Serpin peptides and methods of use thereof

Abstract

Disclosed herein are serpin peptides, analogs and derivatives thereof, and their uses for treating various conditions associated with LRP1 or TSLP. TIFF2025503607000041.tif85128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of, and priority to, U.S. Provisional Patent Application No. 63 / 266,444, filed January 5, 2022, the disclosure of which is incorporated by reference in its entirety herein.

[0002] Sequence Listing This application contains an ST.26 compliant Sequence Listing, which has been filed contemporaneously with this application through the Patent Center in .xml format and is incorporated herein by reference in its entirety. The .xml copy created on December 28, 2022 is named Serpin Pharma 138536-8001WO01 Sequence Listing.xml and is 43.3 KB in size. [Background technology]

[0003] background Serine protease inhibitors (SERPINs) are a large family of proteins involved in diverse biological functions such as fibrinolysis, blood coagulation, and inflammation. When serpins bind to their target serine proteases and inactivate their enzymatic activity, a conformational change occurs, exposing a unique short peptide motif (5-11 amino acids) that 8,43 The protease-inhibitor complex binds to the low density lipoprotein receptor-related protein (LRP1) at a newly exposed short peptide motif, a process that is conserved across the range of serpins, such as alpha-1 antitrypsin (AAT) and antithrombin III (ATIII) (May 2013) 22,25,43 Therefore, there is a need to develop novel serpin peptides and explore their preventive and therapeutic effects in various conditions and diseases. Summary of the Invention

[0004] overview In some embodiments, the present technology provides a method for administering to a subject a therapeutically effective amount of a medicament for reducing inflammation associated with a disease or condition associated with LRP1 or TSLP. The present invention relates to a method of reducing inflammation, including but not limited to innate immunity, adaptive immunity, eosinophilic inflammation, allergy, rhinitis, asthma, dermatitis, esophageal eosinophilia, eosinophilic asthma, atopic dermatitis, nasal polyps, pruritus, chronic idiopathic urticaria, and the use of serpin peptides in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP, comprising administering a serpin peptide selected from the group consisting of TIFF2025503607000002.tif48164. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 35 or SEQ ID NO: 2. In some aspects, the N-terminus of the serpin peptide is acetylated. In some aspects, the C-terminus of the serpin peptide is amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In some aspects, the other peptide is different from the serpin peptide. In aspects, the fusion peptide or fusion protein comprises a serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human and the serpin peptide is administered orally, parenterally, intradermally, transdermally, topically, or intranasally. In some aspects, the serpin peptide is administered as a single dose.

[0005] In some aspects, the disease or condition is caused by A. alternata. In some aspects, the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia. In some aspects, the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain. In some aspects, the disease or condition is an eosinophil-mediated disease (EDD), such as eosinophilic esophagitis (EoE), eosinophilic asthma, atopic dermatitis, nasal polyps, or chronic idiopathic urticaria. In some aspects, the disease or condition is atopic dermatitis or pruritus, and the serpin peptide can be administered by topical administration. In some aspects, the disease or condition is an allergic reaction, allergic inflammation, or eosinophil-mediated allergic disease.

[0006] In other embodiments, the present technology provides administering to a subject, The present invention relates to a method of treating or use of a serpin peptide in treating a subject having a disease or condition associated with LRP1 or TSLP, comprising administering a serpin peptide selected from the group consisting of: TIFF2025503607000003.tif41165, wherein the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain. In an aspect, the serpin peptide comprises the sequence of SEQ ID NO: 35. In an aspect, the N-terminus of the serpin peptide is acetylated. In an aspect, the C-terminus of the serpin peptide is amidated. In an aspect, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In an aspect, the other peptide is different from the serpin peptide. In an aspect, the fusion peptide or fusion protein comprises a serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender.

[0007] In aspects, the subject is a human and the serpin peptide may be administered orally, parenterally, intradermally, transdermally, topically, or intranasally. In aspects, the serpin peptide is administered to the subject at a dose of 0.001 mg / kg to 5 mg / kg, and the serpin peptide may be administered as a single dose. In aspects, administration of the serpin peptide results in relief of pain and / or administration prevents or reduces the onset of pain.

[0008] In some embodiments, the present technology provides administering to a subject, The present invention relates to a method of treating or use of a serpin peptide in treating a subject having a disease or condition associated with LRP1 or TSLP, comprising administering a serpin peptide comprising an amino acid sequence selected from the group consisting of TIFF2025503607000004.tif48165, wherein the disease or condition is caused by A. alternata. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 35, the N-terminus of the serpin peptide may be acetylated and / or the C-terminus of the serpin peptide may be amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein, and the other peptides may be different from the serpin peptide. In some aspects, the fusion peptide or fusion protein comprises a serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender.

[0009] In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg, and the subject can be a human. In some aspects, administration is by oral, parenteral, intradermal, transdermal, topical, or intranasal administration, and the serpin peptide can be administered as a single dose. In aspects, the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia, and administering the serpin peptide can reduce inflammation or eosinophilic inflammation. [Brief description of the drawings]

[0010] [Figure 1] We demonstrate that serpins contain an anti-inflammatory core motif that includes the LRP1-binding sequence. In LPS-stimulated NFκB reporter assays, the NFκB-lowering activity of the coreserpin peptide was lost when the truncation was too short, due to instability of the truncated peptide. Activity was restored by polyarginine residues that stabilized the peptide. However, when the LRP1-binding site was truncated, activity could not be rescued by polyarginine flanks. [Figure 2A]Figures 2A-2D show that SP163M promoted neurite length and growth associated protein-43 (GAP-43) in adult primary rat DRG neurons. Figure 2A: Representative phase contrast images of cultured primary adult DRG neurons over time. Cultures were treated daily with vehicle or SP163M (100 ng ml) for 48, 72 and 96 h. Scale bar 500 μm. Figure 2B: Representative images of immunofluorescence to detect βIII-tubulin in primary cultured adult DRG neurons in control and SP163M (240 nM) treated cells after 54 h. Top panel: Scale bar 200 μm; bottom panel: Scale bar 50 μm. Note the greater neurite length in SP163M treated neurons. Figure 2C: Quantification of immunofluorescence analysis in control (n = 144 neurons) and SP163M (n = 222 neurons) samples from 11 different cultures. SP163M significantly increased maximum neurite length compared to control. Mann-Whitney test, rank sum of vehicle-treated and SP163M-treated: 23537,43625; **p < 0.01. Data are presented as mean ± SEM. Figure 2D: Primary cultured adult DRG neurons were treated with vehicle or SP163M (240 nM) for 24 h. RT-qPCR analysis of GAP-43 mRNA levels (n = 8 independent studies). SP163M significantly increased regeneration-related genes compared to vehicle. Mann-Whitney test, rank sum of vehicle or SP163M: 29,76, **p < 0.01. [Figure 2B] See legend to Figure 2A. [Figure 2C] See legend to Figure 2A. [Figure 2D] See legend to Figure 2A. [Figure 3A]Figures 3A-3F show that SP163M activated transient cell signaling in an LRP1-dependent manner in PC12 cells. Figure 3A: Dose-dependent (0-240 nM) activation of phospho-ERK1 / 2 by SP163M after 10 min. Figure 3B: Time course (0-30 min) of activation of phospho-ERK1 / 2 by SP163M (240 nM). The last lane (far right) shows activation of phospho-ERK1 / 2 by the known LRP1 agonist EI-tPA (12 nM). Equal amounts of protein lysate (20 μg) were loaded per lane. Immunoblot analysis detects phospho-ERK1 / 2 and total ERK1 / 2 as loading controls. Figure 3C: RT-qPCR analysis of LRP1 mRNA after 48 h transfection of LRP1 siRNA. Data are presented as mean ± SEM; n = 3 independent experiments. T-test, T = 8.024, df = 4, **p < 0.05. Figure 3D: Immunoblot of LRP1 levels in PC12 cells transfected with non-targeting control (NTC) or siLRP1 for 48 h in PC12 cells. Figure 3E: Representative immunoblot of phospho-ERK1 / 2 activated by SP16 (240 nM) over time 48 h after transfection with NTC or siLRP1. Figure 3F: Immunoblot showing activation of phospho-Akt and phospho-ERK1 / 2 following pretreatment with vehicle or SP163M (24 or 240 nM) for 10 min and, in some wells, RAP (150 nM) for 15 min. NGF (0.36 nM) for 10 min served as a cell signaling control. Equal amounts of protein lysate (20 μg) were loaded per lane. Total ERK served as a loading control. [Figure 3B] See legend to Figure 3A. [Figure 3C] See legend to Figure 3A. [Figure 3D] See legend to Figure 3A. [Figure 3E] See legend to Figure 3A. [Figure 3F] See legend to Figure 3A. [Figure 4]Figures 4A-4C show that SP163M modulated both the early and late phases of the formalin test. Figure 4A: Time course of formalin (2.5%)-induced paw licking in C57BL6 mice. Vehicle or SP163M (0-2 μg / g sc) was administered 1 h before formalin injection into the hind paw. Figure 4B: Quantification of the area under the curve (AUC) of vehicle and SP163M (0.02, 0.2 and 2 μg / g) during the early phase. Data are presented as mean ± SEM; one-way ANOVA, F = 9.523, ***p < 0.001. Tukey's post hoc test, vehicle (n = 9) vs. SP163M 0.02 μg / g (n = 8), *p < 0.05; vehicle vs. SP16 0.2 μg / g (n = 8), ***p < 0.001; vehicle vs. SP163M 2.0 μg / g (n = 8), **p < 0.01. Figure 4C: Quantification of the area under the curve (AUC) of vehicle and SP163M (0.02, 0.2 and 2 μg / g) during the late phase. Data are presented as mean ± SEM; one-way ANOVA, F = 10.2, ****p < 0.0001. Tukey's post-hoc test, vehicle (n = 9) vs. SP163M 0.02 μg / g (n = 8), p = ns; vehicle vs. SP16 0. 2 μg / g (n = 9), ***p < 0.005; vehicle vs. SP163M 2.0 μg / g (n = 8), ***p < 0.005. [Diagram 5]Figures 5A-5C show that systemically administered SP163M and EI-tPA attenuated acute nociception induced by intraplantar capsaicin. Figure 5A: Nociception-related behavior (time spent licking) was increased in both male and female mice after intraplantar injection of capsaicin (20 μg) compared to vehicle (cyclodextrin: 20%). Data are presented as mean ± SEM. Kruskal-Wallis test ****p < 0.0001. Dunn's multiple comparison test post hoc test Male vs. female no capsaicin (n = 6), p = ns; Male vs. female with capsaicin (n = 8), p = ns; Male no capsaicin vs. male with capsaicin **p < 0.01; Female no capsaicin vs. female with capsaicin *p < 0.5. Fig. 5B: Nociception-related behavior (time spent licking) after administration of LRP1 interactors, SP163M (2 μg / g) or EI-tPA (2 μg / g) in male mice with intraplantar capsaicin for 10 min. Data are means ± SEM. One-way ANOVA, F = 21.28 ***p < 0.001. Tukey's post hoc test, Vehicle (n = 13) vs. SP163M (n = 8), **p < 0.01; Vehicle vs. EI-tPA (n = 6), **p < 0.01; Fig. 5C: Nociception-related behavior (time spent licking) after administration of LRP1 interactors, SP163M (2 μg / g) or EI-tPA (2 μg / g) in female mice with intraplantar capsaicin for 10 min. Data are means ± SEM. One-way ANOVA, F = 14.20 ***p < 0.005. Tukey's post-hoc test, vehicle (n = 13) vs. SP16 (n = 8), ***p < 0.005; vehicle vs. EI-tPA (n = 6), **p < 0.01. [Figure 6A]Figures 6A-6E show that systemically administered SP163M treatment blocked the development of mechanical hypersensitivity and recruitment of inflammatory cells after PNL. Figure 6A: Tactile allodynia developed after PNL and persisted for 14 days. Daily subcutaneous delivery of SP163M (2 μg / g) significantly prevented the development of tactile allodynia for 9 days after injury (**p < 0.01). Data are presented as mean ± SEM (n = 7 mice / group). Two-way ANOVA, days (main effect) F(6,42) = 2.51, *p < 0.05; treatment (main effect) F(1,42) = 57.91, ****p < 0.0001; treatment × days (interaction) F(6,42) = 0.7512 p = ns. Sidak's multiple comparison test Vehicle vs. SP16 post-injury, day 2 **p < 0.01, day 4 **p < 0.01, day 7 *p < 0.05, day 9 *p < 0.05. Figures 6B and 6D: Immunoblot analysis of injured sciatic nerves 2 days after PNL in vehicle- and SP163M-treated mice. SP163M treatment reduced inflammatory cell infiltration (CD11b) and suppressed TLR4. Figure 6C: Densitometric analysis of CD11b. One-way ANOVA, F = 39.05; ****p < 0.0001, Tukey's post-hoc test, control (n = 5) vs. vehicle (n = 6) ***p < 0.005, vehicle vs. SP16 (n = 5) **p < 0.01. Figure 6E: Densitometric analysis of TLR4. One-way ANOVA, F = 18.54; ****p < 0.0001, Tukey's post-hoc test, control (n = 6) vs vehicle (n = 6), ***p < 0.005 vehicle vs SP163M (n = 6) ***p < 0.005. All data are presented as mean ± SEM. [Figure 6B] See legend to Figure 6A. [Figure 6C] See legend to Figure 6A. [Figure 6D] See legend to Figure 6A. [Figure 6E] See legend to Figure 6A. [Figure 7]Figures 7A-7C show that SP163M reduced recruitment of inflammatory cells and activation of satellite cells after PNL. Figure 7A: Transverse sections of L4 DRG immunostained for CD11b and GFAP after treatment with vehicle or SP163M 2 days after PNL. Note abundant immunoreactivity identifying CD11b (black arrows, upper panel) between and near blood vessels or GFAP (black arrows, lower panel) around neuronal cell bodies in injured vehicle-treated DRG (left) or SP163M-treated DRG (right). CD11b and GFAP immunoreactivity in SP163M-treated DRG is minimal. Nuclei are stained with hematoxylin (blue). Figure 7B: Quantification of CD11b in DRG. Mann-Whitney test, rank sum for vehicle-treated and SP163M-treated: 23537, 43625; **p < 0.01. Data are presented as mean ± SEM. Mann-Whitney U test, rank sum for vehicle (n = 8) and SP163M (n = 6): 77, 28; *p < 0.05. Figure 7C: Quantification of GFAP in DRG. Mann-Whitney U test, rank sum for vehicle (n = 7) and SP163M (n = 9): 91, 45; ***p < 0.005. [Figure 8A] Figures 8A-8C show that SP163M blocked IL-13-stimulated phosphorylation of Stat6. Figure 8A: 30 min after IL-13 induction, STAT6 was phosphorylated in vehicle and A1AT-treated cells, but not in SP163M-treated cells. Figure 8B: This reduction in STAT6 phosphorylation by SP163M persisted for several hours after treatment (many different experimental replicates shown). Figure 8C: Shows that the reduction in phosphorylated STAT6 by SP163M was dependent on the expression of LRP1. Using CRISPER / CAS9 technology, we generated an LRP1 knockout esophageal EPC2 cell line. In control cells (with LRP1), SP163M was able to reduce phospho-STAT6 expression, but in the LRP1 knockout cell line, SP163M failed to reduce phosphorylated STAT6. [Figure 8B]See legend to Figure 8A. [Figure 8C] See legend to Figure 8A. [Figure 9] Figures 9A-9B show that SP163M reduced eosinophilic esophagitis in an eosinophilic esophagitis model using A. alternata as an allergen. Figure 9A: Representative images of anti-MBP staining of esophageal sections after allergen (A. alternata) or control (saline) challenge. Figure 9B: Quantification of eosinophils in the esophagus; data shown are eosinophil counts per high power field (HPF). [Figure 10A] Figures 10A-10C show that SP163M inhibited TSLP and reduced cell death in human keratinocytes. Figure 10A: HaCat cells were left untreated or induced with PolyIC (50 μg / ml) for 24 hours. Cells were pretreated with either vehicle or SP163M (100 μg / ml). TSLP was measured in the supernatant by ELISA (p = 0.0045 vs. vehicle-treated PolyIC-induced cells). Figure 10B: HaCat cells treated as in Figure 10A were analyzed by Promega's CellTiter-Glo® Luminescent Cell Viability Assay. Percent change from untreated (no PolyIC) cells was compared between SP163M vs. vehicle-treated PolyIC-induced controls (p = 0.0068). (C) HaCat cells were injured with TNFα and immunoblotted for phospho-IkBa (Ser32). GAPDH served as a loading control and signal normalization. [Figure 10B] See legend to Figure 10A. [Figure 10C] See legend to Figure 10A. [Figure 11]We show that SP163M improved the therapeutic outcome in an animal model of atopic dermatitis. Dermatitis was induced in the skin of BALB / c mice by a series of calcipotriol (MC903) and OVA challenges over an 8-week period. SP163M (100 μg / mouse), A1AT (2 mg / mouse) or vehicle (DDW) were administered twice weekly starting at week 4. The number of eosinophils infiltrating the epithelial sections was quantified by MBP staining. [Figure 12] Figures 12A-12B show that SP16 inhibits key allergic inflammatory mediators. Figure 12A: Poly I:C-induced TSLP production by SPINK7 knockout EPC2 cells (human esophageal epithelial cells). SPINK7 KO cells and control cells were plated at high calcium and high density for 48 hours, and then treated with SP16 or 7G and poly I:C (5 μg / ml, or untreated) for 8 hours. TSLP production was measured in the supernatant by ELISA. Figure 12B: Primary esophageal epithelial cells (EPC2) were treated with SP16 and then stimulated with IL-13, a TH2 response-mediating cytokine, to induce CCL26. SP16 significantly reduced CCL26 release in a dose-dependent manner. [Figure 13] Figures 13A-13C show that SP16 reduces inflammatory markers in bronchoalveolar lavage fluid of mice after OVA challenge. Mice were subjected to an OVA challenge allergic inflammation model and treated with vehicle or SP16 during the challenge phase. Bronchoalveolar lavage fluid (BALF) was analyzed for cytokines by ELISA. Figure 13A: SP16 significantly reduces CCL24 after OVA challenge in sensitized mice. Figure 13B: SP16 significantly reduces IL-1a after OVA challenge in sensitized mice. Figure 13C: SP16 significantly reduces IL-2 after OVA challenge in sensitized mice. [Figure 14]We show that SP16 treatment mediates TH2-type inflammatory cytokines in the OVA model. Mice were subjected to an OVA-challenged allergic inflammation model and treated with vehicle (saline), dexamethasone (1 mg / kg) (positive control) or SP16 (50 μg) for a total of four treatments during the challenge phase (intranasal OVA, four treatments over eight days). BALF and lung tissue were analyzed for cytokines by ELISA. Cells in BALF were counted with a hemocytometer, and total WBC counts were presented. [Figure 15A] Figures 15A-15B show that SP16 reduces eosinophil infiltration into the lung in the OVA model. Figure 15A: Mice were subjected to an OVA-challenged allergic inflammation model and treated with vehicle, SP16 or A1AT during the challenge phase. BALF was collected and analyzed for the percentage of eosinophils and compared with saline (unchallenged) mice, vehicle OVA-induced mice or SP16-treated OVA-induced mice. Figure 15B: Results of flow cytometry analysis of the BALF in Figure 15A are shown. [Figure 15B] See legend to Figure 15A. [Figure 16A] Figures 16A-16C show that SP16 improves treatment outcome in an AD-like inflammatory skin model. SP16 (in 70% ethanol) was applied topically to the ear on the day of MC903 application (MC903 + SP16). Control animals (MC903) received drug vehicle (70% EtOH) and normal control animals (NC) received EtOH instead of MC903. In-life parameters: Dermatitis score (Figure 16A) and ear volume (Figure 16B) were measured on days 0, 3, 7, 9, 11 and 14. As shown in Figure 16C, photos were taken on the last day of the study (day 15) and show a randomly selected representative mouse from each group. Clinical dermatitis scores (score 0-4 for each parameter) - erythema, scaling / dryness, edema, abrasions / erosions (total score 0-16). [Figure 16B] See legend to Figure 16A. [Figure 16C] See legend to Figure 16A. [Figure 17-1]FIG. 17 shows that SP16 treatment results in attenuation of pruritus through PAR2 and TSLP inhibition: AD was induced in mice using daily MC903 applications. Mice were treated with SP16 (MC903 + SP16) or drug vehicle (MC903) in 70% EtOH. Normal control (NC) mice were treated with EtOH only. Ears were measured for PAR2 protein expression by Western blot analysis (quantitative analysis after normalization to β-actin is shown) (n=3-5). TSLP was measured in both ears and serum by ELISA (N=3-5). Scratching behavior was assessed over a 3-min period after a 5-min acclimation period and the number of scratches was shown (n=3). IL-4 was measured in ear lysates by ELISA (N=3-5). [Figure 17-2] See description of Figure 17-1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] Detailed Description Disclosed herein are C-terminal peptides derived from serpin molecules, their variants and derivatives, and their use in preventing or treating various conditions by targeting LRP1.In certain embodiments, the serpin peptide is an isolated peptide.In certain embodiments, the serpin peptide is a synthetic peptide.

