Delivery of molecules across the blood brain barrier

Abstract

A polypeptide linker (2 to 21 amino acids) is connected to a carrier peptide based on the Ioop2 domain of lynx1. The carrier or transporter peptide may be further coupled via the polypeptide linker to a molecule of interest (MOI) or effector agent, such as a protein or protein domain or any other biomolecule (e.g., biomolecules having a molecular weight of 10kDa to 230 kDa). In some embodiments, the complex may be used therapeutically in targeting various neurological disease-causing cellular components.

Description

DELIVERY OF MOLECULES ACROSS THE BLOOD BRAIN BARRIERCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application Serial No. 63 / 646,000, filed May 13, 2024, the entire content of which is incorporated herein by reference.BACKGROUND

[0002] Diminished cognitive abilities are associated with many disease states (e.g., Alzheimer's disease (AD), Parkinson's disease (PD), depression, schizophrenia, and behavioral disorders such as Attention Deficit Hyperactivity Disorder (ADHD)). These disease states have been consistently among the most detrimental to quality of life. While many therapeutic compounds showing potential to alleviate cognitive decline have been identified, to date, effective therapies have proven elusive.

[0003] In the past decades, the cholinergic system, especially the nicotinic acetylcholine receptors, have been shown to be of critical importance for normal cognition. The Ioop2 (also referred to herein as loop II, or the central loop) of the lynxl proteins of the neurotoxin gene family is the putative binding domain of the neuronal nicotinic acetylcholine receptors (nACHRs), as described in U.S. Patent No. 8,629,114, issued January 14, 2014 in the name of Assignee, Ophidion Inc., the entire content of which is incorporated herein by reference. As also described in U.S. Patent No. 8,629,114, the Ioop2 domain may be used as a peptide transporter to deliver payloads to the brain and / or neurons across the blood brain barrier.SUMMARY

[0004] According to embodiments of the present disclosure, a complex comprises a molecule of interest, biomolecule payload, or effector agent; a carrier peptide comprising a peptide represented by SEQ ID NO:1 or SEQ ID NO:34 or variants thereof, and a polypeptide linker linking the carrier peptide to the molecule of interest, the biomolecule payload, or the effector agent.SEQ ID NOUX1 -X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16SEQ ID NO:34X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16In SEQ ID NOs:1 and 34X1 is M or T;X2 is T or I;X3 is T or W;0157444.0804005X4 is R or C;X5 is T or D;X6 is any amino acid, or Y, I, or G;X7 is F or Y;X8 is any amino acid, or T or C;X9 is any amino acid, or P, N, or S;X10 is any amino acid, or Y, T, or S;X12 is M or G;X14 is V or R;X15 is R, S, A, or I; and X16 is K, S, or D.

[0005] In some embodiments, the molecule of interest, biomolecule payload, or effector agent may include a biological molecule. And in some embodiments, the molecule of interest, biomolecule payload, or effector agent may include a biological molecule having a molecular weight of 10 kDa to 230 kDa. For example, in some embodiments, the molecule of interest, biomolecule payload, or effector agent may include an entity selected from antibodies, antibody fragments, proteins, enzymatic protein domains, nucleic acid molecules, virus like particles, full native proteins, protein domains, partial protein domains with a binding affinity or a desired enzymatic activity. In some embodiments, the molecule of interest, biomolecule payload, or effector agent may include an entity selected from nucleic acid molecules such as RNAs and DNAs, aptamers, analogs, combinations of DNA and RNA, genes; peptides including antibody and antigen fragments; proteins, including antibodies, antibodies, engineered antibody like proteins; chemical molecules, viral particles, liposomes; endosomes; exosomes; nanoparticles, dendrimers; positron emission tomograph ligands; eukaryotic cells; prokaryotic cells; microspheres; nanogels; and bionanocapsules.

[0006] In some embodiments, the carrier peptide may include a peptide selected from SEQ ID NOs:1 -7, 9, 10, 13-15, and 34-42. For example, in some embodiments, the carrier peptide may include a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11- X12-K13-X14 (SEQ ID NO:13). In SEQ ID NO:13:X3 is T or W; X4 is R or C; X5 is T or D; X6 is any amino acid; X7 is F or Y;X8 is any amino acid; X9 is any amino acid; X10 is any amino acid; X12 is M or G; and X14 is V or R.0157444.0804005

[0007] In some embodiments, for example, the carrier peptide may include a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11 -X12-K13-X14 (SEQ ID NO: 14). In SEQ ID NO:14:X3 is T or W;X4 is R or C;X5 is T or D;X6 is Y, G or I;X7 is F or Y;X8 is T or C;X9 is P, N, or S;X10 is Y, T, or S;X12 is M or G; and X14 is V.

[0008] According to some embodiments, the carrier peptide may include a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO: 15). In SEQ ID NO:15:X3 is T or W;X4 is R or C;X5 is T or D;X6 is Y, G or I;X7 is F or Y;X8 is T;X9 is P;X10 is Y or T;X12 is M or G; andX14 is V.

[0009] In some embodiments, the polypeptide linker may include a peptide having 2 to 21 amino acids. For example, in some embodiments, the polypeptide linker may include a peptide having 3 to 20 amino acids.

[0010] According to some embodiments, the polypeptide linker may include one or more amino acids selected from S, T, G, A, K, P, and E. In some embodiments, the polypeptide linker may include at least one P residue. And in some embodiments, the polypeptide linker may include (X-Proline)n where X is any amino acid, and n is 1 to 10.

[0011] In some embodiments, the polypeptide linker may include an alpha-helical linker.

[0012] According to some embodiments, the polypeptide linker may include a polypeptide selected from:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)0157444.0804005PQHTGT (SEQ ID NO:47)STDKP (SEQ ID NO:48)SAQSA (SEQ ID NO:49)GGSGT (SEQ ID NQ:50)GSGSG (SEQ ID NO:51 )SGSGN (SEQ ID NO:52)SSTGK (SEQ ID NO:53)GTSDT (SEQ ID NO:54)SSSTG (SEQ ID NO:55)ASGST (SEQ ID NO:56)SKSTS (SEQ ID NO:57)TDGST (SEQ ID NO:58)PGTST (SEQ ID NO:59)SGAQQDSN (SEQ ID NQ:60)APKAPADG (SEQ ID NO:61 )PPASPKADAP (SEQ ID NO:62)SASTKGPS (SEQ ID NO:63)GQPKANPT (SEQ ID NO:64)AGATD (SEQ ID NO:65)NADAT (SEQ ID NO:66)SQSGD (SEQ ID NO:67)SAASSS (SEQ ID NO:68)GKSGK (SEQ ID NO:69)PGTST (SEQ ID NQ:70)PQTDP (SEQ ID NO:71 )PPASP (SEQ ID NO:72)KNTPP (SEQ ID NO:73)GSGSGS (SEQ ID NO:74)GSGSGS (SEQ ID NO:75)GSGSGSGS (SEQ ID NO:76)GSGSGSGSGS (SEQ ID NO:77)GSGSGSGSGSGS (SEQ ID NO:78)GSGSGSGSGSGSGS (SEQ ID NO:79)GSGSGSGSGSGSGSGS (SEQ ID NQ:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81 )GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82) GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)GGSGGSGGSGGS (SEQ ID NO:85)GGSGGSGGSGGSGGS (SEQ ID NO:86)GGSGGSGGSGGSGGSGGS (SEQ ID NO:87)GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88)0157444.0804005GGGS (SEQ ID NO:89)GGGSGGGS (SEQ ID NO:90)GGGSGGGSGGGS (SEQ ID NO:91 )GGGSGGGSGGGSGGGS (SEQ ID NO:92)GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93) EEAEEAEEA (SEQ ID NO:94)EEAEEAEEA (SEQ ID NO:95)EEAEEAEEAEEA (SEQ ID NO:96)EEAEEAEEAEEAEEA (SEQ ID NO:97)EEAEEAEEAEEAEEAEEA (SEQ ID NO:98)EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99)EEAS (SEQ ID NQ:100)EEASEEAS (SEQ ID NQ:101 )EEASEEASEEAS (SEQ ID NQ:102)EEASEEASEEASEEAS (SEQ ID NO: 103)EEASEEASEEASEEASEEAS (SEQ ID NQ:104)XP where X is any amino acidXPXP where X is any amino acidXPXPXP where X is any amino acidXPXPXPXP where X is any amino acid (SEQ ID NO: 105) XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106) XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107) XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108) XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NQ:109) XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110) XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 ) LEGGGGSGGGGSGGGGS (SEQ ID NO:112)GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113) GGGGGGGG (SEQ ID NO:114) GGGGGG (SEQ ID NO:115)EAAAKEAAAKEAAAK (SEQ ID NO:116)EAAAK (SEQ ID NO: 117)EAAAKEAAAK (SEQ ID NO: 118)AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID NO:120)PAPAP (SEQ ID NO:121 )AEAAAKEAAAKA (SEQ ID NO:122)(AP)n where n is 5 to 17APAPAPAPAP (SEQ ID NO: 123)APAPAPAPAPAP (SEQ ID NO: 124)APAPAPAPAPAPAP (SEQ ID NO: 125)0157444.0804005APAPAPAPAPAPAPAP (SEQ ID NO: 126) APAPAPAPAPAPAPAPAP (SEQ ID NO: 127) APAPAPAPAPAPAPAPAPAP (SEQ ID NO: 128) APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 ) APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132) APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR J AETVFPDV (SEQ ID NO: 138)PLG | LWA (SEQ ID NO:139)RVL AEA (SEQ ID NO: 140)EDWCC | SMSY (SEQ ID NO:141 )GGIEGR 1 GS (SEQ ID NO:142)TRHRQPR | GWE (SEQ ID NO:143)AGNRVRR 1 SVG (SEQ ID NO:144)RRRRRRR j R j R (SEQ ID NO:145)GFLGj ((SEQ ID NO:146) valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) mc-ValCitPABC4-(4-acetylphenoxy) butanoic acid Hydrazone mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) mc-PEG-carbonate disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) In the above-listed linkers, “me” is maleimidocaproyl.

[0013] In some embodiments, the polypeptide linker may include a polypeptide selected from:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)PQHTGT (SEQ ID NO:47)STDKP (SEQ ID NO:48)SAQSA (SEQ ID NO:49)GGSGT (SEQ ID NQ:50)GSGSG (SEQ ID NO:51 )SGSGN (SEQ ID NO:52)0157444.0804005SSTGK (SEQ ID NO:53)GTSDT (SEQ ID NO:54)SSSTG (SEQ ID NO:55)ASGST (SEQ ID NO:56)SKSTS (SEQ ID NO:57)TDGST (SEQ ID NO:58)PGTST (SEQ ID NO:59)SGAQQDSN (SEQ ID NQ:60)APKAPADG (SEQ ID NO:61)PPASPKADAP (SEQ ID NO:62)SASTKGPS (SEQ ID NO:63)GQPKANPT (SEQ ID NO:64)AGATD (SEQ ID NO:65)NADAT (SEQ ID NO:66)SQSGD (SEQ ID NO:67)SAASSS (SEQ ID NO:68)GKSGK (SEQ ID NO:69)PGTST (SEQ ID NQ:70)PQTDP (SEQ ID NO:71 )PPASP (SEQ ID NO:72)KNTPP (SEQ ID NO:73)GSGSGS (SEQ ID NO:74)GSGSGS (SEQ ID NO:75)GSGSGSGS (SEQ ID NO:76)GSGSGSGSGS (SEQ ID NO:77)GSGSGSGSGSGS (SEQ ID NO:78)GSGSGSGSGSGSGS (SEQ ID NO:79)GSGSGSGSGSGSGSGS (SEQ ID NQ:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81 )GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82) GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)GGSGGSGGSGGS (SEQ ID NO:85)GGSGGSGGSGGSGGS (SEQ ID NO:86)GGSGGSGGSGGSGGSGGS (SEQ ID NO:87)GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88) GGGS (SEQ ID NO:89)GGGSGGGS (SEQ ID NQ:90)GGGSGGGSGGGS (SEQ ID NO:91 )GGGSGGGSGGGSGGGS (SEQ ID NO:92)GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93) EEA0157444.0804005EEAEEA (SEQ ID NO:94)EEAEEAEEA (SEQ ID NO:95)EEAEEAEEAEEA (SEQ ID NO:96)EEAEEAEEAEEAEEA (SEQ ID NO:97)EEAEEAEEAEEAEEAEEA (SEQ ID NO:98)EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99)EEAS (SEQ ID NQ:100)EEASEEAS (SEQ ID NQ:101 )EEASEEASEEAS (SEQ ID NQ:102)EEASEEASEEASEEAS (SEQ ID NO: 103)EEASEEASEEASEEASEEAS (SEQ ID NQ:104)XP where X is any amino acidXPXP where X is any amino acidXPXPXP where X is any amino acidXPXPXPXP where X is any amino acid (SEQ ID NO: 105)XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106) XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107) XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108) XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:109) XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110) XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 )

[0014] According to some embodiments, the polypeptide linker may include a polypeptide selected from:LEGGGGSGGGGSGGGGS (SEQ ID NO:112)GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113)GGGGGGGG (SEQ ID NO:114)GGGGGG (SEQ ID NO:115)EAAAKEAAAKEAAAK (SEQ ID NO:116)EAAAK (SEQ ID NO: 117)EAAAKEAAAK (SEQ ID NO: 118)AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID NO:120)PAPAP (SEQ ID NO:121 )AEAAAKEAAAKA (SEQ ID NO:122)(AP)n where n is 5 to 17APAPAPAPAP (SEQ ID NO: 1APAPAPAPAPAP (SEQ ID NAPAPAPAPAPAPAP (SEQ IDAPAPAPAPAPAPAPAP (SEQ APAPAPAPAPAPAPAPAP (S APAPAPAPAPAPAPAPAPAP0157444.0804005APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 ) APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132) APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR j AETVFPDV (SEQ ID NO: 138)PLG | LWA (SEQ ID NO:139)RVL AEA (SEQ ID NO: 140)EDWCC | SMSY (SEQ ID NO:141 )GGIEGR 1 GS (SEQ ID NO:142)TRHRQPR | GWE (SEQ ID NO:143)AGNRVRR 1 SVG (SEQ ID NO:144)RRRRRRR j R j R (SEQ ID NO:145)GFLGJ, ((SEQ ID NO:146) valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) mc-ValCitPABC4-(4-acetylphenoxy) butanoic acid Hydrazone mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) m c-P E G -ca rbo n ate disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) In the above-listed linkers, “me” is maleimidocaproyl.

