Modified pigment epithelium-derived factor peptides and methods of use

Abstract

Provided herein are compositions and methods for treating a retinal disease or disorder. In some examples, the compositions include a pigment epithelium-derived factor (PEDF) peptide including one or more of: an N-terminal modification, one or more amino acid substitutions, deletion of 1-4 amino acids, replacement of one or more amino acids with a non-alpha-amino acid linker, a C-terminal modification, or any combination of two or more thereof. Methods for treating a retinal disease or disorder include administering a provided peptide to an eye of a subject with the retinal disease or disorder, such as retinal degeneration or macular degeneration.

Description

[0001] MODIFIED PIGMENT EPITHELIUM-DERIVED FACTOR PEPTIDES AND METHODS

[0002] OF USE

[0003] CROSS REFERENCE TO RELATED APPLICATIONS

[0004] This application claims the benefit of U.S. Provisional Application No. 63 / 604,026, filed November 29, 2023, which is incorporated by reference herein in its entirety.

[0005] FIELD

[0006] This disclosure relates to modified pigment epithelium-derived factor peptides and methods of their use, particularly for treating retinal degeneration.

[0007] ACKNOWLEDGMENT OF GOVERNMENT SUPPORT

[0008] This invention was made with Government support under project numbers ZIA EY000438 and ZIA EY000306 by the National Institutes of Health, National Eye Institute. The Government has certain rights in the invention.

[0009] SEQUENCE LISTING INCORPORATION

[0010] The Sequence Listing is submitted as an XML file in the form of the file named “Sequence.xml” (~109,276 bytes), which was created on October 30, 2024, which is incorporated by reference herein.

[0011] BACKGROUND

[0012] Inherited Retinal Degenerations (IRD), such as Retinitis Pigmentosa (RP), are characterized by a progressive loss of visual acuity caused by degeneration of photoreceptor cells. RP can arise from mutations in more than 90 genes and the phenotype of the disease is highly variable. Due to this high heterogeneity, therapeutic approaches targeting specific genes generally benefit few patients, while for most forms of RP, few or no treatment options are available. Thus, there remains a need to identify new and more effective treatments for RP and other retinal diseases of disorders.

[0013] SUMMARY

[0014] Provided herein are compositions and methods that utilize pigment epithelium-derived factor (PEDF) peptides to inhibit or delay photoreceptor cell loss and blindness. Thus, provided herein are compositions and methods for treating a retinal disease or disorder, such as a retinal degeneration or macular degeneration.

[0015] Provided herein are pigment epithelium-derived factor (PEDF) peptides including the amino acid of SEQ ID NO: 1 with one or more modifications, wherein the one or more modifications are selected from: a) an N-terminal modification; b) one or more amino acid substitutions; c) deletion of 1-4 amino acids from the N-terminus and / or C-terminus of the peptide; d) replacement of one or more amino acids with a non-alpha-amino acid linker; e) a C-terminal modification; and f) any combination of two or more thereof.

[0016] In some examples, the peptide includes 8-19 amino acids (such as 8-12 or 13-17 amino acids).

[0017] In some examples, the N-terminal modification of the PEDF peptide includes 6- aminohexanoic acid (AHA), aminopentanoic acid (APA), aminobutanoic acid (ABA), beta-alanine, or N-acetyl, or is derived from AHA, APA, ABA, or beta-alanine.

[0018] In some examples, the PEDF peptide includes one or more amino acid substitutions at one or more positions corresponding to amino acid position 5, 6, or 8 of SEQ ID NO: 1. In some examples, the one or more amino acid substitutions includes threonine, aspartic acid, diaminopropionic acid (DAP), or CFs-threonine at the position corresponding to amino acid position 5 of SEQ ID NO: 1; norleucine, leucine, valine, or N-methylated isoleucine at the position corresponding to amino acid position 6 of SEQ ID NO: 1; and / or alanine at the position corresponding to amino acid position 8 of SEQ ID NO: 1.

[0019] In some examples, the PEDF peptide includes deletion of 1-4 amino acids from the N- terminus of the peptide, deletion of 1-4 amino acids from the C-terminus of the peptide, or both.

[0020] In additional examples, the PEDF peptide includes a non-alpha-amino acid linker such as 6- aminohexanoic acid (AHA), 5 -aminopentanoic acid (APA), 8-amino-3,6,dioxaoctanoic acid (AOC), 4’-amino-[l,l’-biphenyl]-4-carboxylate (4’ABPC), or 4-(4-(aminomethyl)phenoxy)benzoic acid (4’AMPC), 8-aminooctanoic acid (AOA), or is derived from AHA, APA, AOC, 4’ABPC, 4’AMPC, or AOA. In some examples, the peptide includes substitution of the amino acid corresponding to position 8, 9, or 10 of SEQ ID NO: 1 with the non- alpha-amino acid linker. In some examples, the PEDF peptide further includes deletion of 1-3 amino acids following the position of substitution with the non-alpha-amino acid linker.

[0021] In some examples, the PEDF peptide includes a C-terminal modification, such as C- terminal amidation, benzylamine- NH2, NH2-ethyl, or NH2CH2CH2OH. In further examples, the PEDF peptide increases phospholipase Ai activity of PEDF receptor rhuPEDF-R( 1-288), increases phospholipase A2 activity of PEDF receptor rhuPEDF-R(l- 288), or both, compared to a control.

[0022] In some examples, the PEDF peptide includes or consists of the amino acid sequence of any one of SEQ ID NOs: 3-54 or 61-64.

[0023] Also provided are compositions including one or more of the disclosed PEDF peptides and a pharmaceutically acceptable carrier. In some examples, the pharmaceutically acceptable carrier is water or buffered saline, for example, Hank’s balanced salt solution. In some examples, the composition is formulated as an eyedrop or other topical formulation. In other examples, the composition is formulated for injection. In some examples, the composition includes about 1 mg / ml of the peptide.

[0024] Methods of treating a retinal disease or disorder are provided, including administering a composition including one or more of the disclosed PEDF peptides to an eye of a subject with the retinal disease or disorder. In some examples, the composition is administered topically to the eye of the subject. In other examples, the composition is administered to the eye of the subject by injection. In some examples, the composition may be administered to the eye of the subject twice daily, once daily, every other day, or every three days. In some examples, the retinal disease or disorder is retinal degeneration (such as retinitis pigmentosa, Leber congenital amaurosis, or conerod dystrophy), macular degeneration (such as age-related macular degeneration, Stargardt-like macular degeneration, vitelliform macular dystrophy (Best disease), bull’s eye maculopathy, or other maculopathy), or glaucoma.

[0025] Methods of treating or inhibiting cancer are also provided, including administering a composition including one or more of the disclosed PEDF peptides to a subject with cancer. In some examples, the subject has ovarian cancer, liver cancer, pancreatic cancer, or melanoma.

[0026] Methods of treating or inhibiting a central nervous system (CNS) disease or disorder are also provided, the methods including a composition including one or more of the disclosed PEDF peptides to a subject with the CNS disease or disorder. In some examples, the CNS disease or disorder is a disease or disorder that includes cell death in the CNS. In particular examples, the CNS disease or disorder is amyotrophic lateral sclerosis, Huntington’s disease, Alzheimer’s disease, Parkinson’s disease, human immunodeficiency virus-induced encephalitis, or CNS trauma.

[0027] The foregoing and other features of this disclosure will become more apparent from the following detailed description of several aspects which proceeds with reference to the accompanying figures. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] FIG. 1 is a schematic showing a comparison of peptides SEQ ID NOs: 12, 13, 16, and 19 to wild type PEDF 17mer (SEQ ID NO: 1). The plot is adapted from Kenealey et al. (J. Biol. Chem. 90(42):25241-25253, 2015) showing binding of peptides from an alanine scan series to PEDF receptor PEDF-R-P1 peptide.

[0029] FIGS. 2A and 2B show the effect of PIO (Pep. 10 (SEQ ID NO: 12)) and Pl 1 (Pep. 11 (SEQ ID NO: 13)) peptide administration on PR cell death in both rdlO (FIG. 2A) and rdlO / Serpinfl^' (FIG. 2B) mice. For all experiments shown, three retinas per group were evaluated and each data point corresponds to the average ± SEM of fluorescence relative to HBSS by unpaired t-test. ***p < 0.0001, ****p < 0.00001.

[0030] FIG. 3 shows the effect of P14 (Pep. 14 (SEQ ID NO: 16)) peptide administration on PR cell death in both rdlO and rdlO / Serpinfl ^ mice. For all experiments shown, three retinas per group were evaluated and each data point corresponds to the average ± SEM of fluorescence relative to HBSS by unpaired t-test. **p < 0.001, ****p < 0.00001.

[0031] FIGS. 4A and 4B show the effect of daily eye drops of P10 (SEQ ID NO: 12), Pl 1 (SEQ ID NO: 13), and P14 (SEQ ID NO: 16) peptide administration on PR cell death in both rdlO (FIG. 4A) and rdlO / Serpinfl ^ (FIG. 4B) mice at day P21. An average of three retinas per group were evaluated and each data point corresponds to the average of fluorescence relative to HBSS. ***p < 0.0001

[0032] FIGS. 5 A and 5B show histological evaluation of retinas in mice treated with P10 peptide (SEQ ID NO: 12). Hematoxylin and eosin staining and measurement of outer nuclear layer (ONL) thickness are shown for rdlO (FIG. 5A) and rdlO / Serpinfl ' (FIG. 5B) mice. For all histology shown, three retinas per group were evaluated and each data point corresponds to the average + SD per location relative to the optic nerve per genotype by unpaired t test. ***p < 0.0001, ****p < 0.00001

[0033] FIGS. 6A-6E show effects of peptides Pl (SEQ ID NO: 3), P18 (SEQ ID NO: 20), P19 (SEQ ID NO: 21), and P5 (SEQ ID NO: 7) in rdlO mice. FIG. 6A is a schematic diagram showing the experimental timeline. FIG. 6B is a series of fluorescence fundoscopy images showing PS externalization of retinas from rdlO mice treated with eyedrops containing the indicated peptide at 1 mg / ml. The contralateral eye received vehicle control (H2O). FIG. 6C is a series of graphs showing quantification of fluorescence PS externalization. Mean intensity of fluorescence of eyes treated with H2O was subtracted from the fluorescence of eyes treated with peptide and plotted. N=3 and each data point represents average ± SD of fluorescence relative to water by unpaired t- test. ** p< 0.001, *** p<0.0001. FIG. 6D shows images of retina sections of rdlO mice at P21 stained with hematoxylin and eosin. OS, outer segment; ONL, outer nuclear layer; INL, inner nuclear layer. FIG. 6E is a spider plot of the thickness of the ONL as a function of distance from the optic nerve (ON). For all histology shown, five retinas per group and two sections per retina were evaluated and each data point corresponds to the average ± SD of ONL thickness per location by unpaired t-test. **p < 0.001, ***p < 0.0001, ****p < 0.00001.

[0034] FIG. 7 is a schematic diagram showing an expression vector of recombinant human rhuPEDF-R(l-288) in a pE-SUMOstar vector. Vector not drawn to scale. Xbal overhang is generated by Bsal.

[0035] FIGS. 8A-8C show production and purification of rhuPEDF-R(l-288). FIG. 8A illustrates exemplary purification steps of rhuPEDF-R(l-288). FIG. 8B shows Coomassie blue stained SDS- polyacrylamide gel analysis of purification fractions, confirming high purity of the expected protein, which migrates at the expected ~51 kDa. FIG. 8C is a Western blot of the purified rhuPEDF-R(l-288) protein, recognized by three specific antibodies as indicated.

[0036] FIGS. 9A-9C show characterization of PLA enzymatic activity of rhuPEDF-R(l-288). FIG. 9A shows that the recombinant protein displays PLA2 activity, as well as PLAi activity. FIGS. 9B and 9C are controls showing that the substrate used in FIG. 10A for PLAi activity (lecitase ultra) does not hydrolyze the PLA2 substrate (FIG. 10B) and that the substrate used in FIG. 10A for PLA2 activity (honey bee venom) does not hydrolyze the PLAi substrate (FIG. 10C).

