CN117120089A - Treatment of chronic prurigo - Google Patents

Treatment of chronic prurigo Download PDF

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Publication number
CN117120089A
CN117120089A CN202280023482.5A CN202280023482A CN117120089A CN 117120089 A CN117120089 A CN 117120089A CN 202280023482 A CN202280023482 A CN 202280023482A CN 117120089 A CN117120089 A CN 117120089A
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amino acid
seq
antibody
kit
antigen
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Chinese (zh)
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布雷特·艾伦·霍利
J·戈尔茨坦
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Celldex Therapeutics Inc
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Celldex Therapeutics Inc
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Priority claimed from PCT/US2022/013380 external-priority patent/WO2022159744A1/en
Publication of CN117120089A publication Critical patent/CN117120089A/en
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Abstract

Provided herein are methods and uses of inhibitors of KIT (receptor tyrosine kinase), e.g., antibodies and antigen binding fragments that immunospecifically bind to KIT, for controlling, treating, or preventing chronic prurigo, including Prurigo Nodularis (PN). Improved antibodies for use in these methods and uses are also provided.

Description

Treatment of chronic prurigo
RELATED APPLICATIONS
The present application claims the benefit of U.S. provisional patent application Ser. No.63/140,621, filed on 22. 1/2021, and 63/238,688 filed on 30. 8/2021, which are incorporated herein by reference in their entireties.
Reference to an electronically submitted sequence Listing
The present application incorporates by reference the sequence listing filed with the present application as a text file entitled "seqlipsting_12638-171-228. Txt" created at 1/6/2022 and having a size of 44,163 bytes.
1. Technical field
Provided herein are methods and uses of inhibitors of KIT (receptor tyrosine kinase), e.g., antibodies and antigen binding fragments that immunospecifically bind to KIT, for controlling, treating, or preventing chronic prurigo, including Prurigo Nodularis (PN). Improved antibodies for use in these methods and uses are also provided.
2. Background art
KIT (or c-KIT) is a type III receptor tyrosine kinase encoded by the c-KIT gene. KIT comprises 5 extracellular immunoglobulin (Ig) -like domains separated by a kinase insert, a single transmembrane region, an inhibitory cytoplasmic membrane-proximal domain, and a cytokinin kinase domain (see, e.g., yarden et al, nature,1986,323:226-232;Ullrich and Schlessinger,Cell,1990,61:203-212; clifford et al, j. Biol. Chem.,2003,278: 31461-31464). The human c-KIT gene encoding the KIT receptor has been cloned as described by Yarden et al, EMBO J.,1987, 6:3341-3351. KIT is also known as CD117 or stem cell factor receptor ("SCFR") because it is a receptor for stem cell factor ("SCF") ligands (also known as Steel factor or KIT ligand). SCF ligands that bind to the first three extracellular Ig-like domains of KIT cause receptor dimerization and thereby activate endogenous tyrosine kinase activity through phosphorylation of specific tyrosine residues in the juxtamembrane and kinase domains (see, e.g., weiss and Schlessinger, cell,1998,94:277-280; clifford et al, j. Biol. Chem.,2003, 278:31461-31464). Stat, src, ERK and AKT signaling pathway members have been shown to be downstream signaling proteins of KIT signaling.
The fourth (D4) and fifth (D5) extracellular Ig-like domains of KIT are believed to mediate receptor dimerization (see, e.g., international patent application publication No. WO 2008/153926; yuzawa et al, cell,2007, 130:323-334).
Expression of KIT has been detected in a variety of cell types, such as mast cells, stem cells, brain cells, melanocytes, ovarian cells, and cancer cells (e.g., leukemia cells) (see, e.g., prism, p.curr.opin.cell Biol,1991,3:939-946; lyman et al, blood,1998,91:1101-1134; ashman, l.k., int.j. Biochem.cell Biol,1999,31:1037-1051; kitamura et al, mutat.Res.,2001,477:165-171; mol et al, j.biol.chem.,2003, 278:31461-31464). In addition, KIT plays an important role in hematopoiesis, melanogenesis and gametogenesis (see Ueda et al, blood,2002, 99:3342-3349).
Antibodies against human KIT are known, for example, from international patent publication No WO2014018625A1, the disclosure of which is incorporated herein by reference in its entirety.
Chronic Prurigo (CPG) is a disease characterized by chronic itching (itching feel) and the presence of multiple local or systemic prurigo skin lesions.
Prurigo nodularis (Prurigo nodularis, PN; also known as prurigo nodularis) is a different clinical disease defined by chronic itching and the presence of multiple local or systemic, protruding, hard and nodular skin lesions. Although the underlying cause of PN is unknown, both neurological and immunological processes appear to play a role in their development (Elmariah et al, practical approaches for diagnosis and management of prurigo nodularis: US expert panel consensus, journal of the American Academy of Dermatology,2020, S0190-9622 (20): 32189-32187). Also refer to Periera et al, journal of the European Academy of Dermatology and Venereology (JEADV), 2018,32:1059-1065 and Zeidler et al, acta Derm Venereol,2018,98:173-179.
There is a need for effective control or treatment of chronic prurigo, including prurigo nodularis, and for providing therapies for improved antibodies for use in such treatment.
3. Summary of the invention
In one aspect, provided herein is a method of protecting, treating, or controlling chronic prurigo in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT.
In another aspect, provided herein is the use of an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT for the manufacture of a medicament for protecting, treating, or controlling chronic prurigo in a subject.
In another aspect, provided herein are antibodies or antigen-binding fragments thereof that immunospecifically bind to human KIT for use in a method of protecting, treating, or controlling chronic prurigo in a subject.
In a specific embodiment, the human KIT comprises SEQ ID NO:1, and a polypeptide having the amino acid sequence shown in 1.
In a specific embodiment, the chronic prurigo is prurigo nodularis.
In a specific embodiment, the antibody is a bivalent monospecific antibody. In a specific embodiment, the antibody is a bispecific antibody.
In a specific embodiment, the antibody is a humanized antibody.
In particular embodiments, the antibodies comprise a modified (e.g., mutated) Fc region or domain.
In specific embodiments, the antibody has reduced Fc receptor binding activity (specifically reduced fcγr binding activity).
In particular embodiments, the antibody does not cause significant degranulation of FcgRI-expressing human mast cells.
In particular embodiments, the antibody does not exhibit significant Fc receptor-dependent KIT agonist activity.
In specific embodiments, the antibody specifically binds to the D4 or D5 region of human KIT.
In particular embodiments, the antibody comprises (a) (i) a light chain variable region ("VL") comprising a sequence comprising SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and (ii) a heavy chain variable region ("VH") comprising the amino acid sequences comprising SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown; (B) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and (ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:26 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no; (C) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 28. SEQ ID NO:29 and SEQ ID NO:30, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and (ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:31 and SEQ ID NO:32, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown; (D) (i) VL comprising sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and (ii) VH comprising amino acid sequences comprising SEQ ID NOs: 33. SEQ ID NO:34 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no; or (E) VL comprising a sequence comprising SEQ ID NO: 35. SEQ ID NO:36 and SEQ ID NO:37, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and (ii) VH comprising amino acid sequences comprising SEQ ID NOs: 38. SEQ ID NO:39 and SEQ ID NO:40, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown.
In particular embodiments, the antibody comprises a light chain variable region ("VL") comprising a sequence comprising SEQ ID NO:2-4, and a heavy chain variable region ("VH") comprising the amino acid sequences set forth in SEQ ID NOs: 5-7, and VH CDR 1-3 of the amino acid sequence shown.
In a specific embodiment, the antibody comprises (i) a VL comprising the amino acid sequence: DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSASYR YSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVEIK (SEQ ID NO: 17), wherein X K1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, X K2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, X K3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is provided with aliphatic hydroxyl groupsAmino acids of the side chains of radicals or P, X K5 Is an amino acid having a charged or acidic side chain and X K6 Is an amino acid having an aromatic side chain; and (ii) VH comprising the amino acid sequence: QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIARIY PGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGVYY FDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain. In a specific embodiment, X K1 Is amino acid F or S, X K2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T, X K6 Is amino acid F or Y, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
In a specific embodiment, the antibody comprises a VL comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 13. 14, 15 and 16; and VH comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 8. 9, 10, 11 and 12.
In specific embodiments, the antibody comprises a human light chain constant region. In specific embodiments, the antibody comprises a human heavy chain constant region. In a specific embodiment, the human heavy chain constant region is a human IgG constant region. In a specific embodiment, the human heavy chain constant region is a human IgG1 constant region.
In particular embodiments, the antibodies comprise a modified (e.g., mutated) human Fc region or domain. In particular embodiments, the antibodies comprise a modified (e.g., mutated) human IgG1 Fc region or domain. In particular embodiments, the modified (e.g., mutated) human IgG1 Fc region or domain comprises the non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat. In particular embodiments, the modified (e.g., mutated) human IgG1 Fc region or domain further comprises the non-naturally occurring amino acids 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
In a specific embodiment, the antibody comprises: (i) a polypeptide comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no; (ii) a polypeptide comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat.
In a specific embodiment, the antibody comprises: (i) a polypeptide comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no; (ii) a polypeptide comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
In specific embodiments, the antibody comprises a heavy chain comprising the amino acid sequence: QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21).
In specific embodiments, the antibody comprises a light chain comprising the amino acid sequence: DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 22).
In specific embodiments, the antibody comprises a heavy chain comprising the amino acid sequence: QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21); and a light chain comprising the amino acid sequence: DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 22).
In a specific embodiment, the subject is an adult. In particular embodiments, the subject is a child.
3.1 illustrative embodiments
3.1.1 group 1
1. A method of protecting, treating, or controlling chronic prurigo in a subject comprising administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT.
2. The method of embodiment 1, wherein the human KIT comprises SEQ ID NO:1, and a polypeptide having the amino acid sequence shown in 1.
3. The method of embodiment 1 or 2, wherein the chronic prurigo is prurigo nodularis.
4. The method of any one of embodiments 1 to 3, wherein the antibody is a bivalent monospecific antibody.
5. The method of any one of embodiments 1 to 3, wherein the antibody is a bispecific antibody.
6. The method of any one of embodiments 1 to 5, wherein the antibody is a humanized antibody.
7. The method of any one of embodiments 1 to 6, wherein the antibody comprises a modified (e.g., mutated) Fc region or domain.
8. The method of any one of embodiments 1-7, wherein the antibody has reduced Fc receptor binding activity.
9. The method of any one of embodiments 1 to 8, wherein the antibody has reduced fcγr binding activity.
10. The method of any one of embodiments 1-9, wherein the antibody does not cause significant degranulation of FcgRI-expressing human mast cells.
11. The method of any one of embodiments 1-10, wherein the antibody does not exhibit significant Fc receptor-dependent KIT agonist activity.
12. The method of any one of embodiments 1-11, wherein the antibody specifically binds to the D4 or D5 region of human KIT.
13. The method of any one of embodiments 1 to 12, wherein the antibody comprises:
(A) (i) a light chain variable region ("VL") comprising a sequence comprising SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) A heavy chain variable region ("VH") comprising the amino acid sequences comprising SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown;
(B) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:26 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no;
(C) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 28. SEQ ID NO:29 and SEQ ID NO:30, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:31 and SEQ ID NO:32, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown;
(D) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 33. SEQ ID NO:34 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no; or alternatively
(E) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 35. SEQ ID NO:36 and SEQ ID NO:37, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 38. SEQ ID NO:39 and SEQ ID NO:40, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown.
14. The method of embodiment 13, wherein the antibody comprises a VL comprising a sequence comprising SEQ ID NO:2-4, said VH comprising VL CDRs 1-3 comprising the amino acid sequences set forth in SEQ ID NOs: 5-7, and VH CDR 1-3 of the amino acid sequence shown.
15. The method of any one of embodiments 1 to 14, wherein the antibody comprises
(i) VL comprising the amino acid sequence:
DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSAS YRYSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVE IK (SEQ ID NO: 17), wherein X K1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, X K2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, X K3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Is an amino acid having a charged or acidic side chain and X K6 Is an amino acid having an aromatic side chain; and
(ii) VH comprising the amino acid sequence:
QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIA RIYPGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGV YYFDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain.
16. The method of embodiment 15, wherein X K1 Is amino acid F or S, X K2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T, X K6 Is amino acid F or Y, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
17. The method of any one of embodiments 1 to 16, wherein the antibody comprises a VL comprising an amino acid sequence selected from the group consisting of seq id nos: SEQ ID NO: 13. 14, 15 and 16; and VH comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 8. 9, 10, 11 and 12.
18. The method of any one of embodiments 1 to 17, wherein the antibody comprises a human light chain constant region.
19. The method of any one of embodiments 1 to 18, wherein the antibody comprises a human heavy chain constant region.
20. The method of embodiment 19, wherein the human heavy chain constant region is a human IgG constant region.
21. The method of embodiment 20, wherein the human heavy chain constant region is a human IgG1 constant region.
22. The method of any one of embodiments 1 to 21, wherein the antibody comprises a modified (e.g., mutated) human Fc region or domain.
23. The method of any one of embodiments 1 to 21, wherein the antibody comprises a modified (e.g., mutated) human IgG1 Fc region or domain.
24. The method of embodiment 23, wherein the modified (e.g., mutated) human IgG1 Fc region or domain comprises the non-naturally occurring amino acids 234A, 235Q, and 322Q as numbered by the EU index as set forth in Kabat.
25. The method of embodiment 24, wherein the modified (e.g., mutated) human IgG1 Fc region or domain further comprises non-naturally occurring amino acids 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
26. The method of any one of embodiments 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat.
27. The method of any one of embodiments 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified (e.g., mutated) human IgG1 Fc region or domain comprising the non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
28. The method of any one of embodiments 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO：21)。
29. the method of any one of embodiments 1 to 25, wherein the antibody comprises a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
30. the method of any one of embodiments 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21); and a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
31. the method of any one of embodiments 1-30, wherein the subject is an adult.
32. The method of any one of embodiments 1 to 30, wherein the subject is a child.
3.1.2 group 2
1. Use of an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT for the manufacture of a medicament for protecting, treating or controlling chronic prurigo in a subject.
2. The use of embodiment 1, wherein the human KIT comprises SEQ ID NO:1, and a polypeptide having the amino acid sequence shown in 1.
3. The use of embodiment 1 or 2, wherein the chronic prurigo is prurigo nodularis.
4. The use according to any one of embodiments 1 to 3, wherein the antibody is a bivalent monospecific antibody.
5. The use according to any one of embodiments 1 to 3, wherein the antibody is a bispecific antibody.
6. The use according to any one of embodiments 1 to 5, wherein the antibody is a humanized antibody.
7. The use of any one of embodiments 1 to 6, wherein the antibody comprises a modified (e.g., mutated) Fc region or domain.
8. The use of any one of embodiments 1 to 7, wherein the antibody has reduced Fc receptor binding activity.
9. The use of any one of embodiments 1 to 8, wherein the antibody has reduced fcγr binding activity.
10. The use of any one of embodiments 1 to 9, wherein the antibody does not cause significant degranulation of FcgRI-expressing human mast cells.
11. The use of any one of embodiments 1-10, wherein the antibody does not exhibit significant Fc receptor-dependent KIT agonist activity.
12. The use of any one of embodiments 1 to 11, wherein the antibody specifically binds to the D4 or D5 region of human KIT.
13. The use according to any one of embodiments 1 to 12, wherein the antibody comprises:
(A) (i) a light chain variable region ("VL") comprising a sequence comprising SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) A heavy chain variable region ("VH") comprising the amino acid sequences comprising SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown;
(B) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:26 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no;
(C) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 28. SEQ ID NO:29 and SEQ ID NO:30, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:31 and SEQ ID NO:32, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown;
(D) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 33. SEQ ID NO:34 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no; or alternatively
(E) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 35. SEQ ID NO:36 and SEQ ID NO:37, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 38. SEQ ID NO:39 and SEQ ID NO:40, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown.
14. The use of any one of embodiments 1 to 13, wherein the antibody comprises a VL comprising a sequence comprising SEQ ID NO:2-4, said VH comprising VL CDRs 1-3 comprising the amino acid sequences set forth in SEQ ID NOs: 5-7, and VH CDR 1-3 of the amino acid sequence shown.
15. The use according to any one of embodiments 1 to 13, wherein the antibody comprises
(i) VL comprising the amino acid sequence:
DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSAS YRYSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVE IK (SEQ ID NO: 17), wherein X K1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, X K2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, X K3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Is an amino acid having a charged or acidic side chain and X K6 Is an amino acid having an aromatic side chain; and
(ii) VH comprising the amino acid sequence:
QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIA RIYPGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGV YYFDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain.
16. The use according to embodiment 15, wherein X K1 Is amino acid F or S, X K2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T, X K6 Is amino acid F or Y, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
17. The use of any one of embodiments 1 to 16, wherein the antibody comprises a VL comprising an amino acid sequence selected from the group consisting of seq id nos: SEQ ID NO: 13. 14, 15 and 16; and VH comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 8. 9, 10, 11 and 12.
18. The use of any one of embodiments 1 to 17, wherein the antibody comprises a human light chain constant region.
19. The use of any one of embodiments 1 to 18, wherein the antibody comprises a human heavy chain constant region.
20. The use of embodiment 19, wherein the human heavy chain constant region is a human IgG constant region.
21. The use of embodiment 20, wherein the human heavy chain constant region is a human IgG1 constant region.
22. The use of any one of embodiments 1 to 21, wherein the antibody comprises a modified (e.g., mutated) human Fc region or domain.
23. The use of any one of embodiments 1 to 21, wherein the antibody comprises a modified (e.g., mutated) human IgG1 Fc region or domain.
24. The use of embodiment 23, wherein the modified (e.g., mutated) human IgG1 Fc region or domain comprises the non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat.
25. The use of embodiment 24, wherein the modified (e.g., mutated) human IgG1 Fc region or domain further comprises non-naturally occurring amino acids 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
26. The use of any one of embodiments 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat.
27. The use of any one of embodiments 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified (e.g., mutated) human IgG1 Fc region or domain comprising the non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
28. The use according to any one of embodiments 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO：21)。
29. the use according to any one of embodiments 1 to 25, wherein the antibody comprises a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
30. the use according to any one of embodiments 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21); and a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
31. the use of any one of embodiments 1 to 30, wherein the subject is an adult.
32. The use of any one of embodiments 1 to 30, wherein the subject is a child.
3.1.3 group 3
1. An antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT for use in a method of protecting, treating or controlling chronic prurigo in a subject.
2. An antibody or antigen-binding fragment thereof for use according to embodiment 1, wherein the human KIT comprises the amino acid sequence of SEQ ID NO:1, and a polypeptide having the amino acid sequence shown in 1.
3. An antibody or antigen-binding fragment thereof for use according to embodiments 1 or 2, wherein the chronic prurigo is prurigo nodularis.
4. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 3, wherein the antibody is a bivalent monospecific antibody.
5. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 3, wherein the antibody is a bispecific antibody.
6. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 5, wherein the antibody is a humanized antibody.
7. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 6, wherein the antibody comprises a modified (e.g., mutated) Fc region or domain.
8. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 7, wherein the antibody has reduced Fc receptor binding activity.
9. An antibody or antigen binding fragment thereof for use according to any one of embodiments 1 to 8, wherein the antibody has reduced fcγr binding activity.
10. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 9, wherein the antibody does not cause significant degranulation of FcgRI-expressing human mast cells.
11. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 10, wherein the antibody does not exhibit significant Fc receptor-dependent KIT agonist activity.
12. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 11, wherein the antibody specifically binds to the D4 or D5 region of human KIT.
13. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 12, wherein the antibody comprises:
(A) (i) a light chain variable region ("VL") comprising a sequence comprising SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) A heavy chain variable region ("VH") comprising the amino acid sequences comprising SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown;
(B) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:26 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no;
(C) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 28. SEQ ID NO:29 and SEQ ID NO:30, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:31 and SEQ ID NO:32, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown;
(D) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 33. SEQ ID NO:34 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no; or alternatively
(E) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 35. SEQ ID NO:36 and SEQ ID NO:37, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 38. SEQ ID NO:39 and SEQ ID NO:40, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown.
14. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 13, wherein the antibody comprises a VL comprising a sequence comprising SEQ ID NO:2-4, said VH comprising VL CDRs 1-3 comprising the amino acid sequences set forth in SEQ ID NOs: 5-7, and VH CDR 1-3 of the amino acid sequence shown.
15. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 14, wherein the antibody comprises
(i) VL comprising the amino acid sequence:
DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSAS YRYSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVE IK (SEQ ID NO: 17), wherein X K1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, X K2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, X K3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Is an amino acid having a charged or acidic side chain and X K6 Is an amino acid having an aromatic side chain; and
(ii) VH comprising the amino acid sequence:
QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIA RIYPGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGV YYFDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain.
16. An antibody or antigen-binding fragment thereof for use according to embodiment 15, wherein X K1 Is amino acid F or S, X K2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T, X K6 Is amino acid F or Y, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
17. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 16, wherein the antibody comprises a VL comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 13. 14, 15 and 16; and VH comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 8. 9, 10, 11 and 12.
18. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 17, wherein the antibody comprises a human light chain constant region.
19. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 18, wherein the antibody comprises a human heavy chain constant region.
20. The antibody or antigen-binding fragment thereof for use according to embodiment 19, wherein the human heavy chain constant region is a human IgG constant region.
21. An antibody or antigen-binding fragment thereof for use according to embodiment 20, wherein the human heavy chain constant region is a human IgG1 constant region.
22. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 21, wherein the antibody comprises a modified (e.g., mutated) human Fc region or domain.
23. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 21, wherein the antibody comprises a modified (e.g., mutated) human IgG1 Fc region or domain.
24. An antibody or antigen-binding fragment thereof for use according to embodiment 23, wherein the modified (e.g., mutated) human IgG1 Fc region or domain comprises the non-naturally occurring amino acids 234A, 235Q and 322Q as numbered by the EU index as set forth in Kabat.
25. An antibody or antigen-binding fragment thereof for use according to embodiment 24, wherein the modified (e.g., mutated) human IgG1 Fc region or domain further comprises the non-naturally occurring amino acids 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
26. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat.
27. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified (e.g., mutated) human IgG1 Fc region or domain comprising the non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
28. An antibody or antigen binding fragment thereof for use according to any one of embodiments 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO：21)。
29. an antibody or antigen binding fragment thereof for use according to any one of embodiments 1 to 25, wherein the antibody comprises a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
30. An antibody or antigen binding fragment thereof for use according to any one of embodiments 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21); and a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
31. an antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 30, wherein the subject is an adult.
32. An antibody or antigen-binding fragment thereof for use according to any one of embodiments 1 to 30, wherein the subject is a child.
4. Description of the drawings
FIG. 1 shows full-length human KIT (SEQ ID NO: 1), genBank TM Amino acid sequence of accession number AAC 50969. First through fifth extracellular Ig-like domains (i.e., D1, D2, D3, D4, and D5) are shown; "{" shows the amino-terminal residue of each domain and "}" shows the carboxy-terminal residue of each domain. The D1 domain is shown at P34 to R112, the D2 domain is shown at D113 to P206, the D3 domain is shown at a207 to D309, the D4 domain is shown at K310 to N410, the hinge region between D4 and D5 is located at V409 to N410, and the D5 domain is shown at T411 to K509. In addition, the D1/D2 hinge region is located at D113 to L117; the D2/D3 hinge region is located between P206 and A210; and the D3/D4 hinge regions are located at D309 to G311. The D4/D5 region contains K310 through K509. The transmembrane domain comprises residues F525 to Q545 and the kinase domain comprises residues K589 to S933.
FIGS. 2A-2E show the effect of a particular anti-KIT antibody mAb1 on plasma tryptase levels according to the invention.
FIGS. 3A and 3B further illustrate the effect of a particular anti-KIT antibody mAb1 on plasma tryptase levels according to the invention.
FIG. 4 shows the effect of a particular anti-KIT antibody mAb1 on plasma Stem Cell Factor (SCF) levels according to the invention.
FIG. 5 shows the effect of a specific anti-KIT antibody mAb1 according to the invention and a corresponding antibody mAbc having the same variable region sequence but not mutated (wild-type) human IgG1 sequence on SCF-induced activation of wild-type KIT and downstream intracellular signaling pathways.
FIG. 6 shows the effect of a specific anti-KIT antibody mAb1 according to the invention and a corresponding antibody mAbc having the same variable domain sequence but the unmutated (wild-type) human IgG1 sequence on SCF-dependent cell proliferation.
FIG. 7 shows the binding affinities of a specific anti-KIT antibody mAb1 according to the invention and a corresponding antibody mAbc having the same variable domain sequence but the unmutated (wild-type) human IgG1 sequence to recombinant human Fc-gamma receptor (Fc gamma R) and human neonatal Fc receptor (FcRn).
Figures 8A-8N show the binding curves of a specific anti-KIT antibody mAb1 according to the invention and a corresponding antibody mAbc having the same variable region sequence but not mutated (wild-type) human IgG1 sequence to recombinant human Fc-gamma receptor (fcγr) and human neonatal Fc receptor (FcRn). Fig. 8A shows the binding curve of mAb1 to fcyri. Fig. 8B shows the binding curve of mAb1 to fcyriia. Fig. 8C shows the binding curve of mAb1 to fcyriib. Fig. 8D shows the binding curve of mAb1 to fcγriiia. Fig. 8E shows the binding curve of mAb1 to fcyriiib. Figure 8F shows the binding curve of mAb1 to FcRn (pH 6.0). FIG. 8G shows the binding curve of mAb1 to FcRn (pH 7.2). Fig. 8H shows the binding curve of mAbc to fcyri. Fig. 8I shows the binding curve of mAbc to fcyriia. Fig. 8J shows the binding curve of mAbc to fcyriib. Fig. 8K shows mAbc versus fcγriiia binding curve. Fig. 8L shows the binding curve of mAbc to fcγriiib. Figure 8M shows the binding curve of mAbc to FcRn (pH 6.0). Figure 8N shows the binding curve of mAbc to FcRn (pH 7.2).
FIG. 9 shows the effect of a particular anti-KIT antibody mAb1 according to the invention and a corresponding antibody mAbc having the same variable region sequence but an unmutated (wild-type) human IgG1 sequence on antibody-dependent cell-mediated cytotoxicity (ADCC) activity.
FIG. 10 shows the effect of a particular anti-KIT antibody mAb1 on specific cytokine production according to the invention. The conditions shown for each histogram are from left to right: PHA, LPS, huIgG1 (soluble), mAb 1.02 nM (soluble), mAb 1.2 nM (soluble), mAb 1.40 nM (soluble), mAb 1.02 nM (dry-coated), mAb 1.2 nM (dry-coated) and mAb 1.40 nM (dry-coated).