[0015] In some embodiments, the polypeptide linker may include a polypeptide represented by one of the following:(GS)n1(GGS)n2(GGGS (SEQ ID NO:43))n3(EEA)n1(EEAS (SEQ ID NO:44))n2In the above formulae, n1 is 1 -10, n2 is 1 -7, and / or n3 is 1 -5. For example, in some embodiments, n1 may be 1-6, n2 may be 1 -5, and / or n3 may be 1 -4.

[0016] According to some embodiments, the complex may include one carrier peptide and one polypeptide linker coupled to one side of the molecule of interest, biomolecule0157444.0804005 payload, or effector agent, for example at the N terminal side or the C terminal side of a protein or the chemically equivalent side of another non-protein biomolecule.

[0017] In some embodiments, the complex may include a first carrier peptide and a first polypeptide linker coupled to a first side of the molecule of interest, biomolecule payload, or effector agent, for example at the N terminal side of a protein or the chemically equivalent side of another non-protein biomolecule, and a second carrier peptide and a second polypeptide linker coupled to a second side of the molecule of interest, biomolecule payload, or effector agent, for example at the C terminal side of a protein or the chemically equivalent side of another non-protein biomolecule.

[0018] According to some embodiments, the complex may include two carrier peptides, each with a corresponding polypeptide linker, attached to one molecule of interest, biomolecule payload, or effector agent.

[0019] In some embodiments, the molecule of interest, biomolecule payload, or effector agent may be an individual protein or protein fragment, and the N terminus and C terminus may each have a respective carrier peptide and respective polypeptide linker. In some embodiments, each of the respective carrier peptides may have the same binding affinities to neuronal receptors. And in some embodiments, each of the respective carrier peptides may have different binding affinities to neuronal receptors.

[0020] According to some embodiments, the complex may include at least two chimeric complexes, each with at least one respective carrier peptide and respective polypeptide linker, where one or more of the chimeric complexes forms a multimer. In some embodiments, the multimer may be a dimer, trimer, or tetramer. In some embodiments, the multimerization region of the chimeric molecules may be part of the molecule of interest, biomolecule payload, or effector agent, may be at least a portion of the polypeptide linker(s), or may be afforded by added chemical linkers. In some embodiments, the interaction between the chimeric molecules may be covalent.According to some embodiments, the interaction between the chimeric molecules may be noncovalent. In some embodiments, each of the respective carrier peptides may have different binding affinities to neuronal receptors. And in some embodiments, each of the respective carrier peptides may have the same binding affinities to neuronal receptors.

[0021] According to some embodiments, a method of transporting a molecule of interest, biomolecule payload, or effector agent to target cells includes providing a complex disclosed herein to the target cells or to a subject having the target cells.

[0022] In some embodiments, a pharmaceutical composition includes a complex disclosed herein, and one or more pharmaceutically acceptable excipients and / or carriers. According to some embodiments, a method of transporting a molecule of interest, biomolecule payload, or effector agent to target cells includes providing or administering the pharmaceutical composition to the target cells or to a subject having the target cells.0157444.0804005BRIEF DESCRIPTION OF THE DRAWINGS

[0023] FIG. 1 is a schematic representation of a transverse section of the blood brain barrier.

[0024] FIGs. 2A-C depict diagrammatic representations of various formats of peptide- molecule linkages according to embodiments of the present disclosure, with FIG. 2A depicting an N-terminal linkage of a carrier or transporter peptide to a molecule of interest, FIG. 2B depicting a C-terminal linkage of a carrier or transporter peptide to a molecule of interest, and FIG. 2C depicting alternative combinations of dimerization via the molecule of interest and the N- and C- terminal variants.

[0025] FIG. 3 depicts a Western blot analysis of receptor mediated transcytosis.

[0026] FIG. 4 depicts a ball and stick model representation of Ioop2 of theBungarotoxin peptide.

[0027] FIG. 5 is an alignment of full length lynxl amino acid sequences from human (SEQ ID NO:26), mouse (SEQ ID NO:27), macaque (SEQ ID NO:28), cow (SEQ ID NO:29), chimp (SEQ ID NO:30), squirrel monkey (SEQ ID NO:31 ), rat (SEQ ID NO:32), and ferret (SEQ ID NO:33) (top to bottom, respectively) with Ioop2 in bold.DETAILED DESCRIPTION

[0028] As discussed in U.S. Patent No. 8,629,114, the entire contents of which are incorporated herein by reference, lynxl is an accessory molecule to a specific group of neuronal nicotinic acetylcholine receptors, with a striking structural similarity to other nicotinic acetylcholine receptor binding-proteins, such as a-bungarotoxin, SLURPs, (Secreted Ly-6 / uPAR-related proteins), and prostate stem cell antigen. The lynxl protein has been shown to bind to neuronal nicotinic receptors (NNRs) or nicotinic acetylcholine receptors (nAChRs), as disclosed in Ibanez-Tallon et al., 2002 Neuron, 33:893-903, the entire contents of which are incorporated herein by reference. As described in U.S. Patent No. 8,629,114, the Ioop2 region of lynxl can be used as a peptide transporter for delivery of a therapeutic agent across the blood brain barrier (BBB).

[0029] According to embodiments of the present disclosure, compositions and methods for delivery across the BBB include polypeptide linkers connected to a carrier peptide (also referred to herein, interchangeably, as a transporter peptide or peptide transporter) capable of crossing the BBB, and to a shuttled protein or biomolecule (also referred to herein as a biomolecule payload or a molecule of interest (MOI)). These compositions and methods can deliver biological molecules across the BBB through specific subtypes of the nicotinic acetylcholine receptor. Nonlimiting examples of the biological molecules (or payloads) that can be delivered according to embodiments of the0157444.0804005 present disclosure include antibodies, antibody fragments, proteins, enzymatic protein domains, nucleic acid molecules, virus like particles, etc.

[0030] According to embodiments of the present disclosure, the polypeptide linkers can enhance the effectiveness of the carrier or transporter peptide when coupled to the MOI or biomolecule payload, and / or can enhance stabilization of the MOI. And in some embodiments, the polypeptide linkers may be effective in enhancing transportation of the complex (i.e. , the carrier or transporter peptide linked to the MOI via the polypeptide linker) across the blood brain barrier (BBB) via a nicotinic acetylcholine receptor mediated transcytosis.

[0031] Abbreviations for amino acids are used throughout this disclosure and follow the standard nomenclature known in the art. For example, as would be understood by those or ordinary skill in the art, Alanine is Ala or A; Arginine is Arg or R; Asparagine is Asn or N; Aspartic Acid is Asp or D; Cysteine is Cys or C; Glutamic acid is Glu or E; Glutamine is Gin or Q; Glycine is Gly or G; Histidine is His or H; Isoleucine is He or I; Leucine is Leu or L; Lysine is Lys or K; Methionine is Met or M; Phenylalanine is Phe or F; Proline is Pro or P; Serine is Ser or S; Theonine is Thr or T; Tryptophan is Trp or W; Tyrosine is Tyr or Y; and Valine is Vai or V.

[0032] In some embodiments of the present invention, the carrier peptides when linked to a biomolecule payload or MOI are capable of crossing the blood brain barrier and delivering the biomolecule payload or MOI into the brain and / or neurons.

[0033] As used herein, a “fragment” of a polynucleotide or polypeptide refers to a smaller set of nucleotides or peptides with respect to the referenced polynucleotide or polypeptide.

[0034] As used herein, a “variant” of a polynucleotide or polypeptide, refers to a polynucleotide or polypeptide that can vary in primary, secondary, or tertiary structure, as compared to the referenced polynucleotide or polypeptide. For example, a peptide fragment of a Iynx1-loop2 peptide is a peptide that may refer to the entire peptide sequence of Iynx1 -loop2, or a shorter peptide sequence, and this peptide fragment may include amino acid substitutions or deletions as described herein. For example, a “variant” of a Iynx1 -loop2 peptide refers to a molecule substantially similar in structure and function to the referenced Iynx1-loop2 peptide, but may include additional moieties or substitutions or changes. As such, a variant peptide may include a derivative which is the referenced peptide that has been chemically modified, for example by techniques such as ubiquitination, labeling, pegylation (derivatization with polyethylene glycol) or addition of other molecules. A molecule is said to be “substantially similar” to another molecule if both molecules have substantially similar structures or if both molecules possess a similar biological activity. Thus, provided that two molecules possess a similar activity, they are considered variants as that term is used herein even if the structure of one of the0157444.0804005 molecules is not found in the other, or if the sequence of amino acid residues is not identical.

[0035] As used herein, fragments, variants and substantially similar molecules of Iynx1 -loop2, are collectively referred to as “Iynx1 -loop2-derived.”

[0036] As used herein, “biomolecule payload” and “molecule of interest” (or “MOI”) are used interchangeably, and refer to any biological molecule that imparts an effect on a target inside the blood brain barrier. As disclosed herein, the target may be target cells or extracellular molecules. Non-limiting examples of biomolecule payloads or MOIs that can be linked to a carrier peptide according to embodiments of the present disclosure include antibodies, antibody fragments, proteins, enzymatic protein domains, nucleic acid molecules, virus like particles, etc. For example, in some embodiments, non-limiting examples of some MOIs include previously characterized molecules, such as agonistic and antagonistic proteins including but not limited to antibodies and antibody fragments. Also, the MOI can, but need not include full native proteins, and protein domains can also be used (for example, a partial domain with a binding affinity or a particular desired enzymatic activity). The molecular weight of the biological molecule of the MOI or biomolecule payload is not particularly limited, but in some embodiments, may be 10kDa to 230 kDa.

[0037] Moreover, the term “MOI” refers not only to the biomolecule payloads discussed above, but also to any effector agent that imparts an effect on a target inside the BBB. Non-limiting examples of such effector agents include the biomolecule payloads discussed herein, as well as the effector agents discussed in U.S. Patent No. 8,629,114. Some specific non-limiting examples of MOIs may include nucleic acid molecules such as RNAs and DNAs, aptamers, analogs, combinations of DNA and RNA, genes; peptides including antibody, antigen fragments; proteins, including antibodies, antibodies, engineered antibody like proteins; chemical molecules, viral particles, liposomes; endosomes; exosomes; nanoparticles, dendrimers; positron emission tomograph ligands; eukaryotic cells; prokaryotic cells; microspheres; nanogels; and bionanocapsules. Some specific and non-limiting examples of such effector agents include siRNA; short hairpin or stem loop RNA (shRNA); microRNA, double stranded RNA (dsRNA); strand template RNA (stRNA); oligonucleotides (DNA or RNA); modified oligonucleotides (DNA or RNA); aptamers; analogs and combinations of DNA and RNA; genes; peptides including antibody and antigen fragments; proteins, including antibodies and antigens; small chemical molecules; large chemical molecules; viral particles; liposomes; endosomes; exosomes; nanoparticles; dendrimers (e.g. Poly(amidoamine), or PAMAM); positron emission tomography (PET) ligands; eukaryotic cells; prokaryotic cells; microspheres; nanogels; and bionanocapsules. The target may also be any suitable target, non-limiting examples of which include those “targets” discussed in U.S. Patent No. 8,629,114, and may be, for example, target cells or extracellular molecules.0157444.0804005

[0038] As used herein, “linked” refers to the attachment of two or more entities via a linker to form one entity or “conjugate.” For example, embodiments of the present disclosure reference linkage of a carrier peptide to a biomolecule payload via a linker. The linkage can be permanent or reversible. In some embodiments, several linkers can be included in order to take advantage of desired properties of each linker and each component in the conjugate. Peptide linkers can be linked by expressing DNA encoding the linker to one or more proteins in the conjugate. Linkers can be enzymatically cleavable, acid cleavable, photocleavable, and heat sensitive linkers. Any method of effecting the linkage or conjugation may be used.

