Anti-par2 antibodies and uses thereof

Abstract

The present disclosure provides antibodies and antigen-binding fragments capable of binding PAR2. In some embodiments, the anti-PAR2 antibodies or antigen-binding fragments thereof bind PAR2 in a pH-dependent manner. The disclosure further provides methods for making and using the antibodies and antigen-binding fragments.

Description

(19) *EP004681726A2* (11) EP 4 681 726 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: 21.01.2026 Bulletin 2026 / 04 (21) Application number: 25204633.9 (22) Date of filing: 16.03.2018 (51) International Patent Classification (IPC): A61K 39 / 00 (2006.01) (52) Cooperative Patent Classification (CPC): C07K 16 / 28; A61P 29 / 00; A61K 2039 / 505; C07K 2317 / 565; C07K 2317 / 76; C07K 2317 / 92; C07K 2317 / 94 (84) Designated Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR Designated Extension States: BA ME Designated Validation States: MA TN (30) Priority: 16.03.2017 US 201762472462 P 02.03.2018 US 201862637766 P (62) Document number(s) of the earlier application(s) in accordance with Art. 76 EPC: 18714185.8 / 3 596 125 (71) Applicant: MedImmune Limited Cambridge CB2 0AA (GB) (72) Inventors: • DOBSON, Claire Cambridge CB21 6GH (GB) • WILLIAMS, Richard Cambridge CB21 6GH (GB) • GURRELL, Ian Cambridge CB21 6GH (GB) • PODICHETTY, Sadhana Cambridge CB21 6GH (GB) • FAIRMAN, David Cambridge CB21 6GH (GB) • THORNTON, Peter Cambridge CB21 6GH (GB) • NEWTON, Philip Cambridge CB21 6GH (GB) (74) Representative: J A Kemp LLP 80 Turnmill Street London EC1M 5QU (GB) Remarks: •The complete document including Reference Table(s) and the Sequence Listing(s) can be downloaded from the EPO website •This application was filed on 25‑09‑2025 as a divisional application to the application mentioned under INID code 62. •Claims filed after the date of filing of the application / after the date of receipt of the divisional application (Rule 68(4) EPC). (54) ANTI‑PAR2 ANTIBODIES AND USES THEREOF (57) The present disclosure provides antibodies and antigen-binding fragments capable of binding PAR2. In some embodiments, the anti-PAR2 antibodies or anti- gen-binding fragments thereof bind PAR2 in a pH-de- pendent manner. The disclosure further provides meth- ods for making and using the antibodies and antigen- binding fragments. EP 4 68 1 72 6 A 2 Processed by Luminess, 75001 PARIS (FR) Description CROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims thebenefit of priority fromU.S.Provisional ApplicationNo. 62 / 472,462, filed onMarch16, 2017 and from U.S. Provisional Application No. 62 / 637,766, filed on March 2, 2018. The foregoing applications are incorporated herein by reference in their entirety. SEQUENCE LISTING

[0002] The instant application contains aSequenceListingwhich has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on March 8, 2018, is named 1848081‑0002‑093-WO1_SL.txt and is 351,668 bytes in size. BACKGROUND OF THE DISCLOSURE

[0003] Chronic pain is a condition that can affect anyone and imposes a burden on patients, health care systems, and economies.Approximately100millionpeople in theUnitedStatessuffer fromchronicpainand the total annual incremental cost of health care due to pain, includingmedical costs and the economic costs of lost time and wages, is estimated to be between $560 and $635 billion dollars (Institute of Medicine of The National Academies, 2011). Yet, in a survey of chronic pain sufferers,more thanhalf felt they had little to no control over their pain (2006Voices ofChronicPainSurvey, American PainFoundation).Paincanbecausedbyavarietyof conditionsanddiseases, fromcancer, todiabetes, toarthritis, andcan be classified into categories: nociceptive, neuropathic, and mixed type pain. Nociceptive pain is defined by stimulation of nerve fibers (e.g. by thermal, mechanical, or chemical stimuli) while neuropathic pain is pain caused by diverse causes such as nerve damage, diseases, and, importantly, inflammation. Inflammation, the process by which organisms recruit immune cells and release immune factors to the site of an injury or infection, can thus be both a helpful process of damage repair and a cause of pain.

[0004] Many treatments for pain inhibit inflammation. Two common classes of anti-inflammatory pain therapeutics are steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs (NSAIDs). Steroidal anti-inflammatory drugs typically suppress prostaglandins and leukotrienes, the products of inflammation. Such drugs are reliable and potent, but carry the risk of severe side effects, including, for example, reduced bone density, weight fluctuations, immune system suppression, and growth / puberty irregularities (Irving, P. M. et al. (2007) Aliment Pharmacol Ther. 26(3):313‑329; Goodman et al. J Am Acad Orthop Surg. 2007 Aug;15(8):450‑60). NSAIDs inhibit cyclooxygenase‑1 and / or 2 (COX‑1 and / or COX‑2), which themselves catalyze the reaction of arachidonic acid into prostaglandins. Chronic pain and inflammation can require prolonged treatment, and prolonged inhibition of COX enzymes can lead to gastrointestinal tract problems, such as gastric bleeding and ulcers. Given the risks associated with such anti-inflammation pain treatments, there is a need for alternative approaches for treating pain.

[0005] G-Protein CoupledReceptors (GPCRs) are a family ofmembrane proteins that share a common structural motif of seven transmembrane domains connecting an N-terminal extracellular domain and a C-terminal intracellular domain (Granier et al, Nat ChemBiol. 2012 Aug; 8(8): 670‑673). GPCRs sense extracellular signals such as photons, hormones, chemokines, etc. and activate intracellular G proteins. Many families of GPCRs exist, such as the Frizzled, Rhodopsin, Secretin, Adhesion, and protease activated receptor (PAR) families (Zhang et al. Nature. 2012 Dec 20; 492(7429): 387‑392; Zhang et al. Mol Cells. 2015 Oct;38(10):836‑42). While the various GPCR families share overall structural features, they exhibit different functions, bind to different ligands, and are activated by differentmechanisms. Activation of the PAR family of GPCRs has been associated with inflammation and nociception (Gieseler et al Cell Commun Signal. 2013; 11: 86).

[0006] FourPARreceptorshavebeen identified:PAR1,PAR2,PAR3,andPAR4(Macfarlaneetal.PharmacolRev. 2001 Jun;53(2):245‑82; Gieseler et al Cell Commun Signal. 2013; 11: 86). PAR2 activation has been shown to amplify inflammation and nociception, making its inhibition an attractive target for anti-inflammatory pain therapies. PARs, unlike other GPCRs, are activated by proteolytic cleavage of their extracellular domains, which reveals an N-terminal sequence that acts as a tethered-activating ligand. PAR2, in particular, is cleaved and activated by trypsin and tryptase.

[0007] PAR2 expression has been detected in vascularized tissues, airways, osteoblasts, cardiovascular tissue, keratinocytes, exocrine glands, leukocytes, mast cells, intestinal epithelium, kidney, neurons, pancreas, and a variety of smooth muscle types (Macfarlane supra). PAR2 has also been implicated in a variety of diseases or conditions associated with neurogenic inflammation, nociception and transmission of pain. PAR2 may be activated by several host and pathogen-derived serine proteases (e.g., trypsin,mast cell tryptase, tissue kallikreins, ormembers of the coagulation cascade TF-FVIIa and FVa-FXa).

[0008] Monoclonal antibodies have been shown to be useful in a variety of therapeutic applications andmany antibody 2 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 therapeutics are currently on the market (Maggon, Curr Med Chem. 2007;14(18):1978‑87; Brekke and Sandlie, Nat Rev Drug Discov 2: 52‑62, 2003). The most common type of antibody in circulation in the blood stream is immunoglobulin G (IgG). The usefulness of an IgG antibody for therapeutic purposes depends on several factors, including the specificity of the antibody for its target, the strength of its binding to the target, as well as how efficiently the antibody can be produced and how quickly the antibody is cleared from the serum (the serum half-life of the antibody). The serum half-lives of antibodies are frequently regulated by FcRn (neonatal Fc receptor), which binds to the Fc domain of immunoglobulin G (IgG). In vivo, IgGsare thought to be takenupnon-specifically by fluid-phasepinocytosis (Pyzik et al, J Immunol. 2015May 15;194(10):4595‑603).Once in theendosome, IgGbinds toFcRn,which sorts the IgG into recycling endosomesandback to the cell surface, away from lysosomesandaway fromdegradation.While the recycling rate of IgGhasbeenestimated to be 44% of the fractional catabolic rate, antibody therapeutics can still be depleted in a matter of days post-administration (Kim, Jonghan, et al. Clin. Immunol. 122.2 (2007): 146‑155).

[0009] Pain associated with inflammation is often a chronic condition. Minimizing the dosage and the frequency of administration of a therapeutic molecule is desirable. Thus there is a need for new anti-inflammation pain therapeutics. Standard monoclonal antibodies are attractive candidates, but in some cases can be limited by their serum half-lives. As such, alternative treatments may be desired. BRIEF SUMMARY OF THE DISCLOSURE

[0010] Thepresent disclosure providesantibodies that bindPAR2.Theantibodies of the disclosure areuseful, inter alia, for inhibitingPAR2-mediated signalingand for treatingdiseasesanddisorders causedbyor related toPAR2activity and / or signaling.

[0011] In some embodiments, the disclosure provides for antibodies or antigen-binding fragments thereof that bind to PAR2with agreater affinity at pH7.4 thanat pH6.0. In someembodiments, theantibodyor antigen-binding fragment binds to PAR2 with at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 times greater affinity at pH 7.4 than at pH 6.0. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ IDNO: 3, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 3; ii) a VH-CDR2 having the amino acid sequence of SEQ IDNO: 4, but wherein 1, 2, 3, 4, or 5 aminoacid substitutions, deletions or insertions are optionally present in the sequence ofSEQ IDNO: 4; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 5; and wherein the VL comprises: i) a VL- CDR1 having the amino acid sequence of SEQ IDNO: 8; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 8; ii) a VL-CDR2 having the amino acid sequence of SEQ IDNO: 9, butwherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 9; and iii) a VL-CDR3 having the amino acid sequence of SEQ IDNO: 10; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequenceofSEQ IDNO: 10;wherein theaminoacid substitutions, deletions or insertions reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having aVHwith an amino acid sequence of SEQ IDNO: 2 andVLwith an amino acid sequence of SEQ IDNO: 7when tested at a pH of 7.4 in a PAR2 binding assay. In some embodiments, the amino acid substitutions, deletions or insertions comprise a homologous substitution. In some embodiments, the antibody or antigen-binding fragment has nomore than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, insertions or deletions in the VHCDRs as compared to the CDR amino acid sequences present in the sequence of SEQ ID NO: 2. In some embodiments, the antibody or antigen-binding fragment has nomore than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, insertions or deletions in the VL CDRs as compared to the CDR amino acid sequences present in the sequence of SEQ ID NO: 7. In someembodiments, the 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10aminoacid substitutions, insertionsor deletions is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions with a histidine.

[0012] In some embodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) anda light chain variable domain (VL), wherein theVHcomprises: i) aVH-CDR1having the amino acid sequence of SEQ IDNO: 3; ii) a VH-CDR2having the amino acid sequence of SEQ IDNO: 4, but wherein a histidine isoptionallypresentat anyoneormoreof theaminoacidpositionscorresponding topositions1‑17 (e.g.,positions 4, 5, and 7‑17) of SEQ ID NO: 4; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein a histidine is optionally present at any one ormore of the amino acid positions corresponding to positions 1‑8 of SEQ IDNO: 5; andwherein theVLcomprises: i) aVL-CDR1having theaminoacidsequenceofSEQ IDNO:8; ii) aVL-CDR2having the aminoacid sequenceof SEQ IDNO: 9; and iii) a VL-CDR3having theamino acid sequence ofSEQ IDNO: 10; butwherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑14 of SEQ ID NO:10. Insomeembodiments, theVHcomprises: i) aVH-CDR1having theaminoacidsequenceofSEQIDNO:3, ii) aVH- CDR2having theaminoacid sequenceofSEQ IDNO:4, iii) aVH-CDR3having theaminoacid sequenceofSEQ IDNO: 5, 3 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 and the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8, ii) a VL-CDR2 having the amino acid sequenceofSEQ IDNO: 9, iii) aVL-CDR3having theaminoacid sequenceofSEQ IDNO: 10. In someembodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 13, ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 14, iii) a VH-CDR3 having the amino acid sequenceofSEQ IDNO: 15, andwherein theVL comprises: i) aVL-CDR1having theaminoacid sequenceof SEQ IDNO: 18, ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 19, iii) a VL-CDR3 having the amino acid sequence of SEQIDNO:20. Insomeembodiments,ahistidine ispresentat theaminoacidpositionscorresponding topositions5,8,12, 16, and 17ofSEQ IDNO: 4; andwherein a histidine is present at the aminoacid positions corresponding to positions 2 and 3 of SEQ ID NO: 5. In some embodiments, a histidine is present at the amino acid position corresponding to position 5 of SEQ IDNO: 4. In some embodiments, a histidine is present at the amino acid position corresponding to position 7 of SEQ IDNO:4. In someembodiments, ahistidine is present at theaminoacid position corresponding to position8ofSEQ IDNO: 4. In someembodiments, a histidine is present at the amino acid position corresponding to position 12 of SEQ IDNO: 4. In someembodiments, ahistidine ispresent at theaminoacidposition corresponding toposition15ofSEQ IDNO:4. In some embodiments, a histidine is present at the amino acid position corresponding to position 16 of SEQ ID NO: 4. In some embodiments, a histidine is present at the amino acid position corresponding to position 17 of SEQ ID NO: 4. In some embodiments, a histidine is present at the amino acid positions corresponding to positions 5 and 8 of SEQ ID NO: 4. In someembodiments, a histidine is present at the aminoacid positions corresponding to positions 5, 8, 12, and16ofSEQ ID NO: 4. In some embodiments, a histidine is present at the amino acid positions corresponding to positions 5, 8, 12, 16 and 17 of SEQ IDNO: 4. In some embodiments, a histidine is present at the amino acid position corresponding to position 2 of SEQ IDNO: 5. In some embodiments, a histidine is present at the amino acid position corresponding to position 3 of SEQ ID NO: 5. In some embodiments, a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ IDNO: 5. In some embodiments, a histidine is present at the amino acid position corresponding to position 1 of SEQ ID NO: 10. In some embodiments, a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 10. In some embodiments, a histidine is present at the amino acid position corresponding to position 6 of SEQ IDNO: 10. In someembodiments, a histidine is present at the amino acid position corresponding to position 14 of SEQ IDNO: 10. In some embodiments, the VH-CDR2 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794, and 811‑818. In some embodiments, the VH-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ IDNOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, and 819‑820. In some embodiments, the VL- CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790 and800. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 14; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ IDNO: 15, andwherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ IDNO: 811; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 819, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 814; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 820, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding toSEQ IDNO: 816; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 818; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ IDNO: 10 or 20. In some embodiments, the VH comprises framework regions that are at least 90% identical to eachofSEQ IDNOs: 803‑806. In someembodiments, theVHcomprises framework regions that are at least 95% identical to each of SEQ ID NOs: 803‑806. In some embodiments, the VH comprises framework regions corresponding toSEQ IDNOs: 803‑806. In someembodiments, theVLcomprises framework regions that areat least 90% identical to each of SEQ ID NOs: 807‑810. In some embodiments, the VL comprises framework regions that are at least 95% identical to each of SEQ IDNOs: 807‑810. In some embodiments, the VL comprises framework regions correspond- ing to SEQ IDNOs: 807‑810. In someembodiments, theVH comprises an amino acid sequence that is at least 80%, 85%, 4 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 90%, 92%, 93%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from the group consisting of: SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, and 792. In some embodiments, the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from the group consisting of: SEQ IDNOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797. In some embodiments, the VH comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ IDNO: 12 and wherein the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 17. In some embodiments, theVHcomprises anaminoacid sequencecorresponding toSEQ IDNO: 12andwherein theVLcomprises an amino acid sequence corresponding to SEQ ID NO: 17. In some embodiments, the VH comprises an amino acid sequence corresponding toSEQ IDNO:821and theVLcomprisesanaminoacid sequencecorresponding toSEQ IDNO: 7 or 17. In some embodiments, the VH comprises an amino acid sequence corresponding to SEQ ID NO: 824 and the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. In some embodiments, the VH comprises an amino acid sequence corresponding to SEQ IDNO: 827 and the VL comprises an amino acid sequence corresponding to SEQ IDNO: 7 or 17. In someembodiments, theVHcomprises an amino acid sequence corresponding to SEQ IDNO: 831 and theVL comprises an amino acid sequence corresponding to SEQ IDNO: 7 or 17. In someembodiments, the antibody or antigen-binding fragment is an antigen-binding fragment. In some embodiments, the antigen-binding fragment is an scFv. In some embodiments, the antigen-binding fragment is a Fab’. In some embodiments, the antibody or antigen- binding fragment is an antibody. In someembodiments, the antibody is amonoclonal antibody. In someembodiments, the antibody is an IgG antibody. In some embodiments, the antibody or antigen-binding fragment is humanized. In some embodiments, the antibody or antigen-binding fragment is human. In some embodiments, the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361,371, 381, 391, 401, 411,421, 431, 441, 451,461, 471, 481, 491, 501, 511,521, 531, 541,551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and833‑841. In some embodiments, the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251,261, 271, 281, 291, 301, 311,321, 331, 341,351, 361, 371, 381, 391, 401,411, 421, 431,441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. In some embodiments, the VH is encoded by a nucleic acid comprising thenucleotidesequenceof anyoneofSEQIDNOs:1, 11, 21,31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181,191, 201, 211,221, 231, 241, 251, 261, 271,281, 291, 301, 311,321, 331, 341, 351, 361,371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611,621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. In some embodiments, the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In someembodiments, theVL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In some embodiments, the VL is encoded by a nucleic acid comprising the nucleotide sequence of any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In some embodiments, theVH isencodedbyanucleic acid comprisinganucleotidesequence that is at least 90% identity toSEQID NO: 11. In some embodiments, the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity toSEQ IDNO: 11. In someembodiments, theVH is encodedbyanucleic acid comprising thenucleotide sequence of SEQ IDNO: 11. In some embodiments, the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 16. In some embodiments, the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 16. In some embodiments, the VL is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the antibody or antigen-binding fragment 5 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 binds to PAR2. In some embodiments, the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from interacting with PAR2. In some embodiments, the antibody or antigen-binding fragment prevents trypsin, tryptaseand / ormatriptase fromcleavingPAR2. In someembodiments, the antibodyor antigen-binding fragment prevents cleavage of the PAR2 extracellular domain. In some embodiments, the antibody or antigen-binding fragment inhibits exposure of the tethered ligand. In some embodiments, the antibody or antigen-binding fragment prevents the tethered ligand from interacting with the second transmembrane loop of PAR2. In some embodiments, the antibody or antigen- binding fragment binds toPAR2withgreater affinity at apHof 7.4 thanat apHof 6.0. In someembodiments, theantibodyor antigen-binding fragment has an IC50 of less than 100 nMwhen competing with an antibody or antigen-binding fragment having theCDRsof SEQ IDNOs: 3‑5 and 8‑10 at a pHof 7.4 in aPAR2binding assay. In someembodiments, the antibody or antigen-binding fragment has an IC50 of less than 50 nMwhen competingwith an antibody or antigen-binding fragment having theCDRsof SEQ IDNOs: 3‑5 and 8‑10 at a pHof 7.4 in aPAR2binding assay. In someembodiments, the antibody or antigen-binding fragment has an IC50 of less than 40 nMwhen competingwith an antibody or antigen-binding fragment having theCDRsof SEQ IDNOs: 3‑5 and 8‑10 at a pHof 7.4 in aPAR2binding assay. In someembodiments, the antibody or antigen-binding fragment has an IC50 of greater than 500 nM when competing with an antibody or antigen-binding fragment having theCDRsofSEQ IDNOs: 3‑5and8‑10at a pHof 6.0 in aPAR2binding assay. In someembodiments, the antibody or antigen-binding fragment has an IC50 of greater than 1000 nM when competing with an antibody or antigen- binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 6.0 in a PAR2 binding assay. In some embodiments, the antibody or antigen-binding fragment has an IC50 of greater than 1100 nM when competing with an antibodyor antigen-binding fragmenthaving theCDRsofSEQ IDNOs:3‑5and8‑10at apHof6.0 inaPAR2bindingassay. In some embodiments, the antibody or antigen-binding fragment has an IC50more than 20 times lower at a pH of 7.4 than at apHof 6.0whencompetingwithanantibodyor antigen-binding fragment having theCDRsofSEQ IDNOs: 3‑5and8‑10 in aPAR2binding assay. In someembodiments, the antibody or antigen-binding fragment has an IC50more than25 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ IDNOs: 3‑5 and 8‑10 in a PAR2 binding assay. In some embodiments, the antibody or antigen-binding fragment has an IC50 more than 30 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ IDNOs: 3‑5 and 8‑10 in a PAR2 binding assay. In some embodiments, the antibody or antigen-binding fragment has an IC50 of less than 3.0 x 10‑10 M in a calcium influx assay in human A549 cells. In some embodiments, the antibody or antigen-binding fragment has an IC50 of less than 1.5 x 10‑10 M in a calcium influx assay in humanA549 cells. In someembodiments, the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10‑10M in a calcium influx assay in rat KNRKcells. In someembodiments, the antibody or antigen-binding fragment has an IC50of less than 5.5 x 10‑10 M in a calcium influx assay in rat KNRK cells. In some embodiments, the antibody or antigen-binding fragment has an IC50of less than 7.0 x 10‑11M in a calcium influx assay in cynomolgusmonkeyCYNOM-K1 cells. In some embodiments, the antibody or antigen-binding fragment has an IC50 of less than 5.0 x 10‑11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells. In some embodiments, the antibody or antigen-binding fragment has an IC50 of less than6.0x10‑11M inacalcium influxassay inmurineLL / 2 cells. In someembodiments, theantibodyor antigen-binding fragment has an IC50 of less than 4.0 x 10‑11 M in a calcium influx assay in murine LL / 2 cells. In some embodiments, the antibody or antigen-binding fragment is cleared more slowly from serum of a treated patient than an antibody or antigen- binding fragment lacking histidine modifications. In some embodiments, the optionally present histidine or histidines reduce thebindingaffinity of the antibodyor antigen-binding fragment thereof for humanPAR2bynomore than1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having a VH with an aminoacid sequenceofSEQ IDNO: 2andVLwith anaminoacid sequenceofSEQ IDNO: 7when tested at a pHof 7.4 in a PAR2 binding assay.

[0013] In some embodiments, the disclosure provides for a nucleic acid capable of expressing any of the antibodies or antigen-binding fragments disclosed herein. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 90% identical to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191,201, 211,221, 231, 241, 251, 261, 271, 281,291, 301, 311,321, 331, 341, 351, 361, 371,381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511,521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and833‑841. In someembodiments, the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371,381, 391, 401, 411,421, 431, 441, 451, 461,471, 481, 491, 501, 511,521, 531, 541, 551,561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. In some embodiments, the nucleic acid comprises the nucleotide sequence of any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351,361, 371, 381, 391, 401, 411,421, 431, 441,451, 461, 471, 481, 491, 501,511, 521, 531,541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. In some embodiments, the disclosure provides for a nucleic acid comprising a nucleotide sequence that is at least 90% identical to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 6 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In some embodiments, the nucleic acid comprises thenucleotide sequence of any oneofSEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In some embodiments, the disclosure provides for a nucleic acid comprising a nucleotide sequence that is at least 90% identical to SEQ IDNO: 11. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ IDNO: 11. In someembodiments, the nucleic acid comprises the nucleotide sequence of SEQ IDNO: 11. In some embodiments, the disclosure provides for a nucleic acid comprising a nucleotide sequence that is at least 90% identical to SEQ IDNO: 16. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ IDNO: 16. In someembodiments, the nucleic acid comprises the nucleotide sequence of SEQ IDNO: 16.

[0014] In some embodiments, the disclosure provides for a vector comprising any of the the nucleic acids disclosed herein. In some embodiments, the disclosure provides for a set of vectors comprising any one ormore of the nucleic acids disclosed herein.

[0015] In someembodiments, thedisclosureprovides for ahost cell comprisinganyoneormoreof thevectors disclosed herein.

[0016] In some embodiments, the disclosure provides for a composition comprising a pharmaceutically acceptable carrier and any of the antibodies or antigen-binding fragments disclosed herein.

[0017] In some embodiments, the disclosure provides for a lyophilized composition comprising any of the antibody or antigen-binding fragment thereof disclosed herein.

[0018] In some embodiments, the disclosure provides for a reconstituted lyophilized composition comprising any of the antibodiesorantigen-binding fragments thereof disclosedherein. In someembodiments, thecomposition is formulated for administration by lozenge, spray, oral administration, delayed release or sustained release, transmucosal administration, syrup, mucoadhesive, buccal formulation, mucoadhesive tablet, topical administration, parenteral administration, injec- tion, subdermal administration, oral solution, rectal administration, buccal administration or transdermal administration.

[0019] In some embodiments, the disclosure provides for a kit comprising any of the antibodies or antigen-binding fragments disclosed herein or any of the compositions disclosed herein.

[0020] In some embodiments, the disclosure provides for a method for treating pain in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of any of the antibodies or antigen-binding fragments disclosed herein. In some embodiments, the disclosure provides for a method for treating pain in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of any of the compositions disclosed herein. In some embodiments, the pain is selected from the group consisting of: nociceptive, neuropathic, and mix-type pain. In some embodiments, the pain is associated with a headache, chronic headache, amigraine headache, a cancer, a viral infection, rheumatoid arthritis, osteoarthritis, Crohn’s disease, liver disease, multiple sclerosis, spinal cord injury, post herpetic neuralgia, diabetic neuropathy, lowerbackpain, inflammatory heart disease, kidneydisease, gastritis, gingivitis, periodontal disease, asthma, chronic obstructive pulmonary disease, autoimmune disease, irritable bowel syndrome, fibromyalgia, legpains, restless leg syndrome, diabetic neuropathy, anallergic condition, a surgical procedure, acute or chronic physical injury, bone fracture or a crush injury, spinal cord injury, an inflammatory disease, a non- inflammatory neuropathic or dysfunctional pain condition, or a combination thereof. In some embodiments, the pain is osteoarthritis pain. In some embodiments, the subject is a human.

[0021] In someembodiments, thedisclosureprovides foramethodofproducinganyof theantibodiesorantigen-binding fragments disclosed herein, comprising the steps of: expressing any of the the nucleic acids disclosed herein in a cultured cell, purifying the antibody or antigen-binding fragment. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] This patent or patent application filed contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. Figures 1A and1B are tables illustrating the sequence differences inVHVHCDR2 (SEQ IDNOS4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 7 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 314,324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, and 794, respectively, in order of appearance) andCDR3 (SEQ IDNOS5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, and795, respectively, in order of appearance) of various clonesas compared to the sameCDRsofPar0067.CDR1and the framework regions are the same for Par0067 and for all of the clones indicated (i.e., Par0067 and each of the clones comprised the sequences of SEQ ID NOs: 3 and 803‑806). Figures2Aand2Bare tables illustrating thesequencedifferences inVLCDR3 (SEQ IDNOS10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, and 800, respectively, in order of appearance) of various clones as compared to the same CDR of Par0067. CDR1, CDR2 and the framework regions are the same for Par0067 and for all of the clones indicated (i.e., the clones comprised the sequences of SEQ ID NOs: 8, 9, and 807‑810). Figure 3 provides IC50 data from a cell potency assay using various IgG-based antibodies. The types of cells used in each of the cell assays are indicated. NI = non-inhibitory. Figure 4A provides IC50 curves for PaB670129 against trypsin in A549 cells, relative to agonistic responses to the antibody at the equivalent concentrations. Figure 4B provides IC50 curves for PaB670129 against different PAR2 protease activators. Figures 5A‑5F illustrate the results from experiments in which rat dorsal root ganglia (DRG) sensory neurons or non- neuronal cells were treated with matripase in the presence or absence of PaB670129 (also referred to herein as PaB670129). Rat DRG sensory neurones pre-treated with isotype control (20 nM) display matriptase-induced calcium transients (5A). Sensory neurons pre-treatedwith PaB670129 IgG1TM (20 nM) do not respond tomatriptase (5B); % of neurones responding to matriptase quantified in (5C). Non-neuronal cells of the DRG pre-treated with isotype control (20 nM) also display matriptase-induced calcium transients (8D), but non-neuronal cells pre-treated with PaB670129 IgG1TM (20 nM) do not (5E); % of non-neuronal cell responding to matriptase quantified in (5F). Figures 5A‑5E disclose "LIGRLO" as SEQ ID NO: 832 Figure 6 illustrates the effects of PaB670129 (versus anti-PAR1 antibodies or Vorapaxar) on thrombin-induced PAR1 activation in human A549 cells. Figure 7A depicts a graph illustrating the effect of different treatments (including a PAR2 antibody, Par0067) on the percent ipsilateral / contralateral hypersensitivity induced bymono-iodoacetate (MIA) in rat. Figure 7B depicts a graph illustrating the effect of different doses of PaB670129 or an isotype control antibody on the percent ipsilateral / con- tralateral hypersensitivity induced by MIA in rat. "i.v." means intravenous, and "p.o." means per os (oral). Figure 8 depicts a graph illustrating the effect of different doses of PaB670129 or an isotype control antibody on the percent ipsilateral / contralateral hypersensitivity induced by peripheral nerve partial ligation in mouse. "s.c." means sub-cutaneous. N= 9‑10 per group. Data were analyzed using 2-way ANOVAwith time and treatment as dependent factors. Subsequent statistical significancewas obtained using Tukey’sPost Hoc test. Individual comparisons shown * P<0.05; ** P<0.01; *** P<0.001 vs. Op + isotype control 10 mg / kg. DETAILED DESCRIPTION

[0023] Before the present disclosure is described, it is to be understood that this disclosure is not limited to particular methodsandexperimental conditionsdescribed, as suchmethodsandconditionsmayvary. It is also tobeunderstood that the terminology usedherein is for the purposeof describing particular embodiments only, and is not intended to be limiting.

[0024] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0025] Althoughanymethodsandmaterials similar or equivalent to thosedescribedherein canbeused in thepracticeor testing of the present disclosure, the preferred methods and materials are now described. A. Definitions

[0026] Asused in this specificationand theappendedclaims, thesingular form"a", "an"and "the" includeplural referents unless the context clearly dictates otherwise.

[0027] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes. 8 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55

[0028] It is convenient to point out here that "and / or"whereusedherein is tobe takenasspecific disclosureof eachof the two specified features or components with or without the other. For example "A and / or B" is to be taken as specific disclosure of each of (i) A, (ii) B and (iii) A and B, just as if each is set out individually herein.

[0029] The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, aswell as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymer.

[0030] The expressions "protease activated receptor 2," "PAR2," and the like, as used herein, refer to a human PAR2 protein having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any of SEQ ID NO: 801, or biologically active fragments thereof.

[0031] The term "tethered ligand" refers to a region of the N-terminal portion of PAR2 that binds to and activates the PAR2 receptor itself. In some embodiments, the tethered ligand portion of PAR2 is not exposed until a protease (e.g., thrombin or trypsin) proteolytically cleaves a portion of the PAR2 enzyme. In some embodiments, the tethered ligand corresponds to a polypeptide that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any of SEQ ID NO: 802

[0032] As used herein, "an antibody that binds PAR2", "anti-PAR2 antibody," and the like includes antibodies, and antigen-binding fragments thereof, that bind membrane-bound PAR2 or fragments thereof. In some embodiments, an anti-PAR2 antibody or antigen binding fragment thereof binds to the tethered ligand portion of PAR2. B. Antibodies and Antigen-Binding Fragments Thereof

[0033] As used herein, "antibodies or antigen binding fragments of the disclosure" refer to any one or more of the antibodies and antigen binding fragments provided herein. Antibodies and antigen binding fragments of the disclosure comprise a heavy chain (VH) comprising a heavy chain variable domain and a light chain (VL) comprising a light chain variable domain. A VH domain comprises three CDRs, such as any of the CDRs provided herein and as defined or identified by the Chothia, Kabat or IMGTsystems. These CDRs are typically interspersed with framework regions (FR), and together comprise the VH domain. Similarly, a VL comprises three CDRs, such as any of the CDRs provided herein and as defined by the Chothia, Kabat or IMGTsystems. These CDRs are typically interspersed with framework regions (FR), and together comprise the VL domain. The FR regions, such as FR1, FR2, FR3, and / or FR4 can similarly be defined or identified by the Chothia, Kabat or IMGTsystems. Throughout the application, when CDRs are indicated as being, as identified or as defined by theChothia, Kabat or IMGTsystems, what ismeant is that theCDRs are in accordancewith that system (e.g., theChothia CDRs, Kabat CDRs or the IMGTCDRs). Any of these terms can be used to indicate whether the Chothia, Kabat or IMGT CDRs are being referred to.

