Treatment of the most bothersome migraine-related symptoms (MBS) with anti-CGRP antibodies

Abstract

Methods for treating migraine-associated most bothersome symptoms (MBS) are provided. Exemplary methods provide improvement in migraine-associated MBS within one month of administration of an anti-CGRP antibody of the invention. Methods for improving Patient Impression of Migraine-Associated Change (PGIC) are also provided. Exemplary methods include administering an anti-CGRP antagonist antibody to a patient in need thereof.

Description

[Technical field]

[0001] Disclosure of sequence listing This application contains a Sequence Listing that was submitted in ASCII format via EFS-Web and is incorporated herein by reference in its entirety. The ASCII copy was created on April 6, 2020, is named "1250-US-PSP_ST25.txt" and is 349KB in size.

[0002] FIELD OF THEINVENTION The present invention relates to a method for treating the most bothersome symptoms associated with migraine using an antibody or a fragment thereof (including a Fab fragment) that specifically binds to human calcitonin gene-related peptide (hereinafter referred to as "CGRP"). [Background technology]

[0003] 2. Description of Related Art Calcitonin gene-related peptide (CGRP) is produced as a 37 amino acid long multifunctional neuropeptide. Two forms of CGRP, CGRP-α and CGRP-β forms, exist in humans and have similar activity. CGRP-α and CGRP-β differ in humans by three amino acids and originate from different genes. CGRP is released from numerous tissues, including the trigeminal nerve, and upon activation, neuropeptides are released in the meninges to mediate neurogenic inflammation characterized by vasodilation, vascular leakage, and mast cell degradation. Durham, PL, New Eng. J. Med., 350(11):1073-75 (2004). The biological actions of CGRP are mediated through the seven-transmembrane CGRP receptor (CGRP-R) by binding to receptor-associated membrane proteins (RAMPs). The CGRP-R further requires the activity of receptor component proteins (RCPs), which are essential for efficient G protein-mediated coupling to adenylate cyclase and the production of cAMP. Doods, H., Curr. Op. Invest. Drugs, 2(9):1261-68 (2001).

[0004] Migraine is a neurovascular disorder that affects about 10% of the adult population in the United States and is typically accompanied by severe headaches. CGRP is believed to play a prominent role in the pathogenesis of migraine. Indeed, several companies, namely Amgen, Eli Lilly, Teva and Alder Biopharmaceuticals (recently acquired by Lundbeck A / S), have developed anti-CGRP and anti-CGRP-R antibodies for use in the treatment or prevention of migraine. The Assignee is the subject of PCT Publication No. WO 2012 / 162243, filed May 21, 2012, entitled "ANTI-CGRP COMPOSITIONS AND USE THEREOF", and PCT Publication No. WO 2012 / 162257, filed May 21, 2012, entitled "USE OF ANTI-CGRP ANTIBODIES AND ANTIBODY FRAGMENTS TO PREVENT OR INHIBIT PHOTOPHOBIA OR LIGHT AVERSION IN SUBJECTS IN NEED THEREOF, ESPECIALLY MIGRAINE SUFFERERS", and is hereby incorporated by reference in its entirety. The company has previously filed patent applications relating to anti-CGRP antibodies and uses thereof, including PCT Application Publication No. WO 2012 / 162253, filed May 21, 2012, entitled "REGULATION OF GLUCOSE METABOLISM USING ANTI-CGRP ANTIBODIES," and PCT Application Publication No. WO 2015 / 003122, filed July 3, 2014, entitled "REGULATION OF GLUCOSE METABOLISM USING ANTI-CGRP ANTIBODIES," all of which applications are incorporated by reference in their entireties. Summary of the Invention

[0005] The present disclosure provides a method for treating migraine-related most bothersome symptoms (MBS) in a patient suffering from chronic migraine, comprising administering to a patient in need thereof an effective amount of at least one anti-CGRP antibody or antibody fragment thereof, or anti-CGRP-R antibody or antibody fragment thereof, or one or more formulations comprising said antibody or antibody fragment disclosed herein. The antibody treatment may be initiated in the interictal period, i.e., between migraine attacks, or in the ictal period, i.e., during a migraine episode. The migraine may include, for example, chronic migraine or episodic migraine, in certain embodiments of the present invention where the patient suffers from chronic migraine. In the present invention, the anti-CGRP antibody or antibody fragment is designated Ab6. Ab6 is an anti-CGRP antibody or antibody fragment thereof having light chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, respectively; or having light chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively. The anti-CGRP antibody may comprise a variable light chain polypeptide of SEQ ID NO:222 and a variable heavy chain polypeptide of SEQ ID NO:202. The anti-CGRP antibody may comprise a variable light chain polypeptide encoded by SEQ ID NO:232 and a variable heavy chain polypeptide encoded by SEQ ID NO:212. The anti-CGRP antibody may comprise a light chain polypeptide of SEQ ID NO:221 and a heavy chain polypeptide of SEQ ID NO:201 or SEQ ID NO:566. The anti-CGRP antibody may comprise a light chain polypeptide encoded by SEQ ID NO:231 and a heavy chain polypeptide encoded by SEQ ID NO:211 or SEQ ID NO:567.The anti-CGRP antibody may comprise an antibody expression product isolated from a recombinant cell expressing a nucleic acid sequence encoding the variable light chain polypeptide of SEQ ID NO:222 and the variable heavy chain polypeptide of SEQ ID NO:202, which are optionally linked to human light and heavy chain constant region polypeptides, e.g., human IgG1, IgG2, IgG3 or IgG4 constant regions, respectively, which may optionally be modified to alter glycosylation or proteolysis, and the recombinant cell optionally comprises a yeast or mammalian cell, e.g., a Pichia pastoris or a CHO cell. The anti-CGRP antibody can include an antibody expression product isolated from a recombinant cell expressing a nucleic acid sequence encoding a light chain polypeptide of SEQ ID NO:221 and a heavy chain polypeptide of SEQ ID NO:201 or SEQ ID NO:566, the recombinant cell optionally including a yeast or mammalian cell, e.g., Pichia pastoris or CHO cell, the constant region of which can optionally be modified to alter glycosylation or proteolysis or other effector function. Any of the above-mentioned anti-CGRP antibodies or antibody fragments, preferably Ab6, can optionally be included in a formulation disclosed herein, e.g., histidine (L-histidine), sorbitol, polysorbate 80, e.g., about 100 mg of anti-CGRP antibody, about 3.1 mg of L-histidine, about 40.5 mg of sorbitol, and about 0.15 mg of polysorbate 80, having a pH of about 5.8, per mL volume. The dosage of the antibody may be from about 100 mg to about 300 mg, for example, about 100 mg, about 300 mg, 100 mg, or 300 mg. Doses may be administered by different means, for example, intravenously, for example, in an appropriate volume (e.g., 100 mL) of saline, such as 0.9% sodium chloride.

[0006] The patient may have less than 25 headache days per month, less than 20 headache days per month, less than 15 headache days per month, or less than 10 headache days per month. For example, the patient may have less than 14 headache days, less than 13 headache days, less than 12 headache days, less than 11 headache days, less than 10 headache days, less than 9 headache days, less than 8 headache days, less than 7 headache days, or less than 6 headache days per month. The patient may have 2-15 headache days per month, e.g., 3-14 headache days, 4-13 headache days, 5-12 headache days, 6-11 headache days, or 7-10 headache days.

[0007] The patient may have less than 10 migraines per month, such as 1-9 migraines per month, such as 2-8 migraines per month, 3-7 migraines per month, 4-6 migraines per month, or about 5 migraines per month. The patient may have less than 1 migraine per month, on average, such as once every 2 months, once every 3 months, once every 4 or 6 months, or a median number of migraines, such as twice every 3 months. The migraine may be diagnosed according to ICHD-3 guidelines.

[0008] In addition to the headache and associated symptoms described in the International Classification of Headache Disorders (ICHD-3) diagnostic criteria for migraine with or without aura, migraine patients experience a variety of autonomic, cognitive, sensory and motor symptoms during a migraine attack, which are experienced uniquely by each individual patient. In the present invention, patients were allowed to self-identify the specific symptoms associated with chronic migraine that they find most bothersome. In the present application, these symptoms will be referred to as the most bothersome symptoms (MBS) associated with migraine. In the present invention, patients are allowed to identify their own MBS without restriction, which provides a unique patient-centered approach to identify and measure the efficacy of the antibodies of the present invention as a treatment for these most bothersome symptoms associated with migraine, and is therefore expected to have a meaningful impact on the patient's ability to function during a migraine attack. Nausea, vomiting, photophobia, and phonophobia are symptoms related to migraine that are included in the ICDH-3 diagnostic criteria, but many other symptoms can be observed to occur before, after, and even during days with diagnosable migraine.During migraine attacks, these can include cognitive symptoms (e.g., memory impairment, executive dysfunction, attention disorder), emotional symptoms (e.g., mood swings, depression, anxiety, irritability), other sensory symptoms (e.g., odour phobia, taste disorder), as well as blurred vision, nasal congestion, rhinorrhea, lacrimation, sweating, ptosis, yawning, polyuria, abdominal cramps, diarrhea, dizziness, and neck pain.The migraine-related MBS reported by patients enrolled in the clinical trial described in Example 2 is summarized in Table 1. Although nausea / vomiting, photophobia, and phonophobia were common in the patient population in Example 2, fewer than half of these patients listed one of these three symptoms included in the ICDH-3 diagnostic criteria as their patient-identified MBS.

[0009] Migraine is a complex disorder of the brain associated with a multifaceted symptomatology that also manifests in a separate and unique way. The migraine pre- and post-ictal phases often last for many days and can be divided into four different phases - prodromal / prodromal, preictal / aura, attack / headache, postictal / postictal - with overlapping symptoms occurring during each migraine phase. The various types and timing of MBS during the course of a migraine are illustrated in Figure 15. It is highly appropriate to assess MBS in migraine patients during clinical trials, as it is recognized that headache alone is not considered sufficient to adequately relieve the impact of migraine on a patient's daily life and well-being. Reduction in mean monthly migraine days (MMD) or similar endpoints in clinical trials does not adequately capture the burden of migraine and associated symptoms that are affected by therapeutic intervention. The inventors of the present invention have discovered that in addition to reducing MMD, Ab6 (anti-CGRP antibody) is also effective in improving MBS in migraine patients. These treatment-related symptom improvements correlated with improvements in the patient's perception of their disease state and indirectly correlated with satisfaction with the treatment response. It is known that migraine patients often continue to seek treatment for their migraines due to the burden of MBS, thus supporting the clinical value of treating both the primary migraine pathology and the MBS associated with said migraines.

[0010] The present invention provides anti-CGRP antibodies or antibody fragments thereof that can improve MBS associated with migraine in patients suffering from migraine, such as chronic migraine or episodic migraine. The MBS parameter assesses the patient's assessment of the change in their symptoms (improvement or worsening since the start of the study).

[0011] The present invention provides an anti-CGRP antibody or antibody fragment thereof capable of improving the patient global impression of change associated with migraine treatment (PGIC) in patients suffering from migraine, such as chronic or episodic migraine. The patient global impression of change associated with migraine (PGIC) parameter comprises a single question regarding the patient's own impression of the overall change in the patient's disease status (improvement or worsening since the start of the study), which is rated on a 7-point Likert scale with the axes being very much improved and very much worse.

[0012] The present invention provides anti-CGRP antibodies or antibody fragments thereof that can reduce MMD and improve a patient's migraine-associated most bothersome symptoms (MBS) in a manner that is highly correlated with favorable changes in Patient Global Impression of Change (PGIC) of migraine treatment. This dual action represents an improved treatment option for patients suffering from migraine over treating migraine, providing treatment for the overall migraine burden experienced by the patient, including both migraine and migraine-associated MBS.

[0013] The present invention provides a method of ameliorating the most bothersome symptoms associated with migraine (MBS), the method comprising intravenously administering to a patient in need thereof about 100 mg to about 300 mg of an anti-CGRP antibody, the anti-CGRP antibody preferably comprising a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or 566.

[0014] The invention provides a method of improving Patient Global Impression of Change (PGIC), the method comprising intravenously administering to a patient in need thereof about 100 mg to about 300 mg of an anti-CGRP antibody, wherein the anti-CGRP antibody preferably comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or 566.

[0015] In another aspect, the invention provides a method of ameliorating the most bothersome symptoms associated with migraine (MBS) and simultaneously reducing MMD, the method comprising intravenously administering to a patient in need thereof about 100 mg to about 300 mg of an anti-CGRP antibody, wherein the anti-CGRP antibody preferably comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or 566.

[0016] In another aspect, the invention provides a method of improving Patient Global Impression of Changes (PGIC) associated with migraine and concomitantly reducing MMD, the method comprising intravenously administering to a patient in need thereof about 100 mg to about 300 mg of an anti-CGRP antibody, the anti-CGRP antibody preferably comprising a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or 566.

[0017] In another aspect, the invention provides a method of improving migraine associated Most Bothersome Symptoms (MBS) and Patient Global Impression of Changes (PGIC) associated with migraine, the method comprising intravenously administering to a patient in need thereof about 100 mg to about 300 mg of an anti-CGRP antibody, wherein the anti-CGRP antibody preferably comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or 566.

[0018] In another aspect, the invention provides a method of improving migraine-related Most Bothersome Symptoms (MBS) and / or Patient Global Impression of Migraine-Related Changes (PGIC) while simultaneously reducing MMD, the method comprising intravenously administering to a patient in need thereof about 100 mg to about 300 mg of an anti-CGRP antibody, wherein the anti-CGRP antibody preferably comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or 566.

[0019] In some exemplary embodiments, the dosage of the anti-CGRP antibody may be 100 mg.

[0020] In another exemplary embodiment, the dose of the anti-CGRP antibody may be 300 mg.

[0021] The method may further comprise intravenously administering 100 mg of the anti-CGRP antibody every 10 to 14 weeks, preferably every 11 to 13 weeks, and more preferably every 12 weeks.

[0022] The method may further comprise intravenously administering 300 mg of the anti-CGRP antibody every 10 to 14 weeks, preferably every 11 to 13 weeks, and more preferably every 12 weeks.

[0023] The antibody may be provided or administered in a formulation disclosed herein, e.g., comprising histidine (L-histidine), sorbitol, polysorbate 80, e.g., about 100 mg of anti-CGRP antibody, about 3.1 mg L-histidine, about 40.5 mg sorbitol, and about 0.15 mg polysorbate 80 per mL volume, having a pH of about 5.8.

[0024] Prior to the first dose, the patient may experience from about 10 to about 22 migraine days per month, such as from about 13 to about 19 migraine days per month, such as about 16 migraine days per month.

[0025] Prior to the first dose, the patient may experience about 14 to about 27 headache days per month, such as about 17 to about 24 headache days per month, such as about 20 or about 21 headache days per month.

[0026] The patient may have been diagnosed with migraine at least 10 years prior to the first dose, such as at least 15 years prior to the first dose, such as at least 18 years or at least 19 years prior to the first dose.

[0027] The patient may have been diagnosed with chronic migraine headaches at least 5 years prior to the first dose, such as at least 8 years prior to the first dose, such as at least 11 years or at least 12 years prior to the first dose.

[0028] The patient may have a headache upon administration of the anti-CGRP antibody, or fragment thereof, of the invention.

[0029] The patient may have a migraine headache, e.g., a migraine headache with aura, at the time of administration of the anti-CGRP antibody, or fragment thereof, of the invention.

[0030] The patient may have at least a 50% reduction in the number of migraine days during the month after administration of the first dose compared to the baseline number of migraine days experienced by the patient prior to the first dose.

[0031] The patient may have at least a 75% reduction in the number of migraine days during the month after administration of the first dose compared to the baseline number of migraine days experienced by the patient prior to the first dose.

[0032] The patient may have a 100% reduction in the number of migraine days during the month after administration of the first dose compared to the baseline number of migraine days experienced by the patient prior to the first dose.

[0033] The patient may have at least a 50% reduction in the number of migraine days in the 12 weeks after administration of the first dose compared to the baseline number of migraine days experienced by the patient prior to the first dose.

[0034] The patient may have at least a 75% reduction in the number of migraine days during the 12 weeks after administration of the first dose compared to the baseline number of migraine days experienced by the patient prior to the first dose.

[0035] The patient may have a 100% reduction in migraine days in the 12 weeks after administration of the first dose compared to the baseline number of migraine days experienced by the patient prior to the first dose.

[0036] The patient may experience improvement in migraine-associated MBS, measured as a change from baseline MBS, one month after administration of the first dose.

[0037] The patient may experience improvement in migraine-associated MBS, measured as a change from baseline MBS, over a three-month period following administration of the first dose.

[0038] The patient may experience improvement in migraine-associated MBS, measured as a change from baseline MBS, six months after administration of the first dose.

[0039] The patient may experience improvement in migraine-related PGIC, measured as a change from baseline, one month after administration of the first dose.

[0040] The patient may experience improvement in migraine-related PGIC, measured as a change from baseline, three months after administration of the first dose.

[0041] The patient may experience improvement in migraine-related PGIC, measured as a change from baseline, six months after administration of the first dose.

[0042] The method may further comprise administering to the patient, for example intravenously, a second dose of an anti-CGRP antibody of the invention within about 10 to 14 weeks, preferably 11 to 13 weeks, and more preferably about 12 weeks or about 3 months, after the first dose.

[0043] The first dose can comprise about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg of the anti-CGRP antibody.

[0044] Prior to said administration, the patient may exhibit about 15 to about 30 migraine days per month, such as about 16 to about 28 migraine days per month, such as about 17 to about 26 migraine days per month, such as about 16 migraine days per month.

[0045] Prior to said administration, the patient may experience about 15 to about 27 headache days per month, such as about 17 to about 24 headache days per month, such as about 20 or about 21 headache days per month.

[0046] The patient may have been diagnosed with migraine at least 10 years prior to said administering, such as at least 15 years prior to said administering, such as at least 18 years or at least 19 years prior to said administering.

[0047] The patient may have been diagnosed with chronic migraine headaches at least 5 years prior to said administering, such as at least 8 years prior to said administering, such as at least 11 years or at least 12 years prior to said administering.

