WO2023133078A2 - Serpin peptides and methods of using the same - Google Patents

Serpin peptides and methods of using the same Download PDF

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Publication number
WO2023133078A2
WO2023133078A2 PCT/US2022/082669 US2022082669W WO2023133078A2 WO 2023133078 A2 WO2023133078 A2 WO 2023133078A2 US 2022082669 W US2022082669 W US 2022082669W WO 2023133078 A2 WO2023133078 A2 WO 2023133078A2
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seq
peptide
serpin
disease
serpin peptide
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French (fr)
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WO2023133078A3 (en
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Dana AUSTIN
Cohava Gelber
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SERPIN PHARMA LLC
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SERPIN PHARMA LLC
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Priority to JP2024540848A priority Critical patent/JP2025503607A/ja
Priority to CN202280092284.4A priority patent/CN118804758A/zh
Priority to CA3247129A priority patent/CA3247129A1/en
Priority to EP22919262.0A priority patent/EP4460322A4/en
Priority to AU2022431253A priority patent/AU2022431253A1/en
Priority to US18/726,945 priority patent/US20250161419A1/en
Priority to IL314135A priority patent/IL314135A/en
Publication of WO2023133078A2 publication Critical patent/WO2023133078A2/en
Publication of WO2023133078A3 publication Critical patent/WO2023133078A3/en
Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
    • C07K14/8121—Serpins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/55—Protease inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • SERPINs Serine protease inhibitors
  • LRP1 low-density lipoprotein receptor-related protein
  • this technology 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, pruritis, chronic spontaneous urticaria, and the use of a SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP, including administering a SERPIN peptide selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLMIEQNTK (SEQ ID NO: 2), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLWI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31), VKFNKPFVFL(Nle)IEQNTK
  • the amino acid sequence of the SERPIN peptide includes the sequence of SEQ ID NO: 35 or SEQ ID NO: 2.
  • the N-terminus of the SERPIN peptide is acetylated.
  • the C-terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the other peptide is different from the SERPIN peptide.
  • the fusion peptide or fusion protein includes the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human, and the SERPIN peptide is administered by oral administration, parenteral administration, intradermal administration, transdermal administration, topical administration, or intranasal administration.
  • the SERPIN peptide is administered as a single dose.
  • disease or condition is caused by A. alternata.
  • the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia.
  • the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.
  • the disease or condition is an eosinophilic driven disease (EDD), for example, eosinophilic esophagitis (EoE), eosinophilic asthma, atopic dermatitis, nasal polyps, or chronic spontaneous urticaria.
  • the disease or condition is atopic dermatitis or pruritis, and the SERPIN peptide may be administered by topical administration.
  • the disease or condition is an allergic reaction, allergic inflammation, or eosinophilic driven allergic disease.
  • this technology relates to a method of treating or the use of a SERPIN peptide in treating a subject having a disease or condition associated with LRP1 or TSLP, including administering a SERPIN peptide selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLVVI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLVVIR (SEQ ID NO: 41 ), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLWIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO: 46), FVFLM
  • the subject is a human
  • the SERPIN peptide may be administered by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the SERPIN peptide is administered to the subject at a dose of between 0.001 mg/kg and 5 mg/kg, and the SERPIN peptide may be administered as a single dose.
  • administration of the SERPIN peptide results in reduced pain, and/or the administration prevents or reduces the development of pain.
  • this technology relates to a method of treating or the use of a SERPIN peptide in treating a subject having a disease or condition associated with LRP1 or TSLP, including administering a SERPIN peptide comprising an amino acid sequence selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLWI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLVVIR (SEQ ID NO: 41 ), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLWIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO:
  • the amino acid sequence of the SERPIN peptide comprises the sequence of SEQ ID NO: 35, and the N-terminus of the SERPIN peptide may be acetylated and/or the C-terminus of the SERPIN peptide may be amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein, and the other peptide may be different from the SERPIN peptide.
  • the fusion peptide or fusion protein includes the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg, and the subject may be a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration, and the SERPIN peptide may be administered as a single dose.
  • the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia, and administrating the SERPIN peptide may reduce inflammation or eosinophilic inflammation.
  • FIG. 1 demonstrates that SERPINs contain an anti-inflammatory core motif comprising the LRP1 binding sequence.
  • NFKB reporter assay stimulated with LPS
  • the NFKB lowering activity of core SERPIN peptides was lost when truncated too short due to instability of the truncated peptides.
  • the activity was restored upon poly arginine residues stabilizing the peptides.
  • the LRP1 binding site was truncated, the activity could not be rescued by poly arginine flanks.
  • 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 primary adult DRG neurons cultured over time. Cultures were treated with vehicle or SP163M (100 ng ml) daily for 48, 72, and 96 h. Scale bar 500 pm.
  • Figure 2B Representative image of immunofluorescence to detect pil l-Tubulin in primary cultured adult DRG neurons in control and SP163M (240 nM) treated cells after 54 h. Upper panel: scale bar 200 pm; lower panel: scale bar 50 pm. Note extensive neurite length in SP163M treated neurons.
  • Figures 3A-3F show that in PC12 cells, SP163M activated transient cell signaling in an LRP1 dependent manner.
  • 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 SP163M (240 nM) activation of phospho-ERK1/2.
  • Last lane (far right) shows activation of phospho-ERK1/2 by a known LRPI agonist, EI-tPA (12 nM).
  • Equal amounts of protein lysates (20 pg) were loaded per lane. Immunoblot analysis detects phospho- ERK1/2 and total ERK1/2, as a loading control.
  • Figure 3D Immunoblot of LRP1 levels in PC12 cells transfected with non-targeting controls (NTC) or siLRPI for 48 h in PC12 cells.
  • Figure 3E Representative immunoblot of phospho- ERK1/2 activated by SP16 (240 nM) overtime 48 h after transfection with NTC or siLRPI .
  • Figure 3F Immunoblot showing activation of phospho-Akt and phospho-ERK1/2 with vehicle or SP163M (24 or 240 nM) for 10 min and in some wells, pretreated with RAP (150 nM) for 15 min. NGF (0.36 nM) for 10 min served as a cell signaling control. Equal amounts of protein lysates (20 pg) were loaded per lane. Total ERK served as a loading control.
  • Figures 4A-4C show that SP163M modulated both early and late phase of the formalin test.
  • Figure 4A Time-course of formalin (2.5%) induced paw licking in C57BL6 mice. Vehicle or SP163M (0-2 pg/g s.c.) was given 1 h prior to formalin injection into the hind paw.
  • Figures 5A-5C show that systemically administered SP163M and EI-tPA attenuated acute nociception induced by intraplantar capsaicin.
  • Figure 5A Nociceptive related behaviors (time spent licking) increased in both male and female mice after intraplantar injection of capsaicin (20 pg) when compared to vehicle (cyclodextrin: 20%). Data are expressed as mean ⁇ SEM. Kruskal-Wallis test ****p ⁇ .0001.
  • Figures 6A-6E show that systemically administered SP163M treatment blocked the development of mechanical hypersensitivity and inflammatory cell recruitment after PNL.
  • Figure 6A Tactile allodynia developed after PNL and were sustained for 14 days.
  • Figures 7A-7C show that inflammatory cell recruitment and satellite cell activation were reduced by SP163M after PNL.
  • Figure 7A Transverse sections of L4 DRG immunostained for CDU b and GFAP after vehicle or SP163M treatment two days post PNL. Note abundant immunoreactivity (brown) identifying CD11 b (black arrows, upper panels) in between neuronal cell bodies and close to blood vessels or identifying GFAP (black arrows, lower panels) surrounding the neuronal cell bodies in injured vehicle-treated (left) or SP163M -treated DRGs (right). CD11 b and GFAP immunoreactivity in SP163M treated DRGs are minimal. Nuclei are stained with hematoxylin (blue).
  • Figures 8A-8C show that SP163M blocked IL-13 stimulated Stat6 phosphorylation.
  • Figure 8A At 30 minutes post 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 post-treatment (many different experimental replicates shown).
  • Figure 8C shows that the SP163M reduction of phosphorylated STAT6 was dependent on expression of LRP1.
  • LRP1 knockout esophageal EPC2 cell line was created. In the control cells (with LRP1) SP163M was capable of reducing phospho- STAT6 expression; however, in the LRP1 knockout cell line, SP163M was not capable of reducing phosphorylated STAT6.
  • Figures 9A-9B show that SP163M reduced eosinophilic esophagitis in a model of eosinophilic esophagitis using A. alternata as allergen.
  • Figure 9A Representative images of anti-MBP staining of esophageal sections following allergen (A. alternata) or control (saline) challenges.
  • Figure 9B Quantification of eosinophils in the esophagus; data shown are the number of eosinophils per high power field (HPF).
  • Figures 10A-10C show that SP163M inhibited TSLP and reduced cell death in human keratinocytes.
