; 2-MESADP; ABMEATP; DATPAS; ATPrS; BZ-ATP; MRS2703; DENUFOSOL TETRASODIUM; MRS2365; MRS 2690; PSB 0474; A-317491; RO-3 (Roche); SURAMIN; PPADS; PPNDS; DIDS; pyridoxal-5-phosphate; 5-(3-bromophenyl)-1,3-dihydro-2H-benzofuro-[3,2-e]-1,4-diazepin-2-one; cibacron blue; basilen blue; ivermectin; A-438079; A-740003; NF023; NF449; NFI 10; NF157; MRS 2179; NF279; MRS 2211; MRS 2279; MRS 2500 tetrasodium salt; TNP-ATP; tetramethylpyrazine; Ip51; jQy-carboxymethylene ATP; βγ-chlorophosphomethylene ATP; KN-62; spinorphin; minocycline; SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl) 1H-imidazole); PD 169316 (4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole); SB 202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole); RWJ 67657 (4-[4-(4-fluorophenyl)-1-(3-phenylpropyl)-5-(4-pyridinyl)-1H-imidazol-2-yl]-3-butyn-1-ol); SB 220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole); D-JNKI-I ((D)-hJIPi75-i57-DPro-DPro-(D)-HIV-T AT57-48); AM-111 (Auris); SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one); JNK Inhibitor I ((L)-HIV-TAT48-57-PP-JBD20); JNK Inhibitor III ((L)-HIV-TAT47-57-gaba-c-Jun833-57); AS601245 (1,3-benzothiazol-2-yl (2-[[2-(3-pyridinyl)ethyl]amino]-4 pyrimidinyl) acetonitrile); JNK Inhibitor VI (H2N-RPKRPTTLNLF-NH2) (SEQ ID NO:564); JNK Inhibitor VIII (N-(4-Amino-5-cyano-6-ethoxypyridin-2-yl)-2-(2,5-dimethoxyphenyl)acetamide); JNK Inhibitor IX (N-(3-Cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl)-1-naphthamide); dicumarol (3,3′-Methylenebis(4-hydroxycoumarin)); SC-236 (4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzene-sulfonamide); CEP-1347 (Cephalon); CEP-11004 (Cephalon); an artificial protein comprising at least a portion of a Bcl-2 polypeptide; a recombinant FNK; V5 (also known as Bax inhibitor peptide V5); Bax channel blocker ((+)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol); Bax inhibiting peptide P5 (also known as Bax inhibitor peptide P5); Kp7-6; FAIM(S) (Fas apoptosis inhibitory molecule-short); FAIM(L) (Fas apoptosis inhibitory molecule-long); Fas:Fc; FAP-I; NOK2; F2051; F1926; F2928; ZB4; Fas M3 mAb; EGF; 740 Y-P; SC 3036 (KKHTDDGYMPMSPGVA) (SEQ ID NO:565); PI 3-kinase Activator (Santa Cruz Biotechnology, Inc.); Pam3Cys ((S)-(2,3-bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys4-OH, trihydrochloride); Actl (NF-kB activator 1); an anti-DcB antibody; Acetyl-11-keto-b-Boswellic Acid; Andrographolide; Caffeic Acid Phenethyl Ester (CAPE); Gliotoxin; Isohelenin; NEMO-Binding Domain Binding Peptide (DRQIKIWFQNRRMKWKKTALDWSWLQTE) (SEQ ID NO:566); NF-kB Activation Inhibitor (6-Amino-4-(4-phenoxyphenylethylamino)quinazoline); NF-kB Activation Inhibitor II (4-Methyl-Nl-(3-phenylpropyl)benzene-1,2-diamine); NF-kB Activation Inhibitor III (3-Chloro-4-nitro-N-(5-nitro-2-thiazolyl)-benzamide); NF-kB Activation Inhibitor IV ((E)-2-Fluoro-4′-methoxystilbene); NF-kB Activation Inhibitor V (5-Hydroxy-(2,6-diisopropylphenyl)-1H-isoindole-1,3-dione); NF-kB SN50 (AAV ALLP A VLLALL AP VQRKRQKLMP) (SEQ ID NO:567); Oridonin; Parthenolide; PPM-18 (2-Benzoylamino-1,4-naphthoquinone); Rol06-9920; Sulfasalazine; TIRAP Inhibitor Peptide (RQIKiWFNRRMKWKKLQLRD AAPGGAIVS) (SEQ ID NO:568); Withaferin A; Wogonin; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile); BAY 11-7085 ((E)3-[(4-t-Butylphenyl)sulfonyl]-2-propenenitrile); (E)-Capsaicin; Aurothiomalate (ATM or AuTM); Evodiamine; Hypoestoxide; IKK Inhibitor III (BMS-345541); IKK Inhibitor VII; IKK Inhibitor X; IKK Inhibitor II; IKK-2 Inhibitor IV; IKK-2 Inhibitor V; IKK-2 Inhibitor VI; IKK-2 Inhibitor (SC-514); IkB Kinase Inhibitor Peptide; IKK-3 Inhibitor IX; ARRY-797 (Array BioPharma); SB-220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole); SB-239063 (trans-4-[4-(4-Fluorophenyl)-5-(2-methoxy-4-pyrimidinyl)-1H-imidazol-1-ylJcyclohexanol); SB-202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole); JX-401 (-[2-Methoxy-4-(methylthio)benzoyl]-4-(phenylmethyl)piperidine); PD-169316 (4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole); SKF-86002 (6-(4-Fluorophenyl)-2,3-dihydro-5-(4-pyridinyl)imidazo[2,1-b]thiazole dihydrochloride); SB-200646 (N-(1-Methyl-1H-indol-5-yl)-N′-3-pyridinylurea); CMPD-I (2′-Fluoro-N-(4-hydroxyphenyl)-[1,1′-biphenyl]-4-butanamide); EO-1428 ((2-Methylphenyl)-[4-[(2-amino-4-bromophenyl)amino]-2-chlorophenyl]methanone); SB-253080 (4-[5-(4-Fluorophenyl)-2-[4-(methylsulfonyl)phenyl]-1H-imidazol-4-yl]pyridine); SD-169 (1H-Indole-5-carboxamide); SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfinyl phenyl)-5-(4-pyridyl) 1H-imidazole); TZP-101 (Tranzyme Pharma); TZP-102 (Tranzyme Pharma); GHRP-6 (growth hormone-releasing peptide-6); GHRP-2 (growth hormone-releasing peptide-2); EX-1314 (Elixir Pharmaceuticals); MK-677 (Merck); L-692,429 (Butanamide, 3-amino-3-methyl-N-(2,3,4,5-tetrahydro-2-oxo-1-((2′-(1H-tetrazol-5-yl)(1,1′-biphenyl)-4-yl)methyl)-1H-1-benzazepin-3-yl)-, (R)-); EP1572 (Aib-DTrp-DgTrp-CHO); diltiazem; metabolites of diltiazem; BRE (Brain and Reproductive organ-Expressed protein); verapamil; nimodipine; diltiazem; omega-conotoxin; GVIA; amlodipine; felodipine; lacidipine; mibefradil; NPPB (5-Nitro-2-(3-phenylpropylamino)benzoic Acid); flunarizine; erythropoietin; pipeline; hemin; brazilin; z-VAD-FMK (Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone); z-LEHD-FMK (benzyloxycarbonyl-Leu-Glu(OMe)-His-Asp(OMe)-fluoromethylketone) (SEQ ID NO:569); B-D-FMK (boc-aspartyl(Ome)-fluoromethylketone); Ac-LEHD-CHO (N-acetyl-Leu-Glu-His-Asp-CHO) (SEQ ID NO:569); Ac-IETD-CHO (N-acetyl-Ile-Glu-Thr-Asp-CHO) (SEQ ID NO:570); z-IETD-FMK (benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)-fluoromethylketone) (SEQ ID NO:570); FAM-LEHD-FMK (benzyloxycarbonyl Leu-Glu-His-Asp-fluoromethyl ketone) (SEQ ID NO:569); FAM-LETD-FMK (benzyloxycarbonyl Leu-Glu-Thr-Asp-fluoromethyl ketone) (SEQ ID NO:571); Q-VD-OPH (Quinoline-Val-ASp-CH2-O-Ph); XIAP; cIAP-1; cIAP-2; ML-IAP; ILP-2; NAIP; Survivin; Bruce; IAPL-3; fortilin; leupeptine; PD-150606 (3-(4-Iodophenyl)-2-mercapto-(Z)-2-propenoic acid); MDL-28170 (Z-Val-Phe-CHO); calpeptin; acetyl-calpastatin; MG 132 (N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(1S)-1-formyl-3-methylbutyl]-L-leucinamide); MYODUR; BN 82270 (Ipsen); BN 2204 (Ipsen); AHLi-11 (Quark Pharmaceuticals), an mdm2 protein, pifithrin-α (1-(4-Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl)ethanone); trans-stilbene; cis-stilbene; resveratrol; piceatannol; rhapontin; deoxyrhapontin; butein; chalcon; isoliquirtigen; butein; 4,2′,4′-trihydroxychalcone; 3,4,2′,4′,6′-pentahydroxychalcone; flavone; morin; fisetin; luteolin; quercetin; kaempferol; apigenin; gossypetin; myricetin; 6-hydroxyapigenin; 5-hydroxyflavone; 5,7,3′,4′,5′-pentahydroxyflavone; 3,7,3′,4′,5′-pentahydroxyflavone; 3,6,3′,4′-tetrahydroxyflavone; 7,3′,4′,5′-tetrahydroxyflavone; 3,6,2′,4′-tetrahydroxyflavone; 7,4′-dihydroxyflavone; 7,8,3′,4′-tetrahydroxy flavone; 3,6,2′,3′-tetrahydroxyflavone; 4′-hydroxyflavone; 5-hydroxyflavone; 5,4′-dihydroxyflavone; 5,7-dihydroxyflavone; daidzein; genistein; naringenin; flavanone; 3,5,7,3′,4′-pentahydroxyflavanone; pelargonidin chloride; cyanidin chloride; delphinidin chloride; (−)-epicatechin (Hydroxy Sites: 3,5,7,3′,4{circumflex over ( )}; (−)-catechin (Hydroxy Sites: 3,5,7,3′,40; (−)-gallocatechin (Hydroxy Sites: 3,5,7,3′,4′,5O (+)-catechin (Hydroxy Sites: 3,5,7,3′,4{circumflex over ( )}; (+)-epicatechin (Hydroxy Sites: 3,5,7,3′,41J; Hinokitiol (b-Thujaplicin; 2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien-1-one); L-(+)-Ergothioneine ((S)-a-Carboxy-2,3-dihydro-N,N,N-trimethyl-2-thioxo-1H-imidazole4-ethanaminium inner salt); Caffeic Acid Phenyl Ester; MCI-186 (3-Methyl-1-phenyl-2-pyrazolin-5-one); HBED (N,N′-Di-(2-hydroxybenzy{circumflex over ( )}ethylenediamine-HN′-diacetic acid<<H2O); Ambroxol (trans-4-(2-Amino-3,5-dibromobenzylamino)cyclohexane-HCl; and U-83836E ((−)-2-((4-(2,6-di-1-Pyrrolidinyl-4-pyrimidinyl)-1-piperzainyl)methyl)-3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol*2HCl); /5-1-5-methyl-nicotinamide-2′-deoxyribose; /S-D-I′-5-methyl-nico-tinamide-2′-deoxyribofuranoside; /3-1′-4,5-dimethyl-nicotinamide-2′-de-oxyribose; /3-D-1′-4,5-dimethyl-nicotinamide-2′-deoxyribofuranoside; 1-Naphthyl PP 1 (1-(1,1-Dimethyl ethyl)-3-(1-naphthalenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); Lavendustin A (5-[[(2,5-Dihydroxyphenyl)methyl][(2-hydroxyphenyl)methyl]amino]-2-hydroxybenzoic acid); MNS (3,4-Methylenedioxy-b-nitrostyrene); PP 1 (1-(1,1-Dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); PP2 (3-(4-chlorophenyl) 1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); KX1-004 (Kinex); KX1-005 (Kinex); KX1-136 (Kinex); KX1-174 (Kinex); KX1-141 (Kinex); KX2-328 (Kinex); KX1-306 (Kinex); KX1-329 (Kinex); KX2-391 (Kinex); KX2-377 (Kinex); ZD4190 (Astra Zeneca; N-(4-bromo-2-fluorophenyl)-6-methoxy-7-(2-(1H-1,2,3-triazol-1-yl)ethoxy)quinazolin-4-amine); AP22408 (Ariad Pharmaceuticals); AP23236 (Ariad Pharmaceuticals); AP23451 (Ariad Pharmaceuticals); AP23464 (Ariad Pharmaceuticals); AZD0530 (Astra Zeneca); AZM475271 (M475271; Astra Zeneca); Dasatinib (N-(2-chloro-6-methylphneyl)-2-(6-(4-(2-hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino) thiazole-5-carboxamide); GN963 (trans-4-(6,7-dimethoxyquinoxalin-2ylamino)cyclohexanol sulfate); Bosutinib (4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methyl-1-piperazinyl)propoxy)-3-quinolinecarbonitrile); or combinations thereof.


Radiolabels

In some embodiments, a cargo molecule comprises a radiolabel. As used herein, “radiolabel” refers to a moiety comprising a radioactive isotope of at least one element. In some embodiments, a radiolabel is one used in positron emission tomography (PET). In some embodiments, a radiolabel is one used in single-photon emission computed tomography (SPECT). In some embodiments, radioisotopes comprise 99mTc, 111In, 64Cu, 67Ga, 186Re, 188Re, 153Sm, 177Lu, 67Cu, 123I, 124I, 125I, 11C, 3N, 15O, 18F, 186Re, 188Re, 153Sm, 166Ho, 177Lu, 149Pm, 90Y, 213Bi, 103Pd, 109Pd, 159Gd, 140La, 198Au, 199Au, 169Yb, 175Yb, 165Dy, 166Dy, 67Cu, 105Rh, 111Ag, 89Zr, 225Ac, and 192Ir.


Neurotropic Factors

In some embodiments, a cargo molecule comprises a factor having neurotrophic properties (e.g., neurotrophic proteins such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), glial cell line-derived neurotrophic factor (GDNF), ciliary neurotrophic factor (CNTF) as well as non-protein small molecules with neurotrophic properties).


III. Linkers

In some embodiments, a cargo molecule (e.g., a fluorescent moiety, photosensitizing agent, or drug) is directly attached to the peptides disclosed herein, e.g. at the end of the targeting peptide. Alternatively, in some embodiments, a cargo molecule (e.g., a fluorescent moiety, photosensitizing agent, or drug) is indirectly attached to a peptide disclosed herein, e.g., via a linker.


In some embodiments of the nerve targeting peptide conjugates (with or without a linker) described herein, the peptide comprises or consists or the amino acid sequence set forth in any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS: 1-30. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:31-36. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:37-40. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:41-240. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:241-349. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:350-449. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:450-557.


In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of Formulas (I)-(XLIV) or Subformulas (Ia)-(XLIVa).


As used herein, a “linker” is any molecule capable of binding (e.g., covalently) to a targeting molecule disclosed herein. Linkers include, but are not limited to, straight or branched-chain carbon linkers, heterocyclic carbon linkers, amino acid linkers (e.g., D- or L-amino acid), lipophilic residues, peptide linkers, peptide nucleic acid linkers, hydrazone linkers, SPDB disulfide, sulfo-SPDB, maleimidomethyl cyclohexane-1-carboxylate (MCC), aminohexanoic acid linkers, and polyether linkers (e.g., PEG). For example, poly(ethylene glycol) linkers are available from Quanta Biodesign, Powell, OH. These linkers optionally have amide linkages, sulfhydryl linkages, or hetero functional linkages.


In some embodiments, the linker binds to a targeting molecule disclosed herein by a covalent linkage. A linker may connect a cargo molecule to the peptide by forming a covalent linkage to the cargo molecule at one location and a covalent linkage to the peptide at another location. The covalent linkages can be formed by reaction between functional groups on the linker and functional groups on the peptide and on the cargo molecule. In some embodiments, the covalent linkage comprises an ether bond, thioether bond, amine bond, amide bond, carbon-carbon bond, carbon-nitrogen bond, carbon-oxygen bond, or carbon-sulfur bond.


In some embodiments, the linker is flexible. In some embodiments, the linker is rigid. In some embodiments, the linker has segment(s) of flexibility and segment(s) of rigidity.


In some embodiments, the linker comprises a linear structure. In some embodiments, the linker comprises a non-linear structure. In some embodiments, the linker comprises a branched structure. In some embodiments, the linker comprises a cyclic structure.


In some embodiments, the linker is an alkyl. In some embodiments, the linker is a heteroalkyl.


In some embodiments, the linker is an alkylene. In some embodiments, the linker is an alkenylene. In some embodiments, the linker is an alkynylene. In some embodiments, the linker is a heteroalkylene.


An “alkyl” group refers to an aliphatic hydrocarbon group. The alkyl moiety may be a saturated alkyl or an unsaturated alkyl. Depending on the structure, an alkyl group can be a monoradical or a diradical (i.e., an alkylene group).