[0039] As used herein, “target” refers to a cell or extracellular molecule which is entirely within the BBB-protected central nervous system (CNS) tissue. Extracellular molecules include, but are not limited to extracellular proteins and tissues. An example of extracellular proteins within the BBB, include [3-amyloid plaques characteristic of Alzheimer's disease (AD), as disclosed in Cramer et al., Science, 335:1503, 2012, the entire contents of which are incorporated herein by reference. In addition to AD, Parkinson's disease and Lewy body dementias are characterized by misfolded protein aggregates called Lewy bodies, and one of the major components of these bodies is the a-synuclein protein, as disclosed in Clinton et al., J. Neurosci, 30:7281-7289, 2010, the entire contents of which are incorporated herein by reference.

[0040] The term “target cells” as used herein also refers to cells expressing the alpha (a) subunit and / or the beta (P) subunit of the nicotinic acetylcholine receptor. A carrier peptide as disclosed herein binds to the alpha-subunit of the nicotinic acetylcholine receptor. Accordingly, a carrier peptide as disclosed herein is useful as a targeting moiety for the selective targeting of cells expressing the alpha subunit of the nicotinic acetylcholine receptor. Cells expressing the alpha (a) subunit of the nicotinic acetylcholine receptor include, for example, neurons, glial cells and endothelial cells comprising the blood brain barrier. Target cells also include cells whose endogenous milieu is separated by the BBB, for example, cells in the central nervous system, e.g., brain cells, spinal cord cells, glial cells, and other cells supporting neurons, for e.g. astrocytes or “nursing cells” and cells of the central nervous system. In some embodiments, the target cells can be any cell expressing the alpha subunit of the nicotinic acetylcholine receptor or a homologue thereof, such as, for example, but not limited to, neuronal cells in a subject (i.e. in vivo), neuronal cells ex vivo or cultured neuronal cells (i.e. in vitro) such as, for example, as primary neuronal cultured cells, or immortalized cell lines expressing alpha and / or beta subunits of the nicotinic acetylcholine receptor either naturally or through stable selection of transfected alpha and / or beta nicotinic acetylcholine receptor constructs. In some embodiments, the target cells are neuronal precursor or neuronal progenitor cells, such as neuronal progenitor stem cells that express an alpha subunit of the nicotinic acetylcholine receptor or a homologue thereof. In some embodiments of the present invention, the target is present within a subject, for example a mammalian subject, for example a human subject. In alternative embodiments, the target is ex vivo, and in further embodiments, the target is in a biological sample, for example, in vitro.0157444.0804005

[0041] According to some embodiments of the present disclosure, a composition or complex capable of crossing the blood brain barrier includes at least one carrier peptide, at least one linker, and at least one biomolecule payload (or MOI). In some embodiments, the carrier peptide is coupled to the linker, thereby enhancing the desired cellular uptake and / or transport of the MOI across the BBB.

[0042] The linkers according to embodiments of the present disclosure have particular sequence(s) and structural configurations that enhance the ability of the connected carrier peptide to effectively bind to the nicotinic acetylcholine receptor when the composition also includes a biomolecule payload (or MOI). The linker(s) may be connected to the biomolecule payload in any suitable manner. For example, in some embodiments, the linker(s) may be connected to either the N-term and / or C-term of proteins, or the 3’ or 5’ end of nucleic acids. According to some embodiments, therefore, a unique binding moiety can be engineered or tailored based on the desired biomolecule payload, and the composition (i.e. , the linker, carrier peptide, and biomolecule payload complex) will bind to a region in the nicotinic acetylcholine receptor. As a result of such binding, the complex undergoes receptor-mediated transcytosis to deliver the biomolecule payload across the BBB.

[0043] In some embodiments, the biomolecule payload may be connected to a carrier peptide via the linker at only one terminus of the MOI. However, in some embodiments, the MOI may be connected to carrier peptide(s) via the linker(s) at each terminus of the MOI so that the resulting complex displays two separate binding motifs. Such arrangements may allow for multiple specificities to subtypes of the nicotinic acetylcholine receptors.

[0044] The carrier peptide (or transporter peptide) may be any of the transporter peptides described in U.S. Patent No. 8,629,114, the entire content of which is incorporated herein by reference. Indeed, the carrier peptide (or transporter peptide) may include any of the Iynx1 -loop2-derived peptides described in U.S. Patent No. 8,629,114, and the carrier peptide may be derived from an individual native origin or a combination of Iynx1 -loop2 domains. For example, in some embodiments, the Iynx1-loop2 derived carrier peptide may be a 12-mer or longer, 16-mer or longer, or 17-mer or longer peptide having any of the general carrier peptide sequence(s) described in U.S. Patent No. 8,629,114.

[0045] The carrier peptide according to embodiments of the present disclosure may also include the possible mutants or include individual substitutions, deletions or additions of about 1 to 5 amino acids, as described in U.S. Patent No. 8,629,114. The substitutions may depend on the conservative nature of the amino acid which may be based upon an existing amino acid’s exposure to the solvent which may define the selection of the substitutions, which is well known in the art, and which is also discussed in U.S. Patent No. 8,629,114.0157444.0804005

[0046] In some embodiments, the carrier peptide may also be modified to accommodate specific MOIs. This may include modifying the amino terminus, for example, by addition of polar group(s) to increase hydrophilicity, as also described in U.S. Patent No. 8,629,114.

[0047] More specifically, as described in U.S. Patent No. 8,629,114, and according to embodiments of the present disclosure, the carrier peptide may be a 16-mer peptide having the general sequence of X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16, (SEQ ID NO: 1 ) in which for SEQ ID NO:1 :Xi is M or T;X2 is T or I;X3is T or W;X4 is R or C;Xs is T or D;X6is Y, I, or G;X7is F or Y;Xs is T or C;Xg is P, N, or S;X10 is Y, T, or S;X12 is M or G;X14 is V or R;X15 is R, S, A, or I; and X16 is K, S, or D.

[0048] Nonlimiting examples of carrier peptides satisfying SEQ ID NO:1 include MTTRTYFTPYRMKVRK (SEQ ID NO:2), MTTRTYYTPTRMKVSK (SEQ ID NO:3), and MTTRTYFTPYRGKVRK (SEQ ID NO:5).

[0049] Alignment of the human lynxl -Ioop2 domain with the lynxl -Ioop2 sequences from mouse, macaque, cow, chimp, squirrel monkey, rat, and ferret is shown in FIG. 5. In some embodiments of the present disclosure, the carrier peptide may have a sequence that represents that which is conserved or similar to the aligned sequences found in various species as shown in FIG. 5. Non-limiting examples of such carrier peptides that are capable of crossing the BBB include: MTTRTYFTPYRMKVRK (SEQ ID NO:2), MTTRTYYTPTRMKVSK (SEQ ID NO:3), MTWCDYFTPSRGKVRKS (SEQ ID NO:4), or MTTRTYFTPYRGKVRK (SEQ ID NO:5).

[0050] According to some embodiments, the carrier peptide sequence(s) may or may not fully satisfy the formula of SEQ ID NO:1 , and may have more or fewer residues than the 16-mer of SEQ ID NO: 1 . For example, as noted above, in some embodiments, the carrier peptide may be a 16-mer (as shown and described in SEQ ID NO:1 ), and in some embodiments, the carrier peptide may be a 17-mer (such as, for example, SEQ ID NO:4). In some embodiments, for example, the carrier peptide may be an 11 -mer to 21 -mer peptide, including 11-mer, 12-mer, 13-mer, 14-mer, 15-mer, 16-mer, 17-mer, 18-mer, 19-0157444.0804005 mer, 20-mer, or 21 -mer peptide having a sequence according to SEQ ID NO:1 in which 1 to 5 residues have been either added or deleted from the SEQ ID NO:1 sequence. In some embodiments, for example, the 1 to 5 residues may be added or deleted from either terminal end of the SEQ ID NO:1 sequence, or one or more may be added or deleted to or from one terminal end, and another one or more may be added or deleted to or from the other terminal end. A non-limiting example of one such mutant includes SEQ ID NO:4 (MTWCDYFTPSRGKVRKS), which includes an added terminal S residue.

[0051] However, in some embodiments, mutants of the SEQ ID NO:1 sequence may additionally or alternatively include substitutions, additions, or deletions in other areas of the sequence. The choice of conservative amino acids for conservative substitutions, or the choice of which amino acids to add or delete within the sequence, may be selected based on the location of the amino acid to be substituted, added, or deleted in the peptide, for example if the amino acid is on the exterior of the peptide and exposed to solvents, or on the interior and not exposed to solvents.

[0052] With respect to substitutions, for example, according to some embodiments, one can select an amino acid which will substitute an existing amino acid based on the location of the existing amino acid, i.e. its exposure to solvents (i.e. if the amino acid is exposed to solvents or is present on the outer surface of the peptide or polypeptide as compared to internally localized amino acids not exposed to solvents). Selection of conservative amino acid substitutions are well known in the art, for example as disclosed in Dordo et al, J. Mol. Biol, 1999, 217, 721-739 and Taylor et al, J. Theor. Biol. 119 (1986);205-218 and S. French and B. Robson, J. Mol. Evol. 19 (1983)171. Accordingly, one can select conservative amino acid substitutions suitable for amino acids on the exterior of a protein or peptide (i.e. amino acids exposed to a solvent), for example, but not limited to, the following substitutions can be used: substitution of Y with F; T with S, K, or A; P with A; E with D or Q; N with D or G; R with K; G with N or A; T with S, K, or A; D with N or E, I with L or V, F with Y or L; S with T or A, R with K, G with N or A, K with R; A with S, K, P, G, T, or V; Wwith Y; and M with L.

[0053] Some non-limiting examples of suitable such mutants include: X1X2X3MPENPRPGTPX4X5X6X7X8X9X10RX12KX14X15X16 (SEQ ID NO:9) in which the sequence of MPENPRPGTP (SEQ ID NO:8) is added between residues X3 and X4of SEQ ID NO:1 ;MTTMPENPRPGTPRTYFTPYRMKVRKS (SEQ ID NO: 10) in which MPENPRPGTP (SEQ ID NO: 8) is inserted between residues X3 and X4 of an example SEQ ID NO:1 sequence;MTTRTYATPYRMKVRKS (SEQ ID NO:6) in which F at position 7 is substituted with A; andMTTRTYFTPYAMADRKS (SEQ ID NO:7) in which the RMKV (SEQ ID NO:22) residues at positions 11 -14 are substituted with AMAD (SEQ ID NO:23).0157444.0804005

[0054] According to some embodiments, taking into account some of these mutant considerations (i.e., substitution, addition, and / or deletion considerations), the carrier peptide may be represented by SEQ ID NO: 1 in which residues Xs, Xs, X9 and X10 may instead be any amino acid, and / or in which the SEQ ID NO: 1 sequence may include up to an additional 5 amino acids, each of which may be individually appended to either end of the peptide. And in some embodiments, the carrier peptide may be represented by SEQ ID NO: 1 in which residues Xe, Xs, X9 and X10 may instead be any amino acid, and / or in which the SEQ ID NO:1 sequence may omit or delete up to 5 amino acids, each of which may be individually omitted from either end of the peptide. In some embodiments, for example, such a smaller peptide may be represented by X3-X4-X5-X6-X7-X8-X9-X10-R11- Xi2-Ki3-Xi4 (SEQ ID NO: 13) in which for SEQ ID NO: 13:X3is T or W;X4 is R or C;Xs is T or D;Xs is any amino acid;X7is F or Y;Xs is any amino acid;X9 is any amino acid;X10 is any amino acid;X12 is M or G; and X14 is V or R.

[0055] For example, in some embodiments, non-limiting examples of such sequences include those represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K-X14 (SEQ ID NO:14) in which for SEQ ID NO:14:X3is T or W;X4 is R or C; Xs is T or D; X6is Y, G or I; X7is F or Y; Xs is T or C; Xg is P, N, or S; X10 is Y, T, or S; X12 is M or G; and X14 is V.