Figure 11 shows a schematic showing the effect of KIT signal transduction in mast cells and the effect of mAb1 on KIT receptors.
Figures 12A-12D show that a single dose of mAb1 resulted in a rapid and sustained response, with a Complete Response (CR) rate of 95% in patients with chronic induced urticaria (CIndU). 10/10 cold urticaria (Cold U) patients achieved CR (FIG. 12A). 8/9 symptomatic skin Scarification (SD) patients achieved CR and 1/9SD patients achieved Partial Response (PR) (FIG. 12B). Cr=negative challenge test at ∈4 ℃ or 0 needle; pr=improvement by 4 ℃ or ≡2 needles; the maximum response per patient is shown. FIG. 12C shows time course in a ColdU patient As a result. In a complete cold patient (n=8), CR was maintained for a median duration of 77 days (fig. 12C). FIG. 12D shows +.>As a result. In full SD patients (n=6), CR was maintained for a median duration of 57 days (fig. 12D).
FIGS. 13A-13B show overall disease improvement as demonstrated by physician global assessment (Phys-GA) and patient global assessment (Pat-GA). Phys-GA and Pat-GA were used to evaluate disease severity using the Likert scale of 0-3, where 0 is absent and 3 is severe.
Figures 14A-14D demonstrate that mAb1 treatment significantly depletes skin mast cells and serum tryptase. Fig. 14A shows mAb1 reduced skin mast cell number (n=14, p<0.05 represents p<0.01 x represents p<0.001, and p<0.0001). Fig. 14B shows mAb1 reduced serum tryptase below detection in all patients (the tryptase values below the assay limit (lloq=1 ng/mL) were normalized to 0). Figure 14C shows the kinetics of mast cells and tryptase. FIG. 14D shows that the number of skin mast cells correlates with serum tryptase levels (p<0.0001；R 2 ＝0.45)。
FIGS. 15A-15D show skin mast cell and tryptase eliminationThe kinetics of consumption reflect a decrease in the threshold of excitation. FIG. 15A shows mast cell kinetics and over time in ColdU patients As a result. FIG. 15B shows mast cell kinetics and +.>As a result. FIG. 15C shows tryptase kinetics and +.>Results (normalized to 0 for tryptase values below LLoQ; 3 ℃ for critical temperature threshold below 4 ℃ (negative test) were assigned). FIG. 15D shows tryptase kinetics and over time in SD patientsAs a result.
Figures 16A-16D show that the hematological parameters generally remained within normal ranges and a slight, transient and asymptomatic decrease in hemoglobin and White Blood Cell (WBC) parameters was noted. Fig. 16A shows hemoglobin (HgB) levels over time. Figure 16B shows WBC counts over time. Fig. 16C shows platelet counts over time. Fig. 16D shows Absolute Neutrophil Count (ANC) over time. In each figure, the hatched area indicates the corresponding normal range.
5. Detailed description of the invention
Provided herein are methods of protecting, treating, or controlling chronic prurigo, including prurigo nodularis, in a subject in need thereof comprising administering to a subject in need thereof a therapeutically effective amount of a human KIT inhibitor. The human KIT inhibitor may be any inhibitor that inhibits the activity or expression level of a human KIT protein (e.g., a human KIT protein comprising the amino acid sequence set forth in SEQ ID NO: 1). In a specific embodiment, the human KIT inhibitor is a biologic. In particular embodiments, the human KIT inhibitor is an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT. In particular embodiments, the human KIT inhibitor is a small molecule inhibitor. In specific embodiments, the human KIT inhibitor is an oligonucleotide, such as an aptamer, shRNA, miRNA, siRNA, or antisense DNA.
In one aspect, provided herein is a method of protecting, treating, or controlling chronic prurigo in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody (e.g., a KIT polypeptide comprising a human KIT D4 or D5 domain, such as a human KIT protein comprising the amino acid sequence shown in SEQ ID NO: 1) or antigen-binding fragment thereof that immunospecifically binds to human KIT.
In another aspect, provided herein is a method of protecting chronic prurigo in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody (e.g., a KIT polypeptide comprising a human KIT D4 or D5 domain, such as a human KIT protein comprising the amino acid sequence set forth in SEQ ID NO: 1) or antigen-binding fragment thereof that immunospecifically binds to human KIT.
In another aspect, provided herein are methods of treating chronic prurigo in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody (e.g., a KIT polypeptide comprising a human KIT D4 or D5 domain, such as a human KIT protein comprising the amino acid sequence set forth in SEQ ID NO: 1) or antigen-binding fragment thereof that immunospecifically binds to human KIT.
In another aspect, provided herein is a method of controlling chronic prurigo in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an antibody (e.g., a KIT polypeptide comprising a human KIT D4 or D5 domain, such as a human KIT protein comprising the amino acid sequence set forth in SEQ ID NO: 1) or antigen-binding fragment thereof that immunospecifically binds to human KIT.
In a specific embodiment, the chronic prurigo is prurigo nodularis. In a specific embodiment, the chronic prurigo is a epidemic prurigo (popular prurigo). In a specific embodiment, the chronic prurigo is prurigo nodularis. In a specific embodiment, the chronic prurigo is a plaque prurigo. In a specific embodiment, the chronic prurigo is an umbilical prurigo. In a specific embodiment, the chronic prurigo is a linear prurigo. In some embodiments, the subject exhibits a unimodal phenotype of the lesion. In other embodiments, the subject exhibits a polymorphic phenotype of the lesion.
In certain embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat, control, prevent, or protect both chronic prurigo (e.g., prurigo nodularis) and urticaria (e.g., chronic urticaria, including chronic idiopathic urticaria, and chronic induced urticaria (i.e., chronic induced urticaria (CIndU)), such as cold urticaria (cold u), symptomatic skin Scarification (SD), cholinergic urticaria, febrile urticaria, tardive pressure urticaria, solar urticaria, vibratory urticaria, contact urticaria, or waterborne urticaria). In particular embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat, control, prevent, or protect both chronic prurigo (e.g., prurigo nodularis) and chronic induced urticaria (i.e., CIndU), such as cold u, SD, hot urticaria, delayed pressure urticaria, solar urticaria, vibratory urticaria, contact urticaria, or waterborne urticaria. In particular embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat, control, prevent, or protect both chronic prurigo (e.g., prurigo nodularis) and SD. In particular embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat, control, prevent, or protect both prurigo nodularis and SD.
In various embodiments, a subject with a chronic prurigo fails one or more prior treatments for the chronic prurigo. In certain embodiments, the one or more prior treatments include at least one standard of care regimen for chronic prurigo. In certain embodiments, the one or more prior treatments are all standard-of-care therapies for chronic prurigo. In certain embodiments, the patient isSubjects with chronic prurigo are treated with an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., dupu Li Youshan anti-Is a failure to treat. In certain embodiments, a subject with chronic prurigo fails treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., nemolizumab, as described above. In certain embodiments, a subject with chronic prurigo fails treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or abrutinib. In certain embodiments, a subject with chronic prurigo fails treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., selopitane (serlopitant). In certain embodiments, a subject with chronic prurigo fails treatment with a PDE4 and/or TNF-a inhibitor, e.g., apremilast. In certain embodiments, a subject with chronic prurigo fails treatment with an OSMR β inhibitor, such as an anti-OSMR β antibody, e.g., vixarelimab. In certain embodiments, a subject with a chronic prurigo fails one, two, three, or more of the above treatments for a chronic prurigo.
If the chronic prurigo is refractory to the treatment, tolerates the treatment, relapses after the treatment and/or if the subject stops the treatment due to intolerance to the treatment, the subject is deemed to fail treatment for the chronic prurigo.
In various embodiments, the chronic prurigo is refractory to one or more prior treatments of the chronic prurigo. In certain embodiments, the one or more prior treatments include at least one standard of care regimen for the chronic prurigo. In certain embodiments, the one or more prior treatments are all standard-of-care therapies for chronic prurigo. In certain embodiments, chronic prurigo is indicated for use with an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a Dupu Li Youshan anti-antibodyIs refractory to the treatment. In certain embodiments, chronic prurigo is refractory to treatment with an inhibitor of IL-31 receptor alpha, such as an anti-IL-31 receptor alpha antibody, e.g., as ne Mo Zhushan anti (nemolizumab). In certain embodiments, the chronic prurigo is refractory to treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or ibuprotinib. In certain embodiments, chronic prurigo is refractory to treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant). In certain embodiments, the chronic prurigo is refractory to treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast. In certain embodiments, chronic prurigo is refractory to treatment with an osmrp inhibitor, such as an anti-osmrp antibody, e.g., vixarelimab. In certain embodiments, the chronic prurigo is refractory to one, two, three or more of the above treatments for the chronic prurigo.
In various embodiments, the chronic prurigo is resistant to one or more prior treatments of the chronic prurigo. In certain embodiments, the one or more prior treatments include at least one standard of care regimen for the chronic prurigo. In certain embodiments, the one or more prior treatments are all standard-of-care therapies for chronic prurigo. In certain embodiments, chronic prurigo tolerance uses an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a Dupu Li Youshan antibodyIs a therapeutic agent. In certain embodiments, chronic prurigo tolerates treatment with an inhibitor of IL-31 receptor alpha, such as an anti-IL-31 receptor alpha antibody, e.g., as a ne Mo Zhushan anti (nemolizumab). In certain embodiments, the chronic prurigo is resistant to treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or abrutinib. In certain embodiments, the chronic prurigo is resistant to treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., selopitane (serlopitant). In certain embodiments, the chronic prurigo is resistant to treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast. In some embodimentsIn chronic prurigo is resistant to treatment with osmrp inhibitors, such as anti-osmrp antibodies, e.g., vixarelimab. In certain embodiments, the chronic prurigo is resistant to one, two, three or more of the above treatments for chronic prurigo.
In various embodiments, the chronic prurigo is refractory and tolerated for one or more prior treatments of the chronic prurigo. In certain embodiments, the one or more prior treatments include at least one standard of care regimen for the chronic prurigo. In certain embodiments, the one or more prior treatments are all standard-of-care therapies for chronic prurigo. In certain embodiments, chronic prurigo is indicated for use with an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a Dupu Li Youshan anti-antibodyIs refractory and tolerogenic. In certain embodiments, chronic prurigo is refractory and resistant to treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., nemolizumab, as described in net Mo Zhushan. In certain embodiments, the chronic prurigo is refractory and resistant to treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or abrutinib. In certain embodiments, chronic prurigo is refractory and resistant to treatment with neurokinin-1 (NK 1) receptor antagonists, e.g., sellopitant (serlopitant). In certain embodiments, the chronic prurigo is refractory and resistant to treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast. In certain embodiments, chronic prurigo is refractory and resistant to treatment with osmrp inhibitors, such as anti-osmrp antibodies, e.g., vixarelimab. In certain embodiments, the chronic prurigo is refractory and tolerant to one, two, three or more of the above treatments for the chronic prurigo.
In various embodiments, the chronic prurigo recurs after one or more prior treatments of the chronic prurigo. In certain embodiments, the one or more prior treatments include at least one standard of care regimen for the chronic prurigo.In certain embodiments, the one or more prior treatments are all standard-of-care therapies for chronic prurigo. In certain embodiments, chronic prurigo is treated with an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a Dupu Li Youshan anti-IL-4R antibodyIs recurrent after treatment of (2). In certain embodiments, chronic prurigo recurs after treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., neolizumab, for example, as Ne Mo Zhushan. In certain embodiments, chronic prurigo recurs after treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or abrotinib. In certain embodiments, chronic prurigo recurs after treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant). In certain embodiments, chronic prurigo recurs after treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast. In certain embodiments, chronic prurigo recurs after treatment with an osmrp inhibitor, such as an anti-osmrp antibody, e.g., vixarelimab. In certain embodiments, the chronic prurigo recurs after one, two, three or more of the above treatments for chronic prurigo.
In various embodiments, a subject with a chronic prurigo ceases one or more prior treatments for the chronic prurigo due to intolerance of the treatment. In certain embodiments, the one or more prior treatments include at least one standard of care regimen for the chronic prurigo. In certain embodiments, the one or more prior treatments are all standard-of-care therapies for chronic prurigo. In certain embodiments, a subject with chronic prurigo ceases to use an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a Dupu Li Youshan antibody, due to treatment intoleranceIs a therapeutic agent. In certain embodiments, a subject with chronic prurigo ceases to use IL-31 receptor alpha inhibition due to intolerance of treatmentAgents, such as anti-IL-31 receptor alpha antibodies, e.g., treatment of ne Mo Zhushan anti (nemolizumab). In certain embodiments, the subject with chronic prurigo ceases treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or abrutinib, due to intolerance of the treatment. In certain embodiments, a subject with chronic prurigo ceases treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant), due to intolerance of the treatment. In certain embodiments, a subject with chronic prurigo ceases treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast, due to intolerance of the treatment. In certain embodiments, a subject with chronic prurigo ceases treatment with an osmrp inhibitor, such as an anti-osmrp antibody, e.g., vixarelimab, due to intolerance of the treatment. In certain embodiments, a subject with a chronic prurigo ceases one, two, three or more of the above treatments for a chronic prurigo due to intolerance of the treatment. / >
In various embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the subject with urticaria (e.g., chronic induced urticaria) fails to treat one or more prior treatments of urticaria. In certain embodiments, the one or more prior treatments include at least one standard of care therapy for urticaria. In certain embodiments, the one or more prior treatments are all standard-of-care therapies for urticaria. In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the subject suffering from urticaria (e.g., chronic induced urticaria) fails to treat the antihistamine of urticaria. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), Subjects with urticaria (e.g., chronically induced urticaria) fail H1-antihistamine treatment for urticaria. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), a subject suffering from urticaria (e.g., chronic induced urticaria) fails to treat H2-antihistamine for urticaria. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the subject with urticaria (e.g., chronic induced urticaria) fails to treat both H1-and H2-antihistamines of urticaria. In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), a subject suffering from urticaria (e.g., chronic induced urticaria) fails to treat the urticaria with one or more of the chlorotriene receptor antagonists. In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the subject suffering from urticaria (e.g., chronic induced urticaria) fails to treat the urticaria with one or more immunomodulatory or anti-inflammatory agents. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), a subject suffering from urticaria (e.g., chronic-induced urticaria) uses an IgE inhibitor, such as an anti-IgE antibody, e.g., omalizumab, for urticaria Or Li Geli bead mab (ligelizumab). anti-KIT antibodies, antigen-binding fragments thereof, as described hereinOr conjugates are administered to subjects to treat or control both chronic prurigo and urticaria (e.g., chronically induced urticaria), subjects suffering from urticaria (e.g., chronically induced urticaria) use IL-4R inhibitors, such as anti-IL-4R antibodies, e.g., dupu Li Youshan anti-jersey, for urticaria>Is a failure to treat. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), a subject suffering from urticaria (e.g., chronic-induced urticaria) uses an IL-5R inhibitor, such as an anti-IL-5R antibody, e.g., benralizumab (benralizumab), for urticaria>Is a failure to treat. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), a subject suffering from urticaria (e.g., chronic-induced urticaria) fails to treat urticaria with an IL-5 inhibitor, such as an anti-IL-5 antibody, e.g., meperiab. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), a subject with urticaria (e.g., chronic induced urticaria) fails to treat urticaria with a Siglec 8 inhibitor, such as an anti-Siglec 8 antibody, e.g., li Lunte power mab (lirentelimab), for urticaria. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), a subject suffering from urticaria (e.g., chronic-induced urticaria) uses a TSLP or TSLPR inhibitor, such as an anti-urticaria, for urticaria TSLP or anti-TSLPR antibodies, e.g., terlipzumab (Tezspirare) TM ) Is a failure to treat. In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), a subject suffering from urticaria (e.g., chronic induced urticaria) fails to treat urticaria with a C5aR inhibitor, such as an anti-C5 aR antibody, e.g., ai Duoli mab (avdoralimab). In particular embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), a subject suffering from urticaria (e.g., chronic-induced urticaria) fails to treat urticaria with a CD200R inhibitor, such as an anti-CD 200R antibody, e.g., LY 3454738. In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), a subject suffering from urticaria (e.g., chronic-induced urticaria) fails to treat urticaria with one or more Bruton's Tyrosine Kinase (BTK) inhibitors, e.g., lei Mibu rutinib (remibrutinib) and/or rizabetinib (rilzabrutinib). In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the following treatment of urticaria fails in a subject with urticaria (e.g., chronic induced urticaria): (1) antihistamine treatment (e.g., H1-and/or H2-antihistamine treatment), (2) treatment with one or more chlorotriene receptor antagonists, (3) treatment with one or more immunomodulators or anti-inflammatory agents (e.g., igE inhibitors, such as anti-IgE antibodies, e.g., omalizumab- >Or Li Geli bead monoclonal antibodies (ligelizumab), IL-4R inhibitors, e.g. anti-IL-4R antibodiesFor example, dupu Li Youshan is anti +.>IL-5R inhibitors, e.g. anti-IL-5R antibodies, e.g. benralizumab>IL-5 inhibitors, such as anti-IL-5 antibodies, e.g., meperiab, siglec 8 inhibitors, such as anti-Siglec 8 antibodies, e.g., li Lunte force mab (lirentelimab), TSLP or TSLPR inhibitors, such as anti-TSLP or anti-TSLPR antibodies, e.g., tertuzumab (Tezspirare) TM ) Treatment of C5aR inhibitors, such as anti-C5 aR antibodies, e.g., ai Duoli mab (avdoralimab) and/or CD200R inhibitors, such as anti-CD 200R antibodies, e.g., LY 3454738) and/or (4) treatment with one or more BTK inhibitors, e.g., lei Mibu lutinib (remibrotinib) and/or rizatrinib (rilzabrotinib). In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the following treatment of urticaria fails in a subject with urticaria (e.g., chronic induced urticaria): (1) Antihistamine treatment (e.g., H1-and/or H2-antihistamine treatment) and (2) treatment with one or more immunomodulators or anti-inflammatory agents (e.g., igE inhibitors, such as anti-IgE antibodies, e.g., omalizumab Or Li Geli bead mab (ligelizumab), IL-4R inhibitors, e.g. anti-IL-4R antibodies, e.g. Dupu Li Youshan anti +.>IL-5R inhibitors, e.g. anti-IL-5R antibodies, e.g. benralizumabIL-5 inhibitors, e.g. anti-IL-5 anti-Body, e.g., meperiab, siglec 8 inhibitors, such as anti-Siglec 8 antibodies, e.g., li Lunte force mab (lirentelimab), TSLP or TSLPR inhibitors, such as anti-TSLP or anti-TSLPR antibodies, e.g., tertuzumab (tezepelumab) (Tezspire TM ) A C5aR inhibitor, such as an anti-C5 aR antibody, e.g., ai Duoli mab (avdoralimab) and/or a CD200R inhibitor, such as an anti-CD 200R antibody, e.g., LY 3454738). In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the following treatment of urticaria fails in a subject with urticaria (e.g., chronic induced urticaria): (1) Antihistaminic therapy (e.g., H1-and/or H2-antihistamine therapy), (2) use of IgE inhibitors, such as anti-IgE antibodies, e.g., omalizumab +. >Or Li Geli bead mab (ligelizumab) and (3) use of IL-4R inhibitors, such as anti-IL-4R antibodies, e.g., dupu Li Youshan antibody->Is a therapeutic agent. In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic induced urticaria), the following treatment of urticaria fails in a subject with urticaria (e.g., chronic induced urticaria): (1) Antihistaminic therapy (e.g., H1-and/or H2-antihistaminic therapy) and (2) use of IgE inhibitors, such as anti-IgE antibodies, e.g., omalizumab +.>Or Li Geli bead mab (ligelizumab). In certain embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria),subjects with urticaria (e.g., chronically induced urticaria) fail the following treatment for urticaria: (1) Using IgE inhibitors, e.g. anti-IgE antibodies, e.g. omalizumab +>Or Li Geli bead mab (ligelizumab) and (2) use of IL-4R inhibitors, such as anti-IL-4R antibodies, e.g., dupu Li Youshan antibody- >Is a therapeutic agent.
A subject is considered to fail treatment for urticaria (e.g., chronically induced urticaria) if the urticaria (e.g., chronically induced urticaria) is refractory to the treatment, is resistant to the treatment, recurs after the treatment, and/or if the subject ceases treatment due to intolerance to the treatment. In particular embodiments, a subject having both chronic prurigo and urticaria (e.g., chronically induced urticaria) has one or more urticaria refractory to the prior treatment of urticaria (e.g., chronically induced urticaria), which may be one or more of the treatments described above for urticaria. In particular embodiments, a subject having both chronic prurigo and urticaria (e.g., chronically induced urticaria) has one or more urticaria (e.g., chronically induced urticaria) that is tolerated for one or more prior treatments of the urticaria, which may be one or more of the treatments described above for urticaria. In particular embodiments, a subject having both chronic prurigo and urticaria (e.g., chronically induced urticaria) has refractory and resistant urticaria (e.g., chronically induced urticaria) to one or more prior treatments of urticaria, which may be one or more of the treatments described above for urticaria. In particular embodiments, a subject having both chronic prurigo and urticaria (e.g., chronically induced urticaria) has urticaria (e.g., chronically induced urticaria) that recurs after one or more prior treatments of the urticaria, which may be one or more of the treatments described above for urticaria. In particular embodiments, a subject suffering from both chronic prurigo and urticaria (e.g., chronically induced urticaria) has stopped one or more prior treatments of the urticaria, which may be one or more of the treatments described above for urticaria.
In various embodiments in which an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control both chronic prurigo and urticaria (e.g., chronic-induced urticaria), the subject fails one or more prior treatments for chronic prurigo, which may be one or more treatments for chronic prurigo described above, and the subject fails one or more prior treatments for urticaria (e.g., chronic-induced urticaria), which may be one or more treatments for urticaria described above.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo, which subject has symptoms despite one or more prior treatments for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject suffering from chronic prurigo, although the use of an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti- But the subject still has symptoms of chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo, which subject is symptomatic despite treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., as a ne Mo Zhushan anti (nemolizumab). In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject suffering from chronic diseaseThe prurigo subjects, despite treatment with Janus kinase 1 inhibitors, e.g., INCB054707 or ibuproteib, still have symptoms of chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo, which subject is symptomatic despite treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., sulpirtine (serlopitant). In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo, which subject has symptoms despite treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo, which subject is symptomatic despite treatment with an OSMR β inhibitor, such as an anti-OSMR β antibody, e.g., vixarelimab.
In particular embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronic-induced urticaria), although one or more prior treatments for urticaria (which may be one or more of the treatments for urticaria described above) are performed, the subject's urticaria (e.g., chronic-induced urticaria) is still symptomatic.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronic-induced urticaria), the subject's chronic prurigo is symptomatic despite one or more prior treatments for chronic prurigo (which may be one or more treatments for chronic prurigo described above) and the subject's urticaria is symptomatic despite one or more prior treatments for urticaria (which may be one or more treatments for urticaria described above).
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following one or more prior treatments of chronic prurigo, wherein the subject fails one or more prior treatments of the chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby administering an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-for chronic prurigoWherein the subject fails the treatment with the IL-4R inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., ni Mo Lizhu mab, for chronic prurigo, wherein the subject fails the treatment with the IL-31 receptor alpha inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo, wherein the subject fails the treatment with the Janus kinase 1 inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant), for chronic prurigo, wherein the subject fails the treatment with the NK1 receptor antagonist. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby treating chronic prurigo Post-treatment with PDE4 and/or TNF- α inhibitors, e.g., apremilast, treats or controls chronic prurigo, wherein the subject fails the treatment with PDE4 and/or TNF- α inhibitors. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an OSMR inhibitor, such as an anti-OSMR antibody, for example, vixarelimab, for chronic prurigo, wherein the subject fails the treatment with the OSMR inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo after one, two, three, or more of the above treatments of chronic prurigo, wherein the subject fails the one, two, three, or more treatments.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control chronic prurigo following one or more prior treatments of chronic prurigo, wherein the chronic prurigo is refractory to the one or more prior treatments of chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby administering an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-for chronic prurigo Wherein the chronic prurigo is refractory to the treatment with the IL-4R inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, for example, as a ne Mo Zhushan anti (nemolizumab) for chronic prurigo, wherein the chronic prurigo is refractory to the treatment with the IL-31 receptor alpha inhibitor. In one embodiment, the anti-cancer agents described herein are as follows-a KIT antibody, antigen-binding fragment thereof, or conjugate thereof is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo, wherein the chronic prurigo is refractory to the treatment with the Janus kinase 1 inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopyutan (serlopitant), for chronic prurigo, wherein the chronic prurigo is refractory to the treatment with the NK1 receptor antagonist. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a PDE4 and/or TNF-a inhibitor, e.g., apremilast, for chronic prurigo, wherein the chronic prurigo is refractory to the treatment with the PDE4 and/or TNF-a inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an OSMR inhibitor, such as an anti-OSMR antibody, for example, vixarelimab, for chronic prurigo, wherein the chronic prurigo is refractory to the treatment with the OSMR inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following one, two, three, or more of the above treatments of chronic prurigo, wherein the chronic prurigo is refractory to the one, two, three, or more treatments.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following one or more prior treatments of chronic prurigo, wherein the chronic prurigo tolerates one or more prior treatments of the chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administeredIn subjects, thereby using an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a Dupu Li Youshan anti-Wherein the chronic prurigo is resistant to the treatment with the IL-4R inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, for example, as a ne Mo Zhushan anti (nemolizumab) for chronic prurigo, wherein the chronic prurigo is resistant to the treatment with the IL-31 receptor alpha inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo, wherein the chronic prurigo is resistant to the treatment with the Janus kinase 1 inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopyutan (serlopitant), for chronic prurigo, wherein the chronic prurigo is resistant to the treatment with the NK1 receptor antagonist. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a PDE4 and/or TNF-a inhibitor, e.g., apremilast, for chronic prurigo, wherein the chronic prurigo is resistant to the treatment with the PDE4 and/or TNF-a inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an OSMR inhibitor, such as an anti-OSMR antibody, for example, vixarelimab, for chronic prurigo, wherein the chronic prurigo is resistant to the treatment with an OSMR inhibitor. In one embodiment The anti-KIT antibodies, antigen-binding fragments thereof, or conjugates described herein are administered to a subject to treat or control chronic prurigo following one, two, three, or more of the above treatments of chronic prurigo, wherein the chronic prurigo tolerates the one, two, three, or more treatments.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject to treat or control chronic prurigo following one or more prior treatments of chronic prurigo, wherein the chronic prurigo is refractory and tolerated by the one or more prior treatments of chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby administering an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-for chronic prurigoWherein the chronic prurigo is refractory and tolerated for the treatment with the IL-4R inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, for example, as a ne Mo Zhushan anti (nemolizumab) for chronic prurigo, wherein the chronic prurigo is refractory and resistant to the treatment with the IL-31 receptor alpha inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo, wherein the chronic prurigo is refractory and resistant to the treatment with the Janus kinase 1 inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby reducing the risk of chronic prurigo using a neurokinin-1 (NK 1) receptor antagonist, e.g., selopium (se rlopiant) is used to treat or control chronic prurigo, wherein the chronic prurigo is refractory and resistant to the treatment with NK1 receptor antagonist. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a PDE4 and/or TNF-a inhibitor, e.g., apremilast, for chronic prurigo, wherein the chronic prurigo is refractory and resistant to the treatment with the PDE4 and/or TNF-a inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an OSMR inhibitor, such as an anti-OSMR antibody, for example, vixarelimab, for chronic prurigo, wherein the chronic prurigo is refractory and resistant to the treatment with the OSMR inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following one, two, three, or more of the above treatments of chronic prurigo, wherein the chronic prurigo is refractory and resistant to the one, two, three, or more treatments.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following one or more prior treatments of the chronic prurigo, wherein the chronic prurigo recurs following one or more prior treatments of the chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby administering an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-for chronic prurigoWherein the chronic prurigo recurs after the treatment with the IL-4R inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described hereinThe subject is administered to a subject to treat or control chronic prurigo following treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., neolizumab, for chronic prurigo, wherein the chronic prurigo recurs following the treatment with the IL-31 receptor alpha inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo, wherein the chronic prurigo recurs following the treatment with the Janus kinase 1 inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopyutan (serlopitant), for chronic prurigo, wherein the chronic prurigo recurs following the treatment with NK1 receptor antagonist. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a PDE4 and/or TNF-a inhibitor, e.g., apremilast, for chronic prurigo, wherein the chronic prurigo recurs following the treatment with the PDE4 and/or TNF-a inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an OSMR β inhibitor, such as an anti-OSMR β antibody, for example, vixarelimab, for chronic prurigo, wherein the chronic prurigo recurs following the treatment with the OSMR β inhibitor. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following one, two, three, or more of the above treatments of chronic prurigo, wherein the chronic prurigo recurs following the one, two, three, or more treatments.