[0012] Also disclosed herein are pharmaceutical compositions comprising peptides derived from serpins, and methods of using same to treat a number of conditions, such as conditions associated with peripheral nerve injury (and resulting pain), and allergic inflammation, in which dysregulation of immune responses or impaired endocytic function, or mediation of LRP1, may contribute to the pathology. The unexpected regenerative and healing properties of these peptides allow compositions comprising such peptides to be used in new indications, such as preventative intervention in conditions associated with tissue damage.

[0013] LRP1-Related Conditions LRP1 functions as an endocytic and cell signaling receptor and has several ligands that induce specific cell signaling cascades that may contribute to cell survival and anti-inflammatory mechanisms. 5,18,22,25 LRP1 is ubiquitously expressed in many different organs and is abundant in the brain, lung, heart and immune cells. Due to these unique capabilities and its widespread expression in both tissues and immune cells, LRP1 plays a key role in inflammation, regulating cellular metabolism, and maintaining homeostasis. For example, LRP1 regulates inflammatory signaling pathways such as the NFκB and JNK pathways, which induce the conversion of pro-inflammatory (M1) macrophages to an anti-inflammatory (M2) macrophage phenotype, modulates cytokine production, and contributes to effective migration and phagocytosis. 22,26,51 In neutrophils, LRP1-dependent mechanisms lead to enhanced cell adhesion, chemotaxis, and antimicrobial activity of these cells, thereby resisting immunosuppression. 25 During acute infection or injury, LRP1 also promotes the resolution of inflammation and prevents the tissue injury cycle by clearing PAMPS and DAMPS from dying or damaged tissues. 25 LRP1 has also been shown to mediate autophagy during infection, a key metabolic process that has recently been shown to play an important protective role in various diseases. 4,10 Therefore, given its multifunctional ability to regulate inflammation, targeting LRP1 offers considerable potential to alleviate several aspects of the immune response that contribute to the pathology of several diseases, including neurological disorders, infectious diseases, and allergic inflammatory disorders.

[0014] Neurological disorders Peripheral nerve injury (PNI) resulting from metabolism, chemotherapy, or trauma often results in chronic pain. Neuropathic pain is characterized by evoked (allodynia, hyperalgesia) and spontaneous pain-like symptoms. Symptoms can be severe, including paresthesias, tingling, numbness, and burning sensations. Few treatment options are available beyond short-term symptomatic relief, including steroids, local anesthetics, antidepressants, anticonvulsants, and opioids, which are used only for severe pain. All of these treatments aim to temporarily reduce pain to manageable levels, but all of them can cause side effects and addiction, and do not promote healing of damaged nerves. Thus, there is an unmet clinical need for novel and innovative pain treatments to prevent the transition from acute to chronic pain.

[0015] Regarding nerve injury and associated pain, injury to the peripheral nervous system induces increased expression of LRP1. Previously, it has been demonstrated that LRP1 agonists can promote axonal growth in the CNS and induce regeneration after spinal cord injury. 53 LRP1 is an endocytic receptor for a diverse number of ligands, including tissue-type plasminogen activator (tPA), matrix metalloproteinase-9 (MMP-9), and activated α2-macroglobulin. 14 These ligands induce anti-inflammatory activity. 39 , activating the repair program of Schwann cells 21 , which can transactivate cell signaling pathways in neurons related to axon regeneration 38 Although LRP1 requires ligand binding to activate cell signaling, different ligands induce different, and sometimes opposing, cell signaling responses, reflecting the ability of different ligands to assemble unique co-receptor complexes. Furthermore, many LRP1 ligands are multi-domain proteins that exert various effects on cell physiology that are not involved in LRP1 binding. For example, tissue-type plasminogen activator (tPA) binds to LRP1 to promote Schwann cell (SC) survival and migration. 23However, tPA induces pain through LRP1-independent activity. 1 EI-tPA promotes the survival of human iPSC-derived neural progenitor cells (iNPCs), and transplantation of EI-tPA-activated iNPCs into rodents with severe spinal cord injury has been shown to improve recovery of motor function. 40 Microenvironmental imbalance following nerve injury can have serious consequences, including the development of chronic neuropathic pain states. 12 In peripheral nervous system (PNS) injury, both proinflammatory cytokines, such as TNFα, IL-6, and IL-1β, and anti-inflammatory cytokines, such as IL-10, have been shown to play a central role in axonal regeneration and repair. 6 .

[0016] Eosinophilic Disorders (EDD) Eosinophilic disorders (EDDs) are disorders associated with a type 2 inflammatory response with elevated eosinophil levels and eosinophil-mediated immune dysfunction. EDDs include EoE, eosinophilic asthma, atopic dermatitis, nasal polyposis, and chronic idiopathic urticaria.

[0017] The peptides disclosed herein may serve as adjunctive treatments to alleviate acute inflammatory responses in patients with ST-segment elevation myocardial infarction (STEMI). Furthermore, SP163M has been shown to be safe and well tolerated in both Phase I and ongoing Phase IIa clinical trials. This peptide therapy is unique in that it rebalances dysregulated immune responses and protects tissues from damage without significant immunosuppressive effects. It is therefore a safe anti-inflammatory drug with broad utility across a wide variety of immune-mediated diseases.