[0048] The patient may have at least a 50% reduction in the number of migraine days during the month after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[0049] The patient may have at least a 75% reduction in the number of migraine days during the month after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[0050] The patient may have a 100% reduction in the number of migraine days over a one-month period after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[0051] The patient may have at least a 50% reduction in the number of migraine days over a 12 week period after administration of the antibody, compared to a baseline number of migraine days experienced by the patient prior to said administration.

[0052] The patient may have at least a 75% reduction in the number of migraine days over a 12 week period after administration of the antibody, compared to a baseline number of migraine days experienced by the patient prior to said administration.

[0053] The patient may have a 100% reduction in the number of migraine days over a 12 week period after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[0054] The method may further include administering a second dose of the anti-CGRP antibody to the patient, e.g., intravenously, within about 10 to 14 weeks, preferably 11 to 13 weeks, and more preferably about 12 weeks or about 3 months, after the administration.

[0055] The administration may comprise about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg of the anti-CGRP antibody.

[0056] The anti-CGRP antibody may be aglycosylated, or, if glycosylated, may contain only mannose residues.

[0057] The anti-CGRP antibody may consist of a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566. The anti-CGRP antibody may consist of a light chain polypeptide encoded by SEQ ID NO: 231 and a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0058] In some embodiments, the anti-human CGRP antibody or antibody fragment comprises a variable light chain of SEQ ID NO: 222 and / or a variable heavy chain of SEQ ID NO: 202. In some embodiments, the anti-human CGRP antibody or antibody fragment comprises a variable light chain encoded by SEQ ID NO: 232 and / or a variable heavy chain encoded by SEQ ID NO: 212.

[0059] In some embodiments, the anti-human CGRP antibody or antibody fragment comprises a light chain of SEQ ID NO: 221 and / or a heavy chain of SEQ ID NO: 201 or SEQ ID NO: 566. In some embodiments, the anti-human CGRP antibody or antibody fragment comprises a light chain encoded by SEQ ID NO: 231 and / or a heavy chain encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0060] In some embodiments, the anti-CGRP antibody comprises the V L Polypeptide and V of SEQ ID NO: 202 H The antibody may include antibody expression products isolated from recombinant cells expressing nucleic acid sequences encoding polypeptides, which are optionally linked to human light and heavy chain constant region polypeptides, e.g., human IgG1, IgG2, IgG3 or IgG4 constant regions, respectively, which may optionally be modified to alter glycosylation or proteolysis, and the recombinant cells optionally include yeast or mammalian cells, e.g., Pichia pastoris or CHO cells.

[0061] In some embodiments, the anti-CGRP antibody can include an antibody expression product isolated from a recombinant cell expressing a nucleic acid sequence encoding a light chain polypeptide of SEQ ID NO:221 and a heavy chain polypeptide of SEQ ID NO:201 or SEQ ID NO:566, the recombinant cell optionally comprising a yeast or mammalian cell, e.g., a Pichia pastoris or a CHO cell, the constant region of which can optionally be modified to alter glycosylation or proteolysis or other effector function.

[0062] In some embodiments, any of the foregoing anti-CGRP antibodies or antibody fragments may be included in a formulation disclosed herein, for example, having a pH of about 5.8, that includes histidine (L-histidine), sorbitol, polysorbate 80, for example, about 100 mg of anti-CGRP antibody, about 3.1 mg of L-histidine, about 40.5 mg of sorbitol, and about 0.15 mg of polysorbate 80 per mL volume. The antibody or fragment may be administered by different means, for example, intravenously, for example, in an appropriate volume (e.g., 100 mL) of saline, such as 0.9% sodium chloride.

[0063] In some embodiments, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg of the anti-CGRP antibody or antibody fragment is administered, e.g., intravenously.

[0064] In another embodiment, about 100 mg of the anti-CGRP antibody or antibody fragment is administered.

[0065] In another embodiment, about 300 mg of the anti-CGRP antibody or antibody fragment is administered, e.g., intravenously.

[0066] In exemplary embodiments, the anti-human CGRP antibody or antibody fragment is administered, e.g., intravenously, at most every 10-14 weeks, preferably every 11-13 weeks, more preferably every 3 months or every 12 weeks, and the antibody dosage is administered in a single formulation or split into different formulations that are administered about every 10-14 weeks, preferably every 11-13 weeks, more preferably every 3 months or every 12 weeks. The phrase "antibody dosage is administered in a single formulation or split into different formulations" refers to administration of the recited amount of antibody within a relatively short period of time, e.g., a few hours, e.g., 1-8 hours, within about 1 day, within about 2 days, or within about 1 week, which may be by the same or different routes of administration (e.g., intravenous, im, and / or sc). The term "different formulations" in this context refers to antibody doses that are administered at different times and / or at different sites and / or by different routes, regardless of whether the doses are the same or different in terms of the chemical composition of the pharmaceutical formulation in which each dose is administered; e.g., concentrations, excipients, carriers, pH, etc. may be the same or different between the different doses.

[0067] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 8 weeks or every two months.

[0068] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered about every 10-14 weeks, preferably every 11-13 weeks, and more preferably every 12 weeks or every 3 months.

[0069] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 16 weeks or every 4 months.

[0070] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 20 weeks or every 5 months.

[0071] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 24 weeks or every 6 months.

[0072] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 28 weeks or every 7 months.

[0073] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 32 weeks or every 8 months.

[0074] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 36 weeks or every 9 months.

[0075] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 40 weeks or every 8 months.

[0076] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 44 weeks or every 9 months.

[0077] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 48 weeks or every 10 months.

[0078] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 52 weeks or every 11 months.

[0079] In other exemplary embodiments, the dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations that are administered at a frequency of about every 56 weeks or every 12 months.

[0080] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations, which are administered at a frequency of about every 15-18 months.

[0081] In other exemplary embodiments, a dose of anti-human CGRP antibody or antibody fragment is administered in a single formulation or split into different formulations, which are administered at a frequency of about every 18-21 months.

[0082] In other exemplary embodiments, the anti-human CGRP antibody dose or antibody fragment used in the above methods is administered in a single formulation or is divided into different formulations that are administered at a frequency of about every two years.

[0083] In another exemplary embodiment, the anti-human CGRP antibody used in the above method is administered systemically.

[0084] In other exemplary embodiments, the anti-human CGRP antibody or antibody fragment used in the aforementioned methods is administered by a mode of administration selected from intravenous, intramuscular, intravenous, intrathecal, intracranial, topical, intranasal, and oral. In a preferred embodiment, the anti-human CGRP antibody or antibody fragment used in the aforementioned methods is administered intravenously.

[0085] In other exemplary embodiments, the anti-human CGRP antibody used in the above methods has an in vivo half-life of at least 10 days.

[0086] In other exemplary embodiments, the anti-human CGRP antibody has an in vivo half-life of at least 15 days.

[0087] In other exemplary embodiments, the anti-human CGRP antibody used in the above methods has an in vivo half-life of at least 20 days.

[0088] In other exemplary embodiments, the anti-human CGRP antibody used in the aforementioned methods has an in vivo half-life of at least 20-30 days.

[0089] In other exemplary embodiments, the anti-human CGRP antibody is administered at a dosage of about 100 mg to about 300 mg and has an in vivo half-life of at least about (284±44 hours)±20%.

[0090] In another exemplary embodiment, the anti-human CGRP antibody used in the above method binds to human α- and β-CGRP.

[0091] In another exemplary embodiment, the administered anti-human CGRP antibody dosage results in inhibition of vasodilation induced by locally applied capsaicin at least 30 days after antibody administration.

[0092] In another exemplary embodiment, the administered anti-human CGRP antibody dosage results in inhibition of vasodilation induced by topically applied capsaicin at least 60 days after antibody administration.

[0093] In another exemplary embodiment, the administered anti-human CGRP antibody dosage results in inhibition of vasodilation induced by locally applied capsaicin at least 90 days after administration of the antibody.

[0094] In another exemplary embodiment, the administered anti-human CGRP antibody dosage results in inhibition of vasodilation induced by locally applied capsaicin at least 120 days after antibody administration.

[0095] In another exemplary embodiment, the administered anti-human CGRP antibody dosage results in inhibition of vasodilation induced by locally applied capsaicin at least 150 days after administration of the antibody.

[0096] In another exemplary embodiment, the administered anti-human CGRP antibody dosage results in inhibition of vasodilation induced by locally applied capsaicin at least 180 days after administration of the antibody.

[0097] In another exemplary embodiment, the administered anti-human CGRP antibody dose results in inhibition of vasodilation induced by locally applied capsaicin more than 180 days after antibody administration.

[0098] In other exemplary embodiments, the administered anti-human CGRP antibody dose provides sustained pharmacodynamic (PK) activity within 5% of the maximum response (Imax) (compared to lower antibody doses).

[0099] In other exemplary embodiments, the administered anti-human CGRP antibody dose results in sustained pharmacodynamic (PK) activity that is maintained for at least 2-3 months after antibody administration, and where PK analysis of the anti-human CGRP antibody is derived from plasma concentrations.

[0100] In other exemplary embodiments, the administered anti-human CGRP antibody dosage is from about 100 mg to about 300 mg or more, which is administered no more frequently than every two months.

[0101] The present invention further relates to the use of specific antibodies and fragments thereof with binding specificity for CGRP, particularly antibodies with desired epitope specificity, high affinity or avidity and / or functional properties. A preferred embodiment of the present invention relates to the use of chimeric or humanized antibodies and fragments thereof (including Fab fragments) capable of binding to CGRP and / or inhibiting biological activity mediated by the binding of CGRP to the CGRP receptor ("CGRP-R"), for example such antibodies, optionally derived from recombinant cells engineered to express them, optionally yeast or mammalian cells, and further optionally Pichia pastoris and CHO cells.

[0102] In another preferred embodiment of the invention, full length and Fab fragments thereof are contemplated to inhibit CGRP-α-, CGRP-β-, and rat CGRP-driven production of cAMP. In a further preferred embodiment of the invention, full length and Fab fragments thereof are contemplated to reduce vasodilation in a recipient following administration.

[0103] The present invention also contemplates the use of conjugates of anti-CGRP antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The present invention also contemplates the use of chimeric or humanized anti-CGRP or anti-CGRP / CGRP-R complex antibodies and binding fragments thereof. In one embodiment, the binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, scFv fragments, SMIPs (small molecule immunopharmaceuticals), camelbodies, nanobodies, and IgNARs. [Brief description of the drawings]

[0104] [Figure 1] The polypeptide sequence of the full-length heavy chain of antibody Ab6 is provided, with the framework regions (FR), complementarity determining regions (CDR), and constant region sequences delineated. [Diagram 2] The polypeptide sequence of the full-length light chain of antibody Ab6 is provided, with the framework regions (FR), complementarity determining regions (CDR), and constant region sequences delineated. [Figure 3-1] An exemplary polynucleotide sequence encoding the full-length heavy chain of antibody Ab6 is provided, with the framework regions (FR), complementarity determining regions (CDR), and variable region coding sequences separated. [Figure 3-2] Same as description for Figure 3-1. [Figure 4] The polynucleotide sequence of the full-length light chain of antibody Ab6 is provided, clearly indicating its framework regions (FR), complementarity determining regions (CDR), and variable region coding sequences. [Diagram 5]1 provides the polypeptide sequence coordinates within the full-length heavy chain polypeptide sequence of antibody Ab6 of sequence features, including variable regions and complementarity determining regions (CDRs), as well as the SEQ ID NO of each of the individual features. [Figure 6] 1 provides the polypeptide sequence coordinates within the full-length heavy chain polypeptide sequence of antibody Ab6 of sequence features, including framework regions (FR) and constant regions, as well as the SEQ ID NO of each of the individual features. [Figure 7] 1 provides polypeptide sequence coordinates within the full-length light chain polypeptide sequence of antibody Ab6 of sequence features, including variable regions and complementarity determining regions (CDRs), as well as the SEQ ID NO of each of the individual features. [Figure 8] 1 provides the polypeptide sequence coordinates within the full-length light chain polypeptide sequence of antibody Ab6 of sequence features, including framework regions (FR) and constant regions, as well as the SEQ ID NO of each of the individual features. [Figure 9] Polynucleotide sequence coordinates within an exemplary polynucleotide sequence encoding the full-length heavy chain polypeptide sequence of antibody Ab6 of sequence features, including variable regions and complementarity determining regions (CDRs), are provided, as well as the SEQ ID NO of each of the individual features. [Figure 10] Polynucleotide sequence coordinates within an exemplary polynucleotide sequence encoding the full-length heavy chain polypeptide sequence of antibody Ab6 of sequence features, including framework regions (FR) and constant regions, are provided, as well as the SEQ ID NO of each of the individual features. [Figure 11] Polynucleotide sequence coordinates within an exemplary polynucleotide sequence encoding the full-length light chain polypeptide sequence of antibody Ab6 of sequence features, including variable regions and complementarity determining regions (CDRs), are provided, as well as the SEQ ID NO of each of the individual features. [Figure 12] Polynucleotide sequence coordinates within an exemplary polynucleotide sequence encoding the full-length light chain polypeptide sequence of antibody Ab6 of sequence features, including framework regions (FR) and constant regions, are provided, as well as the SEQ ID NO of each of the individual features. [Figure 13] Study design of the clinical trial protocol summarized in Example 2. [Figure 14]FIG. 1 shows the efficacy of Ab6 on mean monthly migraine days (MMD) in the clinical trial described in Example 2. [Figure 15] The type and timing of most bothersome symptoms (MBS) during the course of a migraine headache are indicated. [Figure 16] 1 shows the change in MBS from baseline during the 28-day screening period of the clinical trial described in Example 2 - i.e., prior to the first infusion of Ab6. [Figure 17] 1 shows the change in MBS from baseline one month after the first infusion of Ab6 in the clinical trial described in Example 2. [Figure 18] 1 shows PGIC from baseline one month after the first infusion of Ab6 in the clinical trial described in Example 2. [Figure 19] 1 shows the change from baseline in MBS 3 months after the first infusion of Ab6 in the clinical trial described in Example 2. [Figure 20] 1 shows PGIC from baseline 3 months after the first infusion of Ab6 in the clinical trial described in Example 2. [Figure 21] 1 shows the change from baseline in MBS 6 months after the first infusion of Ab6 in the clinical trial described in Example 2. [Figure 22] 1 shows PGIC from baseline 6 months after the first infusion of Ab6 in the clinical trial described in Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0105] Detailed Description The use of anti-CGRP antibody for the treatment of MBS and / or PGIC associated with migraine, for example chronic migraine or episodic migraine, is described herein.In addition, anti-CGRP antibody is demonstrated herein to be effective in the treatment of MMD.The treatment efficacy for both MBS and PGIC is shown to be effective in providing relief of MBS and PGIC at 1 month, 3 months and 6 months after the first injection of anti-CGRP antibody or its fragment of the present invention.

[0106] definition It should be understood that the present invention is not limited to the particular methodology, protocols, cell lines, animal species or genera, and reagents described, as these may vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is limited only by the appended claims. As used herein, the singular forms "a," "and," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to a "cell" includes a plurality of such cells, a reference to a "protein" includes one or more proteins and equivalents thereof known to those of skill in the art, and so forth. All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless clearly dictated otherwise.

[0107] As used herein, the term "most bothersome symptoms associated with migraine" refers to symptoms that an individual patient identifies as the most bothersome symptoms associated with his / her migraine. In the present invention, the "most bothersome symptoms associated with migraine" are identified in Table 1. The "most bothersome symptoms associated with migraine" of the present invention are described by the patient to the investigator who assists in the medical interpretation of the patient's symptoms. The investigator of the clinical trial was able to see the patient with a group of known migraine symptoms selected from the group consisting of sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), aura, pain with activity, pain, pulsation / pulsation, cognitive disruption, fatigue, mood swings, sensitivity to smell (osmophobia or olfactophobia), visual effects, pressure / tension, pain (anatomical), eye pain, neck pain, dizziness, allodynia, inactivity, sensory disturbance, sleep disturbance, and speech disturbance. A patient's "most bothersome symptoms associated with migraine", as used herein, refers to their self-identified "most bothersome symptoms associated with migraine", which may be one or more of the symptoms listed herein above, or may be classified as "other".

[0108] As used herein, the term "improvement" or "ameliorating" the most bothersome symptoms associated with migraine refers to a change in the patient's assessment of MBS as compared to baseline (i.e., the MBS prior to the first administration of an anti-CGRP antibody or fragment thereof of the invention). Improvement is characterized by a one or more category change in the patient's assessment of MBS as compared to baseline on a 7-point Likert scale as described in Example 2.

[0109] As used herein, the term "improvement" or "improving" Patient Global Impression of Migraine-related Changes refers to a change in the patient's assessment of the disease state as compared to baseline (i.e., the disease state prior to the first administration of an anti-CGRP antibody or fragment thereof of the invention). Improvement is characterized by a one or more category change in the patient's assessment of PGIC as compared to baseline on the 7-point scale described in Example 2.

[0110] As used herein, the term "chronic migraine" refers to a condition in which patients experience, on average, at least 15 headache days per month, with a subset of these headache days meeting the ICHD-3 criteria for migraine with or without aura. The term "episodic migraine" refers to a condition in which patients experience, on average, fewer than 15 headache days per month, with typically 4 to 15 days being of a migraine phenotype that meets the ICHD-3 definition of migraine with or without aura.

[0111] As used herein, the term "diagnosed with chronic migraine" refers to a patient meeting the criteria for chronic migraine, regardless of whether a formal diagnosis has been made for the patient.

[0112] As used herein, the term "administered intravenously" refers to a mode of administration in which a substance, such as an antibody, is introduced directly into the patient's circulation, most typically into the venous circulation. The substance may be introduced in a carrier fluid, such as an aqueous solution, e.g., normal saline. The substance may be administered in a single formulation or in multiple formulations, so long as administration is completed within a short period of time (e.g., within a day, preferably within 12 hours, more preferably within 6 hours, and most preferably within 1-2 hours).

[0113] As used herein, the term "baseline number of migraine days" refers to the number of migraine days a patient experiences during a particular period of time, e.g., before treatment. For example, the baseline number of migraine days can be determined over a period of one or more months, e.g., by recording whether or not a migraine occurred on each day.