[0034] The term "antibody", as used herein, also includes antigen-binding fragments of full antibody molecules. The terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like, as used herein, includeanynaturally occurring, enzymatically obtainable, synthetic, or genetically engineeredpolypeptide or glycoprotein that specifically binds an antigen to form a complex. Antigen-binding fragments of an antibodymay be derived, e.g., from full antibody molecules using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable and optionally constant domains.SuchDNA is knownand / or is readily available from,e.g., commercial sources,DNA libraries (including, e.g., phage-antibody libraries), or can be synthesized. The DNA may be sequenced and manipulated chemically or by usingmolecular biology techniques, for example, to arrange one ormore variable and / or constant domains into a suitable configuration, or to introduce codons, create cysteine residues, modify, add or delete amino acids, etc.

[0035] In some embodiments, the disclosure provides for antibodies or antigen-binding fragments thereof that bind to PAR2with agreater affinity at pH7.4 thanat pH6.0. In someembodiments, theantibodyor antigen-binding fragment binds to PAR2 with at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 times greater affinity at pH 7.4 than at pH 6.0. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ IDNO: 3, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 3; ii) a VH-CDR2 having the amino acid sequence of SEQ IDNO: 4, but wherein 1, 2, 3, 4, or 5 aminoacid substitutions, deletions or insertions are optionally present in the sequence ofSEQ IDNO: 4; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 5; and wherein the VL comprises: i) a VL- CDR1 having the amino acid sequence of SEQ IDNO: 8; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 8; ii) a VL-CDR2 having the amino acid sequence of SEQ IDNO: 9, butwherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 9; and iii) a VL-CDR3 having the amino acid sequence of SEQ IDNO: 10; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequenceofSEQ IDNO: 10;wherein theaminoacid 9 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 substitutions, deletions or insertions reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50, 10 or 5-fold as compared to an antibody or antigen-binding fragment having aVHwith an amino acid sequence of SEQ IDNO: 2 andVLwith an amino acid sequence of SEQ IDNO: 7when tested at a pH of 7.4 in a PAR2 binding assay. In some embodiments, the amino acid substitutions, deletions or insertions comprise a homologous substitution. In some embodiments, the antibody or antigen-binding fragment has nomore than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, insertions or deletions in the VHCDRs as compared to the CDR amino acid sequences present in the sequence of SEQ ID NO: 2. In some embodiments, the antibody or antigen-binding fragment has nomore than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, insertions or deletions in the VL CDRs as compared to the CDR amino acid sequences present in the sequence of SEQ ID NO: 7. In someembodiments, the 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10aminoacid substitutions, insertionsor deletions is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions with a histidine.

[0036] In some embodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) anda light chain variable domain (VL), wherein theVHcomprises: i) aVH-CDR1having the amino acid sequence of SEQ IDNO: 13, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionallypresent in thesequenceofSEQIDNO:13; ii) aVH-CDR2having theaminoacidsequenceofSEQIDNO:14,but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 14; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 15, but wherein 1, 2, 3, 4, or 15 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 15; and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 18; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 18; ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 19, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 19; and iii) a VL-CDR3 having the amino acid sequence of SEQ IDNO: 20; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ IDNO: 20; wherein the amino acid substitutions, deletions or insertions reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50, 10 or 5- fold as compared to an antibody or antigen-binding fragment having a VHwith an amino acid sequence of SEQ IDNO: 12 and VL with an amino acid sequence of SEQ ID NO: 17 when tested at a pH of 7.4 in a PAR2 binding assay. In some embodiments, the amino acid substitutions, deletions or insertions comprise a homologous substitution. In some embodiments, the antibody or antigen-binding fragment has no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, insertions or deletions in the VH CDRs as compared to the CDR amino acid sequences present in the sequence of SEQ IDNO: 12. In some embodiments, the antibody or antigen-binding fragment has nomore than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, insertions or deletions in the VL CDRs as compared to the CDR amino acid sequences present in the sequence of SEQ ID NO: 17. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 13, ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 14, iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 15, and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ IDNO: 18, ii) a VL-CDR2 having the amino acid sequenceofSEQ IDNO:19, iii) aVL-CDR3having theaminoacid sequenceofSEQ IDNO:20. In someembodiments, the 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10aminoacid substitutions, insertionsor deletions is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutionswith a histidine.

[0037] In someembodiments, thedisclosureprovides for anantibodyor antigen-binding fragment,wherein theantibody orantigen-binding fragment comprisesa light chainvariable (VL)domainandaheavychainvariable (VH)domain;wherein the VH domain comprises at least one, two or all three of the CDRs (e.g., as measured using any of the Chothia, IMGTor Kabat systems) of the amino acid sequence set forth in any one of SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112,122,132, 142,152,162, 172,182,192, 202,212,222, 232,242,252, 262,272, 282,292,302, 312,322,332, 342,352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, and 792. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment, wherein the antibody or antigen- binding fragment comprises a light chain variable (VL) domain and a heavy chain variable (VH) domain; wherein the VH domain comprises CDR1, CDR2 and CDR3 (e.g., as measured using any of the Chothia, IMGTor Kabat systems) of the amino acid sequence set forth in any oneof SEQ IDNOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, and 792. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment, wherein the antibody or antigen-binding fragment comprises a light chain variable (VL) domainandaheavychain variable (VH) domain;wherein theVHdomain comprisesat least one, twoor all three of theCDRs (e.g., asmeasured using any of theChothia, IMGTorKabat systems) of the amino acid sequence set forth in SEQ ID NO: 2 or 12. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment, 10 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 wherein the antibody or antigen-binding fragment comprises a light chain variable (VL) domain anda heavy chain variable (VH) domain; wherein the VL domain comprises at least one, two or all three of the CDRs (e.g., asmeasured using any of theChothia, IMGTor Kabat systems) of the amino acid sequence set forth in any one of SEQ IDNOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and797. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment, wherein the antibody or antigen-binding fragment comprises a light chain variable (VL) domain and a heavy chain variable (VH) domain; wherein theVLdomaincomprisesCDR1,CDR2andCDR3(e.g.,asmeasuredusinganyof theChothia, IMGTorKabat systems)of the amino acid sequence set forth in any one of SEQ IDNOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment, wherein the antibody or antigen-binding fragment comprisesa light chain variable (VL)domainandaheavychainvariable (VH)domain;wherein theVLdomaincomprisesat least one, two or all three of the CDRs (e.g., asmeasured using any of the Chothia, IMGTor Kabat systems) of the amino acid sequence set forth in SEQ IDNO: 7 or 17. In some embodiments, the disclosure provides for an antibody or antigen- binding fragment, wherein the antibody or antigen-binding fragment comprises a light chain variable (VL) domain and a heavy chain variable (VH) domain; wherein the VH domain comprises CDR1, CDR2 and CDR3 (e.g., asmeasured using any of the Chothia, IMGTor Kabat systems) of the amino acid sequence set forth in any one of SEQ IDNOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, and792,andwherein theVLdomaincomprisesCDR1,CDR2andCDR3 (e.g.,asmeasuredusinganyof theChothia, IMGTorKabat systems)of theaminoacid sequenceset forth inanyoneofSEQ IDNOs:7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107,117,127, 137,147,157, 167,177,187, 197,207,217, 227,237,247, 257,267, 277,287,297, 307,317,327, 337,347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797. In some embodiments, the disclosure provides for an antibody or antigen-binding fragment, wherein the antibody or antigen- binding fragment comprises a light chain variable (VL) domain and a heavy chain variable (VH) domain; wherein the VH domain comprises CDR1, CDR2 and CDR3 (e.g., as measured using any of the Chothia, IMGTor Kabat systems) of the aminoacidsequenceset forth inSEQIDNO:2or 12,andwherein theVLdomaincomprisesCDR1,CDR2andCDR3(e.g., asmeasuredusinganyof theChothia, IMGTorKabat systems)of theaminoacid sequenceset forth inSEQ IDNO:7or 17.

[0038] Once the nucleotide sequences encoding such antibodies have been determined, chimeric or humanized antibodiesmaybeproducedby recombinantmethods.Nucleic acidsencoding theantibodiesare introduced intohost cells andexpressedusingmaterialsandproceduresgenerally known in theart, andasdisclosedherein. In someembodiments, the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%or 100% identity to any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151,161, 171, 181, 191, 201, 211,221, 231, 241,251, 261, 271, 281, 291, 301,311, 321, 331,341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. In some embodiments, theVH isencodedbyanucleic acid comprisinganucleotide sequence that is at least 80%,85%,90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 1 or 11. In some embodiments, the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. In some embodiments, the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 6 or 16.

[0039] The present disclosure includes anti-PAR2 antibodies and antigen-binding fragments thereof that bind PAR2. In some embodiments, the antibody is a neutralizing and / or blocking anti-PAR2 antibody or antigen-binding fragment. A "neutralizing" or "blocking" antibody or antigen-binding fragment, as used herein, is intended to refer to an antibody or antigen-binding fragment whose binding to PAR2: (i) interferes with the interaction between PAR2 and a protease (e.g., trypsin, tryptase and / or matriptase); (ii) inhibits the cleavage of PAR2 by a protease; (iii) inhibits PAR2 signalling or PAR2 activation; and / or (iv) results in inhibition of at least one biological function of PAR2. In some embodiments, the antibodies or antigen-binding fragments of the disclosure inhibit activation of PAR2. In someembodiments, the antibodies or antigen- binding fragments inhibit conversion of inactive, uncleaved PAR2 into active, cleaved PAR2. In some embodiments, the 11 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 antibodies or antigen-binding fragments inhibit exposure of the tethered ligand. In some embodiments, the antibodies or antigen-binding fragments inhibit activation of a PAR2 receptor by its tethered ligand. In some embodiments, the antibodies or antigen-binding fragments inhibit binding of the tethered ligand to the second transmembrane domain of PAR2. The inhibition caused by an anti-PAR2 neutralizing or blocking antibody need not be complete so long as it is detectable using an appropriate assay. In some embodiments, the antibody or antigen-binding fragment thereof inhibits PAR2activity at least 10%,20%, 30%, 40%, 50%, 60%,70%, 80%, 90%or 100%ascompared touninhibited activePAR2. Some examples of assays for detecting activity of a representative anti-PAR2 antibody or antigen-binding fragment are described in the Exemplification section. The skilled worker is aware of additional anti-PAR2 antibody activity assays.

[0040] In particular embodiments, any of the antibodies or antigen-binding fragments disclosed herein interferes with the interaction between PAR2 and a protease. In some embodiments, the protease is trypsin. In some embodiments, the protease is neutrophil elastase. In someembodiments, the protease is neutrophil proteinase3. In someembodiments, the protease is mast cell tryptase. In some embodiments, the protease is tissue factor / factor VIIa / factor Xa. In some embodiments, the protease is a kallikrein-related peptidase. In some embodiments, the protease is membrane-tethered serine proteinase‑1 / matriptase 1. In some embodiments, the protease is parasite cysteine proteinase. In some embodi- ments, the anti-PAR2 antibodies or antigen-binding fragments block the interaction between PAR2 and a protease (e.g., trypsin) in vitro,with an IC50 value of less than about 15 nM, asmeasured by a binding assay such as that described in the Exemplification section. In certain embodiments, the antibodies or antigen-binding fragments of the present disclosure block the interaction betweenPAR2andaprotease (e.g., trypsin) in vitroat a pHof 7.4with an IC50 value of less than about 200nM,150nM,100nM,50nM,40nM,30nM,20nM,10nM,1nM,500pM,400pM,200pM,100pM,50pM,5pM,1pMor 0.1pM. Incertainembodiments, theantibodiesorantigen-binding fragmentsof thepresentdisclosureblock the interaction betweenPAR2and a protease (e.g., trypsin) in vitro at a pH of 6.0with an IC50 value of greater than about 300nM, 500 nM, 750 nM, 1000 nM, 1100 nM, or 1200 nM. In certain embodiments, the IC50 of the anti-PAR2antibody or fragment thereof is measured in anepitopecompetition assay, suchas theepitopecompetitionassaydescribed in theExemplification section provided herein. In some embodiments, the IC50 of the anti-PAR2 antibody or fragment thereof is measured in a cell potency assay. In some embodiments, the cell potency assay utilizes a human cell (e.g.,A549 cell), a rat cell (e.g.,KNRK cell), a cynomolgusmonkey cell (e.g.,CYNOM-K1 cell), or amurine cell (e.g.,an LL / 2 cell). In someembodiments, the cell potency assay utilizes a calcium influx assay (e.g., the calcium influx assay described in the Exemplification section provided herein). In some embodiments, the antibody or antigen-binding fragment inhibits calcium influx in the calcium influx assaywith an IC50 of less than 1nM, 500 pM, 400 pM, 200 pM, 100 pM, 50 pM, 10 pM, 5 pM, 1 pMor 0.1 pM. In some embodiments, the antibodies or antigen-binding fragments prevent abnormal activation of PAR2 by trypsin. In some embodiments, the antibodies or antigen-binding fragments inhibit / reduce inflammation-induced pain.

[0041] In particular embodiments, any of the antibodies or antigen-binding fragments disclosed herein interferewith the interaction between PAR2 and a protease (e.g., trypsin). In some embodiments, the antibodies prevent a protease (e.g., trypsin) from binding, cleaving, and / or activating PAR2. The present disclosure provides for anti-PAR2 antibodies and antigen-binding fragments thereof that bind PAR2 molecules with high affinity at physiological, extracellular pH (i.e. pH 7.4). In some embodiments, antibodies and antigen-binding fragments of antibodies bind PAR2 at pH 7.4 (e.g., at 25°C or 37°C)withaKDof less thanabout5nM,1nM,900pM,800pM,700pM,650pM,600pM,500pM,200pM,100pMor50pM. In some embodiments, antibodies and antigen-binding fragments of antibodies bind PAR2 at a slightly acidic pH (such as pH 6.0) (e.g., at 25°C or 37°C) with a KD of greater than about 1 nM, 5 nM, 10 nM, 15 nM, 20 nM, 25 nM, 30 nM, 40 nM, 50 nM, 60 nM, 80 nM, or 100 nM. In some embodiments, the slightly acidic pH is the pH of an endosomal compartment. In someembodiments,KDcanbemeasured inaccordancewith currently standardmethods, suchasusingSurfacePlasmon Resonance (SPR) or Quartz Crystal Microbalance (QCM).

[0042] The present disclosure also includes anti-PAR2 antibodies and antigen-binding fragments thereof that speci- fically bind to PAR2 with a dissociative half-life (t1 / 2) of greater than about 1.5 minutes, 1.75 minutes, 2 minutes, 2.5 minutes, 3 minutes, 5 minutes, 10 minutes, 20 minutes, or 30 minutes as measured using an assay such as surface plasmon resonance at 25°C or 37°C at pH 7.4. In some embodiments, anti-PAR2 antibodies and antigen-binding fragments thereof bind to PAR2 with a dissociative half-life (t1 / 2) of less than about 1 minute, 45 seconds, 30 seconds, 20 seconds, 15 seconds, 13 seconds, 7 seconds, 5 seconds, or 3 seconds as measured using an assay such as surface plasmon resonance at 25°C or 37°C at a slightly acidic pH (e.g., pH 6). In some embodiments, the slightly acidic pH is the pH of an endosomal compartment. In some embodiments, KD can be measured in accordance with currently standard methods, such as using Surface Plasmon Resonance (SPR) or Quartz Crystal Microbalance (QCM).

[0043] The antibodies or antigen-binding fragments of the present disclosure may possess one or more of the aforementioned biological characteristics, or any combinations thereof. Other biological characteristics of the antibodies of the present disclosure will be evident to a person of ordinary skill in the art from a review of the present disclosure including the Exemplification section provided herein.

[0044] As applied to polypeptides, the term "substantial similarity" or "substantially similar" means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least 95% sequence identity, even more preferably at least 98% or 99% sequence identity. Preferably, residue positions which 12 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 are not identical differ by conservative amino acid substitutions. In some embodiments, any of the antibodies or antigen- binding fragments disclosed herein comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 conservative aminoacid substitutions as compared to a reference sequence (e.g., any of the the aminoacid sequences of SEQ IDNOs: 2, 7, 12 or 17). A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative aminoacid substitutionwill not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degreeof similaritymaybeadjustedupwards to correct for the conservativenature of the substitution.Means for making thisadjustmentarewell-known to thoseof skill in theart.See,e.g.,Pearson (1994)MethodsMol.Biol. 24:307‑331. Examples of groups of amino acids that have side chainswith similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide- containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains are cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine- isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine.

[0045] Alternatively, a conservative replacement is any change having a positive value in the PAM250 log-likelihood matrix disclosed inGonnet et al. (1992)Science 256: 1443‑1445. A "moderately conservative" replacement is any change having a nonnegative value in the PAM250 log-likelihood matrix.

[0046] Depending on the amino acid sequences of the constant domains of their heavy chains, antibodies (immu- noglobulins) canbeassigned todifferent classes. Thereare fivemajor classesof immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are calledα, δ, ε, γ, andµ, respectively. Thesubunit structures and three-dimensional configurationsof different classesof immunoglobulins arewell known and described generally in, for example, Abbas et al. Cellular and Mol. Immunology, 4th ed. (W.B. Saunders, Co., 2000). Anantibodymaybepart of a larger fusionmolecule, formedby covalent or non-covalent association of the antibody with one or more other proteins or peptides.

[0047] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) Fab’ fragments; (iii) F(ab’)2 fragments; (iv) Fd fragments; (v) Fv fragments; (vi) single-chainFv (scFv)molecules; (vii) dAb fragments; and (viii)minimal recognition units consisting of the aminoacid residues thatmimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, cameliid antibodies, domain- deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nano- bodies (e.g. monovalent nanobodies, bivalent nanobodies, etc.), adnectins, small modular immunopharmaceuticals (SMIPs), and shark variable IgNARdomains, are also encompassedwithin the expression "antigen-binding fragment," as used herein.

[0048] An antigen-binding fragment of an antibodywill typically comprise at least one variable domain (e.g., at least one of a VHor VL). The variable domainmay be of any size or amino acid composition andwill generally comprise at least one CDR which is adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a VH domain associated with a VL domain, the VH and VL domains may be situated relative to one another in any suitable arrangement. For example, the variable regionmay bedimeric and contain VH-VH, VH-VLor VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.

[0049] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked toat least oneconstant domain.Non-limiting, exemplary configurationsof variableandconstant domains thatmaybe foundwithin an antigen-binding fragment of an antibody of the present disclosure include: (i) VH-CH1 ; (ii) VH- CH2; (iii) VH‑CH3; (iv) VH-CH1-CH2; (V) VH-CH1 ‑CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1 ; (ix) VL-CH2; (x)VL-CH3; (xi)VL-CH1-CH2; (xii)VL-CH1-CH2-CH3; (xiii) VL-CH2-CH3;and (xiv)VL-CL. Inanyconfigurationof variable and constant domains, including anyof theexemplary configurations listedabove, the variable and constant domainsmay be either directly linked to oneanother ormaybe linked by a full or partial hinge or linker region. A hinge regionmay consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids which result in a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. In some embodiments, the hinge region comprises a glycine-serine linker.

[0050] Moreover, an antigen-binding fragment of an antibody of the present disclosure may comprise a homo-dimer or hetero-dimer (or other multimer) of any of the variable and constant domain configurations listed above in non-covalent association with one another and / or with one or more monomeric VH or VL domain (e.g., by disulfide bond(s)).

[0051] As with full antibody molecules, antigen-binding fragments may be monospecific or multispecific (e.g., bispe- cific).Amultispecificantigen-binding fragment of anantibodywill typically compriseat least twodifferent variabledomains, wherein each variable domain is capable of specifically binding to a separate antigen or to a different epitope on the same antigen. Any multispecific antibody format, including the exemplary bispecific antibody formats disclosed herein, may be 13 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 adapted for use in the context of an antigen-binding fragment of an antibody of the present disclosure using routine techniques available in the art.

[0052] In certain embodiments of the disclosure, the anti-PAR2 antibodies of the disclosure are human antibodies. The term "human antibody", as used herein, is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site- specificmutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs, and in some embodiments, CDR3. However, the term "human antibody", as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0053] The antibodies of the disclosure may, in some embodiments, be recombinant human antibodies. The term "recombinant human antibody", as used herein, is intended to include all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant, combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287‑6295) or antibodies prepared, expressed, created or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies are subjected to in vitromutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somaticmutagenesis) and thus the amino acid sequences of theVHandVL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo.

[0054] Human antibodies can exist in two forms that are associated with hinge heterogeneity. In one form, an immunoglobulin molecule comprises a stable four chain construct of approximately 150‑160 kDa in which the dimers are held together by an interchain heavy chain disulfide bond. In a second form, the dimers are not linked via inter-chain disulfidebondsandamoleculeof about 75‑80kDa is formedcomposedof a covalently coupled light andheavy chain (half- antibody). These forms have been extremely difficult to separate, even after affinity purification.

[0055] The frequency of appearance of the second form in various intact IgG isotypes is due to, but not limited to, structural differencesassociatedwith the hinge region isotype of the antibody. A single aminoacid substitution in the hinge region of the human IgG4 hinge can significantly reduce the appearance of the second form (Angal et al. (1993)Molecular Immunology 30:105) to levels typically observed using a human IgG1 hinge. The current disclosure contemplates antibodies having one or more mutations in the hinge, CH2 or CH3 region which may be desirable, for example, in production, to improve the yield of the desired antibody form.

[0056] The antibodies of the disclosure may be isolated antibodies or isolated antigen-binding fragments. An "isolated antibody" or "isolated antigen-binding fragment," as used herein,means an antibody or antigen-binding fragment that has been identified and separated and / or recovered from at least one component of its natural environment. For example, an antibody or antigen-binding fragment that has been separated or removed fromat least one component of an organism, or from a tissue or cell in which the antibody naturally exists or is naturally produced, is an "isolated antibody" or an "isolated antigen-binding fragment" for purposes of the present disclosure. An isolated antibody also includes an antibody in situ within a recombinant cell. Isolated antibodies or antigen-binding fragments are antibodies or antigen-binding fragments that have been subjected to at least one purification or isolation step. According to certain embodiments, an isolated antibody or antigen-binding fragment may be substantially free of other cellular material and / or chemicals.

[0057] The anti-PAR2 antibodies or antigen-binding fragments disclosed herein may comprise one or more amino acid substitutions, insertions and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains as compared to the corresponding germline sequences from which the antibodies were derived. The present disclosure includes antibodies, and antigen-binding fragments thereof, which are derived from any of the amino acid sequences disclosed herein, wherein one or more amino acids within one or more framework and / or CDR regions are mutated to the corresponding residue(s) of the germline sequence from which the antibody or antigen-binding fragment was derived, or to the corresponding residue(s) of another human germline sequence, or to a conservative amino acid substitution of the corresponding germline residue(s) (such sequence changes are referred to herein collectively as "germlinemutations"). Apersonof ordinary skill in theart, startingwith theheavyand light chain variable regionsequences disclosed herein, can easily produce numerous antibodies and antigen-binding fragments which comprise one or more individual germlinemutations or combinations thereof. In certain embodiments, all of the framework and / or CDR residues within the VHand / or VL domains aremutated back to the residues found in the original germline sequence fromwhich the antibody was derived. In other embodiments, only certain residues are mutated back to the original germline sequence, e.g., only themutated residues foundwithin the first 8 aminoacidsof FR1orwithin the last 8 aminoacidsof FR4, or only the mutated residues found within CDR1, CDR2 or CDR3. In other embodiments, one or more of the framework and / or CDR 14 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 residue(s) aremutated to the corresponding residue(s) of a different germline sequence (i.e., a germline sequence that is different from the germline sequence from which the antibody was originally derived). In some embodiments, the VH framework region 1 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%or 100% identical to the sequence of SEQ IDNO: 803. In some embodiments, the VH framework region 2 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence of SEQ ID NO: 804. In some embodiments, the VH framework region 3 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%or 100% identical to thesequenceofSEQIDNO:805. Insomeembodiments, theVH framework region4comprisesanaminoacidsequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence of SEQ ID NO: 806. In some embodiments, the VL framework region 1 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%or 100% identical to the sequenceofSEQ IDNO: 807. In some embodiments, the VL framework region 2 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence of SEQ ID NO: 808. In some embodiments, the VL framework region 3 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence of SEQ IDNO: 809. In some embodiments, the VL framework region 4 comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence of SEQ ID NO: 810. In some embodiments, the VH framework comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 conservative substitutions as compared to a reference sequence of any one of SEQ ID NOs: 803‑806. In some embodiments, the VL framework comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 conservative substitutions as compared to a reference sequence of any one of SEQ ID NOs: 807‑810.

[0058] Furthermore, the antibodies of the present disclosure may contain any combination of two or more germline mutations within the framework and / or CDR regions, e.g., wherein certain individual residues are mutated to the corresponding residue of a particular germline sequencewhile certain other residues that differ from the original germline sequence are maintained or are mutated to the corresponding residue of a different germline sequence. Once obtained, antibodiesandantigen-binding fragments that containoneormoregermlinemutationscanbeeasily tested foroneormore desired property such as, improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as the case may be), reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained in this general manner are encompassed within the present disclosure.

[0059] Thepresent disclosure also includes anti-PAR2antibodies comprising variants of any of theVH,VL, and / or CDR amino acid sequences disclosed herein having one or more conservative substitutions. For example, the present disclosure includes anti-PAR2 antibodies having VH, VL, and / or CDR amino acid sequences with, e.g., 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 conservative amino acid substitutions relative to any of the VH, VL, and / or CDR amino acid sequences disclosed herein.

[0060] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen binding site in the variable region of an antibody molecule known as a paratope. A single antigen may have more than one epitope. Thus, different antibodies may bind to different areas on an antigen and may have different biological effects. Epitopes may be either conformational or linear. A conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain. A linear epitope is one produced by adjacent amino acid residues in a polypeptide chain. In certain circumstance, an epitope may include moieties of saccharides, phosphoryl groups, or sulfonyl groups on the antigen.

[0061] It should be noted that any portion of any of the antibodies or antigen-binding fragments of the disclosuremay be similarlymodified, suchaswith anepitope tag, aPEGmoietyormoieties, and the like.Moreover, theantibodiesor antigen- binding fragments may comprise more than one epitope tags, such as 2 epitope tags, or may include 0 epitope tags.

[0062] The term "substantial identity" or "substantially identical," when referring to a nucleic acid or fragment thereof, indicates that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 95%, and more preferably at least about 96%, 97%, 98% or 99% of the nucleotide bases, as measured by any well-known algorithm of sequence identity, such as FASTA, BLASTorGap, as discussed below. A nucleic acidmolecule having substantial identity to a reference nucleic acid moleculemay, in certain instances, encodeapolypeptide having the sameor substantially similar aminoacid sequenceas the polypeptide encoded by the reference nucleic acid molecule.

[0063] Sequence similarity for polypeptides, which is also referred to as sequence identity, is typically measured using sequence analysis software. Protein analysis softwarematches similar sequences usingmeasures of similarity assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, GCG software contains programs such as Gap and Bestfit which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild type protein and a mutein thereof. See, e.g., GCG Version 6.1 . Polypeptide sequences also can be compared using FASTA using default or recommended parameters, a program in GCGVersion6.1.FASTA (e.g., FASTA2andFASTA3)providesalignmentsandpercent sequence identity of the regionsof 15 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 the best overlap between the query and search sequences (Pearson (2000) supra). Another preferred algorithm when comparinga sequenceof the disclosure to adatabase containing a largenumber of sequences fromdifferent organisms is the computer programBLAST, especiallyBLASTPorTBLASTN,usingdefault parameters.See, e.g.,Altschul et al. (1990) J. Mol. Biol. 215:403‑410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389‑402. In some embodiments, the sequences are compared using EMBOSS Needle pairwise sequence alignment.

[0064] In some embodiments, the antibody or antigen-binding fragment is an antigen-binding fragment. In some embodiments, the antigen-binding fragment is an scFv. In some embodiments, the antigen-binding fragment is a Fab’. In some embodiments, the antibody or antigen-binding fragment is an antibody. In some embodiments, the antibody is a monoclonal antibody.

[0065] Antibodies became useful and of interest as pharmaceutical agents with the development of monoclonal antibodies. Monoclonal antibodies are produced using any method that produces antibody molecules by continuous cell lines in culture. Examples of suitablemethods for preparingmonoclonal antibodies include the hybridomamethods of Kohler et al. (1975, Nature 256:495‑497) and the human B-cell hybridomamethod (Kozbor, 1984, J. Immunol. 133:3001; andBrodeur et al., 1987,Monoclonal Antibody Production Techniques andApplications, (Marcel Dekker, Inc., NewYork), pp. 51‑63). In many cases, hybridomas are used to generate an initial antibody of murine or rodent origin. That initial antibody may then be modified, such as using recombinant techniques to produce rodent variants, chimeric antibodies, humanized antibodies and the like. Othermethods exist to produce an initial antibody, and suchmethods are known in the art. However, regardless of the method used to generate an initial antibody or even a variant of that initial antibody, any given antibody of non-human origin can then be modified to increase its humanness.

[0066] Antibodies or antigen-binding fragments of the disclosure canbemadeby using combinatorial libraries to screen for antibodies with the desired activity or activities. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing the desired binding characteristics. Such methods are described generally in Hoogenboom et al. in Methods in Molecular Biology 178:1‑37 (O’Brien et al., ed., Human Press, Totowa, NJ, 2001). For example, one method of generating antibodies of interest is through the use of a phage antibody library as described in Lee et al., J. Mol. Biol. (2004), 340(5):1073‑93.

[0067] In principle, synthetic antibody clones are selected by screening phage libraries containing phage that display various fragments of antibody variable region (Fv) fused to phage coat protein. Such phage libraries are pannedby affinity chromatography against the desired antigen. Clones expressing Fv fragments capable of binding to the desired antigen are adsorbed to the antigen and thus separated from the non-binding clones in the library. The binding clones are then eluted from the antigen, and can be further enriched by additional cycles of antigen adsorption / elution. Any of the antibodies of the disclosure can be obtained by designing a suitable antigen screening procedure to select for the phage clone of interest followed by construction of a full length antibody clone using the Fv sequences from the phage clone of interest and suitable constant region (Fc) sequences described in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91‑3242, Bethesda MD (1991), vols. 1‑3.It can be advantageous to increase the humanness of a non-human antibody to make it more suitable for use in human subject and cells, whether for diagnostic, therapeutic, or research purposes. Antibodies may be modified for use as therapeutics. Examples of such antibodies (including antibody fragments) include chimeric, humanized, and fully human antibodies. Numerousmethods exist in the art for thegenerationof chimeric, humanizedandhumanantibodies. In thecontext of thepresent disclosure, anantibody is considered humanized if at least one of the VH domain or VL domain is humanized. Moreover, a VH or VL domain is humanized if the amino acid sequence of at least a portion of at least one of the FR regions has beenmodified, relative to a parent non-human (e.g., murine) antibody, such that the amino acid sequence of that portion corresponds to that of a human antibody or a human consensus sequence. In certain embodiments, at least one, two, three, or four FR regions of the VHdomain and / or at least one, two, three, or four FR regions of theVL domain have beenmodified (inwhole or in part) so that their sequence is more closely related to a human sequence. For any of the foregoing in certain embodiments, a humanized antibody fragment may be provided in the context of a human or non-human light chain and / or heavy chain constant region (e.g., comprising a CL and one or more of a CH1, hinge, CH2, and / or CH3 domains). In certain embodiments, a humanized antibody or antigen binding fragment of the disclosure is provided in the context of human light and / or heavy chain constant domains, when present. Antibodies and antibody binding fragments combining any of the humanized light chain variable domains and / or heavy chain variable domains described herein are exemplary of antibodies and antigen binding fragments of the disclosure. In some embodiments, the antibody or antigen-binding fragment is humanized. In some embodiments, the antibody or antigen-binding fragment is chimeric. In some embodi- ments, the antibody or antigen-binding fragment is human.

[0068] According tocertainembodimentsof thepresentdisclosure, anti-PAR2antibodiesareprovidedcomprisinganFc domain comprising oneormoremutationswhichenhanceor diminish antibodybinding to theFcRn receptor,e.g.,at acidic pH as compared to neutral pH. For example, the present disclosure includes anti-PAR2 antibodies comprising amutation in the CH2 or a CH3 region of the Fc domain, wherein themutation(s) increases the affinity of the Fc domain to FcRn in an acidic environment (e.g., in an endosomewhere pH ranges from about 5.5 to about 6.0). Suchmutations may result in an increase in serumhalf-life of the antibodywhenadministered to ananimal. Non-limiting examplesof suchFcmodifications 16 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 include, e.g., amodification at position 250 (e.g., E orQ); 250 and 428 (e.g., L or F); 252 (e.g., L / Y / F / Wor T), 254 (e.g., S or T), and 256 (e.g., S / R / Q / E / D or T); or a modification at position 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or 434 (e.g., H / ForY); or amodificationat position250and / or 428; or amodificationat position307or 308 (e.g.,308F,V308F), and434. In one embodiment, the modification comprises a 428L(e.g., M428L) and 434S (e.g., N434S) modification; a 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modification; a 433K (e.g., H433K) and a 434 (e.g., 434Y) modification; a 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modification; a 250Q and 428L modification (e.g., T250Q and M428L); and a 307 and / or 308modification (e.g., 308For308P). In yet another embodiment, themodification comprisesa265A (e.g.,D265A) and / or a 297A (e.g., D297A) modification. All possible combinations of the foregoing Fc domain mutations, and other mutations within the antibody variable domains disclosed herein, are contemplated within the scope of the present disclosure. In some embodiments, antibodies comprise the triple mutation L234F / L235E / P331S ("TM"). TM causes a profound decrease in the binding activity of human IgG1 molecules to human C1q, CD64, CD32A and CD16. See, e.g., Oganesyan et al., ActaCrystallogrDBiol Crystallogr. 64:700‑704 (2008). Antibodieswith increased half-livesmay also be generatedbymodifyingaminoacid residues identifiedas involved in the interactionbetween theFcand theFcRn receptor. For example, the introduction of the triple mutation M252Y / S254T / T256E (’YTE’) into the CH2 domain of human immunoglobulin G (IgG) molecules causes an increase in their binding to the human neonatal Fc receptor (FcRn). See U.S. Patent No. 7.083,784, the contents of which are herein incorporated by reference in its entirety. In some ebodiments, the antibodies comprise the YTE modifications.