[0037] FIG. 10 is a plot of PLA2 activity of rhuPEDF-R(l-288) (normalized fluorescence intensity at 515 nm) fold changes when in the presence of different concentrations of PEDF 17-mer (SEQ ID NO: 1), H105A (SEQ ID NO: 2), and R99A (SEQ ID NO: 57) peptides relative to activity without peptide. Enzymatic reactions were performed with rhuPEDF-R(l-288), liposome substrate and without or with peptide, and incubated for 10 minutes at room temperature. Fluorescence was measured with excitation at 460 nm on a Spectra Max® 3. The final concentration of rhuPEDF- R(l-288) was 60 nM and reaction volume was 100 pl. The reaction was performed in triplicate for each peptide concentration.

[0038] FIG. 11 is a plot of PLA2 activity of rhuPEDF-R(l-288) fold changes over reaction without peptide (dotted line) in the presence of PEDF-derived modified peptides (250 pM). Each bar represents the average of three independent experiments, each of which were performed in triplicate for each peptide. The final concentration of rhuPEDF-R (1-288) was 60 nM and the reaction volume was 100 pl.

[0039] FIG. 12 is a plot of PLA2 activity of rhuPEDF-R(l-288) fold changes in the presence of different concentrations of H105A, Pl, P18, P19, and P28 peptides. The final concentration of PEDF-R (1-288) was 60 nM. The plot represents an average of two independent experiments with reactions performed in triplicate for each concentration.

[0040] FIGS. 13A and 13B are plots of PLA2(FIG. 13A) and PLAi (FIG. 13B) activity (normalized emission intensity) versus concentration of rhuPEDF(l-288). Reaction mixtures with rhuPEDF-R(l-288) were incubated at room temperature for 10 min (PLA2) or 30 min (PLAi) with liposome substrate. Fluorescence was measured with excitation at 460 nm on a Spectra Max® 3 plate reader. The final reaction volume was 100 pl. Each concentration of rhuPEDF-R(l-288) was tested in triplicate.

[0041] FIG. 14 is a plot of PLAi activity of rhuPEDF-R(l-288) fold changes over reaction without peptide (dotted line) in the presence of PEDF derived modified peptides (250 pM). Each bar represents the average of three independent experiments, each of which were performed in triplicate for each peptide. The final concentration of rhuPEDF-R( 1-288) was 60 nM and the reaction volume was 100 pl.

[0042] FIG. 15 is a plot of PLAi activity of rhuPEDF-R(l-288) fold changes in the presence of different concentrations of H105A, Pl, P18, P19, and P24 peptides. The reaction was performed as in previous figures. The final concentration of rhuPEDF-R (1-288) was 60 nM. The plot represents an average of two independent experiments with reactions performed in triplicate for each concentration.

[0043] FIGS. 16A and 16B show PLAi (FIG. 16A) and PLA2(FIG. 16B) activity of rhuPEDF-R(l- 288) in the presence of P56 (SEQ ID NO: 61), P57 (SEQ ID NO: 62), P58 (SEQ ID NO: 63), or P3’ (SEQ ID NO: 64) at 250 pM. Each bar represents the average of three independent experiments, each of which were performed in triplicate for each peptide. The final concentration of rhuPEDF-R[l-288] was 60 nM and the reaction volume was 100 pl.

[0044] FIGS. 17A-17D show effects of peptides P24 (SEQ ID NO: 26) and P31 (SEQ ID NO: 33) in rdlO mice. The experimental timeline was the same as shown in FIG. 6A. FIG. 17A is a series of fluorescence fundoscopy images showing PS externalization of retinas from rd 10 mice treated with eyedrops containing the indicated peptide at 1 mg / ml. The contralateral eye received vehicle control (H2O). FIG. 17B is a series of graphs showing quantification of fluorescence PS externalization. Mean intensity of fluorescence of eyes treated with H2O was subtracted from the fluorescence of eyes treated with peptide and plotted. N=3 and each data point represents average ± SD of fluorescence relative to water by unpaired t-test. *** p<0.0001. FIG. 17C shows images of retina sections of rdlO mice at P21 stained with hematoxylin and eosin. OS, outer segment; ONL, outer nuclear layer; INL, inner nuclear layer. FIG. 17D is a spider plot of the thickness of the ONL as a function of distance from the optic nerve (ON). For all histology shown, three retinas per group and two sections per retina were evaluated and each data point corresponds to the average ± SD of ONL thickness per location by unpaired t-test. * p< 0.05, **p < 0.001, ***p < 0.0001, ****p < 0.00001.

[0045] FIGS. 18A-18D show effect of peptides Pl (SEQ ID NO: 3), P18 (SEQ ID NO: 20), P19 (SEQ ID NO: 21), and P5 (SEQ ID NO: 7) on apoptosis markers in rdlO mouse retinas. The experimental timeline was the same as shown in FIG. 6 A. FIG. 18A shows fluorescent micrographs of retinal sections from rdlO mice treated with eye drops of each peptides Pl, P18, P19, and P5 at 1 mg / ml per mouse at P21 days of age. Each contralateral eye was treated with vehicle (H2O). Immunostaining was performed with antibodies against BAX and DAPI. FIG. 18B shows fluorescent micrographs of retinas from rdlO mice treated with eye drops of each peptide Pl, P18, P19, or P5 at 1 mg / ml per mouse. Each contralateral eye was treated with vehicle (H2O). Immunostaining was performed with antibodies against BCL2 and DAPI. FIG. 18C is a histogram showing quantification of immunofluorescence intensity for BAX in retinal sections. FIG. 18D is a histogram showing quantification of immunofluorescence for BCL2 in retinal sections. For FIGS. 18C and 18D, three retinas per group were analyzed, with two sections per retina. Each individual data point is the average of the intensity of immunostaining from three ROIs from one retina, with statistical significance determined by unpaired t-test (***p < 0.0001, ****p < 0.00001). Scale bar = 20 pm.

[0046] SEQUENCE LISTING

[0047] The nucleic and amino acid sequences listed in the accompanying sequence listing are shown using standard letter abbreviations for nucleotide bases, and single letter code for amino acids, as defined in 37 C.F.R. 1.822. Only one strand of each nucleic acid sequence is shown, but the complementary strand is understood as included by any reference to the displayed strand.

[0048] SEQ ID NO: 1 is a wild type PEDF 17mer peptide shown in Table 1.

[0049] SEQ ID NO: 2 is a PEDF H105A peptide shown in Table 1.

[0050] SEQ ID NOs: 3-54 are exemplary modified PEDF peptides shown in Table 1.

[0051] SEQ ID NO: 55 is an exemplary human PEDF nucleic acid sequence:

[0052] ATGCAGGCCCTGGTGCTACTCCTCTGCATTGGAGCCCTCCTCGGGCACAGCAGC TGCCAGAACCCTGCCAGCCCCCCGGAGGAGGGCTCCCCAGACCCCGACAGCAC AGGGGCGCTGGTGGAGGAGGAGGATCCTTTCTTCAAAGTCCCCGTGAACAAGC TGGCAGCGGCTGTCTCCAACTTCGGCTATGACCTGTACCGGGTGCGATCCAGCA TGAGCCCCACGACCAACGTGCTCCTGTCTCCTCTCAGTGTGGCCACGGCCCTCT CGGCCCTCTCGCTGGGAGCGGAGCAGCGAACAGAATCCATCATTCACCGGGCT CTCTACTATGACTTGATCAGCAGCCCAGACATCCATGGTACCTATAAGGAGCTC

[0053] CTTGACACGGTCACTGCCCCCCAGAAGAACCTCAAGAGTGCCTCCCGGATCGTC

[0054] TTTGAGAAGAAGCTGCGCATAAAATCCAGCTTTGTGGCACCTCTGGAAAAGTCA

[0055] TATGGGACCAGGCCCAGAGTCCTGACGGGCAACCCTCGCTTGGACCTGCAAGA

[0056] GATCAACAACTGGGTGCAGGCGCAGATGAAAGGGAAGCTCGCCAGGTCCACAA

[0057] AGGAAATTCCCGATGAGATCAGCATTCTCCTTCTCGGTGTGGCGCACTTCAAGG

[0058] GGCAGTGGGTAACAAAGTTTGACTCCAGAAAGACTTCCCTCGAGGATTTCTACT

[0059] TGGATGAAGAGAGGACCGTGAGGGTCCCCATGATGTCGGACCCTAAGGCTGTT

[0060] TTACGCTATGGCTTGGATTCAGATCTCAGCTGCAAGATTGCCCAGCTGCCCTTG

[0061] ACCGGAAGCATGAGTATCATCTTCTTCCTGCCCCTGAAAGTGACCCAGAATTTG

[0062] ACCTTGATAGAGGAGAGCCTCACCTCCGAGTTCATTCATGACATAGACCGAGAA

[0063] CTGAAGACCGTGCAGGCGGTCCTCACTGTCCCCAAGCTGAAGCTGAGTTACGAA

[0064] GGCGAAGTCACCAAGTCCCTGCAGGAGATGAAGCTGCAATCCTTGTTTGATTCA

[0065] CCAGACTTTAGCAAGATCACAGGCAAACCCATCAAGCTGACTCAGGTGGAACA

[0066] CCGGGCTGGCTTTGAGTGGAACGAGGATGGGGCGGGAACCACCCCCAGCCCAG

[0067] GGCTGCAGCCTGCCCACCTCACCTTCCCGCTGGACTATCACCTTAACCAGCCTTT

[0068] CATCTTCGTACTGAGGGACACAGACACAGGGGCCCTTCTCTTCATTGGCAAGAT

[0069] TCTGGACCCCAGGGGCCCCTAA

[0070] SEQ ID NO: 56 is the amino acid sequence of an exemplary human PEDF protein:

[0071] MQALVLLLCIGALLGHSSCQNPASPPEEGSPDPDSTGALVEEEDPFFKVPVNKLAAA

[0072] VSNFGYDLYRVRSSMSPTTNVLLSPLSVATALSALSLGAEQRTESIIHRALYYDLISS

[0073] PDIHGTYKELLDTVTAPQKNLKSASRIVFEKKLRIKSSFVAPLEKSYGTRPRVLTGNP

[0074] RLDLQEINNWVQAQMKGKLARSTKEIPDEISILLLGVAHFKGQWVTKFDSRKTSLY

[0075] LDEERTVRVPMMSDPKAVLRYGLDSDLSCKIAQLPLTGSMSIIFFLPLKVTQNLTLIE

[0076] ESLTSEFIHDIDRELKTVQAVLTVPKLKLSYEGEVTKSLQEMKLQSLFDSPDFSKITG

[0077] KPIKLTQVEHRAGFEWNEDGAGTTPSPGLQPAHLTFPLDYHLNQPFIFVLRDTDTGA

[0078] LLFIGKILDPRGP

[0079] SEQ ID NO: 57 is the amino acid sequence of PEDF R99A peptide:

[0080] QATESIIHRALYYDLIS

[0081] SEQ ID NO: 58 is a nucleic acid sequence encoding PEDF-R(l-288):

[0082] ATGTTTCCCCGCGAGAAGACGTGGAACATCTCGTTCGCGGGCTGCGGCTTCCTC

[0083] GGCGTCTACTACGTCGGCGTGGCCTCCTGCCTCCGCGAGCACGCGCCCTTCCTG

[0084] GTGGCCAACGCCACGCACATCTACGGCGCCTCGGCCGGGGCGCTCACGGCCAC

[0085] GGCGCTGGTCACCGGGGTCTGCCTGGGTGAGGCTGGTGCCAAGTTCATTGAGGT ATCTAAAGAGGCCCGGAAGCGGTTCCTGGGCCCCCTGCACCCCTCCTTCAACCT GGTAAAGATCATCCGCAGTTTCCTGCTGAAGGTCCTGCCTGCTGATAGCCATGA GCATGCCAGTGGGCGCCTGGGCATCTCCCTGACCCGCGTGTCAGACGGCGAGA ATGTCATTATATCCCACTTCAACTCCAAGGACGAGCTCATCCAGGCCAATGTCT

[0086] GCAGCGGTTTCATCCCCGTGTACTGTGGGCTCATCCCTCCCTCCCTCCAGGGGGT

[0087] GCGCTACGTGGATGGTGGCATTTCAGACAACCTGCCACTCTATGAGCTTAAGAA

[0088] CACCATCACAGTGTCCCCCTTCTCGGGCGAGAGTGACATCTGTCCGCAGGACAG

[0089] CTCCACCAACATCCACGAGCTGCGGGTCACCAACACCAGCATCCAGTTCAACCT GCGCAACCTCTACCGCCTCTCCAAGGCCCTCTTCCCGCCGGAGCCCCTGGTGCT GCGAGAGATGTGCAAGCAGGGATACCGGGATGGCCTGCGCTTTCTGCAGCGGA