[0114] As used herein, the term "headache days per month" refers to the number of days per month that a patient has a migraine, i.e., the patient has symptoms that meet the clinical definition of a migraine at any time during that day. The number of migraine days per month can be determined by recording whether or not a migraine occurred on each day.

[0115] As used herein, the term "headache days per month" refers to the number of days per month that a patient has a headache, i.e., that at any time during that day the patient has symptoms that meet the clinical definition of a headache. The number of headache days per month can be determined by recording whether or not a headache occurred on each day.

[0116] Calcitonin Gene-Related Peptide (CGRP): As used herein, CGRP includes but is not limited to the following Homo sapiens CGRP-α and Homo sapiens CGRP-β amino acid sequences available from American Peptides (Sunnyvale Calif.) and Bachem (Torrance, Calif.): CGRP-α: ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSKAF-NH2 (SEQ ID NO:561), where the terminal phenylalanine is amidated; CGRP-β: ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSKAF-NH2 (SEQ ID NO:562), where the terminal phenylalanine is amidated; any membrane-bound forms of these CGRP amino acid sequences, as well as mutants, splice variants, isoforms, orthologs, homologs and variants of this sequence.

[0117] Expression vectors: These DNA vectors contain elements that facilitate manipulation for the expression of foreign proteins in target host cells, such as yeast or mammalian cells, such as Pichia pastoris or CHO cells. Conveniently, manipulation of sequences for transformation and production of DNA is first performed in a bacterial host, such as E. coli, and the vector will usually contain sequences to facilitate such manipulation, including a bacterial origin of replication and an appropriate bacterial selection marker. A selection marker encodes a protein necessary for the survival or growth of transformed host cells grown in a selective medium. Host cells not transformed with a vector containing a selection gene will not survive in the medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, (b) complement an auxotrophic defect, or (c) supply vital nutrients not available from complex media. Exemplary vectors and methods for yeast transformation are described, for example, in Burke, D., Dawson, D., & Stearns, T. (2000). Methods in yeast genetics: a Cold Spring Harbor Laboratory course manual. Plainview, NY: Cold Spring Harbor Laboratory Press.

[0118] Expression vectors for use in yeast or mammalian cells will generally further comprise yeast or mammalian specific sequences, including selectable auxotrophic or drug markers for identifying transformed yeast strains or transformed mammalian cells. Drug markers can further be used to amplify the copy number of the vector in the host cell.

[0119] The polypeptide coding sequence of interest is operably linked to transcriptional and translational regulatory sequences that provide for expression of the polypeptide in a host cell, such as Pichia pastoris or CHO cells. These vector components may include, but are not limited to, one or more of the following: enhancer elements, promoters, and transcription termination sequences. Sequences for secretion of the polypeptide, such as a signal sequence, may also be included. A yeast or mammalian origin of replication is optional, since expression vectors often integrate into the host cell genome. In one embodiment of the present invention, the polypeptide of interest is operably linked or fused to sequences that provide for optimized secretion of the polypeptide from yeast diploid cells.

[0120] A nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a signal sequence is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence. Generally, "operably linked" means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading frame. Enhancers, however, need not be contiguous. This can be accomplished by ligation at convenient restriction sites, or via PCR / recombination methods well known to those of skill in the art (Gateway® Technology; Invitrogen, Carlsbad California). If such sites do not exist, synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.

[0121] Promoters are untranslated sequences located upstream (5') of the start codon of a structural gene (generally within about 100-1000 bp) that control the transcription and translation of a particular nucleic acid sequence to which they are operably linked. Such promoters are divided into several classes: inducible, constitutive, and repressible promoters (which increase transcription levels in response to the absence of a repressor). Inducible promoters can initiate increased levels of transcription from DNA under their control in response to some change in culture conditions, e.g., the presence or absence of a nutrient, or a change in temperature.

[0122] The promoter fragment may also serve as a site for homologous recombination and integration of the expression vector into the same site of the host genome; alternatively, a selectable marker is used as the site of homologous recombination. Examples of suitable promoters from Pichia include the AOX1 and promoters (Cregg et al. (1989) Mol. Cell. Biol. 9:1316-1323); the ICL1 promoter (Menendez et al. (2003) Yeast 20(13):1097-108); the glyceraldehyde-3-phosphate dehydrogenase promoter (GAP) (Waterham et al. (1997) Gene 186(1):37-44); and the FLD1 promoter (Shen et al. (1998) Gene 216(1):93-102). The GAP promoter is a strong constitutive promoter, and the AOX and FLD1 promoters are inducible.

[0123] Other yeast promoters include ADH1, alcohol dehydrogenase II, GAL4, PHO3, PHO5, Pyk, and chimeric promoters derived therefrom. In addition, non-yeast promoters, such as mammalian, insect, plant, reptile, amphibian, viral, and avian promoters, can be used in the present invention. Most typically, the promoter will include a mammalian promoter (which may be endogenous to the expressed gene), or a yeast or viral promoter that provides efficient transcription in yeast systems.

[0124] Examples of mammalian promoters include, among others, promoters from cytomegalovirus (CMV), promoters from chicken 3-actin (CBM), promoters from adenomatous polyposis coli (APC), leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5) promoter, CAG promoter, β-actin promoter, elongation factor-1 (EF1) promoter, early growth response 1 (EGR-1) promoter, eukaryotic initiation factor 4A (EIF4A1) promoter, simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, avian leukemia virus promoter, Epstein-Barr virus immediate early promoter, Rous sarcoma virus promoter, and human gene promoters such as, but not limited to, actin promoter, myosin promoter, hemoglobin promoter, and creatine kinase promoter. Combinations of two or more of the aforementioned promoters may be used. Additionally, inducible promoters may be used. The use of an inducible promoter provides a molecular switch that can turn on expression of an operably linked polynucleotide sequence when such expression is desired, or turn off expression when expression is not desired. Examples of inducible promoters include, but are not limited to, metallothionine promoters, glucocorticoid promoters, progesterone promoters, and tetracycline promoters.

[0125] The polypeptide of interest may also be produced as a fusion polypeptide with a heterologous polypeptide (e.g., a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide). In general, the signal sequence may be a component of the vector or may be part of the polypeptide coding sequence inserted into the vector. The heterologous signal sequence selected is preferably one that is recognized and processed through one of the standard pathways available in the host cell. The S. cerevisiae alpha-factor prepro signal has proven effective for secretion of a variety of recombinant proteins from P. pastoris. Other yeast signal sequences include the alpha-mating factor signal sequence, the invertase signal sequence, and signal sequences derived from other secreted yeast polypeptides. In addition, these signal peptide sequences can be engineered to provide enhanced secretion in diploid yeast expression systems. Secretion signals used in mammalian and yeast cells include mammalian signal sequences that may be heterologous to the protein to be secreted or may be the native sequence of the protein to be secreted. The signal sequence includes a prepeptide sequence and, in some instances, may include a propeptide sequence. Many such signal sequences are known in the art, including signal sequences found on immunoglobulin chains, e.g., the K28 preprotoxin sequence, PHA-E, FACE, human MCP-1, human serum albumin signal sequence, human Ig heavy chain, human Ig light chain, etc. See, e.g., Hashimoto et.al. Protein Eng 11(2)75(1998); and Kobayashi et.al. Therapeutic Apheresis 2(4)257(1998).

[0126] Transcription can be increased by inserting a transcription activator sequence into the vector. These activators are cis-acting elements of DNA, usually about 10-300 bp, that act on a promoter to increase its transcription. Transcriptional enhancers are relatively orientation- and position-independent and have been found 5' and 3' to transcription units, within introns, and within the coding sequence itself. Enhancers can be spliced ​​into the expression vector at positions 5' and 3' to the coding sequence, but are preferably located at a site 5' from the promoter.

[0127] Expression vectors used in eukaryotic host cells may contain sequences necessary for the termination of transcription and stabilization of the mRNA. Such sequences are commonly available 3' to the translation stop codon in the untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide fragments transcribed as polyadenylated fragments in the untranslated portion of the mRNA.

[0128] Construction of suitable vectors containing one or more of the components listed above uses standard ligation techniques or PCR / recombination methods. Isolated plasmids or DNA fragments are cleaved, tailored and religated in the desired form to generate the required plasmid or are generated via recombination methods. The ligation mixture is used to transform host cells and successful transformants are selected by antibiotic resistance (e.g., ampicillin or zeocin) where appropriate for analysis to confirm correct sequences in the constructed plasmid. Plasmids from the transformants are prepared, analyzed by restriction endonuclease digestion and / or sequenced.

[0129] As an alternative to fragment restriction and ligation, DNA sequences can be inserted into vectors using recombination methods based on att sites and recombinase enzymes. Such methods are described, for example, by Landy (1989) Ann. Rev. Biochem. 58:913-949; and are known to those skilled in the art. Such methods utilize intermolecular DNA recombination mediated by a mixture of recombination proteins encoded by lambda and E. coli. Recombination occurs between specific attachment (att) sites on interacting DNA molecules. For a description of att sites, see Weisberg and Landy (1983) Site-Specific Recombination in Phage Lambda, in Lambda II, Weisberg, ed. (Cold Spring Harbor, NY: Cold Spring Harbor Press), pp. 211-250. DNA segments flanking the recombination sites are exchanged, such that after recombination, the att sites are hybrid sequences consisting of sequences contributed by each parent vector. Recombination can occur between DNA of any topology.

[0130] Att sites can be introduced into a sequence of interest by ligating the sequence into a suitable vector; generating a PCR product containing attB sites by use of specific primers; generating a cDNA library cloned into a suitable vector containing att sites, etc.

[0131] Folding, as used herein, refers to the three-dimensional structure of polypeptides and proteins, where interactions between amino acid residues act to stabilize the structure. Proper folding is typically the configuration of a polypeptide that results in optimal biological activity, and in the case of antibodies, can be conveniently monitored by assays for activity (e.g., antigen binding).

[0132] The expression host may be further modified by the introduction of sequences encoding one or more enzymes that enhance folding and disulfide bond formation, i.e., foldases, chaperonins, etc. Such sequences may be constitutively or inducibly expressed in the yeast host cell using vectors, markers, etc. known in the art. Preferably, sequences containing sufficient transcriptional regulatory elements for the desired expression pattern are stably integrated into the yeast genome by targeted methodologies.

[0133] For example, eukaryotic PDI is not only an efficient catalyst for protein cysteine ​​oxidation and disulfide bond isomerization, but also exhibits chaperone activity. Co-expression of PDI can promote the production of active proteins with multiple disulfide bonds. Expression of BIP (immunoglobulin heavy chain binding protein); cyclophilin, etc. is also of interest. In one embodiment of the present invention, each of the monoploid parent strains expresses a different folding enzyme, for example, one strain may express BIP and the other strain may express PDI or a combination thereof.

[0134] The terms "desired protein" or "desired antibody" are used interchangeably and generally refer to a target-specific parent antibody, i.e., CGRP or a chimeric or humanized antibody as described herein, or a binding portion thereof derived therefrom. The term "antibody" is intended to include any polypeptide chain-containing molecular structure having a specific shape that matches and recognizes an epitope, where one or more non-covalent interactions stabilize the complex between the molecular structure and the epitope. The prototypic antibody molecule is an immunoglobulin, and immunoglobulins from all sources, e.g., human, rodent, rabbit, bovine, ovine, porcine, canine, other mammals, chicken, other birds, etc., particularly IgG etc., are considered to be "antibodies." The preferred source for producing antibodies useful as starting materials according to the present invention is rabbit. Numerous antibody coding sequences have been described; others can be made by methods well known in the art. Examples include chimeric antibodies, human antibodies and other non-human mammalian antibodies, humanized antibodies, single chain antibodies (e.g., scFvs), camelid antibodies, nanobodies, IgNAR (single chain antibody from shark), small modular immunopharmaceuticals (SMIPs), and antibody fragments such as Fabs, Fab', F(ab')2.See Streltsov VA,et al.,Structure of a shark IgNAR antibody variable domain and modeling of an early-developmental isotype,Protein Sci.2005 Nov;14(11):2901-9.Epub 2005 Sep 30;Greenberg AS,et al.,A new antigen receptor gene family that undergoes rearrangement and extensive somatic diversification in sharks,Nature.1995 Mar 9;374(6518):168-73;Nuttall SD,et al.,Isolation of the new antigen receptor from wobbegong sharks,and use as a scaffold for the display of protein loop libraries,Mol Immunol.2001 Aug;38(4):313-26;Hamers-Casterman C,et al.,Naturally occurring antibodies devoid of light chains,Nature.1993 Jun 3;363(6428):446-8;Gill DS,et al.,Biopharmaceutical drug discovery using novel protein scaffolds,Curr Opin Biotechnol.2006 Dec;17(6):653-8.Epub 2006 Oct 19.

[0135] For example, antibodies or antigen-binding fragments can be produced by genetic engineering. In this technique, as in other methods, antibody-producing cells are sensitized to the desired antigen or immunogen. Using messenger RNA isolated from the antibody-producing cells as a template, PCR amplification is used to generate cDNA. A library of vectors, each containing one heavy chain gene and one light chain gene that retains the original antigen specificity, is generated by inserting appropriate segments of the amplified immunoglobulin cDNA into an expression vector. A combinatorial library is constructed by combining a heavy chain gene library with a light chain gene library. This results in a library of clones that co-express heavy and light chains (similar to the Fab fragment or antigen-binding fragment of an antibody molecule). The vectors carrying these genes are co-transfected into a host cell. When antibody gene synthesis is induced in the transfected host, the heavy and light chain proteins self-assemble to produce active antibodies that can be detected by screening with the antigen or immunogen.

[0136] Antibody coding sequences of interest include those encoded by natural sequences, as well as nucleic acids that are not identical in sequence to the disclosed nucleic acids and variants thereof due to the degeneracy of the genetic code, and variants thereof. Variant polypeptides may include amino acid (aa) substitutions, additions, or deletions. Amino acid substitutions may be conservative amino acid substitutions to eliminate non-essential amino acids, e.g., to modify glycosylation sites, or to minimize misfolding by substitution or deletion of one or more cysteine ​​residues not required for function. Variants may be designed to retain or have enhanced biological activity of specific regions of the protein (e.g., functional domains, catalytic amino acid residues, etc.). Variants also include fragments of the polypeptides disclosed herein, particularly biologically active fragments and / or fragments corresponding to functional domains. Techniques of in vitro mutagenesis of cloned genes are known. Polypeptides that have been modified using routine molecular biology techniques to improve their resistance to proteolysis, or to optimize solubility properties, or to make them more suitable as therapeutic agents, are also included in the present invention.

[0137] Chimeric antibodies are antibodies that combine variable light and heavy chain regions (VCH) derived from antibody-producing cells of one species. L and V H ) with constant light and heavy chain regions from another species. Typically, chimeric antibodies utilize rodent or rabbit variable regions and human constant regions to produce an antibody with predominantly human domains. The production of such chimeric antibodies is well known in the art and can be accomplished by standard means (e.g., as described in U.S. Pat. No. 5,624,659, which is incorporated herein by reference in its entirety). It is further contemplated that the human constant region of the chimeric antibody of the present invention can be selected from IgG1, IgG2, IgG3, and IgG4 constant regions.

[0138] Humanized antibodies are engineered to contain more human-like immunoglobulin domains and incorporate only the complementarity determining regions of the animal-derived antibody. This is accomplished by carefully sequencing the hypervariable loops of the variable regions of a monoclonal antibody to fit the structure of the human antibody chains. Although superficially complex, the process is straightforward in practice. See, for example, U.S. Patent No. 6,187,287, which is incorporated herein by reference in its entirety.

[0139] In addition to whole immunoglobulins (or their recombinant counterparts), immunoglobulin fragments (e.g., Fab', F(ab')2, or other fragments) that contain epitope binding sites can be synthesized. "Fragments", or minimal immunoglobulins, can be engineered using recombinant immunoglobulin technology. For example, "Fv" immunoglobulins for use in the present invention can be produced by synthesizing fused variable light and variable heavy chain regions. Combinations of antibodies, such as diabodies that contain two different Fv specificities, are also of interest. In another embodiment of the present invention, SMIPs (small molecule immunopharmaceuticals), camelid antibodies, nanobodies, and IgNARs are encompassed by immunoglobulin fragments.

[0140] Immunoglobulins and fragments thereof may be post-translationally modified to add effector moieties such as, for example, chemical linkers, detectable moieties such as fluorochromes, enzymes, toxins, substrates, bioluminescent materials, radioactive materials, chemiluminescent moieties, or specific binding moieties such as streptavidin, avidin, or biotin, or may be used in the methods and compositions of the invention. Examples of additional effector molecules are provided below.

[0141] A polynucleotide sequence "corresponds to" a polypeptide sequence (i.e., the polynucleotide sequence "encodes" a polypeptide sequence) if translation of the polynucleotide sequence according to the genetic code results in the polypeptide sequence, and one polynucleotide sequence "corresponds to" another polynucleotide sequence if the two sequences encode the same polypeptide sequence.

[0142] A "heterologous" region or domain of a DNA construct is an identifiable segment of DNA within a larger DNA molecule that is not found in association with the larger naturally occurring molecule. Thus, if a heterologous region encodes a mammalian gene, the gene will usually be flanked by DNA that does not flank the mammalian genomic DNA in the genome of the source organism. Another example of a heterologous region is a construct in which the coding sequence itself is not found in nature (e.g., a cDNA in which the genomic coding sequence contains introns, or a synthetic sequence with codons different from the native gene). Allelic variations or naturally occurring mutational events do not give rise to a heterologous region of DNA as defined herein.

[0143] A "coding sequence" is an in-frame sequence of codons that corresponds to a protein or peptide sequence (in terms of the genetic code). Two coding sequences correspond to each other if the sequences or their complementary sequences code for the same amino acid sequence. A coding sequence in association with appropriate regulatory sequences can be transcribed and translated into a polypeptide. A polyadenylation signal and transcription termination sequence will usually be located 3' to the coding sequence. A "promoter sequence" is a DNA regulatory region that is capable of binding RNA polymerase in a cell and initiating transcription of a downstream (3' direction) coding sequence. Promoter sequences typically contain additional sites for the binding of regulatory molecules (e.g., transcription factors) that affect transcription of the coding sequence. A coding sequence is "under the control" of a promoter sequence or "operably linked" to a promoter when RNA polymerase binds to the promoter sequence in a cell and transcribes the coding sequence into mRNA, which is then translated into the protein encoded by the coding sequence.