[0069] According to certain embodiments of the present disclosure, anti-PAR2 antibodies are provided comprising one ormoremutations in the VHand / or VL domains which enhance or diminish antibody binding to PAR2, e.g., at acidic pH as compared to neutral pH. For example, the present disclosure includes anti-PAR2 antibodies comprising amutation in the CDR2 (SEQ ID NO: 4) or a CDR3 (SEQ ID NO: 5) region of the VH domain and / or the CDR3 (SEQ ID NO: 10) of the VL domain,wherein themutation(s) replaceoneormoreaminoacidswith histidineanddecreases theaffinity of theVHand / or VL domain to PAR2 in an acidic environment (e.g., in an endosome where pH ranges from about 5.5 to about 6.0). Such mutations may result in an increase in serum half-life of the antibody when administered to an animal. Non-limiting examplesof suchVHmodifications include,e.g.,amodification at aminoacid positions4, 5, 7, 8, 10, 11, 12, 14, 15, 16, and 17 of CDR2 (SEQ IDNO: 4) and 1, 2, 4, 5, and 7 of CDR3 (SEQ IDNO: 5). Non-limiting examples of suchVLmodifications include, e.g., a modification at positions 1, 2, 4, 5, 6, 7, 8, 9, 12, and 14 of CDR3 (SEQ ID NO: 10). In yet another embodiment, theVHcomprisesmodifications at positions5, 8, 12, 16, and 17ofCDR2 (SEQ IDNO:4) andpositions 2and 3 of CDR3 (SEQ IDNO: 5). All possible combinations of the foregoing VH and VL domainmutations, and other mutations within the Fc domain disclosed herein, are contemplated within the scope of the present disclosure.

[0070] In some embodiments, the disclosure provides for an antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) anda light chain variable domain (VL), wherein theVHcomprises: i) aVH-CDR1having the amino acid sequence of SEQ IDNO: 3; ii) a VH-CDR2having the amino acid sequence of SEQ IDNO: 4, but wherein a histidine is optionally present at anyoneormoreof theaminoacid positions corresponding to positions1‑17 (e.g.,4, 5, and 7‑17) of SEQ ID NO: 4; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑8 of SEQ ID NO: 5; and wherein theVLcomprises: i) aVL-CDR1having theaminoacidsequenceofSEQIDNO:8; ii) aVL-CDR2having theamino acid sequence of SEQ ID NO: 9; and iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; but wherein a histidine is optionally present at anyoneormoreof theaminoacidpositionscorresponding to positions1‑14ofSEQ IDNO: 10. In some embodiments, the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, ii) a VH- CDR2having theaminoacid sequenceofSEQ IDNO:4, iii) aVH-CDR3having theaminoacid sequenceofSEQ IDNO: 5, and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ IDNO: 8, ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9, iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10. In some embodiments, the antibodyor antigen fragment thereof hasahistidineat theaminoacid position corresponding to anyone ormoreof positions7, 8, 12, 15, 16, or 17ofSEQ IDNO:4. In someembodiments, theantibodyor antigen fragment thereof has a histidine at the amino acid position corresponding to any one or more of positions 2 or 3 of SEQ ID NO: 5. In some embodiments, the antibodyor antigen fragment thereof hasahistidineat theaminoacid position corresponding to anyone ormoreof positions1, 5, 6, or 14ofSEQ IDNO:10. In someembodiments, a histidine is present at theaminoacid positions corresponding to positions 5, 8, 12, 16, and 17 of SEQ ID NO: 4; and wherein a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ IDNO: 5. In some embodiments, the VH-CDR2 comprises an amino acid sequence selected from the group consisting of: SEQ IDNOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794, and 811‑818. In some embodiments, the VH-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 17 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 765, 775, 785, 795, and 819‑820. In some embodiments, the VL-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790 and 800. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 14; wherein the VH-CDR3 comprises an amino acid sequence corresponding toSEQ IDNO: 15, andwherein theVL-CDR3comprisesanaminoacid sequence corresponding toSEQ ID NO: 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ IDNO: 811; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ IDNO: 819, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding toSEQ IDNO: 814; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 820, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 816; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH-CDR2 comprises an amino acid sequence corresponding toSEQ IDNO: 818; the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. In some embodiments, the VH comprises framework regions that are each at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NOs: 803‑806. In some embodiments, the VL comprises framework regions that are each at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ IDNOs: 807‑810. In some embodiments, the VH comprises an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%or 100% identical to any of the sequences selected from the groupconsistingof:SEQIDNOs:2, 12, 22, 32,42, 52, 62, 72, 82,92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, and 792. In some embodiments, the VL comprises an amino acid sequence that is at least 80%,85%,90%,91%,92%,93%,94%,95%,96%,97%,98%,99%or100% identical to anyof the sequencesselected from thegroupconsistingof:SEQ IDNOs:7, 17, 27, 37, 47, 57,67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797. In some embodiments, the VH comprises an amino acid sequence corresponding to SEQ ID NO: 12 and the VL comprises an amino acid sequence corresponding to SEQ ID NO: 17. In some embodiments, the VH comprises an amino acid sequence corresponding to SEQ ID NO: 821 and the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. In some embodiments, the VH comprises an amino acid sequence corresponding to SEQ ID NO: 824 and the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. In some embodiments, the VH comprises an amino acid sequence corresponding toSEQ IDNO:827and theVLcomprisesanaminoacid sequencecorresponding toSEQ IDNO: 7 or 17. In some embodiments, the VH comprises an amino acid sequence corresponding to SEQ ID NO: 831 and the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17.

[0071] The present disclosure also includes anti-PAR2 antibodies comprising a chimeric heavy chain constant (CH) region, wherein the chimeric CH region comprises segments derived from the CH regions of more than one immunoglo- bulin isotype. For example, the antibodies of the disclosuremay comprise a chimeric CH region comprising part or all of a CH2 domain derived froma human IgG1, human IgG2 or human IgG4molecule, combinedwith part or all of aCH3domain derived fromahuman IgG1,human IgG2orhuman IgG4molecule.According tocertainembodiments, theantibodiesof the disclosure comprise a chimericCH region havinga chimeric hinge region. For example, a chimeric hingemaycomprise an "upper hinge" amino acid sequence (amino acid residues from positions 216 to 227 according to EU numbering) derived from a human IgG1, a human IgG2 or a human IgG4 hinge region, combined with a "lower hinge" sequence (amino acid residues frompositions228 to236according toEUnumbering)derived fromahuman IgG1,ahuman IgG2orahuman IgG4 hinge region.

[0072] According to certain embodiments, the chimeric hinge region comprises amino acid residues derived from a human IgG1 or a human IgG4 upper hinge and amino acid residues derived from a human IgG2 lower hinge. An antibody comprising a chimeric CH region as described herein may, in certain embodiments, exhibit modified Fc effector functions without adversely affecting the therapeutic or pharmacokinetic properties of the antibody. (See, e.g., US 2015‑0203591 A1).

[0073] The present disclosure includes anti-PAR2 antibodies or antigen-binding fragments which interact with one or more amino acids of PAR2. The epitope to which the antibodies bind may consist of a single contiguous sequence of 3 or more (e.g.,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20ormore) aminoacidsofPAR2.Alternatively, theepitope may consist of a plurality of non-contiguous amino acids (or amino acid sequences) of PAR2. 18 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55

[0074] Various techniques known to persons of ordinary skill in the art can be used to determine whether an antibody "interactswith oneormoreaminoacids"within apolypeptide or protein.Exemplary techniques include,e.g., routine cross- blocking assay such as that described Antibodies, Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harb., NY), alanine scanningmutational analysis, peptide blots analysis (Reineke, 2004,MethodsMolBiol 248:443‑463), andpeptide cleavageanalysis. Inaddition,methodssuchasepitopeexcision,epitopeextractionandchemicalmodificationofantigens can be employed (Tomer, 2000, Protein Science 9:487‑496). Anothermethod that can be used to identify the amino acids within a polypeptidewith which an antibody interacts is hydrogen / deuteriumexchange detected bymass spectrometry. In general terms, the hydrogen / deuterium exchangemethod involves deuterium-labeling the protein of interest, followed by binding the antibody to the deuterium‑ labeled protein. Next, the protein / antibody complex is transferred to water to allow hydrogen‑ deuterium exchange to occur at all residues except for the residues protected by the antibody (which remain deuterium-labeled). After dissociation of the antibody, the target protein is subjected to protease cleavage and mass spectrometry analysis, thereby revealing the deuterium-labeled residues which correspond to the specific amino acids with which the antibody interacts. See, e.g., Ehring (1999) Analytical Biochemistry 267 (2):252‑259; Engen and Smith (2001) Anal. Chem. 73:256A‑265A.

[0075] The present disclosure further includes anti-PAR2 antibodies or antigen-binding fragments thereof that bind to the same epitope as any of the antibodies or antigen-binding fragments described herein (e.g., an antibody or antigen- binding fragment comprising the amino acid sequences of SEQ IDNOs: 12 and 17). Likewise, the present disclosure also includes anti-PAR2 antibodies and antigen-binding fragments that compete for binding to PAR2with any of the antibodies or antigen-binding fragments described herein (e.g., an antibody or antigen-binding fragment comprising the amino acid sequences of SEQ ID NOs: 12 and 17). The skilled worker can easily determine whether an antibody binds to the same epitope as, or competes for binding with, a reference anti-PAR2 antibody by using routine methods known in the art and exemplified herein. For example, to determine if a test antibody binds to the same epitope as a reference anti-PAR2 antibody of the disclosure, the reference antibody is allowed to bind to aPAR2 protein. Next, the ability of a test antibody to bind to the PAR2 molecule is assessed. If the test antibody is able to bind to PAR2 following saturation binding with the referenceanti-PAR2antibody, it canbeconcluded that the test antibodybinds toadifferent epitope than the referenceanti- PAR2 antibody.On the other hand, if the test antibody is not able to bind to thePAR2molecule following saturation binding with the reference anti-PAR2 antibody, then the test antibody may bind to the same epitope as the epitope bound by the reference anti-PAR2 antibody of the disclosure. Additional routine experimentation (e.g., peptide mutation and binding analyses) can then be carried out to confirm whether the observed lack of binding of the test antibody is in fact due to binding to the same epitope as the reference antibody or if steric blocking (or another phenomenon) is responsible for the lackof observedbinding.Experimentsof this sort canbeperformedusingELISA,RIA,Biacore, flowcytometry or anyother quantitative or qualitative antibody-binding assay available in the art. In accordance with certain embodiments of the present disclosure, two antibodies bind to the same (or overlapping) epitope if, e.g., a 1‑, 5‑, 10‑, 20‑ or 100-fold excess of one antibody inhibits binding of the other by at least 50% but preferably 75%, 90% or even 99% as measured in a competitive binding assay (see, e.g., Junghans et al., Cancer Res. 1990:50:1495‑1502).

[0076] Alternatively, two antibodies are deemed to bind to the same epitope if essentially all amino acidmutations in the antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other. Two antibodies are deemed to have "overlapping epitopes" if only a subset of the amino acidmutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other.

[0077] To determine if an antibody competes for binding (or cross-competes for binding) with a reference anti-PAR2 antibody, the above-described binding methodology is performed in two orientations: In a first orientation, the reference antibody is allowed to bind to a PAR2 protein under saturating conditions followed by assessment of binding of the test antibody to the PAR2 molecule. In a second orientation, the test antibody is allowed to bind to a PAR2 molecule under saturating conditions followed by assessment of binding of the reference antibody to the PAR2 molecule. If, in both orientations, only the first (saturating) antibody is capable of binding to thePAR2molecule, then it is concluded that the test antibody and the referenceantibody compete for binding toPAR2.Aswill be appreciated byapersonof ordinary skill in the art, an antibody that competes for binding with a reference antibody may not necessarily bind to the same epitope as the reference antibody, but may sterically block binding of the reference antibody by binding an overlapping or adjacent epitope.

[0078] Methods for generating monoclonal antibodies, including fully human monoclonal antibodies are known in the art. Any such known methods can be used in the context of the present disclosure to make human antibodies that specifically bind to human PAR2.

[0079] Using VELOCIMMUNE™ technology, for example, or any other known method for generating fully human monoclonal antibodies, highaffinity chimeric antibodies toPAR2are initially isolatedhavingahumanvariable regionanda mouse constant region. As in the experimental section below, the antibodies are characterized and selected for desirable characteristics, includingaffinity, selectivity, epitope, etc. If necessary,mouseconstant regionsare replacedwith adesired human constant region, for example wild-type or modified IgG1 or IgG4, to generate a fully human anti-PAR2 antibody. While the constant region selected may vary according to specific use, high affinity antigen-binding and target specificity 19 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 characteristics reside in the variable region. In certain instances, fully human anti-PAR2 antibodies are isolated directly from antigen-positive B cells.

[0080] The anti-PAR2 antibodies and antibody fragments of the present disclosure encompass proteins having amino acid sequences that may vary from those of the described antibodies but that retain the ability to bind PAR2 (e.g.,SEQ ID NO: 801), or more specifically in some embodiments, a PAR2 tethered ligand (e.g., SEQ ID NO: 802). Such variant antibodies and antibody fragments comprise one or more additions, deletions, or substitutions of amino acids when compared to a parent sequence, but exhibit biological activity that is essentially equivalent to that of the described antibodies. Likewise, the anti-PAR2antibody-encodingDNAsequences of the present disclosure encompass sequences that comprise one or more additions, deletions, or substitutions of nucleotides when compared to the disclosed sequences, but that encode an anti-PAR2 antibody or antibody fragment that is essentially bioequivalent to an anti- PAR2 antibody or antibody fragment of the disclosure. Examples of such variant amino acid and DNA sequences are discussed above.

[0081] Two antibodies or antigen-binding fragments are considered bioequivalent if, for example, they are pharma- ceutical equivalents or pharmaceutical alternatives whose rate and extent of absorption do not show a significant difference when administered at the same molar dose under similar experimental conditions, either single does or multiple dose. Some antibodies or antigen-binding fragments will be considered equivalents or pharmaceutical alter- natives if they are equivalent in the extent of their absorption but not in their rate of absorption and yet may be considered bioequivalent because such differences in the rate of absorption are intentional and are reflected in the labeling, are not essential to the attainment of effective body drug concentrations on, e.g., chronic use, and are considered medically insignificant for the particular drug product studied.

[0082] In some embodiments, two antibodies or antigen-binding fragments are bioequivalent if there are no clinically meaningful differences in their safety, purity, and potency.

[0083] In someembodiments, twoantibodiesorantigen-binding fragmentsarebioequivalent if apatient canbeswitched one or more times between the reference product and the biological product without an expected increase in the risk of adverse effects, including a clinically significant change in immunogenicity, or diminished effectiveness, as compared to continued therapy without such switching.

[0084] In some embodiments, two antibodies or antigen-binding fragments are bioequivalent if they both act by a common mechanism or mechanisms of action for the condition or conditions of use, to the extent that such mechanisms are known.

[0085] Bioequivalencemaybedemonstratedby in vivoand in vitromethods.Bioequivalencemeasures include,e.g., (a) an in vivo test in humans or other mammals, in which the concentration of the antibody or its metabolites is measured in blood, plasma, serum, or other biological fluid as a function of time; (b) an in vitro test that has been correlated with and is reasonably predictive of human in vivo bioavailability data; (c) an in vivo test in humans or other mammals in which the appropriate acute pharmacological effect of the antibody (or its target) is measured as a function of time; and (d) in a well- controlled clinical trial that establishes safety, efficacy, or bioavailability or bioequivalence of an antibody.

[0086] Bioequivalent variants of anti-PAR2 antibodies of the disclosure may be constructed by, for example, making various substitutions of residues or sequences or deleting terminal or internal residues or sequences not needed for biological activity. For example, cysteine residues not essential for biological activity can be deleted or replacedwith other amino acids to prevent formation of unnecessary or incorrect intramolecular disulfide bridges upon renaturation. In other contexts, bioequivalent antibodies or antigen-binding fragments may include anti-PAR2 antibody variants comprising amino acid changes which modify the glycosylation characteristics of the antibodies or antigen-binding fragments, e.g., mutations which eliminate or remove glycosylation.

[0087] The present disclosure, according to certain embodiments, provides anti-PAR2 antibodies or antigen-binding fragments that bind to human PAR2 but not to PAR2 from other species. The present disclosure also includes anti-PAR2 antibodies that bind to human PAR2 and to PAR2 from one or more non-human species. For example, the anti-PAR2 antibodies of the disclosure may bind to human PAR2 and may bind or not bind, as the case may be, to one or more of mouse, rat, guinea pig, hamster, gerbil, pig, cat, dog, rabbit, goat, sheep, cow, horse, camel, cynomolgus monkey, marmoset, rhesus or chimpanzee PAR2. According to certain embodiments, the antibodies or antigen-binding fragments bind to PAR2 in human A549 cells, rat KNRK cells, cynomolgus monkey CYNOM-K1 cells or mouse LL / 2 cells.

[0088] The disclosure encompasses anti-PAR2 monoclonal antibodies conjugated to a therapeutic moiety ("immuno- conjugate"), such as a cytotoxin, a chemotherapeutic, an immunosuppressant or a radioisotope. Examples of suitable cytotoxic agents and chemotherapeutic agents for forming immunoconjugates are known in the art (see for example,WO 05 / 103081).

[0089] In some embodiments, the antibodies of the present disclosure may be monospecific, bi-specific, or multi- specific. Multispecific antibodies may be specific for different epitopes of one target polypeptide or may contain antigen- binding domains specific formore than one target polypeptide.See, e.g.,Tutt et al., 1991, J. Immunol. 147:60‑69; Kufer ei a / ., 2004, Trends Biotechnol. 22:238‑244. The anti-PAR2 antibodies or antigen-binding fragments of the present disclosure can be linked to or co-expressed with another functional molecule, e.g., another peptide or protein. For 20 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 example, an antibody or antigen-binding fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent associationor otherwise) tooneormoreothermolecular entities, suchasanother antibodyor antigen- binding fragment to produce a bi-specific or a multispecific antibody with a second binding specificity. For example, the present disclosure includes bi-specific antibodies wherein one arm of an immunoglobulin is specific for human PAR2 or a fragment thereof, and the other arm of the immunoglobulin is specific for a second therapeutic target or is conjugated to a therapeutic moiety.

[0090] An exemplary bi-specific antibody or antigen-binding fragment format that can be used in the context of the present disclosure involves the use of a first immunoglobulin (Ig) CH3 domain and a second Ig CH3 domain, wherein the first and second Ig CH3 domains differ from one another by at least one amino acid, and wherein at least one amino acid difference reduces binding of the bispecific antibody to Protein A as compared to a bi-specific antibody lacking the amino acid difference. In one embodiment, the first Ig CH3 domain binds Protein A and the second Ig CH3 domain contains a mutation that reduces or abolishes Protein A binding such as an H95Rmodification (by IMGTexon numbering; H435R by EU numbering). The second CH3 may further comprise a Y96F modification (by IMGT; Y436F by EU). Further modifications that may be found within the second CH3 include: D16E, L18M, N44S, K52N, V57M, and V82I (by IMGT; D356E, L358M, N384S, K392N, V397M, and V422I by EU) in the case of lgG1 antibodies; N44S, K52N, and V82I (IMGT; N384S, K392N, and V422I by EU) in the case of IgG2 antibodies; andQ15R, N44S, K52N, V57M, R69K, E79Q, and V82I (by IMGT;Q355R,N384S,K392N,V397M,R409K,E419Q, andV422I byEU) in the caseof IgG4antibodies. Variations on the bi-specific antibody format described above are contemplated within the scope of the present disclosure.

[0091] Other exemplary bispecific formats that can be used in the context of the present disclosure include, without limitation, e.g., scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (DVD)‑lg, Quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEED)body, leucine zipper, Duobody, lgG1 / lgG2, dual acting Fab (DAF)‑lgG, and Mab<2>bispecific formats (see, e.g., Kleinet al. 2012,mAbs4:6, 1 ‑11 , and referencescited therein, for a reviewof the foregoing formats). Bispecificantibodies or antigen-binding fragments can also be constructed using peptide / nucleic acid conjugation, e.g., wherein unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates which then self-assemble intomultimeric complexeswith defined composition, valency and geometry. (See, e.g., Kazane et al., J. Am. C em. Soc. [Epub: Dec. 4, 2012]). C. Nucleic Acids and Expression Systems

[0092] In some embodiments, the disclosure provides for a nucleic acid capable of expressing any of the antibodies of antigen-binding fragments disclosed herein. The nucleic acids may be single-stranded or double-stranded, DNA or RNA molecules. In further embodiments, the antibody or antigen-binding fragment nucleic acid sequences can be isolated, recombinant, and / or fused with a heterologous nucleotide sequence, or in a DNA library. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191,201, 211,221, 231, 241, 251, 261, 271, 281,291, 301, 311,321, 331, 341, 351, 361, 371,381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511,521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, and / or 791. In some embodiments, the nucleic acid comprises a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and / or 796. In particular embodiments, the nucleic acid comprises a nucleotide sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 1, 6, 11 and / or 16

[0093] In certain embodiments, nucleic acids encoding antibodies or antigen-binding fragments also include nucleotide sequences that hybridize under highly stringent conditions to a polynucleotide encoding any of the above-mentioned antibodies or antigen-binding fragments nucleotide sequence, or complement sequences thereof. In someembodiments, thenucleicacidshybridizeunderhighly stringent conditions toapolynucleotideencodinganaminoacidsequence that isat least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of SEQ IDNOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, and 792. In some embodiments, the nucleic acids hybridize under highly stringent conditions to a polynucleotide encoding an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%or 100% identical to any one of SEQ IDNOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 21 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797.One of ordinary skill in the art will understand readily that appropriate stringency conditions which promote DNA hybridization can be varied. For example, one could perform thehybridization at 6.0 x sodiumchloride / sodiumcitrate (SSC) at about 45 °C, followedby awashof 2.0 x SSC at 50 °C. For example, the salt concentration in the wash step can be selected from a low stringency of about 2.0 x SSC at 50 °C to a high stringency of about 0.2 x SSC at 50 °C. In addition, the temperature in the wash step can be increased from low stringency conditions at room temperature, about 22 °C, to high stringency conditions at about 65 °C. Both temperature and salt may be varied, or temperature or salt concentration may be held constant while the other variable is changed. In one embodiment, the disclosure provides nucleic acids which hybridize under low stringency conditions of 6 x SSC at room temperature followed by a wash at 2 x SSC at room temperature.

[0094] Isolated nucleic acids which differ from the nucleic acids encoding the antibody or antigen-binding fragment thereof due to degeneracy in the genetic code are alsowithin the scope of the disclosure. For example, a number of amino acids are designated bymore than one triplet. Codons that specify the same amino acid, or synonyms (for example, CAU and CAC are synonyms for histidine) may result in "silent" mutations which do not affect the amino acid sequence of the protein. However, it is expected that DNA sequence polymorphisms that do lead to changes in the amino acid sequences of the subject proteinswill exist amongmammalian cells.Oneskilled in theartwill appreciate that these variations in oneor more nucleotides (up to about 3‑5%of the nucleotides) of the nucleic acids encoding a particular proteinmay exist among individuals of a given species due to natural allelic variation. Any and all such nucleotide variations and resulting amino acid polymorphisms are within the scope of this disclosure.

[0095] In some embodiments, the disclosure provides for a vector comprising any of the nucleic acids disclosed herein. In some embodiments, the disclosure provides for a host cell comprising any of the vectors disclosed herein.

[0096] Regardless of when an antibody of the disclosure is a full length antibody or an antigen binding fragment, antibodies and antigen binding fragments of the disclosure can be recombinantly expressed in cell lines. In these embodiments, sequences encoding particular antibodies or antigen binding fragments can beused for transformation of a suitable host cell, such as amammalian host cell or yeast host cell. According to these embodiments, transformation can beachievedusing any knownmethod for introducing polynucleotides into ahost cell, including, for example packaging the polynucleotide in a virus (or into a viral vector) and transducing a host cell with the virus (or vector) or by transfection procedures known in the art. Generally, the transformation procedure usedmay depend upon the host to be transformed. Methods for introducing heterologous polynucleotides intomammalian cells are well known in the art and include, but are not limited to, dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, proto- plast fusion, electroporation, encapsulation of thepolynucleotide(s) in liposomes, anddirectmicroinjectionof theDNA into nuclei.

[0097] According to certain embodiments of the disclosure, a nucleic acidmolecule encoding the amino acid sequence of a heavy chain constant region (all or a portion), a heavy chain variable region of the disclosure, a light chain constant region, or a light chain variable region of the disclosure is inserted into an appropriate expression vector using standard ligation techniques. In a preferred embodiment, the heavy or light chain constant region is appended to the C-terminus of the appropriate variable region and is ligated into an expression vector. The vector is typically selected to be functional in the particular host cell employed (i.e., the vector is compatible with the host cell machinery such that amplification of the gene and / or expression of the gene can occur). For a review of expression vectors, see, Goeddel (ed.), 1990, Meth. Enzymol. Vol. 185, Academic Press. N.Y. In the context of antibody expression, both the heavy and light chain may be expressed from the same vector (e.g., from the same or different promoters present on the same vector) or the heavy and light chains may be expressed from different vectors. In certain embodiments, the heavy and light chains are expressed from different vectors which are transfected into the same host cell and co-expressed. Regardless of when the heavy and light chainsareexpressed in thesamehost cell from thesameoradifferent vector, thechainscan thenassociate to forman antibody (or antibody fragment, depending on the portions of the heavy and light chain being expressed).

[0098] Typically, expressionvectors used in anyof thehost cellswill contain sequences for plasmidmaintenanceand for cloning and expression of exogenous nucleotide sequences. Such sequences, collectively referred to as "flanking sequences" in certain embodiments will typically include one or more of the following nucleotide sequences: a promoter, one or more enhancer sequences, an origin of replication, a transcriptional termination sequence, a complete intron sequence containing a donor and acceptor splice site, a sequence encoding a leader sequence for polypeptide secretion, a ribosome binding site, a polyadenylation sequence, a polylinker region for inserting the nucleic acid encoding the polypeptide to be expressed, and a selectable marker element. These portions of vectors are well known, and there are numerous generally available vectors that can be selected and used for the expression of proteins. One can readily select vectors based on the desired host cell and application.

[0099] Anorigin of replication is typically apart of thoseprokaryotic expressionvectorspurchasedcommercially, and the origin aids in the amplification of the vector in a host cell. If the vector of choice does not contain an origin of replication site, one may be chemically synthesized based on a known sequence, and ligated into the vector. For example, the origin of replication from the plasmid pBR322 (New England Biolabs, Beverly, Mass.) is suitable for most gram-negative bacteria 22 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 and various viral origins (e.g., SV40, polyoma, adenovirus, vesicular stomatitus virus (VSV), or papillomaviruses such as HPVorBPV)areuseful for cloningvectors inmammaliancells.Generally, theoriginof replicationcomponent is notneeded formammalian expression vectors (for example, theSV40origin is often used only because it also contains the virus early promoter).

[0100] The expression and cloning vectors of the disclosure will typically contain a promoter that is recognized by the host organism and operably linked to the molecule encoding heavy and / or light chain. Promoters are untranscribed sequences located upstream (i.e., 5’) to the start codon of a structural gene (generally within about 100 to 1000 bp) that control the transcription of the structural gene. Promoters are conventionally grouped into one of two classes: inducible promoters and constitutive promoters. Inducible promoters initiate increased levels of transcription fromDNA under their control in response to some change in culture conditions, such as the presence or absence of a nutrient or a change in temperature. Constitutive promoters, on the other hand, initiate continual gene product production; that is, there is little or no control over gene expression. A large number of promoters, recognized by a variety of potential host cells, are well known.Asuitablepromoter is operably linked to theDNAencoding theheavychainor light chain comprisinganantibodyor antigenbinding fragment of thedisclosure. In certain embodiments, the samepromoter is used for both theheavy and light chain. In other embodiments, different promoters (present on the same or different vectors) are used for each.

[0101] Suitable promoters for use with yeast hosts are also well known in the art. Yeast enhancers are advantageously used with yeast promoters. Suitable promoters for use with mammalian host cells are well known and include, but are not limited to, those obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, retroviruses, hepatitis-B virus and most preferablySimianVirus40 (SV40).Other suitablemammalianpromoters includeheterologousmammalianpromoters, for example, heat-shock promoters and the actin promoter.

[0102] Additional promoters which may be of interest include, but are not limited to: the SV40 early promoter region (Bernoist and Chambon, 1981, Nature 290:304‑10); the CMV promoter; the promoter contained in the 3’ long terminal repeat of Rous sarcoma virus (Yamamoto et al., 1980, Cell 22:787‑97); the herpes thymidine kinase promoter (Wagner et al., 1981, Proc. Natl. Acad. Sci. USA 78:1444‑45); the regulatory sequences of the metallothionine gene (Brinster et al., 1982, Nature 296:39‑42); prokaryotic expression vectors such as the beta-lactamase promoter (Villa-Kamaroff et al., 1978, Proc. Natl. Acad. Sci. USA 75:3727‑31); or the tac promoter (DeBoer et al., 1983, Proc. Natl. Acad. Sci. USA 80:21‑25). Also of interest are the following animal transcriptional control regions,which exhibit tissue specificity and have been utilized in transgenic animals: the elastase I gene control region that is active in pancreatic acinar cells (Swift et al., 1984, Cell 38:639‑46; Ornitz et al., 1986, Cold Spring Harbor Symp. Quant. Biol. 50:399‑409 (1986); MacDonald, 1987, Hepatology 7:425‑515); the insulin gene control region that is active in pancreatic beta cells (Hanahan, 1985, Nature 315:115‑22); the immunoglobulin gene control region that is active in lymphoid cells (Grosschedl et al., 1984, Cell 38:647‑58; Adames et al., 1985, Nature 318:533‑38; Alexander et al., 1987, Mol. Cell. Biol. 7:1436‑44); the mouse mammary tumor virus control region that is active in testicular, breast, lymphoid and mast cells (Leder et al., 1986, Cell 45:485‑95); the albumin gene control region that is active in liver (Pinkert et al., 1987, Genes and Devel. 1:268‑76); the alpha-feto-proteingenecontrol region that is active in liver (Krumlauf et al., 1985,Mol.Cell. Biol. 5:1639‑48;Hammeret al., 1987, Science 235:53‑58); the alpha 1-antitrypsin gene control region that is active in liver (Kelsey et al., 1987,Genes and Devel. 1:161‑71); the beta-globin gene control region that is active in myeloid cells (Mogram et al., 1985, Nature 315:338‑40; Kollias et al., 1986, Cell 46:89‑94); the myelin basic protein gene control region that is active in oligoden- drocyte cells in the brain (Readheadet al., 1987,Cell 48:703‑12); themyosin light chain‑2genecontrol region that is active in skeletal muscle (Sani, 1985, Nature 314:283‑86); and the gonadotropic releasing hormone gene control region that is active in the hypothalamus (Mason et al., 1986, Science 234:1372‑78).

[0103] The vector may also include an enhancer sequence to increase transcription of DNA encoding light chain or heavy chain.

[0104] Expression vectors of the disclosuremay be constructed froma starting vector such as a commercially available vector. Such vectors may or may not contain all of the desired flanking sequences. Where one or more of the flanking sequences described herein are not already present in the vector, they may be individually obtained and ligated into the vector. Methods used for obtaining each of the flanking sequences are well known to one skilled in the art.

[0105] After the vector has been constructed and a nucleic acid molecule encoding light chain or heavy chain or light chain and heavy chain comprising an antibody or antigen binding fragment of the disclosure has been inserted into the proper site of the vector, the completed vectormay be inserted into a suitable host cell for amplification and / or polypeptide expression. The transformation of an expression vector into a selected host cell may be accomplished by well known methods including transfection, infection, calciumphosphate co-precipitation, electroporation,microinjection, lipofection, DEAE-dextranmediated transfection, or other known techniques. Themethod selectedwill in part be a function of the type of host cell to be used. These methods and other suitable methods are well known to the skilled worker.