[0056] Additional non-limiting examples of such sequences include those represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:15) in which for SEQ ID NO: 15: X3is T or W; X4 is R or C;Xs is T or D;X6is Y, G or I;X7is F or Y;Xs is T;0157444.0804005X9is P;X10 is Y or T;X12 is M or G; and X14 is V.

[0057] According to some embodiments, the carrier peptide may be any of the carrier peptides or a variant thereof disclosed herein and in U.S. Patent No. 8,629,114. For example, in some embodiments, the carrier peptide may be a variant or mutant of SEQ ID NO:1 in which the first residue (Xi) is deleted. These variant carrier peptides may be represented by X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16, (SEQ ID NO:34) in which for SEQ ID NO:34:X2 is T or I;X3is T or W;X4 is R or C; Xs is T or D; X6is Y, I, or G; X7is F or Y; Xs is T or C; X9 is P, N, or S; X10 is Y, T, or S; X12 is M or G; X14 is V or R; X15 is R, S, A, or I; and X is K, S, or D.

[0058] According to some embodiments, taking into account some of mutant or variant considerations discussed herein (i.e., substitution, addition, and / or deletion considerations), the variant carrier peptide may be represented by SEQ ID NO:34 in which residues Xe, Xs, X9 and X10 may instead be any amino acid, and / or in which the SEQ ID NO:34 sequence may include up to an additional 5 amino acids, each of which may be individually appended to either end of the peptide. And in some embodiments, the carrier peptide may be represented by SEQ ID NO:34 in which residues Xe, Xs, X9and X10 may instead be any amino acid, and / or in which the SEQ ID NO:34 sequence may omit or delete up to 5 amino acids, each of which may be individually omitted from either end of the peptide. Accordingly, the SEQ ID NO:34 peptides and variants or mutants thereof may include a 10-mer to 20-mer peptide. Also, it is understood that the peptides represented by SEQ ID NOs:13-15 - all of which omit the Xi residue from the base SEQ ID NO:1 sequence - are non-limiting examples of both variants or mutants of SEQ ID NO:1 as well as variants or mutants of SEQ ID NO:34.

[0059] The functional structure of such carrier peptides according to SEQ ID NO:34 with the first residue from SEQ ID NO:1 deleted include a beta turn between the first residue and the last residue, as shown in FIG. 4. For example, in some embodiments, an Arg (R) at position 11 (in the formula of SEQ ID NO:1 , or position 10 in SEQ ID NO:340157444.0804005 when considering the deleted first residue) may be buried deep into the acetylcholine receptor binding site. And a Phe (F), for example, at position 7 of SEQ ID NO:1 (or position 6 in SEQ ID NO:34 when considering the deleted first residue) may form a cation- pi interaction with the Arg (R) at position 11 of SEQ ID NO:1 (or position 10 of SEQ ID NO:34 when considering the deleted first residue) to further stabilize the interaction.

[0060] Non-limiting examples of suitable such variant carrier peptides include: TTRTYFTPYRMKVRK (SEQ ID NO:35);TTRTYYTPTRMKVSK (SEQ ID NO:36);TWCDYFTPSRGKVRKS (SEQ ID NO:37);TTRTYFTPYRGKVRK (SEQ ID NO:38);X2X3MPENPRPGTPX4X5X6X7X8X9X10RX12KX14X15X16 (SEQ ID NO:39) in which the sequence of MPENPRPGTP (SEQ ID NO: 8) is added between residues X3 and X4 of SEQ ID NO:34;TTMPENPRPGTPRTYFTPYRMKVRKS (SEQ ID NQ:40) in which MPENPRPGTP (SEQ ID NO: 8) is inserted between residues X3 and X4 of an example SEQ ID NO:34 sequence;TTRTYATPYRMKVRKS (SEQ ID NO:41 ) in which F at position 6 in SEQ ID NO:34 is substituted with A; andTTRTYFTPYAMADRKS (SEQ ID NO:42) in which the RMKV (SEQ ID NO:22) residues at positions 10-13 are substituted with AMAD (SEQ ID NO:23).

[0061] As discussed herein, the compositions or complexes according to embodiments of the present disclosure also include a linker in combination with the carrier peptide. In some embodiments, the linker may be a polypeptide linker having 2 to 21 amino acids, for example 2 to 20, or 3 to 20 amino acids. In some embodiments, the linker may be longer, for example up to about 50 amino acids.

[0062] Table 1 below lists some formulae for non-limiting examples of suitable carrier peptide-linker complexes according to embodiments of the present disclosure.0157444.0804005

[0063] As noted in Table 1 , in some embodiments, exemplary N-terminal and C- terminal linkers may be flexible, and non-limiting examples of which include GS linkers, such as those described in Argos, J Mol Biol. 211 :943 (1990), the entire content of which is incorporated herein by reference. Other flexible linkers may also be used, and may be composed of other small polar amino acids such as Serine or Threonine, and the solubility may be increased by utilizing Threonine and Alanine to maintain flexibility, and Lysine and Glutamic Acid to increase solubility.

[0064] And in some embodiments, exemplary N-terminal linkers may be rigid, and nonlimiting examples of which include alpha-helical linkers, such as those described in George and Heringa, Protein Eng. 15:871 (2002), the entire content of which is incorporated herein by reference. Other rigid linkers may also be used, and may employ Proline rich sequences such as (X-Proline)n where X is any amino acid, and n is 1 to 10.

[0065] Additionally, in some embodiments, in vivo cleavable linkers may be used, for example, when the functional domains are desired to be separated in vivo. Common in vivo cleavable linkers utilize disulfide bonding, such as disclosed in Chen et al., Biotechniques. 49:513 (2010); and Zhao et al., Mol Pharm. 9:664 (2012)), the entire contents of all of which are incorporated herein by reference. Such additions may be used to form multimeric chimeric molecules (or complexes) or to form any other attachments to MOIs (or biomolecule payloads). Additional suitable cleavable linkers include furin and thrombin cleavable linkers, nonlimiting examples of which include RKKRR (SEQ ID NO: 137), RXK (where X is any amino acid), and RR.

[0066] Additional non-limiting examples of suitable linkers include those described in Chen et al., “Fusion Protein Linkers: Property, Design and Functionality,” Adv. Drug Deliv Rev., 65(10): 1357-1369 (2013); and Balamkundu et al., “Lysosomal-Cleavable Linkers in Antibody-Drug Conjugates,” Biomedicines, 11 (11 ), 3080 (2023), the entire contents of all of which are incorporated herein by reference.

[0067] Non-limiting examples of suitable linkers for either the N-terminus or C-terminus include:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)PQHTGT (SEQ ID NO:47)STDKP (SEQ ID NO:48)SAQSA (SEQ ID NO:49)GGSGT (SEQ ID NQ:50)GSGSG (SEQ ID NO:51 )SGSGN (SEQ ID NO:52)SSTGK (SEQ ID NO:53)GTSDT (SEQ ID NO:54)SSSTG (SEQ ID NO:55)ASGST (SEQ ID NO:56)0157444.0804005SKSTS (SEQ ID NO:57)TDGST (SEQ ID NO:58)PGTST (SEQ ID NO:59)SGAQQDSN (SEQ ID NQ:60)APKAPADG (SEQ ID NO:61 )PPASPKADAP (SEQ ID NO:62)SASTKGPS (SEQ ID NO:63)GQPKANPT (SEQ ID NO:64)AGATD (SEQ ID NO:65)NADAT (SEQ ID NO:66)SQSGD (SEQ ID NO:67)SAASSS (SEQ ID NO:68)GKSGK (SEQ ID NO:69)PGTST (SEQ ID NQ:70)PQTDP (SEQ ID NO:71 )PPASP (SEQ ID NO:72)KNTPP (SEQ ID NO:73)GSGSGS (SEQ ID NO:74)GSGSGS (SEQ ID NO:75)GSGSGSGS (SEQ ID NO:76)GSGSGSGSGS (SEQ ID NO:77)GSGSGSGSGSGS (SEQ ID NO:78)GSGSGSGSGSGSGS (SEQ ID NO:79)GSGSGSGSGSGSGSGS (SEQ ID NQ:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81)GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82) GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)GGSGGSGGSGGS (SEQ ID NO:85)GGSGGSGGSGGSGGS (SEQ ID NO:86)GGSGGSGGSGGSGGSGGS (SEQ ID NO:87)GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88) GGGS (SEQ ID NO:89)GGGSGGGS (SEQ ID NQ:90)GGGSGGGSGGGS (SEQ ID NO:91 )GGGSGGGSGGGSGGGS (SEQ ID NO:92)GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93) EEAEEAEEA (SEQ ID NO:94)EEAEEAEEA (SEQ ID NO:95)EEAEEAEEAEEA (SEQ ID NO:96)EEAEEAEEAEEAEEA (SEQ ID NO:97)0157444.0804005EEAEEAEEAEEAEEAEEA (SEQ ID NO:98)EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99)EEAS (SEQ ID NQ:100)EEASEEAS (SEQ ID NQ:101 )EEASEEASEEAS (SEQ ID NQ:102)EEASEEASEEASEEAS (SEQ ID NO: 103)EEASEEASEEASEEASEEAS (SEQ ID NQ:104)XP where X is any amino acidXPXP where X is any amino acidXPXPXP where X is any amino acidXPXPXPXP where X is any amino acid (SEQ ID NO: 105)XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106) XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107) XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108) XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:109) XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110) XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 )

[0068] Additional non-limiting examples of suitable linkers for either the N-terminus or C-terminus include:LEGGGGSGGGGSGGGGS (SEQ ID NO:112)GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113)GGGGGGGG (SEQ ID NO:114)GGGGGG (SEQ ID NO:115)EAAAKEAAAKEAAAK (SEQ ID NO:116)EAAAK (SEQ ID NO: 117)EAAAKEAAAK (SEQ ID NO: 118)AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID NO:120)PAPAP (SEQ ID NO:121 )AEAAAKEAAAKA (SEQ ID NO:122)(AP)n where n is 5 to 17APAPAPAPAP (SEQ ID NO: 123)APAPAPAPAPAP (SEQ ID NO: 124)APAPAPAPAPAPAP (SEQ ID NO: 125)APAPAPAPAPAPAPAP (SEQ ID NO: 126)APAPAPAPAPAPAPAPAP (SEQ ID NO: 127) APAPAPAPAPAPAPAPAPAP (SEQ ID NO: 128) APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 ) APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132)0157444.0804005APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)

[0069] Additional non-limiting examples of suitable cleavable linkers for either the N- terminus or C-terminus include:RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR J AETVFPDV (SEQ ID NO: 138) - Factor Xla / FVI la sensitive cleavagePLG J, LWA (SEQ ID NO:139) - Matrix metalloprotease-1 sensitive cleavage RVL J, AEA (SEQ ID NO: 140) - Matrix metalloprotease-1 sensitive cleavage EDWCC J, SMSY (SEQ ID NO:141 ) - Matrix metalloprotease-1 sensitive cleavageGGIEGR J. GS (SEQ ID NO: 142) - Matrix metalloprotease-1 sensitive cleavage TRHRQPR | GWE (SEQ ID NO:143) - HIV PR (HIV-1 protease cleavage AGNRVRR | SVG (SEQ ID NO: 144) - NS3 protease (HCV protease) cleavage RRRRRRR j R j R (SEQ ID NO: 145) - Factor Xa sensitive cleavage GFLGJ, ((SEQ ID NO: 146) - Furin sensitive cleavage valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) - lysosomal cleavage mc-ValCitPABC - lysosomal cleavage 4-(4-acetylphenoxy) butanoic acid - pH sensitive cleavage Hydrazone - pH sensitive cleavage mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) - lysosomal degradation cleavage mc-P EG -carbon ate - pH cleavage disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) - glutathione cleavable* “me” refers to maleimidocaproyl

[0070] All peptides disclosed herein, including the carrier peptides and linker peptides may be constructed by any appropriate method known in the art. For example, peptides may be synthesized using a peptide synthesizer (Applied Biosystems Model 433) or can be synthesized recombinantly by methods well known in the art. Methods and materials for chemical synthesis of polypeptides are well known in the art. See, e.g., Merrifield, 1963, "Solid Phase Synthesis," J. Am. Chem. Soc. 83:2149-2154. The method by which a peptide of the present invention is synthesized does not limit the present invention. For example, peptides may be commercially purchased from, for example, LifeTein, LLC, or Bio-Synthesis, Inc.0157444.0804005

[0071] The peptides according to embodiments of the present disclosure, including the carrier peptides and the linker peptides, may be modified to accommodate particular target effects. For example, peptides may be modified to accommodate specific biomolecule payloads or MOIs. For example, a carrier peptide can be modified at the amino terminus, for example, so as to increase hydrophilicity. Increased hydrophilicity enhances exposure of the peptides on the surfaces of lipid-based carriers into which the parent peptide-lipid conjugates have been incorporated for use with liposomes or lipid carriers. Polar groups suitable for attachment to peptides so as to increase their hydrophilicity are well known, and include, for example and without limitation: acetyl ("Ac"), 3-cyclohexylalanyl ("Cha"), acetyl-serine ("Ac Ser"), acetyl-seryl-serine ("Ac-Ser- Ser-"), succinyl ("Sue"), succinyl-serine ("Suc-Ser"), succinyl-seryl-serine ("Suc-Ser-Ser"), methoxy succinyl ("MeO-Suc"), methoxy succinyl-serine ("MeO-Suc-Ser"), methoxy succinyl-seryl-serine ("MeO-Suc-Ser-Ser") and seryl-serine ("Ser-Ser-") groups, polyethylene glycol ("PEG"), polyacrylamide, polyacrylomorpholine, polyvinylpyrrolidine, a polyhydroxyl group and carboxy sugars, e.g., lactobionic, N-acetyl neuraminic and sialic acids, groups. The carboxy groups of these sugars would be linked to the N-terminus of the peptide via an amide linkage. One non-limiting example of an N-terminal modification is a methoxy-succinyl modification.