[0090] ACGGCCTCCTGAACCGGCCCAACCCCTTGCTGGCGTTGCCCCCCGCCCGCCCCC ACGGCCCAGAGGACAAGGACCAGGCAGTGGAGAGCGCCCAAG CGGAGGATTACTCG

[0091] SEQ ID NO: 59 is the amino acid sequence of PEDF-R(l-288):

[0092] MFPREKTWNISFAGCGFLGVYYVGVASCLREHAPFLVANATHIYGASAGALTATA

[0093] LVTGVCLGEAGAKFIEVSKEARKRFLGPLHPSFNLVKIIRSFLLKVLPADSHEHASGR

[0094] LGISLTRVSDGENVIISHFNSKDELIQANVCSGFIPVYCGLIPPSLQGVRYVDGGISDN

[0095] LPLYELKNTITVSPFSGESDICPQDSSTNIHELRVTNTSIQFNLRNLYRLSKALFPPEPL

[0096] VLREMCKQGYRDGLRFLQRNGLLNRPNPLLALPPARPHGPEDKDQAVESAQAEDY S

[0097] SEQ ID NO: 60 is the amino acid sequence of a 6X-SUMO-PEDF-R(l-288)-2X Strep construct (amino acids 1-6: 6X His tag; amino acids 7-109: small ubiquitin-related modifier (SUMO); amino acids 110-397: PEDF-R(l-288); amino acids 398-413: 2X strep tag):

[0098] HHHHHHMSEEKPKEGVKTENDHINLKVAGQDGSVVQFEIKRHTPLSKLMKAYCER QGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGGVPESSLAGHSFMFP REKTWNISFAGCGFLGVYYVGVASCLREHAPFLVANATHIYGASAGALTATALVT

[0099] GVCLGEAGAKFIEVSKEARKRFLGPLHPSFNLVKIIRSFLLKVLPADSHEHASGRLGI

[0100] SLTRVSDGENVIISHFNSKDELIQANVCSGFIPVYCGLIPPSLQGVRYVDGGISDNLPL YELKNTITVSPFSGESDICPQDSSTNIHELRVTNTSIQFNLRNLYRLSKALFPPEPLVL REMCKQGYRDGLRFLQRNGLLNRPNPLLALPPARPHGPEDKDQAVESAQAEDYSW SHPQFEKWSHPQFEK

[0101] SEQ ID NOs: 61-64 are additional exemplary modified PEDF peptides shown in Table 1. DETAILED DESCRIPTION

[0102] Described herein are modified PEDF peptides that are effective in treating or inhibiting retinal degeneration in two mouse models. Peptide drug development has made great progress recently thanks to new production, modification, and analytic technologies. Solutions of chemically synthesized bioactive peptides have unique advantages over mixed formulations. They are free of inactive ingredients that may cause secondary effects. Structural biology and recombinant biologies aid in peptide design to modify amino acids to enhance target affinity, specificity, regulate bioactivity, as well as in improved solubility, and stability of the peptide drugs. Peptides can diffuse better than biologies (proteins, antibodies) with better penetrability. Compared with biologies, therapeutic peptides have demonstrably less immunogenicity and lower production costs.

[0103] I. Terms

[0104] Unless otherwise noted, technical terms are used according to conventional usage. Definitions of many common terms in molecular biology may be found in Krebs et al. (eds.), Lewin 's genes XII, published by Jones & Bartlett Learning, 2017. As used herein, the singular forms “a,” “an,” and “the,” refer to both the singular as well as plural, unless the context clearly indicates otherwise. For example, the term “a peptide” includes singular or plural peptides and can be considered equivalent to the phrase “at least one peptide.” As used herein, the term “comprises” means “includes.” It is further to be understood that any and all base sizes or amino acid sizes, and all molecular weight or molecular mass values, given for nucleic acids or polypeptides are approximate, and are provided for descriptive purposes, unless otherwise indicated.

[0105] When chemical structures are depicted or described, unless explicitly stated otherwise, all carbons are assumed to include hydrogen so that each carbon conforms to a valence of four. For example, in the structure on the left-hand side of the schematic below there are nine hydrogen atoms implied. The nine hydrogen atoms are depicted in the right-hand structure.

[0106] Sometimes a particular atom in a structure is described in textual formula as having a hydrogen or hydrogen atoms, for example -CH2CH2-. It will be understood by a person of ordinary skill in the art that the aforementioned descriptive techniques are common in the chemical arts to provide brevity and simplicity to description of organic structures. Although many methods and materials similar or equivalent to those described herein can be used, particular suitable methods and materials are described herein. In case of conflict, the present specification, including explanations of terms, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. To facilitate review of the various aspects, the following explanations of terms are provided:

[0107] Aliphatic: A substantially hydrocarbon-based group or moiety. An aliphatic group or moiety can be acyclic, including alkyl, alkenyl, or alkynyl groups, cyclic versions thereof, such as cycloaliphatic groups or moieties including cycloalkyl, cycloalkenyl or cycloalkynyl, and further including straight- and branched-chain arrangements, and all stereo and position isomers as well. Unless expressly stated otherwise, an aliphatic group contains from one to twenty-five carbon atoms (C1-25); for example, from one to fifteen (C1-15), from one to ten (C1-10), from one to six (Cn e), or from one to four carbon atoms (Ci^i) for an acyclic aliphatic group or moiety, or from three to fifteen (C3-15) from three to ten (C3-10), from three to six (C3-6), or from three to four (C3-4) carbon atoms for a cycloaliphatic group or moiety. An aliphatic group may be substituted or unsubstituted, unless expressly referred to as an “unsubstituted aliphatic” or a “substituted aliphatic.” An aliphatic group can be substituted with one or more substituents (up to two substituents for each methylene carbon in an aliphatic chain, or up to one substituent for each carbon of a -C=C- double bond in an aliphatic chain, or up to one substituent for a carbon of a terminal methine group).

[0108] Alkyl: A hydrocarbon group having a saturated carbon chain. The chain may be cyclic (e.g. cycloalkyl), branched or unbranched. Examples, without limitation, of alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl and decyl. Unless expressly stated otherwise, an aliphatic group contains from 1-12 carbon atoms such as from one to ten (C1-10), from one to six (Cue), or from one to four carbon atoms (CM) for an acyclic alkyl group or moiety, or from three to twelve (C3-12) from three to ten (C3-10), from three to six (C3-6), or from three to four (C3-4) carbon atoms for a cycloalkyl group or moiety. The terms alkenyl and alkynyl refer to hydrocarbon groups having carbon chains containing one or more double or triple bonds, respectively, and may have from 2-12 carbon atoms.

[0109] Aromatic: A cyclic, conjugated group or moiety of, unless specified otherwise, from 5 to 15 ring atoms having a single ring (e.g., phenyl, pyridinyl, or pyrazolyl) or multiple condensed rings in which at least one ring is aromatic (e.g., naphthyl, indolyl, or pyrazolopyridinyl), that is at least one ring, and optionally multiple condensed rings, have a continuous, delocalized 71-electron system. Typically, the number of out of plane 71-electrons corresponds to the Hiickel rule (4n + 2). The point of attachment to the parent structure typically is through an aromatic portion of the condensed ring system. For example, . However, in certain examples, context or express disclosure may indicate that the point of attachment is through a non-aromatic portion of the condensed ring system. For example, ■ . An aromatic group or moiety may comprise only carbon atoms in the ring, such as in an aryl group or moiety, or it may comprise one or more ring carbon atoms and one or more ring heteroatoms comprising a lone pair of electrons (e.g. S, O, N, P, or Si), such as in a heteroaryl group or moiety. Unless otherwise stated, an aromatic group may be substituted or unsubstituted.

[0110] A wave line “ «A / W* ” on a group or moiety indicates a point of attachment from the group or moiety to the rest of the compound.

[0111] Aryl: A monovalent aromatic carbocyclic group of, unless specified otherwise, from 6 to 15 carbon atoms having a single ring (e.g., phenyl) or multiple condensed rings in which at least one ring is aromatic (e.g. , indole, benzodioxole, and the like), provided that the point of attachment is through an atom of an aromatic portion of the aryl group and the aromatic portion at the point of attachment contains only carbons in the aromatic ring. If any aromatic ring portion contains a heteroatom, the group is a heteroaryl and not an aryl. Aryl groups are monocyclic, bicyclic, tricyclic or tetracyclic.

[0112] Heteroaryl: A monovalent monocyclic or bicyclic aromatic moiety of 5 to 12 ring atoms with each ring containing one, two, three or four ring heteroatoms independently selected from N, O, or S, the remaining ring atoms being C. The aromatic moiety is optionally fused to a phenyl or an optionally substituted heterocycle ring. In specific examples, the term heteroaryl includes, but is not limited to pyridyl, pyrrolyl, thiophenyl, pyrazolyl, thiazolyl, imidazolyl, pyrimidinyl, thiadiazolyl, indolyl, carbazolyl, azaindolyl, benzofuranyl, benzimidazolyl, benzthiazolyl, quinoxalinyl, benzotriazolyl, benzisoxazolyl, purinyl, quinolinyl, isoquinolinyl, benzopyranyl, and derivatives thereof.

[0113] “Heterocycle” refers to both aromatic and non-aromatic ring systems, and more specifically refer to a stable three- to fifteen-membered ring moiety comprising at least one carbon atom, and typically plural carbon atoms, and at least one, such as from one to five, heteroatoms. The heteroatom(s) independently may be nitrogen, phosphorus, oxygen, silicon or sulfur atom(s). The heterocycle moiety may be a monocyclic moiety, or may comprise multiple rings, such as in a bicyclic or tricyclic ring system, provided that at least one of the rings contains a heteroatom. Such a multiple ring moiety can include fused or bridged ring systems as well as spirocyclic systems; and any nitrogen, phosphorus, carbon, silicon or sulfur atoms in the heterocycle moiety can be optionally oxidized to various oxidation states. For convenience, nitrogens, particularly, but not exclusively, those defined as annular aromatic nitrogens, are meant to include their corresponding N-oxide form, although not explicitly defined as such in a particular example. Thus, for a compound having, for example, a pyridinyl ring, the corresponding pyridinyl-N-oxide is included as another compound, unless expressly excluded or excluded by context. In addition, annular nitrogen atoms can be optionally quaternized. Heterocycle includes heteroaryl moieties, and heterocycloaliphatic moieties, which are heterocycle rings that are partially or fully saturated. Examples of heterocycle groups include, but are not limited to, azetidinyl, oxetanyl, acridinyl, benzodioxolyl, benzodioxanyl, benzofuranyl, carbazoyl, cinnolinyl, dioxolanyl, indolizinyl, naphthyridinyl, perhydroazepinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrazoyl, tetrahydroisoquinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2- oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, dihydropyridinyl, tetrahydropyridinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl, octahydroindolyl, octahydroisoindolyl, quinolyl, isoquinolyl, decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, diazabicycloheptane, diazapane, diazepine, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothieliyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, dioxaphospholanyl, and oxadiazolyl.

[0114] Macular degeneration: A condition in which the cells of the macula (the central part of the retina) degenerate, resulting in loss of central visual acuity. The most common form of macular degeneration is age-related macular degeneration (AMD). AMD is the most common cause of irreversible loss of central vision and legal blindness in the elderly. It causes progressive damage to the macula, resulting in gradual loss of central vision. There are two forms, atrophic and neovascular macular degeneration. In atrophic degeneration (dry form), the tissues of the macula thin as photoreceptor cells disappear. There is currently no treatment for atrophic degeneration, though dietary supplements may help slow progression. In neovascular macular degeneration (wet form), abnormal blood vessels develop under the macula. These vessels may leak fluid and blood under the retina and eventually a mound of scar tissue develops under the retina. Central vision becomes washed out and loses detail, and straight lines may appear wavy. For neovascular macular degeneration there are some treatments available, including the use of medication injected directly into the eye e.g., anti-VEGF therapy), laser therapy in combination with a targeting drug (e.g., photodynamic therapy) and brachytherapy. However, repeated treatments can cause complications leading to loss of vision.

[0115] Stargardt-like macular degeneration (STGD3) is an early onset, autosomal dominant macular degeneration. STGD3 is characterized by a progressive pathology, the loss of central vision, atrophy of the retinal pigment epithelium, and accumulation of lipofuscin, clinical features that are also characteristic of AMD.