[0018] EoE is a chronic, predominantly type 2 immune-mediated, allergic inflammatory response associated with esophageal dysfunction and epithelial barrier dysfunction. This food allergen-driven disease is characterized by a type 2-mediated immune response that leads to eosinophil-dominant inflammation and esophageal damage. An estimated 150,000 patients (mostly children) in the United States currently suffer from this disorder, which often causes esophageal pain, difficulty swallowing, food impaction, persistent heartburn, chest and abdominal pain, weight loss, nausea, vomiting, and failure to thrive. Currently, there are no therapeutic agents approved by the FDA for the treatment of EoE, and management of the disease consists of dietary restrictions, proton pump inhibitors, and corticosteroids. The current standard of care does not adequately address the immune dysregulation that occurs in EoE and other allergic inflammatory diseases such as asthma or atopic dermatitis.

[0019] EoE is characterized by high numbers of eosinophils, proteases, cathelicidin, serine proteases including kallikrein (KLK5), and thymic stromal lymphopoietin (TSLP), a cytokine and master regulator of allergic type 2 inflammatory responses in the local environment. 41 In esophageal epithelial cells, loss of function of the serine peptidase inhibitor Kazal type 7-SPINK7 leads to uncontrolled protease activity and release of proinflammatory cytokines, such as TNFα, CCL2, GM-CSF, IL-8 and CXCL10, resulting in inflammation. Recently, it has been shown that the serine protease KLK-5, a key mediator of epithelial barrier function, is a direct target of SPINK7, and loss of SPINK7 mediates EoE pathogenesis primarily through uncontrolled KLK-5 protease activity. Interestingly, the serpin α-1 antitrypsin can inhibit KLK5 activity in vitro and inhibit allergen-induced esophageal eosinophilia in vivo. However, the mechanism of this activity is still unclear and may involve LRP1 instead of or in addition to direct proteolytic inhibition.

[0020] Patients with eosinophilic asthma have low levels of LRP1. Furthermore, deletion of LRP1 specifically in CD11b and CD11c dendritic cells in mice leads to enhanced allergic inflammatory responses in models of allergic airway disease. 29 Mice with a deletion of LRP1 suffered increased antigen uptake, eosinophilic inflammation, allergic sensitization, increased Th2-mediated cytokine production, and reduced T regulatory cells. 29 Therefore, LRP1 may help maintain protease / inhibitor homeostasis in the esophageal environment, mediate TH2 responses, inhibit inflammatory signaling pathways (NFκB, JNK), and result in repair of esophageal dysfunction.

[0021] Serpin peptides and pharmaceutical compositions containing same Disclosed herein are serpin peptides, including isolated synthetic peptides and derivatives thereof, that specifically bind to LRP1. LRP1 is an endocytic scavenger receptor for a number of ligands that exert distinct biological functions. The LRP1 protein consists of a small (85 kD, β-chain) intracellular fragment that spans the cell membrane and is non-covalently linked to an extracellular fragment (515 kD, α-chain) that consists of ligand-binding repeats, which are responsible for the majority of ligand binding. In addition to its ability to mediate endocytosis of various lipoproteins, protease / inhibitor complexes, viruses, matrix proteins and growth factors through its extracellular domain, LRP1 interacts with various scaffolding and signaling proteins through its intracellular domain to mediate cell signaling. Due to its multifunctionality (control of both endocytosis and cell signaling), LRP1 is involved in a variety of biological functions, including cell growth / survival, homeostasis, cell metabolism, cytokine regulation and transport of foreign antigens. Therefore, LRP1 has been implicated in a variety of diseases.

[0022] In some embodiments, the serpin peptide comprises the SP16 peptide (SEQ ID NO: 2). In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 50% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 55% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 60% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 65% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 70% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 75% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 80% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 85% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 90% identity with SP16. In certain embodiments, the serpin peptides disclosed herein are analogs or derivatives of the SP16 peptide that share at least 95% identity with SP16.

[0023] In certain embodiments, the serpin peptides disclosed herein are The serpin peptide comprises, consists essentially of, or consists of the core sequence FNKPFVFLM (SEQ ID NO: 1) of the SP16 peptide having the sequence of TIFF2025503607000005.tif5128. The core sequence comprises an LRP1 binding site having the sequence of FVFLM. Surprisingly, the activity of the serpin peptide was significantly increased when Met of the core sequence was replaced with Nle. Thus, in certain embodiments, the serpin peptide disclosed herein comprises, consists essentially of, or consists of a core binding motif having the sequence X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In certain embodiments, the binding motif is TIFF2025503607000006.tif19164. In certain embodiments, the binding motif has the sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has the sequence of SEQ ID NO: 5. In certain embodiments, the binding motif has the sequence of SEQ ID NO: 6. In certain embodiments, the binding motif has the sequence of SEQ ID NO: 7. In certain embodiments, the binding motif has the sequence of SEQ ID NO: 8. In certain embodiments, the binding motif has the sequence of SEQ ID NO: 9. In certain embodiments, the serpin peptide disclosed herein comprises, consists essentially of, or consists of an LRP1 binding site having the sequence F-X3-X4-X5-X6, where X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In certain embodiments, the LRP1 binding site is In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1. In certain embodiments, the binding motif has a sequence of SEQ ID NO: 1.

[0024] In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of a core binding motif that has been modified by the addition to one or both ends of the core binding motif of flanking sequences that include one or more basic amino acids, arginine, or both basic amino acids and arginine. For example, a serpin peptide disclosed herein comprises, consists essentially of, or consists of an amino acid sequence of Z1-R-X1-N-X2-PF-X3-X4-X5-X6-R-Z2, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, X6 is M, I or Nle, and Z1 and Z2 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 1-3, 1-5, 1-6, 1-7, 1-8, 1-9, or 1-10 basic amino acids. In certain embodiments, Z1 is 1 basic amino acid. In certain embodiments, Z1 is 2 basic amino acids. In certain embodiments, Z1 is 3 basic amino acids. In certain embodiments, Z1 is 4 basic amino acids. In a particular embodiment, Z1 is 5 basic amino acids. In a particular embodiment, Z1 is 6 basic amino acids. In a particular embodiment, Z1 is 7 basic amino acids. In a particular embodiment, Z1 is 8 basic amino acids. In a particular embodiment, Z1 is 9 basic amino acids. In a particular embodiment, Z1 is 10 basic amino acids. In a particular embodiment, Z2 is 1 basic amino acid. In a particular embodiment, Z2 is 2 basic amino acids. In a particular embodiment, Z2 is 3 basic amino acids. In a particular embodiment, Z2 is 4 basic amino acids. In a particular embodiment, Z2 is 5 basic amino acids. In a particular embodiment, Z2 is 6 basic amino acids. In a particular embodiment, Z2 is 7 basic amino acids. In a particular embodiment, Z2 is 8 basic amino acids. In a particular embodiment, Z2 is 9 basic amino acids. In a particular embodiment, Z2 is 10 basic amino acids. In a particular embodiment, Z1 is 1 to 3 basic amino acids.In a particular embodiment, Z1 is 1 to 5 basic amino acids. In a particular embodiment, Z1 is 1 to 6 basic amino acids. In a particular embodiment, Z1 is 1 to 7 basic amino acids. In a particular embodiment, Z1 is 1 to 8 basic amino acids. In a particular embodiment, Z1 is 1 to 9 basic amino acids. In a particular embodiment, Z1 is 1 to 10 basic amino acids. In a particular embodiment, Z2 is 1 to 3 basic amino acids. In a particular embodiment, Z2 is 1 to 5 basic amino acids. In a particular embodiment, Z2 is 1 to 6 basic amino acids. In a particular embodiment, Z2 is 1 to 7 basic amino acids. In a particular embodiment, Z2 is 1 to 8 basic amino acids. In a particular embodiment, Z2 is 1 to 9 basic amino acids. In a particular embodiment, Z2 is 1 to 10 basic amino acids.

[0025] In certain embodiments, the serpin peptides disclosed herein are TIFF2025503607000008.tif19170, where Z1 and Z2 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 1-3, 1-5, 1-6, 1-7, 1-8, 1-9, or 1-10 basic amino acids. In certain embodiments, the serpin peptide comprises, consists essentially of, or consists of the amino acid sequence of TIFF2025503607000009.tif5128. In certain embodiments, the serpin peptide comprises, consists essentially of, or consists of the amino acid sequence of TIFF2025503607000010.tif5128. The present invention comprises, consists essentially of, or consists of the amino acid sequence of TIFF2025503607000011.tif5128. In certain embodiments, Z1 is 1 basic amino acid. In certain embodiments, Z1 is 2 basic amino acids. In certain embodiments, Z1 is 3 basic amino acids. In certain embodiments, Z1 is 4 basic amino acids. In certain embodiments, Z1 is 5 basic amino acids. In certain embodiments, Z1 is 6 basic amino acids. In certain embodiments, Z1 is 7 basic amino acids. In certain embodiments, Z1 is 8 basic amino acids. In certain embodiments, Z1 is 9 basic amino acids. In certain embodiments, Z1 is 10 basic amino acids. In certain embodiments, Z2 is 1 basic amino acid. In certain embodiments, Z2 is 2 basic amino acids. In certain embodiments, Z2 is 3 basic amino acids. In certain embodiments, Z2 is 4 basic amino acids. In a particular embodiment, Z2 is 5 basic amino acids. In a particular embodiment, Z2 is 6 basic amino acids. In a particular embodiment, Z2 is 7 basic amino acids. In a particular embodiment, Z2 is 8 basic amino acids. In a particular embodiment, Z2 is 9 basic amino acids. In a particular embodiment, Z2 is 10 basic amino acids. In a particular embodiment, Z1 is 1 to 3 basic amino acids. In a particular embodiment, Z1 is 1 to 5 basic amino acids. In a particular embodiment, Z1 is 1 to 6 basic amino acids. In a particular embodiment, Z1 is 1 to 7 basic amino acids. In a particular embodiment, Z1 is 1 to 8 basic amino acids. In a particular embodiment, Z1 is 1 to 9 basic amino acids. In a particular embodiment, Z1 is 1 to 10 basic amino acids. In a particular embodiment, Z2 is 1 to 3 basic amino acids. In a particular embodiment, Z2 is 1 to 5 basic amino acids. In a particular embodiment, Z2 is 1 to 6 basic amino acids. In certain embodiments, Z2 is 1 to 7 basic amino acids. In certain embodiments, Z2 is 1 to 8 basic amino acids. In certain embodiments, Z2 is 1 to 9 basic amino acids.In certain embodiments, Z2 is 1 to 10 basic amino acids.

[0026] In certain embodiments, the serpin peptide disclosed herein comprises, consists essentially of, or consists of the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO: 20), wherein X1 is V or L; X2 is R or F; X3 is R or K; X4 is M, Nle or I; Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is any sequence of 5 amino acids.

[0027] In some embodiments, the peptide comprises, consists essentially of, or consists of 20 or fewer amino acids.

[0028] In certain embodiments, the serpin peptide disclosed herein comprises, consists essentially of, or consists of the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO: 21), wherein X1 is V or L; X2 is F or R, X3 is K or R; X4 is V, L or M; X5 is any sequence of 5 amino acids, Z1 is any amino acid, Z2 is any sequence of two amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0029] In some embodiments, the peptide comprises, consists essentially of, or consists of 20 or fewer amino acids.

[0030] In certain embodiments, the serpin peptides disclosed herein are TIFF2025503607000012.tif41165. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 35. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 25. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 29. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 31. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 40. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 41. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 42. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 43. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 44. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 45. In certain embodiments, the serpin peptides disclosed herein comprise, consist essentially of, or consist of the sequence of SEQ ID NO: 46.

[0031] In certain embodiments, the serpin peptides disclosed herein have a size of 5-30 amino acids. For example, the serpin peptides may have a size of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acids. In some embodiments, the serpin peptides may have a size of 5 amino acids. In some embodiments, the serpin peptides may have a size of 6 amino acids. In some embodiments, the serpin peptides may have a size of 7 amino acids. In some embodiments, the serpin peptides may have a size of 8 amino acids. In some embodiments, the serpin peptides may have a size of 9 amino acids. In some embodiments, the serpin peptides may have a size of 10 amino acids. In some embodiments, the serpin peptides may have a size of 11 amino acids. In some embodiments, the serpin peptides may have a size of 12 amino acids. In some embodiments, the serpin peptides may have a size of 13 amino acids. In some embodiments, the serpin peptides may have a size of 14 amino acids. In some embodiments, the serpin peptides may have a size of 15 amino acids. In some embodiments, the serpin peptides may have a size of 16 amino acids. In some embodiments, the serpin peptides may have a size of 17 amino acids. In some embodiments, the serpin peptides may have a size of 18 amino acids. In some embodiments, the serpin peptides may have a size of 19 amino acids. In some embodiments, the serpin peptides may have a size of 20 amino acids. In some embodiments, the serpin peptides may have a size of 21 amino acids. In some embodiments, the serpin peptides may have a size of 22 amino acids. In some embodiments, the serpin peptides may have a size of 23 amino acids. In some embodiments, the serpin peptides may have a size of 24 amino acids. In some embodiments, the serpin peptides may have a size of 25 amino acids.In some embodiments, the serpin peptides may have a size of 26 amino acids. In some embodiments, the serpin peptides may have a size of 27 amino acids. In some embodiments, the serpin peptides may have a size of 28 amino acids. In some embodiments, the serpin peptides may have a size of 29 amino acids. In some embodiments, the serpin peptides may have a size of 30 amino acids. In some embodiments, the serpin peptides disclosed herein have a size of 20 amino acids or less. Longer peptides may decrease solubility, and shorter peptides may decrease stability. As disclosed herein, various modifications can be made to improve stability, such as the addition of poly-R sequences or other flanking sequences to form fusion proteins.