[0106] The host cell, when cultured under appropriate conditions, synthesizes the antibody or antigen binding fragment of the disclosure that can subsequently be collected from the culturemedium (if the host cell secretes it into themedium) or directly from the host cell producing it (if it is not secreted). The selection of an appropriate host cell will depend upon 23 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 various factors, such as desired expression levels, polypeptide modifications that are desirable or necessary for activity (such as glycosylation or phosphorylation) and ease of folding into a biologically active molecule.

[0107] Mammalian cell lines available as host cells for expression are well known in the art and include, but are not limited to, many immortalized cell lines available from the American Type Culture Collection (A.T.C.C.), including but not limited to Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), and a number of other cell lines. In another embodiment, one may select a cell line from the B cell lineage that does not make its own antibody but has a capacity to make and secrete a heterologous antibody (e.g., mouse myeloma cell lines NS0 and SP2 / 0). In other embodiments, a cell other than a mammalian cell is used, such as a yeast cell line (e.g., Pichia).

[0108] In certain embodiments, the cell line stably expresses an antibody or antigen binding fragment of the disclosure. In other embodiments, the cells transiently express an antibody or antigen binding fragment of the disclosure. D. Therapeutic Formulation and Administration

[0109] The disclosure provides pharmaceutical compositions comprising the anti-PAR2 antibodies or antigen-binding fragments thereof of the present disclosure. The pharmaceutical compositions of the disclosure are formulated with suitable carriers, excipients, and other agents that provide improved transfer, delivery, tolerance, and the like. Amultitude of appropriate formulations can be found in the formulary known to all pharmaceutical chemists: Remington’s Pharma- ceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LIPOFECTIN™, Life Technologies, Carlsbad, CA), anhydrous absorption pastes, oil-in-water andwater-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al. "Compendium of excipients for parenteral formulations" PDA (1998) J Pharm Sci Technol 52:238‑31 1.

[0110] The dose of antibody administered to a patient may vary depending upon the age and the size of the patient, targetdisease, conditions, routeofadministration,and the like.Thepreferreddose is typically calculatedaccording tobody weight or body surface area. Depending on the severity of the condition, the frequency and the duration of the treatment can be adjusted. Effective dosages and schedules for administering anti-PAR2 antibodies or antigen-binding fragments may be determined empirically; for example, patient progress can be monitored by periodic assessment, and the dose adjusted accordingly. Moreover, interspecies scaling of dosages can be performed using well-known methods in the art (e.g., Mordenti et al., 1991 , Pharmaceut. Res. 8:1351).

[0111] Various delivery systems are known and can be used to administer the pharmaceutical composition of the disclosure, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the mutant viruses, receptor mediated endocytosis (see, e.g., Wu et al., 1987, J. Biol. Chem. 262:4429‑4432). Methods of introduction include, but are not limited to, intradermal, intrathecal, intramuscular, intraperitoneal, intravenous, subcu- taneous, intranasal, epidural, andoral routes. Thecompositionmaybeadministeredbyany convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local.

[0112] In some embodiments, the antibodies and antigen-binding fragments thereof have utility in treating conditions anddisordersassociatedwith thecentral nervoussystem,and,particularlyassociatedwith thebrain.Whilevarious factors must be considered when administering amacromolecule such as an antibody or antigen-binding fragment to a subject’s brain, i.e., ability of the antibody or antigen-binding fragment to cross the blood-brain-barrier (BBB), the skilled worker is awareofmethodsof administering suchmacromolecules to thebrain. For example, in someembodiments, theantibodyor antigen-binding fragment is covalentlymodifiedwith one ormore cationic polyamines, such as hexamethylenediamine or tetramethylenediamine in order to increase the likelihood that the antibody or antigen-binding fragment is internalized across the BBB. In some embodiments, the antibody or antigen-binding fragment is a bispecific antibody or antigen- binding fragment, wherein the antibody or fragment targets PAR2 and also targets a receptor that facilitates transport across the BBB (e.g., transferrin receptor, insulin receptor and TMEM30A). In some embodiments, the antibody or antigen-binding fragment is conjugated to an agent that targets a receptor that facilitates transport across the BBB (e.g., transferrin receptor, insulin receptor and TMEM30A). In some embodiments, the BBB is temporarily disrupted prior to or during administration of the antibody or fragment. In some embodiments the BBB is temporarily disrupted by means of ultrasound, radiation, biochemical treatment (e.g., with a KCa receptor agonist such as NS‑1619), or intra-arterial infusion of concentrated hyperosmotic solutions.

[0113] Apharmaceutical composition of the present disclosure canbe delivered subcutaneously or intravenouslywith a standard needle and syringe. In addition, with respect to subcutaneous delivery, a pen delivery device readily has applications in delivering a pharmaceutical composition of the present disclosure. Such a pen delivery device can be reusable or disposable. A reusable pen delivery device generally utilizes a replaceable cartridge that contains a pharmaceutical composition. Once all of the pharmaceutical composition within the cartridge has been administered 24 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 and thecartridge isempty, theempty cartridge can readily bediscardedand replacedwith anewcartridge that contains the pharmaceutical composition. The pen delivery device can then be reused. In a disposable pen delivery device, there is no replaceable cartridge. Rather, the disposable pen delivery device comes prefilled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0114] Numerous reusable pen and autoinjector delivery devices have applications in the subcutaneous delivery of a pharmaceutical composition of the present disclosure. Examples include, but are not limited to AUTOPEN™ (Owen Mumford, Inc., Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX 75 / 25™ pen, HUMALOG™ pen, HUMALIN 70 / 30™ pen (Eli Lilly and Co., Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™,OPTIPENPRO™, OPTIPENSTARLET™, andOPTICLIK™ (sanofi- aventis, Frankfurt, Germany), to name only a few. Examples of disposable pen delivery devices having applications in subcutaneous delivery of a pharmaceutical composition of the present disclosure include, but are not limited to the SOLOSTAR™ pen (sanofi-aventis), the FLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™ Autoinjector (Amgen,ThousandOaks,CA), thePENLET™(Haselmeier,Stuttgart,Germany), theEPIPEN(Dey, L.P.), and the HUMIRA™ Pen (Abbott Labs, Abbott Park IL), to name only a few.

[0115] In certain situations, the pharmaceutical composition can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, polymericmaterials can be used; see, Medical Applications of Controlled Release, Langer andWise (eds.), 1974,CRCPres., BocaRaton, Florida. In yet another embodiment, a controlled release systemcan beplaced in proximity of the composition’s target, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, 1984, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115‑138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527‑1533.

[0116] The injectable preparations may include dosage forms for intravenous, subcutaneous, intrathecal, intracuta- neous and intramuscular injections, drip infusions, etc. These injectable preparations may be prepared by methods publicly known. For example, the injectable preparationsmay be prepared, e.g., by dissolving, suspending or emulsifying anyof theantibodiesorantigen-binding fragmentsdisclosedhereinor its salt describedabove inasterileaqueousmedium or an oily medium conventionally used for injections. As the aqueous medium for injections, there are, for example, physiological saline, an isotonic solution containing glucose and other auxiliary agents, etc., which may be used in combinationwith anappropriate solubilizing agent suchasanalcohol (e.g.,ethanol), a polyalcohol (e.g.,propyleneglycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO‑50 (polyoxyethylene (50 mol) adduct of hydro- genated castor oil)], etc. As the oilymedium, there are employed, e.g., sesame oil, soybean oil, etc., whichmay be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared is preferably filled in an appropriate ampoule.

[0117] Advantageously, the pharmaceutical compositions for oral or parenteral use described above are prepared into dosage forms in a unit dose suited to fit a dose of the active ingredients. Such dosage forms in a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the aforesaid antibody contained is generally about 5 to about 500 mg per dosage form in a unit dose; especially in the form of injection, it is preferred that theaforesaidantibodyor antigen-binding fragment is contained inabout5 toabout100mgand inabout 10 to about 250 mg for the other dosage forms. E. Therapeutic Uses of the Antibodies

[0118] For any of themethodsdescribed herein, the disclosure contemplates theuseof anyof the antibodies or antigen- binding fragments of the disclosure.

[0119] In someembodiments, the disclosure provides for amethod of treating a disorder in a subject in which undesired and / or aberrant PAR2 activity is involved, comprising administering any of the antibodies or antigen-binding fragments described herein. As used herein, "disorder", "condition" and "disease" are used interchangeably to refer to any of the disorders, conditions or diseases disclosed herein. In some embodiments, the disease / disorder / condition in which undesired and / or aberrant PAR2 activity is involved is a disease / disorder / condition associatedwith aberrant or undesired inflammation. Examples of diseases / disorders / conditions inwhich aberrant or undesiredPAR2activity is involved include acute or chronic pain, acute or chronic itch, acute or chronic inflammation (e.g., acute or chronic inflammation of the joints, lungs, brain, gastrointestinal tract, periodontium, skin, and vascular systems), autoimmune disorders, periodontitis, osteoarthritis, rheumatoid arthritis, inflammatory bowel disease, arthritis, psoriasis, obesity, diabetes, cardiovascular disease, pancreatitis, cancer (e.g., breast, lung, colon, stomach or prostate cancer), asthma, fibrosis, gastric ulcers, fibrosis or fibrotic disorders, Alzheimer’s Disease, Parkinson’s Disease, contract dermatitis, Crohn’s Disease, ulcerative colitis, adult respiratory distress syndrome (ARDS), glomerulonephritis, and meningitis. In some embodiments, the disclosure provides for methods of treating a subject with metabolic syndrome, or one ormore conditions associated with 25 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 metabolic syndrome, such as visceral fat deposition, hypertension, impaired glucose and insulin homeostasis, insulin resistance, endothelial damage, cardiovascular hypertrophy, inflammation, vascular inflammation, atherosclerosis, ventricular contractile dysfunction, fibrosis and fatty liver disease. In particular embodiments, the disclosure provides for methods of treating pain, e.g., pain associated with any of the diseases / disorders / conditions disclosed herein (e.g., osteoarthritic pain). In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0120] In someembodiments, thedisclosureprovides for amethodof interferingwith the interactionbetweenaprotease (e.g., trypsin) and PAR2, comprising the step of administering any of the antibodies or antigen-binding fragments described herein to a cell. In some embodiments, the disclosure provides for a method of inhibiting exposure of the tethered ligand of PAR2 on a cell, comprising the step of administering any of the antibodies or antigen-binding fragments describedherein to a cell. In someembodiments, thedisclosure provides for amethodof inhibiting the interaction between the tethered ligandofPAR2and thesecond transmembrane loopof thePAR2protein, comprising thestepof administering any of the antibodies or antigen-binding fragments described herein to a cell. In some embodiments, the disclosure provides for a method of inhibiting activation of a PAR2 receptor on a cell, comprising the step of administering any of the antibodies or antigen-binding fragments described herein to a cell. In some embodiments, the cell is a neuron (e.g., a sensory neuron). In some embodiments, the cell is in vitro. In other embodiments, the cell is in a subject. In some embodiments, the subject is amammal. In someembodiments, the subject is a human. In someembodiments, the subject suffers from any of disorders disclosed herein.

[0121] Foranyof themethodsdescribedherein, thedisclosurecontemplates thecombinationof any stepor stepsof one methodwith any step or steps fromanothermethod. Thesemethods involve administering to an individual in need thereof an effective amount of a compound of the disclosure appropriate for the particular disease or condition. In specific embodiments, thesemethods involvedeliveringanyof theantibodiesor antigen-binding fragmentsdisclosedherein to the cells of a subject in need thereof.

[0122] The terms "treatment", "treating", "alleviation" and the like are used herein to generallymean obtaining a desired pharmacologic and / or physiologic effect, and may also be used to refer to improving, alleviating, and / or decreasing the severity of one or more symptoms of a condition being treated. The effect may be prophylactic in terms of completely or partially delaying the onset or recurrence of a disease, condition, or symptoms thereof, and / ormaybe therapeutic in terms of a partial or complete cure for a disease or condition and / or adverse effect attributable to the disease or condition. "Treatment" as used herein covers any treatment of a disease or condition of a mammal, particularly a human, and includes any one ormore of : (a) preventing the disease or condition fromoccurring in a subjectwhichmaybepredisposed to thediseaseor conditionbuthasnot yetbeendiagnosedashaving it; (b) inhibiting thediseaseor condition (e.g., arresting its development); or (c) relieving the disease or condition (e.g., causing regression of the disease or condition, providing improvement in one or more symptoms). For example, "treatment" of pain (e.g., osteoarthritic pain) involves a reduction, arrest, alleviation, or elimination of pain symptoms in the treated subject. The population of subjects treated by themethod of the disease includes subjects suffering from the undesirable condition or disease, as well as subjects at risk for development of the condition or disease.

[0123] For anyof themethodsdescribedherein, thedisclosure contemplates theuseof anyof theantibodies or antigen- binding fragments described throughout the application. In addition, for any of the methods described herein, the disclosure contemplates the combination of any step or steps of onemethod with any step or steps from another method.

[0124] In certain embodiments, the present invention providesmethods of treating conditions associatedwith any of the diseases / conditions / disorders disclosed herein, e.g., acute or chronic pain (e.g., osteoarthritic pain). These methods involve administering to the individual a therapeutically effective amount of any of the antibodies or antigen-binding fragments as described above. These methods are particularly aimed at therapeutic and prophylactic treatments of animals, andmoreparticularly, humans.Thedisclosure contemplatesall combinationsof anyof the foregoingaspectsand embodiments, as well as combinations with any of the embodiments set forth in the detailed description and examples.

[0125] By the term "therapeutically effective dose" is meant a dose that produces the desired effect for which it is administered. Theexact dosewill dependon thepurposeof the treatment, andwill beascertainablebyoneskilled in theart using known techniques (see, e.g., Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

[0126] In certain embodiments, any of the antibodies or antigen-binding fragments of the present invention can be administered alone or in combination with one or more additional compounds or therapies for treating any of the diseases / conditions / disorders disclosed herein, e.g., acute or chronic pain (e.g., osteoarthritic pain). For example, any of the antibodies or antigen-binding fragments disclosed herein can be co-administered in conjunction with one or more therapeutic compounds. When co-administration is indicated, the combination therapy may encompass simultaneous or alternating administration. In addition, the combinationmay encompass acute or chronic administration. Optionally, the antibody / antigen-binding fragment and additional compounds act in an additive or synergistic manner for treating any of the diseases / conditions / disorders disclosed herein, e.g., acute or chronic pain (e.g., osteoarthritic pain). Additional compounds to be used in combination therapies include, but are not limited to, small molecules, polypeptides, antibodies, antisenseoligonucleotides, and siRNAmolecules. In someembodiments, the additional compound is anyone or more of: an anti-inflammatory drug, analgesic, a nonsteroidal anti-inflammatory drug (NSAID), corticosteroid, hya- 26 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 luronic acid, acetaminophen, codeine, lorcet, lortab, vicodin, hydrocodone, morphine, oxycontin, Roxicodone, Percocet, aspirin, celecoxib, pregabalin, joint fusion, joint replacement, abatacept, adalimumab, anakinra, certolizumab, etaner- cept, golimumab, infliximab, rituximab, tocilizumab and tofacitinib. Depending on the nature of the combinatory therapy, administration of the antibodies or antigen binding disclosures of the disclosuremay be continuedwhile the other therapy is being administered and / or thereafter. Administration of the antibodies or antigen-binding fragments may be made in a single dose, or in multiple doses. In some instances, administration of the antibodies or antigen binding fragments is commenced at least several days prior to the other therapy, while in other instances, administration is begun either immediately before or at the time of the administration of the other therapy. In some embodiments, any of the additional compounds disclosed herein is conjugated to any of the antibodies or antigen-binding fragments disclosed herein.

[0127] In another example of combination therapy, any of the antibodies or antigen-binding fragments of the disclosure can be used as part of a therapeutic regimen combined with one or more additional treatment modalities. By way of example, such other treatment modalities include, but are not limited to, dietary therapy, occupational therapy, physical therapy, psychiatric therapy, massage, acupuncture, acupressure, mobility aids, assistance animals, and the like.

[0128] Note that although theantibodiesor antigen-binding fragmentsdescribedherein canbeused in combinationwith other therapies, in certain embodiments, an antibody or antigen-binding fragment is provided as the sole form of therapy. Regardlessofwhetheradministratedaloneor in combinationwithothermedicationsor therapeutic regiments, thedosage, frequency, route of administration, and timing of administration of the antibodies or antigen-binding fragments is determined by a physician based on the condition and needs of the patient.

[0129] According to certain embodiments of the present disclosure,multiple doses of an anti-PAR2antibody or antigen- binding fragment thereof (or a pharmaceutical composition comprising a combination of an anti-PAR2antibody andany of the additional therapies mentioned herein) may be administered to a subject over a defined time course. The methods according to this aspect of the disclosure comprise sequentially administering to a subject multiple doses of an anti-PAR2 antibodyorantigen-binding fragmentof thedisclosure.Asusedherein, "sequentially administering"means that eachdose of anti-PAR2 antibody or antigen-binding fragment is administered to the subject at a different point in time, e.g., on differentdaysseparatedbyapredetermined interval (e.g., hours,days,weeksormonths).Thepresentdisclosure includes methodswhich comprise sequentially administering to thepatient a single initial doseof ananti-PAR2antibody or antigen- binding fragment, followed by one or more secondary doses of the anti-PAR2 antibody or antigen-binding fragment, and optionally followed by one or more tertiary doses of the anti-PAR2 antibody or antigen-binding fragment.

[0130] The terms "initial dose," "secondary doses," and "tertiary doses," refer to the temporal sequence of adminis- tration of the anti-PAR2 antibody or antigen-binding fragment of the disclosure. Thus, the "initial dose" is the dosewhich is administeredat thebeginningof the treatment regimen (also referred toas the "baselinedose"); the "secondarydoses" are the doseswhich are administered after the initial dose; and the "tertiary doses" are the doseswhich are administered after the secondary doses. The initial, secondary, and tertiary dosesmay all contain the same amount of anti-PAR2 antibody or antigen-binding fragment, but generally may differ from one another in terms of frequency of administration. In certain embodiments, however, the amount of anti-PAR2 antibody or antigen-binding fragment contained in the initial, secondary and / or tertiary doses varies from one another (e.g., adjusted up or down as appropriate) during the course of treatment. In certain embodiments, two ormore (e.g., 2, 3, 4, or 5) doses are administered at the beginning of the treatment regimen as "loadingdoses" followedbysubsequent doses that areadministeredona less frequent basis (e.g., "maintenancedoses"). F. Diagnostic / Other Uses of the Antibodies or Antigen Binding Fragments

[0131] The anti-PAR2 antibodies of the present disclosuremay also be used to detect and / or measure PAR2, or PAR2- expressing cells in a sample, e.g., for diagnostic purposes. For example, an anti-PAR2 antibody, or antigen-binding fragment thereof, may be used to diagnose a condition or disease characterized by aberrant expression (e.g., over- expression, under-expression, lack of expression, etc.) of PAR2. Exemplary diagnostic assays for PAR2 may comprise, e.g., contacting a sample obtained from a patient, with an anti-PAR2 antibody of the disclosure, wherein the anti-PAR2 antibody is labeled with a detectable label or reporter molecule.

[0132] Alternatively, an unlabeled anti-PAR2 antibody can be used in diagnostic applications in combination with a secondary antibody which is itself detectably labeled. The detectable label or reporter molecule can be a radioisotope, such as 3H, 14C, 32P, 35S, or 125I; a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine; or an enzyme such as alkaline phosphatase, beta-galactosidase, horseradish peroxidase, or luciferase. Specific exemplary assays that can be used to detect or measure PAR2 in a sample include enzyme-linked immuno- sorbent assay (ELISA), radioimmunoassay (RIA), and fluorescence-activated cell sorting (FACS).

[0133] The compositions of the disclosure have numerous uses. For example, the antibodies and antigen binding fragments of the disclosure are useful for studying preferential cell and tissue distribution in cells and in tissues in vitro and / or in vivo. Similarly, the antibodies and antigen binding fragments, either alone or conjugated to a heterologous agent are useful as imaging agents, such as for ex vivo or in vivo diagnostic applications. For example, the antibodies or antigen binding fragments conjugated to a radioactive moiety are useful for ex vivo or in vivo imaging studies. Similarly, any of the 27 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 antibodies or antigen binding fragments of the disclosure are similarly useful.

[0134] When used in vitro, the antibodies and antigen binding fragments of the disclosure are suitable for identifying binding partners for the antibody or antigen binding fragment being delivered (e.g., identifying proteins or peptides that bind the antibody or antigen binding fragment), and for evaluating localization and trafficking. Similarly, when used in vivo, the antibodies or antigen-binding fragments are useful for identifying binding partners for the antibody or antigen-binding fragment being delivered (e.g., identifying proteins or peptides that bind the antibody or antigen-binding fragment), for evaluating localization and trafficking, for evaluating biodistribution and half-life, and for evaluating immunogenicity. G. Animal / Cell Models

[0135] Numerousanimalmodelsare known to theskilledworker thatwouldbeuseful for examininganyof theantibodies or fragments thereof. See, e.g., Kuyinu et al., 2016, J Orthop Surg Res, 11(19): 10.1186 / s13018‑016‑0346‑5. In some embodiments, the animal model is a pain-based model generated by treatment of the animal with a chemical, such as sodium monoiodoacetate (MIA) or carrageenan. In some embodiments, the chemical is injected at the site of where the pain is to be induced in the animal. In some embodiments, the animal model is an animal in which an injury is introduced postoperatively (e.g., incisional), such as anterior cruciate ligament transection, meniscectomy, or medial meniscal transection. In some embodiments, the animal model is one associated with an inflammatory condition, such as lower esophageal irritation, colon inflammation, stomach ulceration, urinary bladder inflammation, pancreatic inflammation and uterine inflammation. See, e.g., the animal models referred to in National Research Council Committee on Recognition and Alleviation of Pain in Laboratory Animals, "Models of Pain," 2009. H. Kits

[0136] In certain embodiments, the invention also provides a pharmaceutical package or kit comprising one or more containers filled with at least one antibody or antigen-binding fragment of the disclosure. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects (a) approval by the agency of manufacture, use or sale for human administration, (b) directions for use, or both. EXAMPLES

[0137] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how tomake and use themethods and compositions of the disclosure, and are not intended to limit the scopeofwhat the inventors regardas their disclosure.Effortshavebeenmade toensureaccuracywith respect tonumbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric. Example 1: Generation of antibodies with pH sensitive binding for PAR2 Generation of recombinant human, rat and cynomolgus PAR2 and PAR1 proteins.

[0138] The human, rat and cynomolgus (Macaca fascicularis) PAR2 (Proteinase-Activated Receptor 2) constructs comprising extracellular residues 1‑75were designedwithN-terminal AviTag™ (Avidity LLC) andC-terminal Flag andpoly Histidine tagsandcloned into thevectorpDEST12.2OriPFH(LifeTechnologies). ThehumanPAR1(Proteinase-Activated Receptor 1) construct comprising extracellular residues 1‑102 was designed with C-terminal Flag and poly Histidine tags and cloned into the vector pDEST12.2 OriP FH (Life Technologies). The constructs were expressed in HEK293 cells and purified from the media using standard affinity and size exclusion chromatography purification. To generate biotinylated proteins the AviTag™ was biotinylated enzymatically according to the manufacturer’s instructions. Construction of combinatorial histidine scanning libraries

[0139] Split pool oligonucleotides were designed to introduce histidine or the wild type amino acid at each position of either the VHCDR2, VHCDR3 or VLCDR3 of Par0067. Three Par0067 scFv phage display libraries were subsequently constructed in which there were between 0% and 100% histidine residues in either the VHCDR2, VHCDR3 or VLCDR3. 28 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 Selection of pH sensitive Par0067 variant scFv’s

[0140] The combinatorial histidine scanning libraries were subjected to affinity based phage display selections with the aim of isolating Par0067 variants which bind to PAR2 at pH 7.4 but with reduced binding at pH 6.0. To achieve this, four rounds of selectionwere performedwith each library using decreasing concentrations of biotinylated recombinant human PAR2 (Hawkins, RE et al., 1992 Aug 5;226(3):889‑96). At each round, phage were pre-incubated for 1 hour with streptavidin coated paramagnetic beads (dynabeads®) to remove any streptavidin binders. The streptavidin beads were subsequently removedwithaDYNAL®magnet anddiscardedand the remainingphageadded tobiotinylated recombinant human PAR2 at pH 7.4. The selection proceeded for 2 hours before adding streptavidin coated paramagnetic beads to capture the biotinylated recombinant human PAR2 bound phage. The beads were washed 5-times with PBS Tween (PBST) prior to elution of specific scFv in low pH buffer (pH 5.5 - pH 6.0). The selected scFv-phage particles were then rescued as described previously (Osbourn JK.et al. Immunotechnology, 2(3):181‑96, 1996), and the selection process was repeated in the presence of decreasing concentrations of biotinylated PAR2 (1 nM - 0.05 nM over 4 rounds). Reformatting of scFv to IgG1-TM

[0141] Antibodies were converted from scFv to whole immunoglobulin G1 triple mutant (IgG1-TM, IgG1 Fc sequence incorporatingmutations L234F, L235E and P331S) antibody format essentially as described by Persic et al. (1997, Gene, 187, 9‑18) with the following modifications. An OriP fragment was included in the expression vectors to facilitate use with CHO-transient cells and toallowepisomal replication.Thevariableheavy (VH)domainwascloned intoavector containing the human heavy chain constant domains and regulatory elements to expresswhole IgG1-TMheavy chain inmammalian cells. Similarly, the variable light (VL) domainwas cloned into a vector for the expression of the human light chain (lambda) constant domains and regulatory elements to expresswhole IgG light chain inmammalian cells. To obtain IgGs, the heavy and light chain IgG expressing vectors were transfected into CHO-transient mammalian cells (Daramola et al. Biotechnol Prog30(1):132‑41 (2014)). IgGswereexpressedandsecreted into themedium.Harvestswerefilteredprior topurification, then IgGwas purified usingProtein A chromatography. Culture supernatants were loaded on a columnof appropriate size of Ceramic Protein A (BioSepra) and washed with 50 mM Tris-HCl pH 8.0, 250mMNaCl. Bound IgGwas eluted from the column using 0.1 M Sodium Citrate (pH 3.0) and neutralized by the addition of Tris-HCl (pH 9.0). The eluted material was bufferexchanged intoPBSusingNap10columns (Amersham,#17‑0854‑02)and theconcentrationof IgGwasdetermined spectrophotometrically using an extinction coefficient based on the amino acid sequence of the IgG (Mach et al., Anal. Biochem. 200(1):74‑80 (1992)). The purified IgGwere analyzed for aggregation and degradation purity using SEC-HPLC and by SDS-PAGE. Screening for pH sensitive Par0067 variant scFv’s and IgG’s

[0142] In order to screen for and profile antibodies with potential pH dependent binding a biochemical epitope competition assay format was used. The assay, which used Homogeneous Time Resolved Fluorescence (HTRF™) technology, was designed to evaluate the ability of test antibodies (scFv or IgG) to inhibit the interaction of the parent Par0067 IgGantibody binding to the humanPAR2extracellular domain (ECD). Importantly the assaywas implemented at two different pH values (pH 7.4 and pH 6.0).

[0143] Parallel assays at pH 7.4 and pH 6.0 (as described above) were first used to screen crude un-purified scFv (bacterial extracts) in a single point 384 well high throughput screen (HTS). This single point parallel HTS format enabled the screening of many 1000’s of test scFv and those antibodies which resulted in reduced inhibition of the interaction of parent Par0067 IgG binding to human PAR2 at pH 6.0 versus pH 7.4 were taken forward for further characterisation. Subsequently the same epitope competition assay approach was then implemented in a multipoint dose response IC50 format to test bothpurifiedscFvandpurified IgG (in the latter caseaminormodificationof theassaydesignwas requiredas set out in Section C). Again assays were implemented at pH 7.4 and pH 6.0 however in these experiments we were most interested in test antibodies where dose response inhibition curves showed a large rightward shift (i.e. significantly increased IC50 value) at pH 6.0 relative to corresponding dose response inhibition curves observed at pH 7.4.

[0144] The following protocol includes methods for both single point testing of crude un-purified scFv and subsequent testing of purified scFv and IgG. Section A: Generic Assay Conditions: Assay buffers:

[0145] Assay buffers were made fresh on the day of use. For experiments at pH 7.4 DPBS (Gibco 14190‑086) was supplemented with KF (0.4M) (VWR, 103444T) and BSA (0.1% w / v) (PAA, K05‑013) followed by rechecking the pH and 29 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 fineadjusting to pH7.4 as required. For experiments at pH6.0,while keepingall other buffer components identical with the pH 7.4 assay buffer outlined above, the pH 6.0 assay buffer was formulated using a base buffer of 200 mMMES (Sigma, M‑5287) (as opposed to DPBS). After addition of KF (0.4M) and BSA (0.1%w / v) the 200mMMES based buffer was then adjusted to pH 6.0 with HCL. Assay Plates: Assays were performed using black shallow well 384 plates (Round bottom, non-binding) (Corning, 4514) Assay Volume: 20 µl Incubation and Plate Reading: Assay plates were incubated for 2hrs at RT before reading using a standard HTRF™ read protocol on an Envision plate reader. Section B: Testing of scFv: (Un-purified bacterial lysates and purified scFv) Addition Order and Assay Components:

[0146] Total nsb Test Par0067 IgG (x4 [final]) 5µl 5µl 5µl Test scFv (x4 [final]) - - 5µl Assay Buffer 5µl 5µl - Bio-human-PAR2 ECD (x4 [final]) 5µl - 5µl Assay Buffer - 5µl - Europium Cryptate Labelled Streptavidin plus XL665 labelled anti-human-Fe (both x4 [final]) 5µl 5µl 5µl Par0067 IgG Preparation / Addition:

[0147] Unlabelled purified Par0067 IgG (generated in house) wasmade up to a concentration of 4.44 nM (1.11 nM final [assay]) in eachof the twoassaybuffers previously described inSectionA (pH7.4&pH6.0). 5µl / well of the appropriate pH 4.44 nM Par0067 IgG solution was added to all wells of the relevant assay plate (pH 7.4 and pH 6.0). Test scFv Preparation / Addition:

[0148] a) For parallel single point HTS of crude un-purified bacterial lysate scFv samples at both pH 7.4 and pH 6.0 samples were first pre-diluted to 40% of their neat concentration using either pH 7.4 or pH 6.0 assay buffer as appropriate. Subsequently 5µl of 40% pre-diluted sample was then transferred into the appropriate assay (pH 7.4 or pH 6.0) in order to give a final assay sample concentration of 10.0% (in the 20µl final assay volume). The parent Par0067 scFv was included inallHTSexperimentsasacontrol aswerespecificpHdependentantibodiesas thosebecameavailable (for benchmarking). b) Formultipoint dose response IC50 testing of purified scFv antibody, sampleswere tested froma top final assay of¼ of the neat undiluted sample (i.e. no pre-dilution stepwas performed). Duplicate 11 point 1:3 serial dilutions thenwere prepared on 384well polypropyleneGreiner plates in each of the two assay buffers (pH7.4& 6.0). 5µl perwell of each serial dilutionwas transferred from the relevant pHscFvdilutionplate to the correspondingassayplate (pH7.4 andpH 6.0). 5µl of the appropriate pHassay bufferwas added to the total and non-specificwells. The parent Par0067purified scFv was included in all multipoint dose response IC50 experiments as a control as were specific pH dependent purified scFv as those became available (for benchmarking). Data are presented in Table 1. Biotinylated Human PAR2 ECD Preparation / Addition:

[0149] In house generated biotinylated human PAR2 ECD was diluted into each of the two assay buffers (pH 7.4 & pH 6.0) to give working solutions at 4.0 nM (1 nM final assay concentration). 5µl / well of the appropriate 4.0 nM biotinylated human PAR2ECDworking solution was then added to all wells of the corresponding assay plate (pH 7.4 and pH 6.0) with the exception of the negative binding control wells. 5µl / well of the appropriate pHassay bufferwas added to the negative 30 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 binding wells. HTRF Detection Reagent Preparation / Addition:

[0150] Europium Cryptate Labelled Streptavidin (CisBio, 610SAKLB) and XL665 Labelled Anti-Human-Fc (CisBio, 61HFCXLB) were each diluted into each pH assay buffer (pH 7.4, & pH 6.0) to give combined working solutions at concentrationsof 6.0nM (EuropiumCryptate LabelledStreptavidin) and40nM (XL665 LabelledAnti-Human-Fc). Allowing for the x4 fold dilution into the assay this resulted in final assay concentrations of 1.5 nM (Europium Cryptate Labelled Streptavidin) and10nM (XL665 LabelledAnti-Human-Fc). 5µl / well of the relevant pHcombinedHTRF™detection reagent working solution was then added to all wells of the corresponding assay plate (pH 7.4 and pH 6.0). Section C: Testing of Purified IgG: Addition Order and Assay Components:

[0151] Total nsb Test Dylight650 labelled Par0067 IgG (x4 [final]) 5µl 5µl 5µl Test IgG (x4 [final]) - - 5µl Assay Buffer 5µl 5µl - Bio-human-PAR2 ECD (x4 [final]) 5µl - 5µl Assay Buffer - 5µl - Europium Cryptate Labelled Streptavidin (x4 [final]) 5µl 5µl 5µl Par0067 IgG Dylight650 Labelling:

[0152] Dylight650 labelling of Par0067 IgG was performed using a Dylight650 labelling kit (Thermo Scientific Cat. No. 84536). Labelling of in-house generated purified Par0067 IgG was performed according to the manufacturer’s recom- mended labellingprocedure.ThefinalDylight650 labelledPar0067 IgGconcentrationwasdeterminedas0.56mg / mlwitha mean Dylight650 to IgG dye incorporation ratio of 2.7 moles dye / mole IgG. Dylight650 Labelled Par0067 IgG Preparation / Addition:

[0153] Dylight650 labelled Par0067 IgG (0.56mg / ml, 3,733 nM) wasmade up to a concentration of 4.44 nM (to give 1.11 nMfinal [assay]) in eachof the twoassaybufferspreviously described in thematerials section (pH7.4&pH6.0). 5µl / well of the appropriate pH 4.44 nMDylight650 Par0067 IgG solution was added to all wells of the relevant assay plate (pH 7.4 and pH 6.0). Test IgG Serial Dilution / Addition:

[0154] ParentPar0067 IgG (usedasa reference / control in all assays)waspre-diluted to givea2000nMstock ineachof the twodifferent pHassaybuffers (in order togivea top final assay IgGconcentrationof 500nM).All test, or other reference / control IgGwere tested froma topfinal assay concentrationof¼of theneatundilutedsample (i.e. nopre-dilution stepwas performed). Duplicate 11 point 1:3 serial dilutions thenwere prepared on 384well polypropyleneGreiner plates in each of the two assay buffers (pH 7.4 & 6.0). 5µl per well was transferred from the relevant pH IgG dilution plate to the corresponding assay plate (pH 7.4 and pH 6.0). 5ul of the appropriate pH assay buffer was added to the total and non-specific wells. Biotinylated Human PAR2 ECD Preparation / Addition:

[0155] In house generated biotinylated human PAR2 ECD was diluted into each of the two assay buffers (pH 7.4 & pH 6.0) to give working solutions at 4.0 nM (1 nM final assay concentration). 5µl / well of the appropriate 4.0 nM biotinylated human PAR2ECDworking solution was then added to all wells of the corresponding assay plate (pH 7.4 and pH 6.0) with the exception of the negative binding control wells. 5µl / well of the appropriate pHassay bufferwas added to the negative binding wells. 31 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 HTRF Detection Reagent Preparation / Addition:

[0156] Europium Cryptate Labelled Streptavidin (CisBio, 610SAKLB) was diluted into each pH assay buffer (pH 7.4, & pH 6.0) to give work solutions at a concentration of 6.0 nM. Allowing for the x4 fold dilution into the assay this resulted in a final assay Europium Cryptate Labelled Streptavidin concentration of 1.5 nM. 5µl / well of the relevant pH Europium Cryptate Labelled Streptavidin working solution was then added to all wells of the corresponding assay plate (pH 7.4 and pH 6.0). Section D: Data Analysis:

[0157] 665nmand620nmcountswerefirst converted to665 / 620 ratio values.DeltaF (%)was thencalculatedaccording to the following equation: Negative ratio values were calculated in the absence of Par0067 IgG from the relevant negative binding control wells.