[0116] Maculopathy: Any pathological condition of the macula. Maculopathies include AMD and STGD3, as well as vitelliform macular dystrophy (Best disease), Malattia Leventinese (Doyne’s honeycomb retinal dystrophy), diabetic maculopathy, occult macular dystrophy, and cellophane maculopathy.

[0117] Pigment epithelium-derived factor (PEDF): Also referred to as SERPINF1 (serpin family F member 1). A member of the serpin family that does not exhibit demonstrable serpin inhibitory activity against proteases. PEDF is a secreted protein with anti- angiogenic and neurotrophic properties. PEDF binds to PEDF-R (also known as PNPLA2), which exhibits phospholipase A (PLA) activity (such as PLAi and / or PLA2 activity) upon PEDF binding. In some examples, PEDF binds to PEDF-R(l-288) (for example, SEQ ID NO: 59), and stimulates PLAi and / or PLA2 activity.

[0118] PEDF nucleic acid and protein sequences are publicly available. Human PEDF nucleic acid sequences include GenBank Accession Nos. NM_002615.7, NM_001329904.2, and NM_001329903.2 (all of which are incorporated by reference as present in GenBank on November 27, 2023), and SEQ ID NO: 55. Human PEDF amino acid sequences include GenBank Accession Nos. NP_002606.3, NP_001316833.1, and NP_001316832.1 (all of which are incorporated by reference as present in GenBank on November 27, 2023), and SEQ ID NO: 56.

[0119] Retinal Degeneration: Deterioration of the retina, including progressive death of the photoreceptor cells of the retina or associated structures (such as retinal pigment epithelium). Retinal degeneration includes diseases or conditions such as retinitis pigmentosa, cone-rod dystrophy, macular degeneration (such as age-related macular degeneration and Stargardt- like macular degeneration), Leber congenital amaurosis (LCA), and maculopathies.

[0120] Retinitis pigmentosa (RP): An inherited, degenerative eye disease that causes severe vision impairment due to the progressive degeneration of the rod photoreceptor cells in the retina. This form of retinal dystrophy manifests initial symptoms independent of age. The initial retinal degenerative symptoms of RP are characterized by decreased night vision (nyctalopia) and the loss of the mid-peripheral visual field. The rod photoreceptor cells, which are responsible for low-light vision and are orientated in the retinal periphery, are the retinal processes affected first during non- syndromic forms of this disease. Visual decline progresses relatively quickly to the far peripheral field, eventually extending into the central visual field as tunnel vision increases. Visual acuity and color vision can become compromised due to accompanying abnormalities in the cone photoreceptor cells, which are responsible for color vision, visual acuity, and sight in the central visual field. The progression of disease symptoms occurs in a symmetrical manner, with both the left and right eyes experiencing symptoms at a similar rate. There are multiple genes that, when mutated, can cause the retinitis pigmentosa phenotype. Inheritance patterns of RP have been identified as autosomal dominant, autosomal recessive, X-linked, and maternally (mitochondrially) acquired, and are dependent on the specific RP gene mutations present in the parental generation.

[0121] Subject: A living multi-cellular vertebrate organism, a category that includes human, laboratory, and veterinary subjects, including human and non-human mammals.

[0122] Therapeutically effective amount: An amount of a compound sufficient to treat a specified disorder or disease, or to ameliorate or eradicate one or more of its symptoms and / or to prevent the occurrence of the disease or disorder, such as retinal degeneration. The amount of a compound which constitutes a “therapeutically effective amount” will vary depending on the compound, the route of administration, the disease state and its severity, the age of the subject to be treated, and the like. The therapeutically effective amount can be determined by a person of ordinary skill in the art, for example, through various in vitro, in vivo, or ex vivo assays.

[0123] Treating, Treatment, and Therapy: Any success or indicia of success in the attenuation or amelioration of an injury, pathology, or condition, including any objective or subjective parameter such as abatement, remission, diminishing of symptoms or making the condition more tolerable to the subject, slowing in the rate of degeneration or decline, making the final point of degeneration less debilitating, improving a subject’s physical or mental well-being, or improving vision. The treatment may be assessed by objective or subjective parameters; including the results of a physical examination, neurological examination, or psychiatric evaluations. The term “ameliorating,” with reference to a disease or pathological condition, refers to any observable beneficial effect of the treatment. The beneficial effect can be evidenced, for example, by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in severity of some or all clinical symptoms of the disease, a slower progression of the disease, an improvement in the overall health or well-being of the subject, or by other parameters known in the art that are specific to the particular disease, such as improved vision. A “prophylactic” treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing pathology. II. Modified PEDF peptides

[0124] Disclosed herein are modified peptides that include one or more modifications compared to the PEDF 17mer peptide SEQ ID NO: 1. In some aspects the modified peptide includes SEQ ID NO: 1 with one or more modifications, wherein the one or more modifications are selected from: a) an N-terminal modification; b) one or more amino acid substitutions; c) deletion of 1-4 amino acids from the N-terminus and / or C-terminus of the peptide; d) replacement of one or more amino acids with a non-alpha-amino acid linker; e) a C-terminal modification; and f) any combination of two or more thereof, and wherein the peptide includes 8-19 amino acids. In some examples, the peptide includes 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 amino acids (such as 8-13, 12-18 or 13-17 amino acids).

[0125] In some examples, the modified peptide includes one or more of the modifications listed above, such as 1, 2, 3, 4, 5, or more modifications. In some examples, the modified peptide includes at least one of each of a), b), and c); at least one each of a), b), and d); at least one of each of a), b), and e); at least one of each of a), d), and e); at least one of each of a), c), and e); at least one each of a), c), and d); at least one of each of a), b), c), and d); at least one of each of a), b), d), and e); or at least one each of a), b), c), and e). In other examples, the modified peptide includes at least one modification of each of a)-e).

[0126] In some examples, the modified peptide includes an N-terminal modification. In some examples, each N-terminal modification independently comprises a moiety selected from -C Oi-R1, -QOfNH-R1, -C(O)-Ar', -C(O)NH-Ar’, -C(O)-(CH2)xAr1or - OlNH- Cf fxAr1, wherein:

[0127] R1is optionally substituted C1-12 alkyl;

[0128] Ar1is optionally substituted Ce-io aryl or optionally substituted 5- to 12-membered heterocycle comprising 1, 2, 3, or 4 heteroatoms selected from N, S, and / or O. In some examples, R1is a nitrogen-containing heterocycle; and x is from 1 to 12.

[0129] In some examples, R1is optionally substituted C1-6 alkyl.

[0130] In some examples, R1is unsubstituted, but in other examples, R1is substituted, such as by NH2or OH.

[0131] In some examples, Ar1is Ce-io aryl, such as Ce-io aryl, and may be phenyl or naphthyl.

[0132] In some examples, Ar1is 5- to 10-membered heterocycle, such as a 5- to 10-membered heteroaryl. In some examples, Ar1is a 5- or 6-membered heterocycle, such as a 5- to 6-membered heteroaryl. In some examples, Ar1is pyridinyl, pyrimidinyl, pyrazolyl, imidazolyl, pyrrolyl, triazolyl, tetrazolyl, indolyl, quinolinyl, thiazolyl, oxazolyl, isoxazolyl, furanyl, benzofuranyl, benzoimidazolyl, pyridylimidazolyl.

[0133] In some examples, Ar1is unsubstituted, but in other examples, Ar1is substituted, such as with Ci-ealkyl, fluoro, chloro, bromo, iodo, -OR’, -NHR’, or -C(O)NHR’ where R’ is H or alkyl, such as H or Ci-6alkyl.

[0134] In some examples, the N-terminal modification is 0 (derived from 6- aminohexanoic acid (AHA)), O (derived from 5 -aminopentanoic acid (APA)),

[0135] O (derived from 4- aminobutanoic acid (ABA)), or O (derived from beta-alanine (b-Ala)).

[0136] In some examples, the modified peptide includes one or more amino acid substitutions compared to the amino acid sequence of SEQ ID NO: 1. The one or more amino acid substitutions may include substitution of an amino acid present in SEQ ID NO: 1 with a different L-amino acid, a D-amino acid, a modified amino acid, or a non-naturally occurring (or unnatural) amino acid. In some examples, the modified peptide is not SEQ ID NO: 2. In other examples, the modified peptide is not SEQ ID NO: 57.

[0137] In some examples, the one or more amino acid substitutions are at one or more positions corresponding to amino acid positions 5, 6, and / or 8 of SEQ ID NO: 1. In some examples, the one or more amino acid substitutions include substitution of a threonine or aspartic acid for serine at a position corresponding to amino acid position 5 of SEQ ID NO: 1. In other examples, the one or more amino acid substitutions include substitution of diaminopropionic acid (DAP) or CF3- threonine for serine at a position corresponding to amino acid position 5 of SEQ ID NO: 1.

[0138] In other examples, the one or more amino acid substitutions include substitution of norleucine, leucine, valine, or N-methylated isoleucine for isoleucine at a position corresponding to amino acid position 6 of SEQ ID NO: 1.

[0139] In additional examples, the one or more amino acid substitutions include substitution of alanine for histidine at a position corresponding to amino acid position 8 of SEQ ID NO: 1. In particular examples, the modified peptide is not SEQ ID NO: 1 with substitution of alanine at the position corresponding to amino acid position 8 of SEQ ID NO: 1 and no other modifications, e.g. , the modified peptide is not SEQ ID NO: 2. In further examples, the peptide includes replacement of one or more (such as 1, 2, 3, 4, or more) amino acids with a non-alpha amino acid linker (also referred to as the “linker”). In some examples, the linker has a formula -NR’-R2-C(O)-, wherein:

[0140] -R2- is -(CR32)p-, -(CH2CH2O)yCH2-, -(CH2)ZI-Y-(CH2)Z2-, -(CH2)zi-Ar2-(CH2)Z2->-(CH2)ZI- Ar2-Ar3-(CH2)z2-, or -(CH2)zi-Ar2-X-Ar3-(CH2)z2-; p is from 1 to 12, wherein if p is 1, at least one R3is -CH(OH)(CF3); each R3independently is H, Ci-ealkyl, OH, -CH(OH)(CF3), NH2or -CH2NH2; y is from 1 to 6, such as from 1 to 4, and in certain examples, y is 2 or 3 or 4, and may be 2; each of zl and z2 independently is from 0 to 6, such as from 0 to 4, or from 0 to 2;

[0141] X is O, NR’, or -Ci-ealkyl-, such as O or NH;

[0142] Y is O or NR’ ; each R’ independently is H or Ci-ealkyl; and each of Ar2and Ar3independently is optionally substituted Ce-io aryl, or optionally substituted 5- to 12-membered heterocycle comprising 1, 2, 3, or 4 heteroatoms selected from N, S, and / or O. In some examples, Ar2and / or Ar3is a nitrogen-containing heterocycle.

[0143] In some examples, -R2- is -(CR32)P-. In some examples, p is from 2 to 12, such as from 2 to 8, or from 2 to 6, and may be 4, 5 or 6. In some examples, each R3is H, but in other examples, one or more R3is other than H, such as 1 or 2 R3, or one R3is other than H, and may be Ci-ealkyl, OH, NH2or -CH2NH2. In certain examples, 1, 2, or 3 R3independently are selected from OH, or NH2and the remaining R3are H.

[0144] In some examples, Ar2is Ce-io aryl and may be phenyl or naphthyl.

[0145] In some examples, Ar2is 5- to 12-membered heterocycle, such as a 5- to 10-membered heterocycle, or a 5- to 6-membered heterocycle. In some examples, Ar2is a 5- to 12-membered heteroaryl, such as a 5- to 10-membered heteroaryl, or a 5- to 6-membered heteroaryl. In certain examples, Ar2is pyridinyl.

[0146] In some examples, Ar3is Ce-ioaryl and may be phenyl. In other examples, Ar3is 5- to 12- membered heterocycle, such as a 5- to 10-membered heterocycle, or a 5- to 6-membered heterocycle. In some examples, Ar3is a 5- to 12-membered heteroaryl, such as a 5- to 10- membered heteroaryl, or a 5- to 6-membered heteroaryl. In certain examples, Ar3is pyridinyl.

[0147] In some examples, Ar2and Ar3are both phenyl.