The “alkyl” moiety may have 1 to 10 carbon atoms (whenever it appears herein, a numerical range such as “1 to 10” refers to each integer in the given range; e.g., “1 to 10 carbon atoms” means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms, although the present definition also covers the occurrence of the term “alkyl” where no numerical range is designated). The alkyl group could also be a “lower alkyl” having 1 to 6 carbon atoms. The alkyl group of the compounds described herein may be designated as “Ci-C4 alkyl” or similar designations. By way of example only, “C1-C4 alkyl” indicates that there are one to four carbon atoms in the alkyl chain, i.e., the alkyl chain is selected from methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl. Typical alkyl groups include, but are in no way limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, ethenyl, propenyl, butenyl, and the like.


In some embodiments, the linker comprises a ring structure (e.g., an aryl). As used herein, the term “ring” refers to any covalently closed structure. Rings include, for example, carbocycles (e.g., aryls and cycloalkyls), heterocycles (e.g., heteroaryls and non-aromatic heterocycles), aromatics (e.g. aryls and heteroaryls), and non-aromatics (e.g., cycloalkyls and non-aromatic heterocycles). Rings can be optionally substituted. Rings can be monocyclic or polycyclic.


As used herein, the term “aryl” refers to an aromatic ring wherein each of the atoms forming the ring is a carbon atom. Aryl rings can be formed by five, six, seven, eight, nine, or more than nine carbon atoms. Aryl groups can be optionally substituted. Examples of aryl groups include, but are not limited to phenyl, naphthalenyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl. Depending on the structure, an aryl group can be a monoradical or a diradical (i.e., an arylene group).


The term “cycloalkyl” refers to a monocyclic or polycyclic non-aromatic radical, wherein each of the atoms forming the ring (i.e. skeletal atoms) is a carbon atom. Cycloalkyls may be saturated, or partially unsaturated. Cycloalkyl groups include groups having from 3 to 10 ring atoms. Cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.


In some embodiments, the ring is a cycloalkane. In some embodiments, the ring is a cycloalkene.


In some embodiments, the ring is an aromatic ring. The term “aromatic” refers to a planar ring having a delocalized π-electron system containing 4n+2 π electrons, where n is an integer. Aromatic rings can be formed from five, six, seven, eight, nine, or more than nine atoms. Aromatics can be optionally substituted. The term “aromatic” includes both carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or “heteroaryl” or “heteroaromatic”) groups (e.g., pyridine). The term includes monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups.


In some embodiments, the ring is a heterocycle. The term “heterocycle” refers to heteroaromatic and heteroalicyclic groups containing one to four heteroatoms each selected from O, S and N, wherein each heterocyclic group has from 4 to 10 atoms in its ring system, and with the proviso that the ring of said group does not contain two adjacent O or S atoms. Non-aromatic heterocyclic groups include groups having only 3 atoms in their ring system, but aromatic heterocyclic groups must have at least 5 atoms in their ring system. The heterocyclic groups include benzo-fused ring systems. An example of a 3-membered heterocyclic group is aziridinyl. An example of a 4-membered heterocyclic group is azetidinyl (derived from azetidine). An example of a 5-membered heterocyclic group is thiazolyl. An example of a 6-membered heterocyclic group is pyridyl, and an example of a 10-membered heterocyclic group is quinolinyl. Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, 3H-indolyl and quinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and faropyridinyl. The foregoing groups, may be C-attached or N-attached where such is possible. For instance, a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group derived from imidazole may be imidazol-1-yl or imidazol-3-yl (both N-attached) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl (all C-attached). The heterocyclic groups include benzo-fused ring systems and ring systems substituted with one or two oxo (═O) moieties such as pyrrolidin-2-one. Depending on the structure, a heterocycle group can be a monoradical or a diradical (i.e., a heterocyclene group).


In some embodiments, the ring is fused. The term “fused” refers to structures in which two or more rings share one or more bonds, hi some embodiments, the ring is a dimer. In some embodiments, the ring is a trimer. In some embodiments, the ring is a substituted.


The term “carbocyclic” or “carbocycle” refers to a ring wherein each of the atoms forming the ring is a carbon atom. Carbocycle includes aryl and cycloalkyl. The term thus distinguishes carbocycle from heterocycle (“heterocyclic”) in which the ring backbone contains at least one atom which is different from carbon (i.e., a heteroatom). Heterocycle includes heteroaryl and heterocycloalkyl. Carbocycles and heterocycles can be optionally substituted.


In some embodiments, the linker is substituted. The term “optionally substituted” or “substituted” means that the referenced group may be substituted with one or more additional group(s) individually and independently selected from Ci-Cealkyl, C3-Cgcycloalkyl, aryl, heteroaryl, C2-C6heteroalicyclic, hydroxy, Ci-C6alkoxy, aryloxy, Ci-C6alkylthio, arylthio, Ci-C6alkylsulfoxide, arylsulfoxide, Ci-C6alkylsulfone, arylsulfone, cyano, halo, C2-C8acyl, C2-C8acyloxy, nitro, Ci-C6haloalkyl, Ci-C6fluoroalkyl, and amino, including Ci-C6alkylamino, and the protected derivatives thereof. By way of example, an optional substituents may be LSRS, wherein each Ls is independently selected from a bond, —O—, —C(═O)—, —S—, —S(═O)—, —S(O)2-, —NH—, —NHC(O)—, —C(O)NH—, S(O)2NH—, —NHS(O)2-, —OC(O)NH—, —NHC(O)O—, -(CpC6alkyl)-, or —(C2-C6alkenyl)-; and each Rs is independently selected from H, (Ci-C4alkyl), (C3-C8cycloalkyl), heteroaryl, aryl, and Ci-C6heteroalkyl. Optionally substituted non-aromatic groups may be substituted with one or more oxo (═O). The protecting groups that may form the protective derivatives of the above substituents are known to those of skill in the art.


In some embodiments, a bifunctional linker having one functional group reactive with a group on one molecule (e.g., a targeting molecule), and another group reactive on the other molecule (e.g., a fluorescent moiety or a drug), is used to form the desired conjugate. Alternatively, in some embodiments, derivatization is performed to provide functional groups. Thus, for example, procedures for the generation of free sulfhydryl groups on peptides are also known (See U.S. Pat. No. 4,659,839). A linker may alternatively comprise a heterobifunctional crosslinker comprising two or more different reactive groups that form a heterocyclic ring that can interact with a targeting molecule. For example, a heterobifunctional crosslinker such as cysteine may comprise an amine reactive group and a thiol-reactive group can interact with an aldehyde on a derivatized targeting molecule. Additional combinations of reactive groups suitable for heterobifunctional crosslinkers include, for example, amine- and sulfhydryl reactive groups; carbonyl and sulfhydryl reactive groups; amine and photoreactive groups; sulfhydryl and photoreactive groups; carbonyl and photoreactive groups; carboxylate and photoreactive groups; and arginine and photoreactive groups. Examples of a heterobifunctional crosslinker include N-Succinimidyl 4-(2-pyridyldithio)butanoate (SPDB) and maleimidomethyl cyclohexane-1-carboxylate (MCC).


In some embodiments, a linker is cleavable. In some embodiments, the linker is non-cleavable. A linker can be chemically stable to extracellular environments, for example, chemically stable in the blood stream, and/or may include linkages that are not stable. A linker can include linkages that are designed to cleave and/or immolate or otherwise breakdown specifically or non-specifically inside cells. A cleavable linker can be sensitive to enzymes at a specific site, such as by extracellular proteases.


A cleavable linker can include a valine-citrulline peptide, a valine-alanine peptide, a phenylalanine-lysine or other peptide, such as a peptide that forms a protease recognition and cleavage site. Such a peptide-containing linker can contain a pentafluorophenyl group. A peptide-containing linker can include a succimide or a maleimide group. A peptide-containing linker can include a para aminobenzoic acid (PABA) group. A peptide-containing linker can include an aminobenzyloxycarbonyl (PABC) group. A peptide-containing linker can include a PABA or PABC group and a pentafluorophenyl group. A peptide-containing linker can include a PABA or PABC group and a succinimide group. A peptide-containing linker can include a PABA or PABC group and a maleimide group.


A non-cleavable linker is generally protease-insensitive and insensitive to intracellular processes. A non-cleavable linker can include a maleimide group. A non-cleavable linker can include a succinimide group. A non-cleavable linker can be maleimido-alkyl-C(O)— linker. A non-cleavable linker can be maleimidocaproyl linker. A maleimidocaproyl linker can be N-maleimidomethylcyclohexane-1-carboxylate. A maleimidocaproyl linker can include a succinimide group. A maleimidocaproyl linker can include pentafluorophenyl group.


In some embodiments, a peptide linker consisting of one or more amino acids is used to join the targeting molecule and a fluorescent moiety or drug. Generally, the peptide linker will have no specific biological activity other than to join the molecules or to preserve some minimum distance or other spatial relationship between them. However, the constituent amino acids of the linker may be selected to influence some property of the molecule such as the folding, net charge, or hydrophobicity. In some embodiments the peptide linker is relatively short, typically less than about 10 amino acids, preferably less than about 8 amino acids and more preferably less than 5 amino acids. Non-limiting illustrative examples include glycine and glycine-serine linkers which can be added to the C-terminus of a peptide. In some embodiments, a peptide linker comprises a cysteine residue or an unnatural amino acid residue (e.g., selenocysteine (Sec), p-acetophenylalanine (pAcF), p-azidomethyl-L-phenylalanine (pAMF), and azido-lysine (AzK)) for site specific conjugation. In some embodiments, a peptide linker is a glycine-glycine-glycine-cysteine (GGGC) (SEQ ID NO:572) linker, a glycine-glycine-cysteine (GGC) linker, a glycine-glycine (GG) linker, or a cysteine (C) linker. In some embodiments, the GGGC (SEQ ID NO:572), GGC, GG, or C linker is added to the N-terminus or C-terminus of a peptide.


IV. Further Modifications

In some embodiments, the peptides of the present disclosure are optionally conjugated to high molecular weight molecules that increase the multivalency and avidity of labeling. In some embodiments, the high molecular weight molecules are water-soluble polymers. Examples of suitable water-soluble polymers include, but are not limited to, peptides, saccharides, poly(vinyls), poly(ethers), poly(amines), poly(carboxylic acids) and the like. In some embodiments, the water-soluble polymers is dextran, polyethylene glycol (PEG), polyoxyalkylene, polysialic acid, starch, or hydroxyethyl starch. Any suitable method is used to conjugate peptides to water-soluble polymers (see, Hermanson G., Bioconjugate Techniques 2nd Ed., Academic Press, Inc. 2008). In some embodiments, the peptide-high molecular weight molecule conjugate further comprises a cargo molecule.


In some embodiments, one or more additional amino acid residues can be added to the N-terminus, the C-terminus, or both N-terminus and C-terminus of the peptides disclosed herein, thus producing a longer peptide. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more amino acid residues are added to the peptides disclosed herein. In some embodiments, about 1-15, 1-10, 1-5, or 1-3 amino acids are added to the peptides disclosed herein. In some embodiments, additional amino acids that are added to the peptides of the present disclosure may act as a spacer to provide a variable distance between the peptide and the cargo molecule.


In some embodiments, the nerve targeting peptide conjugates of the present invention are modified to increase solubility. Peptide modifications that increase solubility include addition of hydrophilic amino acid(s), a PEG moiety, or both. In some embodiments, a PEG moiety is 8-Amino-3,6-dioxaoctanoic acid (AEEA); 12-amino-4,7,10-trioxadodecanoic acid; or 15-amino-4,7,10,13-tetraoxapenta-decanoic acid. In some embodiments, about one to ten (e.g., one, two, three, four, five, six, seven, eight, nine, or ten) hydrophilic amino acids may be added to the N-terminus, C-terminus, an internal position, or any combination thereof, of the peptide component of the nerve targeting peptide conjugate to increase solubility. Hydrophilic amino acids include D, E, H, K, N, Q, R, S, T, and G. In some embodiments, the peptide comprises a K, KK, KKK, KKKK (SEQ ID NO:573), G, GG, GGG, or GGGG (SEQ ID NO:574) at the N-terminus or C-terminus.


V. Pharmaceutical Compositions

Disclosed herein, in certain embodiments, are pharmaceutical compositions comprising a nerve targeting peptide conjugate disclosed herein. Pharmaceutical compositions herein are formulated using one or more physiologically acceptable carriers including excipients and auxiliaries which facilitate processing of the active agents into preparations which are used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. A summary of pharmaceutical compositions is found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N. Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins, 1999).


In some embodiments of pharmaceutical compositions comprising the nerve targeting peptide conjugates described herein, the peptide comprises or consists or the amino acid sequence set forth in any one of SEQ ID NOS: 1-557. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS: 1-30. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:31-36. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:37-40. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:41-240. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:241-349. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:350-449. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:450-557.


In embodiments of pharmaceutical compositions comprising the nerve targeting peptide conjugates described herein, the peptide comprises or consists of an amino acid sequence set forth in any one of Formulas (I)-(XLIV) or Subformulas (Ia)-(XLIVa).


In certain embodiments, a pharmaceutical composition disclosed herein further comprises a pharmaceutically acceptable diluent(s), excipient(s), or carrier(s). In some embodiments, the pharmaceutical compositions include other medicinal or pharmaceutical agents, carriers, adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure, and/or buffers. In addition, the pharmaceutical compositions also contain other therapeutically valuable substances.


In certain embodiments, nerve targeting peptide conjugates disclosed herein are delivered to a subject via a drug delivery vehicle or carrier. In some embodiments, a delivery vehicle is made from natural or synthetic materials or both. In some embodiments, a delivery vehicle is a nanoparticle, microparticle, polymeric micelle, nanocapsule, dendrimer, large PEG, nanogel, liposome, fullerene, nanostructured lipid carrier, nanoshell, quantum dot, protein-based nanocarriers (e.g., albumin, elastin, gliadin, legumin, zein, soy protein, milk protein, whey based nanocarriers), organic nanocarrier (e.g., gelatin, dextran, guar gum, chitosan, collagen), polysaccharide based carrier (e.g., dextran, chitosan, pectin), lipid emulsion, or a combination thereof.


In certain embodiments, a pharmaceutical composition disclosed herein is administered to a subject by any suitable administration route, including but not limited to, topical, oral, intrarectal, intravaginal, intranasal, inhalation, parenteral (intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, intravascular, intrathecal, intravitreal, infusion) administration. In certain embodiments, a pharmaceutical composition disclosed herein is administered to a subject is administered locally or systemically. In embodiments, a pharmaceutical composition disclosed herein is administered intravenously.


Formulations suitable for intramuscular, subcutaneous, or intravenous injection include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or vehicles including water, ethanol, polyols (propylene glycol, polyethylene-glycol, glycerol, cremophor and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation. Bioavailability enhancers may include penetration or permeation enhancers. See, e.g., Muheem et al., 2016, Saudi Pharm J. 24, 413-428; Brayden et al 2020, Adv Drug Deliv Rev. 157, 2-36; Ibrahim et al 2020, J Pham. Sci. 28, 403-416. In some formulations, proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Formulations suitable for subcutaneous injection also contain optional additives such as preserving, wetting, emulsifying, and dispensing agents, as are known in the art. In embodiments, formulations suitable for intravenous injection may be prepared in aqueous solutions, such as saline buffers and other physiologically compatible buffers known in the art.


Parenteral injections optionally involve bolus injection or continuous infusion. Formulations for injection are optionally presented in unit dosage form, e.g., in ampoules or in multi dose containers, with an added preservative. In some embodiments, the pharmaceutical composition described herein are in a form suitable for parenteral injection as sterile suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulating agents such as suspending, stabilizing and/or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of an active agent in water soluble form. Additionally, suspensions are optionally prepared as appropriate oily injection suspensions.


In embodiments, a pharmaceutical composition herein is administered orally. Dosage forms suitable for oral administration can be solid or liquid and may include for example, a pill, capsule, troche, tablet, caplet, gel caplet (gel cap), syrup, an aqueous suspension or solution, a chewable form, a swallowable form, a dissolvable form, an effervescent, a granulated form, and an oral liquid solution. In a specific embodiment, the dosage form is a solid dosage form, and more specifically, comprises a tablet or capsule, or is orally administered by a vehicle or carrier as disclosed herein.


In some embodiments, the pharmaceutical composition described herein is in unit dosage forms suitable for single administration of precise dosages. In unit dosage form, the formulation is divided into unit doses containing appropriate quantities of an active agent disclosed herein. In some embodiments, the unit dosage is in the form of a package containing discrete quantities of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. In some embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multiple-dose reclosable containers are used, in which case it is typical to include a preservative in the composition. By way of example only, formulations for parenteral injection are presented in unit dosage form, which include, but are not limited to ampoules, or in multi dose containers, with an added preservative.


In some embodiments, the nerve targeting peptide conjugate is administered via systemic intravenous injection into human patients.