  • Figure 10C HaCat cells were insulted with TNFa and immunoblotted for phospho-lkBa (Ser32). GAPDH served as a loading control and signal normalization.
  • FIG 11 shows that SP163M improved outcomes in an atopic dermatitis animal model. Dermatitis was induced on the skin of BALB/c mice with a series of calcipotriol (MC903) and OVA challenges over an 8-week period. SP163M (100 pg/ mouse), A1AT (2 mg/ mouse) or vehicle (DDW) was given twice weekly starting at 4 weeks. The number of eosinophils infiltrating the epithelial sections were quantified by MBP staining.
  • Figures 12A-12B show that SP16 inhibits key allergic inflammatory mediators.
  • Figure 12A Poly l:C-induced TSLP production by SPINK7 knockout EPC2 cells (Human esophageal epithelial cells). SPINK7 KO cells and control cells were plated in high calcium and high density, for 48 hours before being treated with SP16 or 7G and Poly l:C (5 pg/ml, or untreated) for 8 hours. TSLP production in the supernatant was measured by ELISA.
  • Figure 12B primary esophageal epithelial cells (EPC2) were treated with SP16 and then stimulated with IL-13, a TH2 response mediated cytokine, to induce CCL26. SP16 significantly reduced CCL26 release dose dependently.
  • EPC2 primary esophageal epithelial cells
  • Figure 13A-13C shows that SP16 reduces inflammatory markers in the bronchoalveolar lavage fluid of mice following OVA challenge. Mice were subjected to the OVA-challenge allergic inflammation model and treated with vehicle or SP16 during the challenge phase.
  • the bronchoalveolar lavage fluid (BALF) was analyzed for cytokines via ELISA.
  • Figure 13A SP16 significantly reduces CCL24 following OVA- challenge in sensitized mice.
  • Figure 13B SP16 significantly reduces IL-1 a following OVA-challenge in sensitized mice.
  • Figure 13C SP16 significantly reduces IL-2 following OVA-challenge in sensitized mice.
  • FIG 14 shows that SP16 treatment mediates TH2 driven inflammatory cytokines in OVA model.
  • Mice were subjected to the OVA-challenge allergic inflammation model and treated with vehicle (saline), dexamethasone (1 mg/kg) (pos cont.) or SP16 (50pg), for 4 total treatments during the challenge phase (intranasal OVA, 4 treatments over 8 days).
  • BALF and lung tissue was analyzed for cytokines via ELISA. Cells in the BALF were counted with hemacytometer and total WBC count shown.
  • Figure 15A-15B shows that SP16 reduces eosinophilic infiltration into the lung in OVA model.
  • Figure 15A mice were subjected to the OVA-challenge allergic inflammation model and treated with vehicle, SP16, or A1AT during the challenge phase. The BALF was collected and analyzed for the percent of eosinophils and compared to saline (non-challenged) mice, vehicle OVA induced mice, or SP16 treated OVA induced mice.
  • Figure 15B shows results of a flow cytometry analysis of the BALF from Figure 15A.
  • FIG. 16A-16C shows that SP16 improves outcomes in an AD-like inflammatory skin model.
  • SP16 in 70% ethanol
  • Drug vehicle (70% EtOH) was applied to control animals (MC903) and normal control animals (NC) received EtOH in place of MC903.
  • In-life parameters Dermatitis score ( Figure 16A) and ear volume ( Figure 16B) measured on days 0, 3, 7, 9, 11 and 14.
  • Figure 16C pictures were taken on the last day of the study (Day 15) and a randomly chosen representative mouse from each group is shown.
  • Clinical Dermatitis Score (score 0-4 for each parameter) — erythema, scale/dryness, edema, excoriation/erosion (total score 0-16).
  • the SERPIN peptides are isolated peptides.
  • the SERPIN peptides are synthetic peptides.
  • compositions comprising the SERPIN-derived peptides and methods of using the same to treat a number of conditions where a dysregulated immune response or impaired endocytic function, or diseases in which LRP1 mediation could contribute to pathology, such as in conditions associated with peripheral nerve injury (and resulting pain), and allergic inflammation.
  • a dysregulated immune response or impaired endocytic function or diseases in which LRP1 mediation could contribute to pathology, such as in conditions associated with peripheral nerve injury (and resulting pain), and allergic inflammation.
  • the unexpected regenerative and healing properties of these peptides allow use of the compositions comprising such peptides to new indications, and allow preventive intervention in conditions associated with, for example, tissue injury.
  • LRP1 functions as an endocytic and cell signal transduction receptor and has several ligands that induce specific cell signaling cascades that can contribute to cell survival and anti-inflammatory mechanisms. 5 ' 18 22 25 LRP1 is ubiquitously expressed on many different organs, abundantly in brain, lung, heart, and immune cells. Because of these unique capabilities and wide expression on both tissues and immune cells, it plays a critical role in regulating inflammation, cellular metabolism, and maintaining homeostasis.
  • LRP1 regulates inflammatory signaling pathways, such as NFKB and JNK pathways, that induce the conversion of pro-inflammatory (M1 ) macrophages to the anti-inflammatory (M2) macrophage phenotype, regulates the cytokine output, and contributes to effective migration and phagocytosis. 22 26 51 In neutrophils, LRP1 -dependent mechanisms lead to enhanced cell adhesion, chemotaxis, and antibacterial effects of these cells, thereby resisting immunosuppression 25 . During acute infection or injury, LRP1 also promotes inflammatory resolution through scavenging PAM PS and DAMPS from dying or injured tissue, to prevent the tissue injury cycle 25 .
  • LRP1 was also shown to mediate autophagy during infection, an important metabolic process recently shown to play an important protective role in a variety of diseases. 4 10 Therefore, because of its multifunctional ability to regulate inflammation, targeting LRP1 has substantial potential to mitigate several aspects of the immune response that contributes to the pathology of several diseases including neurological disorders, infectious diseases, and allergic inflammatory disease.
  • Peripheral nerve injury resulting from metabolic, chemotherapy, or trauma often results in chronic pain.
  • Neuropathic pain is characterized by evoked (allodynia, hyperalgesia) and spontaneous pain-like symptoms. The symptoms may be severe, including paresthesia, tingling, numbness, and burning sensations.
  • Other than short term symptomatic relief few therapeutic options are available and include steroids, local anesthetics, antidepressants, anti-seizure drugs, and opioids, which are reserved for severe pain. All these treatments aim at temporarily reducing pain to manageable levels; however, all can cause side effects and addiction and do not promote healing of damaged nerves. Accordingly, there is an unmet clinical need for novel and innovative pain treatments to prevent the transition from acute to chronic pain.
  • LRP1 agonists are capable of promoting axonal growth in the C NS and are capable of inducing regeneration after spinal cord injury.
  • 53 LRP1 is an endocytic receptor to a diverse number of ligands, including tissue-type plasminogen activator (tPA), matrix metalloproteinase-9 (MM P-9), and activated a2-macroglobulin. 14 These ligands are capable of inducing anti-inflammatory activity, 39 activating the Schwann cell repair program, 21 and transactivation of cell signaling pathways in neurons associated with axonal regeneration.
  • LRP1 requires ligand-binding to activate cell-signaling, however, different ligands elicit distinct 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 with numerous effects on cell physiology that do not involve LRP1-binding. For example, tissue-type plasminogen activator (tPA) binds to LRP1 to promote Schwann cell (SC) survival and migration. 23 Yet, by LRP1- independent activities, tPA elicits pain.
  • tissue-type plasminogen activator binds to LRP1 to promote Schwann cell (SC) survival and migration. 23 Yet, by LRP1- independent activities, tPA elicits pain.
  • EI-tPA promotes survival of human iPSC-derived neural progenitor cells (iNPCs) and transplanted EI-tPA activated iNPCs into rodents with severe spinal cord injury demonstrate improved motor functional recovery. 40 Imbalances in the microenvironment following nerve injury may have severe consequences, including the development of chronic neuropathic pain states. 12 In peripheral nervous system (PNS) injury, both inflammatory cytokines, such as TNFa, IL-6 and IL-1 (3, and antiinflammatory cytokines, such as IL-10, have been shown to play a central role in axon regeneration and repair. 6
  • PNS peripheral nervous system
  • ETD Eosinophilic Driven Diseases
  • EDDs Eosinophilic driven diseases
  • EDDs include EoE, eosinophilic asthma, atopic dermatitis, nasal polyps, and chronic spontaneous urticaria.
  • the peptides disclosed herein can serve as an adjunct treatment to mitigate the acute inflammatory response in patients with ST segment elevation myocardial infarction (STEM I). Additionally, SP163M has been shown to be safe and well tolerated in both Phase I and the ongoing Phase Ila clinical trials.
  • the peptide therapy is unique in that it rebalances dysregulated immune responses and protects tissues from injury without any major immunosuppressive effects. Therefore, it is a safe anti-inflammatory drug that has broad-spectrum utility across a wide variety of immune-mediated diseases.