In various embodiments, the anti-KIT antibodies, antigens thereof, described herein will beThe binding fragment or conjugate is administered to a subject to treat or control chronic prurigo following one or more prior treatments of chronic prurigo, wherein the subject ceases the one or more prior treatments of chronic prurigo due to intolerance of the treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby administering an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-for chronic prurigoWherein the subject ceases the treatment with the IL-4R inhibitor due to intolerance of the treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, for example, nemolizumab, for chronic prurigo, wherein the subject ceases treatment with the IL-31 receptor alpha inhibitor due to intolerance of treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo, wherein the subject has terminated the treatment with the Janus kinase 1 inhibitor due to intolerance of the treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopyutan (serlopitant), for chronic prurigo, wherein the subject terminates the treatment with the NK1 receptor antagonist due to intolerance of the treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is administered to a subject, thereby following treatment with PDE4 and/or TNF-a inhibitors for chronic prurigo, e.g., apremilast, or Controlling chronic prurigo, wherein the subject ceases the treatment with PDE4 and/or TNF-a inhibitors due to intolerance of the treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo following treatment with an OSMR inhibitor, such as an anti-OSMR antibody, e.g., vixarelimab, for chronic prurigo, wherein the subject has terminated the treatment with the OSMR inhibitor due to intolerance of the treatment. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling chronic prurigo after one, two, three, or more of the above treatments of chronic prurigo, wherein the subject terminates the one, two, three, or more treatments due to intolerance of the treatment.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling both chronic prurigo and urticaria (e.g., chronic-induced urticaria) following treatment with one or more prior treatments for urticaria, which may be one or more of the treatments described above for urticaria, wherein the subject failed one or more prior treatments for the urticaria.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject, thereby treating or controlling both chronic prurigo and urticaria (e.g., chronic-induced urticaria) after treatment with one or more prior treatments for chronic prurigo, which may be one or more of the treatments for chronic prurigo described above, wherein the subject failed the one or more prior treatments for chronic prurigo, which may be one or more of the treatments for urticaria described above, wherein the subject failed the one or more prior treatments for urticaria.
In addition, provided herein are combination therapies for the treatment of chronic prurigo comprising administering an anti-KIT antibody (e.g., humanized anti-KIT antibody) or antigen-binding fragment thereof (e.g., KIT binding fragment thereof) or antibody conjugate thereof described herein to a subject in need thereof in combination with one or more other therapies (e.g., a second therapeutic agent). In various embodiments, the combination therapy is administered to the subject substantially simultaneously, on the same day, on the same week, or on the same treatment cycle, or on similar or overlapping dosage administration schedules. In some embodiments, the combination therapy is administered to the subject simultaneously, concurrently or concomitantly. In other embodiments, the combination therapy is administered sequentially to the subject. In one embodiment, the anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to the subject prior to other therapies in the combination. In another embodiment, the anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to the subject following other therapies in the combination. The use of the term "combination" does not limit the order in which one or more anti-KIT antibodies and one or more other therapies are administered to a subject. Also provided herein are combination therapies for treating urticaria (e.g., chronically induced urticaria), as well as both combination therapies for treating chronic prurigo and combination therapies for treating urticaria (e.g., chronically induced urticaria), for subjects suffering from both chronic prurigo and urticaria (e.g., chronically induced urticaria).
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with one or more treatments for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is combined with an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-antibody, for use in chronic prurigoIs administered to a subject suffering from chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with a therapeutic combination using an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., nemolizumab, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with a treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or abbrotinib, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with a treatment using a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant), for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with a therapeutic combination using PDE4 and/or TNF-a inhibitors for chronic prurigo, e.g., apremilast. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with a therapeutic combination using an osmrp inhibitor, such as an anti-osmrp antibody, e.g., vixarelimab, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo in combination with one, two, three, or more of the above treatments for chronic prurigo.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronic-induced urticaria) in combination with one or more treatments for urticaria, which may be one or more treatments for urticaria described above.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronically induced urticaria) in combination with one or more treatments for chronic prurigo, which may be one or more treatments for chronic prurigo described above, and in combination with one or more treatments for urticaria, which may be one or more treatments for urticaria described above.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo simultaneously, concurrently, or concomitantly with one or more treatments for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is combined with an IL-4R inhibitor, such as an anti-IL-4R antibody, e.g., a dopen Li Youshan anti-antibody, for use in chronic prurigo Simultaneously, concurrently or concomitantly to a subject suffering from chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo concurrently, or concomitantly with treatment with an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., nemolizumab, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo concurrently, or concomitantly with treatment with a Janus kinase 1 inhibitor, e.g., INCB054707 or albrotinib, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo simultaneously, concurrently, or concomitantly with treatment with a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant), for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described hereinThe subject is administered to a subject suffering from chronic prurigo concurrently, concurrently or concomitantly with treatment with PDE4 and/or TNF-a inhibitors, e.g., apremilast, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo concurrently, or concomitantly with treatment with an osmrp inhibitor, such as an anti-osmrp antibody, for example, vixarelimab, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo simultaneously, concurrently, or concomitantly with one, two, three, or more of the above treatments for chronic prurigo.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronic-induced urticaria) simultaneously, concurrently, or concomitantly with one or more treatments for urticaria, which may be one or more of the treatments described above for urticaria.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronically induced urticaria) simultaneously, concurrently or concomitantly with, and with one or more treatments for chronic prurigo, which may be one or more treatments for chronic prurigo described above, and which may be one or more treatments for urticaria described above.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is sequentially administered to a subject suffering from chronic prurigo with one or more treatments for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is combined with an IL-4R inhibitor for use in chronic prurigo, e.g. anti-IL-4R antibodies, e.g., dupu Li Youshan anti-Sequentially to subjects suffering from chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is sequentially administered to a subject suffering from chronic prurigo with a treatment using an IL-31 receptor alpha inhibitor, such as an anti-IL-31 receptor alpha antibody, e.g., nemolizumab, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is sequentially administered to a subject suffering from chronic prurigo with a treatment using a Janus kinase 1 inhibitor, e.g., INCB054707 or abbrotinib, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is sequentially administered to a subject suffering from chronic prurigo with a treatment using a neurokinin-1 (NK 1) receptor antagonist, e.g., telopitane (serlopitant), for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment or conjugate thereof described herein is sequentially administered to a subject suffering from chronic prurigo with treatment with PDE4 and/or TNF-a inhibitors for chronic prurigo, e.g., apremilast. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is sequentially administered to a subject suffering from chronic prurigo with a treatment using an osmrp inhibitor, such as an anti-osmrp antibody, e.g., vixarelimab, for chronic prurigo. In one embodiment, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from chronic prurigo, in sequence with one, two, three, or more of the above treatments for chronic prurigo.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronic-induced urticaria) sequentially with one or more treatments for urticaria, which may be one or more treatments for urticaria described above.
In various embodiments, an anti-KIT antibody, antigen-binding fragment thereof, or conjugate described herein is administered to a subject suffering from both chronic prurigo and urticaria (e.g., chronically induced urticaria) sequentially with one or more treatments for chronic prurigo, which may be one or more treatments for chronic prurigo described above, and one or more treatments for urticaria, which may be one or more treatments for urticaria described above.
Also provided herein are antibodies and antigen-binding fragments thereof that immunospecifically bind to human KIT (e.g., KIT polypeptides comprising a human KIT D4 or D5 domain, such as a human KIT protein comprising the amino acid sequence set forth in SEQ ID NO: 1), which can be used in the methods described above.
As used herein, "administering" or "adminisfration" refers to the act of injecting or otherwise physically delivering a substance (e.g., a humanized anti-KIT antibody or antigen-binding fragment thereof provided herein) to a subject or patient (e.g., a human), such as by mucosal, topical, intradermal, parenteral, intravenous, intramuscular delivery, and/or any other physical delivery method described herein or known in the art.
As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of a therapy (e.g., an antibody or pharmaceutical composition provided herein) sufficient to reduce and/or ameliorate the severity and/or duration of a given disease and/or symptom associated therewith. These terms also encompass the amount necessary to reduce, slow or ameliorate the progression or development of a given disease, reduce, slow or ameliorate the recurrence, development or onset of a given disease, and/or improve or enhance the prophylactic or therapeutic effects of another therapy (e.g., a therapy other than an anti-KIT inhibitor (e.g., an anti-KIT antibody) provided herein). In some embodiments, an "effective amount" as used herein also refers to an amount of an inhibitor (e.g., an antibody) described herein that achieves the indicated result, e.g., a decrease in mast cell number and/or activity, a decrease in eosinophil number and/or activity, an inhibition (e.g., partial inhibition) of KIT biological activity of a cell, such as inhibition of cell proliferation or cell survival, or an increase or induction of apoptosis or cell differentiation, etc.
As used herein, the term "D4 or D5 region" or "D4/D5 domain" refers to the sequence from amino terminus to carboxy terminus: the D4, D4-D5 hinge region, and D5 cover regions within the KIT polypeptide of the fourth Ig-like extracellular ("D4") domain, the fifth Ig-like extracellular ("D5") domain, and the hinge region between the D4 and D5 domains ("D4-D5 hinge region"). As used herein, amino acids V308 to H515 shown in fig. 1 are considered as examples of D4/D5 regions or domains.
As used herein, the term "KIT" or "KIT receptor" or "KIT polypeptide" refers to any form of full-length KIT, including, but not limited to, native KIT, an isoform of KIT, an interspecies homolog of KIT, or a KIT variant, e.g., a naturally occurring (e.g., allelic or splice variant, or mutant, e.g., somatic mutant) or an artificially constructed variant (e.g., recombinant or chemically modified variant). KIT is a type III receptor tyrosine kinase encoded by the c-KIT gene (see, e.g., yarden et al, nature,1986,323:226-232;Ullrich and Schlessinger,Cell,1990,61:203-212; clifford et al, j. Biol. Chem.,2003,278:31461-31464; yarden et al, EMBO j.,1987,6:3341-3351; mol et al, j. Biol. Chem.,2003,278: 31461-31464). GenBank TM Accession number NM 000222 provides an exemplary human KIT nucleic acid sequence. GenBank TM Exemplary human KIT amino acid sequences are provided by accession numbers NP 001087241, PI 0721 and AAC 50969. GenBank TM Accession number AAH75716 provides an exemplary murine KIT amino acid sequence. Natural KIT comprises 5 extracellular immunoglobulin (Ig) -like domains (D1, D2, D3, D4, D5), a single transmembrane region, an inhibitory cytoplasmic membrane-proximal domain, and a cytokinin kinase domain separated by a kinase insert (see, e.g., yarden et al Nature,1986,323:226-232;Ullrich and Schlessinger,Cell,1990,61:203-212; clifford et al J.biol. Ch)em.,2003, 278:31461-31464). FIG. 1 provides exemplary amino acid sequences of the D4/D5 region of human KIT at amino acid residues V308 through H515. In a specific embodiment, KIT is human KIT. In particular embodiments, KIT may exist as monomers, dimers, multimers, natural forms, or denatured forms.
As used herein, the term "combination" in the context of administration of other therapies refers to the use of more than one therapy. The use of the term "combination" does not limit the order in which the therapies are administered. The therapies may be administered, for example, sequentially, simultaneously or concomitantly.
As used herein, the term "chronic prurigo" means a disease characterized by chronic itching (itching sensation) and the presence of multiple local or systemic prurigo lesions.
As used herein, the term "prurigo nodularis" means a disease characterized by chronic itching and the presence of multiple local or systemic, protruding, hard and nodular lesions.
As used herein, the terms "treatment" and "treatment" refer to a reduction or improvement in the development, severity, and/or duration of chronic prurigo resulting from the administration of one or more therapies, including, but not limited to, the administration of one or more prophylactic or therapeutic agents, such as antibodies provided herein.
As used herein, the terms "control" and "management" refer to the beneficial effects of a subject obtained from therapy (e.g., a prophylactic or therapeutic agent) that do not result in a cure of chronic prurigo. In certain embodiments, one or more therapies (e.g., a prophylactic or therapeutic agent, such as an antibody as described herein) are administered to a subject to "control" chronic prurigo, one or more symptoms thereof, thereby preventing the development or worsening of the disorder.
As used herein, the term "protect" or "block" in the context of chronic prurigo refers to complete or partial inhibition (e.g., less than 100%, 95%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or 5%) or blockage of chronic prurigo and/or symptoms associated therewith by administration of a therapy or combination of therapies provided herein (e.g., a prophylactic or therapeutic agent, such as a combination of antibodies as described herein).
As used herein, the term "prophylactic agent" refers to any agent that can completely or partially inhibit the progression, recurrence, onset, or spread of chronic prurigo and/or symptoms associated therewith in a subject. In certain embodiments, the term "prophylactic agent" refers to an antibody described herein. In certain other embodiments, the term "prophylactic agent" refers to agents other than the antibodies described herein. In general, a prophylactic agent is an agent known to be useful for or currently being used for preventing or impeding the onset, progression, development and/or severity of chronic prurigo and/or symptoms associated therewith. In particular embodiments, the prophylactic agent is a human anti-KIT antibody, such as a humanized or fully human anti-KIT monoclonal antibody.
As used herein, the term "side effect" or "adverse effect" encompasses both an undesirable and adverse effect of a therapy (e.g., a prophylactic or therapeutic agent). The undesired effects are not necessarily disadvantageous. Adverse effects from therapy (e.g., prophylactic or therapeutic agents) can be detrimental or uncomfortable or dangerous. Examples of side effects include diarrhea, cough, gastroenteritis, wheezing, nausea, vomiting, anorexia, abdominal cramps, fever, pain, weight loss, dehydration, hair loss, asthma, insomnia, dizziness, mucositis, nerve and muscle effects, fatigue, dry mouth and loss of appetite, rash or swelling at the site of administration, influenza-like symptoms such as fever, chills and fatigue, digestive tract problems and allergic reactions. Other undesirable effects experienced by patients are numerous and known in the art. Various effects are described in Physician's Desk Reference (71 st edition, 2017).
As used herein, the terms "subject" and "patient" are used interchangeably. As used herein, a subject is a mammal, such as a non-primate (e.g., cow, pig, horse, cat, dog, goat, rabbit, rat, mouse, etc.) or primate (e.g., monkey and human), e.g., human. In one embodiment, the subject is a mammal, e.g., a human, diagnosed with chronic prurigo. In another embodiment, the subject is a mammal, e.g., a human, at risk of developing chronic prurigo. In another embodiment, the subject is a non-human primate. In a specific embodiment, the subject is an adult. In a specific embodiment, the subject is an adult subject at least 18 years old. In particular embodiments, the subject is a child. In a specific embodiment, the subject is a child between 1 year and 18 years of age. In a specific embodiment, the subject is a human between 1 and 3 years of age. In particular embodiments, the subject is a human between 3 and 12 years of age or between 12 and 18 years of age.
As used herein, the term "therapy" may refer to any procedure, method, composition, formulation, and/or agent that may be used to prevent, protect, treat, control, or ameliorate a condition or disorder or symptom thereof, or one or more symptoms or conditions associated therewith. In certain embodiments, the term "therapy" refers to a drug therapy, adjuvant therapy, radiation, surgery, biological therapy, supportive therapy, and/or other therapies useful for protecting, treating, controlling, preventing, or ameliorating a condition or disorder or one or more symptoms thereof or one or more symptoms or conditions associated therewith. In certain embodiments, the term "therapy" refers to a therapy other than an anti-KIT antibody or pharmaceutical composition thereof described herein. In particular embodiments, "other therapies" and "additional therapies" refer to therapies other than treatment with an anti-KIT antibody or pharmaceutical composition thereof described herein. In particular embodiments, the therapy comprises the use of an anti-KIT antibody described herein as an adjuvant therapy. For example, the anti-KIT antibodies described herein are used in conjunction with drug therapies, biological therapies, surgery, and/or supportive therapies.
As used herein, the term "therapeutic agent" refers to any agent that can be used to protect, treat, control, or ameliorate chronic prurigo and/or symptoms associated therewith. In certain embodiments, the term "therapeutic agent" refers to an anti-KIT antibody or antigen-binding fragment thereof described herein. In certain other embodiments, the term "therapeutic agent" refers to an agent other than an antibody described herein. In particular embodiments, the therapeutic agent is an agent known to be useful for, or has been or is currently being used for, the protection, treatment, control or amelioration of chronic prurigo or one or more symptoms associated therewith.
As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. The terms "a" and "an" and "one or more" and "at least one" are used interchangeably herein.
It will be understood that wherever aspects are described herein by the language "comprising," similar aspects described by the terms "consisting of … …" and/or "consisting essentially of … …" are additionally provided.
As used herein and unless otherwise indicated, the terms "about" and "approximately" should be understood to allow standard changes, such as, for example, changes within 20% or 10% or 5%, at the discretion of the person skilled in the art. In particular embodiments, the terms "about" and "approximately" encompass the exact values recited.
5.1 antibodies
Provided herein are antibodies (e.g., anti-KIT antibodies) or antigen-binding fragments thereof that specifically bind to a KIT receptor (e.g., an extracellular domain of a human KIT receptor, e.g., as set forth in SEQ ID NO:1 or fig. 1) for use in a method of preventing, treating, or managing chronic prurigo.
As used herein, the terms "antibody" and "immunoglobulin" and "Ig" are terms of art and are used interchangeably herein and refer to a molecule having an antigen binding site that immunospecifically binds an antigen.
Antibodies include, for example, monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, synthetic antibodies, tetrameric antibodies comprising 2 heavy chains and 2 light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain-antibody heavy chain pairs, intracellular antibodies, heteroconjugate antibodies, single domain antibodies, monovalent antibodies, single chain antibodies, or single-chain Fv (scFv), caramelized antibodies, affibodies (afybody), fab fragments, F (ab') fragments, disulfide-linked Fv (sdFv), anti-idiotypic (anti-Id) antibodies (including, for example, anti-Id antibodies), and epitope-binding fragments of any of the foregoing antibodies. In certain embodiments, the antibodies described herein represent a polyclonal antibody population. Antibodies may be immunoglobulin molecules of any type (e.g., igG, igE, igM, igD, igA or IgY), of any class (e.g., igG1, igG2, igG3, igG4, igA1 or IgA 2), or of any subclass (e.g., igG2a or IgG2 b). In certain embodiments, the antibodies described herein are IgG antibodies or classes (e.g., human IgG1 or IgG 4) or subclasses thereof.
As used herein, an "antigen" is a moiety or molecule that contains an epitope and as such is also specifically bound by an antibody. In particular embodiments, the antigen to which an antibody described herein binds is KIT (e.g., human KIT) or a fragment thereof, e.g., the extracellular domain of KIT (e.g., human KIT) or the D4 region of KIT (e.g., human KIT).
As used herein, the terms "antigen binding domain," "antigen binding region," "antigen binding fragment," and similar terms refer to portions of an antibody molecule (e.g., complementarity Determining Regions (CDRs)) that comprise amino acid residues that interact with an antigen and confer specificity to the antigen on the antibody molecule. The antigen binding region may be derived from any animal species, such as rodents (e.g., mice, rats, or hamsters) and humans. The CDRs of an antibody molecule can be determined by any method known to those skilled in the art. Specifically, CDRs can be determined according to the Kabat numbering system (see Kabat et al (1991) Sequences of Proteins of Immunological interest. (u.s.device of Health and Human Services, washington, d.c.), 5 th edition). In certain aspects, it may be according to (i) a Chothia numbering scheme, which will be referred to herein as a "Chothia CDR" (see, e.g., chothia and Lesk,1987, J.mol. Biol,196:901-917; al-Lazikani et al, 1997, J.mol. Biol,273:927-948; and U.S. Pat. No.7,709,226); (ii) IMGT numbering systems, e.g., as described in Lefranc, m. -p.,1999,The Immunologist,7:132-136 and Lefranc, m. -p., et al, 1999,Nucleic Acids Res, 27:209-212; (iii) ABM numbering systems, for example, as described in MacCallum et al, 1996, J.mol. Biol.,262:732-745 and Martin, A., "Protein Sequence and Structure Analysis of Antibody Variable Domains," in Antibody Engineering, kontermann and Dubel Main, chapter 31, pages 422-439, springer-Verlag, berlin (2001); or (iv) a Contact numbering system that determines the CDRs of antibodies based on an analysis of available complex crystal structures (bioif. Org. Uk/abs) (see, e.g., macCallum et al, (1996) J Mol Biol 5:732-745).
As used herein, the term "constant region" or "constant domain" refers to a portion of an antibody that does not directly participate in binding of the antibody to an antigen, but that exhibits or contributes to a variety of effector functions, such as interactions with Fc receptors, e.g., the carboxy-terminal portion of the light and/or heavy chain. These terms refer to a portion of an immunoglobulin molecule having a generally more conserved amino acid sequence relative to an immunoglobulin variable domain.
As used herein, an "epitope" is a term in the art and refers to a localized region of an antigen to which an antibody can specifically bind. The region or polypeptide contributing to the epitope may be contiguous amino acids of the polypeptide or the epitope may be clustered together from two or more non-contiguous regions of the polypeptide.
When used in reference to antibodies, the term "heavy chain" as used herein refers to any of the different types of amino acid sequences based on a constant domain, e.g., α (α), δ (δ), ε (ε), γ (γ), and μ (μ), which produce antibodies of IgA, igD, igE, igG and IgM classes, including IgG subclasses, e.g. IgG 1 、IgG 2 、IgG 3 And IgG 4 . In a specific embodiment, the heavy chain is a human heavy chain.
As used herein, the terms "immunospecific binding," "immunospecific recognition," "specific binding," and "specific recognition" are similar terms in the context of antibodies and refer to molecules that bind to an antigen (e.g., an epitope or immune complex), as such binding is understood by those skilled in the art. For example, molecules that specifically bind to an antigen may bind to other peptides or polypeptides, typically with lower affinity, such as by, for example, immunoassays, biacore TM Determined by a KinExA 3000 instrument (Sapidyne Instruments, boise, ID) or other assay known in the art. In a specific embodiment, a molecule that immunospecifically binds to an antigen is used to bind K to another antigen than when the molecule is bound to another antigen a K is at least 2log, 2.5log, 3log, 4log or more greater a Binds to the antigen. In another specific embodiment, molecules that immunospecifically bind to an antigen do not cross-react with other proteins. In another specific embodiment, molecules that immunospecifically bind to an antigen do not cross-react with other non-KIT proteins.
As used herein, an "isolated" or "purified" antibody is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the antibody was obtained, or substantially free of chemical precursors or other chemicals when chemically synthesized. In specific embodiments, the antibodies or antigen binding fragments described herein are isolated.
The term "Kabat numbering" and similar terms are well-known in the art and refer to the numbering system of amino acid residues in the heavy and light chain variable regions of antibodies or antigen-binding portions thereof (Kabat et al (1971) Ann.NY Acad.Sci.190:382-391 and Kabat et al (1991) Sequences of Proteins of Immunological Interest, 5 th edition, U.S. Pat. No. of Health and Human Services, NIH Publication No. 91-3242). CDRs within an antibody heavy chain molecule are typically found at amino acid positions 31 to 35 ("CDR 1"), amino acid positions 50 to 65 ("CDR 2") and amino acid positions 95 to 102 ("CDR 3") using the Kabat numbering system. CDRs within an antibody light chain molecule are typically present at amino acid positions 24 to 34 (CDR 1), amino acid positions 50 to 56 (CDR 2) and amino acid positions 89 to 97 (CDR 3) using the Kabat numbering system.
When used in reference to antibodies, the term "light chain" as used herein refers to any of a variety of types, e.g., kappa (kappa) and lambda (lambda) based on the amino acid sequence of a constant domain. Light chain amino acid sequences are well known in the art. In a specific embodiment, the light chain is a human light chain.