VI. Methods of Use

In another aspect, the present disclosure provides methods of delivering a cargo molecule to a neuron or nerve, comprising contacting the neuron or nerve with the nerve targeting peptide conjugates or pharmaceutical compositions described herein. Such methods may be used to label or identify a neuron or nerve, delivery a drug to a neuron or nerve, or delivery a photosensitizing agent to a neuron or nerve.


In some embodiments, the contacting occurs in vivo. In some embodiments, the contacting occurs in vitro.


In some embodiments of the methods described herein, the nerve targeting peptide conjugates are administered to a subject, as described herein, including humans and mammals (e.g., mice, rats, pigs, cats, dogs, and horses). In some embodiments, subjects are mammals. In some embodiments, subjects are primates. In some embodiments, subjects are humans. In some embodiments, human subjects are pediatric subjects (age 21 years and younger), adult subjects (age 22 years to 65 years), or geriatric subjects (age 65 years and above).


In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS: 1-30. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:31-36. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:37-40. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:41-240. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:241-349. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:350-449. In some embodiments, the peptide comprises or consists of the amino acid sequence set forth in any one of SEQ ID NOS:450-557.


In some embodiments, a peptide comprises or consists of an amino acid sequence set forth in any one of Formulas (I)-(XLIV), including Subformulas (Ia)-(XLIVa).


In some embodiments, the present disclosure provides methods of labeling or identifying a neuron or nerve (or component of either) by contacting a neuron or nerve with nerve targeting peptide conjugate described herein.


In some embodiments, the cargo molecule of the nerve targeting peptide conjugate comprises a fluorescent moiety described herein. In some embodiments, the fluorescent moiety comprises a fluorescent protein, a fluorescent peptide, a fluorophore, or any combination thereof.


In some embodiments, a nerve targeting peptide conjugate is administered in combination (simultaneously, concurrently, or serially) with a fluorescent moiety (e.g., fluorescent moiety that is not conjugated to the nerve target peptide conjugate, or “free” fluorescent moiety). In some embodiments, the fluorescent moiety is a fluorescein, e.g., carboxyfluorescein.


In some embodiments of the methods of labeling or identifying a neuron or nerve described herein, the neuron or nerve is labeled for identification during surgery on the subject. In some embodiments, the surgery is related to a traumatic injury in the subject. In some embodiments, the surgery is related to an infection in the subject. In some embodiments, the surgery is plastic surgery e.g., cosmetic or reconstructive surgery, in the subject.


In some embodiments, the surgery is head, neck, skull base, spinal, prostate, heart, kidney, hand, arm, foot, leg, lower abdominal, or gynecological surgery.


In some embodiments, the surgery is head and neck surgery.


In some embodiments, the surgery is gastrointestinal tract (GI) surgery or genitourinary tract (GU) surgery.


In some embodiments the surgery is cancer surgery. In some embodiments the cancer is selected from the group consisting of prostate cancer, liver cancer (HCC), colorectal cancer, ovarian cancer, endometrial cancer, breast cancer, pancreatic cancer, stomach cancer, cervical cancer, head and neck cancer, thyroid cancer, testis cancer, urothelial cancer, lung cancer, melanoma, testicular germ cell tumors, mesothelioma, and esophageal cancer. In some embodiments, the cancer is prostate cancer.


In some embodiments, the method comprises administering a nerve targeting peptide conjugate disclosed herein to a subject that will undergo surgery. In some embodiments, the method comprises administering a nerve targeting peptide conjugate disclosed herein to a subject that is currently undergoing surgery. In some embodiments, a nerve targeting peptide conjugate disclosed herein is administered to a patient systemically, for example, by intravenous injection. In some embodiments, a nerve targeting peptide conjugate disclosed herein is administered to a patient locally.


Disclosed herein, in certain embodiments, are methods of targeted drug delivery. In some embodiments, a nerve targeting peptide conjugate disclosed herein delivers a drug to a specific target. In some embodiments, a nerve targeting peptide conjugate disclosed herein delivers a drug to a neuron or nerve.


In some embodiments, the drug is an agent that reduces pain (either the perception of pain or activity of a painful stimulant). In some embodiments, the drug is an anesthetic. In some embodiments, the drug is benzocaine; carticaine; cinchocaine; cyclomethycaine; lidocaine; prilocaine; propxycaine; proparacaine; tetracaine; tocainide; and trimecaine; or a combination thereof.


In some embodiments, the drug is an agent that modulates death (e.g., via apoptosis or necrosis) of a neuron or nerve. In some embodiments, the drug is a cytotoxic agent. In some embodiments, the drug is methotrexate (RHEUMATREX®, Amethopterin); cyclophosphamide (CYTOXAN®); thalidomide (THALIDOMID®); paclitaxel; pemetrexed; pentostatin; pipobroman; pixantrone; plicamycin; procarbazine; proteasome inhibitors (e.g.; bortezomib); raltitrexed; rebeccamycin; rubitecan; SN-38; salinosporamide A; satraplatin; streptozotocin; swainsonine; tariquidar; taxane; tegafur-uracil; temozolomide; testolactone; tbioTEPA; tioguanine; topotecan; trabectedin; tretinoin; triplatin tetranitrate; tris(2-chloroethyl)amine; troxacitabine; uracil mustard; valrubicin; vinblastine; vincristine; vinorelbine; vorinostat; zosuquidar; or a combination thereof. In some embodiments, the drug is a pro-apoptotic agent. In some embodiments, the drug is an anti-apoptotic agent. In some embodiments, the drug is selected from minocycline; SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl) 1H-imidazole); PD 169316 (4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole); SB 202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole); RWJ 67657 (4-[4-(4-fluorophenyl)-1-(3-phenylpropyl)-5-(4-pyridinyl)-1H-imidazol-2-yl]-3-butyn-1-ol); SB 220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole); D-JNKI-1 ((D)-hJIP 175_i 57-DPrO-DPrO-(D)-HIV-TAT57-48); AM-111 (Auris); SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one); JNK Inhibitor I ((L)-HIV-T AT48-57-PP-JBD20); JNK Inhibitor III ((L)-HIV-TAT47-57-gaba-c-Jun833-57); AS601245 (1,3-benzothiazol-2-yl (2-[[2-(3-pyridinyl)ethyl]amino]-4 pyrimidinyl) acetonitrile); JNK Inhibitor VI (H2N-RPKRPTTLNLF-NH2) (SEQ ID NO:564); JNK Inhibitor VIII (N-(4-Amino-5-cyano-6-ethoxypyridin-2-yl)-2-(2,5-dimethoxyphenyl)acetamide); JNK Inhibitor IX (N-(3-Cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl)-1-naphthamide); dicumarol (3,3′-Methylenebis(4-hydroxycoumarin)); SC-236 (4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzene-sulfonamide); CEP-1347 (Cephalon); CEP-11004 (Cephalon); an artificial protein comprising at least a portion of a Bcl-2 polypeptide; a recombinant FNK; V5 (also known as Bax inhibitor peptide V5); Bax channel blocker ((+)-1-(3,6-Dibromocarbazol-9-yl)-3-piperazin-1-yl-propan-2-ol); Bax inhibiting peptide P5 (also known as Bax inhibitor peptide P5); Kp7-6; FAIM(S) (Fas apoptosis inhibitory molecule-short); FAIM(L) (Fas apoptosis inhibitory molecule-long); Fas: Fc; FAP-1; NOK2; F2051; F1 926; F2928; ZB4; Fas M3 mAb; EGF; 740 Y-P; SC 3036 (KKHTDDGYMPMSPGVA) (SEQ ID NO:565); PI 3-kinase Activator (Santa Cruz Biotechnology, Inc.); Pam3Cys ((S)-(2,3-bis(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-Cys-(S)-Ser(S)-Lys4-OH, trihydrochloride); Actl (NF-kB activator 1); an anti-DcB antibody; Acetyl-11-keto-b-Boswellic Acid; Andrographolide; Caffeic Acid Phenethyl Ester (CAPE); Gliotoxin; Isohelenin; NEMO-Binding Domain Binding Peptide (DRQIKIWFQNRRMKWKKTALDWSWLQTE) (SEQ ID NO:566); NF-kB Activation Inhibitor (6-Amino-4-(4-phenoxyphenylethylamino)quinazoline); NF-kB Activation Inhibitor II (4-Methyl-Nl-(3-phenylpropyl)benzene-1,2-diamine); NF-kB Activation Inhibitor III (3-Chloro-4-nitro-N-(5-nitro-2-thiazolyl)-benzamide); NF-kB Activation hihibitor IV ((E)-2-Fluoro-4′-methoxystilbene); NF-kB Activation Inhibitor V (5-Hydroxy-(2,6-diisopropylphenyl)-1H-isoindole-1,3-dione); NF-kB SN50 (AA VALLP A VLLALLAP VQRKRQKLMP) (SEQ ID NO:567); Oridonin; Parthenolide; PPM-18 (2-Benzoylamino-1,4-naphthoquinone); Rol06-9920; Sulfasalazine; TIRAP Inhibitor Peptide (RQIKIWFNRRMKWKKLQLRD AAPGGAIVS) (SEQ ID NO:568); Withaferin A; Wogonin; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile); BAY 11-7085 ((E)3-[(4-t-Butylphenyl)sulfonyl]-2-propenenitrile); (E)-Capsaicin; Aurothiomalate (ATM or AuTM); Evodiamine; Hypoestoxide; IKK Inhibitor III (BMS-345541); IKK Inhibitor VII; IKK Inhibitor X; IKK Inhibitor II; IKK-2 Inhibitor IV; IKK-2 Inhibitor V; IKK-2 Inhibitor VI; IKK-2 Inhibitor (SC-514); IkB Kinase Inhibitor Peptide; IKK-3 Inhibitor LX; ARRY-797 (Array BioPharma); SB-220025 (5-(2-Amino-4-pyrimidinyl)-4-(4-fluorophenyl)-1-(4-piperidinlyl)imidazole); SB-239063 (trans-4-[4-(4-Fluorophenyl)-5-(2-methoxy-4-pyrimidinyl)-1H-imidazol-1-yl]cyclohexanol); SB-202190 (4-(4-Fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl) 1H-imidazole); JX-401 (-[2-Methoxy-4-(methylthio)benzoyl]-4-(phenylmethyl)piperidine); PD-169316 (4-(4-Fluorophenyl)-2-(4-nitrophenyl)-5-(4-pyridyl)-1H-imidazole); SKF-86002 (6-(4-Fluorophenyl)-2,3-dihydro-5-(4-pyridinyl)imidazo[2,1-b]thiazole dihydrochloride); SB-200646 (N-(1-Methyl-1H-indol-5-yl)-N′-3-pyridinylurea); CMPD-I (2′-Fluoro-N-(4-hydroxyphenyl)-[1,1′-biphenyl]-4-butanamide); EO-1428 ((2-Methylphenyl)-[4-[(2-amino-4-bromophenyl)amino]-2-chlorophenyl]methanone); SB-253080 (4-[5-(4-Fluorophenyl)-2-[4-(methylsulfonyl)phenyl]-1H-imidazol-4-yl]pyridine); SD-169 (1H-Indole-5-carboxamide); SB-203580 (4-(4-Fluorophenyl)-2-(4-methylsulfmyl phenyl)-5-(4-pyridyl) 1H-imidazole); TZP-101 (Tranzyme Pharma); TZP-102 (Tranzyme Pharma); GHRP-6 (growth hormone-releasing peptide-6); GHRP-2 (growth hormone-releasing peptide-2); EX-1314 (Elixir Pharmaceuticals); MK-677 (Merck); L-692,429 (Butanamide, 3-amino-3-methyl-N-(2,3,4,5-tetrahydro-2-oxo-1-((2′-(1H-tetrazol-5-yl)(1,1′-biphenyl)-4-yl)methyl)-1H-1-benzazepin-3-yl)-, (R)-); EP1572 (Aib-DTrp-DgTo-CHO); diltiazem; metabolites of diltiazem; BRE (Brain and Reproductive organ-Expressed protein); verapamil; nimodipine; diltiazem; omega-conotoxin; GVIA; amlodipine; felodipine; lacidipine; mibefradil; NPPB (5-Nitro-2-(3-phenylpropylamino)benzoic Acid); flunarizine; erythropoietin; piperine; hemin; brazilin; z-V AD-FMK (Benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone); z-LEHD-FMK (benzyloxycarbonyl-Leu-Glu(OMe)-His-Asp(OMe)-fluoromethylketone) (SEQ ID NO:569); B-D-FMK (boc-aspartyl(Ome)-fluoromethylketone); Ac-LEHD-CHO (N-acetyl-Leu-Glu-His-Asp-CHO) (SEQ ID NO:569); Ac-IETD-CHO (N-acetyl-Ile-Glu-Thr-Asp-CHO) (SEQ ID NO:570); z-IETD-FMK (benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone) (SEQ ID NO:570); FAM-LEHD-FMK (benzyloxycarbonyl Leu-Glu-His-Asp-fluoromethyl ketone) (SEQ ID NO:569); FAM-LETD-FMK (benzyloxycarbonyl Leu-Glu-Thr-Asp-fluoromethyl ketone) (SEQ ID NO:571); Q-VD-OPH (Quinoline-Val-ASp-CH2-O-Ph); XIAP; cIAP-1; cIAP-2; ML-IAP; ILP-2; NAIP; Survivin; Brace; IAPL-3; fortilin; leupeptine; PD-150606 (3-(4-Iodophenyl)-2-mercapto-(Z)-2-propenoic acid); MDL-28170 (Z-Val-Phe-CHO); calpeptin; acetyl-calpastatin; MG 132 (N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(1S)-1-formyl-3-methylbutyl]-L-leucinamide); MYODUR; BN 82270 (Ipsen); BN 2204 (Ipsen); AHLi-11 (Quark Pharmaceuticals), an mdm2 protein, pifithrin-α (1-(4-Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl)ethanone); trans-stilbene; cis-stilbene; resveratrol; piceatannol; rhapontin; deoxyrhapontin; butein; chalcon; isoliquirtigen; butein; 4,2′,4′-trihydroxychalcone; 3,4,2′,4′,6′-pentahydroxychalcone; flavone; morin; fisetin; luteolin; quercetin; kaempferol; apigenin; gossypetin; myricetin; 6-hydroxyapigenin; 5-hydroxyflavone; 5,7,3′,4′,5′-pentahydroxyflavone; 3,7,3′,4′,5′-pentahydroxyflavone; 3,6,3′,4′-tetrahydroxyflavone; 7,3′,4′,5′-tetrahydroxyflavone; 3,6,2′,4′-tetrahydroxyflavone; 7,4′-dihydroxyflavone; 7,8,3′,4′-tetrahydroxyflavone; 3,6,2′,3′-tetrahydroxyflavone; 4′-hydroxyflavone; 5-hydroxyflavone; 5,4′-dihydroxyflavone; 5,7-dihydroxyflavone; daidzein; genistein; naringenin; flavanone; 3,5,7,3′,4′-pentahydroxyflavanone; pelargonidin chloride; cyanidin chloride; delphinidin chloride; (−)-epicatechin (Hydroxy Sites: 3,5,7,3′,4{circumflex over ( )}; (−)-catechin (Hydroxy Sites: 3,5,7,3′,4); (−)-gallocatechin (Hydroxy Sites: 3,5,7,3′,4′,5) (+)-catechin (Hydroxy Sites: 3,5,7,3′,4{circumflex over ( )}; (+)-epicatechin (Hydroxy Sites: 3,5,7,3′,40; Hinokitiol (b-Thujaplicin; 2-hydroxy-4-isopropyl-2,4,6-cycloheptatrien-1-one); L-(+)-Ergothioneine ((S)-a-Carboxy-2,3-dihydro-N,N,N-trimethyl-2-thioxo-1H-iniidazole4-ethanaminium inner salt); Caffeic Acid Phenyl Ester; MCI-186 (3-Methyl-1-phenyl-2-pyrazolin-5-one); HBED (N,N′-Di-(2-hydroxybenzyl)ethylenediamine-N,N′-diacetic acid*H2O); Ambroxol (trans-4-(2-Amino-3,5-dibromobenzylamino)cyclohexane-HCl; and U-83836E ((−)-2-((4-(2,6-di-1-Pyrrolidinyl-4-pyrimidinyl)-1-piperzainyl)methyl)-3,4-dihydro-2,5,7,8-tetramethyl-2H-1-benzopyran-6-ol>>2HCl); β-1-5-methyl-nicotinamide-2′-deoxyribose; /3-D-1′-5-methyl-nicotinamide-2′-deoxyribofuranoside; /3-1′-4,5-dimethyl-nicotinamide-2′-de-oxyribose; /3-D-1′-4,5-dimethyl-nicotmamide-2′-deoxyribofuranoside; 1-Naphthyl PPI (1-(1,1-Dimethylethyl)-3-(1-naphthalenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); Lavendustin A (5-[[(2,5-Dihydroxyphenyl)methyl][(2-hydroxyphenyl)methy 1]amino]-2-hydroxybenzoic acid); MNS (3,4-Methylenedioxy-b-nitrostyrene); PPI (1-(1,1-Dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); PP2 (3-(4-chlorophenyl) 1-(1,1-dimethylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); KX1-004 (Kinex); KX1-005 (Kinex); KX1-136 (Kinex); KX1-174 (Kinex); KX1-141 (Kinex); KX2-328 (Kinex); KX1-306 (Kinex); KX1-329 (Kinex); KX2-391 (Kinex); KX2-377 (Kinex); ZD4190 (Astra Zeneca; N-(4-bromo-2-fluorophenyl)-6-methoxy-7-(2-(1H-1,2,3-triazol-1-yl)ethoxy)quinazolin-4-amine); AP22408 (Ariad Pharmaceuticals); AP23236 (Ariad Pharmaceuticals); AP23451 (Ariad Pharmaceuticals); AP23464 (Ariad Pharmaceuticals); AZD0530 (Astra Zeneca); AZM475271 (M475271; Astra Zeneca); Dasatinib (N-(2-chloro-6-methylphneyl)-2-(6-(4-(2-hydroxyethyl)-piperazin-1-yl)-2-methylpyrimidin-4-ylamino) thiazole-5-carboxamide); GN963 (trans-4-(6,7-dimethoxyquinoxalin-2ylamino)cyclohexanol sulfate); Bosutinib (4-((2,4-dichloro-5-methoxyphenyl)amino)-6-methoxy-7-(3-(4-methyl-1-piperazinyl)propoxy)-3-quinolinecarbonitrile); or combinations thereof.