[0158] % Specific Binding was then calculated according to the following equation: % Specific Binding ={ (Sample % Delta F - Negative Binding % Delta F) / (Total Binding % Delta F - Negative Binding % Delta F)} * 100

[0159] For singlepointHTS%SpecificBindingat pH6.0versuspH7.4wasplottedonxandyaxes, respectively, in order to visualise the distribution of scFv with reduced inhibition (higher % Specific Binding) at pH 6.0 relative to pH 7.4.

[0160] For multipoint dose response curves % Specific Binding values were plotted versus test purified antibody concentration (scFv or IgG). IC50 valueswere determined via a sigmoidal dose response inhibition variable slope curve fit (4 parameter logistic equation) using Graphpad Prism Software. TABLE 1 Clone (scFv) Par0067 Epitope Competition Assay pH7.4 IC50 (nM) pH6.0 IC50 (nM) Variant : Fold IC50 (pH6.0 v pH7.4) Fold IC50 pH7.4 (Variant v Par0067) Par0067 3.3 3.6 1.1 1.0 PaB670010 15.4 41.8 2.7 4.7 PaB670020 5.6 18.5 3.3 1.7 PaB670034 5.6 14.2 2.5 1.7 PaB670045 9.8 72.5 7.4 3.0 PaB670048 14.5 139.4 9.6 4.4 PaB670064 67.1 289.9 4.3 20.3 PaB670066 52.2 486.9 9.3 15.8 PaB670067 14.4 113.2 7.9 4.4 PaB670068 77.8 447.6 5.8 23.6 PaB670070 60.4 270.0 4.5 18.3 PaB670071 101.2 424.8 4.2 30.7 PaB670073 72.1 428.3 5.9 21.8 PaB670075 101.8 603.0 5.9 30.8 PaB670076 46.2 291.8 6.3 14.0 PaB670077 55.4 619.9 11.2 16.8 PaB670078 33.3 319.2 9.6 10.1 PaB670079 44.2 489.8 11.1 13.4 32 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) Clone (scFv) Par0067 Epitope Competition Assay pH7.4 IC50 (nM) pH6.0 IC50 (nM) Variant : Fold IC50 (pH6.0 v pH7.4) Fold IC50 pH7.4 (Variant v Par0067) PaB670080 16.6 158.1 9.5 5.0 PaB670081 51.4 260.0 5.1 15.6 PaB670082 25.4 77.9 3.1 7.7 PaB670083 45.2 151.8 3.4 13.7 PaB670084 54.5 282.7 5.2 16.5 PaB670085 48.1 154.7 3.2 14.6 PaB670087 40.9 130.7 3.2 12.4 PaB670088 29.8 114.7 3.8 9.0 PaB670089 29.9 134.0 4.5 9.1 PaB670090 22.6 85.5 3.8 6.8 PaB670091 25.2 98.9 3.9 7.6 PaB670092 41.1 193.5 4.7 12.5 PaB670093 18.6 58.7 3.2 5.6 PaB670094 72.5 236.8 3.3 22.0 PaB670095 20.0 113.2 5.7 6.1 PaB670097 21.8 77.9 3.6 6.6 PaB670098 65.9 210.3 3.2 20.0 PaB670099 12.9 62.4 4.8 3.9 PaB670100 20.3 69.0 3.4 6.2 PaB670101 146.9 496.5 3.4 44.5 PaB670102 5.3 13.7 2.6 1.6 PaB670103 27.9 203.5 7.3 8.5 PaB670104 21.7 202.2 9.3 6.6 PaB670105 74.4 495.5 6.7 22.5 PaB670106 312.8 1188.0 3.8 94.8 PaB670107 42.7 463.9 10.9 12.9 PaB670108 29.1 148.6 5.1 8.8 Recombination of multiple histidines into a single scFv

[0161] Histidine residues from scFv which demonstrated pH dependent binding in the Par0067 epitope competition binding assay were recombined using site directed mutagenesis (Reikofski J and Tao BY, 1992, Biotechnol Adv, 10(4): 535‑547). Resulting scFvwhich contained histidines in two different CDRswere retested in the competition binding assay as whole immunoglobulin G1 triple mutant (IgG1-TM) (Table 2) to identify variants with further improvements in pH dependent binding compared to the parent antibody, Par0067.

[0162] Sequence alignments for each of the histidine modified clones as compared to the reference Par0067 antibody CDR sequences are provided in Figures 1A‑1B and 2A‑2B. 33 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 TABLE 2 Clone (IgG) Par0067 Epitope Competition Assay pH7.4 IC50 (nm) pH6.0 IC50 (nM) Variant : Fold IC50 (pH6.0 v pH7.4) Fold IC50 pH7.4 (Variant v Par0067) Par0067 0.835 0.790 0.95 1.00 PaB670045 2.9 25.8 8.9 3.5 PaB670048 5.7 48.2 8.5 6.8 PaB670128 32.8 IC ND 39.3 PaB670129 56.1 IC ND 67.2 PaB670084 10.3 64.6 6.3 12.3 PaB670141 2.4 13.9 5.9 2.8 PaB670142 4.8 67.1 14.0 5.7 PaB670143 3.4 40.0 11.8 4.0 PaB670144 29.0 564.8 19.5 34.7 PaB670146 91.9 784.3 8.5 110.1 PaB670148 48.5 844.6 17.4 58.1 PaB670149 504.9 IC ND 604.7 PaB670151 69.5 796.0 11.5 83.2 PaB670152 438.8 6604.0 15.1 525.5 PaB670153 93.0 1639.0 17.6 111.4 PaB670156 7.0 190.0 27.2 8.4 PaB670157 15.8 932.4 59.0 18.9 PaB670158 6.8 477.0 69.8 8.2 PaB670159 161.0 IC ND 192.8 PaB670160 16.9 634.3 37.5 20.2 PaB670161 47.9 1899.0 39.6 57.4 PaB670162 11.4 906.2 79.5 13.7 PaB670163 145.1 IC ND 173.8 IC = Incomplete Curve; ND = Not Demonstrated Affinity of anti-PAR2 Fabs for human, rat and cynomolgus PAR2 as determined by BIACORE™

[0163] Antigen binding fragments (Fabs) of the anti-PAR2 antibodies were expressed (Spooner J. et al. (2015) Biotechnol Bioeng. 112:1472‑7) and the affinity for recombinant PAR2 of various species (human, rat and cynomolgus) determined by Biacore. Biacore Affinity Analysis

[0164] The affinity of the anti-PAR2 Fabswasmeasured using the Biacore T100 at 25°C at various pH’s (pH 7.4, pH 6.0 and pH 5.6). The experiments were carried out using recombinant human, rat and cynomolgus PAR2 with an N-terminal Avi tag and C-terminal Flag-His tags.

[0165] Streptavidin was covalently immobilised to a C1 chip surface using standard amine coupling techniques at a concentration of 4 µg / ml in 10 mMSodium acetate pH 4.5. A final streptavidin surface of approximately 30‑100 RUs was achieved. Recombinant biotinylated PAR2 species (produced in-house) were titrated onto the streptavidin chip surface at 4 µg / ml in HBS-EP+ buffer to enable Fab binding at saturation (Rmax). This low level of analyte binding ensured minimal mass transport effects. 34 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55

[0166] The anti-PAR2 Fabs were serially diluted (0.39 nM ‑25 nM) in HBS-EP+ buffer pH 7.4 or MES-BS-EP+ pH 6.0 buffer or in MES-BS-EP+ pH 5.6 buffer and flowed over the chip at 50 µl / min, with 3 minutes association and up to 30 minutes dissociation. Multiple buffer-only injections were made under the same conditions to allow for double reference subtraction of the final sensorgramsets,whichwereanalysedusing theBiaEval software (version 2.0.1). The chip surface was fully regenerated with pulses of 4 M MgCl2.

[0167] BiaCore affinity results for select clones are provided below in Tables 3‑13. Table 3: Par0067 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 6.66 E+6 4.96 E‑5 7.5 50.1 6.0 Human 3.12 E+6 9.72 E‑5 31.2 39.3 7.4 Rat 5.16 E+6 1.94 E‑4 37.6 111 6.0 Rat 3.21 E+6 5.44 E‑4 170 100 Table 4: PaB670048 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 4.20 E+6 5.19 E‑4 124 69.0 6.0 Human 1.94 E+6 4.97 E‑3 2569 58.3 7.4 Rat 5.24 E+6 3.07 E‑3 586 124.7 6.0 Rat 4.00 E+6 2.05 E‑2 5120 105.2 Table 5: PaB670084 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 6.49 E+6 1.62 E‑3 250 68.4 6.0 Human 1.73 E+6 7.63 E‑3 4,410 104.6 7.4 Rat 6.83 E+6 8.37 E‑3 1226 97.6 6.0 Rat 2.57 E+6 2.77 E‑2 10,800 91.97 Table 6: PaB670076 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 5.84 E+6 1.21 E‑3 207 63.5 6.0 Human 1.11 E+6 5.14 E‑3 4,611 99.51 7.4 Rat 6.34 E+6 1.07 E‑2 1,689 87.9 6.0 Rat 1.66 E+6 5.10 E‑2 30,730 82.61 Table 7: PaB670120 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 5.54 E+6 1.37 E‑3 247 61.62 6.0 Human 3.48 E+6 2.10 E‑2 6,047 94.67 7.4 Rat 5.62 E+6 1.46 E‑2 2,606 84.23 6.0 Rat 8.26 E+6 0.281 34,040 73.35 35 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 Table 8: PaB670128 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 5.95 E+6 3.65 E‑3 613 59.57 6.0 Human 2.22 E+6 4.57 E‑2 20,062 79.4 7.4 Rat 6.49 E+6 1.96 E‑2 3,023 80.86 6.0 Rat 1.79 E+6 6.89 E‑2 38,520 63.26 Table 9: PaB670048 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 5.59 E+6 5.80 E‑4 104 58.53 6.0 Human 1.84 E+6 4.58 E‑3 2,488 93.02 7.4 Rat 7.46 E+6 3.25 E‑3 435 81.2 6.0 Rat 4.84 E+6 2.17 E‑2 4,483 80.14 Table 10: PaB670129 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 8.20 E+6 5.04 E‑3 614 54.98 6.0 Human 1.59 E+6 5.54 E‑2 34,840 76.02 7.4 Rat 8.82 E+6 2.63 E‑2 2,979 73.16 6.0 Rat 1.45 E+6 8.41 E‑2 57,910 67.27 Table 11: PaB670136 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 5.17 E+6 4.95 E‑3 957 50.27 6.0 Human 7.92 E+8 14.9 18,840 67.97 7.4 Rat 7.57 E+6 3.14 E‑2 4,149 73.24 6.0 Rat 1.57 E+6 5.89 E‑2 37,550 47.7 Table 12: PaB670103 Fab. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 3.52 E+6 1.11 E‑4 32.5 46.3 6.0 Human 3.36 E+5 6.07 E‑4 1,805 70.66 7.4 Rat 3.04 E+6 5.58 E‑4 184 74.2 6.0 Rat 5.31 E+5 5.42 E‑3 10,200 69.58 Table 13: PaB670129 Fab at 3 different pH’s. pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Human 7.14 E+6 5.34 E‑3 747.8 75.9 6.81 E+6 5.30 E‑3 778.0 66.5 6.0 Human 1.56 E+6 5.76 E‑2 36,910 58.1 3.26 E+6 0.109 33,500 48.4 5.6 Human 1.33 E+6 0.112 84,550 42.5 36 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) pH Species ka (M‑1 s‑1) kd (s‑1) KD (pM) Rmax 7.4 Cynomolgus 6.67 E+6 2.23 E‑3 333.3 59.7 6.59 E+6 2.21 E‑3 335.0 51.9 6.0 Cynomolgus 2.14 E+6 2.75 E‑2 12,840 49.6 1.74 E+6 2.36 E‑2 13,560 44.2 5.6 Cynomolgus 3.66 E+6 0.139 37,840 41.3 Example 2: Cell-Based PAR2 and PAR1 Activity Assay

[0168] Human A549 cells, rat KNRK, mouse LL / 2 or cynomolgus CYNOM-K1 cells expressing endogenous PAR2, or human1321N1-hPAR2-c18cells overexpressinghumanPAR2wereseededat 5,000 (human, cyno) or 7,000 (mouse, rat) cells perwell onPDL-coatedTissueCulturePlates, (GreinerBio-One).Cellswere loadedwithFluo-ScreenQuest™Fluo‑8 NoWash Calcium dye (AAT Bioquest, Inc). Cells were pre-treated with IgGs or Fabs diluted in assay buffer (HBSS, 0.1% BSA,20mMHEPES) for 1hat room temperature. Formouseandcynoassays, cellswerepre-treatedwith0.5nMor10nM thrombin, respectively, to desensitizePAR1activity. PAR2calcium responses to11nM (human), 400nM (mouse) or 80nM (rat, cyno) trypsin (Polymun) were measured on a Fluorescent Imaging Plate Reader (FLIPR) Tetra (Molecular Devices). Todetermine functional activity against humanPAR1, thrombindriven calcium responses in theA549humancell linewere determined also on the FLIPR tetra and in these assays neutralising anti-PAR1 IgGs WEDE15 (Beckman Coulter) and ATAP2 (Life Technologies) were used as positive controls. To determine functional activity against diverse proteases, 1321N1-hPAR2-cl8 cells overexpressing human PAR2 were pre-treated with PaB670129, prior to stimulating calcium release with 0.5 nM trypsin, 500 nM tryptase or 1 nM matriptase. Fluorescence measurements were measured before, during, and after protease addition and peak RFU calculated per well. % responses (relative to protease alone) were calculated against antibody concentration and IC50s determined using GraphPad Prism software.

[0169] Results fromhuman, cynomologus-monkey, rat, andmousecells in theseassaysareprovided inFigure 3 (A,B,C and D respectively), calculated PAR2 IC50s in Figure 3E and Figure 6 (PAR1 specificity data). IC50 calculations demonstrate that PaB670129 potently inhibits trypsin-induced PAR2 calcium responses in human, mouse, rat and cyno cells expressing endogenous PAR2 (Figure 3E). Furthermore in human A549 cells, thrombininduced PAR1 activation is not inhibited byPaB670129but can beeffectively blockedby theVorapaxar and anti-PAR1monoclonal antibodies (Figure 6). These data demonstrate that PaB670129 is a potent and specific antagonist of PAR2. Application of PaB670129 alone toPAR2-expressingcells hasnoeffect onbasal cellular calcium levels, demonstrating thatPaB670129 lacksanyagonistic activity at PAR2 (Figure 4A). Furthermore, PaB670129 potently antagonizes PAR2-evoked responses to diverse proteases including trypsin, tryptase and matriptase (Figure 4B). Primary DRG Glial-neuronal PAR2 Calcium Assay

[0170] Dissociated cultures of Sprague dawley rat pup dorsal root ganglia (DRG) were prepared and grown on laminin andPDL-coatedTissueCulturePlates (GreinerBio-One). Plateswere incubatedat 37degrees for 24‑72hoursbefore use in the assay. Cells were loaded with 2 µMFura‑2 calcium dye (Life Technologies). Cells were incubated in imaging buffer (HBSS, 20 mM HEPES, 0.1 mM Sulfinpyrazone, 10 µM PAR1 antagonist Vorapaxar) containing PAR2 antibodies at 20 nM. Intracellular calciumwas thenquantifiedbyFura‑2 ratiometric imagingonanOlympus IX81microscopeequippedwith a Xenon arc lamp exciting at 340 and 380 nm in response to application of the agonists thrombin (Sigma) and matriptase (R&D Systems) PAR2 activating peptide LIGRLO (SEQ ID NO: 832), (Peptides International) and high extracellular potassium (50 mM). Number of neurones versus glia was calculated per field of view (neurones defined as showing response to high potassium). Total matriptase-sensitive neurones and glia were then calculated per field of view. Results from the calcium assay are provided in Figures 5A-F and illustrate that the PaB670129 antibody effectively reduced sensitivity to matriptase in DRG neuronal (Figure 5A-C) and non-neuronal cells (Figure 5D-F). Example 3: Effects of an Anti-PAR2 Antibody in a Rat Model of inflammatory joint Pain

[0171] Intra-articular administration of Monosodium Iodoacetate (MIA) in the ipsilateral knee of Sprague Dawley rats leads to development of a robust and long-lasting hyperalgesia and allodynia associated initially with an inflammatory response. The development of these signs in this animal model are believed to be clinically relevant; reflecting the symptoms displayed by patients presentingwith chronic inflammatory pain associatedwith underlying conditions such as osteoarthritis (OA)or rheumatoidarthritis (Boveetal., 2003;Fernihoughetal., 2004;Kalbhen1987). It haspreviouslybeen 37 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 demonstrated that (usingweight-bearingasanend-point) the time courseofMIA inducedhyperalgesia followsabi-phasic patternwith an early, predominantly inflammatory, componentwhich isCox‑2 sensitive; andmarkedly reduced by the gold standardCelecoxib. Thisearly inflammatoryphasegivesway toamorechronicpainphenotypewhich isPregabalin (PGB) sensitive and Celecoxib insensitive suggesting an underlying more neuropathic-type component.

[0172] Weight bearing: Naive rats distribute their body weight equally between the two hind paws. However, when the injected (left) hind knee is inflamed and / or painful, theweight is re-distributed so that lessweight is put on the affected limb (decrease inweight bearing on injured limb).Weight bearing througheachhind limb ismeasuredusing a rat incapacitance tester (Linton Instruments, UK).

[0173] Sprague Dawley rats were placed in the incapacitance tester with the hind paws on separate sensors and the average force exerted by both hind limbs was recorded over 4 seconds.

[0174] Procedure:After delivery, rats underwent aminimumhabituation period of 7 days prior to study commencement. Naïve rats were acclimatised to the procedure room in their home cages, with food and water available ad libitum. Habituations to the weight bearing chamber were performed over several days. Base line weight bearing readings were taken on the final day.

[0175] On Day 0, following the final baseline reading, animals were anaesthetised using isoflurane and oxygen mixed 3:1 in sterile conditions. The left knee area was shaved and cleaned with a dilute hibiscrub solution.

[0176] Osteoarthritis (OA) was induced via injection of MIA (Sigma, 12512) solution, 25 µl of 80 mg / ml, (2 mg) into the knee joint of the left hind leg. Sham animals were injected with saline. Animals were allowed to recover in a warmed environment, before being returned to their home cage.

[0177] Animals develop an inflammatory response post MIA and may guard and lick the affected area. Rats were therefore carefully monitored for unexpected signs of distress or severe pain, so that any animal displaying such signs could be culled immediately.

[0178] Animals were weighed daily for the first week and then every few days after. Weight bearing was assessed on Days 3, 7, 10 & 14, following injection of MIA, for development of chronic pain. On day 18, weight bearing measurements were taken and animalswere ranked and randomised to treatment groups according to theirMIAwindow in a Latin square design.

[0179] Antibody dosing regimen:Animals were treated with Par0067 (PAR2+ve) 10mg / kg or isotype control (PAR2-ve) 10mg / kg i.v. on day 18 and further weight bearing measurements were taken 4 hours and 1, 2, 6, 8, 10 & 14 days post antibody dosing.

[0180] Pregabalin & Celecoxib dosing regimen: Animals were dosed with PGB (30 mg / kg p.o; 2 ml / kg) or Celecoxib (50mg / kgp.o; 2ml / kg) daily ondays 24, 25, 26, 27 and28 postMIA injection.Weight bearing assessmentswere taken1hr post dosing on days 24, 26 and 28 and a further reading was taken post cessation of drug treatment on day 32.

[0181] Onday 1, 2, 6, 10 and 14 post dosing, after weight bearing assessment, 400µl of bloodwas taken via the tail vein for pk analysis from Antibody and Isotype control treated groups (n=5 / group).

[0182] Evaluation of Study:Weight bearing (g) readings were taken for both right and left hind paws and the difference calculated. Data are expressed as % ratio ipsilateral / contralateral ((WB left / WB right)* 100) (mean ± s.e.m.).

[0183] Calculation: Ipsilateral reading / contralateral reading x 100. Naïve WB difference - pre dose WB difference was defined as the MIA window.

[0184] Statistical analysis: Repeated measures ANOVA followed by Planned comparison test using InVivoStat (invivostat.co.uk), (p<0.05 considered significant). Data were analysed by comparing treatment groups to vehicle control group at each time point.

[0185] Injection of 2 mgMIA into the knee joint caused amarked inflammation and hypersensitivity response apparent from day 3 as detected by a shift in weight bearing between injured and non-injured hind paws. This MIA-induced hyperactivity response was still evident in all groups up until day 18 (and beyond for vehicle-treated control animals) at which point the first test agents were administered. Injection of saline had no effect on weight bearing.

[0186] As demonstrated in Figure 7A, a significant and marked reversal of hypersensitivity was seen after daily administration of Pregabalin (30mg / kg) from day 24‑28 with a weak residual effect still apparent after prior cessation of treatment on day 32. In contrast, daily administration of Celecoxib (50 mg / kg) from day 24‑28 showed only a weak reversal of hypersensitivity at best. This pharmacological profile suggests that the hyperactivity observed during this phase of the MIA response is predominantly neuropathic, rather than inflammatory, in nature.

[0187] As also demonstrated in Figure 7A, a significant reversal of hypersensitivity was seenwith Par0067 from4 hours post dose, through until day 28 (10 days post dose). No effect was seen with Isotype Control during the same time period. Figure 7B illustrates the effect of treatment with different concentrations of Par0067. Example 4: The Effect of the PAR2 antibody, PaB670129 on Reversal of Partial Nerve Ligation - Induced Mechanical Hyperalgesia in Female C57BL / 6 Mice 38 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 Introduction

[0188] Partial ligation of the sciatic nerve (PNL) as described by Seltzer (1990) is one of a number of nerve ligation models which are reported to serve as pre-clinical models of neuropathic pain. It produces a profound mechanical hyperalgesia which can be measured using an analgysemeter as described by Randall and Selitto (1957). This example describes the effects of the administration of the anti PAR2 antibody PaB670129, on hyperalgesia in this nerve injury / neuropathic pain model. Procedure

[0189] Sixty femaleC57BL / 6mice underwent insertion of transponders for identification purposesat least 5 days before the start of the study. Mechanical hyperalgesia was determined using an analgysemeter (Randall & Selitto 1957) (Ugo Basile). An increasing force was applied to the dorsal surface of each hind paw in turn until a withdrawal response was observed. The application of force was halted at this point and the weight in grams recorded. Data was expressed as withdrawal threshold in grams for ipsilateral and contralateral paws. Following the establishment of baseline readings, mice were divided into 2 groups with approximately equal ipsilateral / contralateral ratios which underwent surgery to partially ligate the sciatic nerve or served as sham operated controls. Operated mice were anaesthetised with isoflurane. Following this, approximately 1 cmof the left sciatic nervewas exposed by blunt dissection through an incision at the level of themid thigh.Asuture (8 / 0VirginSilk:Ethicon)was thenpassed through thedorsal third of thenerveand tied tightly. The incision was then closed using glue and the mice were allowed to recover for at least six days prior to commencement of testing.Shamoperatedmiceunderwent thesameprotocol, but followingexposureof thenerve, themiceweresuturedand allowed to recover.

[0190] Mice were tested for onset of hyperalgesia on days 7 and 10 post surgery. Any mice showing an ipsilateral / con- tralateral ratio of greater than 80%were classed as non-responders and removed from the study. Following testing on day 10, mice were further sub-divided into groups giving the final treatment groups; A. B. C. D. E. F.

[0191] Mice were administered control or test molecules diluted in Phosphate Buffered Saline, (PBS) on day 13 and were re-tested for changes in mechanical hyperalgesia at 4 hours post dose and also on 1, 2, 4 and 7 days post dose. Data Analysis

[0192] Ipsilateral and contralateral readings were taken for each animal at each test time. Weight bearing through ipsilateral and contralateral hind limbs was expressed as a ratio and the group data were analysed (PRISM) using 2-way 39 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 ANOVA and pairwise comparisons where appropriate were made using Tukey’s test. Results

[0193] Partial ligationof the sciatic nerve causedamechanical hyperalgesiawhichmanifestedasa significant reduction in the ipsilateral / contralateral ratio on day 7 and 10 when compared to sham operated controls. Following treatment with isotype control, operated mice did not show any change in the level of mechanical hyperalgesia from pre-dose levels indicating a lack of effect. The administration of the internal gold standard, etanercept (0.3mg / kg s.c.) caused a significant reversal of the hyperalgesia from 4 hours through to 7 days post dose in agreement with the results seen in previous studies. PaB670129, caused a reversal of the ipsilateral / contralateral ratio in a dose related fashion with peak effects beingseenatboth10mg / kgand50mg / kg.The lowest doseof3mg / kgwhilst significantat 1,2and7dayspostdoseshowed a smaller effect (see Figure 8).

[0194] Partial ligation of the sciatic nerve induced a long lasting mechanical hyperalgesia consistent with previously reported results. Without wishing to be bound by theory, this is believed to serve as a pre-clinical correlate of the pain observed in neuropathic pain. The administration of PaB670129, showed a significant and dose related reversal of this hyperalgesia indicating a potential use of PAR2 antibodies in the treatment of neuropathic pain. Bibliography:

[0195] • Persic, L. et al. An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries. Gene 187, 9‑18 (1997). • Reikofski J and Tao BY (1992) Polymerase chain reaction (PCR) techniques for site-directed mutagenesis. Biotechnol Adv, 10(4): 535‑547. • Bove SE, Calcaterra SL, Brooker RM, Huber CM, Guzman RE, Juneau PL, et al. Weight bearing as a measure of disease progression and efficacy of anti-inflammatory compounds in a model of monosodium iodoacetate-induced osteoarthritis. Osteoarthritis Cartilage 2003; 11 (11): 821‑30. eng. • Fernihough J, Gentry C, Malcangio M, Fox A, Rediske J, Pellas T, et al. Pain related behaviour in two models of osteoarthritis in the rat knee. Pain 2004; 112 (1‑2): 83‑93. eng. • KalbhenDA.Chemicalmodel of osteoarthritis--a pharmacological evaluation. JRheumatol 1987; 14SpecNo: 130‑1. eng. • Clark RA, Shoaib M, Hewitt KN, Stanford SC, Bate ST (2012)., A comparison of InVivoStat with other statistical software packages for analysis of data generated from animal experiments, J Psychopharmacology, 26(8), 1136‑1142. • Myska Improving Biosensor Analysis. Journal of Molecular Recognition. 1999 • D.G. Myska Improving Biosensor Analysis. Journal of Molecular Recognition. 1999;12 : 279‑284. • Myska DG, Improving Biosensor Analysis. Journal of Molecular Recognition. 1999; 12: 279‑284. • A.W. Drake, M.L. Tang, G.A. Papalia, G. Landes, M. Haak-Frendscho, S.L. Klakamp, Biacore surface matrix effects on the binding kinetics and affinity of an antigen / antibody complex, Anal. Biochem. 429 (2012) 58‑69 • Pace CN, Vajdos F, Fee L, Grisley G and Grey T, How to measure and predict the molar absorption coefficient of a protein, Protein Sci. 1995; 4: 2411‑2423. • Spooner J, Keen J, Nayyar K, Birkett N, Bond N, Bannister D, Tigue N, Higazi D, Kemp B, Vaughan T, Kippen A, Buchanan A. (2015) Evaluation of strategies to control Fab light chain dimer during mammalian expression and purification: A universal one-step process for purification of correctly assembledFab. Biotechnol Bioeng. 112:1472‑7. • Daramola O, Stevenson J, Dean G, Hatton D, Pettman G, Holmes W, Field R (2014) A high yielding CHO transient 40 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 system: co-expression of genes encoding EBNA‑1 and GS enhances transient protein expression. Biotechnol Prog. 30(1):132‑41. • Mach H, Middaugh CR, Lewis RV (1992) Statistical determination of the average values of the extinction coefficients of tryptophan and tyrosine in native proteins. Anal. Biochem. 200(1):74‑80. • SeltzerZ,DubnerR,ShirY (1990).Anovel behaviouralmodel of neuropathic paindisordersproduced in ratsbypartial sciatic nerve injury. Pain 43: 205‑218 • Randall LO, Selitto JJ (1957). A method for measurement of analgesic activity on inflamed tissue. Arch Int Pharmacodyn Ther. 111(4): 409‑419 SEQUENCE LISTING:

[0196] PRT= amino acid sequence SEQ ID NO: clone name Type 1 ZZ15D2-D02 (Par0067) VH DNA 2 ZZ15D2-D02 (Par0067) VH PRT 3 ZZ15D2-D02 (Par0067) CDR1 PRT 4 ZZ15D2-D02 (Par0067) CDR2 PRT 5 ZZ15D2-D02 (Par0067) CDR3 PRT 6 ZZ15D2-D02 (Par0067) VL DNA 7 ZZ15D2-D02 (Par0067) VL PRT 8 ZZ15D2-D02 (Par0067) CDR1 PRT 9 ZZ15D2-D02 (Par0067) CDR2 PRT 10 ZZ15D2-D02 (Par0067) CDR3 PRT 11 ZZ1RUE-F02 (PaB670129) VH DNA 12 ZZ1RUE-F02 (PaB670129) VH PRT 13 ZZ1RUE-F02 (PaB670129) CDR1 PRT 14 ZZ1RUE-F02 (PaB670129) CDR2 PRT 15 ZZ1RUE-F02 (PaB670129) CDR3 PRT 16 ZZ1RUE-F02 (PaB670129) VL DNA 17 ZZ1RUE-F02 (PaB670129) VL PRT 18 ZZ1RUE-F02 (PaB670129) CDR1 PRT 19 ZZ1RUE-F02 (PaB670129) CDR2 PRT 20 ZZ1RUE-F02 (PaB670129) CDR3 PRT 21 ZZ1DRB-B08 (PaB670010) VH DNA 22 ZZ1DRB-B08 (PaB670010) VH PRT 23 ZZ1DRB-B08 (PaB670010) CDR1 PRT 24 ZZ1DRB-B08 (PaB670010) CDR2 PRT 25 ZZ1DRB-B08 (PaB670010) CDR3 PRT 26 ZZ1DRB-B08 (PaB670010) VL DNA 27 ZZ1DRB-B08 (PaB670010) VL PRT 28 ZZ1DRB-B08 (PaB670010) CDR1 PRT 29 ZZ1DRB-B08 (PaB670010) CDR2 PRT 30 ZZ1DRB-B08 (PaB670010) CDR3 PRT 31 ZZ1IGD-D05 (PaB670020) VH DNA 32 ZZ1IGD-D05 (PaB670020) VH PRT 33 ZZ1IGD-D05 (PaB670020) CDR1 PRT 34 ZZ1IGD-D05 (PaB670020) CDR2 PRT 35 ZZ1IGD-D05 (PaB670020) CDR3 PRT 36 ZZ1IGD-D05 (PaB670020) VL DNA 37 ZZ1IGD-D05 (PaB670020) VL PRT 41 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 38 ZZ1IGD-D05 (PaB670020) CDR1 PRT 39 ZZ1IGD-D05 (PaB670020) CDR2 PRT 40 ZZ1IGD-D05 (PaB670020) CDR3 PRT 41 ZZ1IGF-B11 (PaB670034) VH DNA 42 ZZ1IGF-B11 (PaB670034) VH PRT 43 ZZ1IGF-B11 (PaB670034) CDR1 PRT 44 ZZ1IGF-B11 (PaB670034) CDR2 PRT 45 ZZ1IGF-B11 (PaB670034) CDR3 PRT 46 ZZ1IGF-B11 (PaB670034) VL DNA 47 ZZ1IGF-B11 (PaB670034) VL PRT 48 ZZ1IGF-B11 (PaB670034) CDR1 PRT 49 ZZ1IGF-B11 (PaB670034) CDR2 PRT 50 ZZ1IGF-B11 (PaB670034) CDR3 PRT 51 ZZ1KX3-F01 (PaB670045) VH DNA 52 ZZ1KX3-F01 (PaB670045) VH PRT 53 ZZ1KX3-F01 (PaB670045) CDR1 PRT 54 ZZ1KX3-F01 (PaB670045) CDR2 PRT 55 ZZ1KX3-F01 (PaB670045) CDR3 PRT 56 ZZ1KX3-F01 (PaB670045) VL DNA 57 ZZ1KX3-F01 (PaB670045) VL PRT 58 ZZ1KX3-F01 (PaB670045) CDR1 PRT 59 ZZ1KX3-F01 (PaB670045) CDR2 PRT 60 ZZ1KX3-F01 (PaB670045) CDR3 PRT 61 ZZ1KX4-E11 (PaB670048) VH DNA 62 ZZ1KX4-E11 (PaB670048) VH PRT 63 ZZ1KX4-E11 (PaB670048) CDR1 PRT 64 ZZ1KX4-E11 (PaB670048) CDR2 PRT 65 ZZ1KX4-E11 (PaB670048) CDR3 PRT 66 ZZ1KX4-E11 (PaB670048) VL DNA 67 ZZ1KX4-E11 (PaB670048) VL PRT 68 ZZ1KX4-E11 (PaB670048) CDR1 PRT 69 ZZ1KX4-E11 (PaB670048) CDR2 PRT 70 ZZ1KX4-E11 (PaB670048) CDR3 PRT 71 ZZ1KX6-B09 (PaB670064) VH DNA 72 ZZ1KX6-B09 (PaB670064) VH PRT 73 ZZ1KX6-B09 (PaB670064) CDR1 PRT 74 ZZ1KX6-B09 (PaB670064) CDR2 PRT 75 ZZ1KX6-B09 (PaB670064) CDR3 PRT 76 ZZ1KX6-B09 (PaB670064) VL DNA 77 ZZ1KX6-B09 (PaB670064) VL PRT 78 ZZ1KX6-B09 (PaB670064) CDR1 PRT 79 ZZ1KX6-B09 (PaB670064) CDR2 PRT 80 ZZ1KX6-B09 (PaB670064) CDR3 PRT 81 ZZ1KX6-D05 (PaB670066) VH DNA 82 ZZ1KX6-D05 (PaB670066) VH PRT 83 ZZ1KX6-D05 (PaB670066) CDR1 PRT 84 ZZ1KX6-D05 (PaB670066) CDR2 PRT 85 ZZ1KX6-D05 (PaB670066) CDR3 PRT 86 ZZ1KX6-D05 (PaB670066) VL DNA 87 ZZ1KX6-D05 (PaB670066) VL PRT 42 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 88 ZZ1KX6-D05 (PaB670066) CDR1 PRT 89 ZZ1KX6-D05 (PaB670066) CDR2 PRT 90 ZZ1KX6-D05 (PaB670066) CDR3 PRT 91 ZZ1KXE-A05 (PaB670067) VH DNA 92 ZZ1KXE-A05 (PaB670067) VH PRT 93 ZZ1KXE-A05 (PaB670067) CDR1 PRT 94 ZZ1KXE-A05 (PaB670067) CDR2 PRT 95 ZZ1KXE-A05 (PaB670067) CDR3 PRT 96 ZZ1KXE-A05 (PaB670067) VL DNA 97 ZZ1KXE-A05 (PaB670067) VL PRT 98 ZZ1KXE-A05 (PaB670067) CDR1 PRT 99 ZZ1KXE-A05 (PaB670067) CDR2 PRT 100 ZZ1KXE-A05 (PaB670067) CDR3 PRT 101 ZZ1KXE-B01 (PaB670068) VH DNA 102 ZZ1KXE-B01 (PaB670068) VH PRT 103 ZZ1KXE-B01 (PaB670068) CDR1 PRT 104 ZZ1KXE-B01 (PaB670068) CDR2 PRT 105 ZZ1KXE-B01 (PaB670068) CDR3 PRT 106 ZZ1KXE-B01 (PaB670068) VL DNA 107 ZZ1KXE-B01 (PaB670068) VL PRT 108 ZZ1KXE-B01 (PaB670068) CDR1 PRT 109 ZZ1KXE-B01 (PaB670068) CDR2 PRT 110 ZZ1KXE-B01 (PaB670068) CDR3 PRT 111 ZZ1KXE-D06 (PaB670070) VH DNA 112 ZZ1KXE-D06 (PaB670070) VH PRT 113 ZZ1KXE-D06 (PaB670070) CDR1 PRT 114 ZZ1KXE-D06 (PaB670070) CDR2 PRT 115 ZZ1KXE-D06 (PaB670070) CDR3 PRT 116 ZZ1KXE-D06 (PaB670070) VL DNA 117 ZZ1KXE-D06 (PaB670070) VL PRT 118 ZZ1KXE-D06 (PaB670070) CDR1 PRT 119 ZZ1KXE-D06 (PaB670070) CDR2 PRT 120 ZZ1KXE-D06 (PaB670070) CDR3 PRT 121 ZZ1L3F-A02 (PaB670071) VH DNA 122 ZZ1L3F-A02 (PaB670071) VH PRT 123 ZZ1L3F-A02 (PaB670071) CDR1 PRT 124 ZZ1L3F-A02 (PaB670071) CDR2 PRT 125 ZZ1L3F-A02 (PaB670071) CDR3 PRT 126 ZZ1L3F-A02 (PaB670071) VL DNA 127 ZZ1L3F-A02 (PaB670071) VL PRT 128 ZZ1L3F-A02 (PaB670071) CDR1 PRT 129 ZZ1L3F-A02 (PaB670071) CDR2 PRT 130 ZZ1L3F-A02 (PaB670071) CDR3 PRT 131 ZZ1L3F-H03 (PaB670073) VH DNA 132 ZZ1L3F-H03 (PaB670073) VH PRT 133 ZZ1L3F-H03 (PaB670073) CDR1 PRT 134 ZZ1L3F-H03 (PaB670073) CDR2 PRT 135 ZZ1L3F-H03 (PaB670073) CDR3 PRT 136 ZZ1L3F-H03 (PaB670073) VL DNA 137 ZZ1L3F-H03 (PaB670073) VL PRT 43 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 138 ZZ1L3F-H03 (PaB670073) CDR1 PRT 139 ZZ1L3F-H03 (PaB670073) CDR2 PRT 140 ZZ1L3F-H03 (PaB670073) CDR3 PRT 141 ZZ1NHH-A05 (PaB670075) VH DNA 142 ZZ1NHH-A05 (PaB670075) VH PRT 143 ZZ1NHH-A05 (PaB670075) CDR1 PRT 144 ZZ1NHH-A05 (PaB670075) CDR2 PRT 145 ZZ1NHH-A05 (PaB670075) CDR3 PRT 146 ZZ1NHH-A05 (PaB670075) VL DNA 147 ZZ1NHH-A05 (PaB670075) VL PRT 148 ZZ1NHH-A05 (PaB670075) CDR1 PRT 149 ZZ1NHH-A05 (PaB670075) CDR2 PRT 150 ZZ1NHH-A05 (PaB670075) CDR3 PRT 151 ZZ1NHH-F09 (PaB670076) VH DNA 152 ZZ1NHH-F09 (PaB670076) VH PRT 153 ZZ1NHH-F09 (PaB670076) CDR1 PRT 154 ZZ1NHH-F09 (PaB670076) CDR2 PRT 155 ZZ1NHH-F09 (PaB670076) CDR3 PRT 156 ZZ1NHH-F09 (PaB670076) VL DNA 157 ZZ1NHH-F09 (PaB670076) VL PRT 158 ZZ1NHH-F09 (PaB670076) CDR1 PRT 159 ZZ1NHH-F09 (PaB670076) CDR2 PRT 160 ZZ1NHH-F09 (PaB670076) CDR3 PRT 161 ZZ1OZJ-C11 (PaB670077) VH DNA 162 ZZ1OZJ-C11 (PaB670077) VH PRT 163 ZZ1OZJ-C11 (PaB670077) CDR1 PRT 164 ZZ1OZJ-C11 (PaB670077) CDR2 PRT 165 ZZ1OZJ-C11 (PaB670077) CDR3 PRT 166 ZZ1OZJ-C11 (PaB670077) VL DNA 167 ZZ1OZJ-C11 (PaB670077) VL PRT 168 ZZ1OZJ-C11 (PaB670077) CDR1 PRT 169 ZZ1OZJ-C11 (PaB670077) CDR2 PRT 170 ZZ1OZJ-C11 (PaB670077) CDR3 PRT 171 ZZ1OZJ-G03 (PaB670078) VH DNA 172 ZZ1OZJ-G03 (PaB670078) VH PRT 173 ZZ1OZJ-G03 (PaB670078) CDR1 PRT 174 ZZ1OZJ-G03 (PaB670078) CDR2 PRT 175 ZZ1OZJ-G03 (PaB670078) CDR3 PRT 176 ZZ1OZJ-G03 (PaB670078) VL DNA 177 ZZ1OZJ-G03 (PaB670078) VL PRT 178 ZZ1OZJ-G03 (PaB670078) CDR1 PRT 179 ZZ1OZJ-G03 (PaB670078) CDR2 PRT 180 ZZ1OZJ-G03 (PaB670078) CDR3 PRT 181 ZZ1OZJ-G05 (PaB670079) VH DNA 182 ZZ1OZJ-G05 (PaB670079) VH PRT 183 ZZ1OZJ-G05 (PaB670079) CDR1 PRT 184 ZZ1OZJ-G05 (PaB670079) CDR2 PRT 185 ZZ1OZJ-G05 (PaB670079) CDR3 PRT 186 ZZ1OZJ-G05 (PaB670079) VL DNA 187 ZZ1OZJ-G05 (PaB670079) VL PRT 44 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 188 ZZ1OZJ-G05 (PaB670079) CDR1 PRT 189 ZZ1OZJ-G05 (PaB670079) CDR2 PRT 190 ZZ1OZJ-G05 (PaB670079) CDR3 PRT 191 PaB670080 VH DNA 192 PaB670080 VH PRT 193 PaB670080 CDR1 PRT 194 PaB670080 CDR2 PRT 195 PaB670080 CDR3 PRT 196 PaB670080 VL DNA 197 PaB670080 VL PRT 198 PaB670080 CDR1 PRT 199 PaB670080 CDR2 PRT 200 PaB670080 CDR3 PRT 201 ZZ1OZA-C01 (PaB670081) VH DNA 202 ZZ1OZA-C01 (PaB670081) VH PRT 203 ZZ1OZA-C01 (PaB670081) CDR1 PRT 204 ZZ1OZA-C01 (PaB670081) CDR2 PRT 205 ZZ1OZA-C01 (PaB670081) CDR3 PRT 206 ZZ1OZA-C01 (PaB670081) VL DNA 207 ZZ1OZA-C01 (PaB670081) VL PRT 208 ZZ1OZA-C01 (PaB670081) CDR1 PRT 209 ZZ1OZA-C01 (PaB670081) CDR2 PRT 210 ZZ1OZA-C01 (PaB670081) CDR3 PRT 211 ZZ1OZA-D02 (PaB670082) VH DNA 212 ZZ1OZA-D02 (PaB670082) VH PRT 213 ZZ1OZA-D02 (PaB670082) CDR1 PRT 214 ZZ1OZA-D02 (PaB670082) CDR2 PRT 215 ZZ1OZA-D02 (PaB670082) CDR3 PRT 216 ZZ1OZA-D02 (PaB670082) VL DNA 217 ZZ1OZA-D02 (PaB670082) VL PRT 218 ZZ1OZA-D02 (PaB670082) CDR1 PRT 219 ZZ1OZA-D02 (PaB670082) CDR2 PRT 220 ZZ1OZA-D02 (PaB670082) CDR3 PRT 221 ZZ1OZB-H05 (PaB670083) VH DNA 222 ZZ1OZB-H05 (PaB670083) VH PRT 223 ZZ1OZB-H05 (PaB670083) CDR1 PRT 224 ZZ1OZB-H05 (PaB670083) CDR2 PRT 225 ZZ1OZB-H05 (PaB670083) CDR3 PRT 226 ZZ1OZB-H05 (PaB670083) VL DNA 227 ZZ1OZB-H05 (PaB670083) VL PRT 228 ZZ1OZB-H05 (PaB670083) CDR1 PRT 229 ZZ1OZB-H05 (PaB670083) CDR2 PRT 230 ZZ1OZB-H05 (PaB670083) CDR3 PRT 231 ZZ1PXA-A05 (PaB670084) VH DNA 232 ZZ1PXA-A05 (PaB670084) VH PRT 233 ZZ1PXA-A05 (PaB670084) CDR1 PRT 234 ZZ1PXA-A05 (PaB670084) CDR2 PRT 235 ZZ1PXA-A05 (PaB670084) CDR3 PRT 236 ZZ1PXA-A05 (PaB670084) VL DNA 237 ZZ1PXA-A05 (PaB670084) VL PRT 45 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 238 ZZ1PXA-A05 (PaB670084) CDR1 PRT 239 ZZ1PXA-A05 (PaB670084) CDR2 PRT 240 ZZ1PXA-A05 (PaB670084) CDR3 PRT 241 ZZ1ODR-A02 (PaB670085) VH DNA 242 ZZ1ODP-A02 (PaB670085) VH PRT 243 ZZ1ODR-A02 (PaB670085) CDR1 PRT 244 ZZ1ODR-A02 (PaB670085) CDR2 PRT 245 ZZ1ODR-A02 (PaB670085) CDR3 PRT 246 ZZ1ODR-A02 (PaB670085) VL DNA 247 ZZ1ODR-A02 (PaB670085) VL PRT 248 ZZ1ODR-A02 (PaB670085) CDR1 PRT 249 ZZ1ODR-A02 (PaB670085) CDR2 PRT 250 ZZ1ODR-A02 (PaB670085) CDR3 PRT 251 ZZ1ODR-B05 (PaB670087) VH DNA 252 ZZ1ODR-B05 (PaB670087) VH PRT 253 ZZ1ODR-B05 (PaB670087) CDR1 PRT 254 ZZ1ODR-B05 (PaB670087) CDR2 PRT 255 ZZ1ODR-B05 (PaB670087) CDR3 PRT 256 ZZ1ODR-B05 (PaB670087) VL DNA 257 ZZ1ODR-B05 (PaB670087) VL PRT 258 ZZ1ODR-B05 (PaB670087) CDR1 PRT 259 ZZ1ODR-B05 (PaB670087) CDR2 PRT 260 ZZ1ODR-B05 (PaB670087) CDR3 PRT 261 ZZ1ODR-B11 (PaB670088) VH DNA 262 ZZ1ODR-B11 (PaB670088) VH PRT 263 ZZ1ODR-B11 (PaB670088) CDR1 PRT 264 ZZ1ODR-B11 (PaB670088) CDR2 PRT 265 ZZ1ODR-B11 (PaB670088) CDR3 PRT 266 ZZ1ODR-B11 (PaB670088) VL DNA 267 ZZ1ODR-B11 (PaB670088) VL PRT 268 ZZ1ODR-B11 (PaB670088) CDR1 PRT 269 ZZ1ODR-B11 (PaB670088) CDR2 PRT 270 ZZ1ODR-B11 (PaB670088) CDR3 PRT 271 ZZ1ODR-C05 (PaB670089) VH DNA 272 ZZ1ODR-C05 (PaB670089) VH PRT 273 ZZ1ODR-C05 (PaB670089) CDR1 PRT 274 ZZ1ODR-C05 (PaB670089) CDR2 PRT 275 ZZ1ODR-C05 (PaB670089) CDR3 PRT 276 ZZ1ODR-C05 (PaB670089) VL DNA 277 ZZ1ODR-C05 (PaB670089) VL PRT 278 ZZ1ODR-C05 (PaB670089) CDR1 PRT 279 ZZ1ODR-C05 (PaB670089) CDR2 PRT 280 ZZ1ODR-C05 (PaB670089) CDR3 PRT 281 ZZ1ODR-F02 (PaB670090) VH DNA 282 ZZ1ODR-F02 (PaB670090) VH PRT 283 ZZ1ODP-F02 (PaB670090) CDR1 PRT 284 ZZ1ODP-F02 (PaB670090) CDR2 PRT 285 ZZ1ODR-F02 (PaB670090) CDR3 PRT 286 ZZ1ODP-F02 (PaB670090) VL DNA 287 ZZ1ODP-F02 (PaB670090) VL PRT 46 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 288 ZZ1ODR-F02 (PaB670090) CDR1 PRT 289 ZZ1ODP-F02 (PaB670090) CDR2 PRT 290 ZZ1ODP-F02 (PaB670090) CDR3 PRT 291 ZZ1ODP-G02 (PaB670091) VH DNA 292 ZZ1ODP-G02 (PaB670091) VH PRT 293 ZZ1ODP-G02 (PaB670091) CDR1 PRT 294 ZZ1ODP-G02 (PaB670091) CDR2 PRT 295 ZZ1ODP-G02 (PaB670091) CDR3 PRT 296 ZZ1ODP-G02 (PaB670091) VL DNA 297 ZZ1ODP-G02 (PaB670091) VL PRT 298 ZZ1ODP-G02 (PaB670091) CDR1 PRT 299 ZZ1ODP-G02 (PaB670091) CDR2 PRT 300 ZZ1ODP-G02 (PaB670091) CDR3 PRT 301 ZZ1ODP-G11 (PaB670092) VH DNA 302 ZZ1ODP-G11 (PaB670092) VH PRT 303 ZZ1ODP-G11 (PaB670092) CDR1 PRT 304 ZZ1ODP-G11 (PaB670092) CDR2 PRT 305 ZZ1ODP-G11 (PaB670092) CDR3 PRT 306 ZZ1ODP-G11 (PaB670092) VL DNA 307 ZZ1ODP-G11 (PaB670092) VL PRT 308 ZZ1ODP-G11 (PaB670092) CDR1 PRT 309 ZZ1ODP-G11 (PaB670092) CDR2 PRT 310 ZZ1ODP-G11 (PaB670092) CDR3 PRT 311 ZZ1ODP-H04 (PaB670093) VH DNA 312 ZZ1ODP-H04 (PaB670093) VH PRT 313 ZZ1ODP-H04 (PaB670093) CDR1 PRT 314 ZZ1ODP-H04 (PaB670093) CDR2 PRT 315 ZZ1ODP-H04 (PaB670093) CDR3 PRT 316 ZZ1ODP-H04 (PaB670093) VL DNA 317 ZZ1ODP-H04 (PaB670093) VL PRT 318 ZZ1ODP-H04 (PaB670093) CDR1 PRT 319 ZZ1ODP-H04 (PaB670093) CDR2 PRT 320 ZZ1ODP-H04 (PaB670093) CDR3 PRT 321 ZZ1ODS-B08 (PaB670094) VH DNA 322 ZZ1ODS-B08 (PaB670094) VH PRT 323 ZZ1ODS-B08 (PaB670094) CDR1 PRT 324 ZZ1ODS-B08 (PaB670094) CDR2 PRT 325 ZZ1ODS-B08 (PaB670094) CDR3 PRT 326 ZZ1ODS-B08 (PaB670094) VL DNA 327 ZZ1ODS-B08 (PaB670094) VL PRT 328 ZZ1ODS-B08 (PaB670094) CDR1 PRT 329 ZZ1ODS-B08 (PaB670094) CDR2 PRT 330 ZZ1ODS-B08 (PaB670094) CDR3 PRT 331 ZZ1ODS-H05 (PaB67009S) VH DNA 332 ZZ1ODS-H05 (PaB670095) VH PRT 333 ZZ1ODS-H05 (PaB67009S) CDR1 PRT 334 ZZ1ODS-H05 (PaB67009S) CDR2 PRT 335 ZZ1ODS-H05 (PaB67009S) CDR3 PRT 336 ZZ1ODS-H05 (PaB67009S) VL DNA 337 ZZ1ODS-H05 (PaB67009S) VL PRT 47 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 338 ZZ1ODS-H05 (PaB67009S) CDR1 PRT 339 ZZ1ODS-H05 (PaB67009S) CDR2 PRT 340 ZZ1ODS-H05 (PaB67009S) CDR3 PRT 341 ZZ1ODT-E11 (PaB670097) VH DNA 342 ZZ1ODT-E11 (PaB670097) VH PRT 343 ZZ1ODT-E11 (PaB670097) CDR1 PRT 344 ZZ1ODT-E11 (PaB670097) CDR2 PRT 345 ZZ1ODT-E11 (PaB670097) CDR3 PRT 346 ZZ1ODT-E11 (PaB670097) VL DNA 347 ZZ1ODT-E11 (PaB670097) VL PRT 348 ZZ1ODT-E11 (PaB670097) CDR1 PRT 349 ZZ1ODT-E11 (PaB670097) CDR2 PRT 350 ZZ1ODT-E11 (PaB670097) CDR3 PRT 351 ZZ1ODT-G01 (PaB670098) VH DNA 352 ZZ1ODT-G01 (PaB670098) VH PRT 353 ZZ1ODT-G01 (PaB670098) CDR1 PRT 354 ZZ1ODT-G01 (PaB670098) CDR2 PRT 355 ZZ1ODT-G01 (PaB670098) CDR3 PRT 356 ZZ1ODT-G01 (PaB670098) VL DNA 357 ZZ1ODT-G01 (PaB670098) VL PRT 358 ZZ1ODT-G01 (PaB670098) CDR1 PRT 359 ZZ1ODT-G01 (PaB670098) CDR2 PRT 360 ZZ1ODT-G01 (PaB670098) CDR3 PRT 361 PaB670099 VH DNA 362 PaB670099 VH PRT 363 PaB670099 CDR1 PRT 364 PaB670099 CDR2 PRT 365 PaB670099 CDR3 PRT 366 PaB670099 VL DNA 367 PaB670099 VL PRT 368 PaB670099 CDR1 PRT 369 PaB670099 CDR2 PRT 370 PaB670099 CDR3 PRT 371 PaB670100 VH DNA 372 PaB670100 VH PRT 373 PaB670100 CDR1 PRT 374 PaB670100 CDR2 PRT 375 PaB670100 CDR3 PRT 376 PaB670100 VL DNA 377 PaB670100 VL PRT 378 PaB670100 CDR1 PRT 379 PaB670100 CDR2 PRT 380 PaB670100 CDR3 PRT 381 ZZ1ODO-H01 (PaB670101) VH DNA 382 ZZ1ODO-H01 (PaB670101) VH PRT 383 ZZ1ODO-H01 (PaB670101) CDR1 PRT 384 ZZ1ODO-H01 (PaB670101) CDR2 PRT 385 ZZ1ODO-H01 (PaB670101) CDR3 PRT 386 ZZ1ODO-H01 (PaB670101) VL DNA 387 ZZ1ODO-H01 (PaB670101) VL PRT 48 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 388 ZZ1ODO-H01 (PaB670101) CDR1 PRT 389 ZZ1ODO-H01 (PaB670101) CDR2 PRT 390 ZZ1ODO-H01 (PaB670101) CDR3 PRT 391 ZZ1PXS-F08 (PaB670102) VH DNA 392 ZZ1PXS-F08 (PaB670102) VH PRT 393 ZZ1PXS-F08 (PaB670102) CDR1 PRT 394 ZZ1PXS-F08 (PaB670102) CDR2 PRT 395 ZZ1PXS-F08 (PaB670102) CDR3 PRT 396 ZZ1PXS-F08 (PaB670102) VL DNA 397 ZZ1PXS-F08 (PaB670102) VL PRT 398 ZZ1PXS-F08 (PaB670102) CDR1 PRT 399 ZZ1PXS-F08 (PaB670102) CDR2 PRT 400 ZZ1PXS-F08 (PaB670102) CDR3 PRT 401 ZZ1RCX-C09 (PaB670103) VH DNA 402 ZZ1RCX-C09 (PaB670103) VH PRT 403 ZZ1RCX-C09 (PaB670103) CDR1 PRT 404 ZZ1RCX-C09 (PaB670103) CDR2 PRT 405 ZZ1RCX-C09 (PaB670103) CDR3 PRT 406 ZZ1RCX-C09 (PaB670103) VL DNA 407 ZZ1RCX-C09 (PaB670103) VL PRT 408 ZZ1RCX-C09 (PaB670103) CDR1 PRT 409 ZZ1RCX-C09 (PaB670103) CDR2 PRT 410 ZZ1RCX-C09 (PaB670103) CDR3 PRT 411 PaB670104 VH DNA 412 PaB670104 VH PRT 413 PaB670104 CDR1 PRT 414 PaB670104 CDR2 PRT 415 PaB670104 CDR3 PRT 416 PaB670104 VL DNA 417 PaB670104 VL PRT 418 PaB670104 CDR1 PRT 419 PaB670104 CDR2 PRT 420 PaB670104 CDR3 PRT 421 ZZ1RD0-D01 (PaB670105) VH DNA 422 ZZ1RD0-D01 (PaB670105) VH PRT 423 ZZ1RD0-D01 (PaB670105) CDR1 PRT 424 ZZ1RD0-D01 (PaB670105) CDR2 PRT 425 ZZ1RD0-D01 (PaB670105) CDR3 PRT 426 ZZ1RD0-D01 (PaB670105) VL DNA 427 ZZ1RD0-D01 (PaB670105) VL PRT 428 ZZ1RD0-D01 (PaB670105) CDR1 PRT 429 ZZ1RD0-D01 (PaB670105) CDR2 PRT 430 ZZ1RD0-D01 (PaB670105) CDR3 PRT 431 ZZ1RD0-G02 (PaB670106) VH DNA 432 ZZ1RD0-G02 (PaB670106) VH PRT 433 ZZ1RD0-G02 (PaB670106) CDR1 PRT 434 ZZ1RD0-G02 (PaB670106) CDR2 PRT 435 ZZ1RD0-G02 (PaB670106) CDR3 PRT 436 ZZ1RD0-G02 (PaB670106) VL DNA 437 ZZ1RD0-G02 (PaB670106) VL PRT 49 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 438 ZZ1RD0-G02 (PaB670106) CDR1 PRT 439 ZZ1RD0-G02 (PaB670106) CDR2 PRT 440 ZZ1RD0-G02 (PaB670106) CDR3 PRT 441 ZZ1RD3-D09 (PaB670107) VH DNA 442 ZZ1RD3-D09 (PaB670107) VH PRT 443 ZZ1RD3-D09 (PaB670107) CDR1 PRT 444 ZZ1RD3-D09 (PaB670107) CDR2 PRT 445 ZZ1RD3-D09 (PaB670107) CDR3 PRT 446 ZZ1RD3-D09 (PaB670107) VL DNA 447 ZZ1RD3-D09 (PaB670107) VL PRT 448 ZZ1RD3-D09 (PaB670107) CDR1 PRT 449 ZZ1RD3-D09 (PaB670107) CDR2 PRT 450 ZZ1RD3-D09 (PaB670107) CDR3 PRT 451 ZZ1RD3-H03 (PaB670108) VH DNA 452 ZZ1RD3-H03 (PaB670108) VH PRT 453 ZZ1RD3-H03 (PaB670108) CDR1 PRT 454 ZZ1RD3-H03 (PaB670108) CDR2 PRT 455 ZZ1RD3-H03 (PaB670108) CDR3 PRT 456 ZZ1RD3-H03 (PaB670108) VL DNA 457 ZZ1RD3-H03 (PaB670108) VL PRT 458 ZZ1RD3-H03 (PaB670108) CDR1 PRT 459 ZZ1RD3-H03 (PaB670108) CDR2 PRT 460 ZZ1RD3-H03 (PaB670108) CDR3 PRT 461 ZZ1RUC-C01 (PaB670114) VH DNA 462 ZZ1RUC-C01 (PaB670114) VH PRT 463 ZZ1RUC-C01 (PaB670114) CDR1 PRT 464 ZZ1RUC-C01 (PaB670114) CDR2 PRT 465 ZZ1RUC-C01 (PaB670114) CDR3 PRT 466 ZZ1RUC-C01 (PaB670114) VL DNA 467 ZZ1RUC-C01 (PaB670114) VL PRT 468 ZZ1RUC-C01 (PaB670114) CDR1 PRT 469 ZZ1RUC-C01 (PaB670114) CDR2 PRT 470 ZZ1RUC-C01 (PaB670114) CDR3 PRT 471 ZZ1RUC-G02 (PaB670115) VH DNA 472 ZZ1RUC-G02 (PaB670115) VH PRT 473 ZZ1RUC-G02 (PaB670115) CDR1 PRT 474 ZZ1RUC-G02 (PaB670115) CDR2 PRT 475 ZZ1RUC-G02 (PaB670115) CDR3 PRT 476 ZZ1RUC-G02 (PaB670115) VL DNA 477 ZZ1RUC-G02 (PaB670115) VL PRT 478 ZZ1RUC-G02 (PaB670115) CDR1 PRT 479 ZZ1RUC-G02 (PaB670115) CDR2 PRT 480 ZZ1RUC-G02 (PaB670115) CDR3 PRT 481 ZZ1RUC-B04 (PaB670116) VH DNA 482 ZZ1RUC-B04 (PaB670116) VH PRT 483 ZZ1RUC-B04 (PaB670116) CDR1 PRT 484 ZZ1RUC-B04 (PaB670116) CDR2 PRT 485 ZZ1RUC-B04 (PaB670116) CDR3 PRT 486 ZZ1RUC-B04 (PaB670116) VL DNA 487 ZZ1RUC-B04 (PaB670116) VL PRT 50 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 488 ZZ1RUC-B04 (PaB670116) CDR1 PRT 489 ZZ1RUC-B04 (PaB670116) CDR2 PRT 490 ZZ1RUC-B04 (PaB670116) CDR3 PRT 491 ZZ1RUC-A06 (PaB670117) VH DNA 492 ZZ1RUC-A06 (PaB670117) VH PRT 493 ZZ1RUC-A06 (PaB670117) CDR1 PRT 494 ZZ1RUC-A06 (PaB670117) CDR2 PRT 495 ZZ1RUC-A06 (PaB670117) CDR3 PRT 496 ZZ1RUC-A06 (PaB670117) VL DNA 497 ZZ1RUC-A06 (PaB670117) VL PRT 498 ZZ1RUC-A06 (PaB670117) CDR1 PRT 499 ZZ1RUC-A06 (PaB670117) CDR2 PRT 500 ZZ1RUC-A06 (PaB670117) CDR3 PRT 501 ZZ1RUC-A07 (PaB670118) VH DNA 502 ZZ1RUC-A07 (PaB670118) VH PRT 503 ZZ1RUC-A07 (PaB670118) CDR1 PRT 504 ZZ1RUC-A07 (PaB670118) CDR2 PRT 505 ZZ1RUC-A07 (PaB670118) CDR3 PRT 506 ZZ1RUC-A07 (PaB670118) VL DNA 507 ZZ1RUC-A07 (PaB670118) VL PRT 508 ZZ1RUC-A07 (PaB670118) CDR1 PRT 509 ZZ1RUC-A07 (PaB670118) CDR2 PRT 510 ZZ1RUC-A07 (PaB670118) CDR3 PRT 511 ZZ1RUC-G08 (PaB670119) VH DNA 512 ZZ1RUC-G08 (PaB670119) VH PRT 513 ZZ1RUC-G08 (PaB670119) CDR1 PRT 514 ZZ1RUC-G08 (PaB670119) CDR2 PRT 515 ZZ1RUC-G08 (PaB670119) CDR3 PRT 516 ZZ1RUC-G08 (PaB670119) VL DNA 517 ZZ1RUC-G08 (PaB670119) VL PRT 518 ZZ1RUC-G08 (PaB670119) CDR1 PRT 519 ZZ1RUC-G08 (PaB670119) CDR2 PRT 520 ZZ1RUC-G08 (PaB670119) CDR3 PRT 521 ZZ1RUC-C11 (PaB670120) VH DNA 522 ZZ1RUC-C11 (PaB670120) VH PRT 523 ZZ1RUC-C11 (PaB670120) CDR1 PRT 524 ZZ1RUC-C11 (PaB670120) CDR2 PRT 525 ZZ1RUC-C11 (PaB670120) CDR3 PRT 526 ZZ1RUC-C11 (PaB670120) VL DNA 527 ZZ1RUC-C11 (PaB670120) VL PRT 528 ZZ1RUC-C11 (PaB670120) CDR1 PRT 529 ZZ1RUC-C11 (PaB670120) CDR2 PRT 530 ZZ1RUC-C11 (PaB670120) CDR3 PRT 531 ZZ1RUC-H11 (PaB670121) VH DNA 532 ZZ1RUC-H11 (PaB670121) VH PRT 533 ZZ1RUC-H11 (PaB670121) CDR1 PRT 534 ZZ1RUC-H11 (PaB670121) CDR2 PRT 535 ZZ1RUC-H11 (PaB670121) CDR3 PRT 536 ZZ1RUC-H11 (PaB670121) VL DNA 537 ZZ1RUC-H11 (PaB670121) VL PRT 51 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 538 ZZ1RUC-H11 (PaB670121) CDR1 PRT 539 ZZ1RUC-H11 (PaB670121) CDR2 PRT 540 ZZ1RUC-H11 (PaB670121) CDR3 PRT 541 ZZ1RUD-A02 (PaB670122) VH DNA 542 ZZ1RUD-A02 (PaB670122) VH PRT 543 ZZ1RUD-A02 (PaB670122) CDR1 PRT 544 ZZ1RUD-A02 (PaB670122) CDR2 PRT 545 ZZ1RUD-A02 (PaB670122) CDR3 PRT 546 ZZ1RUD-A02 (PaB670122) VL DNA 547 ZZ1RUD-A02 (PaB670122) VL PRT 548 ZZ1RUD-A02 (PaB670122) CDR1 PRT 549 ZZ1RUD-A02 (PaB670122) CDR2 PRT 550 ZZ1RUD-A02 (PaB670122) CDR3 PRT 551 ZZ1RUD-H03 (PaB670123) VH DNA 552 ZZ1RUD-H03 (PaB670123) VH PRT 553 ZZ1RUD-H03 (PaB670123) CDR1 PRT 554 ZZ1RUD-H03 (PaB670123) CDR2 PRT 555 ZZ1RUD-H03 (PaB670123) CDR3 PRT 556 ZZ1RUD-H03 (PaB670123) VL DNA 557 ZZ1RUD-H03 (PaB670123) VL PRT 558 ZZ1RUD-H03 (PaB670123) CDR1 PRT 559 ZZ1RUD-H03 (PaB670123) CDR2 PRT 560 ZZ1RUD-H03 (PaB670123) CDR3 PRT 561 ZZ1RUD-H06 (PaB670125) VH DNA 562 ZZ1RUD-H06 (PaB670125) VH PRT 563 ZZ1RUD-H06 (PaB670125) CDR1 PRT 564 ZZ1RUD-H06 (PaB670125) CDR2 PRT 565 ZZ1RUD-H06 (PaB670125) CDR3 PRT 566 ZZ1RUD-H06 (PaB670125) VL DNA 567 ZZ1RUD-H06 (PaB670125) VL PRT 568 ZZ1RUD-H06 (PaB670125) CDR1 PRT 569 ZZ1RUD-H06 (PaB670125) CDR2 PRT 570 ZZ1RUD-H06 (PaB670125) CDR3 PRT 571 ZZ1RUD-B08 (PaB670126) VH DNA 572 ZZ1RUD-B08 (PaB670126) VH PRT 573 ZZ1RUD-B08 (PaB670126) CDR1 PRT 574 ZZ1RUD-B08 (PaB670126) CDR2 PRT 575 ZZ1RUD-B08 (PaB670126) CDR3 PRT 576 ZZ1RUD-B08 (PaB670126) VL DNA 577 ZZ1RUD-B08 (PaB670126) VL PRT 578 ZZ1RUD-B08 (PaB670126) CDR1 PRT 579 ZZ1RUD-B08 (PaB670126) CDR2 PRT 580 ZZ1RUD-B08 (PaB670126) CDR3 PRT 581 ZZ1RUD-H10 (PaB670127) VH DNA 582 ZZ1RUD-H10 (PaB670127) VH PRT 583 ZZ1RUD-H10 (PaB670127) CDR1 PRT 584 ZZ1RUD-H10 (PaB670127) CDR2 PRT 585 ZZ1RUD-H10 (PaB670127) CDR3 PRT 586 ZZ1RUD-H10 (PaB670127) VL DNA 587 ZZ1RUD-H10 (PaB670127) VL PRT 52 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 588 ZZ1RUD-H10 (PaB670127) CDR1 PRT 589 ZZ1RUD-H10 (PaB670127) CDR2 PRT 590 ZZ1RUD-H10 (PaB670127) CDR3 PRT 591 ZZ1RUE-A01 (PaB670128) VH DNA 592 ZZ1RUE-A01 (PaB670128) VH PRT 593 ZZ1RUE-A01 (PaB670128) CDR1 PRT 594 ZZ1RUE-A01 (PaB670128) CDR2 PRT 595 ZZ1RUE-A01 (PaB670128) CDR3 PRT 596 ZZ1RUE-A01 (PaB670128) VL DNA 597 ZZ1RUE-A01 (PaB670128) VL PRT 598 ZZ1RUE-A01 (PaB670128) CDR1 PRT 599 ZZ1RUE-A01 (PaB670128) CDR2 PRT 600 ZZ1RUE-A01 (PaB670128) CDR3 PRT 601 ZZ1RUF-A01 (PaB670136) VH DNA 602 ZZ1RUF-A01 (PaB670136) VH PRT 603 ZZ1RUF-A01 (PaB670136) CDR1 PRT 604 ZZ1RUF-A01 (PaB670136) CDR2 PRT 605 ZZ1RUF-A01 (PaB670136) CDR3 PRT 606 ZZ1RUF-A01 (PaB670136) VL DNA 607 ZZ1RUF-A01 (PaB670136) VL PRT 608 ZZ1RUF-A01 (PaB670136) CDR1 PRT 609 ZZ1RUF-A01 (PaB670136) CDR2 PRT 610 ZZ1RUF-A01 (PaB670136) CDR3 PRT 611 ZZ1RUF-B06 (PaB670137) VH DNA 612 ZZ1RUF-B06 (PaB670137) VH PRT 613 ZZ1RUF-B06 (PaB670137) CDR1 PRT 614 ZZ1RUF-B06 (PaB670137) CDR2 PRT 615 ZZ1RUF-B06 (PaB670137) CDR3 PRT 616 ZZ1RUF-B06 (PaB670137) VL DNA 617 ZZ1RUF-B06 (PaB670137) VL PRT 618 ZZ1RUF-B06 (PaB670137) CDR1 PRT 619 ZZ1RUF-B06 (PaB670137) CDR2 PRT 620 ZZ1RUF-B06 (PaB670137) CDR3 PRT 621 PaB670141 VH DNA 622 PaB670141 VH PRT 623 PaB670141 CDR1 PRT 624 PaB670141 CDR2 PRT 625 PaB670141 CDR3 PRT 626 PaB670141 VL DNA 627 PaB670141 VL PRT 628 PaB670141 CDR1 PRT 629 PaB670141 CDR2 PRT 630 PaB670141 CDR3 PRT 631 PaB670142 VH DNA 632 PaB670142 VH PRT 633 PaB670142 CDR1 PRT 634 PaB670142 CDR2 PRT 635 PaB670142 CDR3 PRT 636 PaB670142 VL DNA 637 PaB670142 VL PRT 53 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 638 PaB670142 CDR1 PRT 639 PaB670142 CDR2 PRT 640 PaB670142 CDR3 PRT 641 PaB670143 VH DNA 642 PaB670143 VH PRT 643 PaB670143 CDR1 PRT 644 PaB670143 CDR2 PRT 645 PaB670143 CDR3 PRT 646 PaB670143 VL DNA 647 PaB670143 VL PRT 648 PaB670143 CDR1 PRT 649 PaB670143 CDR2 PRT 650 PaB670143 CDR3 PRT 651 PaB670144 VH DNA 652 PaB670144 VH PRT 653 PaB670144 CDR1 PRT 654 PaB670144 CDR2 PRT 655 PaB670144 CDR3 PRT 656 PaB670144 VL DNA 657 PaB670144 VL PRT 658 PaB670144 CDR1 PRT 659 PaB670144 CDR2 PRT 660 PaB670144 CDR3 PRT 661 PaB670146 VH DNA 662 PaB670146 VH PRT 663 PaB670146 CDR1 PRT 664 PaB670146 CDR2 PRT 665 PaB670146 CDR3 PRT 666 PaB670146 VL DNA 667 PaB670146 VL PRT 668 PaB670146 CDR1 PRT 669 PaB670146 CDR2 PRT 670 PaB670146 CDR3 PRT 671 PaB670148 VH DNA 672 PaB670148 VH PRT 673 PaB670148 CDR1 PRT 674 PaB670148 CDR2 PRT 675 PaB670148 CDR3 PRT 676 PaB670148 VL DNA 677 PaB670148 VL PRT 678 PaB670148 CDR1 PRT 679 PaB670148 CDR2 PRT 680 PaB670148 CDR3 PRT 681 PaB670149 VH DNA 682 PaB670149 VH PRT 683 PaB670149 CDR1 PRT 684 PaB670149 CDR2 PRT 685 PaB670149 CDR3 PRT 686 PaB670149 VL DNA 687 PaB670149 VL PRT 54 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 688 PaB670149 CDR1 PRT 689 PaB670149 CDR2 PRT 690 PaB670149 CDR3 PRT 691 PaB670151 VH DNA 692 PaB670151 VH PRT 693 PaB670151 CDR1 PRT 694 PaB670151 CDR2 PRT 695 PaB670151 CDR3 PRT 696 PaB670151 VL DNA 697 PaB670151 VL PRT 698 PaB670151 CDR1 PRT 699 PaB670151 CDR2 PRT 700 PaB670151 CDR3 PRT 701 PaB670152 VH DNA 702 PaB670152 VH PRT 703 PaB670152 CDR1 PRT 704 PaB670152 CDR2 PRT 705 PaB670152 CDR3 PRT 706 PaB670152 VL DNA 707 PaB670152 VL PRT 708 PaB670152 CDR1 PRT 709 PaB670152 CDR2 PRT 710 PaB670152 CDR3 PRT 711 PaB670153 VH DNA 712 PaB670153 VH PRT 713 PaB670153 CDR1 PRT 714 PaB670153 CDR2 PRT 715 PaB670153 CDR3 PRT 716 PaB670153 VL DNA 717 PaB670153 VL PRT 718 PaB670153 CDR1 PRT 719 PaB670153 CDR2 PRT 720 PaB670153 CDR3 PRT 721 PaB670156 VH DNA 722 PaB670156 VH PRT 723 PaB670156 CDR1 PRT 724 PaB670156 CDR2 PRT 725 PaB670156 CDR3 PRT 726 PaB670156 VL DNA 727 PaB670156 VL PRT 728 PaB670156 CDR1 PRT 729 PaB670156 CDR2 PRT 730 PaB670156 CDR3 PRT 731 PaB670157 VH DNA 732 PaB670157 VH PRT 733 PaB670157 CDR1 PRT 734 PaB670157 CDR2 PRT 735 PaB670157 CDR3 PRT 736 PaB670157 VL DNA 737 PaB670157 VL PRT 55 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 738 PaB670157 CDR1 PRT 739 PaB670157 CDR2 PRT 740 PaB670157 CDR3 PRT 741 PaB670158 VH DNA 742 PaB670158 VH PRT 743 PaB670158 CDR1 PRT 744 PaB670158 CDR2 PRT 745 PaB670158 CDR3 PRT 746 PaB670158 VL DNA 747 PaB670158 VL PRT 748 PaB670158 CDR1 PRT 749 PaB670158 CDR2 PRT 750 PaB670158 CDR3 PRT 751 PaB670159 VH DNA 752 PaB670159 VH PRT 753 PaB670159 CDR1 PRT 754 PaB670159 CDR2 PRT 755 PaB670159 CDR3 PRT 756 PaB670159 VL DNA 757 PaB670159 VL PRT 758 PaB670159 CDR1 PRT 759 PaB670159 CDR2 PRT 760 PaB670159 CDR3 PRT 761 PaB670160 VH DNA 762 PaB670160 VH PRT 763 PaB670160 CDR1 PRT 764 PaB670160 CDR2 PRT 765 PaB670160 CDR3 PRT 766 PaB670160 VL DNA 767 PaB670160 VL PRT 768 PaB670160 CDR1 PRT 769 PaB670160 CDR2 PRT 770 PaB670160 CDR3 PRT 771 PaB670161 VH DNA 772 PaB670161 VH PRT 773 PaB670161 CDR1 PRT 774 PaB670161 CDR2 PRT 775 PaB670161 CDR3 PRT 776 PaB670161 VL DNA 777 PaB670161 VL PRT 778 PaB670161 CDR1 PRT 779 PaB670161 CDR2 PRT 780 PaB670161 CDR3 PRT 781 PaB670162 VH DNA 782 PaB670162 VH PRT 783 PaB670162 CDR1 PRT 784 PaB670162 CDR2 PRT 785 PaB670162 CDR3 PRT 786 PaB670162 VL DNA 787 PaB670162 VL PRT 56 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (continued) SEQ ID NO: clone name Type 788 PaB670162 CDR1 PRT 789 PaB670162 CDR2 PRT 790 PaB670162 CDR3 PRT 791 PaB670163 VH DNA 792 PaB670163 VH PRT 793 PaB670163 CDR1 PRT 794 PaB670163 CDR2 PRT 795 PaB670163 CDR3 PRT 796 PaB670163 VL DNA 797 PaB670163 VL PRT 798 PaB670163 CDR1 PRT 799 PaB670163 CDR2 PRT 800 PaB670163 CDR3 PRT SEQ ID NO: 801‑ Human PAR2 Preproprotein (GenBank Accession No. NP_005233.3) SEQ ID NO: 802‑ Human PAR2 Tethered Ligand SLIGKVDGTSHVTGKGVTVETVFSVDEFSASVLTGKLTT SEQ ID NO: 803‑ Exemplary VH Framework Region 1 EVQLLESGGGLVQPGGSLRLSCAASGFTFS SEQ ID NO: 804‑ Exemplary VH Framework Region 2 WVRQAPGKGLEWVS SEQ ID NO: 805‑ Exemplary VH Framework Region 3 RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR SEQ ID NO: 806‑ Exemplary VH Framework Region 4 WGQGTLVTVSS SEQ ID NO: 807‑ Exemplary VL Framework Region 1 SIELTQPPSVSVSPGQTASITC SEQ ID NO: 808‑ Exemplary VL Framework Region 2 WYQQKPGQSPVLVIY SEQ ID NO: 809‑ Exemplary VL Framework Region 3 GIPERFSGSNSGNTATLTISGTQAMDEADYYC SEQ ID NO: 810‑ Exemplary VL Framework Region 4 FGGGTKLTVL SEQ ID NO: 811‑ Exemplary VH CDR2 TISYSGSHISYHDSVHH SEQ ID NO: 812‑ Exemplary VH CDR2 TISYHGSLISYHDSVHH SEQ ID NO: 813‑ Exemplary VH CDR2 TISYHGSHISYADSVHH SEQ ID NO: 814‑ Exemplary VH CDR2 TISYHGSHISYHDSVKH SEQ ID NO: 815‑ Exemplary VH CDR2 57 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 TISYHGSHISYHDSVHG SEQ ID NO: 816- Exemplary VH CDR2 TISYHGSLISYADSVKG SEQ ID NO: 817‑ Exemplary VH CDR2 TISYSGSHISYADSVKG SEQ ID NO: 818‑ Exemplary VH CDR2 TISYHGSHISYADSVKG SEQ ID NO: 819‑ Exemplary VH CDR3 IHNDPMDV SEQ ID NO: 820‑ Exemplary VH CDR3 INHDPMDV SEQ ID NO: 821-‑ Exemplary VH SEQ ID NO: 822 -‑ Exemplary VH SEQ ID NO: 823 -‑ Exemplary VH SEQ ID NO: 824 -‑ Exemplary VH SEQ ID NO: 825 -- Exemplary VH SEQ ID NO: 826 -‑ Exemplary VH SEQ ID NO: 827 -‑ Exemplary VH SEQ ID NO: 828 -‑ Exemplary VH 58 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 SEQ ID NO: 829 -‑ Exemplary VH SEQ ID NO: 830 -‑ Exemplary VH SEQ ID NO: 831 -‑ Exemplary VH INCORPORATION BY REFERENCE