As used herein, the term "monoclonal antibody" refers to an antibody obtained from a homogeneous or substantially homogeneous population of antibodies, and each monoclonal antibody will typically recognize a single epitope on the antigen. The term "monoclonal" is not limited to any particular method for producing antibodies. In general, a population of monoclonal antibodies can be produced by a cell, cell population, or cell line. In particular embodiments, as used herein, a "monoclonal antibody" is an antibody produced by a single hybridoma or other cell (e.g., a host cell that produces a recombinant antibody), wherein the antibody immunospecifically binds to a KIT epitope (e.g., an epitope of D4 of human KIT), as determined, for example, by ELISA or other antigen-binding or competitive binding assays known in the art or in the examples provided herein. For example, monoclonal antibodies described herein can be prepared by hybridoma methods, such as Kohler et al; nature,256:495 (1975), or monoclonal antibodies described herein may be isolated from phage libraries, for example, using techniques as described herein. Other methods for preparing clonal cell lines and monoclonal antibodies expressed thereby are well known in the art (see, e.g., chapter 11: short Protocols in Molecular Biology, (2002), 5 th edition, ausubel et al (major code), john Wiley and Sons, new York). In a specific embodiment, the monoclonal antibody is a monospecific antibody in which the antigen binding regions thereof are specific for the same epitope. In other specific embodiments, the monoclonal antibodies can be monovalent (having one antigen binding region) or multivalent (having more than one antigen binding region), e.g., bivalent (having 2 antigen binding regions).
As used herein, the term "naked antibody" refers to an antibody that is not linked, fused or conjugated to another agent or molecule (e.g., a label or drug), peptide, or polypeptide. In particular embodiments, the naked antibody expressed by the mammalian host cell may be glycosylated by the glycosylation machinery of the host cell, e.g., a glycosylase. In certain embodiments, the naked antibody is not glycosylated when expressed by a host cell that does not have its own glycosylation machinery, e.g., a glycosylase. In certain embodiments, the naked antibody is an intact antibody, and in other embodiments, the naked antibody is an antigen binding fragment of an intact antibody, such as a Fab antibody.
As used herein, the term "polyclonal antibody" refers to a population of antibodies that includes a plurality of different antibodies against the same or different epitopes within one or more antigens. Methods for producing polyclonal antibodies are known in the art (see, e.g., chapter 11: short Protocols in Molecular Biology, (2002), 5 th edition, ausubel et al (Main plaited), john Wiley and Sons, new York).
As used herein, the term "recombinant human antibody" includes human antibodies isolated, prepared, expressed or produced by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells, antibodies isolated from recombinant, combinatorial human antibody libraries, antibodies isolated from animals (e.g., mice, rabbits, goats or cattle) transgenic and/or transchromosomal for human immunoglobulin genes (see, e.g., taylor, l.d. et al (1992) nucleic acids res.20:6287-6295), or antibodies prepared, expressed, produced or isolated by any other means, including, for example, by: synthesis, genetic engineering of DNA sequences encoding human immunoglobulin sequences, or sequences encoding human immunoglobulins, e.g., splicing of human immunoglobulin gene sequences to other such sequences. These recombinant human antibodies may have variable and constant regions derived from human germline immunoglobulin sequences. In certain embodiments, the amino acid sequences of these recombinant human antibodies have been modified, and as such the amino acid sequences of the VH and/or VL regions of the recombinant antibodies are sequences that, although derived from and related to human germline VH and VL sequences, are not naturally occurring within the human antibody germline repertoire in vivo. As a non-limiting example, a recombinant human antibody may be obtained by assembling some human sequence fragments into a composite human sequence of the recombinant human antibody.
As used herein, the term "variable region" or "variable domain" refers to an antibody portion, typically a light chain or heavy chain portion, typically from 110 to 120 amino acids at the amino-terminus in the mature heavy chain and from about 90 to 100 amino acids in the mature light chain, which varies widely in sequence among antibodies, and is used in the binding and specificity of a particular antibody for its particular antigen. The variability in the sequences is concentrated in those regions called Complementarity Determining Regions (CDRs), while the regions that are more highly conserved in the variable domains are called Framework Regions (FR).
Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with the antigen. In specific embodiments, numbering of the amino acid positions of antibodies described herein is according to the EU index, as described in Kabat et al (1991) Sequences of Proteins of Immunological Interest, 5 th edition, U.S. Pat. No. of Health and Human Services, NIH Publication No.91-3242 ("Kabat et al"). In certain aspects, it may be according to (i) a Chothia numbering scheme, which will be referred to herein as a "Chothia CDR" (see, e.g., chothia and Lesk,1987, J.mol. Biol,196:901-917; al-Lazikani et al, 1997, J.mol. Biol,273:927-948; and U.S. Pat. No.7,709,226); (ii) IMGT numbering systems, e.g., as described in Lefranc, m. -p.,1999,The Immunologist,7:132-136 and Lefranc, m. -p., et al, 1999,Nucleic Acids Res, 27:209-212; (iii) ABM numbering systems, for example, as described in MacCallum et al, 1996, J.mol.biol.,262:732-745 and Martin, A., "Protein Sequence and Structure Analysis of Antibody Variable Domains," in Antibody Engineering, kontermann and Dubel, chapter 31, pages 422-439, springer-Verlag, berlin (2001); or (iv) a Contact numbering system that determines the CDRs of antibodies based on an analysis of available complex crystal structures (bioif. Org. Uk/abs) (see, e.g., macCallum et al, (1996) J Mol Biol 5:732-745). In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and a human Framework Region (FR). In particular embodiments, the variable region is a primate (e.g., non-human primate) variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) Framework Regions (FR). As a non-limiting example, the variable regions described herein result from assembling two or more fragments of a human sequence into a composite human sequence.
anti-KIT antibodies suitable for use in the methods provided herein may be selected as described herein.
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises: a light chain variable region ("VL") comprising VL CDRs 1-3 as shown in table 1 and a heavy chain variable region ("VH") comprising VH CDRs 1-3 as shown in table 1. In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises: a light chain variable region ("VL") comprising VL CDRs 1-3 as set forth in table 2 (group 1 or group 2) and a heavy chain variable region ("VH") comprising VH CDRs 1-3 as set forth in table 2 (group 1 or group 2). In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises: a light chain variable region ("VL") comprising VL CDRs 1-3 as shown in table 3 (AbM CDRs or Contact CDRs) and a heavy chain variable region ("VH") comprising VH CDRs 1-3 as shown in table 3 (AbM CDRs or Contact CDRs).
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises: VL comprising VL CDR 1-3 (SEQ ID NOS: 2-4) as shown in Table 1 and VH comprising VH CDR 1-3 (SEQ ID NOS: 5-7) as shown in Table 1. In particular embodiments, such anti-KIT antibodies are naked antibodies. In particular embodiments, such anti-KIT antibodies are bivalent monospecific antibodies. In a specific embodiment, such an anti-KIT antibody is a bispecific antibody. In certain embodiments, such anti-KIT antibodies are not bispecific antibodies.
Table 1: CDR amino acid sequences
Amino acid sequence SEQ ID NO：
VL CDR1 KASQNVRTNVA 2
VL CDR2 SASYRYS 3
Amino acid sequence SEQ ID NO：
VL CDR3 QQYNSYPRT 4
VH CDR1 DYYIN 5
VH CDR2 RIYPGSGNTYYNEKFKG 6
VH CDR3 GVYYFDY 7
Table 2: CDR amino acid sequences
Table 3: CDR amino acid sequences
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises:
(i) VL comprising the amino acid sequence:
DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSASYRYSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVEIK (SEQ ID NO: 17), wherein X K1 To X K6 Is any amino acid; and
(ii) VH comprising the amino acid sequence:
QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIARIYPGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGVYYFDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 To X H8 Is any amino acid.
In a specific embodiment, xκ 1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, xκ 2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, xκ 3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Is an amino acid having a charged or acidic side chain, X K6 Is an amino acid having an aromatic side chain, X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain.
In a specific embodiment, X K1 Is amino acid F or S, xkappa 2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T, X K6 Is amino acid F or Y, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises:
(i) VL comprising the amino acid sequence:
DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSASYRYSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGG
GTKVEIK (SEQ ID NO: 17), wherein X K1 To X K6 Is any amino acid; and
(ii) VH comprising the amino acid sequence comprising SEQ ID NO: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequences shown.
In a specific embodiment, xκ 1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, xκ 2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, xκ 3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Is an amino acid having a charged or acidic side chain, and X K6 Is an amino acid having an aromatic side chain.
In a specific embodiment, X K1 Is amino acid F or S, xkappa 2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T and X K6 Is amino acid F or Y.
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises:
(i) VL comprising a sequence having SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising the amino acid sequence:
QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIA RIYPGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARG VYYFDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 To X H8 Is any amino acid.
In a specific embodiment, X H1 Is an amino acid having an aliphatic side chain, X H2 Is provided with aliphatic side chainsAmino acids, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain.
In a specific embodiment, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises: a heavy chain variable region ("VH") comprising an amino acid sequence selected from Table 4 (SEQ ID NOS: 8-12) and/or a light chain variable region ("VL") comprising an amino acid sequence selected from Table 5 (SEQ ID NOS: 13-16). In particular embodiments, such anti-KIT antibodies are naked antibodies. In particular embodiments, such anti-KIT antibodies are bivalent monospecific antibodies. In a specific embodiment, such an anti-KIT antibody is a bispecific antibody. In certain embodiments, such anti-KIT antibodies are not bispecific antibodies.
Table 4: VH amino acid sequence
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Table 5: VL amino acid sequence
In a particular aspect, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises VH comprising SEQ ID NO:8, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:13, and a nucleotide sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:8, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:8, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:15, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:8, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.
In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:9, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:13, and a nucleotide sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:9, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:9, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:15, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:9, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.
In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:10, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:13, and a nucleotide sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:10, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:10, and VL comprising the amino acid sequence set forth in SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:10, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:15, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:10, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.
In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:11, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:13, and a nucleotide sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:11, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:11, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:15, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:11, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.
In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:12, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:13, and a nucleotide sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:12, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:14, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:12, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:15, and a polypeptide having the amino acid sequence shown in seq id no. In one embodiment, the anti-KIT antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO:12, and/or VL comprising the amino acid sequence set forth in SEQ ID NO:16, and a polypeptide having the amino acid sequence shown in seq id no.
In particular aspects, an anti-KIT antibody (e.g., a humanized antibody) or antigen-binding fragment thereof for use in a method of preventing, treating, or controlling chronic prurigo comprises:
(i) VL comprising the amino acid sequence: and SEQ ID NO:13 has at least 90% identity to SEQ ID NO:14 has at least 88% identity to SEQ ID NO:15, or at least 87% identical to SEQ ID NO:16 having an amino acid sequence with at least 84% identity; and
(ii) VH comprising the amino acid sequence: and SEQ ID NO:8 has at least 93% identity to SEQ ID NO:9 has at least 92% identity to SEQ ID NO:10 has at least 90% identity to SEQ ID NO:11 or at least 87% identical to SEQ ID NO:12, an amino acid sequence having at least 86% identity.
Previous anti-KIT antibodies have been found to cause degranulation of FcgRI expressing human mast cells and/or exhibit Fc receptor-dependent KIT agonist activity, which can lead to undesired infusion-related reactions (IRR) as well as other adverse effects.
In various embodiments, an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprises a modified (e.g., mutated) Fc region or domain (e.g., a modified (e.g., mutated) human IgG Fc region or domain, such as a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 Fc region or domain). Preferably, the anti-KIT antibody or antigen-binding fragment used in the methods described herein has reduced Fc receptor binding activity (specifically reduced fcγr binding activity), does not cause degranulation of FcgRI-expressing human mast cells and/or exhibits Fc receptor-dependent KIT agonist activity. In certain embodiments, one or more of these properties of the anti-KIT antibody or antigen-binding fragment is caused by a modified (e.g., mutated) Fc region or domain.
In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein have reduced Fc receptor binding activity (specifically reduced fcγr binding activity). In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein do not have significant Fc receptor (particularly fcγr) binding activity. In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein have no detectable Fc receptor (specifically fcγr) binding activity. In particular embodiments, the anti-KIT antibody or antigen-binding fragment used in the methods described herein has at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% lower Fc receptor (particularly fcγr) binding activity than an appropriate control antibody or antigen-binding fragment. When an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprises a modified (e.g., mutated) Fc region or domain (e.g., a modified (e.g., mutated) human IgG Fc region or domain, such as a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 Fc region or domain), in a preferred embodiment, the appropriate control antibody or antigen-binding fragment is an antibody or antigen-binding fragment having the same VH and VL, but the same isotype, wild-type (unmodified) Fc region or domain. In particular embodiments, an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprises a modified (e.g., mutated) human IgG1 Fc region or domain and has at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% lower Fc receptor (particularly fcγr) binding activity as compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain.
In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein do not cause significant degranulation of human mast cells expressing FcgRI (e.g., as determined, for example, by the percentage of β -hexosaminidase released from human mast cells in culture (e.g., in the presence of ifnγ). In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein do not cause detectable degranulation of human mast cells expressing FcgRI (e.g., as determined, for example, by the percentage of β -hexosaminidase released from human mast cells in culture (e.g., in the presence of ifnγ). In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein result in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% lower degranulation of human mast cells expressing FcgRI (e.g., as determined, for example, by the percentage of β -hexosaminidase released from human mast cells in culture (e.g., in the presence of ifnγ) compared to an appropriate control antibody or antigen-binding fragment. In specific embodiments, the release of β -hexosaminidase in human mast cells in culture is reduced by more than 50% in the presence of ifnγ by an anti-KIT antibody or antigen-binding fragment used in the methods described herein, as compared to an appropriate control antibody or antigen-binding fragment. In specific embodiments, the release of β -hexosaminidase in human mast cells in culture is reduced by more than 60%, more than 70%, or more than 80% in the presence of ifnγ by an anti-KIT antibody or antigen-binding fragment used in the methods described herein, as compared to an appropriate control antibody or antigen-binding fragment. When an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprises a modified (e.g., mutated) Fc region or domain (e.g., a modified (e.g., mutated) human IgG Fc region or domain, such as a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 Fc region or domain), in a preferred embodiment, the appropriate control antibody or antigen-binding fragment is an antibody or antigen-binding fragment having the same VH and VL, but the same isotype, wild-type (unmodified) Fc region or domain. In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein comprise a modified (e.g., mutated) human IgG1 Fc region or domain and result in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% lower degranulation of human mast cells expressing FcgRI (e.g., as determined, for example, by the percentage of β -hexosaminidase released from human mast cells in culture (e.g., in the presence of ifnγ) as compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain. In particular embodiments, the release of β -hexosaminidase from human mast cells in culture is reduced by more than 50% in the presence of ifny by an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprising a modified (e.g., mutated) human IgG1 Fc region or domain, as compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain. In particular embodiments, the release of β -hexosaminidase from human mast cells in culture is reduced by more than 60%, more than 70%, or more than 80% in the presence of ifny by an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprising a modified (e.g., mutated) human IgG1 Fc region or domain, as compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain.
In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein do not exhibit significant Fc receptor-dependent KIT agonistic activity (e.g., as determined, for example, by KIT phosphorylation). In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein do not exhibit detectable Fc receptor-dependent KIT agonistic activity (e.g., as determined, for example, by KIT phosphorylation). In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein elicit at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% lower Fc receptor-dependent KIT activity (e.g., as determined, for example, by KIT phosphorylation) than an appropriate control antibody or antigen-binding fragment. In particular embodiments, the Fc receptor-dependent KIT agonist activity (as determined by KIT phosphorylation using a cross-linked Fc receptor) is reduced by more than 50% using an anti-KIT antibody or antigen-binding fragment used in the methods described herein as compared to an appropriate control antibody or antigen-binding fragment. In particular embodiments, the Fc receptor-dependent KIT agonist activity (as determined by KIT phosphorylation using a cross-linked Fc receptor) is reduced by more than 60%, more than 70%, or more than 80% using an anti-KIT antibody or antigen-binding fragment used in the methods described herein, as compared to an appropriate control antibody or antigen-binding fragment. When an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprises a modified (e.g., mutated) Fc region or domain (e.g., a modified (e.g., mutated) human IgG Fc region or domain, such as a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4Fc region or domain), in a preferred embodiment, the appropriate control antibody or antigen-binding fragment is an antibody or antigen-binding fragment having the same VH and VL, but the same isotype, wild-type (unmodified) Fc region or domain. In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein comprise a modified (e.g., mutated) human IgG1 Fc region or domain and result in at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% lower Fc receptor-dependent KIT activity (e.g., as determined, for example, by KIT phosphorylation) when compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain. In particular embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein do not exhibit significant or detectable Fc receptor-dependent KIT agonistic activity as described herein, even when crosslinked on THP-1 cells. In particular embodiments, the Fc receptor-dependent KIT agonist activity (as determined by KIT phosphorylation using a cross-linked Fc receptor) is reduced by more than 50% when using an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprising a modified (e.g., mutated) human IgG1 Fc region or domain when compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain. In particular embodiments, the Fc receptor-dependent KIT agonist activity (as determined by KIT phosphorylation using a cross-linked Fc receptor) is reduced by more than 60%, more than 70%, or more than 80% using an anti-KIT antibody or antigen-binding fragment used in the methods described herein comprising a modified (e.g., mutated) human IgG1 Fc region or domain when compared to a corresponding antibody or antigen-binding fragment having the same VH and VL but having a wild-type (unmodified) human IgG1 Fc region or domain.
In various embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein (1) reduce disease activity in chronic prurigo patients, (2) reduce the number of skin mast cells in chronic prurigo patients, (3) reduce the level of tryptase in chronic prurigo patients, and/or (4) maintain hematologic parameters such as hemoglobin (HgB) levels, white Blood Cell (WBC) counts, platelet counts, and/or Absolute Neutrophil Counts (ANC) in patients, such as chronic prurigo patients, within normal ranges.
In certain embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein can significantly reduce the number of skin mast cells in a chronic prurigo patient relative to the number prior to treatment. In particular embodiments, an anti-KIT antibody or antigen-binding fragment used in the methods described herein can reduce the number of skin mast cells in a chronic prurigo patient by at least 20%, at least 40%, at least 60%, or at least 80% (e.g., within 1 week, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, or 12 weeks after treatment with the anti-KIT antibody or antigen-binding fragment) relative to the number prior to treatment. In specific embodiments, the effect of an anti-KIT antibody or antigen-binding fragment used in the methods described herein lasts for at least 2 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks.
In certain embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein can significantly reduce serum tryptase in chronic prurigo patients relative to pre-treatment levels. In particular embodiments, an anti-KIT antibody or antigen-binding fragment used in the methods described herein can reduce serum tryptase in a chronic prurigo patient by at least 50%, at least 70%, or at least 90% (e.g., within 1 week, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, or 12 weeks after treatment with the anti-KIT antibody or antigen-binding fragment) relative to a pre-treatment level. In specific embodiments, the effect of an anti-KIT antibody or antigen-binding fragment used in the methods described herein lasts for at least 2 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks.
In certain embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein maintain hematological parameters in a patient, such as hemoglobin (HgB) levels, white Blood Cell (WBC) counts, platelet counts, and/or Absolute Neutrophil Counts (ANC), within normal ranges. In certain embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein maintain hematological parameters, such as hemoglobin (HgB) levels, white Blood Cell (WBC) counts, platelet counts, and/or Absolute Neutrophil Counts (ANC), in chronic prurigo patients within normal ranges. In specific embodiments, the hematology parameter is maintained for at least 2 weeks, at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, or at least 12 weeks.
In various embodiments, the anti-KIT antibodies or antigen-binding fragments used in the methods described herein have one or more of the properties described herein.
In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue.
In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of human IgG1 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue selected from the group consisting of: 234A, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235F, 236E, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247V, 247G, 252Y, 254T, 26e, 2621, 262A, 262T, 262E, 2631, 263A, 263T, 263M, 264L, 2641, 26w, T, R, 264F, 264M, 264E, 264G, 265V, 265F, 265H, 51H, 265G, 265V, 265H, 51H numbered by the EU index as described in Kabat. 2661, 266A, 266T, 266M, 267Q, 267L, 269H, 269Y, 269F, 269R, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 269G, 297S, 297D, 297E, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 313F, 322Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 327G, 327W, 327N, 327L, 328S, 328M, 328D, 328E, 328N, 328Q, 328F, 3281, 328V, 328T, 328H, 328A, 328F, 329H, 329Q, 330K, 330G, 330T, 330C, 330L, 330Y, 330V, 3301, 330F, 330R, 330H, 332D, 332F, 332T, 332F, 332T, 332A, 332T, 332H, 332T and 332H. Optionally, the Fc region or domain may comprise additional and/or alternative non-naturally occurring amino acid residues known to those skilled in the art (see, e.g., U.S. Pat. Nos. 5,624,821;6,277,375;6,737,056; PCT patent publication WO 01/58957; WO 04/016750; WO 04/029207; WO 04/035752 and WO 05/040217). In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of a human IgG2 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue that is equivalent to an amino acid residue described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art. In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of human IgG3 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue that is equivalent to an amino acid residue described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art. In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of human IgG4 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue that is equivalent to an amino acid residue described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art.
In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of human IgG1 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue selected from the group consisting of: 234A, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235F, 236E, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247V, 247G, 252Y, 254T, 26e, 2621, 262A, 262T, 262E, 2631, 263A, 263T, 263M, 264L, 2641, 26w, T, R, 264F, 264M, 264E, 264G, 265V, 265F, 265H, 51H, 265G, 265V, 265H, 51H numbered by the EU index as described in Kabat. 2661, 266A, 266T, 266M, 267Q, 267L, 269H, 269Y, 269F, 269R, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 269G, 297S, 297D, 297E, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 313F, 322Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 327G, 327W, 327N, 327L, 328S, 328M, 328D, 328E, 328N, 328Q, 328F, 3281, 328V, 328T, 328H, 328A, 328F, 329H, 329Q, 330K, 330G, 330T, 330C, 330L, 330Y, 330V, 3301, 330F, 330R, 330H, 332D, 332F, 332T, 332F, 332T, 332A, 332T, 332H, 332T and 332H. Optionally, the Fc region or domain may comprise additional and/or alternative non-naturally occurring amino acid residues known to those skilled in the art (see, e.g., U.S. Pat. Nos. 5,624,821;6,277,375;6,737,056; PCT patent publication WO 01/58957; WO 04/016750; WO 04/029207; WO 04/035752 and WO 05/040217). In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of a human IgG2 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue that is equivalent to an amino acid residue described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art. In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of human IgG3 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue that is equivalent to an amino acid residue described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art. In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) Fc region or domain, wherein the Fc region or domain is that of human IgG4 and comprises at least one (e.g., 1, 2, 3, 4, 5, or 6) amino acid modification (e.g., substitution, deletion, or addition) or at least one (e.g., 1, 2, 3, 4, 5, or 6) non-naturally occurring amino acid residue that is equivalent to an amino acid residue described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art.
In a certain aspect, provided herein is an antibody comprising an Fc region or domain, wherein said Fc region or domain is an Fc region or domain of human IgG1 and comprises at least a non-naturally occurring amino acid at one or more positions selected from 239, 330 and 332, as numbered by the EU index as set forth in Kabat. In a specific embodiment, provided herein is an antibody comprising an Fc region or domain, wherein said Fc region or domain is an Fc region or domain of human IgG1 and comprises at least one non-naturally occurring amino acid selected from 239D, 330L, and 332E, as numbered by the EU index as set forth in Kabat. Optionally, the Fc region or domain may also include other non-naturally occurring amino acids at one or more positions selected from 252, 254, and 256, as numbered by the EU index as set forth in Kabat. In a specific embodiment, provided herein is an antibody comprising an Fc region or domain, wherein the Fc region or domain is an Fc region or domain of human IgG1 and comprises at least one non-naturally occurring amino acid selected from 239D, 330L and 332E, as numbered by the EU index as set forth in Kabat, and the at least one non-naturally occurring amino acid at one or more positions is selected from 252Y, 254T and 256E, as numbered by the EU index as set forth in Kabat. In particular embodiments, provided herein are antibodies comprising an Fc region or domain, wherein the Fc region or domain is an Fc region or domain of a human IgG2, igG3, or IgG4, and comprises at least one non-naturally occurring amino acid residue equivalent to the amino acid residues described herein for the human IgG1 Fc region or domain, as can be determined by one of skill in the art. In particular embodiments, provided herein are antibodies comprising an Fc region or domain, wherein the Fc region or domain is an Fc region or domain of human IgG2, igG3, or IgG4, and comprises at least one non-naturally occurring amino acid residue at one or more positions equivalent to those described herein for a human IgG1 Fc region or domain, as can be determined by one of skill in the art. In one embodiment, the Fc region or domain comprising such sequences exhibits one or more Fc activities, e.g., binding affinity for Fc receptors or effector functions, such as ADCC or CDC. In particular embodiments, the Fc region or domain comprising such a sequence exhibits reduced Fc activity, e.g., reduced binding affinity to Fc receptors or reduced effector functions, such as ADCC or CDC. In particular embodiments, the Fc region or domain comprising such a sequence exhibits enhanced FcRn activity, e.g., enhanced half-life.
Other non-limiting examples of Fc region or domain modifications are provided in Ghetie et al, 1997,Nat Biotech.15:637-40; duncan et al, 1988,Nature 332:563-564; lund et al, 1991,J.Immunol 147:2657-2662; lund et al, 1992,Mol Immunol 29:53-59; alegre et al, 1994,Transplantation 57:1537-1543; hutchins et al, 1995,Proc Natl.Acad Sci U S A92:11980-11984; jefferis et al, 1995,Immunol Lett.44:111-117; lund et al, 1995,Faseb J9:115-119; jefferis et al, 1996,Immunol Lett 54:101-104; lund et al, 1996,J Immunol157:4963-4969; armour et al 1999,Eur J Immunol 29:2613-2624; idusogie et al, 2000,JImmunol 164:4178-4184; reddy et al, 2000,J Immunol 164:1925-1933; xu et al, 2000,Cell Immunol 200:16-26; idusogie et al, 2001,J Immunol 166:2571-2575; shields et al 2001,J Biol Chem 276:6591-6604; jefferis et al, 2002,Immunol Lett 82:57-65; presta et al, 2002,Biochem Soc Trans 30:487-490); U.S. Pat. nos. 5,624,821;5,885,573;5,677,425;6,165,745;6,277,375;5,869,046;6,121,022;5,624,821;5,648,260;6,528,624;6,194,551;6,737,056;6,821,505;6,277,375;8,163,882;7,355,008;7,960,512;8,039,592;8,039,359;8,101,720;7,214,775;7,682,610;7,741,442; U.S. patent publication No.2004/0002587 and PCT patent publication WO 94/29351; WO 99/58372; WO 00/42072; WO 04/029207; WO 04/099249; WO 04/063151.