In some embodiments, the drug is an agent that reduces undesired neuron or nerve impulses. In some embodiments, the drug reduces one or more symptoms of dyskinesia or synkinesia. In some embodiments, the drug is carbamazepine, oxcarbazepine, phenytein, valproic acid, sodium valproate, cinnarizine, flunarizine, or nimodipine, or combinations thereof.


In some embodiments, the drug is an agent that promotes regeneration of neuron or nerve tissue. In some embodiments, the drug is a growth factor. In some embodiments, the drug is selected from brain-derived neurotrophic factor (BDNF); ciliary neurotrophic factor (CNTF); glial cell-line derived neurotrophic factor (GDNF); neurotrophin-3; neurotrophin-4; fibroblast growth factor (FGF) receptor; insulin-like growth factor (IGF); or a combination thereof.


Disclosed herein, in certain embodiments, are methods of delivering a photosensitizing agent to a human neuron or nerve comprising: contacting the human neuron or nerve with a nerve targeting peptide conjugate comprising (a) a nerve targeting peptide comprising the amino acid sequence of any one of SEQ ID NOS: 1-557, and (b) a photosensitizing agent. In some embodiments, the method further comprises exposing the human neuron or nerve with a light source that activates the photosensitizing agent, wherein the activated photosensitizing agent induces ablation or killing of the human neuron or nerve. Upon exposure to a specific wavelength of light, a photosensitizing agent reacts with molecular oxygen to produce singlet oxygen, which is cytotoxic. In certain embodiments, a photosensitizing agent is a porphyrin, chlorin, or dye. Examples of photosensitizing agents include porphyrin, protoporfin IX, purlytin, verteporfin, HPPH, temoporfin, methylene blue, photofrin, protofrin, hematoporphyrin, Talaporfin, benzopophyrin derivative monoacid, 5-aminileuvolinic acid, Lutetium texaphyrin, metallophthalocyanine, metallo-naphthocyaninesulfobenzo-porphyrazines, metallo-naphthalocyanines, zinc tetrasulfophthalocyanine, bacteriochlorins, metallochlorins, chlorine derivative, Tetra(m-hydroxyphenyl)chlorin (mTHPC), pheophorbide, dibromofluorescein (DBF), IR700DX, naphthalocyanine, and porphyrin derivatives.


Human neuron or nerve targeting molecules comprising a photosensitizing agent as disclosed herein can be used in methods of localized nerve killing in a subject. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating chronic pain (e.g., back, neck, or joint pain) in subject. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating prostate cancer in a subject. Autonomic innervation may contribute to prostate cancer growth and metastasis by light induced ablation of local autonomic nerves. Thus, local autonomic nerves may be a viable target for prostate cancer therapy. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating renovascular hypertension in a subject by light induced ablation of sympathetic nerves in the renal vessels. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating excessive sweating. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating cardiac arrhythmias. In some embodiments, human neuron or nerve targeting molecules comprising a photosensitizing agent are used for treating pathological muscle spasms (e.g., Meige syndrome, hemifacial spasm, torticollis).


In some embodiments, a first nerve targeting peptide conjugate is administered in combination (simultaneously, concurrently, or serially) with a second nerve targeting peptide conjugate. In yet further embodiments, the cargo molecule of the first nerve targeting peptide conjugate and the cargo molecule of the second nerve targeting peptide conjugate are each independently selected from a fluorescently moiety, drug, photosensitizing agent, neurotrophic factor, and radiolabel. In some embodiments, the cargo molecule of the first nerve targeting peptide conjugate and the cargo molecule of the second nerve targeting peptide conjugate are the same or different.


EXAMPLES
Example 1: Phage Display Library Construction, Screening, and Identification of Nerve-Binding Peptides

Screens of phage display libraries were used to identify peptides with selective binding for nerves versus muscle and therefore may be useful for systemic in-vivo labeling of nerves during fluorescence assisted surgery, as well as other nerve targeting applications.


Construction of Library

Several m13 phage libraries were constructed expressing randomized variants of a parent peptide (QVPWEEPYYVVKKS-SGGL; SEQ ID NO: 576). In particular, the first 14 amino acids of the parent peptide were randomized using degenerate oligonucleotide synthesis to obtain a diverse set of peptides with a mutation rate of about 50% per residue.


To accomplish this, synthesis was carried out by the XXK codon design approach, where X is any base and K is G or T. To bias synthesis towards the parent sequence, synthesis was carried out at each X position with a mixture of about 70%-85% of the parent nucleotide and 15-30% of the remaining 3 nucleotides. The resulting molecules were inserted into fADL-1e (Antibody Design Laboratories), which is a type 3 phage display vector having a cloning site on the N-terminal side of the full-length gene III protein. The fusion proteins are directed to the periplasm by the PelB leader peptide, which is inserted in the vector in place of the wild-type gene III protein. During the phage assembly process, the peptides are displayed on the surface of the nascent phage particles.


Collectively, the phage display libraries used in the screens comprised about 600 million peptides, with sequencing analysis confirming a mutation rate of about 50% per residue.


Screening and Sequencing

Phage libraries were independently processed, as follows, with five biological samples: two independent samples of excised human nerve tissue, one sample of a human nerve extract, and two independent samples of human muscle tissue, as follows:


Excised Human Nerve Tissue: For each of the two independent samples, phage were processed through multiple cycles of selection, with representative phage being isolated and sequenced after each selection cycle. Specifically, for selection using human nerve tissue, a sample of nerve muscle tissue (2 mm by 2 mm) was washed prior to mixing and incubation with a phage library. After incubation, the mixture (containing mostly intact nerves with phage particles that had variable affinity for nerves) was centrifuged and the nerve pellet washed multiple times with PBS to remove phage with little or no affinity for nerve tissue. The pellet was homogenized and plated for titering and reamplification. Phage that were bound at each round were sequenced. NextGen sequencing of pools after 2, 3 and 4 rounds of selection were used to isolate candidate sequences that were further analyzed using bioinformatics.


Human Nerve Extracts: Phage libraries were incubated for up to 24 hours at 4° C. in 96 well plates that had been coated with a human nerve extract. Plates were then washed with PBS and phage were eluted using 0.2M glycine pH 2.0. Phage supernatants where then neutralized with Tris pH 9.0 prior to plating on TG1 bacteria and LB agar plates. Colonies were counted to determine titer followed by DNA preparation of pools of phage and NextGen sequencing. After rounds of selection, phage were pooled and amplified for iterative selection. Phage that eluted by 0.2M glycine pH 2.0 (the nerve binders) at each round were sequenced. NextGen sequencing of pools after 2, 3 and 4 rounds of selection were used to isolate candidate sequences that were further analyzed using bioinformatics.


Human Muscle Tissue: For each sample, phage were processed through multiple cycles of selection, with representative phage being isolated and sequenced after each selection cycle. Specifically, for selection using human skeletal muscle tissue, a sample of skeletal muscle tissue (2 mm by 2 mm) was washed prior to mixing and incubation with a phage library. After incubation, the mixture (containing mostly intact muscle with phage particles that had variable affinity for muscle) was centrifuged and the muscle pellet washed multiple times with PBS to remove phage with little or no affinity for muscle tissue. The pellet was then homogenized and plated for titering and reamplification. Phage that were bound at each round were sequenced.


NextGen sequencing of pools after 2, 3 and 4 rounds of selection were used to isolate candidate sequences from each of the five samples. These five data sets were then analyzed as described in the next section.


Processing of Peptide Sequences

A python script was used to translate, trim, and filter the peptide sequences:


Peptide and vector sequences were translated starting at the first start codon in the nucleotide sequence. The leader sequence, comprising the first 22 amino acids, was trimmed from the translated protein, the following 14 amino acids were saved as the displayed binding peptide, and the remaining amino acids after the peptide (derived from the vector) were also trimmed. Trimmed peptides with identical sequences were combined and quantified according to their frequency (count).


The script then removed peptide sequences that were likely artifacts (or otherwise not useful) by filtering out: 1) sequences without a start codon, 2) sequences with more than two changes in the leader sequence, 3) C-terminal sequences (after the peptide) that did not start with amino acids “SGGGG” (SEQ ID NO:575) or were of incorrect length except if they had an extra or missing “SGGGG” (SEQ ID NO:575), 4) peptides that had a single or an odd number of Cysteines, 5) peptide sequences that were truncated by stop codons, and 6) peptide sequences that occurred less than six times in a dataset.


Table 1 summarizes the number of sequences corresponding to these processing steps for each of the 5 data sets:















TABLE 1







ABMR2
ABMCR3
ABNCR2
ABNCR3
ABNPR2



(Muscle2)
(Muscle3)
(Nerve2)
(Nerve3)
(Plated Nerve2)





















seqs
52215
25520
52632
26498
34022


no_start
9
3
21
5
10


incorrect_leader
690
533
592
525
602


length_error
1887
1184
1840
1129
1464


C-terminal error
2715
2074
2647
1895
2259


odd cysteines
2773
395
2902
1134
1441


has stop codon
318
263
268
218
241


combined peptide
24106
4215
33093
6629
10484


total


unique peptides
4307
258
2064
366
2809


(count of at least 6)









Assessing Nerve Versus Muscle Binding

Following processing, an R script was written to merge the unique peptide sequences (with counts 6 or greater) from the 5 different data sets to analyze the overlap between nerve and muscle binding activity.



FIG. 1A shows a Venn diagram, showing a high resolution overlap between the five data sets. Collectively, the analysis revealed 2679 unique peptides binding to nerve versus muscle, 3769 unique peptides binding to muscle versus nerve, and 681 unique peptides binding to both muscle and nerve. Peptide sequences within each group were ranked by unique sequence count, that is, their relative enrichment in each set.


Obtaining a Representative Set of Nerve-Selective Peptides

A representative group of 557 nerve-specific peptides was obtained from all 7 subsets of binding data, as shown in FIG. 1B. For the single Nerve2, Nerve3, and Plated Nerve2 (pn2) subsets, the number of counts for each group was set at a threshold giving rise to at least 100 peptides. As highlighted in gray in FIG. 1B, this resulted in 109 unique nerve 2 peptides (having at least 10 counts); 100 unique nerve 3 peptides (having at least 77 counts); and 108 plated nerve 2 peptides (having at least 36 counts). As highlighted in yellow in FIG. 1B, the representative group also included 6 unique peptides overlapping between nerve2 and nerve3, 200 unique peptides overlapping between nerve2 and plated nerve2 (at least 60 counts), 4 unique peptides overlapping between nerve3 and plated nerve2. Finally, as highlighted in green in FIG. 1B, the representative group also included 30 unique peptides common to the nerve2, nerve3, and plated nerve2 sets.


Tables 2A-2G show the peptide sequences and corresponding counts of the 7 groups making up the representative sample of 557 peptides.