[0032] Serpin peptides include analogs or derivatives thereof. For example, the native sequence of a serpin peptide can be modified to enhance plasma stability and increase the binding affinity of the peptide to its cognate receptor. In certain embodiments, the serpin peptides disclosed herein can be further modified with one or more non-natural peptide bonds or amino acids, or by conjugation to peptide functional groups such as polyethylene glycol (PEG) to extend shelf life and / or bioavailability. In certain embodiments, the serpin peptides disclosed herein are modified by adding one or more amino acid residues, such as arginine, at one or both ends. In certain embodiments, the serpin peptides are modified by adding two, three, or four amino acid residues at both ends. In some embodiments, the serpin peptides are modified by adding two amino acid residues at both ends. In some embodiments, the serpin peptides are modified by adding three amino acid residues at both ends. In some embodiments, the serpin peptides are modified by adding four amino acid residues at both ends.

[0033] In certain embodiments, the serpin peptides disclosed herein may have an N-terminus or C-terminus with additional or modified functional groups. In certain embodiments, one or both of the N-terminus and C-terminus of the serpin peptide may be amidated. In certain embodiments, the C-terminus of the serpin peptide may be amidated. In certain embodiments, the N-terminus of the serpin peptide may be amidated. In certain embodiments, one or both of the N-terminus and C-terminus of the serpin peptide may be acetylated.

[0034] In certain embodiments, the N-terminus of the serpin peptide may be acetylated. For example, in certain embodiments, the serpin peptides disclosed herein are selected from the group consisting of Ac-VKFNKPFVFL(Nle)IEQNTK (N-terminally acetylated SEQ ID NO: 35), Ac-VKFNKPFVFLM (N-terminally acetylated SEQ ID NO: 25), Ac-LRFNRPFLVVI (N-terminally acetylated SEQ ID NO: 29), Ac-VRFNRPFLMII (N-terminally acetylated SEQ ID NO: 31), Ac-VKFNKPFVFL(Nle) (N-terminally acetylated SEQ ID NO: 40), Ac-RFNRPFLVVIR (N-terminally acetylated SEQ ID NO: 41), Ac-RFNRPFLMIIR (N-terminally acetylated SEQ ID NO: 42), Ac-RFNKPFVFL(Nle)R (N-terminally acetylated SEQ ID NO: 43), Ac-RRRFLVVIRRR (N-terminally acetylated SEQ ID NO: Ac-RRRFVFL(Nle)RRR (N-terminal acetylated SEQ ID NO: 44), Ac-RRRFMIIRRR (N-terminal acetylated SEQ ID NO: 45) or Ac-RRRFVFL(Nle)RRR (N-terminal acetylated SEQ ID NO: 46).

[0035] In certain embodiments, the C-terminus of the serpin peptide may be amidated. For example, in certain embodiments, the serpin peptides disclosed herein are selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK-NH2 (C-terminally amidated SEQ ID NO: 35), VKFNKPFVFLM-NH2 (C-terminally amidated SEQ ID NO: 25), LRFNRPFLVVI-NH2 (C-terminally amidated SEQ ID NO: 29), VRFNRPFLMII-NH2 (C-terminally amidated SEQ ID NO: 31), VKFNKPFVFL(Nle)-NH2 (C-terminally amidated SEQ ID NO: 40), RFNRPFLVVIR-NH2 (C-terminally amidated SEQ ID NO: 41), RFNRPFLMIIR-NH2 (C-terminally amidated SEQ ID NO: 42), RFNKPFVFL(Nle)R-NH2 (C-terminally amidated SEQ ID NO: 43), RRRFLVVIRRR-NH2 (C-terminally amidated SEQ ID NO: 44), RRRFLMIIRRR-NH2 (C-terminal amidated SEQ ID NO: 45) or RRRFVFL(Nle)RRR-NH2 (C-terminal amidated SEQ ID NO: 46).

[0036] In certain embodiments, the C-terminus of the serpin peptide may be amidated and the N-terminus acetylated. For example, in certain embodiments, the serpin peptides disclosed herein are Ac-VKFNKPFVFL(Nle)IEQNTK-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 35), Ac-VKFNKPFVFLM-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 25), Ac-LRFNRPFLVVI-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 29), Ac-VRFNRPFLMII-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 31), Ac-VKFNKPFVFL(Nle)-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 40), Ac-RFNRPFLVVIR-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 41), Ac-RFNRPFLMIIR-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: Ac-RFNKPFVFL(Nle)R-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 42), Ac-RFNKPFVFL(Nle)R-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 43), Ac-RRRFLVVIRRR-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 44), RRRFLMIIRRR-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 45) or Ac-RRRFVFL(Nle)RRR-NH2 (C-terminal amidated and N-terminal acetylated SEQ ID NO: 46).

[0037] In certain embodiments, the serpin peptides disclosed herein are fused to one or more other peptides to form a fusion peptide or fusion protein.For example, the one or more other peptides include an epitope tag such as ALFA-tag, V5-tag, Myc-tag, HA-tag, Spot-tag, T7-tag or NE-tag, a half-life extender such as PEG, lipidation, FC fusion or albumin fusion, or both an epitope tag and a half-life extender.In certain embodiments, the peptide includes one or more D-amino acids, i.e., one or more amino acids of the peptide have a D-configuration.

[0038] In another aspect, the present disclosure relates to a pharmaceutical composition comprising, consisting essentially of, or consisting of an effective amount of one or more serpin peptides or fusion peptides disclosed herein. In some embodiments, the pharmaceutical composition further comprises one or more additional therapeutic agents that are not serpin peptides disclosed herein. In some embodiments, the pharmaceutical composition further comprises a pharma- ceutically acceptable carrier, excipient, additive, preservative, or combination thereof. Examples of acceptable carriers include physiologically acceptable solutions, such as sterile saline and sterile buffered saline.

[0039] The term "effective amount" as used herein refers to the amount of a composition that produces a desired effect. An effective amount of a composition can be used to produce a prophylactic or therapeutic effect in a subject, such as preventing or treating a target condition, alleviating symptoms associated with a condition, or producing a desired physiological effect. In such cases, the effective amount of a composition is a "therapeutically effective amount", "therapeutically effective concentration" or "therapeutically effective dose". The correct effective amount or therapeutically effective amount is the amount of the composition that produces the most effective results in terms of efficacy of treatment in a given subject or cell population. This amount will vary depending on a variety of factors, including, but not limited to, the characteristics of the composition (including activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological state of the subject (including age, sex, type and stage of disease, general physical condition, responsiveness to a given dosage, and type of medication) or cell, the nature of the pharmacologic acceptable carrier in the formulation, and the route of administration. Furthermore, the effective amount or therapeutically effective amount can vary depending on whether the composition is administered alone or in combination with another composition, drug, therapy, or other treatment method or modality. Those skilled in the clinical and pharmacological arts will be able to determine an effective or therapeutically effective amount through routine experimentation, i.e., by monitoring the cellular or subject response to administration of the composition and adjusting the dosage accordingly. A "clinically effective amount," "clinically effective concentration," or "clinically effective dose" refers to a concentration or dose of a peptide, composition, or pharmaceutical composition that has been shown to be effective in clinical trials or is predicted to be effective based on early phase or preclinical studies. For further guidance, see Remington: The Science and Practice of Pharmacy, 21 st Edition, Univ. of Sciences in Philadelphia (USIP), Lippincott Williams & Wilkins, Philadelphia, PA, 2005.

[0040] In certain embodiments, the peptide or pharmaceutical composition disclosed herein can be formulated for parenteral administration, such as oral administration, intravenous administration, intramuscular administration, subcutaneous administration (by a device such as a bolus injection or infusion pump), intradermal administration, transdermal administration, topical administration and intranasal administration.In certain embodiments, a subcutaneous infusion pump can be used to deliver the peptide or pharmaceutical composition disclosed herein.The peptide or pharmaceutical composition can be administered more than once.More specifically, after the initial administration, one or more additional doses can be administered as a booster.

[0041] Methods and uses of serpin peptides and pharmaceutical compositions in reducing inflammation, treating pain-related diseases and conditions, and treating diseases associated with A. alternata The serpin peptides or pharmaceutical compositions disclosed herein have various functions. In certain embodiments, disclosed herein is a method of treating a subject in need thereof with an effective amount of one or more serpin peptides, fusion peptides or pharmaceutical compositions disclosed herein. In some embodiments, the subject suffers from a disease or condition related to LRP1 or TSLP. In some embodiments, the subject suffers from acute neuropathic pain, such as acute nociceptive pain, inflammatory pain and neuropathic pain. In some embodiments, the subject suffers from EDD, such as EoE, eosinophilic asthma, atopic dermatitis, nasal polyps and chronic idiopathic urticaria. In some embodiments, the subject suffers from allergic disease, allergic inflammation, or eosinophilic allergic disease.

[0042] As used herein, "treating" a condition or "treatment" of a condition can refer to preventing the condition, slowing the onset or rate of progression of the condition, reducing the risk of developing the condition, preventing or delaying the onset of symptoms associated with the condition, reducing or terminating symptoms associated with the condition, causing complete or partial regression of symptoms, or some combination thereof. Treatment can also refer to prophylactic or preventative treatment of the condition.

[0043] As used herein, the term "subject" refers to a mammal. In some embodiments, the subject is a human. In some embodiments, the subject has not undergone any pretreatment with a serine protease inhibitor, such as alpha-1-antitrypsin treatment, prior to treatment with the peptides disclosed herein.

[0044] In some embodiments, the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions can be administered in humans at a dose of about 0.001 mg / kg to about 4 mg / kg. Depending on the indication, severity and route of administration, an appropriate dose can be appropriately selected. For example, for acute indications, fewer treatments with higher doses are administered at each treatment, while for chronic indications requiring frequent and prolonged treatments, lower doses are administered at each treatment. In some indications, where inflamed tissues express high densities of LRP1, very low doses of serpin peptides such as SP16 and SP163M are required. When subjects suffer from nerve injury, neurons express very high densities of LRP1. In vivo and in vitro studies have demonstrated significant effects at doses as low as 0.05 μg.

[0045] In some embodiments, the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions are administered subcutaneously. In some embodiments, the serpin peptides, such as SP16 or SP163M, are administered subcutaneously to a human subject at a dose of 0.05 mg / kg to 0.5 mg / kg, for example, at a dose of 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.3 mg / kg, 0.35 mg / kg, 0.4 mg / kg, 0.45 mg / kg, or 0.5 mg / kg. In some embodiments, the serpin peptides are administered at a dose of 0.05 mg / kg. In some embodiments, the serpin peptides are administered at a dose of 0.1 mg / kg. In some embodiments, the serpin peptides are administered at a dose of 0.15 mg / kg. In some embodiments, the serpin peptides are administered at a dose of 0.2 mg / kg. In some embodiments, the serpin peptides are administered at a dose of 0.25 mg / kg. In some embodiments, the serpin peptide is administered at a dose of 0.3 mg / kg. In some embodiments, the serpin peptide is administered at a dose of 0.35 mg / kg. In some embodiments, the serpin peptide is administered at a dose of 0.4 mg / kg. In some embodiments, the serpin peptide is administered at a dose of 0.45 mg / kg. In some embodiments, the serpin peptide is administered at a dose of 0.5 mg / kg. In some embodiments, the serpin peptide, such as SP16 or SP163M, is administered subcutaneously to a human subject at a dose of 0.2 mg / kg or 0.4 mg / kg.In some embodiments, the serpin peptide, such as SP16 or SP163M, is orally administered to a human subject at a dose of 1 mg to 150 mg, for example, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, or 150 mg. In some embodiments, the serpin peptide, such as SP16 or SP163M, is orally administered to a human subject at a dose of 5 mg, 25 mg, or 100 mg. In some embodiments, a serpin peptide, such as SP16 or SP163M, is administered locally via a transdermal patch, optionally using pulsatile delivery, e.g., at a dose of 0.05 mg / kg to 0.5 mg / kg, e.g., at a dose of 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.3 mg / kg, 0.35 mg / kg, 0.4 mg / kg, 0.45 mg / kg or 0.5 mg / kg.

[0046] In some embodiments, a single dose of the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions is administered. In some embodiments, the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions are administered as the only therapeutic agent. In some embodiments, the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions are administered in combination with a second therapeutic agent.

[0047] In some embodiments, the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions are administered in a pulsatile or continuous mode, hi some embodiments, the serpin peptides and fusions thereof or pharmaceutical compositions comprising said peptides or fusions are administered via a transdermal patch, an inhaler, or an intranasal device.