[0072] The complexes or compositions (also referred to herein as a chimeric molecule) according to embodiments of the present disclosure, may have any of various configurations, some non-limiting examples of which are shown in FIGs. 2A-2C. In some embodiments, for example, the chimeric molecule (or complex) may have one carrier peptide and linker coupled to the MOI at the N term side of a protein (or the chemically equivalent side of another non-protein biomolecule) (See FIG. 2A). In some embodiments, one carrier peptide and linker may be coupled to the C-term side of a protein (or the chemically equivalent side of another non-protein biomolecule) (See FIG. 2B)

[0073] In some embodiments, as shown in FIG. 2C, the chimeric molecule (or complex) may have two carrier peptides, each with a linker, and attached to one MOI. In embodiments in which the MOI is an individual protein or protein fragment, the N term and C term may each have a carrier peptide and a linker, and the carrier peptides may have the same binding affinities to neuronal receptors or different binding affinities.

[0074] In some embodiments, there may be at least two chimeric molecules (or complexes), each with at least one carrier peptide and linker. One or more of the chimeric molecules (or complexes) may form multimers, non-limiting examples of which include dimers (such as those shown in FIG. 2C), trimers, and tetramers. The multimerization region of the chimeric molecules may be part of the MOIs, at least a portion of the polypeptide linker(s), or afforded by added chemical linkers. The interaction between the chimeric molecules may be covalent or noncovalent. The carrier peptides may have the same binding affinities to neuronal receptors or different binding affinities.0157444.0804005

[0075] The complexes or compositions according to embodiments of the present disclosure are capable of crossing the blood brain barrier and targeting the MOI (or effector agent) for delivery to the target cell and / or molecule. The MOI (or effector agent) is described above. As described above, in some embodiments, the MOI may include a biological molecule. In such embodiment, the molecular weight of the biological molecule of the MOI or biomolecule payload is not particularly limited, but in some embodiments, may be 10kDa to 230 kDa.

[0076] In some embodiments of the present disclosure, the MOI or effector agent, for example an siRNA therapeutic agent as disclosed herein, can be prepared to be delivered in a “prodrug” form. As would be understood by those skilled in the art, the term “prodrug” indicates a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e. , drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and / or conditions.

[0077] In some embodiments, the MOI or effector agent may be transported to various target cells or tissues. For example, the MOI or effector agent may be transported to any nerve cell, e.g. a nerve cell in the central nervous system, olfactory, or visual system. The MOI or effector agent may also be transported to a neurologically related target cell or tissue, e.g. cells or tissues that interact with or are targets of the nervous system.

[0078] As used herein, the term “gene” refers to a nucleic acid comprising an open reading frame encoding a polypeptide, including both exon and (optionally) intron sequences. A “gene” refers to coding sequence of a gene product, as well as non-coding regions of the gene product, including 5'UTR and 3'UTR regions, introns and the promoter of the gene product. These definitions generally refer to a single-stranded molecule, but in some embodiments will also encompass an additional strand that is partially, substantially or fully complementary to the single-stranded molecule. Thus, a nucleic acid can encompass a double-stranded molecule or a double-stranded molecule that comprises one or more complementary strand(s) or “complement(s)” of a particular sequence comprising a molecule. As used herein, a single stranded nucleic acid can be denoted by the prefix “ss”, a double stranded nucleic acid by the prefix “ds”, and a triple stranded nucleic acid by the prefix “ts.” The term “gene” refers to the segment of DNA involved in producing a polypeptide chain, it includes regions preceding and following the coding region as well as intervening sequences (introns) between individual coding segments (exons). A “promoter” is a region of a nucleic acid sequence at which initiation and rate of transcription are controlled. It can contain elements at which regulatory proteins and molecules can bind, such as RNA polymerase and other transcription factors, to initiate the specific transcription of a nucleic acid sequence. The term “enhancer” refers to a cisacting regulatory sequence involved in the transcriptional activation of a nucleic acid sequence. An enhancer can function in either orientation and can be upstream or downstream of the promoter. As used herein, the term “gene product(s)” is used to refer to include RNA transcribed from a gene, or a polypeptide encoded by a gene or translated from RNA.0157444.0804005

[0079] For the translocation of a gene into the nucleus of a target cell, a signal sequence may be conjugated to the MOI gene. Many nuclear signal sequences are known in the art. A nuclear membrane signal sequence or peptide is a sequence of amino acids generally of a length of about 10 to about 50 or more amino acid residues, many (typically about 55-60%) residues of which are hydrophobic such that they have a hydrophobic, lipid-soluble portion. Generally, a signal peptide is a peptide capable of penetrating through the cell membrane (e.g. the nuclear membrane) to allow the import and / or export of cellular proteins. Such a signal sequence may be naturally present in the gene or may be provided using many well known recombinant DNA techniques. Signal peptide sequences may be selected from the SIGPEP database (von Heijne, Protein Sequence Data Analysis 1 :4142 (1987); von Heijne and Abrahmsen, L., FEBS Letters 224:439-446 (1989)), the entire contents of both of which are incorporated herein by reference.

[0080] In some embodiments of the present disclosure, a method of transporting an MOI or effector agent to target cells includes conjugating a carrier peptide to an MOI or effector agent via a linker to form a complex, and providing the complex to the target cells or to a subject having the target cells. The carrier peptide, MOI (or effector agent), and the linker are as described herein.

[0081] Transport to the target cells may be determined by measuring the effect of the conjugated MOI or effector agent. In some embodiments, measuring the effect of the conjugated MOI or effector agent may include imaging the MOI or effector agent at the target. In additional embodiments, measuring the effect of the conjugated MOI or effector agent may include assaying the presence of the MOI or effector agent, by measuring an increase or decrease in an activity or level of MOI or effector agent. For example, if the MOI or effector agent is siRNA, the levels of the mRNA that correspond to the siRNA can be measured after providing a complex of the siRNA conjugated to a carrier peptide via a linker, in which an effect is indicative of the siRNA being delivered to the target.

[0082] In some embodiments, the method of transporting the MOI or effector agent occurs in a target cell selected from brain cells, spinal cord cells, glial cells, neurons, molecules and other cells of the central nervous system. In some embodiments, the method of transporting the MOI or effector agent to target cells includes providing the complex to a subject in which the target cells are within the blood brain barrier. In some embodiments, the subject having the target cells is a human. In some embodiments, the method of transporting the MOI or effector agent to a target that is found inside the BBB occurs in vitro.

[0083] In some embodiments of the method, the complex is formulated as a pharmaceutical composition. The terms “composition” and “pharmaceutical composition” are used interchangeably herein and refer to compositions or formulations that usually comprise an excipient, such as a pharmaceutically acceptable carrier, that is conventional in the art and that is suitable for administration to mammals, and for example, humans or0157444.0804005 human cells. Such compositions can be specifically formulated for administration via one or more of a number of routes, including but not limited to, oral, parenteral, intravenous, intraarterial, subcutaneous, intranasal, sublingual, intraspinal, intracerebroventricular, and the like. Cells administered a composition as disclosed herein can be part of a subject, for example for therapeutic, diagnostic, or prophylactic purposes. The cells can also be cultured, for example cells as part of an assay for screening potential pharmaceutical compositions, and the cells can be part of a transgenic animal for research purposes. In addition, compositions for topical (e.g., oral mucosa, respiratory mucosa) and / or oral administration can form solutions, suspensions, tablets, pills, capsules, sustained-release formulations, oral rinses, or powders, as known in the art are described herein. The compositions also can include stabilizers and preservatives. For examples of carriers, stabilizers and adjuvants, see University of the Sciences in Philadelphia (2005) Remington: The Science and Practice of Pharmacy with Facts and Comparisons, 21 st Ed.

[0084] Compositions as disclosed herein can be administered by any convenient route, including parenteral, enteral, mucosal, topical, e.g., subcutaneous, intravenous, topical, intramuscular, intraperitoneal, transdermal, rectal, vaginal, intranasal or intraocular. In one embodiment, the compositions as disclosed herein are not topically administered. In one embodiment, the delivery is by oral administration of the composition formulation. In one embodiment, the delivery is by intranasal administration of the composition, especially for use in therapy of the brain and related organs (e.g., meninges and spinal cord). Along these lines, intraocular administration is also possible. In another embodiment, the delivery means is by intravenous (i.v.) administration of the composition, which is especially advantageous when a longer-lasting i.v. formulation is desired. Suitable formulations can be found in Remington's Pharmaceutical Sciences, 16th and 18th Eds., Mack Publishing, Easton, Pa. (1980 and 1990), and Introduction to Pharmaceutical Dosage Forms, 4th Edition, Lea & Febiger, Philadelphia (1985), each of which is incorporated herein by reference.

[0085] The compositions as disclosed herein can be administered in prophylactically or therapeutically effective amounts. The targeted delivery compositions as disclosed herein can be administered along with a pharmaceutically acceptable carrier. A prophylactically or therapeutically effective amount means that amount necessary, at least partly, to attain the desired effect, or to delay the onset of, inhibit the progression of, or halt altogether, the onset or progression of the particular disease or disorder being treated. Such amounts will depend, of course, on the particular condition being treated, the severity of the condition and individual patient parameters including age, physical condition, size, weight and concurrent treatment. These factors are well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. It is preferred generally that a maximum dose be used, that is, the highest safe dose according to sound medical judgment. It will be understood by those of ordinary skill in the art, however, that a lower dose or tolerable dose can be administered for medical reasons, psychological reasons or for virtually any other reasons.0157444.0804005

[0086] The term “pharmaceutically acceptable carrier” means any pharmaceutically acceptable means to mix and / or deliver the targeted delivery composition to a subject. The term “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, involved in carrying or transporting the subject agents from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and is compatible with administration to a subject, for example a human. For the clinical use of the methods according to some embodiments of the present disclosure, targeted delivery compositions according to some embodiments are formulated into pharmaceutical compositions or pharmaceutical formulations for parenteral administration, e.g., intravenous; mucosal, e.g., intranasal; enteral, e.g., oral; topical, e.g., transdermal; ocular, e.g., via corneal scarification or other mode of administration. The pharmaceutical composition may contain a compound according to some embodiments in combination with one or more pharmaceutically acceptable ingredients. The carrier may be in the form of a solid, semi-solid or liquid diluent, cream or a capsule. Usually, the amount of active compounds is between 0.1-95% by weight of the preparation, for example, between 0.2-20% by weight in preparations for parenteral use and for example, between 1 and 50% by weight in preparations for oral administration.

[0087] The terms “parenteral administration” and “administered parenterally” as used herein refer to modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, sub capsular, subarachnoid, intraspinal, intracerebro spinal, and intrasternal injection and infusion. The phrases “systemic administration,” “administered systemically,” “peripheral administration” and “administered peripherally” as used herein mean the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the animal's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

[0088] As used herein, the terms “administering,” “introducing,” and “providing” are used interchangeably, and according to embodiments of the present disclosure, refer to the placement of the pharmaceutical composition including the carrier peptide, linker, and MOI or effector agent into a subject or in vitro culture by a method or route which results in at least partial localization of the MOIs or effector agents at a desired site (e.g. target cells and / or molecules). The MOIs or effector agents can be administered by any appropriate route which results in an effective treatment in the subject.

[0089] In the preparation of pharmaceutical formulations containing the targeted delivery compositions of the present disclosure in the form of dosage units for oral administration the compound selected can be mixed with solid, powdered ingredients, such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose derivatives,0157444.0804005 gelatin, or another suitable ingredient, as well as with disintegrating agents and lubricating agents such as magnesium stearate, calcium stearate, sodium stearyl fumarate and polyethylene glycol waxes. The mixture may then be processed into granules or pressed into tablets.