TABLE 2A





SEQ ID
Nerve2_Nerve3_



NO: #
Plated Nerve 2
Count







 1
TIPYAEPYYVVRKY
711





 2
DFTMQEPYYAEKKT
593





 3
HVTWQEQYYVLKTP
371





 4
HVPGVDPYSVVKKF
368





 5
HVAFEQPDNVLVLY
357





 6
QVPTEVQFYLVKEP
337





 7
QLPSEQPCCVVKKY
329





 8
QDPWVTRYYIVKKS
304





 9
QVPAKEPYNFMKKS
299





10
YLTWEEPEHVNKQS
272





11
QVPRQELYYVSHQF
245





12
QHPFEQSFYVVNKF
227





13
DILLDELRFAEKHG
203





14
YNLWDAPSYLVKNS
201





15
HVAGGERYFSVKES
185





16
HLPSDQPLSVTKIS
173





17
GFLGEEPYHFLAKR
167





18
LRSTADAHYTLKKS
152





19
ATPCSQPCAVVPKF
142





20
QYAWEEPYYLEKKL
140





21
HLPVEYPFYVFKNS
131





22
RGSLEGPFKLGKTH
130





23
QVSSQEPNFIVKNN
116





24
HFSAYEQYYVLRNS
111





25
YVIKYAPYYAVTRS
 93





26
HLPEEELYTIVRKT
 85





27
QDPCVWPYNGVKKC
 79





28
HVHWDATYYAVTNS
 77





29
QFPYDHKNYVVKYS
 69





30
RFHMVGPYNVATTS
 68


















TABLE 2B





SEQ ID




NO: #
Nerve2_Nerve3
Count







31
HLPWEGQLNIGNKS
357





32
DDLWQDLYGVVKKS
106





33
NLAWAESKFLLEKI
 71





34
QLRYQEHYYLGYNS
 56





35
YVPWQEPYYVQKNL
 49





36
LVPWDETHKVVQRH
 14


















TABLE 2C





SEQ ID
Nerve3_Plated



NO: #
Nerve 2
Count







37
HVALEQPYYAVKES
342





38
QVQCKEQYYGVTNC
195





39
QALYAEPAYVSKKF
 19





40
HVSMDERYYVVKKT
 17


















TABLE 2D





SEQ ID
Nerve2_Plated



NO: #
Nerve2
Count







 41
HSYREEPYYFGKRS
632





 42
QHTWKEPYNGQKRT
577





 43
QLPMGGPYERVNNT
380





 44
SVPKVHPYYVVNNS
333





 45
QVPTGEPLYPVQKT
323





 46
QLPSYQAHDVMKVH
308





 47
QIPSQRPSSVGAIA
292





 48
QFPGDERYYLATST
283





 49
HVSSQEPYYVLRNS
252





 50
RLPSDDHNTFGTRS
249





 51
QLHGDELLSVVKNP
232





 52
ATEWEESYFHVKKF
223





 53
RVPWEQSSWVVKRS
220





 54
QIANKEPHTYVENS
213





 55
QIRSEVTHSVVRES
213





 56
QHTLNHPYSVVKTS
205





 57
QVPSHELYFRPKES
204





 58
EVPGGVPYYPMKKF
201





 59
HVSGGQWVHGQKNS
200





 60
HVPSKQMNSLAKKS
196





 61
QIHFGEPNYVVKKS
194





 62
QFPWEGSYYTAKQS
193





 63
QIPWGEAYQAEKIT
191





 64
VVPSNAQHLAPNMS
191





 65
HVAEAESYNVVKKT
187





 66
QIPCEGPYSGVQMC
186





 67
HITGGVPYPVVRDS
185





 68
HVLWEDSSYHVTHH
179





 69
HVTYDEQKYFVKKS
179





 70
QVPSVQPYYIGKNT
175





 71
QARFEQPYSGLRDS
167





 72
LIPSAYPHSLWSKS
166





 73
QLRSNEAYHVVKKS
166





 74
QTQMQEPKYGVSHS
163





 75
DVRWDEPYYVDEKI
161





 76
HVPRVEPYNVVTNY
158





 77
QFPGQALSFVHEKS
152





 78
HFPSKALNNTAKNT
151





 79
QFPPEQPYGLANKS
150





 80
HVPWAGRHYVAKHS
149





 81
RVAIDELYLIAHKS
147





 82
QIPEQQPYNVFKKS
142





 83
SIPGEDSLYAVTEH
142





 84
QCPSEQPNYVCKIF
140





 85
HGQTQEPSYVTVYY
139





 86
HVHWQVPSNIVRNS
139





 87
QLPWYEADYRMEYY
139





 88
QLPSQAQNYYVTAS
137





 89
QVTSEEHYFAVKKF
136





 90
TSLSGAPFHNANTF
134





 91
HDASIALYDPVLMS
133





 92
QLPGDVTYNLGHKS
133





 93
QPTWEQPYHVDRNS
131





 94
HIPFPGLYSNLDQS
127





 95
DHPWDVRHYAHKKS
126





 96
DVPWKESFNIVENS
126





 97
QFARTAPYYVQTEY
125





 98
QNPWQKPYTVMKTS
123





 99
HLSFEEPLYGVKKS
122





100
QFPSGHRVYTGKMT
122





101
YNLYEQPYSVLKRS
122





102
HLPSQVTHYVITSF
115





103
QVPSHEPFYLGKKY
114





104
RVTQGSPVYGENNA
114





105
QIPMEETYSLRKMS
113





106
QIPWEAPSYGATIS
113





107
HVGWEQPYYVLRKS
112





108
DFATEERRYAVPKY
111





109
DVPLVKPHYGAVDS
111





110
QFPSEASHYVVRNS
111





111
HSRLHETSYVFNKP
110





112
HVQRQETDYAAKRI
110





113
NVLSEEPSSRDANV
110





114
HLPFEAPYYAVKRS
109





115
QLPGEEPTYVGSNS
109





116
EVPVGDLSYVGKKY
108





117
QFPLVQPNYLVEMP
108





118
HVPGQDSYNVVNKF
107





119
HVTYTEASVYKRTF
107





120
QLEWEEPHYAARKS
107





121
HYQSVKSYSVQKKI
105





122
QLTSAEASYVGKQS
105





123
HRPWEEPYYVAKQS
104





124
LFASENPYNVKSKS
103





125
SVPGEDPSQFGKTP
103





126
HVPSEEPYKLDEQS
102





127
QFRIEDSYDVANQS
102





128
QIYWHESNYLVTKS
102





129
HIPGDETSYVMKKY
100





130
QSLSEEPQYVVTPS
 98





131
HILLEEPSNVLNKR
 96





132
TLPVYVPYYVVNNT
 96





133
HMRSEQPYYGGKVS
 95





134
EVPRTERNFVVKNT
 94





135
HGAWEETHYVVIQS
 94





136
HGRSVESYDLVNKR
 94





137
QVTWGEPQYVVYKS
 94





138
QLPSGEPYSFVKES
 93





139
DVPLEDTYYRVKKY
 91





140
HVPFDKSSHLVKKY
 90





141
QFAWDSPYSRGKKT
 90





142
HVTWQQTKYVTNRS
 89





143
RFPLEEPNYFQTKS
 89





144
HVTWGKTYHLVTDS
 88





145
LVPWEEPGYFONTS
 88





146
QYPREEPRHALKMS
 88





147
NVTFEQSSYVVTSP
 87





148
YVTSESPYYVHGQP
 87





149
HVSGTKPYHLVKKS
 86





150
EDVLHNAYHVETQF
 85





151
QFAWEHRDYVLQRS
 85





152
QVLWEERLYVVKNS
 83





153
RSTWEQSYTIVKES
 83





154
QMASKEPNYPVKNS
 82





155
QNPAEEQYIVWNNS
 82





156
SHPWEKQYDVFNNS
 81





157
DVPWEEPYHVGNWR
 80





158
HVASEQPYYGVNRP
 80





159
HVLGEEPYYFVQHV
 80





160
QEPWDEFSYNVNAL
 80





161
QFRSGEAYVVVQTS
 80





162
QNAFSKPYNGVKPS
 80





163
QIESGEPYHDQKHP
 79





164
QVRWPEQYYVVNMG
 79





165
QYAREEPYYFMSYK
 79





166
QIPCQSDHNVVTNC
 78





167
DRLSDEPFKGMTHS
 77





168
DVRWSDPHDRITKS
 77





169
HLPWEGPEYLGKKS
 77





170
HVPRDEPQSFVHRS
 77





171
QFSSAEAYTVVNES
 77





172
QIPSHSPYYALNSS
 77





173
QQRLSASYQHVIKS
 77





174
HFEGEDLYDRVKSR
 76





175
HVTLEKPYDVATKS
 76





176
QLPGEHPYYPIKKP
 76





177
EALWYKHYYGVHKT
 75





178
HVVRHYPPHFVHKS
 75





179
LIPSEERSYAVINT
 75





180
QHTHKQPFNVVKNY
 75





181
VVPSQGTYHVGMTS
 75





182
AVAWERPYYVVNHS
 74





183
DVLLARTYYFGEFP
 74





184
HFAGQEHLHFVEIS
 74





185
LFAWAEANNVMKNS
 74





186
SITNKQSVYVLKKS
 73





187
HVQSANPYYDVRNS
 72





188
HVSFDEPYYYVKKS
 72





189
QDYWAKPSYIVKHS
 72





190
QISLTEATLVVRKS
 72





191
DVLWEEPRYFVNNS
 71





192
HAHGQETYFLVKQS
 71





193
HFALQDPYQVFKKT
 71





194
QYPAERPTFDVTNA
 71





195
SLPMEERYYAVRTA
 71





196
DVEWEEPYYFLEKS
 70





197
DVTWEEPYYAVKNS
 69





198
EHPRVESVYLFPHT
 69





199
HDSLDLHYYAMHES
 69





200
QCPLEESCFDGQKS
 69





201
QLTWPKQNYTMTQS
 69





202
QLPWGAPYYVVKNS
 68





203
RLPSEEPYTVAKRP
 68





204
DFPWEQRSSTVKQY
 67





205
DHTRVHPYYDFEIP
 67





206
HATLQEPYFVVNHY
 67





207
HDTWKGPYYPANKS
 67





208
QLPSQEPYNVQRKS
 67





209
SVTRYDLSTDVTKS
 67





210
ESQWEQPSDVFKVV
 66





211
KLPLEEPFYIVTPS
 66





212
QIRWEDASYVEKTS
 66





213
QVPWKGQYYVVASS
 66





214
HGPTDGPYYVSKKS
 65





215
HILRQEPYYVLTNY
 65





216
HSHGQELFYVVTNT
 65





217
HVPWGEPEALVKKS
 65





218
QPPREQPYHIMKNS
 65





219
QTSWAGPHDRVTKA
 65





220
QVPCEEPWCLVKVN
 65





221
QVPWQDPVYDVNNA
 65





222
YVKTVNPDSVLKIS
 65





223
HVHFTDPHYVMKKS
 64





224
HVPLQGPYYVKQKP
 64





225
NVEWEIPYYFVKKF
 64





226
QFAWVQPYYFAKKS
 64





227
QHPVKERDEFWKES
 64





228
QIPWGETTAGIKKL
 64





229
EFSREQPNDFLNKS
 63





230
HIPFAEHDGVVPIS
 63





231
HVTSHEARYLAQNS
 63





232
EFTWEGLSEGVNEP
 62





233
HVSSEYPYYVVKHT
 62





234
HVPAEPLVYENQKS
 61





235
LIVSQPSYLAVQYS
 61





236
QLPLAKPNSVVATS
 61





237
RLAWEEPHYVVKKS
 61





238
KFTTQDSYYFGTQT
 60





239
NVTSREPYHLLEKS
 60





240
QLPFIGPYHVAKKS
 60


















TABLE 2E





SEQ ID




NO: #
Nerve2
Count







241
HDASGVSYYDRRES
21





242
HVGLPEPHHVMIHS
20





243
EVPSLQPYHGGKKS
19





244
QIPSQEPYHRFKES
19





245
QIPGEAPYSFGKKY
18





246
QNPWHYPYSLVSTA
17





247
DVPFRLWNFAVVDS
15





248
HFAWVVPNDAVKAS
15





249
HFSWEERYRVVHKS
15





250
RVPSDEPYSFLRQS
15





251
TQRWAEPYHVFHKS
15





252
HVLRHEPYYVVAQA
14





253
LLPSDEAHYIVKHS
14





254
NVLSEQPGYVMEYL
14





255
QLPWTNPFHVVNKY
14





256
QYSLDESPYGAKKS
14





257
YVPGAAPIGIVHKS
14





258
HFAWEEVFHVKKNF
13





259
HSPSKESHYVLENM
13





260
HVPFGGTYSVEGES
13





261
HYRCVGSCYVVAKS
13





262
LFRSQAQNVLVISH
13





263
QFPGSELTVSVIQV
13





264
QFSSQEAHYELNTP
13





265
QSPWDEPYYGVLMA
13





266
QSTPEERYHRVIQA
13





267
QVPLGAPFYAVRKS
13





268
YERWEAPFYDVQKS
13





269
HVAWNEPHHVLLQI
12





270
HVLWEDPYYVVRMP
12





271
HVQLREHYFVVYSS
12





272
HVSSEAPYYVLNKS
12





273
LFPLDETYYVAKVS
12





274
QALLEEKYTALKKS
12





275
QFPWEQRNYDVKKY
12





276
QIPLDARSYLLKRP
12





277
QIQREEPFYVLKLY
12





278
QLPSKASNSVQQTF
12





279
QLPWEKPYYDVEVT
12





280
QNPGKSPYYVVHKS
12





281
QVWGEVPLSGAKRF
12





282
RVPSAAAYYGAKIS
12





283
VFSSDSQYLFVPNF
12





284
DRPVEVPSDFVNNS
11





285
DVLGIERYYAVKDP
11





286
HHPGVGPFHVVKRS
11





287
HLSQYEPYDPVDKS
11





288
HVAWDGAYNLVQTT
11





289
HVPGQDPYDVQATY
11





290
HVPLEGSYLDDINS
11





291
HVTLAEPHLLLGKS
11





292
KFSLVHPEYYGKET
11





293
NLPGVEQYNLPNSS
11





294
NVPWQVPYSVLKMP
11





295
QFAGEVPYYFATDL
11





296
QHPMDPTYHVVIKA
11





297
QILSEEPYYRVDMP
11





298
QIPLAYSNYVVNKY
11





299
QITFEVPCCLVRKG
11





300
QLGLDKLNHLVKKS
11





301
QSSSVEPYYVLQNS
11





302
QVACEQPYYNVNCS
11





303
RLPEAEPYYLMTQP
11





304
RVPGERQYDALNLS
11





305
YVPLQEPYYVVKQS
11





306
ESALKEPHQVLKHS
10





307
HAPADQLYYPVWKP
10





308
HFPWEEPYHVAKRS
10





309
HFQSEEPYHIPHKH
10





310
HHPLQEPYYVVEKS
10





311
HMPGREPYYVVTTS
10





312
HVAWKGPYYVLETH
10





313
HVHGAPSYSVLKHS
10





314
HVLWVEPYYVINKY
10





315
HVPRVDHLYVETKS
10





316
HVPSGERYYVVKTI
10





317
HVPYVGAYNGVKKS
10





318
HVQWEGPYYDVKKS
10





319
HVRGAELSYVLTKS
10





320
HVTWGEPYQVLNKS
10





321
KLLTEAPYMFRTQA
10





322
KLPWQESYDIVRKS
10





323
KLQWQDPDYVVKNS
10





324
KQLGVLPDYVRKNT
10





325
LAPFEERSYALKSP
10





326
LASWDEPSSVLRES
10





327
LLPPDEPYYPMKKS
10





328
NLQWEQPSYVAKDY
10





329
NVPWSEQYNALKTS
10





330
QFASSHTDYVADKS
10





331
QIQWQGSYNDLNKS
10





332
QISSEEPTSLLDRP
10





333
QKPTGASYYGTKPS
10





334
QLAFDGPSPILTTS
10





335
QLHWGQPYSILNKS
10





336
QLPCEVPCYLVQYS
10





337
QLPLQYRYYFVDQS
10





338
QLPWEGKYQLGKKR
10





339
QLRWHEPYHVVNRY
10





340
QSQCAGPGSAYKKC
10





341
QTLAGETNSVVLKF
10





342
QTQCEEPCYVVKFA
10





343
QVPSEEQFYAAVKS
10





344
QVPSENPFRVLKNL
10





345
QVRYVASSDLVRIP
10





346
QVSLQEPGDVLKEF
10





347
QVSREEPSGVVNKP
10





348
RLPSEALYYLVKRS
10





349
YVPVEGIQHVVEKS
10


















TABLE 2F





SEQ ID




NO: #
Nerve3
Count







350
HVRCEEPYCHVRKS
755





351
ELPWQKRKDVMKYP
638





352
QMPWEVPYYVVKRS
548





353
DDIWEVPKYRPKNS
475





354
QIASEERDNIVKKS
462





355
RVPWQEPYYVVLKP
419





356
QLPGYEPYYVGTVS
412





357
RIPSVGPYGVVKTF
391





358
QDVWAEPYYAGKQP
390





359
HVAWQEPRYFVKKS
359





360
YLFAEELAYNLTKS
359





361
QRPWEQAYFVVRST
357





362
FVSREEPYYVVKKS
341





363
DVAREKPYYDMKKS
335





364
QFPSVGSYYVVPQS
331





365
HVVWESSHFVVIDS
329





366
QLRLGDLYNVMTKE
328





367
QFQWQELLNVVKDS
327





368
HVPLQEADYVLKKF
315





369
QFPLYDPYEVMPKS
310





370
HFRWDEPYYIANKA
300





371
RDRHEDSKFVMNHS
290





372
QVPFAHPYYVVNMS
288





373
HVRWGEPSYVLKNF
272





374
QGQTETHSHDAKRY
270





375
QLRFEGPDNPGVQS
269





376
QLPWEVPYSVVRQS
267





377
HVPLPGPYFVLKNY
263





378
HVKRIETYYVFTKS
258





379
RIHLYQPYSILKNY
258





380
QFTSEETHYVVNKS
257





381
QHPWEDPYKIFNKY
240





382
DVPWPNAYNFGIKS
238





383
DVPFEHSYYGEQQY
237





384
EFLGEAPYYVVSRS
236





385
HVSGEEPYTVGNTS
234





386
QFQLDQSYYSFNTL
231





387
QILRVHPDYVVVNP
231





388
HFPWDEAYLVVKNP
230





389
HFSSQESNSGGKKW
230





390
HISGEDPYFILQNS
212





391
TFQYEQPNLVVKNA
207





392
LLAWDEQDYFLRKT
199





393
HINWDKRYAVVITY
197





394
HFPSEYPFDLVSKS
196





395
QFLWQQPDHVRRKS
196





396
QIVLNAPYYVVKTS
189





397
TVPAHNPYYLVKQT
185





398
QDPSVDPYYVDRSS
181





399
QFHCHGACYIVKNS
180





400
RLPQHESYIVGKIS
174





401
RVYWIDPYYIVNES
174





402
QIPLKTPYNVVKKF
171





403
QFPQGEAGYIAKSF
163





404
QFYWDQSNYGEQKP
161





405
HVPWAGSSDVLKVT
160





406
QVPWEKPVSFVQKS
159





407
HVWWEDPYYVVRKT
158





408
WLHWEQPYWTVKKS
143





409
LFAGENGYFTVRQF
139





410
QVSWVYPPNDVIVS
133





411
NDPWPVPYYMVKNS
130





412