  • EoE is a chronic, largely type-2 immune mediated allergic inflammatory response associated with esophageal dysfunction and disturbed epithelial barrier function.
  • This food allergen-driven disease is characterized by eosinophil dominated inflammation and type-2 mediated immune responses that lead to esophageal damage.
  • an estimated 150,000 patients (largely children) are currently suffering from this disorder that commonly 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 FDA-approved therapeutics for the treatment of EoE, and management of the disease consists of diet restrictions, proton pump inhibitors, and corticosteroids.
  • the current standard-of-care therapies do not adequately address the immune dysregulation that occurs in EoE and other allergic inflammatory diseases, such as asthma or atopic dermatitis.
  • EoE is characterized by a high number of eosinophils, proteases, cathelicidin, serine proteases, including the kallikreins (KLK5), as well thymic stromal lymphopoietin (TSLP) — a cytokine and master regulator of allergic type-2 inflammatory responses in the local environment.
  • KLK5 kallikreins
  • TSLP thymic stromal lymphopoietin
  • 41 In esophageal epithelial cells, a loss of the function of a serine peptidase inhibitor, Kazal type 7-SPINK7, results in uncontrolled protease activity, release of pro-inflammatory cytokines, such as TNFa, CCL2, GM-CSF, IL-8, and CXCL10, and inflammation.
  • LRP1 deletion specifically of CD11b and CD11c dendritic cells in mice results in heightened allergic inflammatory response in an allergic airway disease model. 29 Mice with LRP1 deletion had increased antigen uptake and suffered increased eosinophilic inflammation, allergic sensitization, Th2 mediated cytokine production, and a reduction in T-regulatory cells. 29 Therefore, LRP1 could aid in maintaining homeostasis of proteases/inhibitors in the esophageal environment, mediating the TH2 responses and inhibiting inflammatory signaling pathways (NFKB, JNK) resulting in repair of esophageal dysfunction.
  • NFKB inflammatory signaling pathways
  • LRP1 is an endocytic scavenger receptor for numerous ligands which exert biologically distinct functions.
  • the LRP1 protein consists of a smaller (85kD, [3 chain) intracellular fragment which spans the cell membrane, non-covalently attached to an extracellular fragment (515kD, a chain) which consists of ligand-binding-type repeats, responsible for the majority of ligand binding.
  • LRP1 In addition to its ability to mediate the endocytosis forvarious lipoproteins, protease/inhibitor complexes, viruses, matrix proteins, and growth factors via its extracellular domain, LRP1 interacts with various scaffolding and signaling proteins via its intracellular domain to mediate cell signaling. Due to the multifunctional capability (both endocytic and cell signaling control) of LRP1 , it is implicated in a variety of biological functions, including cell growth/survival, homeostasis, cell metabolism, cytokine regulation, and trafficking foreign antigens. Therefore, LRP1 is implicated to play a role in a variety of diseases.
  • the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 60% identity to SP16. In certain embodiments, the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 65% identity to SP16. In certain embodiments, the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 70% identity to SP16. In certain embodiments, the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 75% identity to SP16. In certain embodiments, the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 80% identity to SP16.
  • the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 85% identity to SP16. In certain embodiments, the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 90% identity to SP16. In certain embodiments, the SERPIN peptide disclosed herein is an analog or derivative of SP16 peptide, sharing at least 95% identity to SP16.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of a core sequence FNKPFVFLM (SEQ ID NO: 1 ) of the SP16 peptide which has a sequence of VKFNKPFVFLMIEQNTK (SEQ ID NO: 2).
  • the core sequence includes the LRP1 binding site having a sequence of FVFLM.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of a core binding motif having a sequence of X1-N-X2-P-F-X3-X4-X5-X6, wherein 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.
  • the binding motif has a sequence of FNKPFVFLM (SEQ ID NO: 1 ), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLVVI (SEQ ID NO: 6), FNRPFLW[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • the binding motif has a sequence of SEQ ID NO: 1.
  • the binding motif has a sequence of SEQ ID NO: 5.
  • the binding motif has a sequence of SEQ ID NO: 6.
  • the binding motif has a sequence of SEQ ID NO: 7.
  • the binding motif has a sequence of SEQ ID NO: 8. In certain embodiments, the binding motif has a 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 a sequence of F-X3-X4-X5-X6, wherein X3 is
  • the LRP1 binding site has a sequence of FVFLM (SEQ ID NO: 3), FVFL[Nle] (SEQ ID NO: 10), FLWI (SEQ ID NO: 11), FLW[Nle] (SEQ ID NO: 12), FLMII (SEQ ID NO: 13), or FLMI[Nle] (SEQ ID NO: 14).
  • the binding motif has a sequence of SEQ ID NO: 3.
  • the binding motif has a sequence of SEQ ID NO: 10.
  • the binding motif has a sequence of SEQ ID NO: 11.
  • the binding motif has a sequence of SEQ ID NO: 12.
  • the binding motif has a sequence of SEQ ID NO: 13.
  • the binding motif has a sequence of SEQ ID NO: 14.
  • the SERPIN peptides disclosed herein comprise, consist essentially of, or consist of a modified core binding motif, by adding a flanking sequence comprising one or more basic amino acids, an arginine, or both of one or more basic amino acids and an arginine to either or both sides of the core binding motif.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of an amino acid sequence of Z1-R-X1-N-X2-P-F-X3-X4-X5-X6-R-Z2, wherein 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 between 1 and 3, between 1 and 5, between 1 and 6, between 1 and 7, between 1 and 8, between 1 and 9, or between 1 and 10 basic amino acids. In certain embodiments, Z1 is 1 basic amino acid.
  • 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 certain embodiments, Z2 is
  • Z2 is 6 basic amino acids. In certain embodiments, Z2 is 7 basic amino acids. In certain embodiments, Z2 is 8 basic amino acids. In certain embodiments, Z2 is 9 basic amino acids. In certain embodiments, Z2 is 10 basic amino acids. In certain embodiments, Z1 is between 1 and 3 basic amino acids. In certain embodiments, Z1 is between 1 and 5 basic amino acids. In certain embodiments, Z1 is between 1 and 6 basic amino acids. In certain embodiments, Z1 is between 1 and 7 basic amino acids. In certain embodiments, Z1 is between 1 and 8 basic amino acids. In certain embodiments, Z1 is between 1 and 9 basic amino acids. In certain embodiments, Z1 is between 1 and 10 basic amino acids.
  • Z2 is between 1 and 3 basic amino acids. In certain embodiments, Z2 is between 1 and 5 basic amino acids. In certain embodiments, Z2 is between 1 and 6 basic amino acids. In certain embodiments, Z2 is between 1 and 7 basic amino acids. In certain embodiments, Z2 is between 1 and 8 basic amino acids. In certain embodiments, Z2 is between 1 and 9 basic amino acids. In certain embodiments, Z2 is between 1 and 10 basic amino acids.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of an amino acid sequence of Z1-RFNRPFLWIR-Z2 (SEQ ID NO: 17), Z1-RFNRPFLMIIR-Z2 (SEQ ID NO: 18), or Z1-KFNKPFVFL(Nle)R-Z2 (SEQ ID NO: 19), wherein Z1 and Z2 are independently 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, or between 1 and 3, between 1 and 5, between 1 and 6, between 1 and 7, between 1 and 8, between 1 and 9, or between 1 and 10 basic amino acids.
  • the SERPIN peptide comprises, consists essentially of, or consists of an amino acid sequence of Z1-RFNRPFLWIR-Z2 (SEQ ID NO: 17). In certain embodiments, the SERPIN peptide comprises, consists essentially of, or consists of an amino acid sequence of Z1-RFNRPFLWIR-Z2 (SEQ ID NO: 18). In certain embodiments, the SERPIN peptide comprises, consists essentially of, or consists of an amino acid sequence of Z1-RFNRPFLVVIR-Z2 (SEQ ID NO: 19). 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
  • 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 certain embodiments, Z2 is 5 basic amino acids. In certain embodiments, Z2 is 6 basic amino acids. In certain embodiments, Z2 is 7 basic amino acids. In certain embodiments, Z2 is 8 basic amino acids. In certain embodiments, Z2 is 9 basic amino acids. In certain embodiments, Z2 is 10 basic amino acids.
  • Z1 is between 1 and 3 basic amino acids. In certain embodiments, Z1 is between 1 and 5 basic amino acids. In certain embodiments, Z1 is between 1 and 6 basic amino acids. In certain embodiments, Z1 is between 1 and 7 basic amino acids. In certain embodiments, Z1 is between 1 and 8 basic amino acids. In certain embodiments, Z1 is between 1 and 9 basic amino acids. In certain embodiments, Z1 is between 1 and 10 basic amino acids. In certain embodiments, Z2 is between 1 and 3 basic amino acids. In certain embodiments, Z2 is between 1 and 5 basic amino acids. In certain embodiments, Z2 is between 1 and 6 basic amino acids. In certain embodiments, Z2 is between 1 and 7 basic amino acids. In certain embodiments, Z2 is between 1 and 8 basic amino acids. In certain embodiments, Z2 is between 1 and 9 basic amino acids. In certain embodiments, Z2 is between 1 and 10 basic amino acids.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F- V-F-L-X4-Z4 (SEQ ID NO: 20), wherein:
  • this peptide comprises, consists essentially of, or consists of 20 or fewer amino acids.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F- X4-F-L-Z4-X5 (SEQ ID NO: 21 ), wherein:
  • X1 is V or L
  • 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
  • Z3 is any amino acid
  • Z4 is M, Nle, or I.