[0197] All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference.

[0198] While specific embodiments of the subject disclosure havebeen discussed, the above specification is illustrative and not restrictive. Many variations of the disclosure will become apparent to those skilled in the art upon review of this specification and the claims below. The full scope of the disclosure should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations. ASPECTS OF THE INVENTION

[0199] 1. An antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 3; ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 4; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 5; and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 8; ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 9; and iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 10; wherein the amino acid substitutions, deletions or insertions reduce the binding affinity of the antibody or antigen- binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as 59 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 compared to an antibody or antigen-binding fragment having a VHwith an amino acid sequence of SEQ IDNO: 2 and VL with an amino acid sequence of SEQ ID NO: 7 when tested at a pH of 7.4 in a PAR2 binding assay. 2. The antibody or antigen-binding fragment thereof of aspect 1, wherein the amino acid substitutions, deletions or insertions comprises a homologous substitution. 3. An antibody or antigen-binding fragment thereof comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3; ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑17 of SEQ ID NO: 4; and iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑8 of SEQ ID NO: 5; and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8; ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9; and iii) a VL-CDR3 having the amino acid sequence of SEQ IDNO: 10; but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑14 of SEQ ID NO: 10. 4. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8, ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9, iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10. 5. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH comprises: i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 13, ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 14, iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 15, and wherein the VL comprises: i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 18, ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 19, iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 20. 6. The antibody or antigen-binding fragment thereof of aspect 3, wherein a histidine is present at the amino acid positions corresponding to positions 5, 8, 12, 16, and 17 of SEQ ID NO: 4; and wherein a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ ID NO: 5. 7. Theantibody or antigen-binding fragment thereof of anyoneof aspects 3 or 5‑6,wherein a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 4. 8. Theantibody or antigen-binding fragment thereof of anyoneof aspects 3 or 5‑7,wherein a histidine is present at the amino acid position corresponding to position 7 of SEQ ID NO: 4. 9. Theantibody or antigen-binding fragment thereof of anyoneof aspects 3 or 5‑7,wherein a histidine is present at the amino acid position corresponding to position 8 of SEQ ID NO: 4. 60 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 10. The antibody or antigen-binding fragment thereof of any one of aspects 1 or 5‑7, wherein a histidine is present at the amino acid position corresponding to position 12 of SEQ ID NO: 4. 11. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑10, wherein a histidine is present at the amino acid position corresponding to position 15 of SEQ ID NO: 4. 12. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑11, wherein a histidine is present at the amino acid position corresponding to position 16 of SEQ ID NO: 4. 13. The antibody or antigen-binding fragment thereof of any one of aspects 1 or 5‑11, wherein a histidine is present at the amino acid position corresponding to position 17 of SEQ ID NO: 4. 14. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑11, wherein a histidine is present at the amino acid positions corresponding to positions 5 and 8 of SEQ ID NO: 4. 15. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑11, wherein a histidine is present at the amino acid positions corresponding to positions 5, 8, 12, and 16 of SEQ ID NO: 4. 16. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑11, wherein a histidine is present at the amino acid positions corresponding to positions 5, 8, 12, 16 and 17 of SEQ ID NO: 4. 17. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑16, wherein a histidine is present at the amino acid position corresponding to position 2 of SEQ ID NO: 5. 18. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑17, wherein a histidine is present at the amino acid position corresponding to position 3 of SEQ ID NO: 5. 19. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑18, wherein a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ ID NO: 5. 20. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑19, wherein a histidine is present at the amino acid position corresponding to position 1 of SEQ ID NO: 10. 21. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑20, wherein a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 10. 22. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑21, wherein a histidine is present at the amino acid position corresponding to position 6 of SEQ ID NO: 10. 23. The antibody or antigen-binding fragment thereof of any one of aspects 3 or 5‑22, wherein a histidine is present at the amino acid position corresponding to position 14 of SEQ ID NO: 10. 24. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH-CDR2 comprises an amino acid sequence selected from the group consisting of: SEQ IDNOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794, and 811‑818. 25. The antibody or antigen-binding fragment thereof of aspect 3 or 24, wherein the VH-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ IDNOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, and 819‑820. 26. The antibody or antigen-binding fragment thereof of any one of aspects 3, 24 or 25, wherein the VL-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ IDNOs: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 61 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790 and 800. 27. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 14; wherein the VH-CDR3 comprises an amino acid sequence correspond- ing to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 20. 28. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH-CDR2 comprises an amino acid sequence corresponding to SEQ IDNO: 811; wherein theVH-CDR3 comprises an amino acid sequence correspond- ing to SEQ IDNO: 819, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ IDNO: 10 or 20. 29. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 814; wherein the VH-CDR3 comprises an amino acid sequence corre- sponding toSEQ IDNO:820, andwherein theVL-CDR3comprisesanaminoacid sequencecorresponding toSEQID NO: 10 or 20. 30. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 816; wherein the VH-CDR3 comprises an amino acid sequence corre- sponding to SEQ IDNO: 15, andwherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. 31. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 818; wherein the VH-CDR3 comprises an amino acid sequence corre- sponding to SEQ IDNO: 15, andwherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20. 32. The antibody or antigen-binding fragment thereof of any one of aspects 1‑31, wherein the VH comprises framework regions that are at least 90% identical to each of SEQ ID NOs: 803‑806. 33. The antibody or antigen-binding fragment thereof of any one of aspects 1‑31, wherein the VH comprises framework regions that are at least 95% identical to each of SEQ ID NOs: 803‑806. 34. The antibody or antigen-binding fragment there of any one of aspects 1‑31, wherein the VH comprises framework regions corresponding to SEQ ID NOs: 803‑806. 35.Theantibodyorantigen-binding fragment thereof ofanyoneofaspects1‑34,wherein theVLcomprises framework regions that are at least 90% identical to each of SEQ ID NOs: 807‑810. 36.Theantibodyorantigen-binding fragment thereof ofanyoneofaspects1‑34,wherein theVLcomprises framework regions that are at least 95% identical to each of SEQ ID NOs: 807‑810. 37. The antibody or antigen-binding fragment there of any one of aspects 1‑34, wherein the VL comprises framework regions corresponding to SEQ ID NOs: 807‑810. 38. The antibody or antigen-binding fragment thereof of aspect 3, wherein theVHcomprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from the groupconsisting of: SEQ IDNOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792, and 821‑831. 39. The antibody or antigen-binding fragment thereof of aspect 3, wherein the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from the groupconsisting of: SEQ IDNOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 62 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797. 40. The antibody or antigen-binding fragment thereof of aspect 3, wherein theVHcomprises an amino acid sequence corresponding to SEQ ID NO: 821 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. 41. The antibody or antigen-binding fragment thereof of aspect 3, wherein theVHcomprises an amino acid sequence corresponding to SEQ ID NO: 824 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. 42. The antibody or antigen-binding fragment thereof of aspect 3, wherein theVHcomprises an amino acid sequence corresponding to SEQ ID NO: 827 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. 43. The antibody or antigen-binding fragment thereof of aspect 3, wherein theVHcomprises an amino acid sequence corresponding to SEQ ID NO: 831 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. 44. The antibody or antigen-binding fragment thereof of any one of aspects 3‑39, wherein theVHcomprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 12 and wherein the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 17. 45. The antibody or antigen-binding fragment of any one of aspects 1‑44, wherein the antibody or antigen-binding fragment is an antigen-binding fragment. 46. The antibody or antigen-binding fragment of aspect 45, wherein the antigen-binding fragment is an scFv. 47. The antibody or antigen-binding fragment of aspect 45, wherein the antigen-binding fragment is a Fab’. 48. The antibody or antigen-binding fragment of any one of aspects 1‑44, wherein the antibody or antigen-binding fragment is an antibody. 49. The antibody or antigen-binding fragment of aspect 48, wherein the antibody is a monoclonal antibody. 50. The antibody or antigen-binding fragment of aspect 48 or 49, wherein the antibody is an IgG antibody. 51. The antibody or antigen-binding fragment of any one of aspects 1‑50, wherein the antibody or antigen-binding fragment is humanized. 52. The antibody or antigen-binding fragment of any one of aspects 1‑50, wherein the antibody or antigen-binding fragment is human. 53. The antibody or antigen-binding fragment of any one of aspects 1‑52, wherein theVH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. 54. The antibody or antigen-binding fragment of aspect 53, wherein the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. 55.Theantibodyorantigen-binding fragmentof aspect53,wherein theVH isencodedbyanucleicacid comprising the nucleotide sequenceof any oneof SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 63 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. 56. The antibody or antigen-binding fragment of any one of aspects 1‑55, wherein the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. 57. The antibody or antigen-binding fragment of aspect 56, wherein the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. 58. Theantibodyor antigen-binding fragment of aspect 56,wherein theVL is encodedbyanucleic acid comprising the nucleotide sequenceof anyoneofSEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. 59. The antibody or antigen-binding fragment of any one of aspects 1‑58, wherein theVH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 11. 60. The antibody or antigen-binding fragment of aspect 59, wherein the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 11. 61.Theantibodyorantigen-binding fragmentof aspect59,wherein theVH isencodedbyanucleicacid comprising the nucleotide sequence of SEQ ID NO: 11. 62. The antibody or antigen-binding fragment of any one of aspects 1‑61, wherein the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 16. 63. The antibody or antigen-binding fragment of aspect 62, wherein the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 16. 64. Theantibodyor antigen-binding fragment of aspect 62,wherein theVL is encodedbyanucleic acid comprising the nucleotide sequence of SEQ ID NO: 16. 65. The antibody or antigen-binding fragment of any one of aspects 1‑64, wherein the antibody or antigen-binding fragment binds to PAR2. 66. The antibody or antigen-binding fragment of any one of aspects 1‑65, wherein the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from interacting with PAR2. 67. The antibody or antigen-binding fragment of any one of aspects 1‑66, wherein the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from cleaving PAR2. 68. The antibody or antigen-binding fragment of any one of aspects 1‑67, wherein the antibody or antigen-binding fragment prevents cleavage of the PAR2 extracellular domain. 69. The antibody or antigen-binding fragment of any one of aspects 1‑68, wherein the antibody or antigen-binding fragment inhibits exposure of the tethered ligand. 70. The antibody or antigen-binding fragment of any one of aspects 1‑69, wherein the antibody or antigen-binding fragment prevents the tethered ligand from interacting with the second transmembrane loop of PAR2. 64 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 71. The antibody or antigen-binding fragment of any one of aspects 1‑70, wherein the antibody or antigen-binding fragment binds to PAR2 with greater affinity at a pH of 7.4 than at a pH of 6.0. 72. The antibody or antigen-binding fragment of any one of aspects 1‑70, wherein the antibody or antigen-binding fragment bindsPAR2at pH7.4with aKDof less thanabout 5 nM, 1nM, 900pM, 800pM, 700pM,650pM,600pM, 500 pM, 200 pM, 100 pM or 50 pM. 73. The antibody or antigen-binding fragment of aspect 72, wherein the antibody or antigen-binding fragment binds PAR2 at pH 7.4 with a KD of less than about 700 pM. 74. The antibody or antigen-binding fragment of any one of aspects 1‑70, wherein the antibody or antigen-binding fragment bindsPAR2at pH6.0with aKDof greater than about 1 nM, 5 nM, 10 nM, 15 nM, 20 nM, 25nM, 30nM, 40nM, 50 nM, 60 nM, 80 nM, or 100 nM. 75. The antibody or antigen-binding fragment of any one of aspects 1‑70, wherein the antibody or antigen-binding fragment binds PAR2 at pH 6.0 with a KD of greater than about 30 nM. 76. The antibody or antigen-binding fragment of any one of aspects 1‑75, wherein the antibody or antigen-binding fragment has an IC50 of less than 100 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 7.4 in a PAR2 binding assay. 77. The antibody or antigen-binding fragment of any one of aspects 1‑76, wherein the antibody or antigen-binding fragment has an IC50 of less than 50 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 7.4 in a PAR2 binding assay. 78. The antibody or antigen-binding fragment of any one of aspects 1‑77, wherein the antibody or antigen-binding fragment has an IC50 of less than 40 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 7.4 in a PAR2 binding assay. 79. The antibody or antigen-binding fragment of any one of aspects 1‑78, wherein the antibody or antigen-binding fragment hasan IC50 of greater than500nMwhencompetingwith anantibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 6.0 in a PAR2 binding assay. 80. The antibody or antigen-binding fragment of any one of aspects 1‑79, wherein the antibody or antigen-binding fragment has an IC50 of greater than 1000 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 6.0 in a PAR2 binding assay. 81. The antibody or antigen-binding fragment of any one of aspects 1‑80, wherein the antibody or antigen-binding fragment has an IC50 of greater than 1100 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 6.0 in a PAR2 binding assay. 82. The antibody or antigen-binding fragment of any one of aspects 1‑81, wherein the antibody or antigen-binding fragment has an IC50 more than 20 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 in a PAR2 binding assay. 83. The antibody or antigen-binding fragment of any one of aspects 1‑82, wherein the antibody or antigen-binding fragment has an IC50 more than 25 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 in a PAR2 binding assay. 84. The antibody or antigen-binding fragment of any one of aspects 1‑83, wherein the antibody or antigen-binding fragment has an IC50 more than 30 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 in a PAR2 binding assay. 85. The antibody or antigen-binding fragment of any one of aspects 1‑84, wherein the antibody or antigen-binding fragment has an IC50 of less than 3.0 x 10‑10 M in a calcium influx assay in human A549 cells. 86. The antibody or antigen-binding fragment of any one of aspects 1‑85, wherein the antibody or antigen-binding fragment has an IC50 of less than 1.5 x 10‑10 M in a calcium influx assay in human A549 cells. 65 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 87. The antibody or antigen-binding fragment of any one of aspects 1‑86, wherein the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10‑10 M in a calcium influx assay in rat KNRK cells. 88. The antibody or antigen-binding fragment of any one of aspects 1‑87, wherein the antibody or antigen-binding fragment has an IC50 of less than 5.5 x 10‑10 M in a calcium influx assay in rat KNRK cells. 89. The antibody or antigen-binding fragment of any one of aspects 1‑88, wherein the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10‑11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells. 90. The antibody or antigen-binding fragment of any one of aspects 1‑89, wherein the antibody or antigen-binding fragment has an IC50 of less than 5.0 x 10‑11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells. 91. The antibody or antigen-binding fragment of any one of aspects 1‑90, wherein the antibody or antigen-binding fragment has an IC50 of less than 6.0 x 10‑11 M in a calcium influx assay in murine LL / 2 cells. 92. The antibody or antigen-binding fragment of any one of aspects 1‑91, wherein the antibody or antigen-binding fragment has an IC50 of less than 4.0 x 10‑11 M in a calcium influx assay in murine LL / 2 cells. 93. The antibody or antigen-binding fragment of any one of aspects 92, wherein the antibody or antigen-binding fragment is clearedmore slowly from serum of a treated patient than an antibody or antigen-binding fragment lacking histidine modifications. 94. The antibody or antigen-binding fragment of any one of aspects 3 and 5‑93, wherein the optionally present histidine or histidines reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by nomore than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having a VHwith an amino acid sequence of SEQ ID NO: 2 and VL with an amino acid sequence of SEQ ID NO: 7 when tested at a pH of 7.4 in a PAR2 binding assay. 95. A nucleic acid capable of expressing the antibody or antigen-binding fragment of any one of aspects 1‑94. 96. A nucleic acid comprising a nucleotide sequence that is at least 90% identical to any oneof SEQ IDNOs: 1, 11, 21, 31,41, 51,61, 71,81, 91,101, 111, 121,131, 141, 151, 161,171, 181, 191, 201,211, 221,231, 241, 251, 261,271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. 97. The nucleic acid of aspect 96, wherein the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. 98. The nucleic acid of aspect 96, wherein the nucleic acid comprises the nucleotide sequence of any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841. 99. A nucleic acid comprising anucleotide sequence that is at least 90% identical to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. 100. The nucleic acid of aspect 99, wherein the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 66 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. 101. The nucleic acid of aspect 99, wherein the nucleic acid comprises the nucleotide sequence of any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796. 102. A nucleic acid comprising a nucleotide sequence that is at least 90% identical to SEQ ID NO: 11. 103. The nucleic acid of aspect 102, wherein the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ ID NO: 11. 104. The nucleic acid of aspect 102, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 11. 105. A nucleic acid comprising a nucleotide sequence that is at least 90% identical to SEQ ID NO: 16. 106. The nucleic acid of aspect 105, wherein the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ ID NO: 16. 107. The nucleic acid of aspect 105, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 16. 108. A vector comprising the nucleic acid of any one of aspects 95‑107. 109. A set of vectors comprising: a) the nucleic acid of any one of aspects 96‑98 or 102‑104; and b) the nucleic acid of any one of aspects 99‑101 and 105‑107. 110. A host cell comprising one or more of the vectors of aspect 108 or 109. 111. A composition comprising a pharmaceutically acceptable carrier and the antibody or antigen-binding fragment of any one of aspects 1‑94. 112. A lyophilized composition comprising the antibody or antigen-binding fragment thereof of any one of aspects 1‑94. 113. A reconstituted lyophilized composition comprising the antibody or antigen-binding fragment thereof of any one of aspects 1‑94. 114. The composition of any one of aspects 111‑113, wherein the composition is formulated for administration by lozenge, spray, oral administration, delayed release or sustained release, transmucosal administration, syrup, mucoadhesive, buccal formulation, mucoadhesive tablet, topical administration, parenteral administration, injection, subdermal administration, oral solution, rectal administration, buccal administration or transdermal administration. 115. A kit comprising the antibody or antigen-binding fragment of any one of aspects 1‑94 or the composition of any one of aspects 111‑114. 116.Amethod for treatingpain in asubject in need thereof, comprisingadministering to thesubject apharmaceutically effective amount of the antibody or antigen-binding fragment of any one of aspects 1‑94. 117.Amethod for treatingpain in asubject in need thereof, comprisingadministering to thesubject apharmaceutically effective amount of the composition of any one of aspects 111‑114. 118. The method of aspect 116 or 117, wherein the pain is selected from the group consisting of: nociceptive, neuropathic, and mix-type pain. 119. Themethodof anyoneof aspects 116or 117,wherein thepain is associatedwith aheadache, chronic headache, a migraine headache, a cancer, a viral infection, rheumatoid arthritis, osteoarthritis, Crohn’s disease, liver disease, multiple sclerosis, spinal cord injury, post herpetic neuralgia, diabetic neuropathy, lower backpain, inflammatory heart 67 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 disease, kidney disease, gastritis, gingivitis, periodontal disease, asthma, chronic obstructive pulmonary disease, autoimmune disease, irritable bowel syndrome, fibromyalgia, leg pains, restless leg syndrome, diabetic neuropathy, an allergic condition, a surgical procedure, acute or chronic physical injury, bone fracture or a crush injury, spinal cord injury, an inflammatory disease, a non-inflammatory neuropathic or dysfunctional pain condition, or a combination thereof. 120. The method of aspect 119, wherein the pain is osteoarthritis pain. 121. The method of any one of aspects 116‑120, wherein the subject is a human. 122.Amethodof producing theantibodyor antigen-binding fragment of anyoneof aspects1‑94, comprising the steps of: expressing the nucleic acid of any one of aspects 95‑106 in a cultured cell, purifying the antibody or antigen-binding fragment. Claims 1. An antibody or antigen-binding fragment thereof comprising: (A) a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: (i)a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 3; (ii) a VH-CDR2 having the amino acid sequence of SEQ IDNO: 4, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 4; and (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 5; and wherein the VL comprises: (i)a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 8; (ii) a VL-CDR2 having the amino acid sequence of SEQ IDNO: 9, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 9; and (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 10; wherein the amino acid substitutions, deletions or insertions reduce the binding affinity of the antibody or antigen-binding fragment thereof for humanPAR2 by nomore than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having a VH with an amino acid sequence of SEQ ID NO: 2 and VL with an amino acid sequence of SEQ ID NO: 7 when tested at a pH of 7.4 in a PAR2 binding assay, optionallywherein theaminoacidsubstitutions, deletionsor insertionscomprisesahomologoussubstitution; or (B) a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: (i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3; (ii)a VH-CDR2having the amino acid sequence of SEQ IDNO: 4, butwherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑17 of SEQ ID NO: 4; and (iii) aVH-CDR3having theaminoacid sequenceofSEQ IDNO:5, butwherein ahistidine isoptionally present at any one or more of the amino acid positions corresponding to positions 1‑8 of SEQ ID NO: 5; 68 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 and wherein the VL comprises: (i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8; (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9; and (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1‑14 of SEQ ID NO: 10. 2. The antibody or antigen-binding fragment thereof of claim 1(B), wherein: (A) the VH comprises: (i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, (ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, and wherein the VL comprises: (i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8, (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9, (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; or (B) the VH comprises: (i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 13, (ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 14, (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 15, and wherein the VL comprises: (i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 18, (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 19, (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 20; or (C) a histidine is present at the amino acid positions corresponding to positions 5, 8, 12, 16, and 17 of SEQ IDNO: 4; andwherein a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ IDNO: 5. 3. The antibody or antigen-binding fragment thereof of claim 1(B) or claim 2, wherein: (A) a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 4; and / or (B) a histidine is present at the amino acid position corresponding to: (i) position 7 of SEQ ID NO: 4; or (ii) position 8 of SEQ ID NO: 4; or (iii) position 12 of SEQ ID NO: 4; and / or (C) a histidine is present at the amino acid position corresponding to position 15 of SEQ ID NO: 4; and / or (D) a histidine is present: (i) at the amino acid position corresponding to position 16 of SEQ ID NO: 4; or (ii) at the amino acid position corresponding to position 17 of SEQ ID NO: 4; or (iii) at the amino acid positions corresponding to positions 5 and 8 of SEQ ID NO: 4; or (iv) at the amino acid positions corresponding to positions 5, 8, 12, and 16 of SEQ ID NO: 4; or (v) at the amino acid positions corresponding to positions 5, 8, 12, 16 and 17 of SEQ ID NO: 4; and / or (E) a histidine is present at the amino acid position corresponding to position 2 of SEQ ID NO: 5; and / or (F) a histidine is present at the amino acid position corresponding to position 3 of SEQ ID NO: 5; and / or (G) a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ ID NO: 5; and / or 69 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 (H) a histidine is present at the amino acid position corresponding to position 1 of SEQ ID NO: 10; and / or (I) a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 10; and / or (J) a histidine is present at the amino acid position corresponding to position 6 of SEQ ID NO: 10; and / or (K) a histidine is present at the amino acid position corresponding to position 14 of SEQ ID NO: 10. 4. The antibody or antigen-binding fragment thereof of claim 1(B), wherein: (A) (i) the VH-CDR2 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794, and 811‑818, and / or (ii) the VH-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ IDNOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, and 819‑820; and / or (iii) theVL-CDR3comprises anaminoacid sequence selected from thegroup consisting of: SEQ IDNOs: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790 and 800; or (B) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 14; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 20; or (C) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 811; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 819, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (D) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 814; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 820, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (E) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 816; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (F) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 818; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (G) theVHcomprisesanaminoacidsequence that isat least80%,85%,90%,92%,93%,95%,97%,98%,99%or 100% identical to a sequence selected from the group consisting of: SEQ IDNOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792, and 821‑831; or (H) theVLcomprisesanaminoacid sequence that is at least 80%,85%,90%,92%,93%,95%,97%,98%,99%or 100% identical to a sequence selected from the group consisting of: SEQ IDNOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 70 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797; or (I) theVHcomprisesanaminoacidsequencecorresponding toSEQIDNO:821andwherein theVLcomprisesan amino acid sequence corresponding to SEQ ID NO: 7 or 17; or (J) the VH comprises an amino acid sequence corresponding to SEQ ID NO: 824 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17; or (K) the VH comprises an amino acid sequence corresponding to SEQ IDNO: 827 andwherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17; or (L) the VH comprises an amino acid sequence corresponding to SEQ IDNO: 831 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17. 5. The antibody or antigen-binding fragment thereof of any one of claims 1‑3 and 4(A)‑(G), wherein: (A) the VH comprises framework regions: (i) that are at least 90% identical to each of SEQ ID NOs: 803‑806; or (ii) that are at least 95% identical to each of SEQ ID NOs: 803‑806; or (iii) corresponding to SEQ ID NOs: 803‑806; and / or (B) the VL comprises framework regions: (i) that are at least 90% identical to each of SEQ ID NOs: 807‑810; or (ii) that are at least 95% identical to each of SEQ ID NOs: 807‑810; or (iii) corresponding to SEQ ID NOs: 807‑810. 6. The antibody or antigen-binding fragment thereof of any one of claims 1(B), 2, 3, 4(A)‑4(H) and 5, wherein the VH comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 12 and wherein the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 17. 7. The antibody or antigen-binding fragment of any one of claims 1‑6, wherein: (A) the antibody or antigen-binding fragment is: (i) an antigen-binding fragment, optionally wherein the antigen-binding fragment: (a) is an scFv; or (b) is a Fab’; or (ii) an antibody, optionally wherein: (a) the antibody is a monoclonal antibody; and / or (b) the antibody is an IgG antibody; and / or (B) the antibody or antigen-binding fragment is humanized or human; and / or (C) theVH isencodedbyanucleicacid comprisinganucleotidesequence that isat least 90% identity toanyoneof SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841, optionally wherein: (i) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any oneofSEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841; or (ii) the VH is encoded by a nucleic acid comprising the nucleotide sequence of any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 71 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841; and / or (D) theVL is encodedbyanucleic acid comprisinganucleotide sequence that is at least 90% identity to anyoneof SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796, optionally wherein (i) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any oneofSEQIDNOs:6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; or (ii) the VL is encoded by a nucleic acid comprising the nucleotide sequence of any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; and / or (E) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 11, optionally wherein (i) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 11; or (ii) the VH is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 11; and / or (F) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 16, optionally wherein (i) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 16; or (ii) the VL is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 16. 8. The antibody or antigen-binding fragment of any one of claims 1‑7, wherein: (A) the antibody or antigen-binding fragment binds to PAR2; and / or (B) the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from interacting with PAR2; and / or (C) the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from cleaving PAR2; and / or (D) the antibody or antigen-binding fragment prevents cleavage of the PAR2 extracellular domain; and / or (E) the antibody or antigen-binding fragment inhibits exposure of the tethered ligand; and / or (F) the antibody or antigen-binding fragment prevents the tethered ligand from interacting with the second transmembrane loop of PAR2; and / or (G) the antibody or antigen-binding fragment: (i) binds to PAR2 with greater affinity at a pH of 7.4 than at a pH of 6.0; or (ii) binds PAR2 at pH 7.4 with a KD of less than about 5 nM, 1 nM, 900 pM, 800 pM, 700 pM, 650 pM, 600 pM, 500pM,200pM,100pMor50pM,optionallywherein theantibodyorantigen-binding fragmentbindsPAR2at pH 7.4 with a KD of less than about 700 pM; or (iii) binds PAR2 at pH 6.0 with a KD of greater than about 1 nM, 5 nM, 10 nM, 15 nM, 20 nM, 25 nM, 30 nM, 40 nM, 50 nM, 60 nM, 80 nM, or 100 nM; or (iv) binds PAR2 at pH 6.0 with a KD of greater than about 30 nM; and / or (H) the antibody or antigen-binding fragment has an IC50 of less than 100 nMwhen competingwith an antibody or antigen-binding fragment having the CDRs of SEQ IDNOs: 3‑5 and 8‑10 at a pH of 7.4 in a PAR2 binding assay; 72 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 and / or (I) the antibody or antigen-binding fragment has an IC50 of less than 50 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ IDNOs: 3‑5 and 8‑10 at a pH of 7.4 in a PAR2 binding assay; and / or (J) the antibody or antigen-binding fragment has an IC50 of less than 40 nMwhen competing with an antibody or antigen-binding fragment having the CDRs of SEQ IDNOs: 3‑5 and 8‑10 at a pH of 7.4 in a PAR2 binding assay; and / or (K) theantibodyor antigen-binding fragment hasan IC50 of greater than500nMwhencompetingwithanantibody or antigen-binding fragment having theCDRsofSEQ IDNOs: 3‑5and8‑10at apHof 6.0 in aPAR2bindingassay; and / or (L) the antibody or antigen-binding fragment has an IC50 of greater than 1000 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 6.0 in a PAR2 binding assay; and / or (M) the antibody or antigen-binding fragment has an IC50 of greater than 1100 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3‑5 and 8‑10 at a pH of 6.0 in a PAR2 binding assay; and / or (N) theantibodyor antigen-binding fragment hasan IC50more than20 times lowerat apHof 7.4 thanat apHof6.0 when competingwith an antibody or antigen-binding fragment having theCDRsof SEQ IDNOs: 3‑5 and 8‑10 in a PAR2 binding assay; and / or (O) theantibodyorantigen-binding fragment hasan IC50more than25 times lowerat apHof7.4 thanatapHof6.0 when competingwith an antibody or antigen-binding fragment having theCDRsof SEQ IDNOs: 3‑5 and 8‑10 in a PAR2 binding assay; and / or (P) theantibodyor antigen-binding fragment hasan IC50more than30 times lowerat apHof 7.4 thanat apHof 6.0 when competingwith an antibody or antigen-binding fragment having theCDRsof SEQ IDNOs: 3‑5 and 8‑10 in a PAR2 binding assay; and / or (Q) the antibody or antigen-binding fragment has an IC50 of less than 3.0 x 10‑10 M in a calcium influx assay in human A549 cells; and / or (R) the antibody or antigen-binding fragment has an IC50 of less than 1.5 x 10‑10 M in a calcium influx assay in human A549 cells; and / or (S) the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10‑10 M in a calcium influx assay in rat KNRK cells; and / or (T) the antibody or antigen-binding fragment has an IC50 of less than 5.5 x 10‑10 M in a calcium influx assay in rat KNRK cells; and / or (U) the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10‑11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells; and / or (V) the antibody or antigen-binding fragment has an IC50 of less than 5.0 x 10‑11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells; and / or (W) the antibody or antigen-binding fragment has an IC50 of less than 6.0 x 10‑11 M in a calcium influx assay in murine LL / 2 cells; and / or (X) the antibody or antigen-binding fragment has an IC50 of less than 4.0 x 10‑11 M in a calcium influx assay in murine LL / 2 cells; and / or (Y) the antibody or antigen-binding fragment is cleared more slowly from serum of a treated patient than an antibody or antigen-binding fragment lacking histidine modifications. 9. The antibody or antigen-binding fragment of any one of claims 1(B) and 5‑93, wherein the optionally present histidine or histidines reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having a VHwith an amino acid sequence of SEQ ID NO: 2 and VL with an amino acid sequence of SEQ ID NO: 7 when tested at a pH of 7.4 in a PAR2 binding assay. 10. A nucleic acid: (A) capable of expressing the antibody or antigen-binding fragment of any one of claims 1‑9; or (B) comprisinganucleotide sequence that is at least 90% identical toanyoneofSEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841, 73 EP 4 681 726 A2 5 10 15 20 25 30 35 40 45 50 55 optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ IDNOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841; or (ii) thenucleicacid comprises thenucleotidesequenceofanyoneofSEQ IDNOs:1, 11,21, 31, 41,51, 61,71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833‑841; or (C) comprisinganucleotidesequence that is at least 90% identical toanyoneofSEQIDNOs:6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ IDNOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; or (ii) thenucleicacid comprises thenucleotidesequenceofanyoneofSEQIDNOs:6, 16,26,36,46,56,66,76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; or (D) comprising a nucleotide sequence that is at least 90% identical to SEQ ID NO: 11, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ ID NO: 11; or (ii) the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 11; or (E) comprising a nucleotide sequence that is at least 90% identical to SEQ ID NO: 16, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ ID NO: 16; or (ii) the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 16. 11. (A) A vector comprising the nucleic acid of claim 10; or (B) a set of vectors comprising: (i) the nucleic acid of claim 10(B) or claim 10(D); and (ii) the nucleic acid of claim 10(C) and claim 10(E). 12. A host cell comprising one or more of ...