[0144] Vectors are used to introduce foreign substances, such as DNA, RNA or proteins, into organisms or host cells. Typical vectors include recombinant viruses (for polynucleotides) and liposomes (for polypeptides). A "DNA vector" is a replicon, such as a plasmid, phage, or cosmid, to which another polynucleotide segment can be attached so as to bring about the replication of the attached segment. An "expression vector" is a DNA vector that contains regulatory sequences that direct polypeptide synthesis by an appropriate host cell. This usually means that a promoter binds RNA polymerase and initiates transcription of mRNA, and that a ribosome binding site and initiation signal direct translation of the mRNA into a polypeptide. Incorporation of a polynucleotide sequence into an expression vector at the appropriate site and in the correct reading frame, followed by transformation of an appropriate host cell with the vector, allows for the production of the polypeptide encoded by the polynucleotide sequence.

[0145] "Amplification" of a polynucleotide sequence is the in vitro production of multiple copies of a particular nucleic acid sequence. The amplified sequence is usually in the form of DNA. Various techniques for carrying out such amplification are described in a review article by Van Brunt (1990, Bio / Technol., 8(4):291-294). The polymerase chain reaction, or PCR, is the prototype of nucleic acid amplification, and the use of PCR herein should be considered as illustrative of other suitable amplification techniques.

[0146] The general structure of antibodies in vertebrates is now well understood (Edelman, GM, Ann. NY Acad. Sci., 190:5 (1971)). Antibodies consist of two identical light polypeptide chains ("light chains") of molecular weight approximately 23,000 daltons, and two identical heavy chains ("heavy chains") of molecular weight 53,000-70,000. The four chains are linked by disulfide bonds in a "Y" configuration, with the light chains enveloping the heavy chains beginning at the mouth of the "Y" configuration. The "branched" portion of the "Y" configuration is the F ab The stem of the "Y" configuration is called the FC The orientation of the amino acid sequences runs from the N-terminus at the top of the "Y" configuration to the C-terminus at the bottom of each chain. The N-terminus contains the variable region with specificity for the antigen that elicited it, which is about 100 amino acids in length and has slight variation between light and heavy chains and from antibody to antibody.

[0147] The variable region is linked in each chain to a constant region that extends the remaining length of the chain; that of a particular class of antibody does not vary depending on the specificity of the antibody (i.e., the antigen that elicited it). There are five known major classes of constant regions that determine the class of immunoglobulin molecule (IgG, IgM, IgA, IgD, and IgE, corresponding to the γ, μ, α, δ, and ε (gamma, mu, alpha, delta, or epsilon) heavy chain constant regions). The constant region or class determines the subsequent effector functions of the antibody, including activation of complement (Kabat, EA, Structural Concepts in Immunology and Immunochemistry, 2nd Ed., p. 413-436, Holt, Rinehart, Winston (1976)), and other cellular responses (Andrews, DW, et al., Clinical Immunobiology, pp 1-18, WB Sanders (1980); Kohl, S., et al., Immunology, 48:187 (1983)); while the variable region determines the antigen with which it reacts. Light chains are classified as either κ (kappa) or λ (lambda). Each heavy chain class can be prepared with either kappa or lambda light chains. The light and heavy chains are covalently linked to each other, and when an immunoglobulin is produced either by a hybridoma or a B cell, the "tail" portions of the two heavy chains are linked to each other by a covalent disulfide bond.

[0148] The term "variable region" or "VR" refers to the domains within each pair of light and heavy chains of an antibody that are directly involved in binding the antibody to an antigen. Each heavy chain contains a variable domain (VR) at one end. H ) followed by a number of constant domains. Each light chain has a variable domain (V Lat its other end a constant domain; the constant domain of the light chain is aligned with the first constant domain of the heavy chain and the light chain variable domain is aligned with the variable domain of the heavy chain.

[0149] The terms "complementarity determining region", "hypervariable region" or "CDR" refer to one or more of the hypervariable or complementarity determining regions (CDRs) found in the variable region of the light or heavy chain of an antibody (see Kabat, EA et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1987)). These expressions include the hypervariable regions as defined by Kabat et al. ("Sequences of Proteins of Immunological Interest," Kabat E., et al., US Dept. of Health and Human Services, 1983), or the hypervariable loops in the three-dimensional structure of an antibody (Chothia and Lesk, J Mol. Biol. 196 901-917 (1987)). The CDRs of each chain are held in close proximity by framework regions and, together with the CDRs from the other chain, contribute to the formation of the antigen binding site. Within the CDRs are select amino acids that have been described as selectivity determining regions (SDRs) that represent important contact residues used by the CDRs in antibody-antigen interactions (Kashmiri, S., Methods, 36:25-34 (2005)). In the present invention, when a particular antibody amino acid or nucleic acid residue is referred to by a number, this generally refers to its position within a particular amino acid or nucleic acid sequence (i.e., a particular sequence identifier) ​​and / or its position according to the numbering system of Kabat et al.

[0150] The term "framework region" or "FR" refers to one or more of the framework regions in the variable region of an antibody light or heavy chain (see Kabat, EA et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1987)). These terms include the amino acid sequence regions intervening between the CDRs in the variable regions of the light and heavy chains of an antibody.

[0151] "Cmax" refers to the maximum (or peak) concentration that an antibody or other compound achieves in a test area (e.g., serum or another compartment such as cerebrospinal fluid) after a drug is administered. For example, serum Cmax can be measured from serum, which is prepared, for example, by collecting a blood sample, allowing it to clot, and separating the solid components by centrifugation or other means to produce serum (blood without blood cells or clotting factors), and then detecting the concentration of an analyte in the serum by ELISA or other means known in the art.

[0152] "AUC" refers to the area under the concentration-time curve, expressed in units of mg / mL x hr (or equivalently, mg x hr / ml), unless otherwise specified. 0-t " refers to the area under the concentration-time curve from time = 0 to the last quantifiable concentration. "AUC 0-inf " refers to the area under the concentration-time curve extrapolated from time = 0 to infinity.

[0153] "I max " refers to the maximal pharmacodynamic response elicited by an anti-CGRP antibody dose, preferably 350 mg or more, more typically at least 750 or 1000 mg, compared to the response elicited by a lower anti-CGRP antibody dose; for example, such a response may be detected by inhibition of vasodilation following topical application of capsaicin.

[0154] Anti-CGRP antibodies and their binding fragments having binding specificity for CGRP The present invention specifically includes the use of a particular anti-CGRP antibody or antibody fragment, Ab6, which comprises or consists of the CDR, VL, VH, CL, CH polypeptide sequences identified in Figures 1 to 12. Polypeptides included in the anti-CGRP antibody, Ab6, are further described below.

[0155] Antibody Ab6 In a preferred exemplary embodiment, the invention includes a humanized antibody that has binding specificity for CGRP and has a variable light chain sequence comprising the sequence set forth below: QVLTQSPSSLSASVGDRVTINCQASQSVYHNTYLAWYQQKPGKVPKQLIYDASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCTNGDCFVFGGGTKVEIKR (SEQ ID NO: 222)

[0156] The present invention also includes a humanized antibody that has binding specificity for CGRP and has a light chain sequence comprising the sequence set forth below: [ka]

[0157] The present invention also includes a humanized antibody that has binding specificity for CGRP and has a variable heavy chain sequence comprising the sequence set forth below: EVQLVESGGGLVQPGGSLRLSCAVSGIDLSGYYMNWVRQAPGKGLEWVGVIGINGATYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSS (SEQ ID NO: 202)

[0158] The present invention also includes a humanized antibody that has binding specificity for CGRP and has a heavy chain sequence comprising the sequence set forth below: [ka]

[0159] Alternatively, the heavy chain of Ab6 may lack the C-terminal lysine of SEQ ID NO:201, i.e., the heavy chain sequence comprises the sequence set forth below: [ka]

[0160] The invention further contemplates an antibody comprising one or more of the polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, which correspond to the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of SEQ ID NO:222 or the light chain sequence of SEQ ID NO:221, and / or one or more of the polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, which correspond to the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO:202 or the heavy chain sequence of SEQ ID NO:201 or SEQ ID NO:566, or a combination of these polypeptide sequences. In another embodiment of the invention, the antibody or fragment thereof of the invention comprises or consists of one or more of the CDRs, variable heavy and variable light chain sequences, and combinations of the heavy and light chain sequences described above (including all of them).

[0161] The present invention also contemplates fragments of antibodies that have binding specificity for CGRP. In one embodiment of the invention, an antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 222 or SEQ ID NO: 221. In another embodiment of the invention, an antibody fragment of the invention comprises or consists of the polypeptide sequence of SEQ ID NO: 202 or SEQ ID NO: 201 or SEQ ID NO: 566.

[0162] In further embodiments of the invention, antibody fragments with binding specificity for CGRP comprise, or consist of, one or more of the polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, which correspond to the complementarity determining regions (CDRs, or hypervariable regions) of the variable light chain sequence of SEQ ID NO:222 or the light chain sequence of SEQ ID NO:221.

[0163] In further embodiments of the invention, antibody fragments with binding specificity for CGRP comprise, or consist of, one or more of the polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, which correspond to the complementarity determining regions (CDRs, or hypervariable regions) of the variable heavy chain sequence of SEQ ID NO:202, or the heavy chain sequence of SEQ ID NO:201 or SEQ ID NO:566.

[0164] The present invention also contemplates antibody fragments comprising one or more of the antibody fragments described herein. In one embodiment of the present invention, the fragment of an antibody that has binding specificity for CGRP comprises, or consists of, one, two, three or more, including all of the following antibody fragments: the variable light chain region of SEQ ID NO:222; the variable heavy chain region of SEQ ID NO:202; the complementarity determining regions of the variable light chain region of SEQ ID NO:222 (SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228); and the complementarity determining regions of the variable heavy chain region of SEQ ID NO:202 (SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208).

[0165] In a particularly preferred embodiment of the invention, the humanized anti-CGRP antibody is Ab6, comprising or consisting of SEQ ID NO:221 and SEQ ID NO:201 or SEQ ID NO:566 and having at least one of the biological activities described herein.

[0166] In a further particularly preferred embodiment of the invention, the antibody fragment comprises or consists of a Fab (fragment antigen binding) fragment that has binding specificity for CGRP. In the context of the antibody Ab6, the Fab fragment comprises the variable light chain region of SEQ ID NO: 222 and the variable heavy chain region of SEQ ID NO: 202. This embodiment of the invention further contemplates additions, deletions and variants of SEQ ID NO: 222 and / or SEQ ID NO: 202 in said Fab whilst retaining binding specificity for CGRP.

[0167] In another particularly preferred embodiment of the invention, the anti-CGRP antibody may comprise an antibody expression product isolated from a recombinant cell expressing a nucleic acid sequence encoding the variable light chain polypeptide of SEQ ID NO:222 and the variable heavy chain polypeptide of SEQ ID NO:202, which are optionally linked to human light and heavy chain constant region polypeptides, respectively, such as human IgG1, IgG2, IgG3 or IgG4 constant regions, which may optionally be modified to alter glycosylation or proteolysis, and wherein the recombinant cell optionally comprises a yeast or mammalian cell, such as a Pichia pastoris or CHO cell.

[0168] In another particularly preferred embodiment of the invention, the anti-CGRP antibody can comprise an antibody expression product isolated from a recombinant cell expressing a nucleic acid sequence encoding a light chain polypeptide of SEQ ID NO:221 and a heavy chain polypeptide of SEQ ID NO:201 or SEQ ID NO:566, the recombinant cell optionally comprising a yeast or mammalian cell, e.g., a Pichia pastoris or a CHO cell, the constant region of which can optionally be modified to alter glycosylation or proteolysis or other effector function.

[0169] In another particularly preferred embodiment of the invention, any of the aforementioned anti-CGRP antibodies or antibody fragments may be included in a formulation disclosed herein, e.g., comprising histidine (L-histidine), sorbitol, polysorbate 80, e.g., about 100 mg of anti-CGRP antibody, about 3.1 mg of L-histidine, about 40.5 mg of sorbitol, and about 0.15 mg of polysorbate 80 per mL volume, having a pH of about 5.8.

[0170] In one embodiment of the invention described herein (below), Fab fragments may be produced by enzymatic digestion of Ab6 (e.g., papain). In another embodiment of the invention, anti-CGRP antibodies, such as Ab6 or Fab fragments thereof, may be produced via expression in mammalian cells, such as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systems, such as yeast cells (e.g., diploid yeast, e.g., diploid Pichia) and other yeast strains. Suitable Pichia species include, but are not limited to, Pichia pastoris.

[0171] In another embodiment, the antibody fragment may exist in one or more of the following non-limiting forms: Fab, Fab', F(ab')2, Fv, and single chain Fv antibody formats. In a preferred embodiment, the anti-CGRP antibodies described herein further comprise a kappa constant light chain sequence comprising the sequence set forth below: VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 563)

[0172] In another preferred embodiment, the anti-CGRP antibody described herein further comprises a gamma-1 constant heavy chain polypeptide sequence comprising the sequence set forth below, or the same sequence lacking the carboxy-terminal lysine residue (SEQ ID NO:564 and SEQ ID NO:565, respectively): [ka]

[0173] For clarity, any antibody disclosed herein is intended to include any variant of the constant region variant sequences disclosed, for example, Ab6 may include the constant region of SEQ ID NO: 564 including a C-terminal lysine, or may include the constant region of SEQ ID NO: 565 lacking the C-terminal lysine. Thus, all disclosures herein of the heavy chain of SEQ ID NO: 201 also include variants lacking its C-terminal lysine residue, i.e., having the heavy chain variable region sequence of Ab6 (SEQ ID NO: 202) and the constant region sequence of SEQ ID NO: 565. For example, a sequence encoding an antibody that includes a C-terminal lysine in its heavy chain, when expressed in a cell line, e.g., CHO cells, may produce an antibody lacking said C-terminal lysine due to proteolysis, or a mixture of heavy chains that either include or lack said C-terminal lysine.

[0174] In one embodiment of the present invention, an antibody or V H Or V L Polypeptides are generated or selected from one or more rabbit B cell populations prior to initiation of the humanization process referred to herein.

[0175] In another embodiment of the invention, the anti-CGRP antibodies and fragments thereof have no binding specificity for CGRP-R. In a further embodiment of the invention, the anti-CGRP antibodies and fragments thereof inhibit the association of CGRP with CGRP-R. In another embodiment of the invention, the anti-CGRP antibodies and fragments thereof inhibit the association of CGRP with CGRP-R and / or additional proteins and / or multimers thereof and / or antagonize the biological effect thereof.

[0176] As described herein, antibodies and fragments thereof may be post-translationally modified to attach effector moieties, such as chemical linkers, detectable moieties, e.g., fluorochromes, enzymes, substrates, bioluminescent materials, radioactive materials, and chemiluminescent moieties, or functional moieties, e.g., streptavidin, avidin, biotin, cytotoxins, cytotoxic materials, and radioactive materials.

[0177] Antibodies or fragments thereof may also be chemically modified to provide additional advantages, such as increasing the solubility, stability and circulation time (in vivo half-life) of the polypeptide, or decreasing immunogenicity (see U.S. Pat. No. 4,179,337). Chemical moieties for derivatization may be selected from water-soluble polymers, such as polyethylene glycol, ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, and the like. Antibodies and fragments thereof may be modified at random positions within the molecule or at predetermined positions within the molecule and may contain one, two, three or more attached chemical moieties.

[0178] The polymer may be of any molecular weight, branched or unbranched. For polyethylene glycol, the preferred molecular weight is about 1 kD to about 100 kDa for ease of handling and manufacturing (the term "about" indicates that in a preparation of polyethylene glycol, some molecules will be heavier and some will be lighter than the stated molecular weight). Other sizes can be used depending on the desired therapeutic profile (e.g., the desired duration of sustained release, the effect on biological activity (if any), ease of handling, the degree or lack of antigenicity, and other known effects of polyethylene glycol on the therapeutic protein or analogue). For example, polyethylene glycol may be about 200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, It may have an average molecular weight of 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000, 25,000, 30,000, 35,000, 40,000, 50,000, 55,000, 60,000, 65,000, 70,000, 75,000, 80,000, 85,000, 90,000, 95,000, or 100,000 kDa. Branched polyethylene glycols are described, for example, in U.S. Pat. No. 5,643,575; Morpurgo et al., Appl. Biochem. Biotechnol. 56:59-72 (1996); Vorobjev et al., Nucleosides Nucleotides 18:2745-2750 (1999); and Caliceti et al., Bioconjug. Chem. 10:638-646 (1999), the disclosures of each of which are incorporated herein by reference.

[0179] There are numerous methods of attachment available to those skilled in the art. See, for example, European Patent No. 0 401 384 (linking PEG to G-CSF), which is incorporated herein by reference. See also Malik et al., Exp. Hematol. 20:1028-1035 (1992) (reporting pegylation of GM-CSF using tresyl chloride). For example, polyethylene glycol can be covalently attached to amino acid residues via reactive groups such as free amino or carboxyl groups. Reactive groups are those to which an activated polyethylene glycol molecule can be attached. Amino acid residues with free amino groups can include lysine residues and N-terminal amino acid residues; amino acid residues with free carboxyl groups can include aspartic acid residues, glutamic acid residues and C-terminal amino acid residues. Sulfhydryl groups can also be used as reactive groups to attach polyethylene glycol molecules. Preferred for therapeutic purposes is attachment at amino groups, e.g., at the N-terminus or lysine groups.

[0180] As alluded to above, polyethylene glycol can be attached to proteins via linkage to any of a number of amino acid residues. For example, polyethylene glycol can be linked to polypeptides via covalent bonds to lysine, histidine, aspartic acid, glutamic acid, or cysteine ​​residues. One or more reaction chemistries may be used to attach polyethylene glycol to specific amino acid residues (e.g., lysine, histidine, aspartic acid, glutamic acid, or cysteine) or to two or more types of amino acid residues (e.g., lysine, histidine, aspartic acid, glutamic acid, cysteine, and combinations thereof).