  • this peptide comprises, consists essentially of, or consists of 20 or fewer amino acids.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 35. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 25. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 29. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 31. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 40.
  • the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 41. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 42. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 43. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 44. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 45. In certain embodiments, the SERPIN peptide disclosed herein comprises, consists essentially of, or consists of the sequence of SEQ ID NO: 46.
  • the SERPIN peptide disclosed herein has a size of between 5 and 30 amino acids.
  • the SERPIN peptide 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
  • the SERPIN peptide may have a size of 5 amino acids. In some embodiments, the SERPIN peptide may have a size of 6 amino acids. In some embodiments, the SERPIN peptide may have a size of 7 amino acids. In some embodiments, the SERPIN peptide may have a size of 8 amino acids. In some embodiments, the SERPIN peptide may have a size of 9 amino acids. In some embodiments, the SERPIN peptide may have a size of 10 amino acids. In some embodiments, the SERPIN peptide may have a size of 11 amino acids. In some embodiments, the SERPIN peptide may have a size of 12 amino acids.
  • the SERPIN peptide may have a size of 13 amino acids. In some embodiments, the SERPIN peptide may have a size of 14 amino acids. In some embodiments, the SERPIN peptide may have a size of 15 amino acids. In some embodiments, the SERPIN peptide may have a size of 16 amino acids. In some embodiments, the SERPIN peptide may have a size of 17 amino acids. In some embodiments, the SERPIN peptide may have a size of 18 amino acids. In some embodiments, the SERPIN peptide may have a size of 19 amino acids. In some embodiments, the SERPIN peptide may have a size of 20 amino acids.
  • the SERPIN peptide may have a size of 21 amino acids. In some embodiments, the SERPIN peptide may have a size of 22 amino acids. In some embodiments, the SERPIN peptide may have a size of 23 amino acids. In some embodiments, the SERPIN peptide may have a size of 24 amino acids. In some embodiments, the SERPIN peptide may have a size of 25 amino acids. In some embodiments, the SERPIN peptide may have a size of 26 amino acids. In some embodiments, the SERPIN peptide may have a size of 27 amino acids. In some embodiments, the SERPIN peptide may have a size of 28 amino acids.
  • the SERPIN peptide may have a size of 29 amino acids. In some embodiments, the SERPIN peptide may have a size of 30 amino acids. In some embodiments, the SERPIN peptide disclosed herein has a size of 20 amino acids or less.
  • a longer peptide may have a decreased solubility, whereas a shorter peptide may have decreased stability.
  • various modifications can be made to improve stability, such as adding poly R sequences or other flanking sequences and forming a fusion protein.
  • the SERPIN peptides include analogues or derivatives thereof.
  • the native sequence of the SERPIN peptides can be modified to enhance plasma stability and result in an increased binding affinity to the peptide's cognate receptor.
  • the SERPIN peptides disclosed herein can be further modified to extend the shelf life and/or bioavailability using one or more non-natural peptide bonds or amino acids or by attaching to the peptide functional groups such as polyethylene glycol (PEG).
  • PEG polyethylene glycol
  • the SERPIN peptides disclosed herein are modified by adding one or more amino acid residues such as arginine at either or both ends.
  • 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.
  • the SERPIN peptides disclosed herein can have an N-terminus or C-terminus with additional or modified functional groups.
  • one or both of the N-terminus and C-terminus of the SERPIN peptide may be amidated.
  • the C-terminus of the SERPIN peptide may be amidated.
  • the N-terminus of the SERPIN peptide may be amidated.
  • one or both of the N-terminus and C-terminus of the SERPIN peptide may be acetylated.
  • the N-terminus of the SERPIN peptide may be acetylated.
  • the SERPIN peptide disclosed herein may comprise a sequence selected from a group consisting of Ac- VKFNKPFVFL(Nle)IEQNTK (N-terminal acetylated SEQ ID NO: 35), Ac-VKFNKPFVFLM (N-terminal acetylated SEQ ID NO: 25), Ac-LRFNRPFLVVI (N-terminal acetylated SEQ ID NO: 29), Ac-VRFNRPFLMII (N-terminal acetylated SEQ ID NO: 31 ), Ac- VKFNKPFVFL(Nle) (N-terminal acetylated SEQ ID NO: 40), Ac-RFNRPFLWIR (N- terminal acetylated SEQ ID NO: 41), Ac-RFNRPFLMIIR (N-terminal acetylated SEQ ID
  • the C-terminus of the SERPIN peptide may be amidated.
  • the SERPIN peptide disclosed herein may comprise a sequence selected from a group consisting of VKFNKPFVFL(Nle)IEQNTK-NH2 (C-terminal amidated SEQ ID NO: 35), VKFNKPFVFLM-NH2 (C-terminal amidated SEQ ID NO: 25), LRFNRPFLVVI-NH2 (C- terminal amidated SEQ ID NO: 29), VRFNRPFLMII-NH2 (C-terminal amidated SEQ ID NO: 31 ), VKFNKPFVFL(Nle)-NH2 (C-terminal amidated SEQ ID NO: 40), RFNRPFLWIR-NH2 (C-terminal amidated SEQ ID NO: 41), RFNRPFLMIIR-NH2 (C- terminal amidated SEQ ID NO: 42), RFNKPFVFL(Nle)R-NH2 (C-terminal amidated SEQ ID NO: 35), VKFNKPFVFLM-
  • the C-terminus of the SERPIN peptide may be amidated and the N-terminus may be acetylated.
  • the SERPIN peptide disclosed herein may comprise a sequence selected from a group consisting of 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-LRFNRPFLWI-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 acetylated SEQ ID NO
  • the SERPIN peptides disclosed herein are fused to one or more other peptides to form a fusion peptide or fusion protein.
  • 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 of an epitope tag and a half-life extender.
  • the peptide comprises one or more D-amino acids, that is, one or more amino acids of the peptide have a D-configuration.
  • this disclosure relates to a pharmaceutical composition
  • a pharmaceutical composition comprising, consisting essentially of, or consisting of an effective amount of one or more SERPIN peptides or fusion peptides disclosed herein.
  • the pharmaceutical composition further comprises one or more additional therapeutic agents, which are not the SERPIN peptides disclosed herein.
  • the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, additive, preservative, or a combination thereof.
  • acceptable carriers include physiologically acceptable solutions, such as sterile saline and sterile buffered saline.
  • an effective amount refers to an amount of a composition that produces a desired effect.
  • An effective amount of a composition may be used to produce a prophylactic or therapeutic effect in a subject, such as preventing or treating a target condition, alleviating symptoms associated with the condition, or producing a desired physiological effect.
  • the effective amount of a composition is a "therapeutically effective amount,” “therapeutically effective concentration” or “therapeutically effective dose.”
  • the precise effective amount or therapeutically effective amount is an amount of the composition that will yield the most effective results in terms of efficacy of treatment in a given subject or population of cells.
  • composition including activity, pharmacokinetics, pharmacodynamics, and bioavailability
  • physiological condition of the subject including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage, and type of medication
  • an effective or therapeutically effective amount may vary depending on whether the composition is administered alone or in combination with another composition, drug, therapy, or other therapeutic method or modality.
  • 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 is shown to be effective in clinical trials or is predicted to be effective based on early phase or pre-clinical trials.
  • Remington The Science and Practice of Pharmacy, 21 st Edition, Univ, of Sciences in Philadelphia (USIP), Lippincott Williams & Wilkins, Philadelphia, PA, 2005.
  • the peptides or the pharmaceutical compositions disclosed herein may be formulated for oral administration, parenteral administration, such as intravenous administration, intramuscular administration, subcutaneous administration (bolus injection or through a device such as an infusion pump), intradermal administration, transderm al administration, topical administration, and intranasal administration.
  • parenteral administration such as intravenous administration, intramuscular administration, subcutaneous administration (bolus injection or through a device such as an infusion pump), intradermal administration, transderm al administration, topical administration, and intranasal administration.
  • a subcutaneous infusion pump can be used for delivery of the peptides or the pharmaceutical compositions disclosed herein.
  • the peptides or the pharmaceutical compositions may be administered more than once. More specifically, after the initial administration, one or more additional doses may be given as a booster.
  • the SERPIN peptides or the pharmaceutical compositions disclosed herein have various functions.
  • disclosed herein is a method of treating a subject in need thereof an effective amount of one or more SERPIN peptides, fusion peptides, or the pharmaceutical compositions disclose herein.