[0090] The following example is presented for illustrative purposes only, and does not limit the scope or content of the present disclosure.

[0091] Example of Receptor Mediated Transcytosis via carrier peptide. Western blot analysis (FIG. 3) was used to monitor brain protein levels wherein the carrier peptide and linker were coupled to dimeric forkhead associate (FHA) domain derived from KIF1 C coupled by a C-CC1 domain derived from KIF-1A.

[0092] The carrier peptide and linker were attached to the N-term or C-term forming a chimeric molecule of approximately 115 kDa. Protein was incubated with human neuronal cell line SH-SY5Y in cell culture media. After a 24hr incubation, cells were detached and washed. Total protein lysed was analyzed using western blots. Internalized total protein was detected with western blot and probed using anti- His antibodies.ILLUSTRATIVE COMBINATIONS

[0093] The following illustrative examples relate to various non-exhaustive ways in which the teachings herein may be combined or applied. It should be understood that the following examples are not intended to restrict the coverage of any claims that may be presented at any time in this application or in subsequent filings of this application. No disclaimer is intended. The following examples are being provided for nothing more than merely illustrative purposes. It is contemplated that the various teachings herein may be arranged and applied in numerous other ways. It is also contemplated that some variations may omit certain features referred to in the below examples. Therefore, none of the aspects or features referred to below should be deemed critical unless otherwise explicitly indicated as such at a later date by the inventors or by a successor in interest to the inventors. If any claims are presented in this application or in subsequent filings related to this application that include additional features beyond those referred to below, those additional features shall not be presumed to have been added for any reason relating to patentability.Example 1

[0094] A complex, comprising: a molecule of interest, biomolecule payload, or effector agent; a carrier peptide comprising a peptide represented by SEQ ID NO:1 or SEQ ID NO:34 or variants thereof: SEQ ID NO:1X1 -X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16SEQ ID NO:340157444.0804005X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16 in which:X1 is M or T;X2 is T or I;X3 is T or W;X4 is R or C;X5 is T or D;X6 is any amino acid, or Y, I, or G;X7 is F or Y;X8 is any amino acid, or T or C;X9 is any amino acid, or P, N, or S;X10 is any amino acid, or Y, T, or S;X12 is M or G;X14 is V or R;X15 is R, S, A, or I; andX16 is K, S, or D; and a polypeptide linker linking the carrier peptide to the molecule of interest, the biomolecule payload, or the effector agent.Example 2

[0095] The complex according to Example 1 , wherein the molecule of interest, biomolecule payload, or effector agent comprises a biological molecule.Example 3

[0096] The complex according to any preceding Example, wherein the molecule of interest, biomolecule payload, or effector agent comprises a biological molecule having a molecular weight of 10 kDa to 230 kDa.Example 4

[0097] The complex according to any preceding Example, wherein the molecule of interest, biomolecule payload, or effector agent comprises an entity selected from antibodies, antibody fragments, proteins, enzymatic protein domains, nucleic acid molecules, virus like particles, full native proteins, protein domains, partial protein domains with a binding affinity or a desired enzymatic activity.Example 5

[0098] The complex according to any preceding Example, wherein the molecule of interest, biomolecule payload, or effector agent comprises an entity selected from nucleic acid molecules such as RNAs and DNAs, aptamers, analogs, combinations of DNA and RNA, genes; peptides including antibody and antigen fragments; proteins, including antibodies, antibodies, engineered antibody like proteins; chemical molecules, viral0157444.0804005 particles, liposomes; endosomes; exosomes; nanoparticles, dendrimers; positron emission tomograph ligands; eukaryotic cells; prokaryotic cells; microspheres; nanogels; and bionanocapsules.Example 6

[0099] The complex according to any preceding Example, wherein the carrier peptide comprises a peptide selected from SEQ ID NOs:1 -7, 9, 10, 13-15, and 34-42.Example 7

[0100] The complex according to any preceding Example, wherein the carrier peptide comprises a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:13) in which:X3 is T or W;X4 is R or C;X5 is T or D;X6 is any amino acid;X7 is F or Y;X8 is any amino acid;X9 is any amino acid;X10 is any amino acid;X12 is M or G; andX14 is V or R.Example 8

[0101] The complex according to any preceding Example, wherein the carrier peptide comprises a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:14) in which:X3 is T or W;X4 is R or C;X5 is T or D;X6 is Y, G or I;X7 is F or Y;X8 is T or C;X9 is P, N, or S;X10 is Y, T, or S;X12 is M or G; andX14 is V.Example 90157444.0804005

[0102] The complex according to any preceding Example, wherein the carrier peptide comprises a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:15) in which:

[0103] X3 is T or W;

[0104] X4 is R or C;

[0105] X5 is T or D;

[0106] X6 is Y, G or i;

[0107] X7 is F or Y;

[0108] X8 is T;

[0109] X9 is P;

[0110] XI O is Y or T;

[0111] X12 is M or G; and

[0112] X14 is VExample 10

[0113] The complex according to any preceding Example, wherein the polypeptide linker comprises a peptide having 2 to 21 amino acids.Example 11

[0114] The complex according to any preceding Example, wherein the polypeptide linker comprises a peptide having 3 to 20 amino acids.Example 12

[0115] The complex according to any preceding Example, wherein the polypeptide linker comprises one or more amino acids selected from S, T, G, A, K, P, and E.Example 13

[0116] The complex according to any preceding Example, wherein the polypeptide linker comprises at least one P residue.Example 14

[0117] The complex according to any preceding Example, wherein the polypeptide linker comprises (X-Proline)n where X is any amino acid, and n is 1 to 10.Example 15

[0118] The complex according to any preceding Example, wherein the polypeptide linker comprises an alpha-helical linker.Example 160157444.0804005

[0119] The complex according to any preceding Example, wherein the polypeptide linker comprises a polypeptide selected from:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)PQHTGT (SEQ ID NO:47)STDKP (SEQ ID NO:48)SAQSA (SEQ ID NO:49)GGSGT (SEQ ID NQ:50)GSGSG (SEQ ID NO:51 )SGSGN (SEQ ID NO:52)SSTGK (SEQ ID NO:53)GTSDT (SEQ ID NO:54)SSSTG (SEQ ID NO:55)ASGST (SEQ ID NO:56)SKSTS (SEQ ID NO:57)TDGST (SEQ ID NO:58)PGTST (SEQ ID NO:59)SGAQQDSN (SEQ ID NQ:60)APKAPADG (SEQ ID NO:61 )PPASPKADAP (SEQ ID NO:62)SASTKGPS (SEQ ID NO:63)GQPKANPT (SEQ ID NO:64)AGATD (SEQ ID NO:65)NADAT (SEQ ID NO:66)SQSGD (SEQ ID NO:67)SAASSS (SEQ ID NO:68)GKSGK (SEQ ID NO:69)PGTST (SEQ ID NQ:70)PQTDP (SEQ ID NO:71 )PPASP (SEQ ID NO:72)KNTPP (SEQ ID NO:73)GSGSGS (SEQ ID NO:74)GSGSGS (SEQ ID NO:75)GSGSGSGS (SEQ ID NO:76)GSGSGSGSGS (SEQ ID NO:77)GSGSGSGSGSGS (SEQ ID NO:78)GSGSGSGSGSGSGS (SEQ ID NO:79)GSGSGSGSGSGSGSGS (SEQ ID NQ:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81 )GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82)GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)0157444.0804005GGSGGSGGSGGS (SEQ ID NO:85) GGSGGSGGSGGSGGS (SEQ ID NO:86) GGSGGSGGSGGSGGSGGS (SEQ ID NO:87) GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88) GGGS (SEQ ID NO:89) GGGSGGGS (SEQ ID NQ:90) GGGSGGGSGGGS (SEQ ID NO:91 ) GGGSGGGSGGGSGGGS (SEQ ID NO:92) GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93) EEA EEAEEA (SEQ ID NO:94) EEAEEAEEA (SEQ ID NO:95) EEAEEAEEAEEA (SEQ ID NO:96) EEAEEAEEAEEAEEA (SEQ ID NO:97) EEAEEAEEAEEAEEAEEA (SEQ ID NO:98) EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99) EEAS (SEQ ID NQ:100) EEASEEAS (SEQ ID NQ:101 ) EEASEEASEEAS (SEQ ID NQ:102) EEASEEASEEASEEAS (SEQ ID NO: 103) EEASEEASEEASEEASEEAS (SEQ ID NQ:104) XP where X is any amino acid XPXP where X is any amino acid XPXPXP where X is any amino acidXPXPXPXP where X is any amino acid (SEQ ID NO: 105) XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106) XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107) XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108) XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:109) XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110) XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 ) LE GGGGSGGGGSGGGGS (SEQ ID NO:112) GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113) GGGGGGGG (SEQ ID NO:114) GGGGGG (SEQ ID NO:115) EAAAKEAAAKEAAAK (SEQ ID NO:116) EAAAK (SEQ ID NO: 117)EAAAKEAAAK (SEQ ID NO: 118)AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID NQ:120) PAPAP (SEQ ID NO:121 ) AEAAAKEAAAKA (SEQ ID NO:122)0157444.0804005(AP)n where n is 5 to 17APAPAPAPAP (SEQ ID NO: 123)APAPAPAPAPAP (SEQ ID NO: 124)APAPAPAPAPAPAP (SEQ ID NO: 125)APAPAPAPAPAPAPAP (SEQ ID NO: 126)APAPAPAPAPAPAPAPAP (SEQ ID NO: 127) APAPAPAPAPAPAPAPAPAP (SEQ ID NO: 128) APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 )APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132) APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR j AETVFPDV (SEQ ID NO: 138)PLG | LWA (SEQ ID NO:139)RVL AEA (SEQ ID NO: 140)EDWCC 1 SMSY (SEQ ID NO:141 )GGIEGR 1 GS (SEQ ID NO:142)TRHRQPR 1 GWE (SEQ ID NO:143)AGNRVRR | SVG (SEQ ID NO:144)RRRRRRR j R J R (SEQ ID NO:145)GFLGj ((SEQ ID NO:146) valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) mc-ValCitPABC4-(4-acetylphenoxy) butanoic acid Hydrazone mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) m c-P E G -ca rbo n ate disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) wherein “me” is maleimidocaproyl.Example 17

[0120] The complex according to any preceding Example, wherein the polypeptide linker comprises a polypeptide selected from:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)0157444.0804005PQHTGT (SEQ ID NO:47)STDKP (SEQ ID NO:48)SAQSA (SEQ ID NO:49)GGSGT (SEQ ID NQ:50)GSGSG (SEQ ID NO:51 )SGSGN (SEQ ID NO:52)SSTGK (SEQ ID NO:53)GTSDT (SEQ ID NO:54)SSSTG (SEQ ID NO:55)ASGST (SEQ ID NO:56)SKSTS (SEQ ID NO:57)TDGST (SEQ ID NO:58)PGTST (SEQ ID NO:59)SGAQQDSN (SEQ ID NQ:60)APKAPADG (SEQ ID NO:61 )PPASPKADAP (SEQ ID NO:62)SASTKGPS (SEQ ID NO:63)GQPKANPT (SEQ ID NO:64)AGATD (SEQ ID NO:65)NADAT (SEQ ID NO:66)SQSGD (SEQ ID NO:67)SAASSS (SEQ ID NO:68)GKSGK (SEQ ID NO:69)PGTST (SEQ ID NQ:70)PQTDP (SEQ ID NO:71 )PPASP (SEQ ID NO:72)KNTPP (SEQ ID NO:73)GSGSGS (SEQ ID NO:74)GSGSGS (SEQ ID NO:75)GSGSGSGS (SEQ ID NO:76)GSGSGSGSGS (SEQ ID NO:77)GSGSGSGSGSGS (SEQ ID NO:78)GSGSGSGSGSGSGS (SEQ ID NO:79)GSGSGSGSGSGSGSGS (SEQ ID NQ:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81 )GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82) GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)GGSGGSGGSGGS (SEQ ID NO:85)GGSGGSGGSGGSGGS (SEQ ID NO:86)GGSGGSGGSGGSGGSGGS (SEQ ID NO:87)GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88)0157444.0804005GGGS (SEQ ID NO:89)GGGSGGGS (SEQ ID NO:90)GGGSGGGSGGGS (SEQ ID NO:91 )GGGSGGGSGGGSGGGS (SEQ ID NO:92)GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93)EEAEEAEEA (SEQ ID NO:94)EEAEEAEEA (SEQ ID NO:95)EEAEEAEEAEEA (SEQ ID NO:96)EEAEEAEEAEEAEEA (SEQ ID NO:97)EEAEEAEEAEEAEEAEEA (SEQ ID NO:98)EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99)EEAS (SEQ ID NQ:100)EEASEEAS (SEQ ID NQ:101 )EEASEEASEEAS (SEQ ID NQ:102)EEASEEASEEASEEAS (SEQ ID NO: 103)EEASEEASEEASEEASEEAS (SEQ ID NQ:104) XP where X is any amino acid XPXP where X is any amino acid XPXPXP where X is any amino acid XPXPXPXP where X is any amino acid (SEQ ID NO: 105) XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106) XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107) XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108) XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NQ:109) XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110) XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 )Example 18

