ANTI-CGRP COMPOSITIONS AND USE THEREOF
The present invention is directed to antibodies and fragments thereof having binding specificity for CGRP. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V H , V L and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-CGRP antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-CGRP antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-CGRP antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with CGRP.
1 . An anti-human CGRP antibody or antibody fragment which comprises the same VH and VL CDRs as an anti-human CGRP antibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13 or Ab14.
2 . (canceled)
3 . The antibody fragment of claim 1 , wherein
(i) said fragment is selected from a Fab fragment, a Fab′ fragment, a F(ab′)2 fragment, or a monovalent antibody such as “metMab”;
(ii) said fragment is a Fab fragment,
(iii) the anti-human CGRP antibody or antibody fragment comprises the same CDRs as an anti-human CGRP antibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13 or Ab14;
(iv) it comprises a variable light chain comprising the CDR 1 sequence of SEQ ID NO:25, the CDR 2 sequence of SEQ ID NO:26, and the CDR 3 sequence of SEQ ID NO:27, and/or a variable heavy chain comprising the CDR 1 sequence of SEQ ID NO:28, the CDR 2 sequence of SEQ ID NO:29, and the CDR 3 sequence of SEQ ID NO:30;
(v) it comprises a variable light chain comprising the CDR 1 sequence of SEQ ID NO:55, the CDR 2 sequence of SEQ ID NO:56, and the CDR 3 sequence of SEQ ID NO:57, and/or a variable heavy chain comprising the CDR 1 sequence of SEQ ID NO:58, the CDR 2 sequence of SEQ ID NO:59, and the CDR 3 sequence of SEQ ID NO:60;
(vi) it is aglycosylated or if glycosylated only contains only mannose residues;
(vii) it is not N-glycosylated;
(viii) it contains an Fc region that has been modified to alter effector function, half-life, proteolysis, and/or glycosylation;
(ix) it is a humanized, single chain or chimeric antibody; or
(x) it is directly or indirectly attached to a detectable label or therapeutic agent.
4 - 22 . (canceled)
23 . A nucleic acid sequence or nucleic acid sequences which encode an anti-human CGRP antibody or antibody fragment according to claim 1 , or a vector containing said nucleic acid sequence or nucleic acid sequences.
24 - 26 . (canceled)
27 . A cultured or recombinant cell, optionally a mammalian, yeast, bacterial, fungal, or insect cell, which expresses an antibody or antibody fragment according to claim 1 .
28 - 31 . (canceled)
32 . A method of treatment comprising administering to a patient with a disease or condition treatable by the administration of a CGRP antagonist a therapeutically effective amount of at least one anti-human CGRP antibody or fragment according to claim 1 .
33 . (canceled)
34 . The method of claim 32 wherein
(i) the disease is selected from migraine (with or without aura), weight loss, cancer or tumors, angiogenesis associated with cancer or tumor growth, angiogenesis associated with cancer or tumor survival, migraine, chronic migraine, frequent episodic migraines, or menstrual migraines hemiplagic migraines, cluster headaches, migrainous neuralgia, chronic headaches, tension headaches, general headaches, hot flushes, chronic paroxysomal hemicrania, secondary headaches due to an underlying structural problem in the head or neck, cranial neuralgia, sinus headaches (such as for example associated with sinusitis), allergy-induced headaches or migraines, pain, inflammatory pain, post-operative incision pain, complex regional pain syndrome, cancer pain, primary or metastatic bone cancer pain, fracture pain, osteoporotic fracture pain, pain resulting from burn, osteoporosis, gout joint pain, pain associated with sickle cell crises, and other nociceptic pain, as well as hepatocellular carcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathic pain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia, phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflex sympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoid arthritis pain, lower back pain, diabetic neuropathy, sciatica, or visceral pain associated with gastro-esophageal reflux, dyspepsia, irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis, menstrual period, labor, menopause, prostatitis, or pancreatitis;
(ii) the condition comprises overactive bladder; urinary incontinence; pain; chronic pain; neurogenic inflammation and inflammatory pain; neuropathic pain; eye pain; tooth pain; post-surgical pain, trauma related pain, diabetes; non-insulin dependent diabetes mellitus and other inflammatory autoimmune disorders, vascular disorders; inflammation; arthritis; bronchial hyperreactivity, asthma; shock; sepsis; opiate withdrawal syndrome; morphine tolerance; hot flashes in men and women; allergic dermatitis; psoriasis; encephalitis; brain trauma; epilepsy; neurodegenerative diseases; skin diseases including pruritis, neurogenic cutaneous redness, skin rosaceousness and erythema; inflammatory bowel disease, irritable bowel syndrome, cystitis; and dysmennorhea.