[0181] Alternatively, the antibodies or fragments thereof may have increased in vivo half-life via fusion with albumin (including but not limited to recombinant human serum albumin or fragments or variants thereof (see, e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999; European Patent No. 0 413 622; and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, which are incorporated herein by reference in their entireties)) or other circulating blood proteins, such as transferrin or ferritin. In a preferred embodiment, the polypeptides and / or antibodies of the invention (including fragments or variants thereof) are fused to the mature form of human serum albumin (i.e., amino acids 1-585 of human serum albumin as shown in Figures 1 and 2 of European Patent No. 0 322 094, which are incorporated herein by reference in their entireties). Polynucleotides encoding the fusion proteins of the invention are also encompassed by the invention.

[0182] With respect to detectable moieties, further exemplary enzymes include, but are not limited to, horseradish peroxidase, acetylcholinesterase, alkaline phosphatase, β-galactosidase, and luciferase. Further exemplary fluorescent materials include, but are not limited to, rhodamine, fluorescein, fluorescein isothiocyanate, umbelliferone, dichlorotriazinylamine, phycoerythrin, and dansyl chloride. Further exemplary chemiluminescent moieties include, but are not limited to, luminol. Further exemplary bioluminescent materials include, but are not limited to, luciferin and aequorin. Further exemplary radioactive materials include iodine-125 ( 125 I), Carbon-14 ( 14 C), sulfur 35( 35 S), tritium ( 3 H) and phosphorus 32 ( 32 P), but are not limited to these.

[0183] With respect to functional moieties, exemplary cytotoxic agents include methotrexate, aminopterin, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine; alkylating agents such as mechlorethamine, thioepa, chlorambucil, melphalan, carmustine (BSNU), mitomycin C, lomustine (CCNU), 1-methylnitrosourea, cyclothosphamide, mechlorethamine, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiamine Platinum(II) (DDP) cisplatin and carboplatin (paraplatin); anthracyclines include daunorubicin (formerly daunomycin), doxorubicin (adriamycin), detorubicin, carminomycin, idarubicin, epirubicin, mitoxantrone and bisantrene; antibiotics include dactinomycin (actinomycin D), bleomycin, calicheamicin, mithramycin, and anthramycin (AMC); and mitotic inhibitors, such as, but not limited to, the vinca alkaloids, vincristine and vinblastine. Other cytotoxic agents include paclitaxel (taxol), ricin, Pseudomonas exotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide, emetine, etoposide, tenoposide, colchicine, dihydroxyanthracin dione, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P'-(DDD)), interferons, and mixtures of these cytotoxic agents.

[0184] Additional cytotoxic agents include, but are not limited to, chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel, gemcitabine, calicheamicin, doxorubicin, 5-fluorouracil, mitomycin C, actinomycin D, cyclophosphamide, vincristine, and bleomycin. Toxic enzymes from plants and bacteria, such as ricin, diphtheria toxin, and Pseudomonas toxin, can be conjugated to humanized or chimeric antibodies, or binding fragments thereof, to generate cell-type specific killing reagents (Youle, et al., Proc. Nat'l Acad. Sci. USA 77:5483 (1980); Gilliland, et al., Proc. Nat'l Acad. Sci. USA 77:4539 (1980); Krolick, et al., Proc. Nat'l Acad. Sci. USA 77:5419 (1980)).

[0185] Other cytotoxic agents include cytotoxic ribonucleases, such as those described by Goldenberg in U.S. Patent No. 6,653,104. Embodiments of the present invention also relate to radioimmunoconjugates in which an alpha- or beta-particle-emitting radionuclide is stably bound to an antibody, or a binding fragment thereof, with or without the use of a complexing agent. Such radionuclides include β-emitters, such as phosphorus-32 ( 32 P), Scandium-47( 47 Sc), Copper-67( 67 Cu), Gallium-67( 67 Ga), Yttrium-88( 88 Y), Yttrium-90 ( 90 Y), iodine-125( 125 I), iodine-131( 131 I), Samarium-153( 153 Sm), Lutetium-177( 177 Lu), Rhenium-186( 186 Re) or rhenium-188( 188 Re), and α-emitters such as astatine-211 ( 211 At), lead-212( 212 Pb), Bismuth-212(212 Bi) or -213( 213 Bi) or Actinium-225 ( 225 Ac) is included.

[0186] Methods for conjugating antibodies or binding fragments thereof to detectable moieties, etc., are known in the art, such as those described by Hunter et al., Nature 144:945 (1962); David et al., Biochemistry 13:1014 (1974); Pain et al., J. Immunol. Meth. 40:219 (1981); and Nygren, J., Histochem. and Cytochem. 30:407 (1982).

[0187] The embodiments described herein further include variants and equivalents that are substantially homologous to the antibodies, antibody fragments, diabodies, SMIPs, camelid antibodies, nanobodies, IgNARs, polypeptides, variable regions and CDRs described herein. These may include, for example, conservative substitution mutations (i.e., the replacement of one or more amino acids with similar amino acids). For example, conservative substitutions refer to the replacement of amino acids within the same general class, e.g., one acidic amino acid with another acidic amino acid, one basic amino acid with another basic amino acid, or one neutral amino acid with another neutral amino acid. What is intended by conservative amino acid substitutions is well known in the art.

[0188] In another embodiment, the present invention contemplates a polypeptide sequence having at least 90% or more sequence identity to any one or more of the polypeptide sequences of the antibody fragments, variable regions and CDRs described herein. More preferably, the present invention contemplates a polypeptide sequence having at least 95% or more sequence identity, even more preferably at least 98% or more sequence identity, and even more preferably at least 99% or more sequence identity to any one or more of the polypeptide sequences of the antibody fragments, variable regions and CDRs described herein. Methods for determining homology between nucleic acid and amino acid sequences are well known to those of skill in the art.

[0189] In another embodiment, the present invention further contemplates the above polypeptide homologs of the antibody fragments, variable regions and CDRs described herein, which further have anti-CGRP activity, non-limiting examples of which are described herein.

[0190] The present invention further contemplates therapeutic methods in which one or more of the anti-human CGRP antibodies discussed above are non-glycosylated, or if glycosylated, are only mannosylated; contain an Fc region modified to alter effector function, half-life, proteolysis, and / or glycosylation; are human, humanized, single chain, or chimeric; and are humanized antibodies derived from a rabbit (parent) anti-human CGRP antibody. Exemplary mutations that impair glycosylation include mutations of the Asn residue at position 297 of an IgG heavy chain constant region, such as IgG1, to another amino acid, such as Ala, as described in U.S. Patent No. 5,624,821, the entirety of which is incorporated herein by reference.

[0191] The invention further contemplates one or more anti-human CGRP antibodies, wherein the framework regions (FRs) in the variable light and variable heavy regions of the antibodies, respectively, are either unaltered or are human FRs modified by replacement of one or more human FR residues in the variable light or heavy chain regions with corresponding FR residues of a parent rabbit antibody, said human FRs being derived from human variable heavy and light chain antibody sequences selected from a library of human germline antibody sequences based on their high level of homology to the corresponding rabbit variable heavy or light chain regions compared to other human germline antibody sequences contained in the library.

[0192] The present invention also contemplates that a method of treatment may include administration of two or more anti-CGRP antibodies or fragments thereof disclosed herein. When two or more antibodies are administered to a patient, the antibodies may be administered simultaneously or concurrently, or may be staggered in their administration. The anti-CGRP activity of the anti-CGRP antibodies and fragments thereof of the present invention with binding specificity for CGRP may also be described by their binding strength or their affinity for CGRP. In one embodiment of the present invention, the anti-CGRP antibodies and fragments thereof of the present invention with binding specificity for CGRP are administered in a concentration of 5x10 -7 M, 10 -7 M, 5x10 -8 M, 10 -8 M, 5x10 -9 M, 10 -9 M, 5x10 -10 M, 10 -10 M, 5x10 -11 M, 10 -11 M, 5x10 -12 M, 10 -12 M, 5x10 -13 M or 10 -13 The dissociation constant (K D ) binds to CGRP. Preferably, the anti-CGRP antibodies and fragments thereof bind to CGRP at 10 -11 M, 5x10 -12 M or 10 -12 It binds to CGRP with a dissociation constant of about M. In certain embodiments of the invention, the anti-CGRP antibody is an Ab6 with a dissociation constant of 10 pM or less, e.g., 2-8 pM, e.g., 3-6 pM, e.g., about 5 pM or less, as measured using surface plasmon resonance (Misura, K et al, July 2019, Poster P220LB, AHS 61st Annual Scientific Meeting). In another embodiment of the invention, the anti-CGRP antibodies and fragments thereof of the invention having binding specificity for CGRP bind to linear or conformational CGRP epitopes.

[0193] In another embodiment of the invention, the anti-CGRP activity of the anti-CGRP antibodies and fragments thereof of the invention that have binding specificity for CGRP is greater than or equal to 10 -4 S -1 , 5x10 -5 S-1 , 10 -5 S -1 , 5x10 -6 S -1 , 10 -6 S -1 , 5x10 -7 S -1 , or 10 -7 S -1 In certain embodiments of the invention, the anti-CGRP antibody binds to CGRP with an off-rate of 5×10 as measured using surface plasmon resonance. -6 S -1 For example, 4×10 -6 S -1 For example, 3×10 -6 S -1 For example, 2×10 -6 S - below 1 , e.g. 1×10 -6 S -1 Ab6 with the following off rates:

[0194] Polynucleotides encoding anti-CGRP antibody polypeptides As noted above, the present invention specifically includes the use of a particular anti-CGRP antibody or antibody fragment, designated herein as Ab6, which comprises or consists of the CDR, VL, VH, CL, and CH polypeptides having the sequences specified in Figures 1-12. Nucleic acid sequences encoding the VL, VH, CL, and CH polypeptides contained in Ab6 described above are also included in Figures 1-12. Nucleic acid sequences encoding the CDR, VL, VH, CL, and CH polypeptides of a particularly preferred anti-CGRP antibody, Ab6, are further described below.

[0195] Polynucleotide encoding antibody Ab6 The present invention further relates to polynucleotides encoding antibody polypeptides that have binding specificity for CGRP. In one embodiment of the invention, a polynucleotide of the invention comprises or consists of the following polynucleotide sequence, which encodes the variable light chain polypeptide sequence of SEQ ID NO: 222: [ka]

[0196] In one embodiment of the invention, a polynucleotide of the invention comprises or consists of the following polynucleotide sequence encoding the light chain polypeptide sequence of SEQ ID NO: 221: [ka]

[0197] In another embodiment of the invention, the polynucleotide of the invention comprises or consists of the following polynucleotide sequence encoding the variable heavy chain polypeptide sequence of SEQ ID NO: 202: [ka]

[0198] In one embodiment of the invention, a polynucleotide of the invention comprises or consists of the following polynucleotide sequence encoding the heavy chain polypeptide sequence of SEQ ID NO:201: [ka]

[0199] In one embodiment of the invention, a polynucleotide of the invention comprises or consists of the following polynucleotide sequence encoding the heavy chain polypeptide sequence of SEQ ID NO:566: [ka]

[0200] In further embodiments of the invention, the polynucleotide encoding the antibody fragment with binding specificity for CGRP comprises, or consists of, one or more of the polynucleotide sequences of SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the light chain variable sequence of SEQ ID NO:222 or the light chain sequence of SEQ ID NO:221.

[0201] In further embodiments of the invention, the polynucleotide encoding the antibody fragment with binding specificity for CGRP comprises, or consists of, one or more of the polynucleotide sequences of SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, which correspond to polynucleotides encoding the complementarity determining regions (CDRs, or hypervariable regions) of the heavy chain variable sequence of SEQ ID NO:202 or the heavy chain sequence of SEQ ID NO:201 or SEQ ID NO:566.

[0202] The present invention also contemplates polynucleotide sequences comprising one or more of the polynucleotide sequences encoding the antibody fragments described herein. In one embodiment of the present invention, the polynucleotide encoding the antibody fragment with binding specificity for CGRP comprises or consists of one, two, three or more of the following polynucleotides encoding the antibody fragment: polynucleotide SEQ ID NO:232 encoding the light chain variable sequence of SEQ ID NO:222; polynucleotide SEQ ID NO:231 encoding the light chain sequence of SEQ ID NO:221; polynucleotide SEQ ID NO:212 encoding the heavy chain variable region of SEQ ID NO:202; polynucleotide SEQ ID NO:211 encoding the heavy chain sequence of SEQ ID NO:201; polynucleotide SEQ ID NO:567 encoding the heavy chain sequence of SEQ ID NO:566; polynucleotides encoding the light chain variable sequence of SEQ ID NO:222 or the complementarity determining regions of the light chain sequence of SEQ ID NO:221 (SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238); and polynucleotides encoding the heavy chain variable sequence of SEQ ID NO:202 or the complementarity determining regions of the heavy chain sequence of SEQ ID NO:201 or SEQ ID NO:566 (SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218).

[0203] In a preferred embodiment of the invention, the polynucleotide of the invention comprises or consists of a polynucleotide encoding a Fab (fragment antigen binding) fragment with binding specificity for CGRP. With reference to the antibody Ab6, a polynucleotide encoding the full length Ab6 antibody comprises or consists of polynucleotide SEQ ID NO:231 encoding the light chain sequence of SEQ ID NO:221 and polynucleotide SEQ ID NO:211 encoding the heavy chain sequence of SEQ ID NO:201 or polynucleotide SEQ ID NO:567 encoding the heavy chain sequence of SEQ ID NO:566.

[0204] Another embodiment of the invention contemplates these polynucleotides incorporated into an expression vector for expression in mammalian cells, such as CHO, NSO, HEK-293, or fungal, insect, or microbial systems, such as yeast cells, such as the yeast Pichia. Suitable Pichia species include, but are not limited to, Pichia pastoris. In one embodiment of the invention described herein (below), Fab fragments can be produced by enzymatic digestion (e.g., papain) of Ab6 after expression of the full-length polynucleotide in a suitable host. In another embodiment of the invention, anti-CGRP antibodies, such as Ab6 or Fab fragments thereof, can be produced via expression of Ab6 polynucleotides in mammalian cells, such as CHO, NSO, or HEK 293 cells, fungal, insect, or microbial systems, such as yeast cells (e.g., diploid yeast, e.g., diploid Pichia) and other yeast strains. Suitable Pichia species include, but are not limited to, Pichia pastoris.

[0205] Host cells and vectors containing the polynucleotides are also contemplated.

[0206] The present invention further contemplates vectors comprising the polynucleotides encoding the variable heavy and light chain polypeptide sequences and the individual complementarity determining regions (CDRs, or hypervariable regions) described herein, as well as host cells comprising said vector sequences. In one embodiment of the invention, the host cell is a yeast cell. In another embodiment of the invention, the yeast host cell belongs to the genus Pichia.

[0207] Methods for Producing Antibodies and Fragments Thereof In another embodiment, the present invention contemplates a method for producing anti-CGRP antibodies and fragments thereof. Methods for producing antibodies and fragments thereof secreted from polyploid, preferably diploid or tetraploid strains of mating competent yeast are taught, for example, in U.S. Patent Application Publication No. 2009 / 0022659 to Olson et al. and U.S. Patent No. 7,935,340 to Garcia-Martinez et al., the disclosures of each of which are incorporated herein by reference in their entirety. Methods for producing antibodies and fragments thereof in mammalian cells, e.g., CHO cells, are further well known in the art.

[0208] Other methods for producing antibodies are also well known to those skilled in the art. For example, methods for producing chimeric antibodies are now well known in the art (see, e.g., U.S. Patent No. 4,816,567 to Cabilly et al.; Morrison et al., PNASUSA, 81:8651-55 (1984); Neuberger, MS et al., Nature, 314:268-270 (1985); Boulianne, GL et al., Nature, 312:643-46 (1984) (the disclosures of each of which are incorporated herein by reference in their entirety).

[0209] Similarly, other methods for producing humanized antibodies are now well known in the art (e.g., U.S. Pat. Nos. 5,530,101, 5,585,089, 5,693,762, and 6,180,370 to Queen et al; U.S. Pat. Nos. 5,225,539 and 6,548,640 to Winter; U.S. Pat. Nos. 6,054,297, 6,407,213, and 6,639,055 to Carter et al; U.S. Pat. No. 6,632,927 to Adair; Jones, PT et al, Nature, 321:522-525 (1986); Reichmann, L., et al, Nature, 332:323-327 (1988); Verhoeyen, M, et al, Nature, 332:323-327 (1988); al, Science, 239:1534-36 (1988), the disclosures of each of which are incorporated herein by reference in their entireties.

[0210] The invention further includes the use of any of the pharmaceutical formulations disclosed herein in the manufacture of a medicament for the treatment, prevention and / or amelioration of the most bothersome symptoms associated with migraine headache.

[0211] Administration In one embodiment of the present invention, the anti-CGRP antibodies, or CGRP-binding fragments thereof, as well as combinations of said antibodies or antibody fragments, described herein are administered to the subject at a concentration of about 0.1-100.0 mg / kg of the recipient subject's body weight. In a preferred embodiment of the present invention, the anti-CGRP antibodies, or CGRP-binding fragments thereof, as well as combinations of said antibodies or antibody fragments, described herein are administered to the subject at a concentration of about 0.4 mg / kg of the recipient subject's body weight and / or at a dose of 100 or 300 mg. In a preferred embodiment of the present invention, the anti-CGRP antibodies, or CGRP-binding fragments thereof, as well as combinations of said antibodies or antibody fragments, described herein are administered to the recipient subject at a frequency of once every 26 weeks or 6 months or less, once every 16 weeks or 4 months or less, once every 8 weeks or 2 months or less, once every 4 weeks or 1 month or less, once every 2 weeks or 6 months or less, once every week or less, or once every day or less. Generally, administration of successive doses may vary by plus or minus several days from the aforementioned schedule, for example, administration every three months or every twelve weeks would involve administration of doses that vary by plus or minus 1, 2, 3, 4, 5, 5, or 7 days from the scheduled dates.