[0121] The complex according to any of Examples 1-16, wherein the polypeptide linker comprises a polypeptide selected from:LEGGGGSGGGGSGGGGS (SEQ ID NO:112)GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113) GGGGGGGG (SEQ ID NO:114) GGGGGG (SEQ ID NO:115)EAAAKEAAAKEAAAK (SEQ ID NO:116)EAAAK (SEQ ID NO: 117)EAAAKEAAAK (SEQ ID NO: 118)AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID NQ:120) PAPAP (SEQ ID NO:121 )0157444.0804005AEAAAKEAAAKA (SEQ ID NO:122) (AP)n where n is 5 to 17 APAPAPAPAP (SEQ ID NO: 123) APAPAPAPAPAP (SEQ ID NO: 124) APAPAPAPAPAPAP (SEQ ID NO: 125) APAPAPAPAPAPAPAP (SEQ ID NO: 126) APAPAPAPAPAPAPAPAP (SEQ ID NO: 127) APAPAPAPAPAPAPAPAPAP (SEQ ID NO: 128) APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 ) APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132) APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR J AETVFPDV (SEQ ID NO: 138)PLG | LWA (SEQ ID NO:139)RVL AEA (SEQ ID NO: 140)EDWCC | SMSY (SEQ ID NO:141 )GGIEGR 1 GS (SEQ ID NO:142)TRHRQPR | GWE (SEQ ID NO:143)AGNRVRR 1 SVG (SEQ ID NO:144)RRRRRRR j R J R (SEQ ID NO:145)GFLGj ((SEQ ID NO:146) valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) mc-ValCitPABC4-(4-acetylphenoxy) butanoic acid Hydrazone mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) m c-P E G -ca rbo n ate disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) wherein “me” is maleimidocaproyl.Example 19

[0122] The complex according to any of Examples 1-12 and 15, wherein the polypeptide linker comprises a polypeptide represented by:(GS)n10157444.0804005(GGS)n2(GGGS (SEQ ID NO:43))n3(EEA)n1(EEAS (SEQ ID NO:44))n2 in which n1 is 1 -10, n2 is 1 -7, and / or n3 is 1-5.Example 20

[0123] The complex according to Example 19, wherein n1 is 1-6, n2 is 1-5, and / or n3 is 1-4.Example 21

[0124] The complex according to any preceding Example, wherein the complex comprises one carrier peptide and one polypeptide linker coupled to one side of the molecule of interest, biomolecule payload, or effector agent, for example at the N terminal side or the C terminal side of a protein or the chemically equivalent side of another nonprotein biomolecule.Example 22

[0125] The complex according to any preceding Example, wherein the complex comprises a first carrier peptide and a first polypeptide linker coupled to a first side of the molecule of interest, biomolecule payload, or effector agent, for example at the N terminal side of a protein or the chemically equivalent side of another non-protein biomolecule, and a second carrier peptide and a second polypeptide linker coupled to a second side of the molecule of interest, biomolecule payload, or effector agent, for example at the C terminal side of a protein or the chemically equivalent side of another non-protein biomolecule.Example 23

[0126] The complex according to any preceding Example, wherein the complex comprises two carrier peptides, each with a corresponding polypeptide linker, attached to one molecule of interest, biomolecule payload, or effector agent.Example 24

[0127] The complex according to Example 23, wherein the molecule of interest, biomolecule payload, or effector agent is an individual protein or protein fragment, and the N terminus and C terminus each have a respective carrier peptide and respective polypeptide linker.Example 250157444.0804005

[0128] The complex according to Example 24, wherein each of the respective carrier peptides have the same binding affinities to neuronal receptors.Example 26

[0129] The complex according to Example 24, wherein each of the respective carrier peptides have different binding affinities to neuronal receptors.Example 27

[0130] The complex according to any of Examples 1-20, the complex comprising at least two chimeric complexes, each with at least one respective carrier peptide and respective polypeptide linker, wherein one or more of the chimeric complexes forms a multimer.Example 28

[0131] The complex according to Example 27, wherein the multimer is a dimer, trimer, or tetramer.Example 29

[0132] The complex according to Example 27 or 28, wherein the multimerization region of the chimeric molecules is part of the molecule of interest, biomolecule payload, or effector agent, is at least a portion of the polypeptide linker(s), or is afforded by added chemical linkers.Example 30

[0133] The complex according to any of Examples 27-29, wherein the interaction between the chimeric molecules is covalent.Example 31

[0134] The complex according to any of Examples 27-29, wherein the interaction between the chimeric molecules is noncovalent.Example 32

[0135] The complex according to any of Examples 27-31 , wherein each of the respective carrier peptides have different binding affinities to neuronal receptors.Example 33

[0136] The complex according to any of Examples 27-31 , wherein each of the respective carrier peptides have the same binding affinities to neuronal receptors.0157444.0804005Example 34

[0137] A method of transporting a molecule of interest, biomolecule payload, or effector agent to target cells comprising providing the complex according to any preceding Example to the target cells or to a subject having the target cells.Example 25

[0138] A pharmaceutical composition, comprising: the complex according to any preceding Example; and one or more pharmaceutically acceptable excipients and / or carriers.Example 36

[0139] A method of transporting a molecule of interest, biomolecule payload, or effector agent to target cells comprising providing or administering the pharmaceutical composition according to Example 35 to the target cells or to a subject having the target cells.

[0140] While the subject matter of the present disclosure has been described in connection with certain embodiments, it is to be understood that the subject matter of the present disclosure is not limited to the disclosed embodiments, but, on the contrary, the present disclosure is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the present disclosure, as well as the appended claims, and equivalents thereof.

[0141] Also, as used herein, unless otherwise expressly specified, all numbers such as those expressing values, ranges, amounts or percentages may be read as if prefaced by the word “about,” even if the term does not expressly appear. As used herein, the terms “substantially”, “about”, and similar terms are used as terms of approximation and not as terms of degree, and are intended to account for the inherent deviations in measured or calculated values that would be recognized by those of ordinary skill in the art. Also, any numerical range recited herein is intended to include all sub-ranges of the same numerical precision subsumed within the recited range. For example, a range of “1.0 to 10.0” is intended to include all subranges between (and including) the recited minimum value of 1.0 and the recited maximum value of 10.0, that is, having a minimum value equal to or greater than 1.0 and a maximum value equal to or less than 10.0, such as, for example, 2.4 to 7.6. Any maximum numerical limitation recited herein is intended to include all lower numerical limitations subsumed therein and any minimum numerical limitation recited in this specification is intended to include all higher numerical limitations subsumed therein. Accordingly, Applicant reserves the right to amend this specification, including the claims, to expressly recite any sub-range subsumed within the ranges expressly recited herein. As used herein, the terms “combination thereof” and “combinations thereof” may refer to a chemical combination (e.g., an alloy or chemical compound), a mixture, or a solution with multiple solutes.0157444.0804005

[0142] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the present disclosure. Plural encompasses singular and vice versa. As used herein, the singular forms “a” and “an” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprises”, “comprising”, “includes”, and “including,” when used in this specification, specify the presence of the stated features, integers, acts, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, acts, operations, elements, components, and / or groups thereof. As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items. Expressions such as “at least one of”, when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

Claims

0157444.0804005WHAT IS CLAIMED IS:

1. A complex, comprising: a molecule of interest, biomolecule payload, or effector agent; a carrier peptide comprising a peptide represented by SEQ ID NO:1 or SEQ ID NO:34 or variants thereof:SEQ ID NO:1Xi -X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16SEQ ID NO:34X2-X3-X4-X5-X6-X7-X8-X9-X10-R-X12-K-X14-X15-X16 in which:Xi is M or T;X2 is T or I;X3is T or W;X4 is R or C;Xs is T or D;Xe is any amino acid, or Y, I, or G;X7 is F or Y;Xs is any amino acid, or T or C;X9 is any amino acid, or P, N, or S;X10 is any amino acid, or Y, T, or S;X12 is M or G;X14 is V or R;X15 is R, S, A, or I; andX16 is K, S, or D; and a polypeptide linker linking the carrier peptide to the molecule of interest, the biomolecule payload, or the effector agent.

2. The complex according to claim 1 , wherein the molecule of interest, biomolecule payload, or effector agent comprises a biological molecule.

3. The complex according to any preceding claim, wherein the molecule of interest, biomolecule payload, or effector agent comprises a biological molecule having a molecular weight of 10 kDa to 230 kDa.0157444.08040054. The complex according to any preceding claim, wherein the molecule of interest, biomolecule payload, or effector agent comprises an entity selected from antibodies, antibody fragments, proteins, enzymatic protein domains, nucleic acid molecules, virus like particles, full native proteins, protein domains, partial protein domains with a binding affinity or a desired enzymatic activity.

5. The complex according to any preceding claim, wherein the molecule of interest, biomolecule payload, or effector agent comprises an entity selected from nucleic acid molecules such as RNAs and DNAs, aptamers, analogs, combinations of DNA and RNA, genes; peptides including antibody and antigen fragments; proteins, including antibodies, antibodies, engineered antibody like proteins; chemical molecules, viral particles, liposomes; endosomes; exosomes; nanoparticles, dendrimers; positron emission tomograph ligands; eukaryotic cells; prokaryotic cells; microspheres; nanogels; and bionanocapsules.

6. The complex according to any preceding claim, wherein the carrier peptide comprises a peptide selected from SEQ ID NOs:1 -7, 9, 10, 13-15, and 34-42.

7. The complex according to any preceding claim, wherein the carrier peptide comprises a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:13) in which:X3is T or W;X4 is R or C;Xs is T or D;Xe is any amino acid;X7 is F or Y;Xs is any amino acid;X9 is any amino acid;X10 is any amino acid;X12 is M or G; andX14 is V or R.0157444.08040058. The complex according to any preceding claim, wherein the carrier peptide comprises a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:14) in which:X3is T or W;X4 is R or C;Xs is T or D;X6is Y, G or I;X7is F or Y;Xs is T or C;X9 is P, N, or S;X10 is Y, T, or S;X12 is M or G; and X14 is V.

9. The complex according to any preceding claim, wherein the carrier peptide comprises a peptide represented by X3-X4-X5-X6-X7-X8-X9-X10-R11-X12-K13-X14 (SEQ ID NO:15) in which:X3is T or W;X4 is R or C;Xs is T or D;X6is Y, G or I;X7is F or Y;X8is T;X9is P;X10 is Y or T;X12 is M or G; and X14 is V.

10. The complex according to any preceding claim, wherein the polypeptide linker comprises a peptide having 2 to 21 amino acids.11 . The complex according to any preceding claim, wherein the polypeptide linker comprises a peptide having 3 to 20 amino acids.0157444.080400512. The complex according to any preceding claim, wherein the polypeptide linker comprises one or more amino acids selected from S, T, G, A, K, P, and E.

13. The complex according to any preceding claim, wherein the polypeptide linker comprises at least one P residue.

14. The complex according to any preceding claim, wherein the polypeptide linker comprises (X-Proline)n where X is any amino acid, and n is 1 to 10.