  • the subject suffers from a disease or condition associated with LRP1 or TSLP.
  • the subject suffers from acute neuropathic pain, such as acute nociceptive, inflammatory, and neuropathic pain.
  • the subject suffers from an EDD, such as EoE, eosinophilic asthma, atopic dermatitis, nasal polyps, and chronic spontaneous urticaria.
  • the subject suffers from an allergic disease, allergic inflammation, or an eosinophil driven allergic disease.
  • treating or “treatment” of a condition may refer to preventing the condition, slowing the onset or rate of development of the condition, reducing the risk of developing the condition, preventing or delaying the development of symptoms associated with the condition, reducing or ending symptoms associated with the condition, generating a complete or partial regression of the condition, or some combination thereof. Treatment may also mean a prophylactic or preventative treatment of a condition.
  • the term "subject" is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject has not received any prior treatment with serine protease inhibitors, such as alpha-1 -antitrypsin treatment, before the treatment with the peptides disclosed herein.
  • serine protease inhibitors such as alpha-1 -antitrypsin treatment
  • the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions can be adm inistered at a dose from about 0.001 mg/kg to about 4 mg/kg in humans.
  • a suitable dose can be selected accordingly. For example, for acute indications, fewer treatments with a higher dose in each treatment are administered; while for chronic indications requiring frequent and long-term treatment, a lower dose in each treatment is administered.
  • a very low dose of the SERPIN peptides such as SP16 and SP163M is required.
  • neurons express a very high density of LRP1. In vivo and in vitro studies demonstrated a significant effect with a low dose of 0.05 pg.
  • the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions is administered subcutaneously.
  • a SERPIN peptide such as SP16 or SP163M, is administered subcutaneously to a human subject at a dose of between 0.05 mg/kg and 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.
  • the SERPIN peptide is administered at a dose of 0.05 mg/kg.
  • the SERPIN peptide is administered at a dose of 0.1 mg/kg. In some embodiments, the SERPIN peptide is administered at a dose of 0.15 mg/kg. In some embodiments, the SERPIN peptide is administered at a dose of 0.2 mg/kg. In some embodiments, the SERPIN peptide is 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.
  • 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, a SERPIN peptide such as SP 16 or SP163M, is administered subcutaneously to a human subject at a dose of 0.2 mg/kg or 0.4 mg/kg.
  • a SERPIN peptide such as SP16 or SP163M
  • a human subject is administered orally to a human subject at a dose between 1 mg and 150 mg, for example, at a dose of 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.
  • a SERPIN peptide such as SP16 or SP163M
  • a SERPIN peptide is administered orally to a human subject at a dose of 5 mg, 25 mg, or 100 mg.
  • a SERPIN peptide, such as SP16 or SP163M is administered locally, for example, by a transderm al patch, optionally with pulsatile delivery, at a dose between 0.05 mg/kg and 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.
  • a single dose of the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions is administered.
  • the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions is administered as the sole therapeutic agent.
  • the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions is administered in combination with a secondary therapeutic agent.
  • the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions is administered in a pulsatile mode or a continuous mode. In some embodiments, the SERPIN peptides and fusions thereof or pharmaceutical compositions comprising the peptides or fusions is administered via a transderm al patch, an inhaler, or an intranasal device.
  • the present technology includes a method of reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP.
  • the method comprises administering any SERPIN peptide described in this disclosure to a subject in need thereof.
  • the SERPIN peptide that is administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises an amino acid sequence selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLMIEQNTK (SEQ ID NO: 2), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLVVI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLWIR (SEQ ID NO: 41), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLVVIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO: 46), FVFLM
  • the SERPIN peptide that is administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP com prises an amino acid sequence of X1-N-X2-P-F-X3-X4-X5-X6, wherein 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.
  • the SERPIN peptide comprises a sequence of FNKPFVFLM (SEQ ID NO: 1 ), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLVVI (SEQ ID NO: 6), FNRPFLVV[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • the SERPIN peptide that is administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-V-F-L-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 a sequence of any five amino acids.
  • the SERPIN peptide comprises an amino acid sequence of X1- Z1-X2-Z2-X3-Z3-F-X4-F-L-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 a sequence any five amino acids; Z1 is any amino acid; Z2 is a sequence of any two amino acids; Z3 is any amino acid; and Z4 is M, Nle, or I.
  • the SERPIN peptide that is 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.
  • the amino acid sequence of the SERPIN peptide comprises the sequence of SEQ ID NO: 2.
  • the N-terminus of the SERPIN peptide is acetylated.
  • the C-terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the one or more other peptides is different from the SERPIN peptide.
  • the fusion peptide or fusion protein comprises the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the technology includes a pharmaceutical composition comprising the SERPIN peptide and a pharmaceutically effective carrier.
  • the SERPIN peptide that is 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.
  • the pharmaceutical compound is administered at a therapeutically effective dose.
  • the SERPIN peptide is administered at a clinically effective dose or concentration.
  • the pharmaceutical compound is administered at a clinically effective dose.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the composition is administered as a single dose.
  • the SERPIN peptide is administered by topical administration.
  • the SERPIN peptide is administered by oral administration.
  • the SERPIN peptide that is administered to reduce inflammation in a subject having a disease or condition associated with LRP1 or TSLP, wherein the disease or condition is caused by A. alternata.
  • the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia.
  • the disease or condition is rhinitis.
  • the disease or condition is asthma.
  • the disease or condition is dermatitis.
  • the disease or condition is esophageal eosinophilia.
  • the disease or condition is an acute or neuropathic pain.
  • the disease or condition is acute nociceptive, inflammatory, or neuropathic pain.
  • the disease or condition is an EDD.
  • the disease or condition is EoE, eosinophilic asthma, atopic dermatitis, nasal polyps, or chronic spontaneous urticaria.
  • the disease or condition is atopic dermatitis.
  • the disease or condition is pruritis.
  • the disease or condition is an allergic reaction.
  • the disease or condition is allergic inflammation.
  • the disease or condition is an eosinophilic driven allergic disease.
  • the disease or condition is caused by TH2 driven inflammatory cytokines.
  • the present technology includes a method of treating a subject having a disease or condition associated with LRP1 or TSLP, where the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.
  • the present technology includes a method of treating a subject having a disease or condition associated with LRP1 , where the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.
  • the method comprises administering any SERPIN peptide described in this disclosure to a subject in need thereof.
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide that comprises an amino acid sequence selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLMIEQNTK (SEQ ID NO: 2), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLVVI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLWIR (SEQ ID NO: 41), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLWIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO: 46), FVF
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide that comprises the SP16 peptide (SEQ ID NO: 2) or the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 90% identity with the SP163M peptide (SEQ ID NO: 35).
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide that comprises an amino acid sequence of X1-N-X2-P-F-X3-X4-X5-X6, wherein 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.
  • the SERPIN peptide comprises a sequence of FNKPFVFLM (SEQ ID NO: 1 ), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLVVI (SEQ ID NO: 6), FNRPFLW[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide comprising an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-V-F-L-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 a sequence any five amino acids.
  • the SERPIN peptide comprises of an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4-F-L-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 a sequence any five amino acids; Z1 is any amino acid; Z2 is a sequence of any two amino acids; Z3 is any amino acid; and Z4 is M, Nle, or I.
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide that comprises the sequence of SEQ ID NO: 35.
  • the amino acid sequence of the SERPIN peptide comprises the sequence of SEQ ID NO: 2.
  • the N-terminus of the SERPIN peptide is acetylated.
  • the C-terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the one or more other peptides is different from the SERPIN peptide.
  • the fusion peptide or fusion protein comprises the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the technology includes a pharmaceutical composition comprising the SERPIN peptide and a pharmaceutically effective carrier.
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering the SERPIN peptide at a therapeutically effective dose or concentration.
  • the pharmaceutical compound is administered at a therapeutically effective dose.
  • the SERPIN peptide is administered at a clinically effective dose or concentration.
  • the pharmaceutical compound is administered at a clinically effective dose.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the composition is administered as a single dose.
  • the SERPIN peptide is administered by topical administration.
  • the SERPIN peptide is administered by oral administration.
  • the method of treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering the SERPIN peptide to treat a disease or condition associated with LRP1.
  • the disease or condition is acute or neuropathic pain.
  • the disease or condition is nociceptive pain.
  • the disease or condition is inflammatory pain.
  • administering the SERPIN peptide results in reduced pain.
  • administering the SERPIN peptide prevents or reduces the development of pain.
  • administering the SERPIN peptide results in increased neuronal survival and neurite sprouting.
  • the present technology includes a method of treating a subject having a disease or condition associated with LRP1 or TSLP, where the disease or condition is caused by A. alternata.
  • the present technology includes a method of treating a subject having a disease or condition associated with TSLP, where the disease or condition is caused by A. alternata.