[0212] The Fab fragment may be administered every two weeks or less, every week or less, every day or less, multiple times per day, and / or every few hours. In one embodiment of the invention, a patient receives 1 mg / kg to 40 mg / kg / day of the Fab fragment, in divided doses 1 to 6 times per day, or in sustained release form, effective to obtain the desired result.

[0213] It should be understood that the concentration of antibody or Fab administered to a given patient may be higher or lower than the exemplary administration concentrations listed above.

[0214] One of ordinary skill in the art will be able to determine effective dosages and frequency of administration by routine experimentation guided by, for example, the disclosure herein and the teachings in Goodman, LS, Gilman, A., Brunton, LL, Lazo, JS, & Parker, KL (2006). Goodman & Gilman's the pharmacological basis of therapeutics. New York: McGraw-Hill; Howland, RD, Mycek, MJ, Harvey, RA, Champe, PC, & Mycek, MJ (2006). Pharmacology. Lippincott's illustrated reviews. Philadelphia: Lippincott Williams & Wilkins; and Golan, DE (2008). Principles of pharmacology: the pathophysiologic basis of drug therapy. Philadelphia, Pa., [etc.]: Lippincott Williams & Wilkins.

[0215] In another embodiment of the invention, the anti-CGRP antibodies described herein, or CGRP-binding fragments thereof, as well as combinations of said antibodies or antibody fragments, are administered to a subject in a pharmaceutical formulation.

[0216] "Pharmaceutical composition" refers to a chemical or biological composition suitable for administration to a mammal. Such compositions may be specifically formulated for administration via one or more of a number of routes, including, but not limited to, buccal, extradermal, epidural, inhalation, intra-arterial, intracardiac, intraventricular, intradermal, intramuscular, intranasal, intraocular, intraperitoneal, intraspinal, intrathecal, intravenous, oral, parenteral, rectal by enema or suppository, subcutaneous, subdermal, sublingual, transdermal, and transmucosal, preferably intravenous. Additionally, administration may be by injection, powder, liquid, gel, drop, or other means of administration.

[0217] A "pharmaceutical excipient" or "pharmaceutical acceptable excipient" is a carrier, usually a liquid, in which an active therapeutic agent is formulated. In one embodiment of the present invention, the active therapeutic agent is a humanized antibody, or one or more fragments thereof, as described herein. The excipient generally does not provide any pharmacological activity to the formulation, but may provide chemical and / or biological stability, and release characteristics. Exemplary formulations are described, for example, in Remington's Pharmaceutical Sciences, 1999, 10:131-135, 19 ... th Ed., Grennaro, A., Ed., 1995 (incorporated by reference).

[0218] As used herein, "pharmaceutically acceptable carrier" or "excipient" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents that are physiologically compatible. In one embodiment, the carrier is suitable for parenteral administration. Alternatively, the carrier may be suitable for intravenous, intraperitoneal, intramuscular, or sublingual administration. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, their use in the pharmaceutical compositions of the present invention is contemplated. Supplementary active compounds can also be incorporated into the compositions.

[0219] Pharmaceutical compositions typically must be sterile and stable under the conditions of manufacture and storage. The present invention contemplates that the pharmaceutical composition is in lyophilized form. The composition may be formulated as a solution, microemulsion, liposome, or other ordered structure suitable for high drug concentration. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. The present invention further contemplates that a stabilizer is included in the pharmaceutical composition. Proper fluidity can be maintained, for example, by maintaining the required particle size in the case of dispersions and by the use of surfactants.

[0220] In many cases, it will be preferable to include an isotonic agent, such as sugars, polyalcohols, such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable composition can be achieved by including in the composition an agent that delays absorption, such as monostearate salts and gelatin. In addition, alkaline polypeptides can be formulated into time-release preparations, for example, in compositions that include slow-release polymers. The active compound can be prepared with a carrier that protects the compound from rapid release, such as controlled release preparations, including implants and microencapsulated delivery systems. Biodegradable biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid, and polylactic acid, polyglycolic acid copolymers (PLG). Many methods for the preparation of such preparations are known to those skilled in the art.

[0221] An exemplary composition comprises, consists essentially of, or consists of Ab6 in an aqueous solution, an excipient such as histidine, an isotonicity agent such as sorbitol, and a surfactant such as polysorbate 80. For example, the composition may comprise, consist essentially of, or consist of histidine (L-histidine), sorbitol, polysorbate 80, e.g., about 100 mg Ab6, about 3.1 mg L-histidine, about 40.5 mg sorbitol, and about 0.15 mg polysorbate 80 per mL volume, having a pH of about 5.8, or about the composition thereof, e.g., within 10% of these values, within 5% of these values, within 1% of these values, within 0.5% of these values, or within 0.1% of these values, and water. For example, the pH value may be within 10% of 5.8, i.e., 5.22 to 6.38. The Ab6 antibody comprises or consists of the variable light and heavy chain polypeptides of SEQ ID NO:222 and SEQ ID NO:202, respectively, or the variable light and heavy chain polypeptides of SEQ ID NO:221 and SEQ ID NO:201, respectively, or the variable light and heavy chain polypeptides of SEQ ID NO:221 and SEQ ID NO:566, respectively. The composition may be in the form of an aqueous solution or a concentrate (e.g., lyophilized) that, when reconstituted, for example, by the addition of water, results in the aforementioned configuration. An exemplary composition consists of 100 mg of the light and heavy chain polypeptides of SEQ ID NO:221 and SEQ ID NO:201, respectively, about 3.1 mg L-histidine, about 40.5 mg sorbitol, and about 0.15 mg polysorbate 80, and water QS, per mL, or about that configuration, for example, within 10% of these amounts, 5% of these amounts, within 1% of these amounts, within 0.5% of these amounts, or within 0.1% of these amounts. Another exemplary composition consists of 100 mg of light and heavy chain polypeptides of SEQ ID NO:221 and SEQ ID NO:566, respectively, about 3.1 mg L-histidine, about 40.5 mg sorbitol, and about 0.15 mg polysorbate 80, and water QS, or about the composition thereof, e.g., within 10% of these amounts, 5% of these amounts, within 1% of these amounts, within 0.5% of these amounts, or within 0.1% of these amounts. The composition may be suitable for intravenous or subcutaneous administration, preferably intravenous administration.For example, the composition may be suitable for mixing with intravenous solution (e.g., 0.9% sodium chloride) in an amount of about 100 mg to about 300 mg of antibody added to 100 mL of intravenous solution. Preferably, the composition may be shelf-stable for at least 1, 3, 6, 12, 18, or 24 months, e.g., exhibiting no more than 5% or no more than 10% aggregate formation of the antibody or fragment after storage at room temperature or when refrigerated at 4° C. for a specified period of time, or in an accelerated degradation test simulating storage for that period of time.

[0222] For each of the listed embodiments, the compound can be administered by various dosage forms.Any biologically acceptable dosage form known to those skilled in the art and their combinations are contemplated.Examples of such dosage forms include, but are not limited to, reconstitutable powder, elixir, solution, suspension, emulsion, powder, granule, particle, microparticle, dispersible granule, cachet, inhalant, aerosol inhalant, patch, particle inhalant, implant, depot implant, injection (including subcutaneous, intramuscular, intravenous, and intradermal, preferably intravenous), infusion, and combinations thereof.

[0223] The above description of various exemplary embodiments of the present invention is not intended to be exhaustive or to limit the invention to the precise form disclosed. For illustrative purposes, specific embodiments and examples of the present invention are described herein, but various equivalent modifications are possible within the scope of the present invention, as those skilled in the art will recognize. The teachings of the present invention provided herein can be applied to other purposes other than the examples described above.

[0224] These and other changes can be made to the invention in light of the above detailed description. In general, the terms used in the following claims should not be construed to limit the invention to the specific embodiments disclosed in the specification and the claims. Therefore, the invention is not limited by this disclosure; instead, the scope of the invention should be determined entirely by the following claims.

[0225] The invention may be practiced otherwise than as particularly described in the foregoing description and examples Numerous modifications and variations of the present invention are possible in light of the above teachings and, therefore, are within the scope of the appended claims.

[0226] Certain CGRP antibody polynucleotides and polypeptides are disclosed in the Sequence Listing accompanying this patent application, the disclosure of which is incorporated herein by reference in its entirety.

[0227] The entire disclosure of each document cited in the "Background," "Description of the Invention," and "Examples" sections (including patents, patent applications, journal articles, abstracts, manuals, books, or other disclosures) is hereby incorporated by reference in its entirety.

[0228] The following examples are set forth to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what is regarded as the invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, concentrations, etc.), but some experimental error and deviation should be allowed for. Unless otherwise specified, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0229] Additional Exemplary Embodiments Further exemplary embodiments of the present invention are as follows.

[0230] S1. Use of an anti-CGRP antibody for the manufacture of a medicament for treating a most bothersome symptom (MBS) associated with migraine, comprising administering an anti-CGRP antibody to a migraine patient.

[0231] S2. Use of an anti-CGRP antibody for the manufacture of a medicament for treating a most bothersome symptom (MBS) associated with migraine, the use comprising administering an anti-CGRP antibody to a migraine patient, the migraine patient suffering from chronic migraine.

[0232] S3. Use of an anti-CGRP antibody for the manufacture of a medicament for treating a most bothersome symptom (MBS) associated with migraine, the use comprising administering an anti-CGRP antibody to a migraine patient, the patient suffering from episodic migraine headaches.

[0233] S4. The use of an anti-CGRP antibody of any one of embodiments S1-S3, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved 1-12 hours after completion of the administration or infusion, e.g., 1-5 hours after completion of the administration or infusion, 1-2 hours after completion of the administration or infusion, or about 2 hours after completion of the administration or infusion.

[0234] S5. The use of an anti-CGRP antibody of any one of embodiments S1 to S4, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within one month of the first administration of the anti-CGRP antibody.

[0235] S6. The use of an anti-CGRP antibody of any one of embodiments S1 to S5, wherein the patient's migraine-associated most bothersome symptoms (MBS) improve within 3 months of the first administration of the anti-CGRP antibody.

[0236] S7. The use of an anti-CGRP antibody of any one of embodiments S1 to S6, wherein the patient's migraine-associated most bothersome symptoms (MBS) improve within 6 months of the first administration of the anti-CGRP antibody.

[0237] S8. The use of an anti-CGRP antibody of any one of embodiments S1 to S7, wherein improvement persists for at least 3 months from the first administration of the anti-CGRP antibody.

[0238] S9. The use of an anti-CGRP antibody of any one of embodiments S1 to S8, wherein improvement persists for at least 6 months from the first administration of the anti-CGRP antibody.

[0239] S10. Use of an anti-CGRP antibody according to any one of embodiments S1 to S9, wherein the MBS is selected from the group consisting of: sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), aura, pain with activity, pain, pulsation / pulsation, cognitive confusion, fatigue, mood swings, sensitivity to smells (osmophobia or olfactophobia), visual effects, pressure / tension, pain (anatomical), eye pain, neck pain, dizziness, allodynia, inactivity, sensory disturbances, sleep disturbances and speech disorders.

[0240] S11. Use of an anti-CGRP antibody according to any one of embodiments S1 to S10, wherein the MBS is selected from the group consisting of sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), pain with activity, pain, pulsation / pulsation, cognitive confusion, fatigue, mood swings and sensitivity to smells (osmophobia or olfactophobia).

[0241] S12. Use of an anti-CGRP antibody for the manufacture of a medicament for improving a patient's global impression of migraine-related changes (PGIC), comprising administering an anti-CGRP antibody to a migraine patient.

[0242] S13. Use of an anti-CGRP antibody for the manufacture of a medicament for improving Patient Global Impression of Changes (PGIC) associated with migraine, the use comprising administering an anti-CGRP antibody to a migraine patient, the migraine patient suffering from chronic migraine.

[0243] S14. Use of an anti-CGRP antibody for the manufacture of a medicament for improving a Patient's Global Impression of Changes (PGIC) associated with migraine, the use comprising administering an anti-CGRP antibody to a migraine patient, the patient suffering from episodic migraine.

[0244] S15. The use of an anti-CGRP antibody according to any one of embodiments S12 to S14, wherein administration of the medicament improves Patient Global Impression of Changes (PGIC) associated with migraine headache within one month of the first administration of the anti-CGRP antibody.

[0245] S16. The use of an anti-CGRP antibody according to any one of embodiments S12 to S14, wherein administration of the medicament improves Patient Global Impression of Changes (PGIC) associated with migraine headaches within 3 months of the first administration of the anti-CGRP antibody.

[0246] S17. The use of an anti-CGRP antibody according to any one of embodiments S12 to S14, wherein administration of the medicament improves Patient Global Impression of Changes (PGIC) associated with migraine headaches within 6 months of the first administration of the anti-CGRP antibody.

[0247] S18. The use of an anti-CGRP antibody according to any one of embodiments S12 to S17, wherein administration of the medicament improves Patient Global Impression of Changes (PGIC) associated with migraine, and this improvement persists for at least 3 months after the first administration of the anti-CGRP antibody.

[0248] S19. The use of an anti-CGRP antibody according to any one of embodiments S12 to S18, wherein administration of the medicament improves Patient Global Impression of Changes (PGIC) associated with migraine, and this improvement persists for at least 6 months after the first administration of the anti-CGRP antibody.

[0249] S20. The use of an anti-CGRP antibody according to any one of embodiments S1 to S19, wherein the medicament is for intravenous or subcutaneous injection.

[0250] S21. The use of an anti-CGRP antibody according to any one of embodiments S1 to S20, wherein the medicament is for intravenous infusion.

[0251] S22. The use of an anti-CGRP antibody according to any one of embodiments S1 to S21, wherein the patient is headache-free 2 hours after completion of the administration or infusion.

[0252] S23. The use of an anti-CGRP antibody of any one of embodiments S1 to S22, wherein the anti-CGRP antibody comprises Ab6.

[0253] S24. Use of an anti-CGRP antibody according to any one of embodiments S1 to S23, wherein the anti-CGRP antibody comprises light chain complementarity determining regions (CDRs) 1, 2, and 3 polypeptide sequences of SEQ ID NO: 224; SEQ ID NO: 226; and SEQ ID NO: 228, respectively.

[0254] S25. Use of an anti-CGRP antibody according to any one of embodiments S1 to S24, wherein the anti-CGRP antibody comprises light chain CDRs 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively.

[0255] S26. Use of an anti-CGRP antibody according to any one of embodiments S1 to S25, wherein the anti-CGRP antibody comprises heavy chain CDRs 1, 2, and 3 polypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 206; and SEQ ID NO: 208, respectively.

[0256] S27. Use of an anti-CGRP antibody according to any one of embodiments S1 to S26, wherein the anti-CGRP antibody comprises heavy chain CDRs 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively.

[0257] S28. Use of an anti-CGRP antibody according to any one of embodiments S1 to S27, wherein the anti-CGRP antibody comprises light chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, respectively.

[0258] S29. Use of an anti-CGRP antibody according to any one of embodiments S1 to S28, wherein the anti-CGRP antibody comprises light chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively.

[0259] S30. Use of an anti-CGRP antibody according to any one of embodiments S1 to S29, wherein the anti-CGRP antibody comprises a variable light chain polypeptide of SEQ ID NO: 222.

[0260] S31. Use of an anti-CGRP antibody according to any one of embodiments S1 to S30, wherein the anti-CGRP antibody comprises a variable light chain polypeptide encoded by SEQ ID NO: 232.

[0261] S32. Use of an anti-CGRP antibody according to any one of embodiments S1 to S31, wherein the anti-CGRP antibody comprises a variable heavy chain polypeptide of SEQ ID NO: 202.

[0262] S33. Use of an anti-CGRP antibody according to any one of embodiments S1 to S32, wherein the anti-CGRP antibody comprises a variable heavy chain polypeptide encoded by SEQ ID NO: 212.

[0263] S34. Use of an anti-CGRP antibody according to any one of embodiments S1 to S33, wherein the anti-CGRP antibody comprises a variable light chain polypeptide of SEQ ID NO: 222 and a variable heavy chain polypeptide of SEQ ID NO: 202.

[0264] S35. Use of an anti-CGRP antibody according to any one of embodiments S1 to S34, wherein the anti-CGRP antibody comprises a variable light chain polypeptide encoded by SEQ ID NO: 232 and a variable heavy chain polypeptide encoded by SEQ ID NO: 212.

[0265] S36. Use of an anti-CGRP antibody according to any one of embodiments S1 to S35, wherein the anti-CGRP antibody comprises a light chain polypeptide of SEQ ID NO: 221.

[0266] S37. Use of an anti-CGRP antibody according to any one of embodiments S1 to S36, wherein the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO: 231.

[0267] S38. Use of an anti-CGRP antibody according to any one of embodiments S1 to S37, wherein the anti-CGRP antibody comprises a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[0268] S39. Use of an anti-CGRP antibody according to any one of embodiments S1 to S38, wherein the anti-CGRP antibody comprises a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0269] S40. Use of an anti-CGRP antibody according to any one of embodiments S1 to S39, wherein the anti-CGRP antibody comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[0270] S41. Use of an anti-CGRP antibody according to any one of embodiments S1 to S40, wherein the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO: 231 and a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0271] S42. The use of an anti-CGRP antibody according to any one of embodiments S1 to S41, wherein the dosage of the anti-CGRP antibody is from about 100 mg to about 300 mg, or is about 100 mg, or is about 300 mg.

[0272] S43. The use of an anti-CGRP antibody according to any one of embodiments S1 to S42, wherein the dosage of the anti-CGRP antibody is 100 mg.

[0273] S44. The use of an anti-CGRP antibody according to any one of embodiments S1 to S43, wherein the medicament is for intravenous administration every 10 to 14 weeks, preferably every 11 to 13 weeks, more preferably every 12 weeks, at a dose of 100 mg of the anti-CGRP antibody.

[0274] S45. The use of an anti-CGRP antibody according to any one of embodiments S1 to S42, wherein the medicament is for intravenous administration every 10 to 14 weeks, preferably every 11 to 13 weeks, more preferably every 12 weeks, at a dose of 300 mg of the anti-CGRP antibody.