QVQWQVPYYVVKKP
128





413
HFPLQEPYYVVKNS
127





414
QGSSVATSSVLTML
123





415
QPCADKPYCVHTIT
123





416
HVWWEEAHYSWKHS
120





417
DLFVEEPYLGVIKF
117





418
YSLWEDLIFNMQNS
116





419
QIPWVEASNVLISF
115





420
QFGQENHNHFVRKS
113





421
QVSLVESRSVLKIL
110





422
AVQRAKPYYAVONS
109





423
HLLGDYRFWAFTNA
109





424
ESPWAPPYYVVKKS
107





425
QIPTVEPYYEGKKS
105





426
QVRGGQPDQVLRLS
105





427
FVARNEPFYVVGTS
104





428
QIPLEEMYYVVRTY
103





429
QVSSVGPNHVVKKI
102





430
QLPWGKPYNVVVYS
101





431
QTPWQVPGYNMLNS
 99





432
AYLSEQLNYVMKNS
 98





433
QASSEAPSYPVVKS
 95





434
LDQWHTPNYVVRKS
 92





435
QLLWDPPSYFVNNS
 92





436
DVPWEQPYFVVRNS
 90





437
QVPYGQSFVVGTSS
 90





438
QLRWEDQYAHVEKS
 89





439
HVAWKAPYHFTKNS
 87





440
DGLYVRSFYVLQHS
 86





441
QFALKGLNGDFQKS
 83





442
QFPSAETYRIVKKF
 83





443
QLTWELGYHAFVKP
 83





444
QLLRNLSYDGAKYS
 82





445
QLRVKLQYDVIANN
 81





446
QVPAREPNHIVKKS
 80





447
SAREQGTFYFVNNS
 80





448
LFPKYAPHESVKMS
 78





449
QVPGNELYFVLKQY
 77


















TABLE 2G





SEQ ID




NO: #
Plated Nerve2
Count







450
QFAWSQSYAYSSTT
107





451
QGQWDKPYDAVQNS
 88





452
QIPGDAQYRLVKRS
 80





453
QIPWTQPQYDVTPP
 79





454
HVELESSYYFVNNS
 78





455
QFPLAEPYYHLTES
 77





456
HFDWEEPYYDASKA
 73





457
SYEWEDPYYDLKYS
 73





458
NVTGAEYYYRMNNY
 67





459
HVSCEVPCYGALAS
 66





460
HDASEGAYYVERNS
 65





461
QFGWEEPYYVLHTV
 65





462
HLSREGTYYAVKQP
 64





463
QLPLYDPYYTVRTS
 63





464
ELPYEEPGTFVEKA
 62





465
QFPWEEPYYVFTNS
 62





466
QIPGGERSYVLENS
 62





467
QITSDKPYYVVKNS
 60





468
YDPSGEPYYRVNNH
 59





469
QVPRPASSYIGKTS
 58





470
QIPLEGPTYVTNPS
 57





471
AIPSGAPYYKVNKY
 56





472
EFRSEQPYYFVNKY
 55





473
QIRWDETYYDIQKT
 55





474
QLALEQQSYDAKMS
 55





475
ALPWEHPWCVSKNC
 54





476
HVPVLEPYDLVKKS
 54





477
QIRSTEPYYVFKKT
 54





478
QSPGTNPQNVLIKS
 54





479
QITSVDPSSLAYRS
 53





480
QMPSGAPYDATLNS
 53





481
QNPWDEPSYVVKNV
 53





482
QSPFDETAYGLTHS
 53





483
QVLFAKSYYSVKDS
 53





484
RFPADEEHYVGKLS
 53





485
YVPYEEQKYVVTKS
 53





486
HSQGQTTEYVGKQS
 52





487
DFQSEETRNLAKMP
 51





488
DVPWPAPYYAVSKY
 51





489
QFPGEESHYVVRNS
 51





490
QFQWETPSHVINDS
 51





491
QTPRQTLYYSVKAY
 51





492
HVAYQEPFVYQKTS
 50





493
QISRGDTYYVMNKS
 50





494
HIPGEEPSDGVRSY
 49





495
HLSVAEPYYAVRNS
 49





496
QLQWQKSNWVGKIT
 49





497
EIPWEGTYRLVKHS
 48





498
HVPQKEPYYVVKTF
 48





499
QFPLFTTFYNVEES
 47





500
QFPWETSYYIVKNS
 47





501
QLAGGDPYVLPKLS
 47





502
DVASGEQSYIVVKY
 46





503
HVWLEDTISAEKKS
 46





504
QVQWDGASSIVQKT
 46





505
HFRGPEPYKFVENP
 45





506
HVPWEGPEYIVTQS
 45





507
QFPRHDPNEVVHKS
 45





508
QIPSEEPNYILKSS
 45





509
QRPGDEPSYFVKNS
 45





510
ELTMEQPSYVGNVS
 44





511
QLPWHEAFFGVNNG
 44





512
QVLEKEPSSNVHKS
 44





513
HVPSYAPNYEIKES
 43





514
QHLGEEPSYSGEYA
 43





515
SISSKDPYALVKYS
 43





516
QFPWDEPNYVVKNF
 42





517
QLPFNEPYYSQQVS
 42





518
HLSMEAPYYGVKEY
 41





519
HVLGEQPYSVVKTS
 41





520
HVSWEEPRYIVKDS
 41





521
QLPWEGPYYIVKKS
 41





522
QTPAEKPYYVVKKY
 41





523
HVPSQEWYYDLHKS
 40





524
QFPWHKQFYVGKSS
 40





525
DFQIVGPSYVGKNS
 39





526
HLPSEKPWYDLDGS
 39





527
HVSWSTSKYVVNHS
 39





528
QISWEDPDYGVRKS
 39





529
QVPCEQQSCVVKNS
 39





530
LFTGQQSYYEVQES
 38





531
LVSPTGPIYVMNKS
 38





532
QFASEEPYHVMKKP
 38





533
QFSRADSQYLVKNS
 38





534
QIPLQDPYHVVEKS
 38





535
QLLWGESYSIVKNY
 38





536
QLPSQTQNYYDTNA
 38





537
QLRSEDTYYTMQSY
 38





538
SVAGQQHYYVSTKP
 38





539
HLPCKKSFYACKKH
 37





540
HVRFAEPYYAGKTS
 37





541
LLTGAEPYYLGKRS
 37





542
QLPGAEAQNVVKYS
 37





543
QLRFDEHHSVVTHS
 37





544
QYPWEQRYYFERKS
 37





545
RVTWEGPYNSVKKS
 37





546
DVALGESHYNVKKY
 36





547
DVPNDLPSYLVKNS
 36





548
EFPYQESEYIVKQS
 36





549
GLPSKEPYYVVKPA
 36





550
HVPWDDQYYVWRTN
 36





551
HVQWDQTSYVVKNS
 36





552
HVTSDDSYYAVKNP
 36





553
QDPFGDLYYVTKMS
 36





554
QFPYDEPYNVVKKS
 36





555
QHPWKEQYYMATKP
 36





556
QLNSEEPYYHFKNY
 36





557
QLPTNDPYYVLMES
 36









Example 2: Prevalence of Amino Acid Residues in Nerve-Binding Peptides


FIG. 2 depicts the most prevalent amino acids at each position of the sample set of peptides identified in Example 1 (and listed in Table 2), based on unweighted (FIG. 2A) or weighted (FIG. 2B) probabilities. In an unweighted analysis, the probability of each amino acid at each position is based on its representation in the unique peptide set without considering the number of counts. In a weighted analysis, the probability of each amino acid in the sequence is based on its representation in the unique peptide set as a function of the total number of counts.


As shown in FIG. 2 and quantified in Table 3, by either analysis, the most frequent amino acid at each position corresponds to that in the original peptide sequences. However, some differences were noted with respect to less frequent amino acids at certain positions. For example, at position 10, the most frequently present amino acid was valine—the same as the parent peptide—in the unweighted and weighted alignments. However, the second (and third) most frequent amino acid at position 10 was leucine (and glycine) in the unweighted alignment but phenylalanine (and isoleucine) in the weighted analysis.









TABLE 3







Unweighted Probabilities





















position
1
2
3
4
5
6
7
8
9
10
11
12
13
14





AA
Q
V
P
W
E
E
P
Y
Y
V
V
K
K
S


percent
43.63
33.75
44.70
29.44
34.65
41.11
54.58
51.71
52.06
42.91
47.76
42.37
34.29
57.27



H
L
A
S
Q
Q
S
S
S
L
L
N
N
Y



27.11
17.24
9.52
18.85
11.85
10.77
11.49
10.59
8.44
9.52
12.75
12.03
18.13
8.44



D
F
S
G
D
D
T
N
H
A
G
T
T
P



6.10
15.80
8.62
10.95
11.31
8.80
6.82
7.18
7.90
7.90
8.08
10.05
7.18
7.90










Weighted Probabilities





















position
1
2
3
4
5
6
7
8
9
10
11
12
13
14





AA
Q
V
P
W
E
E
P
Y
Y
V
V
K
K
S


percent
43.28
33.73
44.92
31.25
36.67
41.89
56.36
54.41
50.13
45.57
47.09
46.03
36.10
55.05



H
L
A
S
Q
Q
S
S
N
F
L
N
N
T



27.12
16.43
9.38
17.52
13.42
12.36
10.65
8.55
10.02
7.76
10.61
11.50
17.67
9.19



D
F
T
G
D
D
L
N
S
I
G
R
Q
Y



7.09
15.74
8.76
9.67
9.50
8.71
6.41
5.99
8.25
7.50
10.17
10.18
6.79
8.88









Example 3: Alignments and Cluster Analysis of Nerve-Binding Peptides and

To further characterize the 557 distinct peptides and identify nerve-binding consensus sequences and corresponding formulas, clustering analyses were carried out using established platforms for multiple sequence alignments. See, e.g., Sievers and Higgins, 2014, Clustal Omega, accurate alignment of very large numbers of sequences, Methods Mol. Biol. 1079, 105-116; Sievers and Higgins, 2018, Clustal Omega for making accurate alignments of many protein sequences, Protein Sci. 27, 135-145; Sievers and Higgins, 2021, The Clustal Omega multiple alignment package, Methods Mol. Biol. 2231, 3-16; Larsson, 2014, AliView: a fast and lightweight alignment viewer and editor for large data sets, Bioinformatics. 30, 3276-3278; Waterhouse et al., 2009, Jalview Version 2—a multiple sequence alignment editor and analysis workbench, Bioinformatics. 25, 1189-1191; Procter et al. 2021, Alignment of biological Sequences with Jalview, Methods Mol Biol. 2231, 203-224.


In the first approach, Clustal Omega was used to create an initial phylogenetic tree, and 14 highly related clusters were chosen to visualize individually. The 14 clusters were each visualized with AliView software and the “majority rule consensus” residues at each position were highlighted in order to identify patterns in each cluster. These 14 clusters correspond to Formulas I to XIV (and Subformulas Ia to XIVa), respectively, and are depicted in FIGS. 3 to 16, respectively. Table 4 shows the “majority rule” consensus sequence for these 14 clusters, which are each based on different, non-overlapping sets of peptides in the representative sample.
























TABLE 4





Position
1
2
3
4
5
6
7
8
9
10
11
12
13
14








Parent
Q
V
P
W
E
E
P
Y
Y
V
V
K
K
S
(SEQ ID


Peptide














NO: 558)





Cluster 1
H
V
—
—
—
—
P
Y
Y
—
V
—
—
S






Cluster 2
—
—
—
—
E
—
—
Y
Y
—
V
—
—
—






Cluster 3
H
V
P
—
—
G
P
Y
—
V
—
K
—
—






Cluster 4
H
V
—
—
—
—
—
—
Y
V
—
K
—
—






Cluster 5
H
V
—
W
—
E
P
Y
Y
V
—
—
K
—






Cluster 6
—
—
—
—
—
E
—
—
—
—
—
—
—
S






Cluster 7
Q
—
—
W
E
—
P
—
Y
V
—
K
—
S






Cluster 8
Q
—
P
—
—
—
P
—
Y
—
V
—
—
—






Cluster 9
Q
—
P
W
E
—
P
Y
—
—
—
—
—
S






Cluster 10
Q
—
—
—
—
E
P
Y
Y
—
—
K
—
—






Cluster 11
Q
—
P
—
—
—
—
—
Y
—
—
K
—
S






Cluster 12
Q
—
P
—
—
E
P
Y
—
—
—
—
—
S






Cluster 13
H
—
A
—
—
E
P
—
—
—
V
—
—
S






Cluster 14
H
V
—
S
—
E
P
Y
Y
—
L
—
—
S









In the second approach, an alternate relational tree was created with Jalview software, and 30 smaller clusters (clusters 15 through 44) were chosen. These 30 clusters were each visualized with AliView software and the “majority rule consensus” residues at each position were highlighted in order to identify patterns in each cluster.


These 30 clusters correspond to Formulas XV to XLIV (and Subformulas XVa to XLIVa), respectively, and are depicted in FIGS. 17 to 46, respectively. Table 5 shows the “majority rule” consensus sequence for these clusters (numbered 15 to 44), which are each based on different, non-overlapping sets of peptides in the representative sample.
























TABLE 5





Position
1
2
3
4
5
6
7
8
9
10
11
12
13
14








Parent
Q
V
P
W
E
E
P
Y
Y
V
V
K
K
S
(SEQ ID


Peptide














NO: 558)





Cluster 15
—
—
—
—
—
—
P
Y
—
—
—
T
Q
T






Cluster 16
Q
F
P
—
—
E
P
Y
Y
—
—
K
K
T






Cluster 17
Q
S
P
—
D
E
—
Y
Y
G
V
—
—
A






Cluster 18
Q
L
P
L
—
—
—
N
Y
—
V
T
—
S






Cluster 19
Q
—
P
—
E
—
P
—
Y
—
V
—
K
—






Cluster 20
Q
L
P
G
E
—
—
Y
—
L
—
K
K
S






Cluster 21
Q
—
P
S
—
E
P
Y
—
—
—
K
—
S






Cluster 22
Q
—
P
—
—
—
P
—
S
—
V
—
—
—
—





Cluster 23
Q
L
P
W
—
—
P
Y
—
V
V
—
—
S






Cluster 24
Q
—
P
—
—
—
P
Y
Y
V
—
—
—
S






Cluster 25
Q
—
P
—
—
—
P
Y
—
V
V
—
K
S






Cluster 26
Q
—
—
—
—
—
Q
Y
—
V
—
N
—
S






Cluster 27
Q
F
P
—
E
—
S
—
Y
—
V
K
—
S






Cluster 28
Q
—
—
—
—
—
S
—
—
—
—
—
K
S






Cluster 29
D
V
—
—
—
—
—
—
Y
—
—
—
—
—






Cluster 30
H
—
P
—
—
E
—
—
Y
V
—
K
—
S






Cluster 31
—
V
S
—
—
E
P
—
—
V
L
—
—
—






Cluster 32
Q
—
—
—
E
E
P
Y
Y
V
—
K
—
—






Cluster 33
Q
—
—
W
E
—
P
—
Y
—
—
—
K
S






Cluster 34
—
—
P
—
—
—
P
—
Y
—
V
—
N
S






Cluster 35
—
—
—
—
E
—
P
Y
Y
—
V
—
—
S






Cluster 36
H
V
—
—
—
E
P
Y
—
—
V
—
K
S






Cluster 37
H
V
—
—
—
—
—
Y
Y
—
—
K
N
S






Cluster 38
H
L
P
—
E
E
—
Y
—
—
V
—
—
—






Cluster 39
Q
—
—
W
—
—
P
Y
Y
—
V
K
K
—






Cluster 40
—
—
—
W
—
E
—
—
—
V
V
K
N
—






Cluster 41
Q
—
—
—
—
E
—
—
—
V
V
—
K
S






Cluster 42
H
V
P
—
—
G
—
Y
—
V
—
—
—
—






Cluster 43
H
V
P
—
Q
E
P
Y
Y
V
V
K
—
—






Cluster 44
Q
—
—
—
—
—
P
Y
—
—
V
K
—
S









The various embodiments described above can be combined to provide further embodiments. All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and/or listed in the Application Data Sheet are incorporated herein by reference, in their entirety. Aspects of the embodiments can be modified, if necessary, to employ concepts of the various patents, applications and publications to provide yet further embodiments. Moreover, in certain instances, well-known methods, procedures, or other specific details have not been described to avoid unnecessarily obscuring aspects of the invention defined by the appended claims


These and other changes can be made to the embodiments in light of the above-detailed description. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the disclosure.