[0148] In some examples, zl and z2 are the same but in other examples, zl and z2 are different. In some examples, zl and z2 are both 0. In other examples, one of zl and z2 is 0 and the other is from 1 to 4, such as from 1 to 2. In certain examples, one of zl and z2 is 0 and the other of zl and z2 is 1. And in a particular example, zl is 1 and z2 is 0. In some examples, Ar2is unsubstituted, but in other examples Ar2is substituted, such as by Ci -6 alkyl, fluoro, chloro, bromo, iodo, -OR’, -NHR’, or -C(O)NHR’ where R’ is H or alkyl, such as H or Ci-6alkyl.

[0149] In some examples, Ar3is unsubstituted, but in other examples Ar3is substituted, such as by Ci-6 alkyl, fluoro, chloro, bromo, iodo, -OR’, -NHR’, or -C(O)NHR’ where R’ is H or alkyl, such as H or Ci-6alkyl. ln some examples, the linker is derived from 6-aminohexanoic acid, 6-aminopentanoic acid), 8-amino-3,6,dioxaoctanoic acid (AOC), diaminopropionic acid (DAP), 4’-amino-[l,l’- biphenyl]-4-carboxylate (AB PC), 4-(4-(aminomethyl)phenoxy)benzoic acid, 8-aminooctanoic acid (AOA), or P-alanine.

[0150] In some examples, the linker is (AHA),

[0151] In some examples, the peptide includes replacement of at least amino acid 8, 9, or 10 of SEQ ID NO: 1 with the linker. In particular examples, the peptide further includes replacement of one amino acid with a linker and deletion of 1-3 amino acids following the position of substitution with the non-alpha-amino acid linker (e.g., replacement of 2-4 amino acids with the linker).

[0152] In some examples, the peptide includes a C-terminal modification. In some examples, the C-terminal modification is -NH-R4where R4is Ci-ealkyl, optionally substituted with OH, or G,.

[0153] H H waryl. In some examples, the C-terminal modification is selected from , or In other examples, the C-terminal modification is C-terminal amidation.

[0154] In some examples, the modified peptide includes an N-terminal modification, one or more amino acid substitutions, and C-terminal amidation. In other examples, the modified peptide includes N-terminal modification, deletion of 1-4 amino acids from the N-terminus, and C-terminal amidation. In other examples, the modified peptide includes N-terminal modification, one or more amino acid substitutions, deletion of 1-4 amino acids from the N-terminus, and C-terminal amidation. In additional examples, the peptide includes N-terminal modification, deletion of 1-4 amino acids from the C-terminus, and C-terminal amidation. In additional examples, the peptide includes N-terminal modification, substitution of one or more amino acids, deletion of 1-4 amino acids from the C-terminus, and C-terminal amidation. In other examples, the modified peptide includes N-terminal modification, replacement of one or more amino acids with a non- alpha- amino acid linker, and C-terminal amidation. In other examples, the modified peptide includes N-terminal modification, one or more amino acid substitutions, replacement of one or more amino acids with a non-alpha- amino acid linker, and C-terminal amidation.

[0155] In some examples, a disclosed PEDF peptide increases phospholipase Ai activity of PEDF- R(l-288), phospholipase A2 activity of PEDF-R( 1-288), or both, compared to a control. In some examples the PEDF peptide increases phospholipase A2 activity of PEDF-R(l-288) by at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5- fold, at least about 4-fold, at least about 4.5-fold, a at least bout 5-fold, at least about 5.5-fold, at least about 6-fold, at least about 6.5-fold, at least about 7-fold, at least about 7.5 fold, or at least about 8-fold compared to a control. In other examples, the PEDF peptide increases phospholipase A2 activity of PEDF-R(l-288) by at least about 10-fold or more compared to a control In some examples, the PEDF peptide increases phospholipase Ai activity of PEDF-R( 1-288) by at least about 5-fold, at least about 7.5-fold, at least about 10-fold, at least about 15-fold, at least about 20- fold, at least about 25-fold, at least about 30-fold, at least about 40-fold, or more, compared to a control. In some examples, the control is a sample treated in the same way but without inclusion of a PEDF peptide. In other examples, the control is a sample treated in the same way, but with H105A peptide. Methods of measuring phospholipase Ai and phospholipase A2 activity are known to those of ordinary skill in the art. Exemplary non-limiting methods are described in Example 3.

[0156] In some non-limiting examples, a peptide that increases phospholipase Ai activity of PEDF- R(l-288), phospholipase A2 activity of PEDF-R(l-288), or both is a modified PEDF peptide including one of more of an N-terminal modification comprising AHA, P-alanine, or a derivative thereof; a deletion of 1-4 amino acids from the N-terminal of SEQ ID NO: 1; an amino acid substitution of threonine at a position corresponding to amino acid 5 of SEQ ID NO: 1; or a nonalpha amino acid linker comprising AOC or a derivative thereof.

[0157] In specific examples, the modified PEDF peptides of the disclosure include those provided in Table 1. In some particular examples, the modified PEDF peptide is Pl (SEQ ID NO: 3), P24 (SEQ ID NO: 26), Pl 8 (SEQ ID NO: 20), P19 (SEQ ID NO: 21), PIO (SEQ ID NO: 12), P17 (SEQ ID NO: 19), or P28 (SEQ ID NO: 30). Table 1. Exemplary modified PEDF peptides

[0158] AHA: 6- aminohexanoic acid

[0159] APA: 5 -aminopentanoic acid

[0160] AOC: 8-amino-3,6-dioxaoctanoic acid

[0161] P-Ala = 3 -Aminopropanoic acid ABA = 4- Aminobutanoic acid

[0162] 4’ AB PC : 4’ -amino- [ 1,1’ -biphenyl] -4-carboxylate

[0163] 4’AMPC: 4-(4-(aminomethyl)phenoxy)benzoic acid

[0164] Abn: benzylamine

[0165] Nle: norleucine

[0166] Ac: Acetyl

[0167] AOA: 8-aminooctanoic acid

[0168] III. Methods of Treatment

[0169] Also provided are methods of treating a retinal disease or disorder, such as retinal degeneration or macular degeneration, with compositions including the modified PEDF peptides provided herein. In some examples, the methods include administering a composition including one or more modified PEDF peptides to one or both eyes of a subject with a retinal disease or disorder. In some examples, the subject has an inherited retinal degeneration, including, but not limited to retinitis pigmentosa (RP), Leber congenital amaurosis, or cone -rod dystrophy. In other examples, the subject has macular degeneration, such as age-related macular degeneration (AMD, for example, wet AMD or dry AMD), Stargardt-like macular degeneration, vitelliform macular dystrophy (Best disease), bull’s eye maculopathy, or other maculopathy. In further examples, the subject has glaucoma.

[0170] Also provided are methods of treating or inhibiting cancer, including administering a composition including a therapeutically effective amount of one or more modified PEDF peptides to a subject with cancer. In some examples, the cancer is a primary tumor, a recurrence, a metastasis, or any combination thereof.

[0171] In some examples, the subject with cancer has a solid tumor, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, and other sarcomas, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, lymphoid malignancy, pancreatic cancer, breast cancer (including basal breast carcinoma, ductal carcinoma and lobular breast carcinoma), lung cancers, ovarian cancer, prostate cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, salivary gland carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, pheochromocytomas sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, Wilms' tumor, cervical cancer, testicular tumor, seminoma, bladder carcinoma, and CNS tumors (such as a glioma, astrocytoma, medulloblastoma, craniopharyrgioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma and retinoblastoma). In particular examples, the subject with cancer has ovarian cancer, liver cancer (for example, hepatocellular carcinoma), pancreatic cancer, or melanoma.

[0172] In some examples, the subject with cancer has a hematological malignancy. Exemplary hematological malignancies include leukemias, including acute leukemias (such as Hq23-positive acute leukemia, acute lymphocytic leukemia (ALL), T-cell ALL, acute myelocytic leukemia, acute myelogenous leukemia (AML), and myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia), chronic leukemias (such as chronic myelocytic (granulocytic) leukemia, chronic myelogenous leukemia (CML), and chronic lymphocytic leukemia (CLL)), lymphoblastic leukemia, polycythemia vera, lymphoma, diffuse large B cell lymphoma, Burkitt lymphoma, T cell lymphoma, follicular lymphoma, mantle cell lymphoma, Hodgkin disease, non-Hodgkin lymphoma, multiple myeloma, Waldenstrom macroglobulinemia, heavy chain disease, myelodysplastic syndrome (MDS), hairy cell leukemia, and myelodysplasia.

[0173] In further examples, the disclosed peptides can be used in methods of treating or inhibiting diseases or disorders that involve cell death in the central nervous system. Exemplary disease or disorders that include cell death in the CNS include amyotrophic lateral sclerosis (ALS), Huntington’s disease, Alzheimer’s disease, Parkinson’s disease, and human immunodeficiency virus (HlV)-induced encephalitis, as well as CNS trauma.

[0174] Pharmaceutical compositions that include one or more of the PEDF polypeptides disclosed herein (such as 1, 2, 3, 4, 5, or more PEDF polypeptides) can be formulated with an appropriate carrier, depending upon the particular mode of administration chosen. See, e.g., Remington: The Science and Practice of Pharmacy, Adejare (Ed.), Academic Press, London, United Kingdom, 23rdEdition (2021). For instance, formulations usually include fluids that are pharmaceutically and physiologically acceptable fluid vehicles such as water, physiological saline, other balanced salt solutions (e.g., Hank’s balanced salt solution), artificial tears, aqueous dextrose, glycerol or the like. In addition to biologically-neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, pH buffering agents, or the like, for example sodium acetate or sorbitan monolaurate. Excipients that can be included are, for instance, other proteins, such as human serum albumin or plasma preparations.

[0175] Pharmaceutical compositions disclosed herein include those formed from pharmaceutically acceptable salts and / or solvates of the disclosed compounds. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids. Particular disclosed compounds possess at least one basic group that can form acid-base salts with acids. Examples of basic groups include, but are not limited to, amino and imino groups.

[0176] Examples of inorganic acids that can form salts with such basic groups include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid. Basic groups also can form salts with organic carboxylic acids, sulfonic acids, sulfo acids or phospho acids or N-substituted sulfamic acid, for example acetic acid, formic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2- acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, and, in addition, with amino acids, for example with a-amino acids, and also with methanesulfonic acid, ethanesulfonic acid, 2- hydroxymethanesulfonic acid, ethane- 1 ,2-disulfonic acid, benzenedisulfonic acid, 4- methylbenzenesulfonic acid, naphthalene-2- sulfonic acid, 2- or 3 -phosphoglycerate, glucose-6- phosphate or jV-cyclohexylsulfamic acid (with formation of the cyclamates) or with other acidic organic compounds, such as ascorbic acid. In particular, suitable salts include those derived from alkali metals such as potassium and sodium, alkaline earth metals such as calcium and magnesium, among numerous other acids well known in the pharmaceutical art.

[0177] Certain compounds include at least one acidic group that can form an acid-base salt with an inorganic or organic base. Examples of salts formed from inorganic bases include salts of the presently disclosed compounds with alkali metals such as potassium and sodium, alkaline earth metals, including calcium and magnesium and the like. Similarly, salts of acidic compounds with an organic base, such as an amine (as used herein terms that refer to amines should be understood to include their conjugate acids unless the context clearly indicates that the free amine is intended) are contemplated, including salts formed with basic amino acids, aliphatic amines, heterocyclic amines, aromatic amines, pyridines, guanidines and amidines. Of the aliphatic amines, the acyclic aliphatic amines, and cyclic and acyclic di- and tri- alkyl amines are particularly suitable for use in the disclosed compounds. In addition, quaternary ammonium counterions also can be used.

[0178] In some examples, the composition is administered topically to one or both eyes of a subject with a retinal disease or disorder. Topical preparations may include eye drops, ointments, sprays, and the like. In some examples, the composition is an eyedrop and the amount of the composition administered is selected based on the subject being treated. In some examples, the amount of the composition administered is about 1 pl to about 10 pl (such as about 1-3 pl, about 2-4 pl, about 3-5 pl, about 4-6 pl, about 5-7 pl, about 6-8 pl, about 7-9 pl, or about 8-10 pl), for example, about 5 pl, for example, if the subject is a rodent. In other examples, the amount of the composition administered is about 25 pl to about 100 pl (such as about 25-50 pl, about 50-75 pl, or about 75- 100 pl), for example, about 75-80 pl, for example, if the subject is a human or a pig.