[0048] Methods of reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP by administering a serpin peptide to a subject in need thereof In some embodiments, the present technology includes a method of reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP. In some aspects, the method includes administering to a subject in need thereof any of the serpin peptides described in the present disclosure.

[0049] In some aspects, the serpin peptide administered to reduce inflammation in a subject having an LRP1 or TSLP associated disease or condition is TIFF2025503607000013.tif48165.

[0050] In some aspects, the serpin peptide administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises SP16 peptide (SEQ ID NO: 2) or SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 35).

[0051] In some aspects, the serpin peptide administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises the amino acid sequence of X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. Contains the sequence TIFF2025503607000014.tif19164.

[0052] In some aspects, the serpin peptide administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO:20), where X1 is V or L, X2 is R or F, X3 is R or K, X4 is M, Nle or I, Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is a sequence of any 5 amino acids. In some aspects, the serpin peptide comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO:21), where X1 is V or L, X2 is F or R, X3 is K or R, X4 is V, L or M, X5 is a sequence of any 5 amino acids, Z1 is any amino acid, Z2 is a sequence of any 2 amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0053] In some aspects, the serpin peptide administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises the sequence of SEQ ID NO: 35. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 2. In some aspects, the N-terminus of the serpin peptide is acetylated. In some aspects, the C-terminus of the serpin peptide is amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In some aspects, the one or more other peptides are different from the serpin peptide. In some aspects, the fusion peptide or fusion protein comprises the serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the technology includes a pharmaceutical composition comprising a serpin peptide and a pharma- ceutically effective carrier.

[0054] In some embodiments, the serpin peptide administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP is administered at a therapeutically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a therapeutically effective dose. In some aspects, the serpin peptide is administered at a clinically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a clinically effective dose. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human. In some aspects, administration is by oral, parenteral, intradermal, transdermal, topical, or intranasal administration. In some aspects, the composition is administered as a single dose. In some aspects, the serpin peptide is administered by topical administration. In some aspects, the serpin peptide is administered by oral administration.

[0055] In some aspects, the serpin peptide is administered to reduce inflammation in a subject with a disease or condition associated with LRP1 or TSLP, the disease or condition being caused by A. alternata. In some aspects, the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia. In some aspects, the disease or condition is rhinitis. In some aspects, the disease or condition is asthma. In some aspects, the disease or condition is dermatitis. In some aspects, the disease or condition is esophageal eosinophilia. In some aspects, the disease or condition is acute or neuropathic pain. In some aspects, the disease or condition is acute nociceptive, inflammatory, or neuropathic pain. In some aspects, the disease or condition is EDD. In some aspects, the disease or condition is EoE, eosinophilic asthma, atopic dermatitis, nasal polyps, or chronic idiopathic urticaria. In some aspects, the disease or condition is atopic dermatitis. In certain embodiments, the disease or condition is pruritus.In some aspects, the disease or condition is an allergic reaction.In some aspects, the disease or condition is allergic inflammation.In some aspects, the disease or condition is an eosinophil-induced allergic disease.In some aspects, the disease or condition is caused by TH2-type inflammatory cytokines.

[0056] Methods for treating acute or neuropathic, nociceptive, or inflammatory pain by administering a serpin peptide to a subject in need thereof In some embodiments, the technology includes a method of treating a subject with a disease or condition associated with LRP1 or TSLP, which is acute or neuropathic pain, nociceptive pain, or inflammatory pain. In some aspects, the technology includes a method of treating a subject with a disease or condition associated with LRP1, which is acute or neuropathic pain, nociceptive pain, or inflammatory pain. In some aspects, the method includes administering to a subject in need thereof any of the serpin peptides described in the present disclosure.

[0057] In some aspects, a method for treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises: The method includes administering a serpin peptide comprising an amino acid sequence selected from the group consisting of: TIFF2025503607000015.tif48165. In some aspects, the serpin peptide is administered to the subject to treat a disease or condition associated with LRP1 or TSLP. In some aspects, the serpin peptide is administered to the subject to treat a disease or condition associated with LRP1.

[0058] In some aspects, a method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a serpin peptide comprising SP16 peptide (SEQ ID NO: 2) or SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 35).

[0059] In some aspects, a method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a serpin peptide comprising the amino acid sequence of X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In some aspects, the serpin peptide is Contains the sequence TIFF2025503607000016.tif19164.

[0060] In some aspects, a method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a serpin peptide comprising the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO: 20), where X1 is V or L, X2 is R or F, X3 is R or K, X4 is M, Nle or I, Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is a sequence of any 5 amino acids. In some aspects, the serpin peptide comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO:21), where X1 is V or L, X2 is F or R, X3 is K or R, X4 is V, L or M, X5 is a sequence of any 5 amino acids, Z1 is any amino acid, Z2 is a sequence of any 2 amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0061] In some aspects, a method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering a serpin peptide comprising the sequence of SEQ ID NO: 35. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 2. In some aspects, the N-terminus of the serpin peptide is acetylated. In some aspects, the C-terminus of the serpin peptide is amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In some aspects, the one or more other peptides are different from the serpin peptide. In some aspects, the fusion peptide or fusion protein includes the serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the technology includes a pharmaceutical composition including a serpin peptide and a pharma- ceutically effective carrier.

[0062] In some aspects, the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a serpin peptide at a therapeutically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a therapeutically effective dose. In some aspects, the serpin peptide is administered at a clinically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a clinically effective dose. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human. In some aspects, administration is by oral, parenteral, intradermal, transdermal, topical, or intranasal administration. In some aspects, the composition is administered as a single dose. In some aspects, the serpin peptide is administered by topical administration. In some aspects, the serpin peptide is administered by oral administration.

[0063] In some aspects, the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a serpin peptide to treat a disease or condition associated with LRP1. In some aspects, the disease or condition is acute or neuropathic pain. In some aspects, the disease or condition is nociceptive pain. In some aspects, the disease or condition is inflammatory pain. In some aspects, administering the serpin peptide results in the alleviation of pain. In some aspects, administering the serpin peptide prevents or reduces the onset of pain. In some aspects, administering the serpin peptide results in an increase in neuronal survival and neurite sprouting.

[0064] Methods for treating a disease or condition caused by A. alternata by administering a serpin peptide to a subject in need thereof In some embodiments, the technology includes a method of treating a subject having a disease or condition associated with LRP1 or TSLP, wherein the disease or condition is caused by A. alternata. In some aspects, the technology includes a method of treating a subject having a disease or condition associated with TSLP, wherein the disease or condition is caused by A. alternata. In some aspects, the method includes administering any of the serpin peptides described in the present disclosure to a subject having a disease or condition caused by A. alternata.

[0065] In certain embodiments, the method of treating a disease or condition caused by A. alternata comprises administering to a subject, to treat the disease or condition caused by A. alternata, The method includes administering a serpin peptide comprising an amino acid sequence selected from the group consisting of: TIFF2025503607000017.tif48164.

[0066] In some aspects, a method of treating a disease or condition caused by A. alternata comprises administering a serpin peptide comprising SP16 peptide (SEQ ID NO: 2) or SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with the SP163M peptide (SEQ ID NO: 35).

[0067] In some aspects, a method of treating a disease or condition caused by A. alternata comprises administering a serpin peptide comprising the amino acid sequence of X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In some aspects, the serpin peptide is Contains sequence TIFF2025503607000018.tif19164.

[0068] In some aspects, a method of treating a disease or condition caused by A. alternata comprises administering a serpin peptide comprising the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO: 20), where X1 is V or L, X2 is R or F, X3 is R or K, X4 is M, Nle or I, Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is a sequence of any 5 amino acids. In some aspects, the serpin peptide comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO:21), where X1 is V or L, X2 is F or R, X3 is K or R, X4 is V, L or M, X5 is a sequence of any 5 amino acids, Z1 is any amino acid, Z2 is a sequence of any 2 amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0069] In certain embodiments, a method of treating a disease or condition caused by A. alternata comprises administering a serpin peptide comprising the amino acid sequence of SEQ ID NO: 35. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 2. In certain embodiments, the N-terminus of the serpin peptide is acetylated. In certain embodiments, the C-terminus of the serpin peptide is amidated. In certain embodiments, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In certain embodiments, the one or more other peptides are different from the serpin peptide. In certain embodiments, the fusion peptide or fusion protein comprises the serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the technology comprises a pharmaceutical composition comprising a serpin peptide and a pharma- ceutically effective carrier.

[0070] In some aspects, a method of treating a disease or condition caused by A. alternata includes administering a serpin peptide at a therapeutically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a therapeutically effective dose. In some aspects, the serpin peptide is administered at a clinically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a clinically effective dose. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human. In some aspects, administration is by oral, parenteral, intradermal, transdermal, topical, or intranasal administration. In some aspects, the serpin peptide is administered as a single dose.

[0071] In some aspects, a method of treating a disease or condition caused by A. alternata comprises administering a serpin peptide, wherein the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia. In some aspects, the disease or condition is rhinitis. In some aspects, the disease or condition is asthma. In some aspects, the disease or condition is dermatitis. In certain embodiments, the disease or condition is esophageal eosinophilia. In some aspects, administering the serpin peptide reduces inflammation. In some aspects, administering the serpin peptide reduces eosinophilic inflammation. EXAMPLES

[0072] The following examples are intended to illustrate various aspects of the present invention.Therefore, the specific aspects discussed should not be interpreted as limitations on the scope of the present invention.It is clear to those skilled in the art that various equivalents, changes and modifications can be made without departing from the scope of the present invention, and it is understood that such equivalent aspects are included in this specification.In addition, all references cited in this disclosure are incorporated herein by reference in their entirety as if fully set forth herein.

[0073] material and method animal: Male Sprague Dawley rats (170–200 g; 8–12 weeks old) and C57BL / 6J mice (25 g; male and female, 8–12 weeks old) were purchased from Envigo and Jackson Laboratory, respectively. All animal experiments were approved by the University of California, San Diego, Animal Care and Use Committee. All rats and mice were housed under a 12:12 h light:dark cycle and had free access to food and water.

[0074] reagent: TIFF2025503607000019.tif5128 was kindly provided by Serpin Pharma (Manassas, VA, USA). Briefly, peptides were synthesized by CPC Scientific Inc (Sunnyvale, CA) and purity was confirmed to be >95% by high performance liquid chromatography and mass spectrometry. Recombinant human EI-tPA was purchased from Molecular Innovations (Novi, MI, USA). NGF-β was purchased from Sigma (St. Louis, MO, USA). Receptor-associated proteins were expressed as glutathione-S-transferase (GST) fusion proteins (GST-RAP).

[0075] Neurite outgrowth in primary cultures of adult DRG neurons: Primary DRG neurons were isolated from adult male Sprague Dawley rats and cultured as previously described for mice with modifications. 53DRGs were detached from the nerve roots and collected in Hank's Buffered Salt Solution (HBSS) on ice. DRGs were enzymatically digested and approximately 4000 DRG neurons were plated in each well of a 12-well tissue culture plate (Thermo Fisher Scientific, Waltham, MA, USA). All DRG neurons were cultured in DMEM / F12 containing 2% B27 and 1% FBS for 54 h at 37°C in 5% CO2, and vehicle or SP163M (0–500 ng / ml) was added every 24 h. Primary cultured DRG neurons were imaged by phase contrast, and cell viability was assessed by trypan blue. Primary DRG neurons were cultured and fixed in 4% paraformaldehyde and immunofluorescence was performed using mouse anti-βIII-tubulin primary antibody (Promega, Madison, WI, USA; Cat. No. G7121, 1:250) followed by Alexa Fluor-488 anti-mouse antibody (Life Technologies, Carlsbad, CA, USA) as secondary antibody. DRG neurons were manually imaged at 20x and 40x and the longest neurite length per cell was measured in 11 images from multiple wells and separate experiments. Approximately 222 and 144 neurons were measured in the SP16 and control groups, respectively. Quantification was performed in a blinded manner. All neurite outgrowth measurements were performed in duplicate with at least six separate experiments.

[0076] Cell Signaling Analysis: Rat PC12 cells were purchased from ATCC (CRL-1721). PC12 cells were maintained in high glucose DMEM (Gibco, USA) containing 10% heat-inactivated FBS (Gibco, USA), 5% HyClone heat-inactivated horse serum (Cytiva, USA), penicillin (100 units / ml) and streptomycin (1 mg / ml) in 6-well plates pre-coated with 0.01 mg / ml type IV collagen (Sigma-Aldrich, St. Louis, MO, USA). Cells were transferred to serum-free medium (SFM) 4 h prior to effector addition and then treated with SP163M (2.4, 24 or 240 nM), EI-tPA (12 nM), NGF (0.36 nM), or vehicle (PBS) for 10 min. In some cases, cells were preincubated with GST-RAP (150 nM), a competitive antagonist of LRP1, for 15 min or electroporated using the Rat Neuron Nucleofector Amaxa Kit (Lonza Biosciences) and incubated for 48 h with siRNA to silence LRP1 expression (siLRP1; M-094191-01-0010, Dharmacon). Control cells were transfected with non-targeting control (NTC) siRNA (NTC; D-001810-10-05, Dharmacon). Cells were rinsed in ice-cold PBS and proteins were extracted in RIPA buffer (20 mM sodium phosphate, 150 mM NaCl, pH 7.4, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS) supplemented with protease and phosphatase inhibitors (Roche Diagnostics, USA). After 30 min on ice, the lysates were centrifuged at 15000 × g for 5 min, and the supernatants were collected and stored at −20° C. Equal amounts of protein (20 μg) from cell lysates, as determined by BCA protein assay (ThermoFisher Scientific, Waltham, MA, USA), were subjected to 10% SDS-PAGE and electrophoretically transferred to nitrocellulose membranes.Membranes were blocked with 5% nonfat dry milk and then incubated with primary anti-phospho-AKT (Cell Signaling Technology, Danvers, MA, USA; Catalog No. 9271S; 1:1000), anti-phospho-ERK1 / 2 (Cell Signaling Technology, Danvers, MA, USA; Catalog No. 9101S; 1:1000), anti-LRP1 (Cell Signaling Technology, Danvers, MA, USA; Catalog No. 64099S; 1:1000) or anti-total-ERK1 / 2 (Cell Signaling Technology, Danvers, MA, USA; Catalog No. 91012S; 1:1000). Immunoblots were developed with Radiance, Radiance Q and Radiance Plus chemiluminescent substrates and imaged using a BioRad ChemiDoc Imaging System (Bio-Rad, Hercules, CA, USA).