Claims
  • 1. A nerve targeting peptide conjugate comprising: (a) a peptide comprising the amino acid sequence of any one of SEQ ID NOS:1-557; and(b) a cargo molecule.
  • 2.-8. (canceled)
  • 9. A nerve targeting peptide conjugate comprising: (a) a peptide comprising the amino acid sequence of Formula (I): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, E, H, L, Q, R, or Y;X2 is D, F, G, I, L, N, or V;X3 is A, E, H, I, L, P, Q, R, S, T, or W;X4 is C, F, G, K, L, Q, R, S, V, W, or Y;X5 is A, D, E, G, K, L, Q, T, V, or Y;X6 is A, D, E, G, K, N, Q, R, S, V, or Y;X7 is A, L, M, P, S, T, or W;X8 is C, E, H, L, N, P, R, S, V, or Y;X9 is A, C, D, E, H, L, N, P, S, or Y;X10 is A, D, E, F, G, H, I, L, N, P, S, T, V, or Y;X11 is A, D, E, I, L, Q, R, T, or V;X12 is D, I, K, L, N, Q, R, S, or T;X13 is A, C, D, E, H, I, K, M, N, Q, R, T, or V; andX14 is P, S, T, V, or Y;and(b) a cargo molecule.
  • 10. (canceled)
  • 11. (canceled)
  • 12. A nerve targeting peptide conjugate comprising: (a) a peptide comprising the amino acid sequence of Formula (II): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein:X1 is A, D, E, H, K, L, N, Q, R, S, or T;X2 is A, F, H, I, L, T, or V;X3 is A, E, F, L, P, Q, S, or T;X4 is A, E, F, G, L, M, R, S, T, V, W, or Y;X5 is A, E, G, H, I, P, Q, T, V, or Y;X6 is A, D, E, G, H, I, K, N, Q, or R;X7 is A, H, L, P, Q, R, S, or T;X8 is G, H, L, R, S, V, or Y;X9 is D, E, F, L, M, N, T, V, or Y;X10 is A, D, F, G, H, I, L, N, R, S, V, or Y;X11 is E, F, G, K, L, M, R, or V;X12 is E, H, I, K, N, P, Q, R, S, T, or V,X13 is D, E, F, H, I, K, N, Q, R, S, T, or V; andX14 is A, F, H, P, R, S, or T; and(b) a cargo molecule.
  • 13. (canceled)
  • 14. (canceled)
  • 15. A nerve targeting peptide conjugate comprising: (a) a peptide comprising an amino acid sequence of Formula (III): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is H, Q, R, or V;X2 is F, G, H, I, L, or V;X3 is A, H, P, or T;X4 is F, G, L, M, S, T, or W;X5 is A, D, G, I, K, P, Q, or V;X6 is D, E, G, or P;X7 is L, P, Q, R, S, or T;X8 is F, H, S, or Y;X9 is D, F, G, H, N, S, or Y;X10 is N or V;X11 is A, E, G, K, L, Q, S, V, or W;X12 is A, D, E, G, K, M, N, Q, R, or T;X13 is E, H, K, N, Q, R, T, or V; andX14 is F, H, N, P, S, T, or Y; and(b) a cargo molecule.
  • 16. (canceled)
  • 17. (canceled)
  • 18. A nerve targeting peptide conjugate comprising: (a) a peptide comprising the amino acid sequence of Formula (IV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is E, H, K, L, Q, R, S, V, or Y;X2 is A, D, F, G, H, I, L, S, V, or Y;X3 is A, H, K, L, N, P, Q, R, S, T, V, or W;X4 is A, C, E, F, G, I, L, M, P, R, S, T, V, W, or Y;X5 is A, D, E, G, H, I, K, N, Q, S, T, V, or Y;X6 is A, D, E, G, H, K, N, P, Q, S, T, V, or Y;X7 is A, H, L, P, Q, R, S, or T;X8 is C, D, E, F, H, I, K, L, N, S, V, or Y;X9 is A, D, F, H, K, L, N, S, V, W, or Y;X10 is A, E, F, G, I, L, P, R, S, V, or Y;X11 is A, C, E, F, G, I, L, M, N, P, Q, S, T, V, or W;X12 is A, E, H, I, K, N, P, Q, R, T, V, or W;X13 is D, E, H, I, K, M, N, Q, R, S, T, or Y; andX14 is A, F, H, I, M, P, R, S, T, W, or Y; and(b) a cargo molecule.
  • 19. (canceled)
  • 20. (canceled)
  • 21. A nerve targeting peptide conjugate comprising: (a) a peptide comprising the amino acid sequence of Formula (V): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, F, G, H, N, Q, R, T, or Y;X2 is D, F, H, I, L, M, Q, or V;X3 is D, E, G, L, P, Q, R, S, or T;X4 is G, L, Q, R, S, or W;X5 is A, D, E, G, H, K, Q, R, or V;X6 is A, E, G, Q, or V;X7 P or R;X8 is S or Y;X9 is G, H, N, Q, R, S, T, or Y;X10 is D, F, I, P, or V;X11 is A, D, F, G, I, L, Q, or V;X12 is A, E, H, K, L, N, R, S, or T;X13 is K, M, N, P, Q, T, or W; andX14 is A, F, I, L, P, R, S, or Y;and(b) a cargo molecule.
  • 22. (canceled)
  • 23. (canceled)
  • 24. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (VI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, H, L, N, Q, or Y;X2 is A, D, F, G, H, I, L, R, or V;X3 is A, F, L, P, S, T, or V;X4 is A, G, L, S, V, W, or Y;X5 is A, D, E, I, K, Q, or V;X6 is A, E, L, N, P, Q, or R;X7 is A, G, H, L, P, Q, R, or S;X8 A, D, F, K, N, R, S, or Y;X9 is D, E, F, K, L, N, S, or Y;X10 is A, E, F, G, I, L, N, P, T, or V;X11 is E, K, L, M, V, or W;X12 is E, H, K, L, Q, R, S, or T;X13 is E, H, K, M, N, Q, or Y; andX14 is F, G, I, S, or T;and(b) a cargo molecule.
  • 25. (canceled)
  • 26. (canceled)
  • 27. A nerve targeting peptide conjugate, comprising: (a) a peptide comprises or consists of the amino acid sequence of Formula (VII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, D, E, H, K, L, N, Q, R, or Y;X2 is A, C, D, E, F, G, I, L, R, S, or V;X3 is A, E, H, L, P, Q, R, S, T, W, or Y;X4 is C, F, G, I, L, M, S, T, V, or W;X5 is D, E, G, H, Q, S, or V;X6 is A, D, E, G, H, Q, T, V, or Y;X7 is H, L, P, Q, R, S, T, or V;X8 is C, D, E, F, H, L, N, Q, S, W, or Y;X9 is C, D, G, H, N, P, Q, S, W, or Y;X10 is A, D, G, I, L, P, T, or V;X11 is A, C, F, G, I, K, L, N, R, S, T, or V;X12 is D, K, N, Q, R, T, V, or Y;X13 is D, E, I, K, L, M, N, P, Q, R, T, or V; andX14 is C, F, S, V, or Y;and(b) a cargo molecule.
  • 28. (canceled)
  • 29. (canceled)
  • 30. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (VIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, D, E, H, L, N, Q, R, S, T, Y;X2 is A, D, E, F, H, I, L, N, P, R, S, T, V, Y;X3 is C, I, L, P, Q, R, S, T, V;X4 is A, C, E, F, G, K, L, M, N, Q, R, S, T, V, W, Y;X5 is A, D, E, G, H, K, P, Q, R, S, T, V, Y;X6 is A, D, E, G, H, K, L, N, P, Q, R, S, T, V, W, Y;X7 is A, D, F, H, I, L, M, P, Q, S, T, W;X8 is C, F, G, H, I, K, L, N, P, Q, R, S, T, V, W, Y;X9 is A, C, D, E, F, G, H, N, S, T, Y;X10 is A, D, F, G, I, K, L, M, N, P, R, S, V;X11 is A, E, G, H, I, M, P, Q, V, Y;X12 is A, E, H, I, K, L, N, P, Q, R, T, V;X13 is A, D, E, I, K, M, N, P, R, S, T, V, Y; andX14 is A, C, F, G, H, L, N, P, S, T, Y;and(b) a cargo molecule.
  • 31. (canceled)
  • 32. (canceled)
  • 33. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (IX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, H, N, Q, R, S, W, or Y;X2 is D, F, H, I, L, M, N, P, R, S, V, or Y;X3 is A, E, H, L, P, Q, R, S, T, or Y;X4 is A, G, H, I, L, N, P, R, S, or W;X5 is A, E, G, I, K, Q, S, or V;X6 is A, D, E, G, K, L, P, Q, R, T, or V;X7 is A, G, K, L, P, Q, R, S, or T;X8 is E, I, K, L, S, V, or Y;X9 is A, D, F, G, H, I, K, N, Q, S, T, W, or Y;X10 is A, D, F, H, I, L, N, T, or V;X11 is A, D, E, F, G, L, M, V, or W;X12 is E, K, N, Q, R, T, or V;X13 is E, H, K, L, N, Q, R, S, T, or Y; andX14 is E, P, R, S, T, or Y;and(b) a cargo molecule.
  • 34. (canceled)
  • 35. (canceled)
  • 36. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (X): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, D, E, G, H, K, N, Q, R, S, T, or Y;X2 is A, D, F, H, I, L, M, Q, S, T, V, or Y;X3 is A, E, G, K, L, N, P, Q, R, S, T, V, or Y;X4 is C, F, G, H, L, M, R, S, T, W, or Y;X5 is A, D, E, G, K, L, N, Q, T, or V;X6 is A, D, E, G, H, K, L, N, Q, S, or V,X7 is A, L, P, R, S, T, or Y;X8 is A, C, D, F, G, N, R, S, T, or Y;X9 is A, D, F, H, L, N, Q, R, S, T, or Y;X10 is A, D, E, F, G, H, I, L, P, R, S, T, or V;X11 is A, D, E, F, G, H, I, L, M, P, Q, R, S, or V;X12 is A, D, E, G, H, K, N, Q, S, T, or V;X13 is D, E, F, H, I, K, L, M, N, Q, R, S, T, V, or Y; andX14 is A, F, H, I, K, L, N, P, R, S, T, V, or Y;and(b) a cargo molecule.
  • 37. (canceled)
  • 38. (canceled)
  • 39. A nerve targeting peptide conjugate of Formula (XI), comprising: (a) a peptide comprising the amino acid sequence of Formula (XI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, H, L, Q, or R;X2 is F, G, H, I, L, N, Q, S, T, V, or Y;X3 is A, G, H, L, P, Q, R, S, T, or Y;X4 is A, C, F, G, L, P, Q, R, S, W, or Y;X5 is A, D, E, G, H, K, N, P, Q, S, T, V, or Y;X6 is A, D, E, G, H, K, L, N, Q, T, or V;X7 is A, E, H, K, L, P, Q, R, S, or T;X8 is C, D, E, F, G, H, K, L, N, P, Q, S, T, V, or Y;X9 is A, D, F, G, H, I, L, N, Q, S, T, V, W, or Y;X10 is A, D, F, G, H, I, L, P, R, S, T, V, or Y;X11 is A, D, E, F, G, H, L, M, Q, S, or V;X12 is A, E, H, I, K, L, N, P, Q, R, S, or T;X13 is A, D, H, I, K, L, M, N, P, Q, S, T, V, or Y; andX14 is A, F, H, L, P, S, T, V, or Y;and(b) a cargo molecule.
  • 40. (canceled)
  • 41. (canceled)
  • 42. A nerve targeting peptide conjugate, comprising: (a) the amino acid sequence of Formula (XII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is E, H, K, L, N, Q, or R;X2 is A, C, D, F, I, K, L, M, N, S, V, or Y;X3 is A, L, P, R, S, or T;X4 is A, E, F, G, L, M, R, S, T, W, or Y;X5 is A, D, E, F, G, H, K, N, Q, R, T, V, or Y;X6 is A, D, E, G, Q, R, S, T, V, or Y;X7 is A, H, K, L, P, Q, R, S, or T;X8 is A, C, D, F, H, N, R, S, T, or Y;X9 is D, E, F, H, L, N, R, S, T, V, or Y;X10 is A, D, F, G, H, I, L, N, R, S, T, or V;X11 is A, D, F, G, L, M, P, Q, R, T, V, or W;X12 is A, D, E, H, K, L, M, N, P, Q, R, S, T, or Y;X13 is E, H, I, K, L, M, N, P, Q, R, S, T, or V; andX14 is A, F, P, or S;and(b) a cargo molecule.
  • 43. (canceled)
  • 44. (canceled)
  • 45. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XIII): X1-X2-X3-X4-X5-E-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is E, F, H, or Q;X2 is A, F, I, M, S, T, or V;X3 is A, G, H, P, Q, R, or T;X4 is F, G, L, M, N, R, S, W, or Y;X5 is A, G, K, N, P, Q, or R;X7 is H, P, R, or T;X8 is D, F, H, K, L, N, or Y;X9 is F, G, H, K, Q, T, V, or Y;X10 is F, G, L, P, S, V, or Y;X11 is L, M, Q, or V;X12 is E, G, I, K, L, N, P, S, or Y;X13 is E, H, I, N, Q, S, or T; andX14 is I, P, S, or Y;and(b) a cargo molecule.
  • 46. (canceled)
  • 47. (canceled)
  • 48. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XIV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is H, L, N, Q, or R;X2 is F, L, R, S, or V;X3 is P, S, or T;X4 is A, L, S, or T;X5 is A, D, E, H, Q, R, V, or Y;X6 is D, E, or K;X7 is A, P, Q, or W;X8 is H, R, W, or Y;X9 is H, K, L, S, or Y;X10 is D, E, F, L, T, or V;X11 is A, D, or L;X12 is D, E, G, H, K, N, Q, or R;X13 is G, K, N, Q, or T; andX14 is P or S;and(b) a cargo molecule.
  • 49. (canceled)
  • 50. (canceled)
  • 51. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XV): X1-X2-X3-X4-X5-X6-X7-Y-X9-X10-X11-X12-X13-X14, wherein: X1 is E, K, Q, or T;X2 is D, F, L, P, or V;X3 is C, L, P, T, or V;X4 is A, L, or T;X5 is D, E, H, or Q;X6 is A, D, K, or N;X7 is A, P, or S;X9 is C, H, M, or Y;X10 is F, L, or V;X11 is E, G, H, R, or V;X12 is K or T;X13 is I or Q; andX14 is A, F, or T;and(b) a cargo molecule.
  • 52. (canceled)
  • 53. (canceled)
  • 54. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XVI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, H, K, or Q;X2 is F, I, or V;X3 is A, P, R, S, or T;X4 is G, L, M, Q, S, T, or W;X5 is D, G, Q, T, or V;X6 is D, E, H, Q, or S;X7 is A, P, R, or T;X8 is E, G, T, V, or Y;X9 is A, Q, S, or Y;X10 is A, D, E, G, I, L, R, T, V, or Y;X11 is A, E, F, G, or I;X12 is K, Q, or T;X13 is E, I, K, M, N, or S; andX14 is F, L, S, or T;and;(b) a cargo molecule.
  • 55. (canceled)
  • 56. (canceled)
  • 57. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XVII): Q-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X2 is H, S, or T;X3 is P, Q, or T;X4 is F, M, P, or W;X5 is D, E, or Q;X6 is E or P;X7 is P, R, or T;X8 is A, K, or Y;X9 is H or Y;X10 is G, R, or V;X11 is L or V;X12 is I, L, S, or T;X13 is H, K, M, or Q; andX14 is A or S;and(b) a cargo molecule.
  • 58. (canceled)
  • 59. (canceled)
  • 60. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising an amino acid sequence of Formula (XVIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is K, Q, or R;X2 is F or L;X3 is G, P, or T;X4 is E, L, Q, S, or W;X5 is A, D, E, F, P, Q, V, or Y;X6 is A, D, E, K, N, Q, T, or Y;X7 is H, L, P, Q, R, or T;X8 is F, N, or Y;X9 is H, T, or Y;X10 is F, H, I, L, N, T, or Y;X11 is D, G, L, M, Q, or V;X12 is D, E, K, R, or T;X13 is A, E, K, M, N, P, Q, R, or T; andX14 is A, S, or P;and(b) a cargo molecule.
  • 61. (canceled)
  • 62. (canceled)
  • 63. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XIX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is Q or T;X2 is H, I, L, T, or V;X3 is P or T;X4 is A, C, F, L, R, S, T, or Y;X5 is A, E, G, H, or Q;X6 is A, E, K, Q, T, V, or Y;X7 is L, M, P, Q, or S;X8 is C, F, L, N, W, or Y;X9 is C or Y;X10 is A, L, P, S, or V;X11 is A, G, or V;X12 is K, N, Q, R, or V;X13 is A, E, K, T, V, or Y; andX14 is F, G, N, P, S, T, or Y.
  • 64. (canceled)
  • 65. (canceled)
  • 66. A nerve targeting peptide conjugate, comprising: (a) the amino acid sequence of Formula (XX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is E, H, N, Q, or S;X2 is H, I, L, or V;X3 is A, P, or T;X4 is G, L, M, or W;X5 is D, E, G, or V;X6 is A, D, E, G, H, L, or V;X7 is G, K, P, Q, R, or T;X8 is E, L, S, or Y;X9 is H, K, N, Q, R, S, V, or Y;X10 is A, F, I, L, or P;X11 is F, G, I, L, M, P, R, or V;X12 is H, K, N, or V;X13 is H, K, L, M, R, S, or T; andX14 is F, P, R, S, or Y;and(b) a cargo molecule.
  • 67. (canceled)
  • 68. (canceled)
  • 69. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXI): X1-X2-X3-X4-X5-E-X7-X8-X9-X10-X11-K-X13-X14, wherein: X1 is D, Q, or R;X2 is A, F, H, I, L, V, or Y;X3 is A, E, L, P, or Q;X4 is A, L, R, S, or V;X5 is A, E, G, H, K, Q, or R;X7 is K, L, P, R, or T;X8 is D, N, R, or Y;X9 is E, F, H, N, R, S, or T;X10 is A, D, F, I, L, R, or V;X11 is A, F, L, M, P, Q, V, or W;X13 is E, H, K, M, or R; andX14 is P, F, or S;and(b) a cargo molecule.
  • 70. (canceled)
  • 71. (canceled)
  • 72. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, H, L, or Q;X2 is D, F, I, L, N, S, T, or Y;X3 is P, Q, S, or T;X4 is A, C, L, S, or W;X5 is A, D, E, H, K, Q, S, T, or V;X6 is A, D, E, G, H, K, Q, R, S, W, or Y;X7 is D, P, or S;X8 is C, F, G, H, L, N, Q, S, T, W, or Y;X9 is A, C, D, F, N, S, or Y;X10 is A, D, G, L, or V;X11 is A, G, L, Q, S, T, V, W, or Y;X12 is A, D, K, P, Q, S, T, or Y;X13 is I, K, M, N, P, R, or T; andX14 is A, C, F, P, S, or Y;and(b) a cargo molecule.