  • the method comprises administering any SERPIN peptide described in this disclosure to a subject having a disease or condition caused by A. alternata.
  • the method of treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising an amino acid sequence selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLMIEQNTK (SEQ ID NO: 2), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLWI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLWIR (SEQ ID NO: 41), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLVVIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO: 46), FVFLM (SEQ ID NO: 35), VKF
  • the method of treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising the SP16 peptide (SEQ ID NO: 2) or the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 90% identity with the SP163M peptide (SEQ ID NO: 35).
  • the method of treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising an amino acid sequence of X1-N-X2-P-F-X3-X4-X5-X6, wherein 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.
  • the SERPIN peptide comprises a sequence of FNKPFVFLM (SEQ ID NO: 1), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLWI (SEQ ID NO: 6), FNRPFLVV[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • the method of treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-V-F-L-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 a sequence any five amino acids.
  • the SERPIN peptide comprises of an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-X4- F-L-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 a sequence any five amino acids; Z1 is any amino acid; Z2 is a sequence of any two amino acids; Z3 is any amino acid; and Z4 is M, Nle, or I.
  • the 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.
  • the amino acid sequence of the SERPIN peptide comprises the sequence of SEQ ID NO: 2.
  • the N-terminus of the SERPIN peptide is acetylated.
  • the C-terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the one or more other peptides is different from the SERPIN peptide.
  • the fusion peptide or fusion protein comprises the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the technology includes a pharmaceutical composition comprising the SERPIN peptide and a pharmaceutically effective carrier.
  • the method of treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide at a therapeutically effective dose or concentration.
  • the pharmaceutical compound is administered at a therapeutically effective dose.
  • the SERPIN peptide is administered at a clinically effective dose or concentration.
  • the pharmaceutical compound is administered at a clinically effective dose.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the SERPIN peptide is administered as a single dose.
  • the 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.
  • the disease or condition is rhinitis.
  • the disease or condition is asthma.
  • the disease or condition is dermatitis.
  • the disease or condition is esophageal eosinophilia.
  • administering the SERPIN peptide reduces inflammation.
  • administering the SERPIN peptide reduces eosinophilic inflammation.
  • mice 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 Institutional Animal Care and Use Committee at University of California, San Diego. All rats and mice were housed with a 12 h:12 h light: dark cycle with ad libitum access to food and water.
  • Reagents SP163M (Ac-VKFNKPFVFLNIelEQNTK-NH2; SEQ ID NO: 35) was provided from Serpin Pharma (Manassas, VA, USA). Briefly, peptides were synthesized by CPC Scientific Inc (Sunnyvale, CA) with purity >95% as verified by high performance liquid chromatography and mass spectroscopy. Recombinant human El- tPA was purchased from Molecular Innovations (Novi, Ml, USA). NGF-[3 was purchased from Sigma (St. Louis, MO, USA). Receptor associated protein was expressed as a glutathione-S-transferase (GST)-fusion protein (GST-RAP).
  • GST glutathione-S-transferase
  • DRG neurons Primary cultured DRG neurons were imaged by phase contrast and the viability of cells was assessed by Trypan blue. Primary DRG neurons were cultured, fixed in 4% paraformaldehyde, and immunofluorescence was performed using a mouse anti-[3lll-tubulin primary antibody (Promega, Madison, Wl, USA; cat#G7121 , 1 :250) and then with Alexa Fluor-488 antimouse antibody (Life Technologies, Carlsbad, CA, USA) as secondary antibody. DRG neurons were imaged at 20x and 40x manually, 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 SP16 and control groups, respectively. Quantification was performed in a blinded manner. For all neurite outgrowth measurements, at least 6 individual experiments were performed in duplicate. [0092] Cell signaling analysis. Rat PC12 cells were purchased from ATCC (CRL-
  • 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 that were precoated 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 adding effectors, 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.
  • SFM serum -free medium
  • cells were pre-incubated with the competitive antagonist of LRP1 , GST-RAP (150 nM) for 15 min or electroporated using the Rat Neuron Nucleofector Amaxa Kit (Lonza Biosciences) and incubated with siRNA to silence LRP1 expression (siLRPI ; M-094191-01-0010, Dharmacon) for 48 h.
  • Control cells were transfected with non-targeting control (NTC) siRNA (NTC; D-001810-10-05, Dharmacon).
  • Cells were rinsed with ice-cold PBS and proteins were extracted in RIPA buffer (20 mM sodium phosphate, 150 mM NaCI, 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, lysates were centrifuged at 15000* g for 5 min, supernatant collected and stored at -20°C. Equal amounts of protein from cell lysates (20 pg), as determined by BCA Protein Assay (ThermoFisher Scientific, Waltham, MA, USA), were subjected to 10% SDS-PAGE and electro-transferred to nitrocellulose membranes.
  • RIPA buffer 20 mM sodium phosphate, 150 mM NaCI, pH 7.4, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1 % SDS
  • protease and phosphatase inhibitors
  • the membranes were blocked with 5% nonfat dried milk and then incubated with primary anti- phospho-AKT (Cell Signaling Technology, Danvers, MA, USA; cat#9271 S; 1 :1000), anti- phospho-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA; cat#9101 S; 1 :1000), anti-LRP1 (Cell Signaling Technology, Danvers, MA, USA; cat#64099S; 1 :1000) or anti total-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA; cat#91012S; 1 :1000).
  • Immunoblots were developed using Radiance, Radiance Q, and Radiance Plus chemiluminescent substrates and imaged using a BioRad ChemiDoc Imaging System (Bio-Rad, Hercules, CA, USA).
  • Capsaicin was dissolved in 20% (2-hydroxypropyl)-[3-cyclodextrin (Sigma-Aldrich, St. Louis, MO, USA) solution. This vehicle concentration solubilized the capsaicin and did not induce a behavioral response when administered alone.
  • the sciatic nerve was exposed at mid-thigh level by separating the biceps femoris and the gluteus superficialis and then carefully cleared of surrounding connective tissue.
  • a 9-0 nylon suture (Ethicon, Inc., Somerville, NJ, USA) was inserted into the nerve and ligated so that the one-third to one-half of the nerve was included.
  • the muscle and skin layers were closed using Reflex? 7 mm stainless steel wound clips (CellPoint Scientific, Inc., Gaithersburg, MD, USA). For behavior testing, mice were acclimated, and baseline tested for one week prior to PNL.
  • Tactile allodynia was tested by applying 0.04 to 4 g Von Frey filaments (Stoelting, Wood Dale IL, USA) to the plantar surface of the ipsilateral hind paw. Filaments were presented in a consecutive fashion either ascending or descending using the up-down method as previously disclosed 60 and modified for mice. 57 61 The filament that caused paw withdrawal 50% of the time (the 50% PWT) was determined. Tactile allodynia was tested on days 2, 4, 9, 11 , and 14 days following PNL. Results were averaged and subjected to statistical analysis. All experiments were performed by an investigator blinded to mouse identity.
  • the membranes were blocked with 5% nonfat dried milk and then incubated with anti- TLR4(CD284)/MD2 (BioLegend, San Diego, CA, USA; cat#117601, 1 :1000), anti-CD11 b (Abeam, Cambridge, MA, USA, cat#Ab1333357) and anti-
  • Primary antibodies were detected with HRP-conjugated species-specific secondary antibodies (Cell Signaling Technology, Danvers, MA, USA; cat#7076S or 4S; 1 :5000).
  • Immunoblots were developed using the SuperSignal West Pico PLUS chemiluminescent substrate (Thermo Fisher Scientific, Waltham, MA, USA), and the Protec Ecomax X-ray film processor. Densitometry analysis was performed using the Image J software (U. S. National Institutes of Health, Bethesda, MD, USA).
  • DRGs were embedded in paraffin. For IHC studies, 4 pm thick DRG tissue sections were immunostained for CDUb (Abeam, Cambridge, MA, USA, cat#Ab1333357; 1 :4500) or GFAP (Dako, Santa Clara, CA, USA; cat#$Z0334; 1 :4000). Slides were immunostained using a Ventana Discovery Ultra (Ventana Medical Systems, Oro, AZ, USA). Antigen retrieval was performed using CC1 (tris-based; pH 8.5) for 40 min at 95°C. The primary antibodies CD11b and GFAP were incubated with the slides for 32 min at 37°C.
  • the secondary antibody, OmniMap anti- HRP (Ventana Medical Systems, Oro, AZ, USA; cat#760-4311 ), was incubated on the sections for 12 min at 37°C. Antibodies were visualized using diaminobenzidine as a chromogen followed by hematoxylin as a counterstain. Slides were rinsed, dehydrated through alcohol and xylene and cover slipped. Light microscopy was performed using a Leica DFC420 microscope with Leica Imaging Software 2.8.1 (Leica Biosystems, Vista, CA, USA).