[0077] RT-qPCR: RNA was isolated from DRG cultures using the NucleoSpin RNA kit (MachereyNagel, Duren, GER) and reverse transcribed using the iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA). qPCR was performed using TaqMan Gene Expression Products (ThermoFisher Scientific, Waltham, MA, USA) for GAP-43 (Rn01474579), LRP1 (Rn01503901_m1) and GAPDH (Rn99999916_s1). Amplification was performed with a CFX Connect Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA). Relative changes in mRNA expression were calculated using a 2:1 ratio with GAPDH mRNA as an internal normalization factor, as previously described. ΔΔCT The calculation was performed using the method 56 .

[0078] Intraplantar formalin and capsaicin model: Male mice (n = 33) were acclimated to the behavioral testing facility for at least 60 min. Mice were randomly divided into four groups and administered SP163M (0.02, 0.2 and 2 μg / g) or vehicle subcutaneously. One hour later, 20 μl of 2.5% formalin was injected subcutaneously into the plantar area of ​​the left hind paw. Immediately after formalin injection, mice were placed in a Plexiglas box (22 × 22 × 14 cm). Two observers blinded to treatment recorded the total time mice spent licking and responding to the left hind paw every 5 min over a 1-h period. To quantify the formalin response, activity during the early phase (0–10 min) and late phase (15–50 min) were examined separately.

[0079] For the capsaicin test, male (n = 41) and female (n = 37) mice were acclimated to the behavioral testing facility for at least 1 h. Capsaicin was dissolved in 20% (2-hydroxypropyl)-β-cyclodextrin (Sigma-Aldrich, St. Louis, MO, USA) solution. This vehicle concentration solubilized capsaicin and did not induce a behavioral response when administered alone. Vehicle, SP163M (2 μg / g; sc) or enzymatically inactive tPA (EI-tPA; 2 μg / g iv) were administered 1 h prior to intraplantar injection. Then, 10 μl of 2 μg / μl capsaicin solution was injected into the plantar left hind paw. Immediately after capsaicin injection, mice were placed in a Plexiglas box. Two observers blinded to treatment recorded the time spent licking and responding to the left hind paw over a 10-min period.

[0080] Neuropathic Pain Models: Mice (n = 30) were randomly assigned to two different groups: SP163M (2 μg / g; sc 100 μl) and vehicle (H20 sc 100 μl). Mice were treated 1 h before partial nerve ligation (PNL) and then daily for 2 weeks at least 1 h before behavioral testing. PNL testing was performed as previously described. 59, adapted for mouse 57 Male mice were anesthetized with 3% isoflurane (Vetone, USA) in 1.5 L / min oxygen (Praxair, USA) and maintained with 2.5% isoflurane. An incision was made along the long axis of the femur. The sciatic nerve was exposed at the mid-thigh by dissecting the biceps femoris and superficial gluteus muscles and then carefully removing the surrounding connective tissue. A 9-0 nylon suture (Ethicon, Inc., Somerville, NJ, USA) was inserted into the nerve and ligated to include 1 / 3–1 / 2 of the nerve. The muscle and skin layers were closed using Reflex7 7 mm stainless steel wound clips (CellPoint Scientific, Inc., Gaithersburg, MD, USA). For behavioral testing, mice were acclimated and baseline testing was performed 1 week prior to PNL. Mechanical sensitivity (tactile allodynia) was tested by applying 0.04–4 g Von Frey filaments (Stoelting, Wood Dale IL, USA) to the plantar surface of the ipsilateral hindpaw. The filaments were used as previously described. 60 , presented sequentially in either ascending or descending order using an up-down method modified for mice. 57,61 The filament that elicited paw withdrawal with a 50% probability (50% PWT) was determined. Tactile allodynia was tested on days 2, 4, 9, 11 and 14 after PNL. Results were averaged and subjected to statistical analysis. All experiments were performed by an investigator blinded to the identity of the mice.

[0081] Immunoblotting of sciatic nerve: Sciatic nerves were harvested 2 days after PNL to identify early molecular and cellular changes. Approximately 0.5 cm of sciatic nerve was collected distal to the ligation site. Ipsilateral and contralateral nerves were collected. Nerves were lysed in RIPA buffer, and equal amounts of protein (20 μg) from cell lysates, as determined by BCA protein assay (Bio-Rad, Hercules, CA, USA), were subjected to 10% SDS-PAGE and electrophoretically transferred to nitrocellulose membranes. Membranes were blocked with 5% nonfat dry milk and then incubated with anti-TLR4(CD284) / MD2 (BioLegend, San Diego, CA, USA; Cat. No. 117601, 1:1000), anti-CD11b (Abcam, Cambridge, MA, USA, Cat. No. Ab1333357) and anti-β-actin (Cell Signaling Technology, Danvers, MA, USA; Cat. No. 1:1000). Primary antibodies were detected with HRP-conjugated species-specific secondary antibodies (Cell Signaling Technology, Danvers, MA, USA; Cat. No. 7076S or 4S; 1:5000). Immunoblots were developed using SuperSignal West Pico PLUS chemiluminescent substrate (Thermo Fisher Scientific, Waltham, MA, USA) and a Protec Ecomax X-ray film processor. Densitometric analysis was performed using Image J software (US National Institutes of Health, Bethesda, MD, USA).

[0082] DRG immunohistochemistry: DRGs were embedded in paraffin. For IHC studies, 4-μm-thick DRG tissue sections were immunostained for CD11b (Abcam, Cambridge, MA, USA, Catalog No. Ab1333357; 1:4500) or GFAP (Dako, Santa Clara, CA, USA; Catalog No. $Z0334; 1:4000). Slides were immunostained using Ventana Discovery Ultra (Ventana Medical Systems, Oro, AZ, USA). Antigen retrieval was performed with CC1 (Tris-based; pH 8.5) at 95°C for 40 min. Primary antibodies CD11b and GFAP were incubated with slides at 37°C for 32 min. Secondary antibody OmniMap anti-HRP (Ventana Medical Systems, Oro, AZ, USA; Catalog No. 760-4311) was incubated on sections at 37°C for 12 min. Antibodies were visualized using diaminobenzidine as a chromogen followed by hematoxylin as a counterstain. Slides were rinsed, dehydrated in alcohol and xylene, and coverslipped. Light microscopy was performed using a Leica DFC420 microscope equipped with Leica Imaging Software 2.8.1 (Leica Biosystems, Vista, CA, USA).

[0083] Statistical analysis: Statistical analysis was performed using GraphPad Prism (GraphPad Prism version 9.1.2 for Mac, GraphPad Software, San Diego, CA, USA). All results were expressed as mean ± SEM. Comparisons between two groups were performed using two-tailed unpaired T-tests. Nonparametric Mann-Whitney U-tests were used when the variances of the two populations were significantly different. When comparing more than two groups, one-way ANOVA with Tukey's post-hoc test was performed or, for nonparametric data, Kruskal-Wallis test was utilized. For measurements of neuropathic pain, multiple observations were made over time in individual mice and were collected and analyzed by repeated measures ANOVA with Sidak's post-hoc test. p < 0.05 was considered statistically significant.

[0084] Example 1: Identification of anti-inflammatory motifs As demonstrated herein, a small peptide fragment from the C-terminus of α-1 antitrypsin (the prototypic serpin) can bind to LRP1 and exert potent cell regenerative, tissue protective and immunomodulatory functions. Interestingly, a naturally occurring C-terminal degradation product of α-1 antitrypsin (designated C-36) exhibits proinflammatory activity similar to lipopolysaccharide in both macrophages and neutrophils. 44 Anti-inflammatory sequences were identified by truncating short fragments from the C-terminus of α-1 antitrypsin. The amino acid sequences of the peptides tested are shown in Table 1 below. The core sequence containing the LRP1 binding site of each peptide is shown in bold and underlined.

[0085] Table 1. Peptides tested for anti-inflammatory activity TIFF2025503607000020.tif142170

[0086] Reporter cells (THP1-XBlue-MD2-CD14 cells) were treated with each peptide (50 μg / ml) and then stimulated with LPS (5 ng / ml) and incubated overnight. NFkB-inducible secreted embryonic alkaline phosphatase (SEAP) was measured in the supernatant and absorbance was read. As shown in Figure 1, various serpin peptides sharing a common core motif exhibited anti-inflammatory activity in NFkB reporter cells upon stimulation with LPS. LPS stimulation led to an increase in NFkB activity, and neither the scrambled core control peptide nor the AAT C-36 fragment reduced NFkB activity. Indeed, the AAT C-36 fragment exhibited NFkB-inducing properties without the need for LPS.

[0087] In contrast, SP16 was able to reduce NFkB activity, whereas the short core peptide SP20 had no inhibitory effect. This truncated AAT-derived peptide, which contains the core sequence and the LRP1 binding site, was unstable and thus inactive. Flanking SP16 on both sides with three arginine amino acids (termed "polyR") to obtain SP21 increased the stability of the SP21 peptide and increased NFkB inhibition compared to SP16. Stabilization of the short core of SP16, the SP20 peptide, with three arginine flanking segments to obtain SP22 significantly reduced NFkB activity. Similar effects were observed with two other pairs of serpin peptides, serpin2 short core (SP24) vs. serpin2 short core polyR (SP26) and serpin3 short core (SP28) vs. serpin3 short core polyR (SP29). Thus, this example shows that shortening the peptide resulted in instability and loss of function, whereas stabilization with polyR flanking segments enhanced activity.

[0088] Furthermore, all of these peptides contain the LRP1-binding site, but truncation of the LRP1-binding site to obtain SP31 abolished the anti-inflammatory activity of this peptide and it could not be restored by the polyarginine-flank (SP32 peptide), suggesting that the serpins contain an LRP1-dependent anti-inflammatory core motif.

[0089] Example 2: Primary adult sensory neurons sprout in response to SP163M To determine whether SP163M possesses bioactivity in sensory neurons, primary adult rat DRG neurons were treated with SP163M for up to 96 hours. Within 48 hours, phase contrast images of DRG neuron cultures revealed that SP163M induced higher neuronal survival and neurite sprouting compared to untreated controls (Figure 2A). After 72 and 96 hours in culture, neuronal networks became extensive and continued to show higher neuronal soma survival and neurite outgrowth.

[0090] To specifically identify neurons, immunofluorescence studies with βIII-tubulin were performed. Primary adult DRG neurons cultured on poly-L-lysine (PLL) and laminin showed basal levels of sprouting when treated with vehicle after 54 hours (Figure 2B). In contrast, cultured DRG neurons treated with SP163M showed significant levels of sprouting, including an increase in both branching and length, after 54 hours. Quantification of the longest neurites suggested that SP163M was significantly neurotrophic (Figure 2C). RT-qPCR analysis of the growth-associated protein GAP-43, a regeneration-associated gene, demonstrated that SP16 increased GAP-43 mRNA compared to vehicle-treated neurons (Figure 2D).

[0091] Example 3: SP163M activates LRP1-dependent cell signaling Previously, LRP1 ligands such as EI-tPA have been shown to potently activate cell survival signaling in neurons and SCs, but the effect of SP163M was unknown. First, several concentrations of SP163M were added to cultured PC12 cells for 10 min. SP16 activated phospho-ERK1 / 2 at concentrations as low as 24 nM (Fig. 3A). Next, SP16 was added to PC12 cells over time. SP16 potently activated ERK1 / 2 from 5 to 30 min (Fig. 3B). The activation of ERK1 / 2 by SP16 at 10 min was similar to that of EI-tPA, a known LRP1 interactor. EI-tPA is a derivative of the protease tissue-type plasminogen activator (tPA), an activator of fibrinolysis, and is approved worldwide for the treatment of nonhemorrhagic stroke. Next, LRP1 expression was silenced with siRNA (siLRP1). PC12 cells transfected with siLRP1 showed significantly decreased (70%) LRP1 mRNA expression compared to cells transfected with NTC siRNA for 48 h (Figure 3C). LRP1 protein was also significantly reduced after 48 h (Figure 3D). Treatment of NTC siRNA-transfected PC12 cells with SP16 potently activated ERK1 / 2. EI-tPA also potently activated ERK1 / 2 as expected. In contrast, ERK1 / 2 was not activated by SP163M or EI-tPA in cells transfected with LRP1-specific siRNA (Figure 3E). SP163M also activated Akt and EKR1 / 2 after 10 min in PC12 cells, but this was blocked by the addition of the 39 kDa LRP1 antagonist RAP (150 nM), as expected for any LRP1-dependent agonist (Figure 3F). NGF also activated ERK1 / 2 as expected, serving as a cell signaling control. Collectively, these data support that SP163M is bioactive in sensory and sensory-like neurons through an LRP1-dependent pathway.