In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG1Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat. In particular embodiments, the modified (e.g., mutated) human IgG1Fc region or domain further comprises the non-naturally occurring amino acids 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
In certain embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG2 Fc region or domain comprising non-naturally occurring amino acids equivalent to 234A, 235Q, and 322Q numbered for the human IgG1Fc region or domain by the EU index as set forth in Kabat, as can be determined by one of skill in the art. In certain embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG2 Fc region or domain comprising non-naturally occurring amino acids equivalent to 234A, 235Q, 322Q, 252Y, 254T, and 256E numbered for the human IgG1Fc region or domain by the EU index as set forth in Kabat, as can be determined by one of skill in the art.
In certain embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG3 Fc region or domain comprising non-naturally occurring amino acids equivalent to 234A, 235Q, and 322Q numbered for the human IgG1 Fc region or domain by the EU index as set forth in Kabat, as can be determined by one of skill in the art. In certain embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG3 Fc region or domain comprising non-naturally occurring amino acids equivalent to 234A, 235Q, 322Q, 252Y, 254T, and 256E numbered for the human IgG1 Fc region or domain by the EU index as set forth in Kabat, as can be determined by one of skill in the art.
In certain embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG4 Fc region or domain comprising non-naturally occurring amino acids equivalent to 234A, 235Q, and 322Q numbered for the human IgG1 Fc region or domain by the EU index as set forth in Kabat, as can be determined by one of skill in the art. In certain embodiments, the antibodies described herein comprise a modified (e.g., mutated) human IgG4 Fc region or domain comprising non-naturally occurring amino acids equivalent to 234A, 235Q, 322Q, 252Y, 254T, and 256E numbered for the human IgG1 Fc region or domain by the EU index as set forth in Kabat, as can be determined by one of skill in the art.
In particular embodiments, the antibodies described herein comprise VL and VH CDR sequences shown in table 1 and a modified (e.g., mutated) human IgG1 Fc region or domain, wherein the modified (e.g., mutated) human IgG1 Fc region or domain comprises non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat.
In preferred embodiments, the antibodies described herein comprise VL and VH CDR sequences shown in table 1 and a modified (e.g., mutated) human IgG1 Fc region or domain, wherein the modified (e.g., mutated) human IgG1 Fc region or domain comprises non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat.
Thus, in one aspect, provided herein is an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT comprising:
(i) VL comprising a sequence having SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; (ii) VH comprising amino acid sequences having SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, and 322Q, as numbered by the EU index as set forth in Kabat. In a specific embodiment, the antibodies or antigen binding fragments thereof provided herein are antibodies.
Thus, in another aspect, provided herein is an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT comprising:
(i) VL comprising a sequence having SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; (ii) VH comprising amino acid sequences having SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat. In a specific embodiment, the antibodies or antigen binding fragments thereof provided herein are antibodies.
In another aspect, provided herein is an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT comprising: (i) VL comprising the amino acid sequence: DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSASYR YSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVEIK (SEQ ID NO: 17), wherein X K1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, X K2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, X K3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Amino acids having charged or acidic side chains and X K6 Is an amino acid having an aromatic side chain; and (ii) VH comprising the amino acid sequence: QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIARIY PGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGVYY FDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, 322Q, as numbered by the EU index as set forth in Kabat, and further preferably comprising 252Y, 254T, and 256E. In a specific embodiment, the antibodies or antigen binding fragments thereof provided herein are antibodies.
In another aspect, provided herein is an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT comprising: i) VL comprising SEQ ID NO: 13. 14, 15 or 16 and ii) VH comprising the amino acid sequence set forth in SEQ ID NO: 8. 9, 10, 11 or 12; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, 322Q, and preferably further comprising 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat. In a specific embodiment, the antibodies or antigen binding fragments thereof provided herein are antibodies.
In another aspect, provided herein is an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT comprising: i) VL comprising SEQ ID NO:14 and ii) a VH comprising the amino acid sequence set forth in SEQ ID NO:10, and a polypeptide comprising the amino acid sequence shown in seq id no; and (iii) a modified (e.g., mutated) human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, 322Q, and preferably further comprising 252Y, 254T, and 256E, as numbered by the EU index as set forth in Kabat. In a specific embodiment, the antibodies or antigen binding fragments thereof provided herein are antibodies.
In particular embodiments, the antibodies provided herein comprise a heavy chain comprising the following amino acid sequences
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSG
NTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTT
VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO：21)。
In particular embodiments, the antibodies provided herein comprise a light chain comprising the amino acid sequence: DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 22).
In particular embodiments, the antibodies provided herein comprise a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSG
NTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTT
VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP
CPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21); and a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSG
VPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVF
IFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
in particular embodiments, provided herein are antibodies comprising: (i) a heavy chain comprising the amino acid sequence:
(SEQ ID NO: 19) wherein the leader sequence is shown in bold italics, the variable region (VH) is shown in italics and the constant region is shown underlined. In addition, mutations in the constant region (compared to wild-type human IgG 1) are shown in double underlined; and
(ii) A light chain comprising the amino acid sequence:
wherein the leader sequence is shown in bold italics, the variable region (VL) is shown in italics and the constant region is shown underlined.
In particular embodiments, an anti-KIT antibody described herein does not bind to (e.g., has undetectable binding to) any human Fc-gamma receptor (fcγr receptor). In particular embodiments, an anti-KIT antibody described herein does not bind to (e.g., has undetectable binding to) human fcyri. In particular embodiments, an anti-KIT antibody described herein does not bind to (e.g., has undetectable binding to) human fcyriia. In particular embodiments, an anti-KIT antibody described herein does not bind to human fcyriib (e.g., has undetectable binding to fcyriib). In particular embodiments, the anti-KIT antibodies described herein do not bind to (e.g., have undetectable binding to) human fcyriiia. In particular embodiments, an anti-KIT antibody described herein does not bind to (e.g., has undetectable binding to) human fcyriiib.
In particular embodiments, the anti-KIT antibodies described herein comprise a modified (e.g., mutated) human IgG constant region (e.g., a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 constant region) and have enhanced binding (e.g., at least 2-fold, 5-fold, 10-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5000-fold, or 10000-fold higher binding affinity) to human neonatal Fc receptor (FcRn) relative to a corresponding antibody having the same variable region sequence, but having an unmodified (wild-type) human IgG constant region. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 20nM at pH 6.0. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 2nM at pH 6.0. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 1nM at pH 6.0. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 500nM at pH 6.0. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 400pM at pH 6.0. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 200nM at pH 7.2. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 150nM at pH 7.2. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 100nM at pH 7.2. In specific embodiments, an anti-KIT antibody described herein binds to FcRn with a KD of less than 80nM at pH 7.2.
In particular embodiments, the anti-KIT antibodies described herein comprise a modified (e.g., mutated) human IgG constant region (e.g., a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 constant region) and exhibit antibody-independent cell-mediated cytotoxicity (ADCC). In particular embodiments, the anti-KIT antibodies described herein comprise a modified (e.g., mutated) human IgG constant region (e.g., a modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 constant region) and exhibit reduced (e.g., at least 10% lower, at least 20% lower, at least 30% lower, at least 40% lower, at least 50% lower, at least 60% lower, at least 70% lower, at least 80% lower, at least 90% lower, at least 95% lower, or at least 99% lower) ADCC relative to a corresponding antibody having the same variable region sequence but with an unmodified (wild-type) human IgG constant region.
In particular embodiments, the anti-KIT antibodies described herein comprise modified (e.g., mutated) human IgG constant regions (e.g., modified (e.g., mutated) human IgG1, igG2, igG3, or IgG4 constant regions) and exhibit reduced (e.g., at least 10% lower, at least 20% lower, at least 30% lower, at least 40% lower, at least 50% lower, at least 60% lower, at least 70% lower, at least 80% lower, at least 90% lower, at least 95% lower, or at least 99% lower) cytokine (e.g., IFN- γ, IL-1β, IL-2, IL-6, IL-8, IL-10, and/or TNF- α) production relative to a corresponding antibody having the same variable region sequence but having an unmodified (wild-type) human IgG constant region.
In certain aspects, anti-KIT antibodies or antigen-binding fragments thereof have been described for use in methods of preventing, treating, or controlling chronic prurigo, or they may be readily obtained using methods known in the art, e.g., see section 5.2 below.
In a particular aspect, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to the D4 domain of human KIT and KIT, e.g., the D5 region of human KIT. In another specific embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to KIT, e.g., the D5 domain of human KIT, with less affinity than to KIT, e.g., the D4 domain of human KIT. In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to KIT with a higher affinity than to KIT, e.g., the D5 domain of human KIT, e.g., the D4 domain of human KIT, e.g., a higher affinity of at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, 500-fold, or 1000-fold, as determined by methods known in the art, e.g., ELISA or biacore assays.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to KIT, e.g., the D4 or D4/D5 region of human KIT, and has at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold higher affinity for KIT antigen consisting essentially of only the D4 domain than KIT antigen consisting essentially of only the D5 domain.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to a KIT polypeptide (e.g., the D4 region of human KIT), the EC thereof 50 The (half maximal effective concentration) value is about 50nM, 10nM, 500pM, 300pM, 200pM, 100pM, or 50pM or less, as determined by assays described in the art, as determined by ELISA.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to a KIT polypeptide (e.g., the D4 region of human KIT), the EC thereof 50 Values of about 200pM or 150pM or less, as determined by assays described in the art, such as ELISA or FACs using CHO-WT-KIT cells (CHO cells engineered to recombinantly express wild-type human KIT).
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein can be capable of an IC of about 600pM or less 50 (50% inhibition concentration) values block KIT phosphorylation.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of internalizing or enhancing KIT receptor by, e.g., at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% relative to internalization in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art. In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of internalizing a KIT receptor by, e.g., at least about 25% or 35%, optionally to about 75%, relative to internalization in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art. In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of internalizing or enhancing KIT receptor by at least about 1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold relative to internalization in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art. Techniques for quantification or visualization of cell surface receptors are well known in the art and include a variety of fluorescent and radioactive techniques. For example, one method involves incubating the cells with a radiolabeled anti-receptor antibody. Alternatively, the natural ligand of the receptor may be conjugated to a fluorescent molecule or radiolabel and incubated with the cell. Other receptor internalization assays are well known in the art and are described, for example, in Jisenez et al, biochemical Pharmacology,1999,57:1125-1131; bernhagen et al, nature Medicine,2007,13:587-596; and Conway et al, J.cell Physiol.,2001,189:341-55.
In particular embodiments, a renewal is used in the present context, relative to a renewal in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT)The anti-KIT antibodies, or antigen-binding fragments thereof, used in the methods provided herein are capable of eliciting or enhancing KIT receptor turnover, e.g., by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%, as assessed by methods described herein or known to those of skill in the art (e.g., pulse-chase assays). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of eliciting or enhancing KIT receptor turnover by at least about 25% or 35%, optionally to about 75%, relative to turnover in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art (e.g., pulse-chase assays). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of eliciting or enhancing KIT receptor turnover by at least about 1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold relative to turnover in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art (e.g., pulse-chase assays). Methods for determining receptor turnover are well known in the art. For example, one can use 35 S-EXPRESS protein labeling mixtures (NEG 772, NEN Life Science Products) pulse labeled KIT-expressing cells, wash and track with unlabeled vehicle for a period of time, then immunoprecipitate protein lysates from the labeled cells using anti-KIT antibodies, and separate and visualize by SDS-PAGE (e.g., exposure to PhosphoImager screen (Molecular Dynamics), scan using Typhoon8600 scanner (Amersham) and analyze using ImageQuant software (Molecular Dynamics) (see, e.g., chan et al, development,2004, 131:5551-5560).
In a specific embodimentIn embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of causing or enhancing KIT receptor degradation by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% relative to degradation in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art (e.g., pulse-chase assays). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of causing or enhancing KIT receptor degradation by at least about 25% or 35%, optionally to about 75%, relative to degradation in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art (e.g., pulse-chase assays). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is capable of causing or enhancing KIT receptor degradation by at least about 1-fold, 1.2-fold, 1.3-fold, 1.4-fold, 1.5-fold, 2-fold, 2.5-fold, 3-fold, 3.5-fold, 4-fold, 4.5-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold relative to degradation in the presence of an unrelated antibody (e.g., an antibody that does not immunospecifically bind to KIT), as assessed by methods described herein or known to those of skill in the art (e.g., pulse-chase assays). Techniques for determining or monitoring ubiquitination and/or degradation (e.g., kinetics or degradation rate) of cell surface receptors are well known in the art and include a variety of fluorescence and radioactivity techniques (see, e.g., international patent application publication No. wo 2008/153926 A2). For example, the use of radiolabeled ligands, such as 125 One or more pulse-chase experiments of I-SCF to quantitatively measure the degradation of KIT.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein does not bind to an extracellular ligand binding site of KIT, e.g., an SCF binding site of KIT. In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein does not inhibit ligand binding to KIT, e.g., does not inhibit KIT ligand (e.g., SCF) binding to KIT, as determined by methods described in the art, e.g., ELISA. In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein does not completely inhibit or partially inhibit binding to a ligand of KIT, e.g., does not completely inhibit or partially inhibit binding to a KIT ligand of KIT (e.g., SCF), as determined by methods described in the art, e.g., ELISA or FACS (fluorescence activated cell sorting).
In particular aspects, an anti-KIT antibody (e.g., a human or humanized antibody) for use in the methods provided herein is an inhibitory antibody, i.e., an antibody that inhibits (e.g., partially inhibits) KIT activity, i.e., one or more KIT activities. In specific embodiments, partial inhibition of KIT activity results in, for example, about 25% to about 65% or 75% inhibition. In particular embodiments, partial inhibition of KIT activity results in, for example, about 35% to about 85% or 95% inhibition. Non-limiting examples of KIT activity include KIT dimerization, KIT phosphorylation (e.g., tyrosine phosphorylation), KIT downstream signaling (e.g., stat, AKT, MAPK or Ras signaling), induction or enhancement of gene transcription (e.g., c-Myc), induction or enhancement of cell proliferation or cell survival. In particular embodiments, an antibody described herein inhibits KIT phosphorylation (e.g., ligand-induced phosphorylation).
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits KIT tyrosine phosphorylation in the cytoplasmic domain of KIT.
In another specific embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits cell proliferation, e.g., mast cell proliferation or eosinophil proliferation. In another embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits cell survival, e.g., mast cell survival or eosinophil survival. In certain aspects, the inhibition of cell proliferation, e.g., mast cell proliferation or eosinophil proliferation, is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.
In another specific embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits mast cell activation or eosinophil activation. In certain aspects, mast cell activation or activity or inhibition of eosinophil activation or activity is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits eosinophil or mast cell degranulation (see, e.g., staats et al 2012, med. Chem. Commun.,2013,4:88-94; and Ochkur et al 2012, j. Immunol. Methods, 384:10-20). In certain aspects, eosinophil or mast cell degranulation is inhibited by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.
In another specific embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits mast cell vehicle release. In certain aspects, the mast cell vehicle release is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. Assays have been described that measure mast cell activity, such as release of a vehicle from mast cell cultures, such as rodents and human mast cell cultures (see, e.g., kuehn et al, "Measuring Mast Cell Mediator Release," in Current Protocols in Immunology, unite 7.38.1-7.38.9,November 2010 (John Wiley & Sons, inc.). In certain aspects, CD34 peripheral blood progenitor cells or mast cell lines, such as HMC-1 or human LAD2 mast cell lines, may be used in these assays to determine the effect of anti-KIT antibodies on mast cells.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein induces apoptosis, e.g., mast cell apoptosis or eosinophil apoptosis. In another specific embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein induces cell differentiation, e.g., mast cell differentiation.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein may achieve any one of the following: a decrease in eosinophil number and/or activity, a decrease in mast cell proliferation, a decrease in mast cell number or amount, an inhibition or decrease in mast cell activity, a decrease in mast cell-induced production or release of inflammatory factors, a decrease in inflammatory factor release, a restoration of mast cell homeostasis, a decrease in mast cell migration, a decrease in mast cell adhesion, an inhibition or decrease in mast cell recruitment by eosinophils, and an inhibition or decrease in antigen-mediated degranulation of mast cells.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits KIT activity but does not inhibit KIT dimerization. In another specific embodiment, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits KIT activity and does not inhibit ligand binding to KIT, e.g., does not inhibit KIT ligand (e.g., SCF) binding to KIT, but does inhibit KIT dimerization.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits KIT activity, such as ligand-induced tyrosine phosphorylation of KIT cytoplasmic domains, by about 25% to about 65% or 75%, as determined by cell-based phosphorylation assays well known in the art, e.g., cell-based phosphorylation assays described herein. In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein inhibits KIT activity, such as ligand-induced tyrosine phosphorylation of KIT cytoplasmic domains, by about 35% to about 85% or 95%, as determined by cell-based phosphorylation assays well known in the art, e.g., cell-based phosphorylation assays described herein.
In specific embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is administered at a 50% Inhibitory Concentration (IC) of less than about 600pM, or less than about 500pM, or less than about 250pM 50 ) Inhibition of KIT activity, such as ligand-induced tyrosine phosphorylation of KIT cytoplasmic domains, as determined by cell-based phosphorylation assays well known in the art, e.g., cell-based phosphorylation assays described herein. In a specific embodiment, the IC 50 Less than about 550pM or 200pM. In a specific embodiment, the IC 50 In the range of about 50pM to about 225pM, or in the range of 100pM to about 600 pM. In a specific embodiment, the IC 50 In the range of about 50pM to about 550pM, or about 50pM to about 600pM, or about 150pM to about 550 pM.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein (i) immunospecifically binds to a KIT polypeptide comprising the D4 and/or D5 region of human KIT, (ii) inhibits KIT phosphorylation (e.g., tyrosine phosphorylation) and (iii) does not completely inhibit or partially inhibit KIT ligand (e.g., SCF) binding to KIT. In another embodiment, such an antibody does not inhibit KIT dimerization. In another embodiment, such antibodies can be recombinantly expressed by CHO cells at an average titer of at least 0.5 μg/mL, e.g., at least 1.0 μg/mL. In other embodiments, such antibodies comprise non-immunogenic VH and VL domains, e.g., VH and VL domains that do not contain T cell epitopes.
In other specific embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein immunospecifically binds to a monomeric form of KIT (e.g., human KIT). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to a monomeric form of KIT (e.g., human KIT). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to a dimeric form of KIT (e.g., human KIT).
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein does not bind to a monomeric form of KIT and specifically binds to a dimeric form of KIT or a multimeric form of KIT. In certain embodiments, the antibody has a higher affinity for KIT monomers than for KIT dimers. In certain embodiments, the antibody has a higher affinity for KIT monomers than for KIT multimers.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to a naturally occurring isoform or variant of KIT in an animal, preferably a human, that may be isolated from an animal (e.g., monkey, mouse, goat, donkey, dog, cat, rabbit, pig, rat, human, frog, or bird). In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof. In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof and does not specifically bind to non-human KIT (e.g., monkey, mouse, goat, donkey, dog, cat, rabbit, pig, rat, or bird) or fragment thereof. In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof and does not specifically bind to murine KIT. In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof (e.g., the D4 region of human KIT) and specifically binds to canine (dog) and non-human primate (e.g., monkey) KIT. In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof (e.g., the D4 region of human KIT) and specifically binds to canine (dog) and non-human primate (e.g., monkey) KIT, but does not specifically bind to murine or rat KIT or fragment thereof (e.g., the D4 region of murine KIT).
In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof (e.g., the D4 region of human KIT) and specifically binds to canine (dog), feline (cat) and cynomolgus KIT, but does not specifically bind to murine or rat KIT or fragment thereof (e.g., the D4 region of murine KIT).
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to human KIT or fragment thereof (e.g., the D4 region of human KIT) and specifically to canine (dog), feline (cat) and cynomolgus KIT with higher avidity (e.g., at least 0.5-fold, 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, or 10-fold) than murine or rat KIT or fragment thereof (e.g., the D4 region of murine KIT).
In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to the extracellular domain of human KIT comprising a mutation, e.g., a somatic mutation, such as a mutation in exon 9 of human KIT in which the Ala and Tyr residues at positions 502 and 503 are repeated (see, e.g., marcia et al, (2000) am.j. Pathol.156 (3): 791-795; and Debiec-Rychter et al, (2004) European Journal of cancer.40:689-695, all of which are incorporated herein by reference in their entirety, and describe KIT mutations).
In certain embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein specifically binds to the extracellular domain of glycosylated human KIT. In certain embodiments, the antibodies described herein, or antigen-binding fragments thereof, bind to 2 different glycosylated forms of the extracellular domain of human KIT. For example, 2 human KIT forms with different molecular weights have been observed by immunoblotting, indicating different glycosylation patterns.
In certain embodiments, the antibodies described herein can specifically bind to both forms of human KIT, which have different glycosylation patterns, e.g., one pattern is more glycosylated than the other. In some embodiments, the antibodies described herein, or antigen-binding fragments thereof, bind to the extracellular domain of human KIT.
In particular embodiments, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is a bivalent, monospecific antibody, wherein it has 2 antigen-binding regions (e.g., 2 identical antigen-binding regions) and both antigen-binding regions specifically bind to the same antigen KIT (e.g., human KIT). In certain embodiments, the antigen binding region comprises VH and VL CDRs as shown in table 1. In a specific embodiment, the antigen binding region comprises a VH comprising the amino acid sequence of SEQ ID NO:8-12, comprising the amino acid sequence set forth in any one of SEQ ID NOs: 13-16. In certain aspects, an anti-KIT antibody or antigen-binding fragment thereof for use in the methods provided herein is not a bispecific antibody.
In particular embodiments, the anti-KIT antibodies for use in the methods provided herein are Fab fragments that immunospecifically bind to a KIT polypeptide, such as the D4 region of KIT. In specific embodiments, the antibodies for use in the methods described herein are monoclonal antibodies or isolated monoclonal antibodies. In another specific embodiment, the antibody for use in the methods described herein is a humanized monoclonal antibody. In particular embodiments, the antibodies for use in the methods described herein are recombinant antibodies, e.g., recombinant human antibodies, recombinant humanized antibodies, or recombinant monoclonal antibodies. In certain embodiments, antibodies for use in the methods described herein contain non-human amino acid sequences, e.g., non-human CDRs or non-human (e.g., non-human primate) framework residues.
In particular embodiments provided herein, recombinant antibodies, such as antibodies expressed using recombinant expression vectors transfected into host cells, antibodies isolated from recombinant, combinatorial antibody libraries, or antibodies prepared, expressed, produced, or isolated by any other means, including, for example, via synthesis, genetic engineering of DNA sequences encoding human immunoglobulin sequences, or splicing of human immunoglobulin gene sequences with other such sequences, can be isolated, produced, expressed, or produced by recombinant means. In certain embodiments, the amino acid sequences of these recombinant antibodies have been modified such that the amino acid sequences of these antibodies, e.g., VH and/or VL regions, are non-naturally occurring sequences within a germline repertoire of biological antibodies, e.g., within a murine or human germline repertoire. In particular embodiments, the recombinant antibody can be obtained by assembling some sequence fragments naturally occurring in an organism (e.g., primate, such as human) into a composite sequence of the recombinant antibody, wherein the composite sequence does not naturally occur within the organism (e.g., primate, such as human).
Antibodies for use in the methods provided herein include immunoglobulin molecules of any type (e.g., igG, igE, igM, igD, igA and IgY), class (e.g., igG1, igG2, igG3, igG4, igA1, and IgA 2), or subclass. In particular embodiments, the antibodies for use in the methods provided herein are IgG antibodies (e.g., human IgG antibodies) or classes (e.g., human IgG1 or IgG 4) or subclasses thereof. In another specific embodiment, the antibody for use in the methods described herein is an IgG1 (e.g., human IgG1 (isotype a, z or f)) or IgG4 antibody. In certain embodiments, the antibody for use in the methods described herein is a whole or whole antibody, e.g., a whole or whole humanized, human, or complex human antibody.
In particular aspects, antibodies provided herein comprise antibody light and heavy chains, e.g., isolated light and heavy chains. In particular embodiments, the light chain of the antibodies described herein is a kappa light chain, relative to a light chain. In another specific embodiment, the light chain of an antibody described herein is a lambda light chain. In another embodiment, the light chain of an antibody described herein is a human kappa light chain or a human lambda light chain. In specific embodiments, the antibodies described herein comprise a human light chain constant region. Non-limiting examples of human light chain constant region sequences have been described in the art, see, for example, U.S. Pat. No.5,693,780 and Kabat et al (1991) Sequences of Proteins of Immunological Interest, 5 th edition, U.S. Pat. No. of Health and Human Services, NIH Publication No.91-3242.
In particular embodiments, the heavy chain of an antibody described herein can be an alpha (α), delta (δ), epsilon (epsilon), gamma (gamma), or mu (mu) heavy chain, relative to the heavy chain. In another specific embodiment, the heavy chain of the antibody may comprise a human α (α), δ (δ), ε (ε), γ (γ), or μ (μ) heavy chain. In particular embodiments, the antibodies described herein comprise a human heavy chain constant region (e.g., a human IgG constant region, e.g., a human IgG1, igG2, igG3, or IgG4 constant region). Non-limiting examples of human heavy chain constant region sequences have been described in the art, see, for example, U.S. Pat. No.5,693,780 and Kabat et al (1991) Sequences of Proteins of Immunological Interest, 5 th edition, U.S. Pat. No. of Health and Human Services, NIH Publication No.91-3242. In particular embodiments, the antibodies described herein comprise a modified (e.g., mutated) human Fc region or domain (e.g., a modified (e.g., mutated) human IgG1 Fc region or domain, a modified (e.g., mutated) human IgG2 Fc region or domain, a modified (e.g., mutated) human IgG3 Fc region or domain, or a modified (e.g., mutated) human IgG4 Fc region or domain.