[0179] In some examples, the disclosed compositions include a therapeutically effective amount of the PEDF peptide when administered topically. In some examples, a therapeutically effective amount of the PEDF peptide is about 1-10 pg of peptide (for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 pg of peptide or about 1-2.5, 2-5, 4-7.5, or 6-10 pg of peptide) when administered topically. In one example, a therapeutically effective amount of the PEDF peptide is about 5 pg, administered topically. In some examples, the composition includes about 0.1 mg / ml to about 2 mg / ml of the peptide (for example, about 0.1 mg / ml to about 0.5 mg / ml, about 0.25 mg / ml to about 1 mg / ml, about 0.75 mg / ml to about 1.5 mg / ml, or about 1.25 mg / ml to about 2 mg / ml peptide). In one example, the composition includes about 1 mg / ml of PEDF peptide. In other examples, the composition includes about 0.05 mg / ml to about 0.5 mg / ml of the peptide (for example, about 0.05 mg / ml to about 0.1 mg / ml, about 0.075 mg / ml to about 0.2 mg / ml, or about 0.1 mg / ml to about 0.5 mg / ml peptide). In one example, the composition includes about 0.067 mg / ml of PEDF peptide.

[0180] In other examples, the composition is administered by injection to one or both eyes of a subject with a retinal disease or disorder. In some examples, the composition is administered by injection in the vitreous (intravitreal injection), the subretinal space (subretinal injection), the suprachoroidal space, or the subconjunctiva. In some examples, the composition is administered by intravitreal injection and the volume injected is about 0.5-2 pl (such as about 0.5-1 pl or about 1-2 pl) if the subject is a rodent or the volume injected is about 25-200 pl (for example, about 25-50 pl, about 50-100 pl, about 100-150 pl, or about 150-200 pl) if the subject is a human. In some examples, the amount of the modified peptide administered by intravitreal injection is about 2-20 pmoles of peptide (such as about 2-5, about 5-10, about 10-15, or about 15-20 pmoles of peptide) if the subject is a rodent (such as a mouse) or is about 1-10 pmoles of peptide (such as about 1-3, about 1.5-5, about 2.5-7.5, about 0.6-10, or about 1.5-7.5 pmoles of peptide if the subject is a human.

[0181] In other examples, the composition is administered to a subject with cancer or a CNS disease or disorder by any route, including intravenous, intraperitoneal, intramuscular, intraperitoneal, intrastemal, or intraarticular injection or infusion, or by transdermal, sublingual, oral, topical, intranasal, or transmucosal administration. In further examples, the composition is administered to a subject by local administration, such as administered to the CNS or to a tissue wherein a tumor is located, or directly into the tumor (e.g., intratumoral administration).

[0182] In some examples, the composition administered to a subject with cancer includes about 1 mg / kg to about 100 mg / kg of one or more modified peptides (such as about 1 mg / kg to about 10 mg / kg, about 5 mg / kg to about 25 mg / kg, about 20 mg / kg to about 50 mg / kg, about 40 mg / kg to about 80 mg / kg, about 50 mg / kg to about 75 mg / kg, or about 75 mg / kg to about 100 mg / kg). In some examples, the composition is provided in a unit dosage form, such as a selected quantity in a vial. In some examples, the unit dosage form includes about 25 mg to about 1g (such as about 25 mg to 50 mg, about 50 mg to 100 mg, about 100 mg to about 250 mg, about 200 mg to about 400 mg, about 250 mg to about 500 mg, about 300 mg to about 600 mg, about 500 mg to about 750 mg, or about 750 mg to about 1 g). One of ordinary skill in the art will recognize that these dosages can be varied, for example, as a result of clinical trials, the particular subject being treated, or other factors.

[0183] Compositions are formulated for injection generally by mixing a disclosed therapeutic agent at the desired degree of purity in a unit dosage injectable form (solution, suspension, or emulsion) with a pharmaceutically acceptable carrier, for example, one that is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation. Pharmaceutical compositions can include an effective amount of the peptide dispersed (for example, dissolved or suspended) in a pharmaceutically acceptable carrier or excipient. The nature of the carrier will depend on the particular mode of administration being employed. For example, formulations for injection usually contain injectable fluids that include pharmaceutically and physiologically acceptable fluids, such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol, or the like, as a vehicle. In addition, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, pH buffering agents and the like, for example, sodium acetate or sorbitan monolaurate. A disclosed therapeutic agent can be suspended in an aqueous carrier, for example, in water or an isotonic or hypotonic buffer solution at a pH of about 3.0 to about 8.5, such as about 4.0 to about 8.0, about 6.5 to about 8.5, or about 7.4. Useful buffers include buffered saline (such as phosphate-buffered saline or Hank’s balanced salt solution), artificial tears, or an ionic boric acid buffer. The active ingredient, optionally together with excipients, can also be in the form of a lyophilizate and can be made into a solution prior to administration by the addition of suitable solvents.

[0184] For solid compositions (such as powder, pill, tablet, or capsule forms), non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate. In addition to biologically neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate. To extend the time during which the peptide is available to inhibit or treat a retinal disease or disorder or a cancer, the peptide can be included in a liposome or as a particulate system. The particulate system can be a microparticle, a microcapsule, a microsphere, a nanoparticle, a nanocapsule, or similar particle. In some examples, the peptide is formulated as a long acting depot or sustained release microsphere. In some examples, the microsphere is a polymeric formulation, for example including but not limited to polylactide-co-glycolide (PLGA) or polylactic acid (PLA)- based microspheres (see, e.g., Butreddy et al., Int. J. Mol. Sci. 22:8884, 2021). Liposomes, including cationic and anionic liposomes, can be made using standard procedures as known to one skilled in the art. Liposomes including one or more of the disclosed peptides can be applied topically, either in the form of drops or as an aqueous based cream, or can be injected intraocularly. As one example, liposomes may be prepared from dipalmitoyl phosphatidylcholine (DPPC), such as egg phosphatidylcholine (PC). In a formulation for topical application, the peptide is slowly released over time as the liposome capsule degrades. In a formulation for intraocular injection, the liposome capsule degrades due to cellular digestion. Both of these formulations provide advantages of a slow release drug delivery system. In one example, the peptide can be dissolved in an organic solvent such as DMSO or alcohol and contain a poly anhydride, poly (glycolic) acid, poly(lactic) acid, or polycaprolactone polymer.

[0185] Pharmaceutical compositions for oral use can also be formulated, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion hard or soft capsules, or syrups or elixirs. Such compositions can be prepared according to standard methods known to the art for the manufacture of pharmaceutical compositions and may contain one or more agents selected from the group of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with suitable non-toxic pharmaceutically acceptable excipients including, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as com starch, or alginic acid; binding agents, such as starch, gelatin or acacia, and lubricating agents, such as magnesium stearate, stearic acid or talc. The tablets can be uncoated, or they may be coated by known techniques in order to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. Pharmaceutical compositions for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium such as peanut oil, liquid paraffin or olive oil.

[0186] In some examples, one or more PEDF peptides can be included in a delivery system that can be implanted at various sites in the eye, depending on the size, shape and formulation of the implant, and the type of transplant procedure. The delivery system is then introduced into the eye. Suitable sites include but are not limited to the anterior chamber, anterior segment, posterior chamber, posterior segment, vitreous cavity, suprachoroidal space, subconjunctiva, episcleral, intracorneal, epicorneal and sclera. In one example, the PEDF peptide delivery system is placed in the anterior chamber of the eye. In another example, the PEDF peptide delivery system is placed in the vitreous cavity.

[0187] The disclosed peptides can be administered by any means known to one of skill in the art, such as by intramuscular, subcutaneous, intraperitoneal, or intravenous injection, but even oral, nasal, or anal administration is contemplated. The disclosed peptides can also be administered topically, transdermally, or by local injection. In some embodiments, administration is orally, by intravenous injection, or subcutaneous injection. To extend the time during which the peptide is available, the peptide can be provided as an implant, an oily injection, or as a particulate system. The particulate system can be a microparticle, a microcapsule, a microsphere, a nanoparticle, a nanocapsule, or similar particle. In other examples, the peptide is present in an absorbable matrix, such as an absorbable polymer e.g., polylactide, polyglycolide, polylactide glycolide, etc.). One of ordinary skill in the art is aware of methods of administering peptides to a subject. See, e.g., Banga, “Parenteral Controlled Delivery of Therapeutic Peptides and Proteins,” in Therapeutic Peptides and Proteins, Technomic Publishing Co., Inc., Lancaster, PA, 1995. They may be administered in different forms, including but not limited to solutions, emulsions and suspensions, microspheres, particles, microparticles, nanoparticles, and liposomes.

[0188] The composition may be administered one or more times, such as twice daily, once daily, every other day, once weekly, every other week, every 4 weeks, every 6 weeks, or every 8 weeks. In some examples, the composition is administered topically to one or both eyes of the subject once daily or every other day. In other examples, the composition is administered every 2-6 weeks by intravitreal injection (such as every 2 weeks, every 4 weeks, or every 6 weeks). The subject may be treated for a set period of time or on a continuing basis. In some examples, the subject receives the treatment unless or until a therapeutic effect is no longer observed. A suitable treatment regimen, including frequency of administration and duration of treatment can be selected by a skilled clinician, depending on the disease or disorder being treated, the particular subject being treated, route of administration, and other factors. The disclosed peptides can be used alone or in combination therapy with other compositions or drugs used to treat the described conditions. One or more of the additional agents can be formulated separately, or in the same composition as the disclosed peptides. In some examples, the subject has AMD and is further administered ranibizumab, bevacizumab, pegaptanib, aflibercept, or pegcetacoplan. In other examples, the subject has a maculopathy and is further administered topical metformin. In further examples, the subject has PR or dry AMD and is further administered L1M21. In other examples, the subject has cancer, and is further administered one or more chemotherapeutic agents, immunotherapeutic agents, radiation therapy, and / or may be treated surgically. Additional agents that can be administered include antibacterial and / or antifungal antibiotics, as well as non-steroidal anti-inflammatory agents to reduce risk of infection and inflammation. Additional agents can be administered by any route, including topical administration or injection. A skilled clinician can select appropriate additional treatments, for example, based on the condition being treated and symptoms of the subject.

[0189] Efficacy of treatment can be evaluated by methods known to one of ordinary skill in the art, including measuring or assessing visual function, retinal function, or retinal structure. In some examples, methods for measuring or assessing visual function include a visual acuity test (such as using a Snellen chart, Allen picture cards, E chart, or Lebensohn chart, or ETDRS chart), color vision testing (such as Ishihara plates, Farnsworth-Munsell 100-hue test, Farnsworth D15 panel, or Lanthony desaturated test), visual field testing (such as confrontation field technique, tangent screen testing, automated perimetry device, or scanning laser ophthalmoscope), Amsler grid test, or visual sensitivity test (such as with a Goldman adaptometer). Retinal function can be measured indirectly (for example, utilizing the visual tests described above) or directly, for example by electroretinogram (ERG), such as full-field ERG, multi-focal ERG, S-cone ERG, focal ERG, or pattern ERG. Retinal function can also be assessed utilizing the electro-oculogram (EOG). In other examples, the structure of the retina is assessed, for example, utilizing fundus examination (for example, with direct or indirect ophthalmoscope or slit-lamp), ophthalmic ultrasonography, scanning laser ophthalmoscope, optical coherence tomography (OCT), or fluorescein angiography. In additional examples, the methods can include use of fluorescent dye (such as PSVue® 550) for evaluating retinal cell death or histology and immunofluorescent labeling of cell death markers (such as BAX and BCL2).

[0190] In some examples, treating or inhibiting a retinal disorder in a subject includes an increase in one or more measures of visual or retinal function (such as at least about a 20% increase, at least about a 50% increase, at least about a 75% increase, at least about an 80% increase, at least about a 90% increase, at least about a 1.5-fold increase, at least about a 2-fold increase, at least about a 3- fold increase, or at least about a 5-fold increase) in the subject as compared to a control. In other examples, treating a retinal disorder in a subject includes an increase or decrease (such as at least about a 20% increase, at least about a 50% increase, at least about a 75% increase, at least about an 80% increase, at least about a 90% increase, at least about a 1.5-fold increase, at least about a 2- fold increase, at least about a 3 -fold increase, or at least about a 5 -fold increase or at least about a 20% decrease, at least about a 50% decrease, at least about a 75% decrease, at least about an 80% decrease, or at least about a 90% decrease) in one or more measures of retinal structure (e.g., retinal thickness or number or size of lesions) in a subject as compared to a control.