(iii) the disease or condition is pain, overactive bladder, urinary incontinence, headache or migraine;
(iv) the treatment further includes the administration of another therapeutic agent or regimen selected from anti-histamines, anti-inflammatory agents, analgesics or antibiotics;
(v) the treatment further includes the administration of an analgesic which comprises an NSAID, an opioid analgesic, an antibody or antibody fragment or another analgesic biologic;
(vi) the treatment further includes the administration of an analgesic which comprises an opioid;
(vii) the treatment further includes the administration of an analgesic which comprises morphine or a morphine derivative; or
(viii) the treatment further includes the administration of an analgesic which comprises an NGF antibody or antibody fragment.
35 - 44 . (canceled)
45 . An isolated anti-CGRP antibody or antibody fragment according to claim 1 , comprising a V H polypeptide sequence selected from: SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, or 133, or a variant thereof that exhibits at least 90% sequence identity therewith; and/or a V L polypeptide sequence selected from: SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121 or 131, or a variant thereof that exhibits at least 90% sequence identity therewith, wherein one or more of the framework (FR) in said V H or V L polypeptide has been substituted with another amino acid residue resulting in an anti-CGRP antibody that specifically binds CGRP.
46 - 60 . (canceled)
61 . The antibody or antibody fragment of claim 45 , wherein said antibody or antibody fragment further comprises an effector moiety, optionally a detectable moiety or a functional moiety, further optionally wherein said detectable moiety is a fluorescent dye, an enzyme, a substrate, a bioluminescent material, a radioactive material, or a chemiluminescent material and/or optionally said functional moiety is streptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, or a radioactive material.
62 - 64 . (canceled)
65 . A method of ameliorating or reducing symptoms of a disease or disorder characterized by pathological increased CGRP expression comprising administering to an individual in need thereof a therapeutically effective amount of a CGRP antibody or antibody fragment according to claim 1 .
66 . (canceled)
67 . The method of claim 65 , wherein
(i) said disease or disorder is selected from migraine (with or without aura), weight loss, cancer or tumors, angiogenesis associated with cancer or tumor growth, angiogenesis associated with cancer or tumor survival, hemiplagic migraines, cluster headaches, migrainous neuralgia, chronic headaches, tension headaches, general headaches, hot flushes, chronic paroxysomal hemicrania, secondary headaches due to an underlying structural problem in the head or neck, cranial neuralgia, sinus headaches (such as for example associated with sinusitis), allergy-induced headaches or migraines, pain, inflammatory pain, post-operative incision pain, complex regional pain syndrome, cancer pain, primary or metastatic bone cancer pain, fracture pain, osteoporotic fracture pain, pain resulting from burn, osteoporosis, gout joint pain, pain associated with sickle cell crises, and other nociceptic pain, as well as hepatocellular carcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathic pain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia, phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflex sympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoid arthritis pain, lower back pain, diabetic neuropathy, sciatica, or visceral pain associated with gastro-esophageal reflux, dyspepsia, irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis, menstrual period, labor, menopause, prostatitis, or pancreatitis;
(ii) the condition comprises overactive bladder, pain; TMJ, temporomandibular jaw disorders, chronic pain; neurogenic inflammation and inflammatory pain; neuropathic pain; eye pain; tooth pain; post-surgical pain, trauma related pain, diabetes; non-insulin dependent diabetes mellitus and other inflammatory autoimmune disorders, vascular disorders; inflammation; arthritis; sarcoidosis, bronchial hyperreactivity, asthma; shock; sepsis; opiate withdrawal syndrome; morphine tolerance; hot flashes in men and women; allergic dermatitis; psoriasis; encephalitis; brain trauma; epilepsy; neurodegenerative diseases; skin diseases including pruritis, neurogenic cutaneous redness, skin rosaceousness and erythema; inflammatory bowel disease, irritable bowel syndrome, cystitis; and dysmennorhea; or
(iii) the disease or condition is pain, overactive bladder, headache, or migraine.
68 - 81 . (canceled)
82 . A method of making the antibody or antibody fragment of claim 1 in a polyploid yeast culture that stably expresses and secretes into the culture medium at least 10 −25 mg/liter of said antibody, comprising:
(i) introducing at least one expression vector containing one or more heterologous polynucleotides encoding said antibody operably linked to a promoter and a signal sequence into a haploid yeast cell;
(ii) producing by mating or spheroplast fusion a polyploidal yeast from said first and/or second haploid yeast cell;
(iii) selecting polyploidal yeast cells that stably express said antibody; and
(iv) producing stable polyploidal yeast cultures from said polyploidal yeast cells that stably express said antibody into the culture medium.