[0275] S46. The use of an anti-CGRP antibody according to any one of embodiments S1 to S45, wherein prior to administration of the medicament, the patient exhibits 1 to 10 migraine attacks per month in the month or 3 months prior to administration.

[0276] S47. The use of an anti-CGRP antibody according to any one of embodiments S1 to S46, wherein prior to administration of the medicament, the patient exhibits 2 to 8 migraine attacks per month in the month or 3 months prior to administration.

[0277] S48. The use of an anti-CGRP antibody according to any one of embodiments S1 to S47, wherein prior to administration of the medicament, the patient exhibits 3 to 7 migraine attacks per month in the month or 3 months prior to administration.

[0278] S49. The use of an anti-CGRP antibody according to any one of embodiments S1 to S48, wherein prior to administration of the medicament, the patient exhibits fewer than 25 headache days per month in the month or 3 months prior to administration.

[0279] S50. The use of an anti-CGRP antibody according to any one of embodiments S1 to S49, wherein prior to administration of the medicament, the patient exhibits fewer than 20 headache days per month in the month or 3 months prior to administration.

[0280] S51. The use of an anti-CGRP antibody according to any one of embodiments S1 to S50, wherein prior to administration of the medicament, the patient exhibits fewer than 15 headache days per month in the month or 3 months prior to administration.

[0281] S52. The use of an anti-CGRP antibody according to any one of embodiments S1 to S51, wherein prior to administration of the medicament, the patient exhibits less than 10 headache days per month in the month or 3 months prior to administration.

[0282] S53. The use of an anti-CGRP antibody according to any one of embodiments S1 to S52, wherein the patient was diagnosed with migraine headache at least 10 years prior to administration of the medicament.

[0283] S54. The use of an anti-CGRP antibody according to any one of embodiments S1 to S53, wherein the patient was diagnosed with migraine headache at least 15 years prior to administration of the medicament.

[0284] S55. The use of an anti-CGRP antibody according to any one of embodiments S1 to S54, wherein the patient was diagnosed with migraine at least 18 or at least 19 years prior to administration of the medicament.

[0285] S56. The use of an anti-CGRP antibody of any one of embodiments S1 to S55, wherein the patient further has at least a 50% reduction in the number of migraine days during the month following administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the medicament.

[0286] S57. The use of an anti-CGRP antibody of any one of embodiments S1 to S56, wherein the patient further has at least a 75% reduction in the number of migraine days during the month following administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the medicament.

[0287] S58. The use of an anti-CGRP antibody of any one of embodiments S1 to S57, wherein the patient further has a 100% reduction in the number of migraine days in the one month period following administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the medicament.

[0288] S59. The use of an anti-CGRP antibody of any one of embodiments S1 to S58, wherein the patient further has at least a 50% reduction in the number of migraine days in the 12 weeks after administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the medicament.

[0289] S60. The use of an anti-CGRP antibody of any one of embodiments S1 to S59, wherein the patient further has at least a 75% reduction in the number of migraine days in the 12 weeks after administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the medicament.

[0290] S61. The use of an anti-CGRP antibody of any one of embodiments S1 to S60, wherein the patient further has a 100% reduction in the number of migraine days in the 12 weeks after administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the medicament.

[0291] S62. The use of an anti-CGRP antibody according to any one of embodiments S1 to S61, wherein the medicament is further for administration of a second dose of the anti-CGRP antibody about 10 to 14 weeks, preferably 11 to 13 weeks, more preferably about 12 weeks or about 3 months after administration of the medicament.

[0292] S63. The use of an anti-CGRP antibody according to any one of embodiments S1 to S62, wherein the medicament comprises about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg of the anti-CGRP antibody.

[0293] S64. Use of an anti-CGRP antibody according to any one of embodiments S1 to S63, wherein the anti-CGRP antibody is non-glycosylated or, if glycosylated, contains only mannose residues.

[0294] S65. Use of an anti-CGRP antibody according to any one of embodiments S1 to S64, wherein the anti-CGRP antibody consists of a light chain polypeptide encoded by SEQ ID NO: 221 and a heavy chain polypeptide encoded by SEQ ID NO: 201 or SEQ ID NO: 566.

[0295] S66. Use of an anti-CGRP antibody according to any one of embodiments S1 to S65, wherein the anti-CGRP antibody consists of a light chain polypeptide encoded by SEQ ID NO: 231 and a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0296] S67. The use of an anti-CGRP antibody according to any one of embodiments S1 to S56, wherein the headache or migraine is diagnosed according to the International Classification of Headache Disorders, 3rd edition.

[0297] S68. Use of an anti-CGRP antibody according to any one of embodiments S1 to S67, wherein said anti-CGRP antibody is expressed in or obtained by Pichia pastoris.

[0298] S69. Use of an anti-CGRP antibody according to any one of embodiments S1 to S67, wherein said anti-CGRP antibody is expressed in or obtained by a CHO cell.

[0299] S70. Use of an anti-CGRP antibody according to any one of embodiments S1 to S69, wherein the anti-CGRP antibody or anti-CGRP antibody fragment is contained in a formulation comprising or consisting of histidine (L-histidine), sorbitol, polysorbate 80, and water.

[0300] S71. The use of an anti-CGRP antibody according to embodiment S70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 10% of said values, and has a pH of 5.8, or has amounts within + / - 10% of said values.

[0301] S72. The use of an anti-CGRP antibody according to embodiment S70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 5% of said values, and / or has a pH of 5.8, or has amounts within + / - 5% of said values.

[0302] S73. The use of an anti-CGRP antibody according to embodiment S70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 1% of said values, and / or has a pH of 5.8, or has amounts within + / - 1% of said values.

[0303] S74. The use of an anti-CGRP antibody according to embodiment S70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 0.5% of said values, and / or has a pH of 5.8, or has amounts within + / - 0.5% of said values.

[0304] S75. The use of an anti-CGRP antibody according to embodiment S70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 0.1% of said values, and / or has a pH of 5.8, or has amounts within + / - 0.1% of said values.

[0305] S76. Use of an anti-CGRP antibody according to any one of embodiments S1 to S75, wherein the anti-CGRP antibody has a dissociation constant of 10 pM or less, such as between 2 and 8 pM, such as between 3 and 6 pM, such as about 5 pM or less.

[0306] Further exemplary embodiments Further exemplary embodiments of the present invention are as follows.

[0307] E1. An anti-CGRP antibody for use in the treatment of most bothersome symptoms associated with migraine (MBS) in a patient suffering from migraine.

[0308] E2. The anti-CGRP antibody for use of embodiment E1, wherein the patient is suffering from chronic migraine headaches.

[0309] E3. The anti-CGRP antibody for use according to embodiment E1, wherein the patient is suffering from episodic migraine headaches.

[0310] E4. The anti-CGRP antibody for use of any one of embodiments E1-E3, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved 1 to 12 hours after completion of the administration or infusion, e.g., 1 to 5 hours after completion of the administration or infusion, 1 to 2 hours after completion of the administration or infusion, or about 2 hours after completion of the administration or infusion.

[0311] E5. The anti-CGRP antibody for use of any one of embodiments E1-E4, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within 1 month of the first administration of the anti-CGRP antibody.

[0312] E6. The anti-CGRP antibody for use of any one of embodiments E1-E5, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within 3 months of the first administration of the anti-CGRP antibody.

[0313] E7. The anti-CGRP antibody for use of any one of embodiments E1-E6, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within 6 months of the first administration of the anti-CGRP antibody.

[0314] E8. The anti-CGRP antibody for use of any one of embodiments E1-E7, wherein improvement persists for at least 3 months from the first administration of the anti-CGRP antibody.

[0315] E9. The anti-CGRP antibody for use of any one of embodiments E1-E8, wherein improvement persists for at least 6 months from the first administration of the anti-CGRP antibody.

[0316] E10. The anti-CGRP antibody for use according to any one of embodiments E1-E9, wherein the MBS is selected from the group consisting of: sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), aura, pain with activity, pain, pulsation / pulsation, cognitive confusion, fatigue, mood swings, sensitivity to smells (osmophobia or olfactophobia), visual effects, pressure / tension, pain (anatomical), eye pain, neck pain, dizziness, allodynia, inactivity, sensory disturbances, sleep disturbances and speech disturbances.

[0317] E11. An anti-CGRP antibody for use according to any one of embodiments E1 to E10, wherein the MBS is selected from the group consisting of sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), aura, pain with activity, pain, pulsation / pulsation, cognitive confusion, fatigue, mood swings and sensitivity to smells (osmophobia or olfactophobia).

[0318] E12. An anti-CGRP antibody for use in improving Patient Global Impression of Migraine-related Changes (PGIC) in a patient suffering from migraine.

[0319] E13. The anti-CGRP antibody for use according to embodiment E12, wherein the patient suffers from chronic migraine.

[0320] E14. The anti-CGRP antibody for use according to embodiment E12, wherein the patient suffers from episodic migraines.

[0321] E15. The anti-CGRP antibody for use according to any one of embodiments E12-E14, wherein an improvement in Patient Global Impression of Migraine-related Changes (PGIC) is observed within one month of the first administration of said anti-CGRP antibody.

[0322] E16. The anti-CGRP antibody for use according to any one of embodiments E12-E14, wherein an improvement in Patient Global Impression of Migraine-related Changes (PGIC) is observed within 3 months of the first administration of said anti-CGRP antibody.

[0323] E17. The anti-CGRP antibody for use according to any one of embodiments E12-E14, wherein an improvement in Patient Global Impression of Migraine-related Changes (PGIC) is observed within 6 months of the first administration of the anti-CGRP antibody.

[0324] E18. The anti-CGRP antibody for use of any one of embodiments E12-E17, wherein the improvement in Patient Global Impression of Migraine-related Changes (PGIC) persists for 3 months from the first administration of the anti-CGRP antibody.

[0325] E19. The anti-CGRP antibody for use of any one of embodiments E12-E18, wherein the improvement in Patient Global Impression of Migraine-related Changes (PGIC) persists for 6 months from the first administration of the anti-CGRP antibody.

[0326] E20. The anti-CGRP antibody for use of embodiments E1-E19, wherein the anti-CGRP antibody is for intravenous or subcutaneous infusion.

[0327] E21. The anti-CGRP antibody for use according to any one of embodiments E1-E20, wherein the anti-CGRP antibody is for intravenous infusion.

[0328] E22. The anti-CGRP antibody for use of any one of embodiments E1-E21, wherein the patient is headache-free 2 hours after completion of the administration or infusion.

[0329] E23. The anti-CGRP antibody for use of any one of embodiments E1-E22, wherein the anti-CGRP antibody comprises Ab6.

[0330] E24. The anti-CGRP antibody for use of any one of embodiments E1-E23, wherein the anti-CGRP antibody comprises light chain complementarity determining region (CDR) 1, 2, and 3 polypeptide sequences of SEQ ID NO: 224; SEQ ID NO: 226; and SEQ ID NO: 228, respectively.

[0331] E25. The anti-CGRP antibody for use of any one of embodiments E1-E24, wherein the anti-CGRP antibody comprises light chain CDRs 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively.

[0332] E26. The anti-CGRP antibody for use according to any one of embodiments E1-E25, wherein the anti-CGRP antibody comprises heavy chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 206; and SEQ ID NO: 208, respectively.

[0333] E27. The anti-CGRP antibody for use according to any one of embodiments E1-E26, wherein the anti-CGRP antibody comprises heavy chain CDRs 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively.

[0334] E28. The anti-CGRP antibody for use according to any one of embodiments E1-E27, wherein the anti-CGRP antibody comprises light chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, respectively.

[0335] E29. The anti-CGRP antibody for use according to any one of embodiments E1-E28, wherein the anti-CGRP antibody comprises light chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively.

[0336] E30. The anti-CGRP antibody for use of any one of embodiments E1-E29, wherein the anti-CGRP antibody comprises a variable light chain polypeptide of SEQ ID NO: 222.

[0337] E31. The anti-CGRP antibody for use of any one of embodiments E1-E30, wherein the anti-CGRP antibody comprises a variable light chain polypeptide encoded by SEQ ID NO: 232.

[0338] E32. The anti-CGRP antibody for use of any one of embodiments E1-E31, wherein the anti-CGRP antibody comprises a variable heavy chain polypeptide of SEQ ID NO: 202.

[0339] E33. The anti-CGRP antibody for use of any one of embodiments E1-E32, wherein the anti-CGRP antibody comprises a variable heavy chain polypeptide encoded by SEQ ID NO: 212.

[0340] E34. The anti-CGRP antibody for use of any one of embodiments E1-E33, wherein the anti-CGRP antibody comprises a variable light chain polypeptide of SEQ ID NO: 222 and a variable heavy chain polypeptide of SEQ ID NO: 202.

[0341] E35. The anti-CGRP antibody for use of any one of embodiments E1-E34, wherein the anti-CGRP antibody comprises a variable light chain polypeptide encoded by SEQ ID NO: 232 and a variable heavy chain polypeptide encoded by SEQ ID NO: 212.

[0342] E36. The anti-CGRP antibody for use of any one of embodiments E1-E35, wherein the anti-CGRP antibody comprises a light chain polypeptide of SEQ ID NO: 221.

[0343] E37. The anti-CGRP antibody for use of any one of embodiments E1-E36, wherein the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO: 231.

[0344] E38. The anti-CGRP antibody for use of any one of embodiments E1-E37, wherein the anti-CGRP antibody comprises a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[0345] E39. The anti-CGRP antibody for use of any one of embodiments E1-E38, wherein the anti-CGRP antibody comprises a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0346] E40. The anti-CGRP antibody for use of any one of embodiments E1-E39, wherein the anti-CGRP antibody comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[0347] E41. The anti-CGRP antibody for use of any one of embodiments E1-E40, wherein the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO: 231 and a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0348] E42. The anti-CGRP antibody for use according to any one of embodiments E1 to E41, wherein the dosage of the anti-CGRP antibody is from about 100 mg to about 300 mg, or is about 100 mg, or is about 300 mg.

[0349] E43. The anti-CGRP antibody for use of any one of embodiments E1-E42, wherein the dosage of the anti-CGRP antibody is 100 mg.

[0350] E44. The anti-CGRP antibody for use according to any one of embodiments E1-E43, wherein the anti-CGRP antibody is for intravenous administration at a dose of 100 mg of the anti-CGRP antibody every 10-14 weeks, preferably every 11-13 weeks, more preferably every 12 weeks.

[0351] E45. The anti-CGRP antibody for use according to any one of embodiments E1-E44, wherein the anti-CGRP antibody is for intravenous administration at a dosage of 300 mg of the anti-CGRP antibody administered every 10-14 weeks, preferably every 11-13 weeks, more preferably every 12 weeks.

[0352] E46. The anti-CGRP antibody for use according to any one of embodiments E1-E45, wherein prior to administration of said anti-CGRP antibody, the patient exhibits 1 to 10 migraine attacks per month in the month or 3 months prior to administration.

[0353] E47. The anti-CGRP antibody for use according to any one of embodiments E1-E46, wherein prior to administration of said anti-CGRP antibody, the patient exhibits 2 to 8 migraine attacks per month in the month or 3 months prior to administration.

[0354] E48. The anti-CGRP antibody for use according to any one of embodiments E1-E47, wherein prior to administration of said anti-CGRP antibody, the patient exhibits 3 to 7 migraine attacks per month in the month or 3 months prior to administration.

[0355] E49. The anti-CGRP antibody for use according to any one of embodiments E1-E48, wherein prior to administration of said anti-CGRP antibody, the patient exhibits fewer than 25 headache days per month in the month or 3 months prior to administration.

[0356] E50. The anti-CGRP antibody for use according to any one of embodiments E1-E49, wherein prior to administration of said anti-CGRP antibody, the patient exhibits fewer than 20 headache days per month in the month or 3 months prior to administration.

[0357] E51. The anti-CGRP antibody for use of any one of embodiments E1-E50, wherein prior to administration of said anti-CGRP antibody, the patient exhibits fewer than 15 headache days per month in the month or 3 months prior to administration.

[0358] E52. The anti-CGRP antibody for use of any one of embodiments E1-E51, wherein prior to administration of said anti-CGRP antibody, the patient exhibits fewer than 10 headache days per month in the month or 3 months prior to administration.

[0359] E53. The anti-CGRP antibody for use of any one of embodiments E1-E52, wherein the patient was diagnosed with migraine headache at least 10 years prior to administration of the anti-CGRP antibody.

[0360] E54. The anti-CGRP antibody for use of any one of embodiments E1-E53, wherein the patient was diagnosed with migraine at least 15 years prior to administration of the anti-CGRP antibody.

[0361] E55. The anti-CGRP antibody for use of any one of embodiments E1-E54, wherein the patient was diagnosed with migraine at least 18 or at least 19 years prior to administration of the anti-CGRP antibody.

[0362] E56. The anti-CGRP antibody for use of any one of embodiments E1-E55, wherein the patient has at least a 50% reduction in the number of migraine days during the month after administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the anti-CGRP antibody.

[0363] E57. The anti-CGRP antibody for use of any one of embodiments E1-E56, wherein the patient has at least a 75% reduction in the number of migraine days during the month after administration of the antibody compared to the baseline number of migraine days experienced by the patient prior to administration of the anti-CGRP antibody.

[0364] E58. The anti-CGRP antibody for use of any one of embodiments E1-E57, wherein the patient has a 100% reduction in migraine days over a one-month period after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to administration of the anti-CGRP antibody.

[0365] E59. The anti-CGRP antibody for use of any one of embodiments E1-E58, wherein the patient has at least a 50% reduction in the number of migraine days in the 12 weeks after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to administration of the anti-CGRP antibody.

[0366] E60. The anti-CGRP antibody for use of any one of embodiments E1-E59, wherein the patient has at least a 75% reduction in the number of migraine days in the 12 weeks after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to administration of the anti-CGRP antibody.

[0367] E61. The anti-CGRP antibody for use of any one of embodiments E1-E60, wherein the patient has a 100% reduction in migraine days in the 12 weeks after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to administration of the anti-CGRP antibody.