15. The complex according to any preceding claim, wherein the polypeptide linker comprises an alpha-helical linker.

16. The complex according to any preceding claim, wherein the polypeptide linker comprises a polypeptide selected from:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)PQHTGT (SEQ ID NO:47)STDKP (SEQ ID NO:48)SAQSA (SEQ ID NO:49)GGSGT (SEQ ID NQ:50)GSGSG (SEQ ID NO:51 )SGSGN (SEQ ID NO:52)SSTGK (SEQ ID NO:53)GTSDT (SEQ ID NO:54)SSSTG (SEQ ID NO:55)ASGST (SEQ ID NO:56)SKSTS (SEQ ID NO:57)TDGST (SEQ ID NO:58)PGTST (SEQ ID NO:59)SGAQQDSN (SEQ ID NQ:60)APKAPADG (SEQ ID NO:61 )PPASPKADAP (SEQ ID NO:62)0157444.0804005SASTKGPS (SEQ ID NO:63)GQPKANPT (SEQ ID NO:64)AGATD (SEQ ID NO:65)NADAT (SEQ ID NO:66)SQSGD (SEQ ID NO:67)SAASSS (SEQ ID NO:68)GKSGK (SEQ ID NO:69)PGTST (SEQ ID NQ:70)PQTDP (SEQ ID NO:71 )PPASP (SEQ ID NO:72)KNTPP (SEQ ID NO:73)GSGSGS (SEQ ID NO:74)GSGSGS (SEQ ID NO:75)GSGSGSGS (SEQ ID NO:76)GSGSGSGSGS (SEQ ID NO:77)GSGSGSGSGSGS (SEQ ID NO:78)GSGSGSGSGSGSGS (SEQ ID NO:79)GSGSGSGSGSGSGSGS (SEQ ID NQ:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81 ) GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82) GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)GGSGGSGGSGGS (SEQ ID NO:85)GGSGGSGGSGGSGGS (SEQ ID NO:86)GGSGGSGGSGGSGGSGGS (SEQ ID NO:87)GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88) GGGS (SEQ ID NO:89)GGGSGGGS (SEQ ID NQ:90)GGGSGGGSGGGS (SEQ ID NO:91 )GGGSGGGSGGGSGGGS (SEQ ID NO:92) GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93)0157444.0804005EEAEEAEEA (SEQ ID NO:94)EEAEEAEEA (SEQ ID NO:95)EEAEEAEEAEEA (SEQ ID NO:96)EEAEEAEEAEEAEEA (SEQ ID NO:97)EEAEEAEEAEEAEEAEEA (SEQ ID NO:98)EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99) EEAS (SEQ ID NQ:100) EEASEEAS (SEQ ID NQ:101 ) EEASEEASEEAS (SEQ ID NQ:102) EEASEEASEEASEEAS (SEQ ID NO: 103) EEASEEASEEASEEASEEAS (SEQ ID NQ:104) XP where X is any amino acid XPXP where X is any amino acid XPXPXP where X is any amino acid XPXPXPXP where X is any amino acid (SEQ ID NO: 105) XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106) XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107) XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108) XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NQ:109) XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110) XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 ) LEGGGGSGGGGSGGGGS (SEQ ID NO:112) GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113) GGGGGGGG (SEQ ID NO:114) GGGGGG (SEQ ID NO:115) EAAAKEAAAKEAAAK (SEQ ID NO:116) EAAAK (SEQ ID NO: 117) EAAAKEAAAK (SEQ ID NO: 118) AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID N PAPAP (SEQ ID N AEAAAKEAAAKA ( (AP)n where n is 5 t APAPAPAPAP (SE0157444.0804005APAPAPAPAPAP (SEQ ID NO: 124)APAPAPAPAPAPAP (SEQ ID NO: 125)APAPAPAPAPAPAPAP (SEQ ID NO: 126)APAPAPAPAPAPAPAPAP (SEQ ID NO: 127) APAPAPAPAPAPAPAPAPAP (SEQ ID NO: 128) APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 ) APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132) APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR J AETVFPDV (SEQ ID NO: 138)PLG 1 LWA (SEQ ID NO:139)RVL AEA (SEQ ID NO: 140)EDWCC 1 SMSY (SEQ ID NO:141 )GGIEGR 1 GS (SEQ ID NO:142)TRHRQPR | GWE (SEQ ID NO:143)AGNRVRR 1 SVG (SEQ ID NO:144)RRRRRRR J R J R (SEQ ID NO:145)GFLG1 ((SEQ ID NO:146) valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) mc-ValCitPABC 4-(4-acetylphenoxy) butanoic acid Hydrazone mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) m c-P E G -ca rbo n ate disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) wherein “me” is maleimidocaproyl.

17. The complex according to any preceding claim, wherein the polypeptide linker comprises a polypeptide selected from:SKSAD (SEQ ID NO:45)TTAGN (SEQ ID NO:46)PQHTGT (SEQ ID NO:47)0157444.0804005STDKP (SEQ ID NO:48) SAQSA (SEQ ID NO:49) GGSGT (SEQ ID NQ:50) GSGSG (SEQ ID NO:51 ) SGSGN (SEQ ID NO:52) SSTGK (SEQ ID NO:53) GTSDT (SEQ ID NO:54) SSSTG (SEQ ID NO:55) ASGST (SEQ ID NO:56) SKSTS (SEQ ID NO:57) TDGST (SEQ ID NO:58) PGTST (SEQ ID NO:59) SGAQQDSN (SEQ ID NQ:60) APKAPADG (SEQ ID NO:61 ) PPASPKADAP (SEQ ID NO:62) SASTKGPS (SEQ ID NO:63) GQPKANPT (SEQ ID NO:64) AGATD (SEQ ID NO:65) NADAT (SEQ ID NO:66) SQSGD (SEQ ID NO:67) SAASSS (SEQ ID NO:68) GKSGK (SEQ ID NO:69) PGTST (SEQ ID NQ:70) PQTDP (SEQ ID NO:71 ) PPASP (SEQ ID NO:72) KNTPP (SEQ ID NO:73) GSGSGS (SEQ ID NO:74) GSGSGS (SEQ ID NO:75) GSGSGSGS (SEQ ID NO:76) GSGSGSGSGS (SEQ ID NO:77) GSGSGSGSGSGS (SEQ ID NO:78) GSGSGSGSGSGSGS (SEQ ID NO:79)0157444.0804005GSGSGSGSGSGSGSGS (SEQ ID NO:80)GSGSGSGSGSGSGSGSGS (SEQ ID NO:81 )GSGSGSGSGSGSGSGSGSGS (SEQ ID NO:82)GGSGGSGGS (SEQ ID NO:83)GGSGGSGGS (SEQ ID NO:84)GGSGGSGGSGGS (SEQ ID NO:85)GGSGGSGGSGGSGGS (SEQ ID NO:86)GGSGGSGGSGGSGGSGGS (SEQ ID NO:87)GGSGGSGGSGGSGGSGGSGGS (SEQ ID NO:88)GGGS (SEQ ID NO:89)GGGSGGGS (SEQ ID NQ:90)GGGSGGGSGGGS (SEQ ID NO:91 )GGGSGGGSGGGSGGGS (SEQ ID NO:92)GGGSGGGSGGGSGGGSGGGS (SEQ ID NO:93)EEAEEAEEA (SEQ ID NO:94)EEAEEAEEA (SEQ ID NO:95)EEAEEAEEAEEA (SEQ ID NO:96)EEAEEAEEAEEAEEA (SEQ ID NO:97)EEAEEAEEAEEAEEAEEA (SEQ ID NO:98)EEAEEAEEAEEAEEAEEAEEA (SEQ ID NO:99)EEAS (SEQ ID NQ:100)EEASEEAS (SEQ ID NQ:101 )EEASEEASEEAS (SEQ ID NQ:102)EEASEEASEEASEEAS (SEQ ID NO: 103)EEASEEASEEASEEASEEAS (SEQ ID NQ:104)XP where X is any amino acidXPXP where X is any amino acidXPXPXP where X is any amino acidXPXPXPXP where X is any amino acid (SEQ ID NO: 105)XPXPXPXPXP where X is any amino acid (SEQ ID NO: 106)XPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 107)0157444.0804005XPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO: 108)XPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:109)XPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:110)XPXPXPXPXPXPXPXPXPXP where X is any amino acid (SEQ ID NO:111 )18. The complex according to any of claims 1 -16, wherein the polypeptide linker comprises a polypeptide selected from:LEGGGGSGGGGSGGGGS (SEQ ID NO:112)GGGGSGGGGSGGGGSGGGGS (SEQ ID NO:113) GGGGGGGG (SEQ ID NO:114) GGGGGG (SEQ ID NO:115)EAAAKEAAAKEAAAK (SEQ ID NO:116)EAAAK (SEQ ID NO: 117)EAAAKEAAAK (SEQ ID NO: 118)AEAAAKEAAAKEAAAKEAAAKALEAEAAAKEAAAKEAAAKEAAAKA (SEQ ID NO:119)GGGGS (SEQ ID NO:120)PAPAP (SEQ ID NO:121 )AEAAAKEAAAKA (SEQ ID NO:122)(AP)nwhere n is 5 to 17APAPAPAPAP (SEQ ID NO: 123)APAPAPAPAPAP (SEQ ID NO: 124)APAPAPAPAPAPAP (SEQ ID NO: 125)APAPAPAPAPAPAPAP (SEQ ID NO: 126)APAPAPAPAPAPAPAPAP (SEQ ID NO: 127)APAPAPAPAPAPAPAPAPAP (SEQ ID NO: 128) APAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 129) APAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 130) APAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 131 ) APAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 132) APAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 133) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 134) APAPAPAPAPAPAPAPAPAPAPAPAPAPAPAPAP (SEQ ID NO: 135) MCC mc-MMAF (MMAF is SEQ ID NO:136)RKKRR (SEQ ID NO:137)RXK (where X is any amino acid)RRDisulfideVSQTSKLTR J AETVFPDV (SEQ ID NO: 138)0157444.0804005PLG | LWA (SEQ ID NO:139)RVL AEA (SEQ ID NO: 140)EDWCC | SMSY (SEQ ID NO:141 )GGIEGR 1 GS (SEQ ID NO:142)TRHRQPR | GWE (SEQ ID NO:143)AGNRVRR 1 SVG (SEQ ID NO:144)RRRRRRR j R j R (SEQ ID NO:145)GFLGj ((SEQ ID NO:146) valine-citrulline-p-aminobenzyl carbamate (ValCitPABC) mc-ValCitPABC 4-(4-acetylphenoxy) butanoic acid Hydrazone mc-GGFG aminomethoxy (GGFG is SEQ ID NO:147) mc-PEG-carbonate disulfide-containing cleavable linker sulfo-SPDB (SPDB is SEQ ID NO: 148) wherein “me” is maleimidocaproyl.

19. The complex according to any of claims 1 -12 and 15, wherein the polypeptide linker comprises a polypeptide represented by:(GS)n1(GGS)n2(GGGS (SEQ ID NO:43))n3(EEA)n1(EEAS (SEQ ID NO:44))n2in which m is 1 -10, n2is 1-7, and / or n3is 1-5.

20. The complex according to claim 19, wherein m is 1 -6, n2is 1 -5, and / or n3is 1 -4.21 . The complex according to any preceding claim, wherein the complex comprises one carrier peptide and one polypeptide linker coupled to one side of the molecule of interest, biomolecule payload, or effector agent, for example at the N terminal side or the C terminal side of a protein or the chemically equivalent side of another non-protein biomolecule.

22. The complex according to any preceding claim, wherein the complex comprises a first carrier peptide and a first polypeptide linker coupled to a first side of the molecule of interest, biomolecule payload, or effector agent, for example at the N terminal side of a0157444.0804005 protein or the chemically equivalent side of another non-protein biomolecule, and a second carrier peptide and a second polypeptide linker coupled to a second side of the molecule of interest, biomolecule payload, or effector agent, for example at the C terminal side of a protein or the chemically equivalent side of another non-protein biomolecule.

23. The complex according to any preceding claim, wherein the complex comprises two carrier peptides, each with a corresponding polypeptide linker, attached to one molecule of interest, biomolecule payload, or effector agent.

24. The complex according to claim 23, wherein the molecule of interest, biomolecule payload, or effector agent is an individual protein or protein fragment, and the N terminus and C terminus each have a respective carrier peptide and respective polypeptide linker.

25. The complex according to claim 24, wherein each of the respective carrier peptides have the same binding affinities to neuronal receptors.

26. The complex according to claim 24, wherein each of the respective carrier peptides have different binding affinities to neuronal receptors.

27. The complex according to any of claims 1 -20, the complex comprising at least two chimeric complexes, each with at least one respective carrier peptide and respective polypeptide linker, wherein one or more of the chimeric complexes forms a multimer.

28. The complex according to claim 27, wherein the multimer is a dimer, trimer, or tetramer.

29. The complex according to claim 27 or 28, wherein the multimerization region of the chimeric molecules is part of the molecule of interest, biomolecule payload, or effector agent, is at least a portion of the polypeptide linker(s), or is afforded by added chemical linkers.

30. The complex according to any of claims 27-29, wherein the interaction between the chimeric molecules is covalent.0157444.080400531 . The complex according to any of claims 27-29, wherein the interaction between the chimeric molecules is noncovalent.

32. The complex according to any of claims 27-31 , wherein each of the respective carrier peptides have different binding affinities to neuronal receptors.

33. The complex according to any of claims 27-31 , wherein each of the respective carrier peptides have the same binding affinities to neuronal receptors.

34. A method of transporting a molecule of interest, biomolecule payload, or effector agent to target cells comprising providing the complex according to any preceding claim to the target cells or to a subject having the target cells.

35. A pharmaceutical composition, comprising: the complex according to any preceding claim; and one or more pharmaceutically acceptable excipients and / or carriers.

36. A method of transporting a molecule of interest, biomolecule payload, or effector agent to target cells comprising providing or administering the pharmaceutical composition according to claim 35 to the target cells or to a subject having the target cells.

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