[0191] The control can be any suitable control against which to compare visual function, retinal function, or retinal structure of a subject. In some embodiments, the control is a reference value or ranges of values. For example, the reference value can be derived from the average values obtained from a group of normal control subjects (for example, subjects without a retinal disorder). In further examples, the reference value is derived from the average values obtained from a group of subjects with a retinal disorder (such as the same or a different retinal disorder as the subject), for example, an untreated subject or a subject treated with vehicle alone. In other examples, the control is obtained from the same subject, for example, a subject with retinal disease prior to treatment.

[0192] EXAMPLES

[0193] The following examples are provided to illustrate particular features of certain aspects of the disclosure, but the scope of the claims should not be limited to those features exemplified.

[0194] Example 1 Peptide Synthesis

[0195] General: Fmoc-amino acids were procured from AAPTEC and Chem-Impex Inti. Biograde solvents were purchased from Oakwood Chemicals. Reagents used for couplings and deprotection were obtained from Chem-Impex Inti. Fmoc-serine preloaded Rink amide resin was purchased from AAPTEC. Automated synthesis of peptides were carried out on a AAPTEC Focus- 6V synthesizer employing standard Fmoc chemistry. LC / MS were recorded on a Agilent Technologies 1290 Infinity LC / XT MSD instrument.

[0196] Deprotection: Fmoc- group from amino acids were removed by shaking the resin with 20% piperidine in dimethylformamide (DMF) for 15 min (twice; 15.0 ml / g of the resin). The resin was drained, washed with DMF (three times; 15.0 ml / g of the resin) and was taken to the next step. Washing: The resin was drained from the deprotection / coupling mixture, and washed with DMF three times (15.0 ml / g) and after the final Fmoc -deprotection, the resin was washed with dichloromethane (DCM) three times

[0197] Coupling: 8.0 Equiv. of the amino acid to be coupled, was activated with 8.0 equiv. of 2- ( 1 H-benzotriazol- 1 -yl)- 1 , 1 ,3,3-tetramethyluronium hexafluorophosphate (HBTU), and 16.0 equiv. of 4-Methylmorpholine (NMM) in a mixing vessel, and thoroughly mixed for 2.0 min by bubbling nitrogen through the solution. This activated amino acid was then transferred to the resin in the reaction vessel under a positive pressure of nitrogen, and the vessel was shaken for 1 h while nitrogen gas was bubbled through the solution every 5 min. After 1 h of coupling time, the resin was drained, washed with DMF (thrice) and taken to the next deprotection and coupling steps until the sequence was completed on the solid phase.

[0198] Peptide cleavage from the resin: A cleavage cocktail of 98: 1 : 1 - trifluoroacetic acid (TFA):Triisopropylsilane (TIPS):water was added to the peptide vessel with the resin (15.0 ml / g) and shaken for Ih. The resin was filtered and washed with 2 x 10.0 ml of cleavage cocktail and the filtrates were combined and evaporated under reduced pressure at RT. The residue was triturated with anhydrous ether (20.0 ml) and decanted. This process was repeated two more times and the fully deprotected peptide residues were purified by preparative HPLC. The syntheses were started with 0.1 mmol of Rink amide resin pre-loaded with Fomc-Ser(t-Bu)-OH and / or Rink Amide resin. About 100-120.0 mg of the crude peptides obtained after the cleavage were taken to HPLC purification. Yields were in the range of 10 to 16%.

[0199] Preparative HPLC conditions: Column: X-Terra® (Waters corp.) C18 RP; 250 x 50 mm; 5.0 microns; solvent A: water with 0.1% TFA (v / v) and solvent B: acetonitrile with 0.1% TFA (v / v); Elution rate: 50.0 ml / min; Gradient: 10%B - 80%B over 70 min; Detection @ 220 nm. Fractions with the required mass and purity of >95% were pooled and freeze-dried to yield the peptides as colorless fluffy solids.

[0200] Analytical HPLC conditions A: Column: Zorbax® C18; 3,5 microns; 50 x 4.6 mm; Solvent A: Water with 0.1% TFA and B: acetonitrile with 0.1% TFA; Elution rate: 1.0 ml / min; 5% B - 95%B over 7 min; Detection @ 220nm.

[0201] Analytical HPLC conditions B: Column: Zorbax® Cl 8; 2.7 microns; 50 x 4.6 mm; Solvent A: Water with 0.1% TFA and B: acetonitrile with 0.1% TFA; Elution rate: 1.0 ml / min; 5% B - 95%B over 5 min; Detection @ 220nm. Table 2. LC / MS analysis of PEDF peptides

[0202] "MS data were obtained on a low resolution MS instrument, in contrast to the high resolution data for the other peptides.

[0203] Example 2

[0204] Effect of Modified PEDF peptides on Photoreceptor Cell Death

[0205] Animals were used following the Association for Research in Vision and Ophthalmology statement for the Use of Animals in Ophthalmic and Vision Research, USA ARVO guidelines. The mice were wild type C57BL / 6 J mice, rdlO (a mouse model for a human inherited autosomal recessive RP), and rdlO / Serpinfl null, a mouse without the Serpinfl gene in rd 10 background. Retinal degeneration in the rdlO mice is caused by a spontaneous mutation in the gene for the P- subunit of phosphodiesterase (Dixit et cd., Exp. Eye Res. 198: 108121, 2020).

[0206] Eyedrops (5 pl) of a solution of 1 mg / ml of modified PEDF 17mer peptides in HBSS were administered to the right eye (OD) and eyedrops of HBSS, without the peptide, to the left eye (OS) as control to rdlO and rdlO x Serpinfl null mice. Administration was daily to mice at ages between P15 and P20. PS Vue® was administered at age P20 to detect photoreceptor cell death at P21 by fluorescence fundoscopy. At end point (P21), eyes were enucleated and cross section of retinas from the treated animals were obtained. Histological evaluation of retinal cross sections was performed with hematoxylin and eosin staining.

[0207] The PSVue® assay detected cell death in eyes without the peptide but decreased with peptides 10 (SEQ ID NO: 12) and 11 (SEQ ID NO: 13) in rdlO mice (FIGS. 2A). Similar experiments were performed in rdlO / Serpinfl null mice (FIGS. 2B) with similar results. Peptide 14 (SEQ ID NO: 16) did not significantly decrease cell death in either mouse model (FIG. 3).

[0208] Histological evaluation of retina cross section stained with hematoxylin and eosin demonstrated that the outer nuclear layer (ONL) was thicker for eyes treated with eyedrops of peptide 10 (SEQ ID NO: 12) than without the peptide for both mouse models (FIGS. 5A-5B). Spider plots showed differences in ONL thickness in retinal areas spanning away from the optic nerve (ON) (FIGS. 5A-5B).

[0209] Similar experiments were earned out in rdlO mice with peptides Pl (SEQ ID NO: 3), Pl 8 (SEQ ID NO: 20), P19 (SEQ ID NO: 21), and P5 (SEQ ID NO: 7). Daily eyedrops of the indicated peptide (H2O was vehicle) were administered to rdlO mice. Dose was each eyedrop of 5 pl at 1 mg / ml of peptide in H2O in the right eye of each animal, and vehicle in the left eye between Pl 5- P20 (FIG. 6A). Then addition of eyedrops of PSVue® at P20, and the outcome was imaging of fluorescence fundoscopy 24 h after PSVue® eyedrops per eye at P21 using MICRON III. Three animals were treated with each of the peptides. Pl was the most efficient peptide among the peptides, followed by P19 and then P18, and P5 was inefficient (FIGS. 6B and 6C). Retinal histology was also performed at P21 and the thickness of the ONL was evaluated. Similarly to the fundoscopy results, Pl was the most efficient peptide among the peptides and Pl 8 and P19 performed similarly (FIGS. 6D and 6E).

[0210] Additionally, immunofluorescence of the retinal cross sections to detect pro-apoptotic BAX2 and anti-apoptotic BCL2 proteins showed a decrease of the pro-death marker and increase in the pro-survival marker in retinas from mouse eyes treated with the Pl, Pl 8 and P19 peptides relative to those with peptide P5 (FIG. 18A-18D). The findings suggest inhibition of apoptotic pathways and activation of survival mechanisms mediated by peptides Pl, P18, and P19 eye drop treatment. Together, the findings indicate that daily eye drops treatments with peptides Pl, Pl 8, and Pl 9 effectively protected rd 10 mice against photoreceptor cell death, demonstrating the potential of these peptides as a non-invasive approach for RP.

[0211] Example 3 Effect of PEDF Peptides on Phospholipase A Activity of PEDF Receptor Expression and purification of rhuPEDF-R(l-288): pGro7(groES-groEL), and pE-

[0212] SUMOstar-PEDF-R( 1-288) C-terminal TwinStrep tagged were co-transformed into Escherichia coli Rosetta(DE3) and plated on LB agar plate containing 10 pg / mL chloramphenicol and 100 pg / mL ampicillin followed by overnight incubation at 37°C. LB media (100 mL) containing 10 pg / mL chloramphenicol and 100 pg / mL ampicillin was inoculated with a single colony and grown overnight at 37°C with vigorous shaking. Then, 1 L LB media containing 0.5 mg / mL L (+) arabinose, 2% D-(+)-Glucose, 10 pg / mL chloramphenicol and 100 pg / mL ampicillin was inoculated with 2% of the overnight culture at 37°C with vigorous shaking until the O.D.600 reached 0.5-0.6. Protein expression was induced by addition of 0.5 mM IPTG at 25°C overnight and cells were harvested by centrifugation. The harvested cells were resuspended in buffer A (50 mM Tris- HC1, pH 8.0, 500 mM NaCl, 20% Glycerol, 1 mM TCEP, 0.1% IGEPAL® CA-630, 0.5% (W / V) CHAPS and 10 mM Imidazole) and EDTA-free protease inhibitor cocktail. Cells were lysed using a French press, and the lysate was cleared by centrifugation (33,745 g, 30 min, 4°C). The supernatant was used to purify the recombinant protein by affinity chromatography using a HisTrap™ HP column pre-equilibrated with 10 column- volumes of buffer A attached to an automated FPLC system (AKTA) with a flow rate 2 mL / min. The unbound proteins were washed with 10 column-volumes of buffer B (50 mM Tris-HCl, pH 8.0, 500 mM NaCl, 20% Glycerol, 1 mM TCEP, 0.1% IGEPAL® CA-630, 0.5% (W / V) CHAPS, 10 mM Imidazole, 10 mM ATP and 10 mM MgCh) and buffer A, respectively. Further the column was washed with 20 columnvolumes of buffer C (50 mM Tris-HCl, pH 8.0, 500 mM NaCl, 20% Glycerol, 1 mM TCEP, 0.1% IGEPAL® CA-630, 0.5% (W / V) CHAPS) containing 70 mM Imidazole). Bound proteins were eluted with 20 column-volumes of a linear gradient of 70 mM - 1000 mM imidazole in buffer C. Fractions containing proteins were pooled and loaded onto StrepTrap™ XT column preequilibrated with 10 column-volumes of buffer D (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% Glycerol, 1 mM TCEP and 0.5% (W / V) CHAPS) using an automated FPLC system (AKTA) with a flow rate 2 mL / min for further purification. The unbound proteins were washed with 20 columnvolumes of buffer D. Bound proteins were eluted with 20 column-volumes of buffer E (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% Glycerol, 1 mM TCEP, 0.5% (W / V) CHAPS and 100 mM Biotin). Fractions containing proteins were pooled, concentrated with Amicon Ultra (30K), and dialyzed against buffer F (10 mM HEPES, pH 7.5, 150 mM NaCl, 10% Glycerol, 0.5% (W / V) CHAPS and 1 mM TCEP). See flowchart with the purification protocol (FIG. 8A). The protein was >90% pure as determined by SDS-PAGE followed by Coomassie Blue staining. Protein concentration was determined by optical spectrometry (Nanodrop) using E280 nm = 31400 M1cm1.