  • Statistical analysis was performed using GraphPad Prism (GraphPad Prism version 9.1.2 for Mac, GraphPad Software, San Diego, CA, USA). All results are expressed as the mean ⁇ SEM. Comparisons between two groups were performed using two-tailed unpaired T-tests. A non-parametric Mann-Whitney U test was used when the variance in the two populations were significantly different. When greater than two groups were compared, a one-way AN OVA and Tukey's post hoc test was performed or in the case of non-parametric data, the Kruskal-Wallis test was utilized. Measurements of neuropathic pain, in which multiple observations in individual mice over time, were collected and analyzed by repeated-measures ANOVA with a Sidak's post hoc test, p ⁇ .05 was considered statistically significant.
  • a small peptide fragment of the C-terminal end of alpha-1 antitrypsin (the prototypical SERPIN) is capable of binding to LRP1 , exerting potent cell regenerative, tissue protective, and immune-modulatory functions.
  • the naturally occurring degradative C-terminal product of Alpha-1 Antitrypsin (termed C-36) exhibits pro-inflammatory activity similar to lipopolysaccharide in both macrophages and neutrophils. 44
  • the amino acid sequences of the peptides tested are shown in Table 1 below.
  • the core sequences containing the LRP1 binding site for each peptide are shown in bold and underlined.
  • the reporter cells (THP1-XBIue-MD2-CD14 cells) were treated with each peptide (50 pg/ml) before being insulted with LPS (5 ng/ml) and incubated overnight.
  • the NFKB inducible Secreted Embryonic Alkaline Phosphatase (SEAP) was measured in the supernatant and read for absorbance.
  • SEAP NFKB inducible Secreted Embryonic Alkaline Phosphatase
  • various SERPIN peptides sharing a common core motif demonstrated anti-inflammatory activity in NFKB reporter cells when insulted with LPS.
  • LPS insult led to an increase in NFKB activity, and neither the scrambled core control peptide nor the AAT C-36 fragment decreased the NFKB activity.
  • the AAT C-36 fragment showed NFkB inducible properties without the need for LPS.
  • SP16 is capable of reducing NFKB activity
  • the short core peptide SP20 did not have an inhibitory effect.
  • this truncated AAT derived peptide contains the core sequence and LRP1 binding site, it is unstable, and therefore exhibited no activity.
  • SP21 was flanked with triple arginine amino acids on both sides (termed "Poly-R") to obtain SP21
  • the SP21 peptide's stability increased and NFkB inhibition increased as well relative to SP16.
  • the SP16 short core, SP20 peptide was stabilized using triple arginine flanks to obtain SP22, the NFKB activity was significantly reduced.
  • SERPIN 2 short core SP24
  • SERPIN 2 short core poly-R SP26
  • SERPIN 3 short core SP28
  • SERPIN 3 short core Poly-R SP29
  • these peptides all contain an LRP1 binding site, however, when the LRP1 binding site was truncated to obtain SP31 , the anti-inflammatory activity of the peptide was lost and could not be restored with the poly arginine flanks (SP32 peptide). This indicates that SERPINs contain an anti-inflammatory core motif that is dependent on LRP1.
  • 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 in PC12 cells after 10 min that 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 anticipated, and served as a cell signaling control.
  • 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 to screen analgesic and anti-inflammatory pain therapeutics. It was tested whether LRP1 agonism by SP163M regulated pain responses induced by intraplantar formalin ( Figure 4A). SP163M (0.02, 0.2, or 2.0 pg/g) or vehicle was administered one hour prior to paw injection of 2.5% formalin solution. Preemptive administration was initially studied to understand the contribution of primary afferent input in the early phase and is an approach consistent with other formalin test studies testing initial effects of novel opioids or other analgesics. 62 63 Time spent licking was quantified by two blinded observers over 50 min.
  • EI-tPA and SP163M were tested.
  • EI-tPA or SP16 was delivered systemically and one hour preemptively prior to capsaicin injection in both male and female mice.
  • SP163M and EI-tPA blocked the capsaicin induced acute pain-related behavior (Figure 5B).
  • Similar effects of LRP1 agonists were observed in female mice ( Figure 5C).
  • Examples 2-6 demonstrate the robust efficacy of SP163M in three distinct pre-clinical mouse models that includes acute nociceptive, inflammatory, and neuropathic pain. Central to the effect of preventing mechanical hypersensitivity by SP163M, was its potent anti-inflammatory activity in injured peripheral nerves. In these studies, SP163M robustly reduced the early recruitment of inflammatory cells distal to the nerve injury site and in the corresponding L3, L4 DRG early after sciatic nerve ligation. SP163M can delay and/or limit the infiltration of inflammatory cells and thereby regulate pain states.
  • Esophageal EPC2 cells were treated with vehicle (ddw), A1AT or SP163M. Cells were collected at time-points indicated and western blot analysis of phosphospecific STAT6 was analyzed. At 30 minutes post 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 persisted for several hours post-treatment (many different experimental replicates shown) ( Figure 8B). Figure 8C shows that the SP163M reduction of phosphorylated STAT6 was dependent on expression of LRP1. Using CRISPER/CAS9 technology, an LRP1 knockout esophageal EPC2 cell line was created. In the control cells (with LRP1) SP163M was capable of reducing phospho-STAT6 expression; however, in the LRP1 knockout cell line, SP163M was not capable of reducing phosphorylated STAT6.
  • Example 8 SP163M Shows Inhibitory Effect in an Additional Model of Eosinophilic Esophagitis Using A. alternata as Allergen
  • A. alternata is a common airborne mold associated with eosinophilic inflammation and a variety of allergic diseases, such as rhinitis, asthma and dermatitis, and is an established model that induces allergic reaction at the esophagus.
  • the allergen challenges lead to an increase in esophageal eosinophilia.
  • SP163M, A1 AT or vehicle (control) was given twice weekly over the 4-week period for 8 total treatments.
  • esophageal sections were stained for the detection of MBP (eosinophilic marker) and quantitated per high power field.
  • mice treated with SP16 topical solution (1 %) shows a significant improvement in disease vs. vehicle-treated mice.
  • SP16 reduces several key cytokines involved in allergic responses. Upon knockout of SPINK7, esophageal epithelial cells release exacerbated levels of TSLP. TSLP is a key mediator of immune cell responses upon activation to an insult such as allergens. Likewise, CCL26 is a chemokine involved in allergen induced eosinophilic activation, also upregulated upon SPINK7 knockout. In primary SPINK7 knockout esophageal cells, SP16 (and an analog of SP16 that targets LRP1 , 7G) are effective in significantly reducing Poly l:C mediated TSLP and CCL26 release (Figure 12).
  • the ova-induced model of allergic inflammation is a widely used model of eosinophilic driven allergic diseases. As shown in Figs. 13-14, using the ova-induced allergic model in mice, SP16 significantly reduces cytokines in the bronchoalveolar lavage fluid (BALF).
  • BALF bronchoalveolar lavage fluid
  • the technology includes use of a SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP. In some aspects, the technology includes administering any SERPIN peptide described in this disclosure.
  • use of the SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a SERPIN peptide comprising an amino acid sequence selected from the group consisting ofVKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLM (SEQ ID NO: 25), VKFNKPFVFLM IEQNTK (SEQ ID NO: 2), LRFNRPFLWI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLVVIR (SEQ ID NO: 41 ), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLWIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)
  • use of the 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 SP16 peptide (SEQ ID NO: 2) or the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35).
  • 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).
  • use of the SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a SERPIN peptide comprising an amino acid sequence of X1-N-X2-P-F- X3-X4-X5-X6, wherein 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.
  • the SERPIN peptide comprises a sequence of FNKPFVFLM (SEQ ID NO: 1), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLVVI (SEQ ID NO: 6), FNRPFLVV[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • use of the SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a SERPIN peptide comprising an amino acid sequence of X1-Z1-X2-Z2- X3-Z3-F-V-F-L-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 a sequence any five amino acids.
  • the SERPIN peptide comprises of an amino acid sequence of X1 -Z1 -X2-Z2-X3-Z3-F-X4-F-L-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 a sequence of any five amino acids; Z1 is any amino acid; Z2 is a sequence of any two amino acids; Z3 is any amino acid; and Z4 is M, Nle, or I.
  • use of the 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 of the SERPIN peptide comprising the sequence of SEQ ID NO: 35.
  • the amino acid sequence of the SERPIN peptide comprises the sequence of SEQ ID NO: 2.
  • the N-terminus of the SERPIN peptide is acetylated.
  • the C- terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the one or more other peptides is different from the SERPIN peptide.
  • the fusion peptide or fusion protein comprises the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the technology includes a pharmaceutical composition comprising the SERPIN peptide and a pharmaceutically effective carrier.
  • use of the SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a SERPIN peptide at a therapeutically effective dose or concentration.
  • the pharmaceutical compound is administered at a therapeutically effective dose.
  • the SERPIN peptide is administered at a clinically effective dose or concentration.
  • the pharmaceutical compound is administered at a clinically effective dose.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the SERPIN peptide is administered as a single dose. In some aspects, the SERPIN peptide is administered by topical administration.