83 - 85 . (canceled)
86 . An isolated polynucleotide comprising a polynucleotide encoding an anti-CGRP V H antibody amino acid sequence selected from SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123 or 133, or encoding a variant thereof which exhibits at least 90% sequence identity therewith and which variant comprises the same CDRs as the unmodified V H antibody amino acid sequence; and/or a polynucleotide sequence encoding an anti-CGRP V L amino acid sequence selected from SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121 or 131, or a polynucleotide encoding a variant thereof which exhibits at least 90% sequence identity therewith and which variant comprises the same CDRs as the unmodified V H antibody amino acid sequence.
87 - 93 . (canceled)
94 . A vector comprising the polynucleotide sequence of claim 86 .
95 . A host cell comprising the vector of claim 94 .
96 - 142 . (canceled)
143 . A pharmaceutical or diagnostic composition containing at least one CGRP antibody or fragment according to claim 1 and a pharmaceutically acceptable carrier.
144 - 187 . (canceled)
188 . An anti-CGRP antibody or antibody fragment according to claim 1 having binding specificity for CGRP and comprising variable light chain CDR1, CDR2, and CDR3 polypeptide sequences and variable heavy chain CDR1, CDR2, and CDR3 polypeptide sequences selected from the following:
V L
V L
V L
V H
V H
V H
CDR1
CDR2
CDR3
CDR1
CDR2
CDR3
A
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 5
No: 6
No: 7
No: 8
No: 9
No: 10
B
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 15
No: 16
No: 17
No: 18
No: 19
No: 20
C
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 25
No: 26
No: 27
No: 28
No: 29
No: 30
D
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 35
No: 36
No: 37
No: 38
No: 39
No: 40
E
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 45
No: 46
No: 47
No: 48
No: 49
No: 50
F
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 55
No: 56
No: 57
No: 58
No: 59
No: 60
G
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 65
No: 66
No: 67
No: 68
No: 69
No: 70
H
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 75
No: 76
No: 77
No: 78
No: 79
No: 80
I
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 85
No: 86
No: 87
No: 88
No: 89
No: 90
J
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 95
No: 96
No: 97
No: 98
No: 99
No: 100
K
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 105
No: 106
No: 107
No: 108
No: 109
No: 110
L
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 115
No: 116
No: 117
No: 118
No: 119
No: 120
M
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 125
No: 126
No: 127
No: 128
No: 129
No: 130
N
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
Seq ID
No: 135
No: 136
No: 137
No: 138
No: 139
No: 140
189 - 190 . (canceled)
191 . The anti-CGRP antibody or antibody fragment of claim 188 , comprising a variable light chain polypeptide sequence and a variable heavy chain polypeptide sequence selected from the following:
Variable Light
Variable Heavy
Chain
Chain
A
SEQ ID NO: 1
SEQ ID NO: 3
B
SEQ ID NO: 11
SEQ ID NO: 13
C
SEQ ID NO: 21
SEQ ID NO: 23
D
SEQ ID NO: 31
SEQ ID NO: 33
E
SEQ ID NO: 41
SEQ ID NO: 43
F
SEQ ID NO: 51
SEQ ID NO: 53
G
SEQ ID NO: 61
SEQ ID NO: 63
H
SEQ ID NO: 71
SEQ ID NO: 73
I
SEQ ID NO: 81
SEQ ID NO: 83
J
SEQ ID NO: 91
SEQ ID NO: 93
K
SEQ ID NO: 101
SEQ ID NO: 103
L
SEQ ID NO: 111
SEQ ID NO: 113
M
SEQ ID NO: 121
SEQ ID NO: 123
N
SEQ ID NO: 131
SEQ ID NO: 133
192 . The anti-CGRP antibody or antibody fragment of claim 188 , comprising a light chain polypeptide sequence and a heavy chain polypeptide sequence selected from the following:
Light Chain
Heavy Chain
Ab1
SEQ ID NO: 2
SEQ ID NO: 4
Ab2
SEQ ID NO: 12
SEQ ID NO: 14
Ab3
SEQ ID NO: 22
SEQ ID NO: 24
Ab4
SEQ ID NO: 32
SEQ ID NO: 34
Ab5
SEQ ID NO: 42
SEQ ID NO: 44
Ab6
SEQ ID NO: 52
SEQ ID NO: 54
Ab7
SEQ ID NO: 62
SEQ ID NO: 64
Ab8
SEQ ID NO: 72
SEQ ID NO: 74
Ab9
SEQ ID NO: 82
SEQ ID NO: 84
Ab10
SEQ ID NO: 92
SEQ ID NO: 94
Ab11
SEQ ID NO: 102
SEQ ID NO: 104
Ab12
SEQ ID NO: 112
SEQ ID NO: 114
Ab13
SEQ ID NO: 122
SEQ ID NO: 124
Ab14
SEQ ID NO: 132
SEQ ID NO: 134
193 - 211 . (canceled)