[0213] Phospholipase activity assay: PLA activity assays were performed according to the manufacturer protocol (ThermoFisher Scientific). Briefly, rhuPEDF-R( 1-288) was mixed with IX phospholipase A reaction buffer to a concentration of 120 nM and then transferred to a microplate well. For testing the stimulation by PEDF peptide, rhuPEDF-R( 1-288) and PEDF derived peptides were incubated together at room temperature for 30 min and then diluted up to 50 pL with IX phospholipase A reaction buffer to make the final concentration of rhuPEDF-R( 1-288) 120 nM and PEDF derived peptides 500 pM, and transferred to microplate well. A lipid mixture was prepared by mixing together 15 pL 10 mM DOPC, 15 pL 10 mM DOPG and 15 pL 1 mM PLA substrate. Then, 2.5 mL substrate-liposome was prepared by mixing 25 pL lipid mixture with 2.5 mL IX phospholipase A reaction buffer by stirring. 50 pL substrate-liposome mixture was added to each microplate well containing rhuPEDF-R(l-288) with or without the PEDF derived peptide mixture, and incubated at room temperature for 10 minutes for PLA2 and 30 minutes for PLAi, protected from light. Then fluorescence emission was measured at 515 nm with excitation al 460 nm. Results: A nucleic acid encoding a truncated human PEDF-R (amino acids 1-288) was cloned into pE-SUMO star vector (FIG. 7). The rhuPEDF-R( 1-288) was expressed in bacteria and purified (FIGS. 8B and 8C).

[0214] To investigate whether the rhuPEDF-R [1-288] possesses both PLAi and PLA: activities, these individual enzymatic activities were assayed with specific substrates. First, to confirm the specificity of each substrate, control reactions were performed with a PLAi enzyme (Lecitase Ultra) that is free of PLA2 activity, and a PLA2 enzyme (honeybee venom) that is free of PLAi activity. Lecitase Ultra catalyzed only the PLAi substrate and not the PLA2 one (FIG. 9B). The PLA2 substrate was only hydrolyzed by honeybee venom PLA2 and not by Lecitase Ultra PLAi (FIG. 9C).

[0215] Second, concentration-dependent reactions with rhuPEDF-R[l-288] were performed and showed an increase in fluorescent intensity when reactions were with both substrates specific for PLAi and PLA2 (FIG. 9 A). These findings demonstrate that rhuPEDF-R[ 1-288] exhibits both PLAi and PLA2 activities.

[0216] Phospholipase A2 (PLA2) activity of the purified rhuPEDF-R( 1-288) was determined using a fluorescence-based assay (FIG. 13 A), confirming that purified rhuPEDF-R( 1-288) stimulated PLA2 activity. For comparison, the effect of PEDF 17mer peptide (SEQ ID NO: 1), PEDF H105A peptide (SEQ ID NO: 2), and PEDF R99A peptide (SEQ ID NO: 57) on PLA2activity of rhuPEDF- R(l-288) were tested (FIG. 10). The effect of modified PEDF peptides on PLA2 activity of rhuPEDF-R(l-288) was tested (FIG. 11). A dose-response experiment was carried out with selected peptides (FIG. 12).

[0217] Phospholipase Ai (PLAi) activity of the purified rhuPEDF-R( 1-288) was also determined using a fluorescence-based assay (FIG. 13B). The effect of modified PEDF peptides on PLAi activity of rhuPEDF-R( 1-288) was tested (FIG. 14). A dose-response experiment was also carried out with selected peptides (FIG. 15).

[0218] Additionally, the PLA enzymatic activity of rhuPEDF-R[ 1-288] was assessed in the presence of peptides P56 (SEQ ID NO: 61), P57 (SEQ ID NO: 62), P58 (SEQ ID NO: 63), and P3' (SEQ ID NO: 64). Among these, peptide P58 (SEQ ID NO: 63) uniquely and specifically stimulated PLAi activity. This finding suggests that P58 may play a role in promoting photoreceptor survival, a critical aspect for potential therapeutic applications (FIGS. 16A and 16B). Example 4

[0219] Effect of Additional Selected Peptides in rdlO Mice

[0220] Eyedrops (5 pl) of a solution of 1 mg / ml of each peptide P24 (SEQ ID NO: 26) and P31 (SEQ ID NO: 33) in H2O were administered to the right eye and of H2O alone to the left eye, as control, to rdlO mice. Administration was daily to rdlO mice at ages between P15 and P20 (see scheme in FIG. 6A). PSVue® was administered at age P20 to detect PS externalization, a first step in the cell death program of photoreceptors (FIG. 17A). PSVue® fluorescence in control eyes without peptide and in eyes with P31 detected PS externalization, but the fluorescence decreased in eyes that received peptide P24 indicating a decrease in PS externalization. (FIG. 17B). This demonstrates the benefit of peptide P24 eye drops in blocking PS externalization, an early step in the cell death process.

[0221] Early changes effected by RP can be histologically detected as shortening of photoreceptor OS and photoreceptor loss. Therefore, at end point (postnatal day 21), eyes were enucleated and cross sections of retinas from the animals were obtained for histological and immunofluorescence evaluation on morphology and markers of cell death pathway. First, histological evaluation of retina cross section stained with hematoxylin and eosin was performed. The retinas of rdlO mice treated with daily peptide 24 eye drops had longer outer segments and thicker outer nuclear layer (ONL) than vehicle-treated, while peptide 31 had no effect at postnatal day 21 (P21) (FIGS. 17C, 17D). Peptide 24 eye drops restored ONL thickness, while vehicle- and P31 -treated eyes were inefficient in restoring ONL thickness.

[0222] Example 5

[0223] Treating Cancer with Modified PEDF Peptides

[0224] The disclosed modified PEDF peptides have cytoprotective effects (for example as described in Example 2), which appear to be mediated by binding to the pigment epithelium- derived factor (PEDF) receptor, PEDF-R. PEDF has a number of biological effects including antiangiogenesis and decreasing inflammatory responses. The disclosed peptides are expected to have the anti-inflammatory response, but not the anti-angiogenesis effect. The biological response may also be due in part to stimulation of phospholipase A2 (PLA2) and / or PLAi (as demonstrated in Example 3). In addition to protective effects in eye disease, these peptides may be used in treating cancer. PEDF has been found in several cancers, including ovarian liver, pancreatic and skin (melanoma) cancers. The expression of PEDF may be associated with decreased metastasis and improved prognosis. Since PLA2 and PLAi lead to production of lysophosphatidylcholine (lysoPC), the finding that lysoPC has anti-tumor effects is also relevant. Also, higher plasma levels of lysoPC may lower the risk of cancer in a metabolomics study. The PLA2S that liberate arachidonic from phospholipids enriched in this fatty acid, which in turn has been shown to promote cancer and inflammation. The superfamily of PLA2S is likely responsible for these different effects.

[0225] It will be apparent that the precise details of the methods or compositions described may be varied or modified without departing from the spirit of the described aspects of the disclosure. We claim all such modifications and variations that fall within the scope and spirit of the claims below.

Claims

We claim:

1. A pigment epithelium-derived factor (PEDF) peptide comprising SEQ ID NO: 1 with one or more modifications, wherein the one or more modifications are selected from: a) an N-terminal modification; b) one or more amino acid substitutions; c) deletion of 1-4 amino acids from the N-terminus and / or C-terminus of the peptide; d) replacement of one or more amino acids with a non-alpha-amino acid linker; e) a C-terminal modification; and f) any combination of two or more thereof, wherein the peptide comprises 8-19 amino acids.

2. The PEDF peptide of claim 1 , wherein the peptide comprises or consists of any one of SEQ ID NOs: 3-54 or 61-64.

3. The PEDF peptide of claim 2, comprising or consisting of any one of SEQ ID NO: 3, SEQ ID NO: 26, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 12, and SEQ ID NO: 30.

4. The PEDF peptide of any one of claims 1 to 3, wherein the peptide comprises 8-17 amino acids.

5. The PEDF peptide of any one of claims 1 to 4, wherein the N-terminal modification comprises 6- aminohexanoic acid (AHA), aminopentanoic acid (APA), aminobutanoic acid (ABA), beta-alanine, or N-acetyl, or is derived from AHA, APA, ABA, or beta-alanine.

6. The PEDF peptide of any one of claims 1 to 5, wherein the one or more amino acid substitutions are at one or more positions corresponding to amino acid 5, 6, or 8 of SEQ ID NO: 1.

7. The PEDF peptide of claim 6, wherein the one or more amino acid substitutions comprise threonine, aspartic acid, diaminopropionic acid (DAP), or CF lhreonine at a position corresponding to amino acid 5 of SEQ ID NO: 1.

8. The PEDF peptide of claim 6 or claim 7, wherein the one or more amino acid substitutions comprise norleucine, leucine, valine, or N-methylated isoleucine at a position corresponding to amino acid 6 of SEQ ID NO: 1.

9. The PEDF peptide of any one of claims 6 to 8 wherein the one or more amino acid substitutions comprise alanine at a position corresponding to amino acid 8 of SEQ ID NO: 1.

10. The PEDF peptide of any one of claims 1 to 9, wherein the peptide comprises deletion of 1- 4 amino acids from the N-terminus of the peptide, deletion of 1-4 amino acids from the C-terminus of the peptide, or both.

11. The PEDF peptide of any one of claims 1 to 10, wherein the non- alpha- amino acid linker is 6-aminohexanoic acid (AHA), 5 -aminopentanoic acid (APA), 8-amino-3,6,dioxaoctanoic acid (AOC), 4’-amino-[l,l’-biphenyl]-4-carboxylate (4’ABPC), or 4-(4-(aminomethyl)phenoxy)benzoic acid (4’AMPC), 8-aminooctanoic acid (AOA), or is derived from AHA, APA, AOC, 4’ABPC, 4’AMPC, or AOA.

12. The PEDF peptide of claim 11, wherein the peptide comprises substitution of the amino acid corresponding to position 8, 9, or 10 of SEQ ID NO: 1 with the non-alpha-amino acid linker.

13. The PEDF peptide of claim 11 or claim 12, further comprising deletion of 1-3 amino acids following the position of substitution with the non-alpha-amino acid linker.

14. The PEDF peptide of any one of claims 1 to 13, wherein the C-terminal modification comprises C-terminal amidation, benzylamine-NH2, NH2-ethyl, or NH2CH2CH2OH.

15. The PEDF peptide of any one of claims 1 to 14, wherein the PEDF peptide increases phospholipase Ai activity of PEDF-R(l-288), phospholipase A2 activity of PEDF-R( 1-288), or both, compared to a control.

16. A composition comprising one or more PEDF peptide of any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

17. The composition of claim 16, wherein the pharmaceutically acceptable carrier comprises buffered saline or water.

18. The composition of claim 17, wherein the buffered saline is Hank’s balanced salt solution.

19. The composition of any one of claims 16 to 18, wherein the composition is formulated as an eye drop.

20. The composition of any one of claims 16 to 19, wherein the composition is formulated for injection.

21. The composition of any one of claims 16 to 20, wherein the composition comprises about 1 mg / ml of the peptide.

22. A method of treating a retinal disease or disorder, comprising administering the composition of any one of claims 16 to 21 to an eye of a subject with the retinal disease or disorder.

23. The method of claim 22, wherein the composition is administered topically to the eye of the subject.

24. The method of claim 22, wherein the composition is administered to the eye of the subject by injection.

25. The method of any one of claims 22 to 24, wherein the composition is administered to the eye of the subject twice daily, once daily, every other day, or every three days.

26. The method of any one of claims 22 to 26, wherein the retinal disease or disorder is retinal degeneration, macular degeneration, or glaucoma.

27. The method of claim 26, wherein the retinal degeneration is retinitis pigmentosa, Leber congenital amaurosis, or cone-rod dystrophy.

28. The method of claim 26, wherein the macular degeneration is age-related macular degeneration, Stargardt- like macular degeneration, vitelliform macular dystrophy (Best disease), bull’s eye maculopathy, or other maculopathy.

29. A method of treating or inhibiting cancer in a subject, comprising administering the composition of any one of claims 16 to 18 or 20 to the subject with the cancer.

30. The method of claim 29, wherein the composition is administered to the subject by injection or infusion or orally.

31. The method of claim 29 or claim 30, wherein the cancer is ovarian cancer, pancreatic cancer, liver cancer, or melanoma.

32. A method of treating or inhibiting a central nervous system (CNS) disease or disorder in a subject, comprising administering the composition of any one of claims 16 to 18 or 20 to the subject with the CNS disease or disorder.

33. The method of claim 32, wherein the CNS disease or disorder comprises cell death in the CNS.

34. The method of claim 32 or 33, wherein the CNS disease or disorder is amyotrophic lateral sclerosis, Huntington’s disease, Alzheimer’s disease, Parkinson’s disease, human immunodeficiency virus-induced encephalitis, or CNS trauma.

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