  • use of the SERPIN peptide in reducing inflammation in a subject having a disease or condition associated with LRP1 or TSLP comprises administering a SERPIN peptide, wherein the disease or condition is caused by A. alternata.
  • the disease or condition is rhinitis, asthma, dermatitis, or esophageal eosinophilia.
  • the disease or condition is rhinitis.
  • the disease or condition is asthma.
  • the disease or condition is dermatitis.
  • the disease or condition is esophageal eosinophilia.
  • the disease or condition is an acute or neuropathic pain.
  • the disease or condition is acute nociceptive, inflammatory, or neuropathic pain.
  • the disease or condition is an EDD.
  • the disease or condition is EoE, eosinophilic asthma, atopic dermatitis, nasal polyps, chronic spontaneous urticaria, and pruritis.
  • the disease or condition is atopic dermatitis.
  • the disease or condition is pruritis.
  • the disease or condition is an allergic reaction.
  • the disease or condition is allergic inflammation.
  • the disease or condition is eosinophilic allergic inflammation.
  • the disease or condition is caused by TH2 driven inflammatory cytokines.
  • the present technology includes use of a SERPIN peptide in the treatment of a disease associated with LRP1 or TSLP, where the disease or condition is acute or neuropathic pain, nociceptive pain, or inflammatory pain.
  • the technology includes use of any of the SERPIN peptides described in this application for the treatment of acute or neuropathic pain, nociceptive pain, or inflammatory pain.
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide comprising an amino acid sequence selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLMIEQNTK (SEQ ID NO: 2), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLWI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLWIR (SEQ ID NO: 41), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLVVIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO: 35),
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide comprising SP16 peptide (SEQ ID NO: 2) or the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35).
  • 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).
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide comprising an amino acid sequence of X1-N-X2-P-F-X3-X4-X5-X6, wherein 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.
  • the SERPIN peptide comprises a sequence of FNKPFVFLM (SEQ ID NO: 1 ), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLWI (SEQ ID NO: 6), FNRPFLVV[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide comprising an amino acid sequence of X1-Z1-X2-Z2-X3-Z3-F-V-F-L- 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 a sequence of any five amino acids.
  • the SERPIN peptide comprises an amino acid sequence of X1 -Z1 -X2-Z2-X3-Z3-F-X4-F-L-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 a sequence any five amino acids; Z1 is any amino acid; Z2 is a sequence of any two amino acids; Z3 is any amino acid; and Z4 is M, Nle, or I.
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide comprising the sequence of SEQ ID NO: 35.
  • the SERPIN peptide comprises the sequence of SEQ ID NO: 2.
  • the N- terminus of the SERPIN peptide is acetylated.
  • the C-terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the one or more other peptides is different from the SERPIN peptide.
  • the fusion peptide or fusion protein comprises the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the technology includes a pharmaceutical composition comprising the SERPIN peptide and a pharmaceutically effective carrier.
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide at a therapeutically effective dose or concentration.
  • the pharmaceutical compound is administered at a therapeutically effective dose.
  • the SERPIN peptide is administered at a clinically effective dose or concentration.
  • the pharmaceutical compound is administered at a clinically effective dose.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the composition is administered as a single dose.
  • the use of the SERPIN peptide in treating acute or neuropathic pain, nociceptive pain, or inflammatory pain comprises administering a SERPIN peptide, wherein the disease or condition is an 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 reduces pain. In some aspects, administering the SERPIN peptide prevents or reduces the development of pain. In some aspects, administering the SERPIN peptide increases neuronal survival and neurite sprouting.
  • the present technology includes use of a SERPIN peptide in treating a subject having a disease or condition associated with LRP1 orTSLP, where the disease or condition is caused by A. alternata.
  • the technology includes use of any SERPIN peptide described in this disclosure.
  • the use of the SERPIN peptide in treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising an amino acid sequence selected from the group consisting of VKFNKPFVFL(Nle)IEQNTK (SEQ ID NO: 35), VKFNKPFVFLMIEQNTK (SEQ ID NO: 2), VKFNKPFVFLM (SEQ ID NO: 25), LRFNRPFLVVI (SEQ ID NO: 29), VRFNRPFLMII (SEQ ID NO: 31 ), VKFNKPFVFL(Nle) (SEQ ID NO: 40), RFNRPFLWIR (SEQ ID NO: 41), RFNRPFLMIIR (SEQ ID NO: 42), RFNKPFVFL(Nle)R (SEQ ID NO: 43), RRRFLVVIRRR (SEQ ID NO: 44), RRRFLMIIRRR (SEQ ID NO: 45), RRRFVFL(Nle)RRR (SEQ ID NO: 46
  • the use of the SERPIN peptide in treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising the SP16 peptide (SEQ ID NO: 2) or the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 95% identity with the SP163M peptide (SEQ ID NO: 35).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 90% identity with the SP16 peptide (SEQ ID NO: 2).
  • the SERPIN peptide comprises an amino acid sequence that shares at least 90% identity with the SP163M peptide (SEQ ID NO: 35).
  • the use of the SERPIN peptide in treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising an amino acid sequence of X1-N-X2-P-F-X3-X4-X5-X6, wherein 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.
  • the SERPIN peptide comprises a sequence of FNKPFVFLM (SEQ ID NO: 1), FNKPFVFL[Nle] (SEQ ID NO: 5), FNRPFLWI (SEQ ID NO: 6), FNRPFLW[Nle] (SEQ ID NO: 7), FNRPFLMII (SEQ ID NO: 8), or FNRPFLVI[Nle] (SEQ ID NO: 9).
  • the use of the SERPIN peptide in treating a disease or condition caused by A alternata comprises administering a SERPIN peptide comprising an amino acid sequence of X1 -Z1 -X2-Z2-X3-Z3-F-V-F-L-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 a sequence any five amino acids.
  • the SERPIN peptide comprises an amino acid sequence of X1-Z1-X2- Z2-X3-Z3-F-X4-F-L-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 a sequence any five amino acids; Z1 is any amino acid; Z2 is a sequence of any two amino acids; Z3 is any amino acid; and Z4 is M, Nle, or I.
  • the use of the SERPIN peptide in treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide comprising the sequence of SEQ ID NO: 35.
  • the amino acid sequence of the SERPIN peptide comprises the sequence of SEQ ID NO: 2.
  • the N- terminus of the SERPIN peptide is acetylated.
  • the C-terminus of the SERPIN peptide is amidated.
  • the SERPIN peptide is fused to one or more other peptides to form a fusion peptide or fusion protein.
  • the one or more other peptides is different from the SERPIN peptide.
  • the fusion peptide or fusion protein comprises the SERPIN peptide and an epitope tag, a half-life extender, or both the epitope tag and the half-life extender.
  • the technology includes a pharmaceutical composition comprising the SERPIN peptide and a pharmaceutically effective carrier.
  • the use of the SERPIN peptide in treating a disease or condition caused by A. alternata comprises administering a SERPIN peptide at a therapeutically effective dose or concentration.
  • the pharmaceutical compound is administered at a therapeutically effective dose.
  • the SERPIN peptide is administered at a clinically effective dose or concentration.
  • the pharmaceutical compound is administered at a clinically effective dose.
  • the SERPIN peptide is administered at a dose of between 0.001 mg/kg and 5 mg/kg.
  • the subject is a human.
  • the administration is by oral administration, parenteral administration, intradermal administration, transderm al administration, topical administration, or intranasal administration.
  • the composition is administered as a single dose.
  • the use of the SERPIN peptide in 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.
  • the disease or condition is rhinitis.
  • the disease or condition is asthma.
  • the disease or condition is dermatitis.
  • the disease or condition is esophageal eosinophilia.
  • administrating the SERPIN peptide reduces inflammation.
  • administering the SERPIN peptide reduces eosinophilic inflammation.
  • Amyloid-beta impairs insulin signaling by accelerating autophagy-lysosom al degradation of LRP-1 and IR-
  • Maheshwari (2008). "Venezuelan equine encephalitis virus infection causes modulation of inflammatory and immune response genes in mouse brain.”
  • Virology Journal 8 197-197. Shi, Y., E. Mantuano, G. Inoue, W. M. Campana, and S. L. Gonias (2009).
  • C-36 peptide a degradation product of alphal - antitrypsin, modulates human monocyte activation through LPS signaling pathways.
  • Tumor necrosis factor receptor- 1 is selectively sequestered into Schwann cell extracellular vesicles where it functions as TN Fa decoy. Glia. 2021. Online ahead of print. doi:10.1002/glia.24098. Orita S., K. Henry, E. Mantuano, et al. "Schwann cell LRP1 regulates Remak bundle ultrastructure and axonal interactions to prevent neuropathic pain.” J Neurosci. 2013:33(13): 5590-5602. doi:10.1523/jneur osci.3342-12.2013. Garcia-Fernandez P., N. Uceyler, and C. Sommer.

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