Antibodies provided herein can include antibody fragments that retain the ability to specifically bind to an antigen, e.g., a KIT epitope (e.g., a KIT epitope within a KIT polypeptide comprising the D4 region of human KIT). In specific embodiments, fragments include Fab fragments (antibody fragments containing an antigen-binding domain and comprising light and heavy chain portions bridged by disulfide bonds (i.e., the CHI domains of VH and heavy chains); fab' (antibody fragment containing a single antigen-binding domain comprising Fab and the other part of the heavy chain to the hinge region); f (ab') 2 (2 Fab 'molecules linked by interchain disulfide bonds in the heavy chain hinge region; the Fab' molecules may be directed against the same or different epitopes); dual specificity Fab (Fab molecules with 2 antigen binding domains, each of which may be directed against a different epitope); single chain Fab chain comprising variable region, also called sfv (by 10-25 amino acidsVariable, antigen-binding determining regions of single light and heavy chains of an antibody linked together; disulfide-linked Fv or dsFv (variable, antigen-binding determining regions of a single light and heavy chain of an antibody linked together by disulfide bonds); caramelised VH (variable, antigen-binding determining region of a single heavy chain of an antibody, wherein some amino acids at the VH interface are those found in naturally occurring camelid antibody heavy chains); dual specificity sfvs (sFv or dsFv molecules with 2 antigen-binding domains, each of which may be directed against a different epitope); diabodies (dimeric sFvs are formed when the VH domain of a first sFv is assembled with the VL domain of a second sFv and the VL domain of the first sFv is assembled with the VH domain of the second sFv; the 2 antigen-binding regions of the diabodies may be directed against the same or different epitopes); and a three-chain antibody (a trimeric sFv formed in a manner similar to a diabody, but wherein three antigen-binding domains are produced in a single complex; the three antigen-binding domains may be directed against the same or different epitopes). Antibodies provided herein may also include one or more CDR sequences of an antibody. When two or more CDR sequences are present, the CDR sequences may be linked together in the framework. In certain embodiments, the antibody comprises a single-chain Fv ("scFv"). scFv are antibody fragments comprising VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Typically, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the scFv to form the desired structure for antigen binding. For a review of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, volume 113, rosenburg and Moore eds. Springer-Verlag, new York, pages 269-315 (1994). Without being bound by any particular theory, fv molecules may be capable of penetrating tissue due to their smaller size. Complete antibodies can be cleaved enzymatically by pepsin to produce F (ab') 2 Fragments, or may be enzymatically cleaved by papain to produce 2 Fab fragments. The antibody or antigen binding fragment thereof may also be linked to other binding entities, such as SCF binding sequences or "trap".
In certain embodiments, the anti-KIT antibody for use in the methods described herein is a human, composite human, or humanized monoclonal antibody. In particular embodiments, the antibodies for use in the methods described herein are engineered antibodies, e.g., antibodies produced by recombinant methods. In particular embodiments, the antibodies described herein are humanized antibodies comprising one or more non-human (e.g., rodent or murine) CDRs and one or more human Framework Regions (FR), and optionally human heavy chain constant regions and/or light chain constant regions. In particular embodiments, the antibodies described herein comprise one or more primate (or non-human primate) framework regions. In specific embodiments, the antibodies described herein do not include a non-human primate framework region.
Antibodies for use in the methods provided herein can include antibodies comprising chemical modifications, e.g., antibodies that have been chemically modified, e.g., by covalently linking any type of molecule to the antibody. For example, but not by way of limitation, anti-KIT antibodies may be glycosylated, acetylated, pegylated, phosphorylated, or amidated, may be derivatized via a protecting/blocking group, or may further comprise a cellular ligand and/or other proteins or peptides (e.g., heterologous proteins or peptides), and the like. For example, the antibodies provided herein can be chemically modified, e.g., by glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, attachment to a cellular ligand or other protein, and the like. In addition, the anti-KIT antibodies described herein may contain one or more atypical amino acids. In some embodiments, provided herein are conjugates comprising an anti-KIT antibody or antigen-binding fragment thereof linked to another agent (e.g., a therapeutic agent). These antibody conjugates can also be used in methods or uses described in the present disclosure.
In one embodiment, an anti-KIT antibody for use in the methods provided herein is a naked antibody that is not linked, fused or conjugated (e.g., artificially linked, fused or conjugated) to another molecule, peptide or polypeptide (e.g., a heterologous polypeptide). In particular embodiments, the anti-KIT antibody for use in the methods provided herein is not an antibody-drug conjugate. In particular embodiments, the anti-KIT antibody for use in the methods provided herein is not a fusion protein. In particular embodiments, the anti-KIT antibodies described herein do not comprise any atypical amino acids.
5.2 antibody production
Antibodies (e.g., human or humanized antibodies) (or antigen-binding fragments thereof) that immunospecifically bind to a KIT antigen described herein can be produced by any method known in the art for antibody synthesis, e.g., by chemical synthesis or by recombinant expression techniques. Unless otherwise indicated, the methods described herein employ techniques conventional in the relevant arts of molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and within the skill of the art. These techniques are described in the references cited herein and are fully explained in the literature. See, e.g., maniatis et al (1982) Molecular Cloning: A Laboratory Manual, cold Spring Harbor Laboratory Press; sambrook et al (1989), molecular Cloning: A Laboratory Manual, 2 nd edition, cold Spring Harbor Laboratory Press; sambrook et al (2001) Molecular Cloning: A Laboratory Manual, cold Spring Harbor Laboratory Press, cold Spring Harbor, NY; ausubel et al Current Protocols in Molecular Biology, john Wiley & Sons (1987 and annual revisions); current Protocols in Immunology, john Wiley & Sons (1987 and annual revisions); gait (Main plaited) (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press; eckstein (Main plaited) (1991) Oligonucleotides and Analogues: A Practical Approach, IRL Press; birren et al (Main plaited) (1999) Genome Analysis: A Laboratory Manual, cold Spring Harbor Laboratory Press.
For example, humanized antibodies can be produced using a variety of techniques known in the art, including (but not limited to) CDR-grafting (european patent No. EP 239,400; international patent publication No. WO 91/09967; and U.S. Pat. Nos. 5,225,539, 5,530,101 and 5,585,089), veneering (veneering) or surface remodeling (resurfacing) (European patent Nos. EP 592,106 and EP 519,596;Padlan,1991,Molecular Immunology28 (4/5): 489-498; studnica et al, 1994,Protein Engineering 7 (6): 805-814); and Roguska et al, 1994, PNAS 91:969-973), chain shuffling (U.S. Pat. No.5,565,332) and, for example, U.S. Pat. No.6,407,213, U.S. Pat. No.5,766,886, WO 9317105, tan et al, J.Immunol.169:1119 25 (2002), caldas et al, protein Eng.13 (5): 353-60 (2000), morea et al, methods 20 (3): 267 79 (2000), baca et al, J.biol. Chem.272 (16): 10678-84 (1997), roguska et al, protein Eng.9 (10): 895 904 (1996), couto et al, cancer Res.55 (23 support): 5973s-5977s (1995), couto et al, cancer Res.55 (8): 1717-22 (1995), pedfu JS 150 (409-10): 35 (1994), and Biol et al, 1993). See also U.S. patent publication No. US 2005/0042664Al (24.2.2005), incorporated herein by reference.
Monoclonal antibodies can be prepared using a variety of techniques known in the art, including the use of hybridoma, recombinant, and phage display techniques, or combinations thereof. For example, hybridoma techniques can be used, including those known in the art and described, for example, in Harlow et al, antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2 nd edition 1988); monoclonal antibodies are generated by those taught in Hammerling et al, in Monoclonal Antibodies and T-Cell Hybridomas563 681 (Elsevier, N.Y., 1981). The term "monoclonal antibody" as used herein is not limited to antibodies produced by hybridoma technology. For example, monoclonal antibodies can be produced by recombinant techniques, e.g., by expression of the recombinant monoclonal antibodies by a host cell, such as a mammalian host cell.
Methods for generating and screening specific antibodies using hybridoma technology are conventional and well known in the art. For example, in a hybridoma method, a mouse or other suitable host animal, such as sheep, goat, rabbit, rat, hamster, or cynomolgus monkey, is immunized to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization (e.g., the extracellular domain of human KIT). Alternatively, lymphocytes can be immunized in vitro. Lymphocytes are then fused with myeloma cells using a suitable fusion agent, such as polyethylene glycol, to form hybridoma cells (Goding, monoclonal Antibodies: principles and Practice, pages 59-103 (Academic Press, 1986)). In addition, RIMMS (multiple site repeat immunization) techniques may be used to immunize animals (Kilptrack et al, 1997Hybridoma 16:381-9, incorporated herein by reference).
Non-limiting examples of myeloma cell lines include murine myeloma lines such as those derived from MOPC-21 and MPC-11 mouse tumors from Salk Institute Cell Distribution Center, san Diego, calif., USA, and SP-2 or X63-Ag8.653 cells from American type culture Collection (American Type Culture Collection), rockville, MD, USA. Human myeloma and mouse-human hybrid myeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor, J.Immunol.,133:3001 (1984); brodeur et al, monoclonal Antibody Production Techniques and Applications, pages 51-63 (Marcel Dekker, inc., new York, 1987)). [00157]Antibodies described herein include antibody fragments that recognize a specific KIT antigen, and the antibodies described herein can be produced by any technique known to those of skill in the art. For example, it is possible to use enzymes such as papain (to produce Fab fragments) or pepsin (to produce F (ab') 2 Fragments) of immunoglobulin molecules to produce Fab and F (ab') 2 Fragments. The Fab fragment corresponds to one of the two identical arms of the antibody molecule and contains the complete light chain paired with the VH and CHI domains of the heavy chain. F (ab') 2 The fragment contains two antigen-binding arms of the antibody molecule linked by disulfide bonds in the hinge region.
In one aspect, to generate an intact antibody, PCR primers comprising VH or VL nucleotide sequences, restriction sites, and flanking sequences protecting the restriction sites can be used to clonally amplify VH or VL sequences from a template, e.g., scFv. The PCR amplified VH domain may be cloned into a vector expressing a VH constant region, and the PCR amplified VL domain may be cloned into a vector expressing a VL constant region, e.g., a human kappa or lambda constant region, using cloning techniques known to those skilled in the art. The VH and VL domains may also be cloned into a vector expressing the necessary constant regions. The heavy chain and light chain transfer vectors are then co-transfected into a cell line using techniques known to those skilled in the art to produce a stable or transient cell line expressing full length antibodies, e.g., igG.
Single domain antibodies, e.g., antibodies lacking a light chain, may be produced by methods well known in the art. See Riechmann et al 1999, J.Immunol.231:25-38; nuttall et al, 2000, curr.Pharm.Biotechnol.l (3): 253-263; muylederman, 2001, J. Iotechnol.74 (4): 277302; U.S. Pat. No.6,005,079; and International patent publication Nos. WO 94/04678, WO 94/25591 and WO 01/44301.
5.3 compositions
Provided herein are compositions, such as pharmaceutical compositions, comprising one or more anti-KIT antibodies (e.g., humanized antibodies) or antigen-binding fragments thereof, for use in the methods described herein. In particular aspects, the compositions described herein may be used in vitro, in vivo, or ex vivo. In particular embodiments, provided herein are pharmaceutical compositions comprising an anti-KIT antibody (e.g., humanized antibody) or antigen-binding fragment thereof for use in the methods described herein and a pharmaceutically acceptable carrier or excipient.
As used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the federal or a state government or listed in the U.S. pharmacopeia, european pharmacopeia, or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
Antibodies of the desired purity may be prepared by combining the antibodies with an optional physiologically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences (1990) Mack Publishing Co., easton, pa., remington: the Science and Practice of Pharmacy, 21 st edition (2006) Lippincott Williams)&Wilkins, baltimore, MD) are mixed to prepare therapeutic formulations provided herein containing one or more antibodies (e.g., humanized antibodies) in the form of a lyophilized formulation or an aqueous solution. At the dosages and concentrations employed, the acceptable carrier, excipient or stabilizer is non-toxic to the recipient and includes Buffers such as phosphates, citrates and other organic acids; and/or nonionic surfactants, e.g. TWEEN TM 、PLURONICS TM Or polyethylene glycol (PEG).
Formulations, such as those described herein, may also contain more than one active compound (e.g., a molecule, e.g., one or more antibodies described herein) as necessary for the particular indication to be treated. In certain embodiments, the formulation comprises an antibody provided herein and one or more active compounds having complementary activities that do not adversely affect each other. These molecules are suitably present in combination in an amount effective for the intended purpose.
The formulation to be used for in vivo administration may be sterile. This is easily accomplished by filtration through, for example, a sterile filtration membrane.
In particular aspects, the pharmaceutical compositions provided herein contain a therapeutically effective amount of one or more anti-KIT antibodies (e.g., humanized antibodies) provided herein, and optionally one or more other prophylactic or therapeutic agents, in a pharmaceutically acceptable carrier. These pharmaceutical compositions are useful in preventing, treating, controlling or ameliorating chronic prurigo or one or more symptoms thereof.
Pharmaceutical carriers suitable for administration of the antibodies provided herein include any of these carriers known to those skilled in the art as suitable for the particular form of administration.
In addition, the antibodies described herein may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients (e.g., one or more other prophylactic or therapeutic agents).
The composition may contain one or more anti-KIT antibodies provided herein. In one embodiment, the antibodies are formulated in sterile solutions or suspensions for parenteral administration as suitable pharmaceutical preparations, such as solutions, suspensions, powders or elixirs. In one embodiment, the antibodies are formulated into pharmaceutical formulations suitable for oral administration, such as solutions, suspensions, tablets, dispersible tablets, pills, capsules, powders, sustained release formulations or elixirs, as well as transdermal patch formulations and dry powder inhalers.
In these compositions, one or more of the antibodies (or conjugates thereof) provided herein are admixed with a suitable pharmaceutical carrier. The concentration of the one or more antibodies in the composition may be effective, for example, to deliver an amount that treats, prevents, protects, or controls chronic prurigo or one or more symptoms thereof once administered.
In one embodiment, the composition is formulated for single dose administration. To formulate the composition, the weight fraction of the compound is dissolved, suspended, dispersed or otherwise admixed in a carrier of choice at an effective concentration such that the condition being treated is alleviated, prevented, or one or more symptoms are ameliorated.
In certain aspects, the antibodies (e.g., humanized antibodies) (or antibody-drug conjugates thereof) provided herein are included in a pharmaceutically acceptable carrier in an amount effective to exert a therapeutically useful effect without undesired side effects or with minimal or negligible undesired side effects on the patient being treated. The therapeutically effective concentration can be determined empirically by testing the compound in vitro and in vivo systems using conventional methods, and then extrapolated therefrom for the dosage of humans.
The concentration of antibody in the pharmaceutical composition will depend, for example, on the physicochemical characteristics of the antibody, the dosage schedule and the amount to be administered, as well as other factors known to those skilled in the art. In certain aspects, the concentration of antibody-drug conjugate in the pharmaceutical composition will depend, for example, on the physicochemical characteristics of the antibody and/or drug, the dosage schedule, and the amount to be administered, among other factors known to those of skill in the art.
In one embodiment, a therapeutically effective dose produces a serum concentration of antibody of about 0.1ng/ml to about 50-100 μg/ml. In another embodiment, the pharmaceutical composition provides a dose of about 0.001mg to about 2000mg of antibody per kilogram of body weight for administration over a period of time, e.g., daily, weekly, every 2 weeks, or every 3, 4, or 8 weeks. Pharmaceutical dosage unit forms can be prepared to provide from about 0.01mg to about 2000mg, and in one embodiment, from about 10mg to about 500mg of the combination of antibodies and/or other optional essential ingredients per dosage unit form.
In specific embodiments, the antibody-drug conjugates described herein are administered at an effective dose of about 1 to 100mg of antibody-drug conjugate per kilogram of body weight for a period of time, e.g., daily, weekly, every 2 weeks, or every 3 weeks.
The anti-KIT antibodies described herein may be administered at once, or may be divided into multiple smaller doses for administration at intervals. It will be appreciated that the exact dosage and duration of treatment will be related to the disease to be treated and may be determined empirically using known testing protocols or by extrapolation of in vivo or in vitro test data. It should be noted that the concentration and dosage values may also vary with the severity of the condition to be alleviated. It will also be appreciated that the particular dosing regimen may be adjusted over time for any particular subject according to individual needs and the professional judgment of the person administering or supervising the administration of the compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions.
Once the antibodies are mixed or added, the resulting mixture may be a solution, suspension, emulsion, or the like. The form of the resulting mixture depends on several factors, including the intended form of administration and the solubility of the compound in the chosen carrier or vehicle. The effective concentration is sufficient to ameliorate the symptoms of the disease, disorder or condition to be treated and can be determined empirically.
The pharmaceutical compositions described herein are provided in unit dosage forms, such as sterile parenteral (e.g., intravenous) solutions or suspensions, containing a suitable amount of a compound or a pharmaceutically acceptable derivative thereof for administration to humans and animals, such as mammals (e.g., cats or dogs). Pharmaceutical compositions are also provided in unit dosage forms, such as tablets, capsules, pills, powders, granules and oral solutions or suspensions, as well as oil-in-water emulsions, containing suitable amounts of the compound or a pharmaceutically acceptable derivative thereof for administration to humans and animals, such as mammals (e.g., cats or dogs). In one embodiment, the antibody is formulated and administered in unit dosage form or in multiple dosage form. A unit dosage form as used herein refers to physically separate units suitable as are known in the art for human and animal subjects and packaged separately. Each unit dose in combination with the desired pharmaceutical carrier, vehicle or diluent contains a predetermined amount of antibody sufficient to produce the desired therapeutic effect. Examples of unit dosage forms include ampoules and syringes and individually packaged tablets or capsules. The unit dosage form may be administered in portions or multiple times. Multiple dosage forms are multiple identical unit dosage forms packaged in a single container to be administered in separate unit dosage forms. Examples of multi-dose forms include vials, tablet or capsule bottles or pints or gallon bottles. Thus, a multi-dose form is a plurality of unit doses that are not separated in the package.
In certain embodiments, one or more anti-KIT antibodies described herein are in a liquid pharmaceutical formulation. Liquid pharmaceutically administrable compositions may be prepared, for example, by dissolving, dispersing or otherwise mixing an active compound as defined above and optionally a pharmaceutical adjuvant in a carrier such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, or the like, thereby forming a solution or suspension. The pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting agents, emulsifying agents, solubilizing agents, and pH buffering agents, among others, if desired.
The actual methods of preparing these dosage forms are known or will be apparent to those skilled in the art; see, e.g., remington's Pharmaceutical Sciences (1990) Mack publishing co., easton, PA; remington, the Science and Practice of Pharmacy, 21 st edition (2006) Lippincott Williams & Wilkins, baltimore, md.
Dosage forms or compositions containing antibodies in the range of 0.005% to 100% can be prepared with the remainder consisting of a nontoxic carrier. Methods for preparing these compositions are known to those skilled in the art.
In one embodiment, parenteral administration is also contemplated herein, characterized by subcutaneous, intramuscular, or intravenous injection. The injection may be prepared in conventional form as a liquid solution or suspension, as a solid form suitable for solution or suspension in a liquid prior to injection, or as an emulsion. The injectables, solutions and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol or ethanol. In addition, the pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubilizing agents, and other such agents, if desired. Other routes of administration may include epidural, enteral, intracerebral, nasal, intraarterial, intracardiac, intraosseous infusion, intrathecal, and intraperitoneal administration. In specific embodiments, an anti-KIT antibody, antigen-binding fragment, or pharmaceutical composition described herein is administered intravenously. In specific embodiments, an anti-KIT antibody, antigen-binding fragment, or pharmaceutical composition described herein is administered subcutaneously.
Formulations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products ready for incorporation with solvents just prior to use, such as lyophilized powders, including subcutaneous tablets, sterile suspensions ready for injection, sterile dry insoluble products ready for incorporation with vehicles just prior to use, and sterile emulsions. The solution may be aqueous or non-aqueous.
If administered intravenously, suitable carriers include physiological saline or Phosphate Buffered Saline (PBS), and solutions containing thickening and solubilizing agents, such as dextrose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.
Pharmaceutically acceptable carriers for use in parenteral formulations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents and other pharmaceutically acceptable substances.
The pharmaceutical carrier further comprises ethanol, polyethylene glycol, and propylene glycol for a water miscible vehicle; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.
Illustratively, intravenous or intra-arterial infusion of sterile aqueous solutions containing the active compound is an effective administration form. Another embodiment is a sterile aqueous or oily solution or suspension containing the active material for injection as needed to produce the desired pharmacological effect.
The anti-KIT antibodies described herein may be suspended in micronized or other suitable form. The form of the resulting mixture depends on several factors, including the intended form of administration and the solubility of the compound in the chosen carrier or vehicle. The effective concentration is sufficient to ameliorate the symptoms of the condition and can be determined empirically.
In other embodiments, the pharmaceutical formulation is a freeze-dried powder that can be reconstituted for administration as a solution, emulsion, and other mixture. They may also be reconstituted and formulated as a solid or gel.
Lyophilized powders are prepared by dissolving the antibodies provided herein in a suitable solvent. In some embodiments, the lyophilized powder is sterile. The solvent may contain excipients that improve the stability or other pharmacological components of the powder or reconstituted solution prepared from the powder. Excipients that may be used include, but are not limited to, glucose, sorbitol, fructose, corn syrup, xylitol, glycerol, glucose, sucrose, or other suitable agents. The solvent may also contain a buffer, such as citrate, sodium or potassium phosphate, or other such buffers known to those skilled in the art to be at about neutral pH in one embodiment. Subsequent sterile filtration of the solution and subsequent freeze-drying under standard conditions, known to those skilled in the art, provides the desired formulation. In one embodiment, the resulting solution is dispensed into vials for lyophilization. Each vial will contain a single dose or multiple doses of the compound. The lyophilized powder may be stored under suitable conditions, such as at about 4 ℃ to room temperature.
Reconstitution of such lyophilized powders with water for injection provides a formulation for use in parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable carrier. The exact amount depends on the compound selected. These amounts may be determined empirically.
The antibodies described herein can be formulated for topical or surface application, such as topical application to skin and mucous membranes, such as the eye, in the form of gels, creams and lotions, and for application to the eye or for intracisternal or intraspinal application. Topical administration for transdermal delivery, for ocular or mucosal administration, or for inhalation therapy is contemplated. Nasal solutions of the active compounds alone or in combination with other pharmaceutically acceptable excipients may also be administered.
Antibodies and other compositions provided herein can also be formulated to target specific tissues, receptors, or other areas of the subject's body to be treated. A variety of these targeting methods are well known to those skilled in the art. All such targeting methods for use in the compositions of the invention are contemplated herein. For non-limiting examples of targeting methods, see, e.g., U.S. Pat. nos. 6,316,652, 6,274,552, 6,271,359, 6,253,872, 6,139,865, 6,131,570, 6,120,751, 6,071,495, 6,060,082, 6,048,736, 6,039,975, 6,004,534, 5,985,307, 5,972,366, 5,900,252, 5,840,674, 5,759,542, and 5,709,874. In some embodiments, an anti-KIT antibody described herein is targeted (or otherwise administered) to bone marrow. In some embodiments, an anti-KIT antibody described herein is targeted (or otherwise administered) to the gastrointestinal tract. In some embodiments, an anti-KIT antibody described herein is targeted (or otherwise administered) to the brain. In particular embodiments, the anti-KIT antibodies described herein are capable of crossing the blood brain barrier.
In specific embodiments, an anti-KIT antibody described herein is targeted (or otherwise administered) to an ocular tissue or organ. In particular aspects, compositions comprising an anti-KIT antibody described herein may be targeted to ocular tissues or organs as eye drops or gels. In particular aspects, compositions comprising an anti-KIT antibody described herein may be targeted to the ear.
Provided herein are pharmaceutical packages or kits comprising one or more containers filled with one or more components of the pharmaceutical compositions described herein, such as one or more antibodies provided herein. Optionally, associated with such containers may be a notice in the form prescribed by a government agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency for human administration.
For KIT inhibitors other than antibodies, it is contemplated that they may also be formulated in pharmaceutical formulations along with pharmaceutically acceptable carriers suitable for storage, handling, and/or administration.
5.4 doses and administration
According to the methods provided herein for treating chronic prurigo, the dosage and frequency of administration of an anti-KIT antibody described herein, or a pharmaceutical composition thereof, to a subject (e.g., mammal, such as a human, dog, or cat) in need thereof will be effective while minimizing side effects. The exact dosage of an anti-KIT antibody or pharmaceutical composition thereof described herein to be administered to a particular subject may be determined based on factors related to the subject in need of treatment. Factors that may be considered include the severity of the disease state, the general health of the subject, the age and weight of the subject, diet, time and frequency of administration, combination with other therapeutic agents or drugs, response sensitivity, and tolerance/response to therapy. The dosage and frequency of administration of the anti-KIT antibodies or pharmaceutical compositions thereof described herein may be adjusted over time to provide adequate levels of anti-KIT antibodies or to maintain a desired effect.
The exact dosage to be used in the formulation will also depend on the route of administration and the severity of the condition or disease, and should be determined according to the judgment of the practitioner and each patient's circumstances.
In certain aspects, for an anti-KIT antibody described herein, the dose administered to a patient to prevent, protect, control, or treat chronic prurigo is typically 0.1mg/kg to 100mg/kg of patient body weight. In a specific embodiment, the dose administered to a patient to prevent, protect, control or treat chronic prurigo is about 3mg/kg patient body weight.
Generally, human antibodies have a longer half-life in humans than antibodies derived from other species due to an immune response to foreign polypeptides. Thus, lower doses of human antibodies and less frequent administration are generally possible. Furthermore, the dosage and frequency of administration of the antibodies described herein can be reduced by modification, such as, for example, lipidation, to increase absorption and tissue penetration of the antibodies.
In one embodiment, about 0.001mg/kg (mg antibody per kg subject body weight) to about 500mg/kg of an anti-KIT antibody described herein is administered to prevent, protect, control, or treat chronic prurigo. In another embodiment, about 0.3mg/kg (mg antibody per kg subject body weight) of an anti-KIT antibody described herein is administered to prevent, protect, control, or treat chronic prurigo. In another embodiment, about 1mg/kg (mg antibody per kg subject body weight) of an anti-KIT antibody described herein is administered to prevent, protect, control, or treat chronic prurigo. In another embodiment, about 3mg/kg (mg antibody per kg subject body weight) of an anti-KIT antibody described herein is administered to prevent, protect, control, or treat chronic prurigo. In another embodiment, about 9mg/kg (mg antibody per kg subject body weight) of an anti-KIT antibody described herein is administered to prevent, protect, control, or treat chronic prurigo.
In some embodiments, an effective amount of an antibody provided herein is about 0.01mg to about 1,000mg. In particular embodiments, an "effective amount" or "therapeutically effective amount" of an anti-KIT antibody described herein refers to an amount of an anti-KIT antibody described herein sufficient to effect at least one, two, three, four, or more of the following effects: a reduction or improvement in the severity of chronic prurigo and/or one or more symptoms associated therewith; a reduction in the duration of one or more symptoms associated with the chronic prurigo; prevention of recurrence of one or more symptoms of chronic prurigo and/or one or more symptoms associated therewith; reduction in hospitalization of the subject; reduction in hospitalization length; inhibition (e.g., partial inhibition) of the development of chronic prurigo and/or one or more symptoms associated therewith; prevention of the development or onset of one or more symptoms associated with chronic prurigo; a decrease in the concentration of one or more inflammatory mediators (e.g., cytokines or interleukins) in a biological sample (e.g., plasma, serum, cerebrospinal fluid, urine, or any other biological fluid) of a subject suffering from chronic prurigo; and improvement in quality of life as assessed by methods well known in the art, e.g., questionnaires. In some embodiments, an "effective amount" as used herein also refers to an amount of an antibody described herein that achieves the indicated result (e.g., inhibition of one or more KIT biological activities of a cell, such as inhibition of cell proliferation).