  • 73. (canceled)
  • 74. (canceled)
  • 75. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, or Q;X2 is L, M, N, P, R, or V;X3 is P, Q, R, or T;X4 is R, S, or W;X5 is E, G, H, Q, T, or Y;X6 is A, E, K, N, Q, or V;X7 is A, P, R, or S;X8 is D, F, H, K, N, or Y;X9 is D, F, H, N, S, T, W, or Y;X10 is G, I, R, or V;X11 is D, G, M, or V;X12 is E, K, N, R, or V;X13 is I, K, N, Q, R, S, T, V, or Y; andX14 is G, H, P, S, T, or Y;and(b) a cargo molecule.
  • 76. (canceled)
  • 77. (canceled)
  • 78. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXIV): Q-X2-P-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-S wherein: X2 is D, I, K, L, or M;X4 is F, G, L, S, or T;X5 is E, G, H, N, Q, V, or Y;X6 is A, D, E, G, or S;X7 is L, P, or S;X8 is N, T, or Y;X9 is D, N, or Y;X10 is A, G, I, S, or V;X11 is D, G, L, Q, or T;X12 is K, L, M, N, Q, R, S, or T; andX13 is E, K, M, N, P, S, or V;and(b) a cargo molecule.
  • 79. (canceled)
  • 80. (canceled)
  • 81. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXV): Q-X2-P-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X2 is F, I, L, N, S, or V;X4 is E, G, L, R, W, or Y;X5 is A, D, G, H, K, N, Q, S, T, V, or Y;X6 is A, D, E, H, N, Q, S, or T;X7 is A, K, L, P, or R;X8 is N, Q, S, T, or Y;X9 is E, F, H, N, V, or Y;X10 is S or V;X11 is F, H, L, M, S, or V;X12 is E, H, I, K, or P;X13 is K, N, Q, S, or Y; andX14 is F, S, V, or Y; and(b) a cargo molecule.
  • 82. (canceled)
  • 83. (canceled)
  • 84. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXVI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is N, Q, R, S, or V;X2 is A, D, F, H, I, L, N, or V;X3 is L, P, Q, R, or S;X4 is A, C, E, G, H, I, L, R, S, V, W, or Y;X5 is E, G, K, N, P, Q, or S;X6 is A, D, E, G, K, L, or R;X7 is H, L, Q, S, or T;X8 is F, H, K, or Y;X9 is D, F, I, L, N, or Y;X10 is A, F, G, L, M, or V;X11 is A, F, G, I, L, M, P, V, or W;X12 is A, K, N, T, or Y;X13 is H, K, L, M, N, Q, S, T, or Y; andX14 is C, E, G, N, P, or S;and(b) a cargo molecule.
  • 85. (canceled)
  • 86. (canceled)
  • 87. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXVII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, Q, or R;X2 is F, G, I, L, or V;X3 is A, H, P, Q, R, or S;X4 is C, L, R, S, T, W, or Y;X5 is A, E, G, H, P, Q, or V;X6 is A, D, E, G, K, Q, or T;X7 is A, H, Q, R, or S;X8 is C, E, F, N, Q, S, or Y;X9 is C, H, S, V, W, or Y;X10 is D, I, L, T, or V;X11 is A, E, G, or V;X12 is K, P, or T;X13 is K, M, N, Q, R, S, or T; andX14 is S or Y.
  • 88. (canceled)
  • 89. (canceled)
  • 90. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXVIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is H, Q, or S;X2 is C, F, G, I, V, or Y;X3 is A, P, Q, R, S, T, or Y;X4 is C, L, N, S, W, or Y;X5 is D, E, H, K, Q, S, or V;X6 is A, E, G, K, or Q;X7 is L or S;X8 is C, N, P, S, V, or Y;X9 is A, D, F, G, N, S, or Y;X10 is D, G, L, S, V, or Y;X11 is A, E, F, G, L, Q, S, or V;X12 is A, K, N, Q, R, S, or T;X13 is I, K, or T; andX14 is I, L, P, R, S, T, or W;and(b) a cargo molecule.
  • 91. (canceled)
  • 92. (canceled)
  • 93. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXIX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D or E;X2 is F, G, H, I, R, or V;X3 is A, L, P, or T;X4 is F, G, L, R, S, T, W, or Y;X5 is A, D, E, G, I, P, R, or V;X6 is A, D, E, H, K, L, or R;X7 is L, P, Q, R, S, T, or W;X8 is F, H, N, R, S, V, or Y;X9 is F, K, or Y;X10 is A, D, F, G, I, L, N, R, or V;X11 is A, E, F, G, L, M, or V;X12 is E, K, P, Q, S, T, or V;X13 is D, F, H, I, K, or Q; andX14 is G, P, S, T, or Y;and(b) a cargo molecule.
  • 94. (canceled)
  • 95. (canceled)
  • 96. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, H, L, or R;X2 is F, G, H, I, L, S, or V;X3 is L, P, or Q;X4 is A, C, G, L, P, Q, R, S, T, V, or W;X5 is D, E, G, H, K, or Q;X6 is D, E, K, T, V, or Y;X7 is A, E, L, P, R, S, T, or W;X8 is D, E, F, H, S, W, or Y;X9 is I, W, or Y;X10 is A, D, I, P, or V;X11 is A, C, F, G, H, I, L, M, T, or V;X12 is D, E, H, K, T, or V;X13 is G, H, I, K, L, N, Q, R, S, V, or Y; andX14 is A, F, H, I, M, S, or Y.
  • 97. (canceled)
  • 98. (canceled)
  • 99. A nerve targeting peptide conjugate, comprising: (a) a peptide comprises or consists of the amino acid sequence of Formula (XXXI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, H, L, Q, R, T, or Y;X2 is F, G, I, R, S, or V;X3 is A, G, K, L, P, R, S, or T;X4 is A, G, L, R, S, T, W, or Y;X5 is A, E, G, H, K, N, P, Q, T, V, or Y;X6 is A, D, E, G, or N;X7 is A, P, Q, S, or T;X8 is D, F, G, H, N, R, S, or Y;X9 is D, F, G, H, K, N, Q, R, S, T, V, or Y;X10 is E, I, L, N, T, V, or Y;X11 is A, F, G, K, L, M, Q, or V;X12 is I, K, L, N, Q, R, or T;X13 is E, H, I, K, M, N, Q, T, or W; andX14 is F, H, I, L, N, P, R, or S;and(b) a cargo molecule.
  • 100. (canceled)
  • 101. (canceled)
  • 102. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, H, L, N, Q, or Y;X2 is C, D, F, H, I, L, R, S, T, V, or Y;X3 is A, G, L, N, P, Q, R, T, V, or Y;X4 is C, F, G, R, S, or W;X5 is A, E, or T;X6 is A, D, E, Q, S, or Y;X7 is A, L, P, T, or Y;X8 is C, E, F, G, N, Q, S, or Y;X9 is H, S, or Y;X10 is A, F, H, L, R, S, T, or V;X11 is A, C, D, E, F, G, H, L, M, N, Q, or V;X12 is A, D, E, G, H, K, N, Q, S, or T;X13 is D, E, F, I, K, L, M, N, P, Q, R, S, T, or Y; andX14 is A, F, K, L, P, S, V, or Y;and(b) a cargo molecule.
  • 103. (canceled)
  • 104. (canceled)
  • 105. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, H, L, N, Q, R, or Y;X2 is A, D, E, F, I, L, T, V, or Y;X3 is A, E, L, P, Q, R, or S;X4 is F, G, P, R, S, or W;X5 is A, D, E, G, H, Q, S, or V;X6 is A, D, E, G, H, Q, or T;X7 is P, Q, R, S, or T;X8 is D, F, H, K, N, S, or Y;X9 is D, F, G, H, N, P, Q, S, or Y;X10 is A, D, F, G, I, L, P, R, or V;X11 is A, E, G, I, L, R, or V;X12 is D, E, K, N, Q, R, T, or V;X13 is D, E, I, K, L, Q, R, or T; andX14 is A, I, S, T, or Y;and(b) a cargo molecule.
  • 106. (canceled)
  • 107. (canceled)
  • 108. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXIV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, D, E, K, L, N, Q, R, S, T, or Y;X2 is D, E, F, I, L, N, Q, R, S, T, or V;X3 is I, L, P, Q, or T;X4 is F, G, I, K, M, N, Q, R, S, V, W, or Y;X5 is A, D, E, G, P, Q, V, or Y;X6 is A, D, E, G, H, L, P, Q, S, or V;X7 is F or P;X8 is D, G, K, N, R, S, V, or Y;X9 is D, E, L, or Y;X10 is D, F, G, K, L, M, N, R, or V;X11 is A, E, F, G, M, P, Q, R, or V;X12 is K, L, N, or V;X13 is A, D, K, M, N, T, or V; andX14 is A, F, H, L, P, S, T, V, or Y; and(b) a cargo molecule.
  • 109. (canceled)
  • 110. (canceled)
  • 111. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXV): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, E, H, L, N, Q, R, S, V, or Y;X2 is F, I, N, S, V, or Y;X3 is A, E, I, L, P, R, S, T, V, W, or Y;X4 is C, F, G, K, L, R, S, W, or Y;X5 is D, E, I, K, Q, V, or Y;X6 is A, D, E, G, N, P, Q, R, S, V, or Y;X7 is G, L, P, Q, or S;X8 is H, I, L, N, S, or Y;X9 is A, D, E, F, L, N, S, T, V, or Y;X10 is A, D, E, F, G, H, I, L, N, S, T, or V;X11 is K, L, M, or V;X12 is I, K, N, P, Q, R, S, or T;X13 is A, C, E, H, K, M, N, Q, R, S, T, or Y; andX14 is F, H, P, S, T, or V; and(b) a cargo molecule.
  • 112. (canceled)
  • 113. (canceled)
  • 114. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXVI): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, F, H, L, N, Q, R, S, T, or Y;X2 is A, D, F, G, I, L, Q, or V;X3 is A, D, E, F, G, H, K, L, P, Q, R, S, T, or V;X4 is A, C, E, F, G, I, L, P, Q, R, S, V, W, or Y;X5 is A, D, E, G, H, I, K, L, N, P, Q, R, S, T, V, or Y;X6 is A, D, E, G, K, N, P, Q, S, T, V, or Y;X7 is A, H, I, L, P, Q, R, S, or T;X8 is A, E, F, H, I, K, L, P, Q, R, S, V, or Y;X9 is A, C, D, F, G, H, K, L, N, P, Q, R, S, or Y;X10 is A, D, E, F, G, H, I, L, N, P, S, V, or Y;X11 is A, D, E, F, G, I, L, M, N, S, T, or V;X12 is D, E, G, H, I, K, N, Q, R, S, T, W, or Y;X13 is D, E, H, K, N, Q, R, S, T, or V; andX14 is A, I, P, S, or Y.
  • 115. (canceled)
  • 116. (canceled)
  • 117. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXVII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, E, H, Q, or R;X2 is D, F, G, L, M, or V;X3 is A, E, H, P, Q, R, S, T, or W;X4 is C, G, L, R, S, V, or W;X5 is A, D, E, G, I, K, L, or Y;X6 is A, D, E, G, K, L, N, Q, S, or V;X7 is A, H, L, M, P, S, T, or W;X8 is C, H, I, L, N, S, V, or Y;X9 is D, H, L, N, S, or Y;X10 is A, D, E, F, G, L, P, S, T, or V;X11 is A, D, E, G, I, M, Q, R, T, V, or W;X12 is H, I, K, L, N, Q, R, or T;X13 is A, E, H, I, K, M, N, R, or V; andX14 is F, P, S, or T;and(b) a cargo molecule.
  • 118. (canceled)
  • 119. (canceled)
  • 120. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXVIII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is D, E, G, H, K, L, R, or S;X2 is A, F, I, or L;X3 is A, E, F, L, P, Q, R, or S;X4 is E, F, G, M, R, S, V, W, or Y;X5 is E, P, or Q;X6 is A, D, E, or G;X7 is H, L, P, R, S, or T;X8 G, L, S, or Y;X9 is D, H, K, L, T, or Y;X10 is A, F, G, I, L, or R;X11 L, P, or V;X12 is A, E, H, I, K, or R;X13 is E, I, K, N, Q, R, S, or T; andX14 is A, F, H, P, R, S, T, or Y;and(b) a cargo molecule.
  • 121. (canceled)
  • 122. (canceled)
  • 123. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XXXIX): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is A, E, N, Q, R, or W;X2 is A, D, I, L, S, T, or V;X3 is E, H, L, P, T, or Y;X4 is L, S, or W;X5 is A, E, G, V, or Y;X6 is E, G, I, K, P, Q, or T;X7 is H, P, R, or S;X8 is S or Y;X9 is F, S, W, or Y;X10 is A, D, F, G, H, I, T, or V;X11 is L or V;X12 is E, H, K, or N;X13 is H, K, N, or V; andX14 is F, S, T, or Y;and(b) a cargo molecule.
  • 124. (canceled)
  • 125. (canceled)
  • 126. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XL): X1-X2-X3-X4-X5-X6-X7-X8-X9-V-X11-X12-X13-X14, wherein: X1 is D, E, H, L, or Q;X2 is D, F, L, S, or V;X3 is A, H, L, P, Q, or S;X4 is E, G, M, R, or W;X5 is A, D, E, Q, or T;X6 is D or E;X7 is A, L, R, S, or V;X8 is F, L, N, or Y;X9 is F, G, H, L, N, S, or Y;X11 is K, M, or V;X12 is K or T;X13 is D, K, or N; andX14 is F, P, S, or T;and(b) a cargo molecule.
  • 127. (canceled)
  • 128. (canceled)
  • 129. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XL1): X1-X2-X3-X4-X5-X6-X7-X8-X9-V-X11-X12-X13-X14, wherein: X1 is H or Q;X2 is A, F, I, L, T, or V;X3 is H, L, P, R, S, or T;X4 is A, F, G, L, S, W, or Y;X5 is A, D, E, G, N, or T;X6 is A, E, or V;X7 is A, H, P, S, or T;X8 is A, D, H, N, Q, T, or Y;X9 is G, H, L, S, T, V, or Y;X11 is S or V;X12 is K, L, N, P, Q, R, T, or Y;X13 is E, H, I, K, N, or T; andX14 is F or S;and(b) a cargo molecule.
  • 130. (canceled)
  • 131. (canceled)
  • 132. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XLII): X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X1 is H, L, Q, R, or V;X2 is F, G, H, I, L, or V;X3 is A, H, N, P, Q, or T;X4 is F, G, L, M, S, T, or W;X5 is A, D, G, I, K, P, Q, or V;X6 is D, E, G, K, or P;X7 is A, L, P, Q, R, S, or T;X8 is F, H, S, or Y;X9 is A, D, F, G, H, K, N, S, or Y;X10 is I, L, N, or V;X11 is A, E, G, K, L, Q, S, V, or W;X12 is A, D, E, G, I, K, M, N, Q, R, or T;X13 is E, H, K, N, Q, R, T, or V; andX14 is F, H, N, P, S, T, or Y;and(b) a cargo molecule.
  • 133. (canceled)
  • 134. (canceled)
  • 135. A nerve targeting peptide conjugate, comprising (a) a peptide comprising the amino acid sequence of Formula (XLIII): X1-X2-X3-X4-X5-X6-X7-Y-X9-V-X11-X12-X13-X14, wherein X1 is G, H, N, Q, R, or Y;X2 is F, H, I, L, M, or V;X3 is L, P, or Q;X4 is G, L, Q, R, S, or W;X5 is G, H, K, Q, R, or V;X6 is E or V;X7 is P or R;X9 is N, S, Y;X11 is L, Q, or V;X12 is A, E, K, L, or T;X13 is K, M, N, P, Q, or T; andX14 is A, F, I, L, P, S, or Y;and(b) a cargo molecule.
  • 136. (canceled)
  • 137. (canceled)
  • 138. A nerve targeting peptide conjugate, comprising: (a) a peptide comprising the amino acid sequence of Formula (XLIV): Q-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14, wherein: X2 is H, I, M, N, or V;X3 is A, L, T, or V;X4 is F, H, L, N, S, or W;X5 is A, D, K, N, or S;X6 is A, E, H, K, or Q;X7 is P or S;X8 is F, H, N, or Y;X9 is N, S, T, or Y;X10 is G, P, S, V, or Y;X11 is Q or V;X12 is E or K;X13 is D, N, P, R, or T; andX14 is S, T, or Y;and(b) a cargo molecule.
  • 139. (canceled)
  • 140. (canceled)
  • 141. The nerve targeting peptide conjugate of claim 1, wherein the cargo molecule comprises a drug, a fluorescent moiety, or a photosensitizing agent.
  • 142. The nerve targeting peptide conjugate of claim 1, wherein the cargo molecule is joined to the N-terminus or C-terminus of the peptide.
  • 143. The nerve targeting peptide conjugate of claim 1, wherein the cargo molecule is joined to the peptide via a linker.
  • 144.-154. (canceled)
  • 155. A pharmaceutical composition comprising the nerve targeting peptide conjugate of claim 1 and a pharmaceutically acceptable carrier.
  • 156. A method of delivering a cargo molecule to a neuron or nerve comprising contacting the neuron or nerve with the nerve targeting peptide conjugate of claim 1.
  • 157. A method of identifying a neuron or nerve comprising contacting the neuron or nerve with the nerve targeting peptide conjugate of claim 141, wherein the cargo molecule comprises a fluorescent moiety.
  • 158. A method of delivering a photosensitizing agent to a neuron or nerve comprising contacting the neuron or nerve with the nerve targeting peptide conjugate of claim 141, wherein the cargo molecule comprises a photosensitizing agent.
  • 159. (canceled)
  • 160. A method of delivering a drug to a neuron or nerve comprising contacting the neuron or nerve with the nerve targeting peptide conjugate of claim 141, wherein the cargo molecule comprises a drug.
  • 161.-169. (canceled)
CROSS REFERENCE TO RELATED APPLICATION

The present application claims benefit of U.S. Provisional Application No. 63/386,470 filed on Dec. 7, 2022 and is incorporated by reference in its entirety.

Provisional Applications (1)
Number Date Country
63386470 Dec 2022 US