Claims

1. An antibody or antigen-binding fragment thereof comprising: (A) a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: (i)a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 3; (ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 4; and (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 5; and wherein the VL comprises: (i)a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 8; (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9, but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 9; and (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; but wherein 1, 2, 3, 4, or 5 amino acid substitutions, deletions or insertions are optionally present in the sequence of SEQ ID NO: 10; wherein the amino acid substitutions, deletions or insertions reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having a VH with an amino acid sequence of SEQ ID NO: 2 and VL with an amino acid sequence of SEQ ID NO: 7 when tested at a pH of 7.4 in a PAR2 binding assay, optionally wherein the amino acid substitutions, deletions or insertions comprises a homologous substitution; or (B) a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH comprises: (i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3; (ii)a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1-17 of SEQ ID NO: 4; and (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1-8 of SEQ ID NO: 5; and wherein the VL comprises: (i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8; (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9; and (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; but wherein a histidine is optionally present at any one or more of the amino acid positions corresponding to positions 1-14 of SEQ ID NO: 10.

2. The antibody or antigen-binding fragment thereof of claim 1(B), wherein: (A) the VH comprises: (i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 3, (ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 4, (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 5, and wherein the VL comprises: (i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 8, (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 9, (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 10; or (B) the VH comprises: (i) a VH-CDR1 having the amino acid sequence of SEQ ID NO: 13, (ii) a VH-CDR2 having the amino acid sequence of SEQ ID NO: 14, (iii) a VH-CDR3 having the amino acid sequence of SEQ ID NO: 15, and wherein the VL comprises: (i) a VL-CDR1 having the amino acid sequence of SEQ ID NO: 18, (ii) a VL-CDR2 having the amino acid sequence of SEQ ID NO: 19, (iii) a VL-CDR3 having the amino acid sequence of SEQ ID NO: 20; or (C) a histidine is present at the amino acid positions corresponding to positions 5, 8, 12, 16, and 17 of SEQ ID NO: 4; and wherein a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ ID NO: 5.

3. The antibody or antigen-binding fragment thereof of claim 1(B) or claim 2, wherein: (A) a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 4; and / or (B) a histidine is present at the amino acid position corresponding to: (i) position 7 of SEQ ID NO: 4; or (ii) position 8 of SEQ ID NO: 4; or (iii) position 12 of SEQ ID NO: 4; and / or (C) a histidine is present at the amino acid position corresponding to position 15 of SEQ ID NO: 4; and / or (D) a histidine is present: (i) at the amino acid position corresponding to position 16 of SEQ ID NO: 4; or (ii) at the amino acid position corresponding to position 17 of SEQ ID NO: 4; or (iii) at the amino acid positions corresponding to positions 5 and 8 of SEQ ID NO: 4; or (iv) at the amino acid positions corresponding to positions 5, 8, 12, and 16 of SEQ ID NO: 4; or (v) at the amino acid positions corresponding to positions 5, 8, 12, 16 and 17 of SEQ ID NO: 4; and / or (E) a histidine is present at the amino acid position corresponding to position 2 of SEQ ID NO: 5; and / or (F) a histidine is present at the amino acid position corresponding to position 3 of SEQ ID NO: 5; and / or (G) a histidine is present at the amino acid positions corresponding to positions 2 and 3 of SEQ ID NO: 5; and / or (H) a histidine is present at the amino acid position corresponding to position 1 of SEQ ID NO: 10; and / or (I) a histidine is present at the amino acid position corresponding to position 5 of SEQ ID NO: 10; and / or (J) a histidine is present at the amino acid position corresponding to position 6 of SEQ ID NO: 10; and / or (K) a histidine is present at the amino acid position corresponding to position 14 of SEQ ID NO: 10.

4. The antibody or antigen-binding fragment thereof of claim 1(B), wherein: (A) (i) the VH-CDR2 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 4, 14, 24, 34, 44, 54, 64, 74, 84, 94, 104, 114, 124, 134, 144, 154, 164, 174, 184, 194, 204, 214, 224, 234, 244, 254, 264, 274, 284, 294, 304, 314, 324, 334, 344, 354, 364, 374, 384, 394, 404, 414, 424, 434, 444, 454, 464, 474, 484, 494, 504, 514, 524, 534, 544, 554, 564, 574, 584, 594, 604, 614, 624, 634, 644, 654, 664, 674, 684, 694, 704, 714, 724, 734, 744, 754, 764, 774, 784, 794, and 811-818, and / or (ii) the VH-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 5, 15, 25, 35, 45, 55, 65, 75, 85, 95, 105, 115, 125, 135, 145, 155, 165, 175, 185, 195, 205, 215, 225, 235, 245, 255, 265, 275, 285, 295, 305, 315, 325, 335, 345, 355, 365, 375, 385, 395, 405, 415, 425, 435, 445, 455, 465, 475, 485, 495, 505, 515, 525, 535, 545, 555, 565, 575, 585, 595, 605, 615, 625, 635, 645, 655, 665, 675, 685, 695, 705, 715, 725, 735, 745, 755, 765, 775, 785, 795, and 819-820; and / or (iii) the VL-CDR3 comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790 and 800; or (B) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 14; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 20; or (C) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 811; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 819, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (D) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 814; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 820, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (E) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 816; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (F) the VH-CDR2 comprises an amino acid sequence corresponding to SEQ ID NO: 818; wherein the VH-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 15, and wherein the VL-CDR3 comprises an amino acid sequence corresponding to SEQ ID NO: 10 or 20; or (G) the VH comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from the group consisting of: SEQ ID NOs: 2, 12, 22, 32, 42, 52, 62, 72, 82, 92, 102, 112, 122, 132, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402, 412, 422, 432, 442, 452, 462, 472, 482, 492, 502, 512, 522, 532, 542, 552, 562, 572, 582, 592, 602, 612, 622, 632, 642, 652, 662, 672, 682, 692, 702, 712, 722, 732, 742, 752, 762, 772, 782, 792, and 821-831; or (H) the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from the group consisting of: SEQ ID NOs: 7, 17, 27, 37, 47, 57, 67, 77, 87, 97, 107, 117, 127, 137, 147, 157, 167, 177, 187, 197, 207, 217, 227, 237, 247, 257, 267, 277, 287, 297, 307, 317, 327, 337, 347, 357, 367, 377, 387, 397, 407, 417, 427, 437, 447, 457, 467, 477, 487, 497, 507, 517, 527, 537, 547, 557, 567, 577, 587, 597, 607, 617, 627, 637, 647, 657, 667, 677, 687, 697, 707, 717, 727, 737, 747, 757, 767, 777, 787, and 797; or (I) the VH comprises an amino acid sequence corresponding to SEQ ID NO: 821 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17; or (J) the VH comprises an amino acid sequence corresponding to SEQ ID NO: 824 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17; or (K) the VH comprises an amino acid sequence corresponding to SEQ ID NO: 827 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17; or (L) the VH comprises an amino acid sequence corresponding to SEQ ID NO: 831 and wherein the VL comprises an amino acid sequence corresponding to SEQ ID NO: 7 or 17.

5. The antibody or antigen-binding fragment thereof of any one of claims 1-3 and 4(A)-(G), wherein: (A) the VH comprises framework regions: (i) that are at least 90% identical to each of SEQ ID NOs: 803-806; or (ii) that are at least 95% identical to each of SEQ ID NOs: 803-806; or (iii) corresponding to SEQ ID NOs: 803-806; and / or (B) the VL comprises framework regions: (i) that are at least 90% identical to each of SEQ ID NOs: 807-810; or (ii) that are at least 95% identical to each of SEQ ID NOs: 807-810; or (iii) corresponding to SEQ ID NOs: 807-810.

6. The antibody or antigen-binding fragment thereof of any one of claims 1(B), 2, 3, 4(A)-4(H) and 5, wherein the VH comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 12 and wherein the VL comprises an amino acid sequence that is at least 80%, 85%, 90%, 92%, 93%, 95%, 97%, 99% or 100% identical to SEQ ID NO: 17.

7. The antibody or antigen-binding fragment of any one of claims 1-6, wherein: (A) the antibody or antigen-binding fragment is: (i) an antigen-binding fragment, optionally wherein the antigen-binding fragment: (a) is an scFv; or (b) is a Fab'; or (ii) an antibody, optionally wherein: (a) the antibody is a monoclonal antibody; and / or (b) the antibody is an IgG antibody; and / or (B) the antibody or antigen-binding fragment is humanized or human; and / or (C) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833-841, optionally wherein: (i) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833-841; or (ii) the VH is encoded by a nucleic acid comprising the nucleotide sequence of any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833-841; and / or (D) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796, optionally wherein (i) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; or (ii) the VL is encoded by a nucleic acid comprising the nucleotide sequence of any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; and / or (E) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 11, optionally wherein (i) the VH is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 11; or (ii) the VH is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 11; and / or (F) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 90% identity to SEQ ID NO: 16, optionally wherein (i) the VL is encoded by a nucleic acid comprising a nucleotide sequence that is at least 95% identity to SEQ ID NO: 16; or (ii) the VL is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 16.

8. The antibody or antigen-binding fragment of any one of claims 1-7, wherein: (A) the antibody or antigen-binding fragment binds to PAR2; and / or (B) the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from interacting with PAR2; and / or (C) the antibody or antigen-binding fragment prevents trypsin, tryptase and / or matriptase from cleaving PAR2; and / or (D) the antibody or antigen-binding fragment prevents cleavage of the PAR2 extracellular domain; and / or (E) the antibody or antigen-binding fragment inhibits exposure of the tethered ligand; and / or (F) the antibody or antigen-binding fragment prevents the tethered ligand from interacting with the second transmembrane loop of PAR2; and / or (G) the antibody or antigen-binding fragment: (i) binds to PAR2 with greater affinity at a pH of 7.4 than at a pH of 6.0; or (ii) binds PAR2 at pH 7.4 with a KD of less than about 5 nM, 1 nM, 900 pM, 800 pM, 700 pM, 650 pM, 600 pM, 500 pM, 200 pM, 100 pM or 50 pM, optionally wherein the antibody or antigen-binding fragment binds PAR2 at pH 7.4 with a KD of less than about 700 pM; or (iii) binds PAR2 at pH 6.0 with a KD of greater than about 1 nM, 5 nM, 10 nM, 15 nM, 20 nM, 25 nM, 30 nM, 40 nM, 50 nM, 60 nM, 80 nM, or 100 nM; or (iv) binds PAR2 at pH 6.0 with a KD of greater than about 30 nM; and / or (H) the antibody or antigen-binding fragment has an IC50 of less than 100 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 at a pH of 7.4 in a PAR2 binding assay; and / or (I) the antibody or antigen-binding fragment has an IC50 of less than 50 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 at a pH of 7.4 in a PAR2 binding assay; and / or (J) the antibody or antigen-binding fragment has an IC50 of less than 40 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 at a pH of 7.4 in a PAR2 binding assay; and / or (K) the antibody or antigen-binding fragment has an IC50 of greater than 500 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 at a pH of 6.0 in a PAR2 binding assay; and / or (L) the antibody or antigen-binding fragment has an IC50 of greater than 1000 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 at a pH of 6.0 in a PAR2 binding assay; and / or (M) the antibody or antigen-binding fragment has an IC50 of greater than 1100 nM when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 at a pH of 6.0 in a PAR2 binding assay; and / or (N) the antibody or antigen-binding fragment has an IC50 more than 20 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 in a PAR2 binding assay; and / or (O) the antibody or antigen-binding fragment has an IC50 more than 25 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 in a PAR2 binding assay; and / or (P) the antibody or antigen-binding fragment has an IC50 more than 30 times lower at a pH of 7.4 than at a pH of 6.0 when competing with an antibody or antigen-binding fragment having the CDRs of SEQ ID NOs: 3-5 and 8-10 in a PAR2 binding assay; and / or (Q) the antibody or antigen-binding fragment has an IC50 of less than 3.0 x 10-10 M in a calcium influx assay in human A549 cells; and / or (R) the antibody or antigen-binding fragment has an IC50 of less than 1.5 x 10-10 M in a calcium influx assay in human A549 cells; and / or (S) the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10-10 M in a calcium influx assay in rat KNRK cells; and / or (T) the antibody or antigen-binding fragment has an IC50 of less than 5.5 x 10-10 M in a calcium influx assay in rat KNRK cells; and / or (U) the antibody or antigen-binding fragment has an IC50 of less than 7.0 x 10-11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells; and / or (V) the antibody or antigen-binding fragment has an IC50 of less than 5.0 x 10-11 M in a calcium influx assay in cynomolgus monkey CYNOM-K1 cells; and / or (W) the antibody or antigen-binding fragment has an IC50 of less than 6.0 x 10-11 M in a calcium influx assay in murine LL / 2 cells; and / or (X) the antibody or antigen-binding fragment has an IC50 of less than 4.0 x 10-11 M in a calcium influx assay in murine LL / 2 cells; and / or (Y) the antibody or antigen-binding fragment is cleared more slowly from serum of a treated patient than an antibody or antigen-binding fragment lacking histidine modifications.

9. The antibody or antigen-binding fragment of any one of claims 1(B) and 5-93, wherein the optionally present histidine or histidines reduce the binding affinity of the antibody or antigen-binding fragment thereof for human PAR2 by no more than 1000, 800, 700, 500, 400, 300, 200, 100, 50 or 10-fold as compared to an antibody or antigen-binding fragment having a VH with an amino acid sequence of SEQ ID NO: 2 and VL with an amino acid sequence of SEQ ID NO: 7 when tested at a pH of 7.4 in a PAR2 binding assay.

10. A nucleic acid: (A) capable of expressing the antibody or antigen-binding fragment of any one of claims 1-9; or (B) comprising a nucleotide sequence that is at least 90% identical to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833-841, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833-841; or (ii) the nucleic acid comprises the nucleotide sequence of any one of SEQ ID NOs: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121, 131, 141, 151, 161, 171, 181, 191, 201, 211, 221, 231, 241, 251, 261, 271, 281, 291, 301, 311, 321, 331, 341, 351, 361, 371, 381, 391, 401, 411, 421, 431, 441, 451, 461, 471, 481, 491, 501, 511, 521, 531, 541, 551, 561, 571, 581, 591, 601, 611, 621, 631, 641, 651, 661, 671, 681, 691, 701, 711, 721, 731, 741, 751, 761, 771, 781, 791, and 833-841; or (C) comprising a nucleotide sequence that is at least 90% identical to any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; or (ii) the nucleic acid comprises the nucleotide sequence of any one of SEQ ID NOs: 6, 16, 26, 36, 46, 56, 66, 76, 86, 96, 106, 116, 126, 136, 146, 156, 166, 176, 186, 196, 206, 216, 226, 236, 246, 256, 266, 276, 286, 296, 306, 316, 326, 336, 346, 356, 366, 376, 386, 396, 406, 416, 426, 436, 446, 456, 466, 476, 486, 496, 506, 516, 526, 536, 546, 556, 566, 576, 586, 596, 606, 616, 626, 636, 646, 656, 666, 676, 686, 696, 706, 716, 726, 736, 746, 756, 766, 776, 786, and 796; or (D) comprising a nucleotide sequence that is at least 90% identical to SEQ ID NO: 11, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ ID NO: 11; or (ii) the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 11; or (E) comprising a nucleotide sequence that is at least 90% identical to SEQ ID NO: 16, optionally wherein: (i) the nucleic acid comprises a nucleotide sequence that is at least 95% identical to SEQ ID NO: 16; or (ii) the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 16.

11. (A) A vector comprising the nucleic acid of claim 10; or (B) a set of vectors comprising: (i) the nucleic acid of claim 10(B) or claim 10(D); and (ii) the nucleic acid of claim 10(C) and claim 10(E).

12. A host cell comprising one or more of the vectors of claim 11.

13. A (A) composition, (B) lyophilized composition, or (C) reconstituted lyophilized composition comprising a pharmaceutically acceptable carrier and the antibody or antigen-binding fragment of any one of claims 1-9, optionally wherein the composition is formulated for administration by lozenge, spray, oral administration, delayed release or sustained release, transmucosal administration, syrup, mucoadhesive, buccal formulation, mucoadhesive tablet, topical administration, parenteral administration, injection, subdermal administration, oral solution, rectal administration, buccal administration or transdermal administration.

14. The antibody or antigen-binding fragment of any one of claims 1-9 or the composition of claim 13 for use in a method for treating pain in a subject in need thereof, comprising administering to the subject a pharmaceutically effective amount of the antibody or antigen-binding fragment of any one of claims 1-9 or the composition of claim 13, optionally wherein: (A) the pain is: (i) selected from the group consisting of: nociceptive, neuropathic, and mix-type pain; or (ii) associated with a headache, chronic headache, a migraine headache, a cancer, a viral infection, rheumatoid arthritis, osteoarthritis, Crohn's disease, liver disease, multiple sclerosis, spinal cord injury, post herpetic neuralgia, diabetic neuropathy, lower back pain, inflammatory heart disease, kidney disease, gastritis, gingivitis, periodontal disease, asthma, chronic obstructive pulmonary disease, autoimmune disease, irritable bowel syndrome, fibromyalgia, leg pains, restless leg syndrome, diabetic neuropathy, an allergic condition, a surgical procedure, acute or chronic physical injury, bone fracture or a crush injury, spinal cord injury, an inflammatory disease, a non-inflammatory neuropathic or dysfunctional pain condition, or a combination thereof, further optionally wherein the pain is osteoarthritis pain; and / or (B) the subject is a human.

15. A method of producing the antibody or antigen-binding fragment of any one of claims 1-9, comprising the steps of: expressing the nucleic acid of claim 10 in a cultured cell, purifying the antibody or antigen-binding fragment.

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