In some embodiments, an anti-KIT antibody described herein is administered as needed, e.g., weekly, biweekly (i.e., once every two weeks), monthly, bi-monthly, tri-monthly, etc.
In some embodiments, a single dose of one or more anti-KIT antibodies described herein is administered to a patient to hinder, prevent, control, treat, and/or ameliorate chronic prurigo.
In particular embodiments, an anti-KIT antibody or pharmaceutical composition thereof is administered to a subject in accordance with the methods provided herein for treating chronic prurigo, wherein the anti-KIT antibody or pharmaceutical composition is administered for a period of time followed by a discontinuation of the administration of the anti-KIT antibody or pharmaceutical composition for a period of time (i.e., not administered for a period of time).
The methods provided herein comprise administering an anti-KIT antibody by any suitable route. Non-limiting examples of routes of administration include parenteral administration, e.g., subcutaneous, intramuscular, or intravenous administration, epidural administration, enteral administration, intracerebral administration, nasal administration, intraarterial administration, intracardiac administration, intraosseous infusion, intrathecal administration, and intraperitoneal administration. The methods provided herein include an administration route that targets brain, ocular tissue or organ, spinal cord, or ear or auricular tissue. In particular aspects, the methods provided herein include an administration route that targets the nervous system, e.g., the central nervous system.
In particular embodiments, the methods provided herein comprise administering an anti-KIT antibody via a route suitable for crossing the blood brain barrier.
For KIT inhibitors other than antibodies, it is contemplated that the above description of dosages and administration also applies to the extent deemed appropriate by the treating physician.
6. Examples
The examples are provided herein by way of illustration and not by way of limitation.
6.1 example 1
Nucleic acid molecules encoding the Fc mutated H (heavy chain) and L (light chain) amino acid sequences of the anti-KIT antibodies (SEQ ID NO:23 and SEQ ID NO:24, respectively, see Table 6 below) were cloned directly into expression vectors. The construct was confirmed by sequencing and the vector was transfected into CHO cells. Stable transfection to establish cell lines expressing the antibodies is performed and subsequent drug selection is performed. The best expression line was selected based on IgG titers in the supernatant above background and amplified in the presence of drug selection and used at each stageThe QKe system screens IgG expression.
Table 6: DNA sequences encoding the heavy and light chain sequences of an anti-KIT antibody.
After removal of the leader sequence, the resulting antibody was denoted antibody mAb1. The heavy and light chain amino acid sequences (after removal of the leader sequence) of the antibodies are shown in table 7 below.
Table 7: full length heavy and light chain amino acid sequences of mAb 1.
The Fc domain of the heavy chain of the antibody comprises non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T and 256E as numbered by the EU index as set forth in Kabat. The 6 non-naturally occurring amino acids are shown in bold and underlined text in the above sequence.
It was determined that mAb1 did not cause significant degranulation of FcgRI-expressing human mast cells compared to the corresponding antibody ("mAbc") with a wild-type (non-mutated) IgG1 Fc domain (as shown by the% release of β -hexosaminidase from human mast cells in culture). The release of β -hexosaminidase from human mast cells in culture (in the presence of ifnγ) was reduced by more than 50% by mAb1 compared to mAbc. In addition, mAb1 did not show significant Fc receptor-dependent KIT agonist activity (as determined by KIT phosphorylation) compared to mAbc, even though crosslinked on THP-1 cells. Fc receptor-dependent KIT agonist activity (as determined by KIT phosphorylation using cross-linked Fc receptors) was reduced by more than 50% with mAb1 compared to mAbc.
6.2 example 2
Healthy volunteers were provided with a single infusion of either 0.3, 1, 3 or 9mg/kg mAb1 or placebo. Using detection of both alpha and beta forms of tryptase Total plasma tryptase levels were measured by assay. For each group, the tryptase values were normalized to 100% for the pretreatment values and to 0% for the lower assay limit (1 ng/mL). The mean and mean standard error are plotted against dose.
Figures 2A-2E show results demonstrating that mAb1 inhibits plasma tryptase in a dose-dependent manner.
6.3 example 3
A single dose of mAb1 resulted in an extended decrease in tryptase values below the quantitative level determined (1 ng/mL). Optionally, values below the detection level are plotted as 0.5 ng/mL. Absolute plasma tryptase values are shown.
Figures 3A and 3B show results demonstrating that a single dose of mAb1 provides durable tryptase inhibition at both 3mg/kg and 9 mg/kg.
6.4 example 4
Healthy volunteers were provided with a single infusion of either 0.3, 1, 3 or 9mg/kg mAb1 or placebo. The assay developed internally measures plasma levels of Stem Cell Factor (SCF), the unique ligand of the c-KIT/CD117 receptor, by using the Meso Scale Diagnostics (MSD) platform. SCF plasma levels rise in a dose-dependent manner, consistent with the allosteric blockade of KIT receptors by SCF. The mean and mean standard error are plotted against dose.
Figure 4 shows the results, which demonstrate that mAb1 induced a dose-dependent increase in plasma SCF levels.
6.5 example 5
M-07e cells were serum starved, then pretreated with i) mAb1, ii) a corresponding antibody with the same variable region sequence, but with unmutated (wild-type) human IgG1 sequence ("mAbc"), iii) isotype control antibody, or iv) a KIT-targeted small molecule kinase inhibitor (imatinib), and then stimulated with Stem Cell Factor (SCF). Phosphorylation was assessed by western blotting.
Fig. 5 shows the results from one experiment. The data shown represents 3 independent experiments. IgG = human IgG1 isotype control antibody; p-KIT = tyrosine phosphorylated KIT; p-AKT = phosphorylated AKT; p-ERK = phosphorylated ERK1/2. Overall, these results indicate that mAb1 is a more potent inhibitor of SCF-induced activation of wild-type KIT and downstream intracellular signaling pathways than the small molecule KIT inhibitors tested.
6.6 example 6
The effect of mAb1 and imatinib on SCF-dependent M-07e cell proliferation was also identified.
M-07e cells were serum starved, then pretreated with i) mAb1, ii) a corresponding antibody with the same variable region sequence, but with unmutated (wild-type) human IgG1 sequence ("mAbc") or iii) imatinib, and then stimulated with SCF. Cells were incubated at 37℃for 6 days.
The results are shown in fig. 6. The data represent 3 independent experiments and are expressed as the mean and mean standard error of three technical replicates.
Both mAb1 and mAbc showed similar dose-dependent inhibition of M-07e cell proliferation with average IC50 values (±sem; n=5; fig. 6) of 1.11±0.15nM and 1.12±0.22nM, respectively. By comparison, imatinib was less effective in inhibiting SCF-dependent M-07e cell proliferation with an average IC50 of 228±39nM (±sem) from 3 independent experiments. The antagonist activity of mAb1 was comparable to mAbc, indicating that introducing mutations into the Fc region of mAb1 did not affect its ability to inhibit KIT.
6.7 example 7
The binding affinity of mAb1 to recombinant human Fc-gamma receptor (fcγr) and human neonatal Fc receptor (FcRn) was identified and a balanced KD value was generated.
UsingQKe instrument binding to recombinant human Fc receptor was measured by biofilm interferometry. Histidine-tagged Fc receptors were captured on anti-pentahis biosensors and then exposed to serial dilutions of mAbc or mAb1 for 2-3 minutes followed by a dissociation step in the range of 5-10 minutes. Curve fitting was performed using instrument analysis software. Binding to FcRn was performed by a binding and dissociation step at pH 6.0 and pH 7.2.
The results are shown in fig. 7 and 8A-8N, which demonstrate that mAbc binds to fcγri (kd=3.39 nM) with high affinity, has medium affinity for fcγriiia (kd=127 nM) and weak affinity for fcγriia (kd=391 nM) and fcγriiib (kd=816 nM). No binding to fcyriib was observed. In contrast, mAb1 (concentration=500 nM) was detected to have no binding to any recombinant human fcγr.
Binding to human neonatal Fc receptor (FcRn) was tested at physiological pH 7.2 and at pH 6.0 mimicking endocytic conditions. At pH 7.2, no binding of mAbc to FcRn was observed, whereas mAb1 bound with moderate affinity (kd=77.1 nM). Mabc bound to FcRn with moderate affinity (kd=211 nM) at pH 6.0. However, mab1 bound to FcRn with significantly higher affinity and showed very slow dissociation (kd=0.38 nM) at pH 6.0.
These data indicate that Fc mutations in mAb1 eliminate fcγr interactions while enhancing in vitro interactions with FcRn.
6.8 example 8
Antibody-dependent cell-mediated cytotoxicity (ADCC) activity of mAb1 was assessed using a commercially available ADCC Reporter Bioassay kit (Promega Corporation, madison, WI). ADCC Reporter Bioassay engineered Jurkat cells stably expressing the FcgammaRIIIa receptor V158 (high affinity) variant and nuclear factors driving the activated T cell (NFAT) response element of firefly luciferase expression were used as effector cells. Binding of fcyriiia receptor on the surface of effector cells to the Fc effector portion of antibodies that bind to antigens on target cells will result in cross-linking and activation of fcyriiia signaling, which will cause expression of the NFAT reporter. To evaluate the ability of mAb1 and mAbc to induce ADCC, M-07e cells expressing KIT, transfected CHO cells (CHO-WT KIT) and human small cell lung cancer H526 cells were used as target cells. In addition, untransfected CHO cells that do not express KIT were used as control target cells. ADCC is considered to have been activated if a dose-dependent increase in the resulting reporter signal is observed.
As shown in fig. 9, mAbc elicited ADCC response to KIT-expressing M-07e target cells, whereas mAb1 or isotype control did not. Similar results were observed with CHO-WT KIT and H526 cells. Overall, these data demonstrate that mAb1, unlike mAbc, does not elicit ADCC response against the examined KIT expressing target cells.
6.9 example 9
Fresh whole blood was incubated overnight with 40nM huIgG1 isotype control or 0.02, 0.2 or 40nM mAb1 in solution or in dry coating as indicated. Phytohemagglutinin (PHA) (10. Mu.g/mL) and Lipopolysaccharide (LPS) (10. Mu.g/mL) were used as positive controls. Plasma samples were harvested and stored frozen at-80 ℃. Cytokine levels were determined by using a multiplexed laser bead technique (Multiplexing Laser Bead Technology) implemented by Eve Technologies (Calgary, alberta, canada).
Fig. 10 shows the results, wherein the cytokine concentration of each donor is expressed as the average of two replicates. The mean cytokine concentration of all donors (n=6) was plotted using an error bar representing SEM. The results show that very little specific cytokine induction as a whole was observed by mAb1 relative to the human IgG1 isotype control.
6.10 example 10
The human patient develops chronic itching and multiple prurigo skin lesions. Patients were diagnosed with chronic prurigo and mAb1 was administered intravenously. Patients were monitored by clinical evaluation before, during and after the response to treatment.
6.11 example 11
Described herein are open label phase 1 single dose study protocols to evaluate the safety, pharmacokinetics, and pharmacodynamics of mAb1 as an adjunct therapy in patients with cold-contact urticaria, symptomatic skin scars, or cholinergic urticaria. The study type was interventional (clinical trial). The intervention mode is assigned to a single group.
The present study is an open-label phase 1 study that evaluates the safety, pharmacokinetics and pharmacodynamics of a single dose of mAb1 in patients with symptomatic cold-contact urticaria, symptomatic skin scars or cholinergic urticaria despite treatment with antihistamines. It is estimated that 10 patients with cold contact urticaria, 10 patients with symptomatic skin scars and 10 patients with cholinergic urticaria will be enrolled in three separate groups, totaling 30 patients. Prospective patients were screened by pre-recruited clinic testing and daily home diaries for 2 weeks. On day 1, a single dose of mAb1 was administered intravenously. Following treatment, the patient was followed for 12 weeks. Adult patients from 18 years to 75 years of age for all sexes are suitable for this study. Healthy volunteers were not received.
Primary outcome measure:
1. safety was assessed by the incidence and severity of adverse events (time frame: from day 1 to week 12). The safety of a single dose of mAb1 was determined by adverse events.
Secondary outcome measure:
1. for patients with cold-contact urticaria, the threshold temperature threshold (CTT) varies (time range: from day 1 to day 85). Such as by usingAs determined by excitation testing of (c) a critical temperature threshold over time relative to the baseLine change.
2. For patients with symptomatic skin scarification, a threshold change (time range: from day 1 to day 85) was elicited. Such as by usingAs determined by the excitation test of (c), the excitation threshold varies over time from baseline.
3. For patients with cholinergic urticaria, the baseline urticaria activity score elicits a change (UASprovo) (time frame: from day 1 to day 85). Changes and percentages of responders from baseline as measured by uasprova.
4. Changes from baseline (time frame: from day 1 to day 85) in Urticaria Control Test (UCT). For UCT and modified UCT, the change and percentage of responders from baseline.
5. Blood biomarkers (time range: from day 1 to day 85). Blood samples before and after treatment were collected and analyzed for stem cell factor changes.
6. Blood biomarkers (time range: from day 1 to day 85). Blood samples were collected before and after treatment and analyzed for tryptase changes.
7. Pharmacokinetic assessment (time frame: from day 1 to day 85). mAb1 concentration was measured.
8. Immunogenicity evaluation (time frame: from day 1 to day 85). The patient is monitored for the presence of anti-drug antibodies.
Key inclusion criteria:
1. diagnosis of cold contact urticaria, symptomatic skin scars or cholinergic urticaria that do not respond to antihistamines. Diagnosing for more than or equal to 3 months; although antihistamines are used simultaneously, symptoms of both urticaria (rubella) and itching/burning/pain sensations. During screening, in the clinic, patients must have a positive cold stimulus test for cold contact urticaria, positive for symptomatic skin scarificationAnd for cholinergic urticaria, the patient must have a positive pulse controlled muscle strength test (PCE) challenge test. The subject is at a stable antihistamine dose.
2. In addition to the diagnosis of cold contact urticaria, symptomatic skin scars, or cholinergic urticaria, no other risk factors will be introduced or other conditions will interfere with the study procedure as determined by the investigator based on the medical evaluation.
3. Male and female patients must use very effective contraception for at least 150 days from screening visit to receiving study treatment.
4. Willing and able to follow all study requirements and procedures, including completion of daily medication diaries and questionnaires.
Key exclusion criteria:
1. in addition to chronic urticaria, it is clear to diagnose urticaria or angioedema.
2. Prior biologic therapies (e.g., omalizumab, domino Li Youshan antibody, li Geli bead mab (ligelizumab)) were received within the last 3 months.
3. Treatment with immunosuppressants (e.g., systemic corticosteroids, cyclosporine, methotrexate, dapsone, cyclophosphamide, tacrolimus and mycophenolate mofetil, hydroxychloroquine, etc.) occurs over a 4 week or 5 half-life period.
4. Active covd-19 infection.
Infection with hiv, hepatitis b or hepatitis c.
There are other criteria for the treating physician to examine the candidate to confirm that it is appropriate for the study.
6.12 example 12
6.12.1 study background and summary
Chronic induced urticaria (CIndU) is characterized by Mast Cell (MC) -driven rubella that responds to cold in an initiator, such as cold in cold urticaria (cold) or skin scratch in symptomatic skin Scarification (SD). These diseases are often severe and debilitating, which can significantly affect patient longevity. MC require activation of their KIT receptors by stem cytokines to survive, proliferate and differentiate. MC loading is related to circulating tryptase (a protease secreted specifically by MC). mAb1 is a monoclonal anti-KIT antibody engineered to selectively inhibit Stem Cell Factor (SCF) -dependent KIT activation (see fig. 11). mAb1 demonstrated a significant dose-related reduction in circulating tryptase and was well tolerated overall in healthy volunteers. In this study, CIndU patients benefited from mAb1 treatment. The 1 SD patient enrolled in this study was also diagnosed with prurigo nodularis, which improved after treatment with mAb 1.
6.12.2 study design and method
In this ongoing open label phase 1b trial, coldU and SD patients refractory to antihistamine treatment received a single IV infusion of 3mg/kg mAb1 (as an additional treatment for H1-antihistamine) and were followed for 12 weeks. The patient was induced for cold and SD symptoms via a challenge test similar to the actual challenge. The main objective was to evaluate the safety/tolerability of mAb1 (adverse events and clinical laboratory tests). Secondary and research objectives include pharmacokinetic and pharmacodynamic assessments including changes in the threshold of stimulation from baseline, measurement of tryptase and stem cytokine levels, clinical outcome of activity (impact on urticaria symptoms, disease control, clinical response), quality of life assessment, and measurement of tissue mast cells by skin biopsy. Secondary goals included evaluating the effect of mAb1 on clinical activity and serum tryptase. The activity end point comprises the excitation test of disease severity /ColdU；/>SD), physician global assessment (Phys-GA) and patient global assessment (Pat-GA). Mean ± Standard Error (SE) of the challenge test, biomarker, and hematology are shown in fig. 12C-12D, fig. 14A-14C, fig. 15A-15D, and fig. 16A-16D, respectively. The number of skin MC assessed using non-diseased skin biopsies was counted by tryptase staining.
1 SD patient enrolled in this study was also diagnosed with prurigo nodularis.
The study was modified to add patient groups with cholinergic urticaria.
The study was conducted substantially in accordance with the clinical trial protocol described in example 11.
6.12.3 study State
20 patients received study drug (i.e., mAb 1) at 3mg/kg by a single intravenous infusion and were included in the safety analysis. 11 had ColdU and 9 had SD. One of the 9 SD patients also had prurigo nodularis. As assessed by the challenge threshold test, patients had high disease activity of CIndU. In a ColdU patient, the baseline critical temperature threshold is 18.9deg.C/66F (range: 5-27deg.C/41-80.6F). In SD patients, baselineThe threshold was 3.8 (range: 3-4) for 4 needles.
The 19 patients received full dose and were included in the activity analysis, including patients with both SD and prurigo nodularis. 14 of the 19 patients completed a 12 week observation period, including patients with both SD and prurigo nodularis; the 5-bit is in progress.
6.12.4 demographics and baseline disease characteristics
For identification of 20 patients, see table 8 below. All patients had prior antihistamine treatment.
Table 8: demographic and baseline disease characteristics
6.12.5 results of the study
As shown in fig. 12A-12D, a single dose of mAb1 (3 mg/kg) resulted in a rapid, pronounced and sustained response in cisdu patients refractory to antihistamines. In 95% (18/19) of patients (Cold patients)100% (10/10) (FIG. 12A) and 89% (8/9) (FIG. 12B) of the SD patients achieved a Complete Response (CR). Prior to having a prior(omalizumab) all 3 patients (1 ColdU patient and 2 SD patients) experienced, including 2 +.>Complete response was observed in refractory patients. Rapid response onset and sustained persistence following dose administration were observed. By week 1 and week 4, respectively, most of the ColdU and SD patients experienced a complete response. In the cold u patients who completed the 12 week follow-up period (i.e., 8 cold u patients and 6 SD patients), CR was maintained for a median duration of 77 days, and in the SD patients for 57 days (fig. 12C and 12D). Of the 19 patients, 1 (SD patient) underwent Partial Response (PR). Cr=negative challenge test at ∈4 ℃ (for cold) or 0 needle (for SD). Pr=to 4 ℃ (for ColdU) or ≡2 (for SD).
The improved disease activity as assessed by Phys-GA and Pat-GA was consistent with the complete response as measured by the challenge test (FIGS. 13A and 13B, for ColdU and SD patients, respectively).
As shown in fig. 14A and 14B, a single 3mg/kg mAb1 dose resulted in rapid, significant and durable depletion of skin MC (87% depletion, see fig. 14A) and inhibition of serum tryptase (fig. 14B), as measured by biopsy. MC and tryptase kinetics showed similar trends over time (fig. 14C). In addition, skin MC number correlated positively with serum tryptase levels (fig. 14D).
The kinetics of skin MC and serum tryptase depletion reflect the clinical activity of CIndU. In particular, the kinetics of skin MC and serum tryptase depletion reflect a decrease in the challenge threshold (FIGS. 15A-15D). The data confirm that serum tryptase levels are robust pharmacodynamic biomarkers for assessing MC burden and clinical activity in CIndU patients and potentially other diseases with mast cell driven conditions.
After a single dose of mAb1, patients with both SD and prurigo nodularis experienced a complete response to SD and significant prurigo nodularis improvement.
In addition, mAb1 demonstrated good safety and tolerability. mAb1 is generally well tolerated in the treated patients. The most common adverse events were changes in hair color (14/20 (70%)), infusion reactions (9/20 (45%)) and dysgeusia (8/20 (40%)). Changes in hair color (usually a small area of hair color lighten) and dysgeusia (usually a partial change in the ability to discriminate salty taste) are consistent with the inhibition of KIT signaling in other cell types and are expected to be fully reversible. Most adverse events were lighter. With longer observation period, the change in hair color was improved. Infusion reactions (often becoming urticaria and itching sensations) resolve spontaneously. A single severe infusion response of transient loss of consciousness occurred in patients with a history of syncope and was not due to MC activation, as measured by serum tryptase monitoring. The patient quickly recovers. Taste impairment is selective and transient. The hematology parameters generally remain within normal ranges (see fig. 16A-16D). A slight, transient and asymptomatic decrease in hemoglobin and White Blood Cell (WBC) parameters was noted (see fig. 16A-16D). However, there was no sign of clinically significant decrease in hematological parameters. This is an important finding for KIT inhibitors.
Overall, the data show that mAb1 demonstrates unprecedented MC depletion and a good safety profile, which provides significant potential as a cendu therapy for rapid, long lasting and meaningful alleviation, suggesting its potential to affect other diseases with mast cell involvement and providing an opportunity to evaluate MC involvement in a variety of diseases.
The scope of the invention is not limited by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.
All references cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual patent publication or patent application was specifically and individually indicated to be incorporated by reference in its entirety for all purposes.