[0092] Example 4: SP163M modulates primary afferent input and reduces central sensitization in the late phase of the formalin test The formalin test is a tissue injury model with an acute nociceptive first phase and an inflammatory second phase. 62 It is a widely used tool for screening analgesic and anti-inflammatory pain therapeutics. We tested whether LRP1 agonism by SP163M modulates pain responses induced by intraplantar formalin (Figure 4A). SP163M (0.02, 0.2 or 2.0 μg / g) or vehicle was administered 1 h prior to paw injection of 2.5% formalin solution. Preemptive administration was first studied to understand the contribution of primary afferent inputs in the early phase, an approach consistent with other formalin test studies testing the early effects of novel opioids or other analgesics. 62,63 Time spent licking was quantified over a 50-min period by two blinded observers. Vehicle-treated mice exhibited a characteristic paw-licking pattern during two phases. During the early phase (0–10 min), vehicle-treated mice licked their injected hind paw vigorously for over 100 s. However, SP163M-treated mice showed a dose-dependent reduction (0.02 μg / g; 80 s) and increase (2 μg / g; 60 s) in licking time. Analysis of the area under the curve (AUC) revealed that both the low and high SP163M-treated groups reached statistical significance (Figure 4B). During the later phase (15–50 min), vehicle-treated mice showed, as expected, paw licking that peaked after 25 min (90 s) and disappeared by 50 min (Figure 4A). In contrast, the highest dose of SP163M (2 μg / g) delayed the peak onset time of the late phase to 30 min. Analysis of the AUC revealed that both the medium dose (0.2 μg / g) and the high dose (2 μg / g) reduced licking time in the late phase compared to the vehicle control (Figure 4C). These findings suggest that activation of LRP1 by SP16 reduced both the nociceptive and ongoing sensitization / inflammatory phases of the formalin test.

[0093] Example 5: LRP1 ligands block acute nociception Capsaicin, a lipophilic vanilloid compound found in "hot" chili peppers64 When injected intraplantarly, it binds to and activates transient receptor potential vanilloid 1 (TRPV1) in nociceptive peripheral terminals. 65 This induces ion influx and action potential firing associated with burning pain, resulting in a licking response of the injected paw. Acute spontaneous pain-related behavior induced by capsaicin is transient (less than 10 min), with most of the activity occurring during the first 2-3 min. Initially, it was confirmed that capsaicin increases pain-related behavior in male and female mice when 20% cyclodextrin was used as a vehicle for the capsaicin solution. Both male and female mice showed a greater licking response to capsaicin injection than to vehicle injection, although males tended to be slightly more sensitive (Figure 5A).

[0094] As antinociceptive activity has not been previously shown for any LRP1 interactor, both EI-tPA and SP163M were tested. EI-tPA or SP16 was delivered systemically and anticipatory 1 h before capsaicin injection in both male and female mice. In male mice, SP163M and EI-tPA blocked capsaicin-induced acute pain-related behaviors (Figure 5B). Similar effects of LRP1 agonists were observed in female mice (Figure 5C).

[0095] Example 6: SP163M inhibits the development of neuropathic pain, suppresses innate immunity, and reduces recruitment of inflammatory cells SP163M was then tested in a neuropathic pain model, PNL. PNL of the sciatic nerve induces mechanical hypersensitivity that can be observed within 2 days after injury. 57Male mice were randomly divided into groups and baselined with von Frey filaments and paw withdrawal thresholds (PWT) were recorded 1 week prior to PNL. SP163M (2 μg / g) was administered 1 h prior to PNL and daily for 2 weeks to test whether SP163M was neuroprotective. As expected, vehicle-treated mice developed tactile allodynia by day 2, and hypersensitivity persisted for 2 weeks (Figure 6A). In contrast, mice treated with SP163M did not develop mechanical hypersensitivity. Statistically, the effect of SP163M was most pronounced early after injury (days 2 and 4), although the antiallodynic effect of SP163M persisted until day 9 after injury. These results suggest that activation of LRP1 can abrogate the development of mechanical sensitivity induced by direct nerve injury.

[0096] It has been reported that LRP1 can induce potent anti-inflammatory activity in macrophages during myocardial infarction and modulate innate immunity. To determine whether SP163M modulates neuroinflammation in the injured PNS, sciatic nerves were harvested from vehicle- and SP163M-treated groups 2 days after PNL. CD11b was used to identify inflammatory cells present within the nerve. As expected, CD11b was robustly increased in the nerve immediately distal to the ligation site in vehicle-treated mice (Figure 6B). In contrast, CD11b was decreased in SP163M-treated nerves. Densitometric analysis revealed that CD11b was increased nearly 20-fold (Figure 6C, p < 0.001) and SP163M reduced the levels of CD11b by 10-fold (p < 0.01). Because TLR4 activation is associated with pain states, we also measured TLR4 in the injured sciatic nerve. Immunoblots revealed that TLR4 was upregulated in nerves 2 days after PNL and that SP16 potently downregulated TLR4 expression (Fig. 6D). Densitometric analysis revealed that TLR4 increased more than two-fold after injury in vehicle-treated mice and that SP163M completely blocked TLR4 upregulation (Fig. 6E).

[0097] Recently, it has been shown that inflammatory cells and macrophages that acutely infiltrate the DRG after nerve injury directly regulate chronic pain states. Therefore, L4 DRGs were harvested from mice that received vehicle or SP163M treatment 2 days after PNL. To identify CD11b, immunohistochemistry was performed on transverse sections of DRG (Figure 7A). In PNL sections, CD11b immunoreactivity was observed between neuronal cell bodies and around blood vessels.

[0098] In contrast, transverse sections of nerves treated with SP163M revealed little immunoreactivity, indicating the presence of very low levels of inflammatory cells. Quantification of CD11B immunohistochemistry showed that SP163M-treated DRGs had approximately 5-fold lower CD11b levels than vehicle-treated DRGs (Figure 7B). We next examined satellite cell activation. Satellite cells in naive DRGs do not express GFAP, but after injury, satellite cells express GFAP abundantly. Two days after PNL, satellite cells showed strong GFAP immunoreactivity in vehicle-treated DRGs. Conversely, mice treated with SP163M showed significantly lower GFAP expression in DRGs. Quantification of GFAP immunohistochemistry revealed a 6-fold decrease in GFAP levels in mice treated with SP163M (Figure 7C). These findings suggest that LRP1 engagement reduces satellite cell activation.

[0099] Thus, Examples 2-6 demonstrate the potent efficacy of SP163M in three different preclinical mouse models including acute nociceptive pain, inflammatory pain, and neuropathic pain. Central to the preventive effect of SP163M on mechanical hypersensitivity was its potent anti-inflammatory activity in injured peripheral nerves. In these studies, SP163M potently reduced early recruitment of inflammatory cells distal to the nerve injury site and in the corresponding L3, L4 DRG early after sciatic nerve ligation. SP163M may delay and / or limit the infiltration of inflammatory cells, thereby modulating the pain state.

[0100] Example 7: SP163M blocks IL-13-stimulated Stat6 phosphorylation Esophageal EPC2 cells were treated with vehicle (ddw), A1AT, or SP163M. Cells were harvested at the indicated time points and Western blot analysis of phosphorylated specific STAT6 was performed. Thirty minutes after IL-13 induction, STAT6 was phosphorylated in vehicle and A1AT-treated cells, but not in SP163M-treated cells (Figure 8A). This reduction in STAT6 phosphorylation by SP163M was sustained for several hours after treatment (many different experimental replicates shown) (Figure 8B). Figure 8C shows that the reduction in phosphorylated STAT6 by SP163M was dependent on the expression of LRP1. Using CRISPER / CAS9 technology, LRP1 knockout esophageal EPC2 cell lines were generated. In control cells (with LRP1), SP163M was able to reduce phospho-STAT6 expression, but in LRP1 knockout cell lines, SP163M was unable to reduce phosphorylated STAT6.

[0101] Example 8: SP163M exhibits inhibitory effects in an additional eosinophilic esophagitis model using A. alternata as an allergen Balb / C mice were repeatedly challenged with a series of allergens, A. alternata, over a 4-week period. A. alternata is a common airborne mold associated with eosinophilic inflammation and various allergic diseases, such as rhinitis, asthma, and dermatitis, and is an established model for inducing allergic responses in the esophagus. In this model, allergen challenge leads to increased esophageal eosinophilia. SP163M, A1AT, or vehicle (control) were administered twice weekly for 4 weeks for a total of eight treatments. At the end of the study, esophageal sections were stained to detect MBP (eosinophil marker) and quantified per high-power field. Treatment with SP163M reduced the number of eosinophils infiltrating the esophagus compared to both control mice treated with vehicle as a control and mice treated with a higher dose of A1AT than SP163M (Figure 9).

[0102] Example 9: SP163M shows efficacy against atopic dermatitis Similar to human esophageal epithelial cells, treatment of human keratinocytes with Poly I:C resulted in increased TSLP production and associated increased cell death. Treatment of Poly I:C-induced human keratinocytes with SP163M both reduced TSLP (Figure ​(Figure10A)) and increased cell viability (Figure10B). In this model of skin inflammation, SP163M reduced a key cytokine (TSLP) that regulates allergic responses and protected cells from death.

[0103] In epidermal keratinocyte (HaCat) cells, SP163M treatment inhibited the phosphorylation of IκBα by the proinflammatory cytokine TNF-α in an in vitro model of atopic dermatitis. TNFα treatment caused phosphorylation of IκBα (inhibitor of nuclear factor κB), which in turn activated NFκB. Thus, in a skin inflammation model, SP163M blocked a key inflammatory pathway (Figure 10C).

[0104] In a calcipotriol / ova-induced atopic dermatitis model in mice, treatment with SP163M resulted in significantly lower eosinophil levels per section compared to vehicle controls (p = 0.045). SP163M treatment resulted in reduced eosinophil infiltration compared to A1AT administered at a 20-fold higher dose (Figure 11). Thus, SP163M improved the outcome of an atopic dermatitis animal model.

[0105] As shown in Figures 16A-16C, in a model of atopic dermatitis induced by calcipotriol for 14 days, mice treated with SP16 topical solution (1%) show significant amelioration of disease compared to vehicle-treated mice.

[0106] As shown in Figure 17, PAR2 and TSLP are upregulated in skin biopsies of AD mice. Inhibition of these upstream mediators results in attenuation of scratching behavior and inflammation in mice. SP16-treated mice show significant reduction in systemic (serum) and local (ear) TSLP compared to MC903-treated mice. SP16 treatment significantly reduces pruritus compared to vehicle (MC903)-treated mice. SP16-treated animals show significant reduction in downstream TH2-mediated IL-4 release in skin tissue.

[0107] Example 10: SP16 inhibits key allergic inflammatory mediators SP16 reduces several key cytokines involved in allergic responses. Knocking out SPINK7 causes esophageal epithelial cells to release exacerbating levels of TSLP. TSLP is a major mediator of immune cell responses upon activation to insults such as allergens. Similarly, CCL26 is a chemokine involved in allergen-induced eosinophil activation and is also upregulated upon SPINK7 knockout. In primary SPINK7 knockout esophageal cells, SP16 (and an SP16 analog targeting LRP1, 7G) are effective in significantly reducing poly I:C-mediated TSLP and CCL26 release (Figure 12).

[0108] The OVA-induced allergic inflammation model is a widely used eosinophil-mediated allergic disease model. As shown in Figures 13-14, using the OVA-induced allergy model in mice, SP16 significantly reduced cytokines in bronchoalveolar lavage fluid (BALF).

[0109] Using an OVA-induced allergic inflammation model, the effect of SP16 treatment on eosinophil infiltration in mice was evaluated. The number of eosinophils in bronchoalveolar lavage fluid was quantified by flow cytometry. Figure 15 shows that SP16 significantly (p = 0.04) reduced eosinophil infiltration into the lungs.

[0110] Additional Examples Use of serpin peptides in reducing inflammation in subjects with LRP1 or TSLP-related diseases or conditions In some embodiments, the technology includes the use of serpin peptides in reducing inflammation in subjects with diseases or conditions associated with LRP1 or TSLP. In some aspects, the technology includes administering any of the serpin peptides described in the present disclosure.

[0111] In some aspects, the use of serpin peptides in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP includes: The present invention includes administering a serpin peptide comprising an amino acid sequence selected from the group consisting of: TIFF2025503607000021.tif48165, wherein the serpin peptide is administered to the subject to reduce inflammation associated with a disease or condition associated with LRP1 or TSLP.