Sequence listing
<110> Seldex medical Co Ltd
<120> treatment of chronic prurigo
<130> 12638-171-228
<140> TBA
<141>
<150> US 63/140,621
<151> 2021-01-22
<150> US 63/238,688
<151> 2021-08-30
<160> 40
<170> PatentIn version 3.5
<210> 1
<211> 972
<212> PRT
<213> person
<220>
<223> human KIT receptor
<400> 1
Met Arg Gly Ala Arg Gly Ala Trp Asp Phe Leu Cys Val Leu Leu Leu
1 5 10 15
Leu Leu Arg Val Gln Thr Gly Ser Ser Gln Pro Ser Val Ser Pro Gly
20 25 30
Glu Pro Ser Pro Pro Ser Ile His Pro Gly Lys Ser Asp Leu Ile Val
35 40 45
Arg Val Gly Asp Glu Ile Arg Leu Leu Cys Thr Asp Pro Gly Phe Val
50 55 60
Lys Trp Thr Phe Glu Ile Leu Asp Glu Thr Asn Glu Asn Lys Gln Asn
65 70 75 80
Glu Trp Ile Thr Glu Lys Ala Glu Ala Thr Asn Thr Gly Lys Tyr Thr
85 90 95
Cys Thr Asn Lys His Gly Leu Ser Asn Ser Ile Tyr Val Phe Val Arg
100 105 110
Asp Pro Ala Lys Leu Phe Leu Val Asp Arg Ser Leu Tyr Gly Lys Glu
115 120 125
Asp Asn Asp Thr Leu Val Arg Cys Pro Leu Thr Asp Pro Glu Val Thr
130 135 140
Asn Tyr Ser Leu Lys Gly Cys Gln Gly Lys Pro Leu Pro Lys Asp Leu
145 150 155 160
Arg Phe Ile Pro Asp Pro Lys Ala Gly Ile Met Ile Lys Ser Val Lys
165 170 175
Arg Ala Tyr His Arg Leu Cys Leu His Cys Ser Val Asp Gln Glu Gly
180 185 190
Lys Ser Val Leu Ser Glu Lys Phe Ile Leu Lys Val Arg Pro Ala Phe
195 200 205
Lys Ala Val Pro Val Val Ser Val Ser Lys Ala Ser Tyr Leu Leu Arg
210 215 220
Glu Gly Glu Glu Phe Thr Val Thr Cys Thr Ile Lys Asp Val Ser Ser
225 230 235 240
Ser Val Tyr Ser Thr Trp Lys Arg Glu Asn Ser Gln Thr Lys Leu Gln
245 250 255
Glu Lys Tyr Asn Ser Trp His His Gly Asp Phe Asn Tyr Glu Arg Gln
260 265 270
Ala Thr Leu Thr Ile Ser Ser Ala Arg Val Asn Asp Ser Gly Val Phe
275 280 285
Met Cys Tyr Ala Asn Asn Thr Phe Gly Ser Ala Asn Val Thr Thr Thr
290 295 300
Leu Glu Val Val Asp Lys Gly Phe Ile Asn Ile Phe Pro Met Ile Asn
305 310 315 320
Thr Thr Val Phe Val Asn Asp Gly Glu Asn Val Asp Leu Ile Val Glu
325 330 335
Tyr Glu Ala Phe Pro Lys Pro Glu His Gln Gln Trp Ile Tyr Met Asn
340 345 350
Arg Thr Phe Thr Asp Lys Trp Glu Asp Tyr Pro Lys Ser Glu Asn Glu
355 360 365
Ser Asn Ile Arg Tyr Val Ser Glu Leu His Leu Thr Arg Leu Lys Gly
370 375 380
Thr Glu Gly Gly Thr Tyr Thr Phe Leu Val Ser Asn Ser Asp Val Asn
385 390 395 400
Ala Ala Ile Ala Phe Asn Val Tyr Val Asn Thr Lys Pro Glu Ile Leu
405 410 415
Thr Tyr Asp Arg Leu Val Asn Gly Met Leu Gln Cys Val Ala Ala Gly
420 425 430
Phe Pro Glu Pro Thr Ile Asp Trp Tyr Phe Cys Pro Gly Thr Glu Gln
435 440 445
Arg Cys Ser Ala Ser Val Leu Pro Val Asp Val Gln Thr Leu Asn Ser
450 455 460
Ser Gly Pro Pro Phe Gly Lys Leu Val Val Gln Ser Ser Ile Asp Ser
465 470 475 480
Ser Ala Phe Lys His Asn Gly Thr Val Glu Cys Lys Ala Tyr Asn Asp
485 490 495
Val Gly Lys Thr Ser Ala Tyr Phe Asn Phe Ala Phe Lys Glu Gln Ile
500 505 510
His Pro His Thr Leu Phe Thr Pro Leu Leu Ile Gly Phe Val Ile Val
515 520 525
Ala Gly Met Met Cys Ile Ile Val Met Ile Leu Thr Tyr Lys Tyr Leu
530 535 540
Gln Lys Pro Met Tyr Glu Val Gln Trp Lys Val Val Glu Glu Ile Asn
545 550 555 560
Gly Asn Asn Tyr Val Tyr Ile Asp Pro Thr Gln Leu Pro Tyr Asp His
565 570 575
Lys Trp Glu Phe Pro Arg Asn Arg Leu Ser Phe Gly Lys Thr Leu Gly
580 585 590
Ala Gly Ala Phe Gly Lys Val Val Glu Ala Thr Ala Tyr Gly Leu Ile
595 600 605
Lys Ser Asp Ala Ala Met Thr Val Ala Val Lys Met Leu Lys Pro Ser
610 615 620
Ala His Leu Thr Glu Arg Glu Ala Leu Met Ser Glu Leu Lys Val Leu
625 630 635 640
Ser Tyr Leu Gly Asn His Met Asn Ile Val Asn Leu Leu Gly Ala Cys
645 650 655
Thr Ile Gly Gly Pro Thr Leu Val Ile Thr Glu Tyr Cys Cys Tyr Gly
660 665 670
Asp Leu Leu Asn Phe Leu Arg Arg Lys Arg Asp Ser Phe Ile Cys Ser
675 680 685
Lys Gln Glu Asp His Ala Glu Ala Ala Leu Tyr Lys Asn Leu Leu His
690 695 700
Ser Lys Glu Ser Ser Cys Ser Asp Ser Thr Asn Glu Tyr Met Asp Met
705 710 715 720
Lys Pro Gly Val Ser Tyr Val Val Pro Thr Lys Ala Asp Lys Arg Arg
725 730 735
Ser Val Arg Ile Gly Ser Tyr Ile Glu Arg Asp Val Thr Pro Ala Ile
740 745 750
Met Glu Asp Asp Glu Leu Ala Leu Asp Leu Glu Asp Leu Leu Ser Phe
755 760 765
Ser Tyr Gln Val Ala Lys Gly Met Ala Phe Leu Ala Ser Lys Asn Cys
770 775 780
Ile His Arg Asp Leu Ala Ala Arg Asn Ile Leu Leu Thr His Gly Arg
785 790 795 800
Ile Thr Lys Ile Cys Asp Phe Gly Leu Ala Arg Asp Ile Lys Asn Asp
805 810 815
Ser Asn Tyr Val Val Lys Gly Asn Ala Arg Leu Pro Val Lys Trp Met
820 825 830
Ala Pro Glu Ser Ile Phe Asn Cys Val Tyr Thr Phe Glu Ser Asp Val
835 840 845
Trp Ser Tyr Gly Ile Phe Leu Trp Glu Leu Phe Ser Leu Gly Ser Ser
850 855 860
Pro Tyr Pro Gly Met Pro Val Asp Ser Lys Phe Tyr Lys Met Ile Lys
865 870 875 880
Glu Gly Phe Arg Met Leu Ser Pro Glu His Ala Pro Ala Glu Met Tyr
885 890 895
Asp Ile Met Lys Thr Cys Trp Asp Ala Asp Pro Leu Lys Arg Pro Thr
900 905 910
Phe Lys Gln Ile Val Gln Leu Ile Glu Lys Gln Ile Ser Glu Ser Thr
915 920 925
Asn His Ile Tyr Ser Asn Leu Ala Asn Cys Ser Pro Asn Arg Gln Lys
930 935 940
Pro Val Val Asp His Ser Val Arg Ile Asn Ser Val Gly Ser Thr Ala
945 950 955 960
Ser Ser Ser Gln Pro Leu Leu Val His Asp Asp Val
965 970
<210> 2
<211> 11
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR1 of anti-KIT antibody (VL-CDR 1-AbM)
<400> 2
Lys Ala Ser Gln Asn Val Arg Thr Asn Val Ala
1 5 10
<210> 3
<211> 7
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR2 of anti-KIT antibody (VL-CDR 2-AbM)
<400> 3
Ser Ala Ser Tyr Arg Tyr Ser
1 5
<210> 4
<211> 9
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR3 of anti-KIT antibody (VL-CDR 3-AbM)
<400> 4
Gln Gln Tyr Asn Ser Tyr Pro Arg Thr
1 5
<210> 5
<211> 5
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR1 of anti-KIT antibody (VH-CDR 1)
<400> 5
Asp Tyr Tyr Ile Asn
1 5
<210> 6
<211> 17
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR2 of anti-KIT antibody (VH-CDR 2)
<400> 6
Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe Lys
1 5 10 15
Gly
<210> 7
<211> 7
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR3 of anti-KIT antibody (VH-CDR 3)
<400> 7
Gly Val Tyr Tyr Phe Asp Tyr
1 5
<210> 8
<211> 116
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain variable region of anti-KIT antibody (VH 1)
<400> 8
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Leu Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Ala Thr Leu Thr Ala Glu Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Arg Ser Glu Asp Ser Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 9
<211> 116
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain variable region of anti-KIT antibody (VH 2)
<400> 9
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Ala Thr Leu Thr Ala Glu Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 10
<211> 116
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain variable region of anti-KIT antibody (VH 3)
<400> 10
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 11
<211> 116
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain variable region of anti-KIT antibody (VH 4)
<400> 11
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Ala Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 12
<211> 116
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain variable region of anti-KIT antibody (VH 5)
<400> 12
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 13
<211> 107
<212> PRT
<213> artificial sequence
<220>
<223> light chain variable region of anti-KIT antibody (VL 1)
<400> 13
Asp Ile Val Met Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 14
<211> 107
<212> PRT
<213> artificial sequence
<220>
<223> light chain variable region of anti-KIT antibody (VL 2)
<400> 14
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 15
<211> 107
<212> PRT
<213> artificial sequence
<220>
<223> light chain variable region of anti-KIT antibody (VL 3)
<400> 15
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 16
<211> 107
<212> PRT
<213> artificial sequence
<220>
<223> light chain variable region of anti-KIT antibody (VL 4)
<400> 16
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 17
<211> 107
<212> PRT
<213> artificial sequence
<220>
<223> light chain variable region consensus sequences
<220>
<221> misc_feature
<222> (10)..(10)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aromatic or aliphatic hydroxyl side chains; or in another embodiment Xaa is the amino acid Phe or Ser
<220>
<221> misc_feature
<222> (46)..(46)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic or aliphatic hydroxyl side chains; or in another embodiment Xaa is the amino acid Ala or Ser
<220>
<221> misc_feature
<222> (63)..(63)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic or aliphatic hydroxyl side chains; or in another embodiment Xaa is the amino acid Thr or Ser
<220>
<221> misc_feature
<222> (80)..(80)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic or aliphatic hydroxyl side chains; or in another embodiment Xaa is the amino acid Ser or Pro
<220>
<221> misc_feature
<222> (85)..(85)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having charged or acidic side chains; or in another embodiment Xaa is the amino acid Asp or Thr
<220>
<221> misc_feature
<222> (87)..(87)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aromatic side chains; or in another embodiment Xaa is the amino acid Phe or Tyr
<400> 17
Asp Ile Val Met Thr Gln Ser Pro Ser Xaa Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Xaa Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Xaa Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Xaa
65 70 75 80
Glu Asp Phe Ala Xaa Tyr Xaa Cys Gln Gln Tyr Asn Ser Tyr Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
100 105
<210> 18
<211> 116
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain variable region consensus sequences
<220>
<221> misc_feature
<222> (11)..(11)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic side chains; or in another embodiment Xaa is the amino acid Leu or Val
<220>
<221> misc_feature
<222> (20)..(20)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic side chains; or in another embodiment Xaa is the amino acid Leu or Val
<220>
<221> misc_feature
<222> (38)..(38)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having polar or basic side chains; or in another embodiment Xaa is the amino acid Lys or Arg
<220>
<221> misc_feature
<222> (68)..(68)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic side chains; or in another embodiment Xaa is the amino acid Val or Ala
<220>
<221> misc_feature
<222> (70)..(70)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic side chains; or in another embodiment Xaa is the amino acid Leu or Ile
<220>
<221> misc_feature
<222> (73)..(73)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having acidic side chains; or in another embodiment Xaa is the amino acid Glu or Asp
<220>
<221> misc_feature
<222> (82)..(82)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having acidic or amide derivative side chains; or in another embodiment Xaa is the amino acid Gln or Glu
<220>
<221> misc_feature
<222> (91)..(91)
<223> Xaa can be any amino acid; or in a specific embodiment, it may be
Amino acids having aliphatic hydroxyl side chains; or in another embodiment Xaa is the amino acid Ser or Thr
<400> 18
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Xaa Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Xaa Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Xaa Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Xaa Thr Xaa Thr Ala Xaa Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Xaa Leu Ser Ser Leu Arg Ser Glu Asp Xaa Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser
115
<210> 19
<211> 464
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain sequence having mutation
<400> 19
Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly
1 5 10 15
Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
20 25 30
Pro Gly Ala Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe
35 40 45
Thr Asp Tyr Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
50 55 60
Glu Trp Ile Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn
65 70 75 80
Glu Lys Phe Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser
85 90 95
Thr Ala Tyr Met Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
100 105 110
Tyr Phe Cys Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly
115 120 125
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
130 135 140
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
145 150 155 160
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
165 170 175
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu
180 185 190
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
195 200 205
Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
210 215 220
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys
225 230 235 240
Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Gln Gly Gly Pro
245 250 255
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr
260 265 270
Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
275 280 285
Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
290 295 300
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
305 310 315 320
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
325 330 335
Tyr Lys Cys Gln Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys
340 345 350
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
355 360 365
Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
370 375 380
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
385 390 395 400
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu
405 410 415
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
420 425 430
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
435 440 445
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
450 455 460
<210> 20
<211> 234
<212> PRT
<213> artificial sequence
<220>
<223> light chain sequence
<400> 20
Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr
1 5 10 15
Asp Ala Arg Cys Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser
20 25 30
Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn
35 40 45
Val Arg Thr Asn Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
50 55 60
Lys Ala Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp
65 70 75 80
Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
85 90 95
Ser Leu Gln Pro Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Asn
100 105 110
Ser Tyr Pro Arg Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
115 120 125
Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr
145 150 155 160
Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
165 170 175
Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
180 185 190
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
195 200 205
His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
210 215 220
Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
225 230
<210> 21
<211> 445
<212> PRT
<213> artificial sequence
<220>
<223> mAb1 full heavy chain sequence (including Fc)
<400> 21
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45
Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr Tyr Asn Glu Lys Phe
50 55 60
Lys Gly Arg Ala Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr
65 70 75 80
Met Gln Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
85 90 95
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Ala Gln Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Gln Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 445
<210> 22
<211> 214
<212> PRT
<213> artificial sequence
<220>
<223> mAb1 full-length light chain sequence
<400> 22
Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
1 5 10 15
Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Thr Asn
20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile
35 40 45
Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly
50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
65 70 75 80
Glu Asp Phe Ala Asp Tyr Phe Cys Gln Gln Tyr Asn Ser Tyr Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala
100 105 110
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln
145 150 155 160
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205
Phe Asn Arg Gly Glu Cys
210
<210> 23
<211> 1392
<212> DNA
<213> artificial sequence
<220>
<223> DNA heavy chain sequence of anti-KIT antibody
<400> 23
atggagtggt cctgggtgtt cctgttcttt ctgtccgtga ccacaggcgt gcacagccag 60
gtgcagctgg tgcagtctgg agctgaggtg aagaagccag gagcttctgt gaagctgtcc 120
tgcaaggcca gcggctacac cttcacagac tactatatca actgggtgag acaggctcct 180
ggcaagggcc tggagtggat cgctcgcatc tatccaggct ctggcaacac ctactataat 240
gagaagttta agggccgggc caccctgaca gctgataaga gcacctctac agcttacatg 300
cagctgtcca gcctgagatc cgaggacacc gccgtgtact tctgcgctcg cggcgtgtac 360
tattttgatt attggggcca gggcaccaca gtgaccgtgt cttccgctag cacaaagggc 420
ccttccgtgt ttccactggc tcccagctct aagtccacca gcggaggaac agccgctctg 480
ggctgtctgg tgaaggacta tttcccagag cccgtgaccg tgagctggaa ctctggcgcc 540
ctgaccagcg gagtgcatac atttcctgct gtgctgcagt ccagcggcct gtactctctg 600
tcttccgtgg tgaccgtgcc aagctcttcc ctgggcaccc agacatatat ctgcaacgtg 660
aatcacaagc catccaatac aaaggtggac aagaaggtgg agcccaagag ctgtgataag 720
acccatacat gccccccttg tcctgctcca gaggctcagg gaggaccatc cgtgttcctg 780
tttccaccca agcctaagga caccctgtac atcacaaggg agccagaggt gacctgcgtg 840
gtggtggacg tgagccacga ggatcccgag gtgaagttca actggtacgt ggatggcgtg 900
gaggtgcata atgccaagac aaagccaagg gaggagcagt acaatagcac ctatcgggtg 960
gtgtctgtgc tgacagtgct gcaccaggac tggctgaacg gcaaggagta caagtgccag 1020
gtgtctaata aggccctgcc cgctcctatc gagaagacca tctccaaggc caagggccag 1080
cctagggagc cacaggtgta cacactgcct ccaagccggg acgagctgac caagaaccag 1140
gtgtctctga catgtctggt gaagggcttc tatccctctg atatcgctgt ggagtgggag 1200
tccaatggcc agcctgagaa caattacaag accacacccc ctgtgctgga ctccgatggc 1260
agcttctttc tgtattccaa gctgaccgtg gataagagca ggtggcagca gggcaacgtg 1320
ttttcttgtt ccgtgatgca tgaggctctg cacaatcatt acacacagaa gagcctgtct 1380
ctgtcccctg gc 1392
<210> 24
<211> 702
<212> DNA
<213> artificial sequence
<220>
<223> DNA light chain sequence of anti-KIT antibody
<400> 24
atgtccgtgc caacccaggt gctgggactg ctgctgctgt ggctgaccga cgccaggtgc 60
gatatcgtga tgacacagtc cccttccagc ctgtctgctt ccgtgggcga cagagtgacc 120
atcacctgta aggccagcca gaacgtgcgc accaatgtgg cttggtacca gcagaagcca 180
ggcaaggccc ccaaggctct gatctatagc gcctcttaca ggtatagcgg agtgcctgac 240
cggttcaccg gatccggaag cggaacagac ttcaccctga caatctcttc cctgcagcct 300
gaggacttcg ctgattactt ttgccagcag tacaactctt atccaaggac cttcggcggc 360
ggcacaaagg tggagatcaa gcggaccgtg gccgctccaa gcgtgttcat ctttccccct 420
tctgacgagc agctgaagtc tggcacagcc tccgtggtgt gcctgctgaa caacttctac 480
cccagagagg ccaaggtgca gtggaaggtg gataacgctc tgcagtctgg caattcccag 540
gagagcgtga ccgagcagga ctctaaggat tccacatata gcctgagctc taccctgaca 600
ctgtctaagg ccgattacga gaagcacaag gtgtatgctt gcgaggtgac ccatcagggc 660
ctgtccagcc cagtgacaaa gtccttcaat cgcggcgagt gt 702
<210> 25
<211> 7
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR1 of anti-KIT antibody (VH-CDR 1)
<400> 25
Gly Tyr Thr Phe Thr Asp Tyr
1 5
<210> 26
<211> 6
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR2 of anti-KIT antibody (VH-CDR 2)
<400> 26
Tyr Pro Gly Ser Gly Asn
1 5
<210> 27
<211> 8
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR3 of anti-KIT antibody (VH-CDR 3-AbM)
<400> 27
Gly Val Tyr Tyr Phe Asp Tyr Trp
1 5
<210> 28
<211> 7
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR1 of anti-KIT antibody (VL-CDR 1)
<400> 28
Ser Gln Asn Val Arg Thr Asn
1 5
<210> 29
<211> 3
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR2 of anti-KIT antibody (VL-CDR 2)
<400> 29
Ser Ala Ser
1
<210> 30
<211> 6
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR3 of anti-KIT antibody (VL-CDR 3)
<400> 30
Tyr Asn Ser Tyr Pro Arg
1 5
<210> 31
<211> 4
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR2 of anti-KIT antibody (VH-CDR 2)
<400> 31
Pro Gly Ser Gly
1
<210> 32
<211> 6
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR3 of anti-KIT antibody (VH-CDR 3)
<400> 32
Val Tyr Tyr Phe Asp Tyr
1 5
<210> 33
<211> 10
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR1 of anti-KIT antibody (VH-CDR 1-AbM)
<400> 33
Gly Tyr Thr Phe Thr Asp Tyr Tyr Ile Asn
1 5 10
<210> 34
<211> 10
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR2 of anti-KIT antibody (VH-CDR 2-AbM)
<400> 34
Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr
1 5 10
<210> 35
<211> 7
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR1 (VL-CDR 1-Contact) of an anti-KIT antibody
<400> 35
Arg Thr Asn Val Ala Trp Tyr
1 5
<210> 36
<211> 10
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR2 (VL-CDR 2-Contact) of an anti-KIT antibody
<400> 36
Ala Leu Ile Tyr Ser Ala Ser Tyr Arg Tyr
1 5 10
<210> 37
<211> 8
<212> PRT
<213> artificial sequence
<220>
<223> light chain CDR3 (VL-CDR 3-Contact) of an anti-KIT antibody
<400> 37
Gln Gln Tyr Asn Ser Tyr Pro Arg
1 5
<210> 38
<211> 6
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR1 (VH-CDR 1-Contact) of anti-KIT antibody
<400> 38
Thr Asp Tyr Tyr Ile Asn
1 5
<210> 39
<211> 13
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR2 (VH-CDR 2-Contact) of anti-KIT antibody
<400> 39
Trp Ile Ala Arg Ile Tyr Pro Gly Ser Gly Asn Thr Tyr
1 5 10
<210> 40
<211> 9
<212> PRT
<213> artificial sequence
<220>
<223> heavy chain CDR3 (VH-CDR 3-Contact) of anti-KIT antibody
<400> 40
Ala Arg Gly Val Tyr Tyr Phe Asp Tyr
1 5

Claims (32)

1. A method of protecting, treating, or controlling chronic prurigo in a subject comprising administering to a subject in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof that immunospecifically binds to human KIT.
2. The method of claim 1, wherein the human KIT comprises SEQ ID NO:1, and a polypeptide having the amino acid sequence shown in 1.
3. The method of claim 1 or 2, wherein the chronic prurigo is prurigo nodularis.
4. A method according to any one of claims 1 to 3, wherein the antibody is a bivalent monospecific antibody.
5. A method according to any one of claims 1 to 3, wherein the antibody is a bispecific antibody.
6. The method of any one of claims 1 to 5, wherein the antibody is a humanized antibody.
7. The method of any one of claims 1 to 6, wherein the antibody comprises a modified Fc region or domain.
8. The method of any one of claims 1-7, wherein the antibody has reduced Fc receptor binding activity.
9. The method of claim 8, wherein the antibody has reduced fcγr binding activity.
10. The method of any one of claims 1 to 9, wherein the antibody does not cause significant degranulation of FcgRI-expressing human mast cells.
11. The method of any one of claims 1-10, wherein the antibody does not exhibit significant Fc receptor-dependent KIT agonist activity.
12. The method of any one of claims 1 to 11, wherein the antibody specifically binds to the D4 or D5 region of human KIT.
13. The method of any one of claims 1 to 12, wherein the antibody comprises:
(A) (i) a light chain variable region ("VL") comprising a sequence comprising SEQ ID NO: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) A heavy chain variable region ("VH") comprising the amino acid sequences comprising SEQ ID NOs: 5. SEQ ID NO:6 and SEQ ID NO:7, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown;
(B) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:26 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no;
(C) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 28. SEQ ID NO:29 and SEQ ID NO:30, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 25. SEQ ID NO:31 and SEQ ID NO:32, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown;
(D) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 2. SEQ ID NO:3 and SEQ ID NO:4, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in fig; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 33. SEQ ID NO:34 and SEQ ID NO:27, VH CDR1, VH CDR2, and VH CDR3 of the amino acid sequence shown in seq id no; or alternatively
(E) (i) VL comprising amino acid sequences comprising SEQ ID NOs: 35. SEQ ID NO:36 and SEQ ID NO:37, VL CDR1, VL CDR2, and VL CDR3 of the amino acid sequence shown in seq id no; and
(ii) VH comprising amino acid sequences comprising SEQ ID NOs: 38. SEQ ID NO:39 and SEQ ID NO:40, VH CDR1, VH CDR2 and VH CDR3 of the amino acid sequence shown.
14. The method of claim 13, wherein the antibody comprises a VL and a VH, the VL comprising a sequence comprising SEQ ID NO:2-4, said VH comprising VL CDRs 1-3 comprising the amino acid sequences set forth in SEQ ID NOs: 5-7, and VH CDR 1-3 of the amino acid sequence shown.
15. The method of any one of claims 1 to 14, wherein the antibody comprises
(i) VL comprising the amino acid sequence:
DIVMTQSPSX K1 LSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKX K2 LIYSAS YRYSGVPDRFX K3 GSGSGTDFTLTISSLQX K4 EDFAX K5 YX K6 CQQYNSYPRTFGGGTKVE IK (SEQ ID NO: 17), wherein X K1 Is an amino acid having an aromatic or aliphatic hydroxyl side chain, X K2 Is an amino acid having an aliphatic or aliphatic hydroxyl side chain, X K3 Is an amino acid having an aliphatic hydroxyl side chain, X K4 Is an amino acid having an aliphatic hydroxyl side chain or P, X K5 Is an amino acid having a charged or acidic side chain and X K6 Is an amino acid having an aromatic side chain; and
(ii) VH comprising the amino acid sequence:
QVQLVQSGAEX H1 KKPGASVKX H2 SCKASGYTFTDYYINWVX H3 QAPGKGLEWIA RIYPGSGNTYYNEKFKGRX H4 TX H5 TAX H6 KSTSTAYMX H7 LSSLRSEDX H8 AVYFCARGV YYFDYWGQGTTVTVSS (SEQ ID NO: 18), wherein X H1 Is an amino acid having an aliphatic side chain, X H2 Is an amino acid having an aliphatic side chain, X H3 Is an amino acid having a polar or basic side chain, X H4 Is an amino acid having an aliphatic side chain, X H5 Is an amino acid having an aliphatic side chain, X H6 Is an amino acid with an acidic side chain, X H7 Is an amino acid having an acidic or amide derivative side chain and X H8 Is an amino acid having an aliphatic hydroxyl side chain.
16. The method of claim 15, wherein X K1 Is amino acid F or S, X K2 Is amino acid A or S, X K3 Is the amino acid T or S, X K4 Is the amino acid S or P, X K5 Is amino acid D or T, X K6 Is amino acid F or Y, X H1 Is amino acid L or V, X H2 Is amino acid L or V, X H3 Is amino acid K or R, X H4 Is the amino acid V or A, X H5 Is amino acid L or I, X H6 Is amino acid E or D, X H7 Is amino acid Q or E and X H8 Is amino acid S or T.
17. The method of any one of claims 1 to 16, wherein the antibody comprises a VL comprising an amino acid sequence selected from the group consisting of seq id nos: SEQ ID NO: 13. 14, 15 and 16; and VH comprising an amino acid sequence selected from the group consisting of: SEQ ID NO: 8. 9, 10, 11 and 12.
18. The method of any one of claims 1 to 17, wherein the antibody comprises a human light chain constant region.
19. The method of any one of claims 1 to 18, wherein the antibody comprises a human heavy chain constant region.
20. The method of claim 19, wherein the human heavy chain constant region is a human IgG constant region.
21. The method of claim 20, wherein the human heavy chain constant region is a human IgG1 constant region.
22. The method of any one of claims 1 to 21, wherein the antibody comprises a modified human Fc region or domain.
23. The method of any one of claims 1 to 21, wherein the antibody comprises a modified human IgG1 Fc region or domain.
24. The method of claim 23, wherein the modified human IgG1 Fc region or domain comprises non-naturally occurring amino acids 234A, 235Q, and 322Q as numbered by the EU index as set forth in Kabat.
25. The method of claim 24, wherein the modified human IgG1 Fc region or domain further comprises non-naturally occurring amino acids 252Y, 254T, and 256E as numbered by the EU index as set forth in Kabat.
26. The method of any one of claims 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q and 322Q as numbered by the EU index as set forth in Kabat.
27. The method of any one of claims 1 to 25, wherein the antibody comprises:
(i) Comprising SEQ ID NO:14, VL of the amino acid sequence shown in seq id no;
(ii) Comprising SEQ ID NO:10, VH of the amino acid sequence shown in fig; and
(iii) A modified human IgG1 Fc region or domain comprising non-naturally occurring amino acids 234A, 235Q, 322Q, 252Y, 254T and 256E as numbered by the EU index as set forth in Kabat.
28. The method of any one of claims 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO：21)。
29. the method of any one of claims 1 to 25, wherein the antibody comprises a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
30. the method of any one of claims 1 to 25, wherein the antibody comprises a heavy chain comprising the amino acid sequence:
QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYINWVRQAPGKGLEWIARIYPGSGNTYYNEKFKGRATLTADKSTSTAYMQLSSLRSEDTAVYFCARGVYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAQGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCQVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 21); and a light chain comprising the amino acid sequence:
DIVMTQSPSSLSASVGDRVTITCKASQNVRTNVAWYQQKPGKAPKALIYSASYRYSGVPDRFTGSGSGTDFTLTISSLQPEDFADYFCQQYNSYPRTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO：22)。
31. The method of any one of claims 1-30, wherein the subject is an adult.
32. The method of any one of claims 1 to 30, wherein the subject is a child.
CN202280023482.5A 2021-01-22 2022-01-21 Treatment of chronic prurigo Pending CN117120089A (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US63/140,621 2021-01-22
US202163238688P 2021-08-30 2021-08-30
US63/238,688 2021-08-30
PCT/US2022/013380 WO2022159744A1 (en) 2021-01-22 2022-01-21 Treatment of chronic prurigo

Publications (1)

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CN117120089A true CN117120089A (en) 2023-11-24

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Country Link
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