[0112] In some aspects, the use of serpin peptides in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP includes administering a serpin peptide comprising SP16 peptide (SEQ ID NO: 2) or SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP163M peptide (SEQ ID NO: 35).

[0113] In some aspects, the use of a serpin peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a serpin peptide comprising the amino acid sequence X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In some aspects, the serpin peptide is Contains sequence TIFF2025503607000022.tif19164.

[0114] In some aspects, the use of serpin peptides in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a serpin peptide comprising the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO: 20), where X1 is V or L, X2 is R or F, X3 is R or K, X4 is M, Nle or I, Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is a sequence of any 5 amino acids. In some aspects, the serpin peptide comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO:21), where X1 is V or L, X2 is F or R, X3 is K or R, X4 is V, L or M, X5 is a sequence of any 5 amino acids, Z1 is any amino acid, Z2 is a sequence of any 2 amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0115] In some aspects, the use of a serpin peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP includes administering a serpin peptide comprising an amino acid sequence of the serpin peptide comprising the sequence of SEQ ID NO: 35. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 2. In some aspects, the N-terminus of the serpin peptide is acetylated. In some aspects, the C-terminus of the serpin peptide is amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In some aspects, the one or more other peptides are different from the serpin peptide. In some aspects, the fusion peptide or fusion protein includes the serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the technology includes a pharmaceutical composition including a serpin peptide and a pharma- ceutically effective carrier.

[0116] In some embodiments, the use of a serpin peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering the serpin peptide at a therapeutically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a therapeutically effective dose. In some aspects, the serpin peptide is administered at a clinically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a clinically effective dose. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human. In some aspects, administration is by oral, parenteral, intradermal, transdermal, topical, or intranasal administration. In some aspects, the serpin peptide is administered as a single dose. In some aspects, the serpin peptide is administered by topical administration.

[0117] In some aspects, the use of serpin peptide in reducing inflammation in a subject with a disease or condition associated with LRP1 or TSLP comprises administering serpin peptide, wherein the disease or condition is caused by A. alternata. In some aspects, the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia. In some aspects, the disease or condition is rhinitis. In some aspects, the disease or condition is asthma. In some aspects, the disease or condition is dermatitis. In some aspects, the disease or condition is esophageal eosinophilia. In some aspects, the disease or condition is acute or neuropathic pain. In some aspects, the disease or condition is acute nociceptive, inflammatory, or neuropathic pain. In certain embodiments, the disease or condition is EDD. In some aspects, the disease or condition is EoE, eosinophilic asthma, atopic dermatitis, nasal polyps, chronic spontaneous urticaria, and pruritus. In some aspects, the disease or condition is atopic dermatitis. In some aspects, the disease or condition is pruritus. In some aspects, the disease or condition is an allergic reaction. In some aspects, the disease or condition is allergic inflammation. In some aspects, the disease or condition is an eosinophilic allergic disease. In some aspects, the disease or condition is caused by TH2-type inflammatory cytokines.

[0118] Use of Serpin Peptides in Treating Acute or Neuropathic, Nociceptive, or Inflammatory Pain - Patent application In some embodiments, the technology includes the use of serpin peptides in the treatment of diseases related to LRP1 or TSLP, wherein the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.In an aspect, the technology includes the use of any of the serpin peptides described in this application for the treatment of acute or neuropathic pain, nociceptive pain, or inflammatory pain.

[0119] In some aspects, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes: TIFF2025503607000023.tif49164, wherein the serpin peptide is administered to a subject in need thereof to treat a disease or condition associated with LRP1 or TSLP, and wherein the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.

[0120] In some aspects, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering a serpin peptide comprising SP16 peptide (SEQ ID NO: 2) or SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with SP163M peptide (SEQ ID NO: 35).

[0121] In some aspects, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering a serpin peptide comprising the amino acid sequence X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In some aspects, the serpin peptide is Contains sequence TIFF2025503607000024.tif19170.

[0122] In some aspects, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering a serpin peptide comprising the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO: 20), where X1 is V or L, X2 is R or F, X3 is R or K, X4 is M, Nle or I, Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is any sequence of 5 amino acids. In some aspects, the serpin peptide comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO:21), where X1 is V or L, X2 is F or R, X3 is K or R, X4 is V, L or M, X5 is a sequence of any 5 amino acids, Z1 is any amino acid, Z2 is a sequence of any 2 amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0123] In some aspects, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering a serpin peptide comprising the sequence of SEQ ID NO: 35. In some aspects, the serpin peptide comprises the sequence of SEQ ID NO: 2. In some aspects, the N-terminus of the serpin peptide is acetylated. In some aspects, the C-terminus of the serpin peptide is amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In some aspects, the one or more other peptides are different from the serpin peptide. In some aspects, the fusion peptide or fusion protein includes the serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the technology includes a pharmaceutical composition including a serpin peptide and a pharma- ceutically effective carrier.

[0124] In some embodiments, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering the serpin peptide at a therapeutically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a therapeutically effective dose. In some aspects, the serpin peptide is administered at a clinically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a clinically effective dose. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human. In certain embodiments, administration is by oral administration, parenteral administration, intradermal administration, transdermal administration, topical administration, or intranasal administration. In some aspects, the composition is administered as a single dose.

[0125] In some aspects, the use of serpin peptides in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain includes administering a serpin peptide, where the disease or condition is acute or neuropathic pain. In some aspects, the disease or condition is nociceptive pain. In some aspects, the disease or condition is inflammatory pain. In some aspects, administering a serpin peptide reduces pain. In some aspects, administering a serpin peptide prevents or reduces the onset of pain. In some aspects, administering a serpin peptide increases neuronal survival and neurite sprouting.

[0126] Use of Serpin Peptides in Treating Diseases or Conditions Caused by A. Alternata In some embodiments, the technology includes the use of serpin peptides in treating a subject with a disease or condition associated with LRP1 or TSLP, wherein the disease or condition is caused by A. alternata. In some aspects, the technology includes the use of any of the serpin peptides described in this disclosure.

[0127] In some aspects, the use of serpin peptides in treating a disease or condition caused by A. alternata includes: The method includes administering a serpin peptide comprising an amino acid sequence selected from the group consisting of: TIFF2025503607000025.tif48165, wherein the serpin peptide is administered to the subject to treat a disease or condition associated with LRP1 or TSLP, wherein the disease or condition is caused by A. alternata.

[0128] In some aspects, the use of serpin peptides in treating a disease or condition caused by A. alternata includes administering a serpin peptide comprising SP16 peptide (SEQ ID NO: 2) or SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2). In some aspects, the serpin peptide comprises an amino acid sequence that shares at least 90% identity with the SP163M peptide (SEQ ID NO: 35).

[0129] In some aspects, the use of serpin peptides in treating a disease or condition caused by A. alternata includes administering a serpin peptide comprising the amino acid sequence X1-N-X2-PF-X3-X4-X5-X6, where X1 is R or F, X2 is K or R, X3 is V or L, X4 is F, V or M, X5 is L, V or I, and X6 is M, I or Nle. In some aspects, the serpin peptide is Contains sequence TIFF2025503607000026.tif19164.

[0130] In some aspects, the use of serpin peptides in treating a disease or condition caused by A. alternata includes administering a serpin peptide comprising the amino acid sequence of X1-Z1-X2-Z2-X3-Z3-FVFL-X4-Z4 (SEQ ID NO: 20), where X1 is V or L, X2 is R or F, X3 is R or K, X4 is M, Nle or I, Z1 is any amino acid, Z2 is any amino acid, Z3 is any amino acid, and Z4 is a sequence of any 5 amino acids. In some aspects, the serpin peptide comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-FL-Z4-X5 (SEQ ID NO:21), where X1 is V or L, X2 is F or R, X3 is K or R, X4 is V, L or M, X5 is a sequence of any 5 amino acids, Z1 is any amino acid, Z2 is a sequence of any 2 amino acids, Z3 is any amino acid, and Z4 is M, Nle or I.

[0131] In some aspects, the use of a serpin peptide in treating a disease or condition caused by A. alternata includes administering a serpin peptide comprising the sequence of SEQ ID NO: 35. In some aspects, the amino acid sequence of the serpin peptide comprises the sequence of SEQ ID NO: 2. In some aspects, the N-terminus of the serpin peptide is acetylated. In some aspects, the C-terminus of the serpin peptide is amidated. In some aspects, the serpin peptide is fused to one or more other peptides to form a fusion peptide or fusion protein. In some aspects, the one or more other peptides are different from the serpin peptide. In some aspects, the fusion peptide or fusion protein includes the serpin peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. In some aspects, the technology includes a pharmaceutical composition including a serpin peptide and a pharma- ceutically effective carrier.

[0132] In some embodiments, the use of serpin peptides in treating a disease or condition caused by A. alternata includes administering the serpin peptide at a therapeutically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a therapeutically effective dose. In some aspects, the serpin peptide is administered at a clinically effective dose or concentration. In some aspects, the pharmaceutical compound is administered at a clinically effective dose. In some aspects, the serpin peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg. In some aspects, the subject is a human. In some aspects, the administration is by oral, parenteral, intradermal, transdermal, topical, or intranasal administration. In some aspects, the serpin peptide is administered as a single dose.

[0133] In some aspects, the use of serpin peptides in treating a disease or condition caused by A. alternata includes administering a serpin peptide, wherein the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia. In some aspects, the disease or condition is rhinitis. In some aspects, the disease or condition is asthma. In some aspects, the disease or condition is dermatitis. In some aspects, the disease or condition is esophageal eosinophilia. In some aspects, administering the serpin peptide reduces inflammation. In some aspects, administering the serpin peptide reduces eosinophilic inflammation.

[0134] References The references, patents and published patent applications listed below, as well as all references cited in the above specification, are hereby incorporated by reference in their entireties, as if fully set forth herein. TIFF2025503607000027.tif209158TIFF2025503607000028.tif226158TIFF2025503607000029.tif224158TIFF2025503607000030.tif233158TIFF2025503607000031.tif242158TIFF2025503607000032.tif243158TIFF2025503607000033.tif242158TIFF2025503607000034.tif35158

Claims

1. A pharmaceutical composition comprising a serpintide peptide for use in a method of reducing inflammation in a subject, wherein the serpintide peptide comprises an amino acid sequence selected from the group consisting of and the inflammation is associated with a disease or condition associated with LRP1 or TSLP, the pharmaceutical composition.

2. wherein the disease or condition is (a) caused by A. alternata and / or (b) rhinitis, asthma, dermatitis, or esophageal eosinophilia, The pharmaceutical composition according to claim 1.

3. The pharmaceutical composition according to claim 1, wherein the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.

4. wherein the disease or condition is (a) an eosinophil-related disease (EDD), optionally where the EDD is eosinophilic esophagitis (EoE), eosinophilic asthma, atopic dermatitis, nasal polyps, or chronic idiopathic urticaria, (b) atopic dermatitis or pruritus, or (c) an allergic reaction, allergic inflammation, or an allergic disease caused by eosinophils, The pharmaceutical composition according to claim 1.

5. A pharmaceutical composition comprising a serpintide peptide for use in a method of treating a disease or condition associated with LRP1 or TSLP in a subject, wherein the serpintide peptide comprises an amino acid sequence selected from the group consisting of and the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain, the pharmaceutical composition.

6. The pharmaceutical composition according to claim 5, wherein administration of the serpintide peptide in the method results in pain reduction and / or prevents or reduces the onset of pain.

7. A pharmaceutical composition comprising a serpintide peptide for use in a method of treating a subject having a disease or condition associated with LRP1 or TSLP, wherein the serpintide peptide comprises an amino acid sequence selected from the group consisting of and the disease or condition is caused by A. alternata, the pharmaceutical composition.

8. The pharmaceutical composition according to claim 7, wherein the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia. **Claim 9**: The pharmaceutical composition according to claim 7 or 8, wherein the administration of the serpins peptide in the method reduces inflammation and optionally, the administration of the serpins peptide in the method reduces eosinophilic inflammation. **Claim 10** The pharmaceutical composition according to claim 1, 5, or 7, wherein the amino acid sequence of the serpins peptide comprises the sequence of SEQ ID NO:

35. **Claim 11** The pharmaceutical composition according to claim 1, 5, or 7, wherein the N-terminus of the serpins peptide is acetylated and / or the C-terminus of the serpins peptide is amidated. **Claim 12** The serpins peptide is fused to one or more other peptides to form a fusion peptide or fusion protein, optionally the one or more other peptides are different from the serpins peptide, and / or the fusion peptide or fusion protein comprises the serpins peptide and an epitope tag, a half-life extender, or both an epitope tag and a half-life extender. The pharmaceutical composition according to claim 1, 5, or 7. **Claim 13**: (i) In the method, the serpins peptide is administered at a dose of 0.001 mg / kg to 5 mg / kg; (ii) In the method, the serpins peptide is administered by oral administration, parenteral administration, intradermal administration, transdermal administration, topical administration, or intranasal administration; (iii) In the method, the serpins peptide is administered as a single dose. The pharmaceutical composition according to claim 1, 5, or 7. **Claim 14**: The pharmaceutical composition according to claim 13, wherein the administration is topical administration. **Claim 15** The pharmaceutical composition according to claim 1, 5, or 7, wherein the subject is human.

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