[0368] E62. The anti-CGRP antibody for use according to any one of embodiments E1 to E61, wherein said use comprises administering to said patient a second dose of said anti-CGRP antibody about 10 to 14 weeks, preferably 11 to 13 weeks, more preferably about 12 weeks or about 3 months after administration of said anti-CGRP antibody.

[0369] E63. The anti-CGRP antibody for use of any one of embodiments E1-E62, wherein said use comprises administering about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg of the anti-CGRP antibody.

[0370] E64. The anti-CGRP antibody for use of any one of embodiments E1-E63, wherein the anti-CGRP antibody is aglycosylated, or, if glycosylated, contains only mannose residues.

[0371] E65. The anti-CGRP antibody for use according to any one of embodiments E1 to E64, wherein the anti-CGRP antibody consists of a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[0372] E66. The anti-CGRP antibody for use according to any one of embodiments E1-E65, wherein the anti-CGRP antibody consists of a light chain polypeptide encoded by SEQ ID NO: 231 and a heavy chain polypeptide encoded by SEQ ID NO: 211 or SEQ ID NO: 567.

[0373] E67. The anti-CGRP antibody for use according to any one of embodiments E1 to E66, wherein the headache or migraine is diagnosed according to the International Classification of Headache Disorders, 3rd edition.

[0374] E68. The anti-CGRP antibody for use according to any one of embodiments E1 to E67, wherein said anti-CGRP antibody is expressed in or obtained by Pichia pastoris.

[0375] E69. The anti-CGRP antibody for use according to any one of embodiments E1 to E68, wherein said anti-CGRP antibody is expressed in or obtained by a CHO cell.

[0376] E70. The anti-CGRP antibody for use of any one of embodiments E1-E69, wherein the anti-CGRP antibody or anti-CGRP antibody fragment is in a formulation comprising or consisting of histidine (L-histidine), sorbitol, polysorbate 80, and water.

[0377] E71. The anti-CGRP antibody for use according to embodiment E70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 10% of said values, and has a pH of 5.8, or has amounts within + / - 10% of said values.

[0378] E72. The anti-CGRP antibody for use according to embodiment E70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 5% of said values, and / or has a pH of 5.8, or has amounts within + / - 5% of said values.

[0379] E73. The anti-CGRP antibody for use according to embodiment E70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 1% of said values, and / or has a pH of 5.8, or has amounts within + / - 1% of said values.

[0380] E74. The anti-CGRP antibody for use according to embodiment E70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 0.5% of said values, and / or has a pH of 5.8, or has amounts within + / - 0.5% of said values.

[0381] E75. The anti-CGRP antibody for use according to embodiment E70, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 0.1% of said values, and / or has a pH of 5.8, or has amounts within + / - 0.1% of said values.

[0382] E76. An anti-CGRP antibody for use according to any one of embodiments E1-E75, wherein the anti-CGRP antibody has a dissociation constant of 10 pM or less, such as between 2 and 8 pM, such as between 3 and 6 pM, such as about 5 pM or less. The present invention may include the following aspects. [1] A method of ameliorating the most bothersome symptoms associated with migraine (MBS), comprising administering to a patient in need thereof an effective amount of an anti-CGRP antibody. [2] 10. The method of claim 1, wherein the patient suffers from chronic migraine headaches. [3] 10. The method of claim 1, wherein the patient suffers from episodic migraine headaches. [4] 10. The method of any one of the preceding claims, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved 1 to 12 hours after completion of the administration or infusion, such as 1 to 5 hours after completion of the administration or infusion, such as 1 to 2 hours after completion of the administration or infusion, or about 2 hours after completion of the administration or infusion. [5] 2. The method of any one of the preceding claims, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within one month of the first administration of the anti-CGRP antibody. [6] 2. The method of any one of the preceding claims, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within 3 months of the first administration of the anti-CGRP antibody. [7] 2. The method of any one of the preceding claims, wherein the patient's migraine-associated most bothersome symptoms (MBS) are improved within 6 months of the first administration of the anti-CGRP antibody. [8] 2. The method of any one of the preceding claims, wherein the improvement persists for at least 3 months from the first administration of the anti-CGRP antibody. [9] 2. The method of any one of the preceding claims, wherein the improvement persists for at least 6 months from the first administration of the anti-CGRP antibody.

[10] 10. The method of any one of the preceding claims, wherein the MBS is selected from the group consisting of sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), aura, pain with activity, pain, pulsation / pulsation, cognitive confusion, fatigue, mood swings, sensitivity to smells (osmophobia or olfactophobia), visual effects, pressure / tension, pain (anatomical), eye pain, neck pain, dizziness, allodynia, inactivity, sensory disturbances, sleep disturbances and speech disturbances.

[11] 10. The method of any one of the preceding claims, wherein the MBS is selected from the group consisting of sensitivity to light (photophobia), nausea / vomiting, headache, sensitivity to sound (phonophobia), aura, pain with activity, pain, pulsation / pulsation, cognitive confusion, fatigue, mood swings and sensitivity to smells (osmophobia or olfactophobia).

[12] 1. A method of improving Patient Global Impression of Changes associated with migraine (PGIC) comprising administering to a patient in need thereof an effective amount of an anti-CGRP antibody.

[13] 13. The method of claim 12, wherein the patient suffers from chronic migraine headaches.

[14] 14. The method of claim 13, wherein the patient suffers from episodic migraine headaches.

[15] 15. The method of claim 12, wherein an improvement in Patient Global Impression of Migraine-related Changes (PGIC) is observed within one month of the first administration of the anti-CGRP antibody.

[16] 15. The method of claim 12, wherein an improvement in Patient Global Impression of Migraine-related Changes (PGIC) is observed within 3 months of the first administration of the anti-CGRP antibody.

[17] 15. The method of claim 12, wherein an improvement in Patient Global Impression of Migraine-related Changes (PGIC) is observed within 6 months of the first administration of the anti-CGRP antibody.

[18] 18. The method of any one of claims 12 to 17, wherein the improvement persists for at least 3 months from the first administration of the anti-CGRP antibody.

[19] 19. The method of any one of claims 12-18, wherein the improvement persists for at least 6 months from the first administration of the anti-CGRP antibody.

[20] 13. The method of any one of the preceding claims, wherein the administration is by injection, such as intravenous or subcutaneous injection.

[21] 10. The method of any one of the preceding claims, wherein the administration is by intravenous infusion.

[22] 10. The method of any one of the preceding claims, wherein the patient is headache-free 2 hours after completion of the administration or infusion.

[23] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises Ab6.

[24] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises light chain complementarity determining region (CDR) 1, 2, and 3 polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, respectively.

[25] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises light chain CDRs 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively.

[26] 20. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises the heavy chain CDRs 1, 2, and 3 polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, respectively.

[27] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises heavy chain CDRs 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively.

[28] 20. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises light chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:224; SEQ ID NO:226; and SEQ ID NO:228, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences of SEQ ID NO:204; SEQ ID NO:206; and SEQ ID NO:208, respectively.

[29] 20. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises light chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:234; SEQ ID NO:236; and SEQ ID NO:238, respectively, and heavy chain CDR 1, 2, and 3 polypeptide sequences encoded by SEQ ID NO:214; SEQ ID NO:216; and SEQ ID NO:218, respectively.

[30] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a variable light chain polypeptide of SEQ ID NO: 222.

[31] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a variable light chain polypeptide encoded by SEQ ID NO: 232.

[32] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a variable heavy chain polypeptide of SEQ ID NO: 202.

[33] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a variable heavy chain polypeptide encoded by SEQ ID NO:212.

[34] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a variable light chain polypeptide of SEQ ID NO: 222 and a variable heavy chain polypeptide of SEQ ID NO: 202.

[35] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a variable light chain polypeptide encoded by SEQ ID NO:232 and a variable heavy chain polypeptide encoded by SEQ ID NO:212.

[36] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a light chain polypeptide of SEQ ID NO: 221.

[37] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO: 231.

[38] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[39] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a heavy chain polypeptide encoded by SEQ ID NO:211 or SEQ ID NO:567.

[40] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[41] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO:231 and a heavy chain polypeptide encoded by SEQ ID NO:211 or SEQ ID NO:567.

[42] 2. The method of claim 1, wherein the dosage of the anti-CGRP antibody is from about 100 mg to about 300 mg, or is about 100 mg, or is about 300 mg.

[43] 2. The method of any one of the preceding claims, wherein the dosage of the anti-CGRP antibody is 100 mg.

[44] 13. The method of any one of the preceding claims, further comprising intravenously administering 100 mg of the anti-CGRP antibody every 10-14 weeks, preferably every 11-13 weeks, more preferably every 12 weeks.

[45] The method according to any one of claims 1 to 42, wherein the dose of the anti-CGRP antibody is 100 mg.

[46] 46. ​​The method of any one of claims 1 to 42 or 45, further comprising intravenously administering 300 mg of the anti-CGRP antibody every 10 to 14 weeks, preferably every 11 to 13 weeks, more preferably every 12 weeks.

[47] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits 1 to 10 migraine attacks per month in the month or 3 months prior to administration.

[48] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits 2 to 8 migraine attacks per month in the month or 3 months prior to administration.

[49] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits 3 to 7 migraine attacks per month in the month or 3 months prior to administration.

[50] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits fewer than 25 headache days per month in the month or 3 months prior to administration.

[51] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits fewer than 20 headache days per month in the month or 3 months prior to administration.

[52] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits fewer than 15 headache days per month in the month or 3 months prior to administration.

[53] 10. The method of any one of the preceding claims, wherein prior to said administration, the patient exhibits fewer than 10 headache days per month in the month or 3 months prior to administration.

[54] 2. The method of any one of the preceding claims, wherein the patient was diagnosed with migraine headache at least 10 years prior to said administering.

[55] 2. The method of any one of the preceding claims, wherein the patient was diagnosed with migraine headache at least 15 years prior to said administering.

[56] 2. The method of any one of the preceding claims, wherein the patient was diagnosed with migraine at least 18 or at least 19 years prior to said administering.

[57] 2. The method of any one of the preceding claims, wherein the patient has at least a 50% reduction in the number of migraine days over a one-month period after administering the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[58] 2. The method of any one of the preceding claims, wherein the patient has at least a 75% reduction in the number of migraine days over a one-month period after administering the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[59] 2. The method of any one of the preceding claims, wherein the patient has a 100% reduction in migraine days over a one-month period after administering the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[60] 2. The method of any one of the preceding claims, wherein the patient has at least a 50% reduction in the number of migraine days in the 12 week period after administering the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[61] 2. The method of any one of the preceding claims, wherein the patient has at least a 75% reduction in the number of migraine days in the 12 week period after administering the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[62] 2. The method of any one of the preceding claims, wherein the patient has a 100% reduction in migraine days in the 12 week period after administration of the antibody, compared to the baseline number of migraine days experienced by the patient prior to said administration.

[63] 2. The method of any one of the preceding claims, further comprising administering to the patient a second dose of the anti-CGRP antibody about 10 to 14 weeks, preferably 11 to 13 weeks, more preferably about 12 weeks or about 3 months after said administration.

[64] 2. The method of any one of the preceding claims, wherein said administering comprises administering about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, or about 300 mg of the anti-CGRP antibody.

[65] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody is aglycosylated, or, if glycosylated, contains only mannose residues.

[66] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody consists of a light chain polypeptide of SEQ ID NO: 221 and a heavy chain polypeptide of SEQ ID NO: 201 or SEQ ID NO: 566.

[67] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody consists of a light chain polypeptide encoded by SEQ ID NO:231 and a heavy chain polypeptide encoded by SEQ ID NO:211 or SEQ ID NO:567.

[68] 10. The method according to any one of the preceding claims, wherein the headache or migraine is diagnosed according to the International Classification of Headache Disorders, 3rd edition.

[69] 13. The method according to any one of the preceding claims, wherein the anti-CGRP antibody is expressed in Pichia pastoris or obtainable by expression in Pichia pastoris.

[70] 69. The method according to any one of claims 1 to 68, wherein the anti-CGRP antibody is expressed in or obtained by expression in a CHO cell.

[71] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody or anti-CGRP antibody fragment is in a formulation comprising or consisting of histidine (L-histidine), sorbitol, polysorbate 80, and water.

[72] 72. The method of claim 71, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 10% of said values, and has a pH of 5.8, or has a pH within + / - 10% of said values.

[73] 72. The method of claim 71, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 5% of said values, and / or has a pH of 5.8, or has a pH within + / - 5% of said values.

[74] 72. The method of claim 71, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 1% of said values, and / or has a pH of 5.8, or has a pH within + / - 1% of said values.

[75] 72. The method of claim 71, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 0.5% of said values, and / or has a pH of 5.8, or has a pH within + / - 0.5% of said values.

[76] 72. The method of claim 71, wherein the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg L-histidine, 40.5 mg sorbitol, and 0.15 mg polysorbate 80 per mL volume, or has amounts of each component within + / - 0.1% of said values, and / or has a pH of 5.8, or has a pH within + / - 0.1% of said values.

[77] 2. The method of any one of the preceding claims, wherein the anti-CGRP antibody has a dissociation constant of 10 pM or less, such as 2 to 8 pM, such as 3 to 6 pM, such as about 5 pM or less, optionally measured by surface plasmon resonance, optionally performed at 25°C or 37°C.

[78] 2. The method of any one of the preceding claims, which improves both MBS and PGIC in the patient. EXAMPLES

[0383] The following examples are provided to illustrate the invention, but should not be construed as limiting the scope of the claims in any way.

[0384] Example 1 Preparation of antibodies that bind to CGRP The preparation of an exemplary anti-CGRP antibody, Ab6, having the sequence of Figures 1-12 is disclosed in co-owned PCT application WO 2012 / 162243, published November 29, 2012, the contents of which are incorporated herein by reference. This application illustrates the synthesis of these antibodies in Pichia pastoris cells. This application further contemplates the synthesis of the anti-CGRP antibody, Ab6, specifically in CHO cells.

[0385] Example 2 A human clinical trial evaluating the safety and efficacy of anti-CGRP antibodies in patients with chronic migraine This example describes a randomized, double-blind, placebo-controlled clinical trial evaluating the safety and efficacy of Ab6 for chronic migraine prevention. In this study, 1,072 patients were randomized to receive Ab6 (300 mg or 100 mg) or placebo, administered by infusion once every 12 weeks. The study design is shown in Figure 13. To be eligible for the study, patients must experience at least 15 headache days per month, of which at least 8 days must meet the criteria for migraine. Patients who participated in the study had an average of 16.1 migraine days per month at baseline. The primary endpoint of the study was the change from baseline in mean monthly migraine days (MMD) over weeks 1-12 after the first infusion of Ab6. The change from baseline in mean monthly migraine days (MMD) after the second infusion at week 12 was also evaluated. The results are shown in Figure 14.

[0386] Study endpoints further included patient-identified MBS as part of the predefined key secondary endpoint. At screening, patients verbally identified their migraine-related MBS, which were pooled across treatment arms in this analysis. Changes from baseline in these symptoms were then assessed by patients during each month of the study, beginning on day 0.

[0387] In this study, patients verbally identified their most bothersome migraine-related symptoms (MBS) at screening. Patients' migraine-related MBS were then categorized by the investigator into a predefined list of eight symptoms or into an "other" option. The predefined list included the terms nausea, vomiting, sensitivity to light, sensitivity to sound, confusion, fatigue, pain with activity, and mood swings. The "other" option provided investigators with an open-ended opportunity to identify any migraine-related symptom that patients described as most bothersome but that did not easily fit into the checklist of symptoms included in the Work Study Checklist. For patients who selected the "other" category for their MBS, their "written" responses were subsequently recorded and recategorized into the predefined list or into a new symptom type. At subsequent visits, patients were asked to assess the change in their self-reported MBS from the screening visit on a 7-point scale as shown below.

[0388] [Table 1]

[0389] In addition to the MBS, patients were also asked to evaluate the effectiveness of the treatment in terms of the Patient Global Impression of Change (PGIC), a parameter that includes a single question assessing the patient's own impression of the overall change in their disease status since the start of the study. This parameter was also assessed by patients at the same time points during the study on the same 7-point scale used to evaluate the change in the MBS shown above. In Figures 16-22, the "worsening" category includes "minimally worsened," "significantly worsened," and "very significantly worsened."

[0390] At the screening visit, patients presented with a wide range of symptoms as their MBS, with the "other" category being the most common response (40%-42% across the three treatment groups). Patients who selected the "other" category generally provided more detailed information and / or had more than one symptom as their MBS, which resulted in these symptoms being recorded. A comprehensive list of MBS is summarized in Table 1 below.

[0391] [Table 2]

[0392] The most commonly reported symptoms were photophobia, nausea / vomiting, pain with activity, pain, headache, phonophobia, pulsation / pulsation, cognitive confusion, fatigue, mood swings, and sensitivity to smell, with at least 10 patients in each category reporting these events as their MBS. At the end of the 28-day screening period (i.e., before dosing at the baseline visit), patients were asked to rate the change in their identified MBS from very much worse to very much better, and >90% reported no change in their MBS. This is shown in Figure 16. This suggests that in this cohort with chronic migraine, there was little fluctuation in the bothersomeness of the patient-identified MBS during the screening period.

[0393] Infusions of Ab6 at doses of 100 mg and 300 mg significantly reduced mean MMD over months 1-3 of the study, with further reductions following an additional infusion in week 12 of the study, an effect shown in FIG.

[0394] Efficacy of Ab6 on MBS was demonstrated at 1 month (FIG. 17), 3 months (FIG. 19), and 6 months (FIG. 21) after the first infusion of Ab6 at doses of 100 mg and 300 mg. Efficacy of Ab6 on PGIC was demonstrated at 1 month (FIG. 18), 3 months (FIG. 20), and 6 months (FIG. 22) after the first infusion of Ab6 at doses of 100 mg and 300 mg. Efficacy on these parameters was sustained or increased over 6 months through two doses of Ab6. At 1 month, 75-82% of patients treated with Ab6 showed some level of improvement compared to 56-59% of patients treated with placebo. At 3 months, the assessment of improvement was similar to that at 1 month. At 6 months, approximately 80% of patients treated with Ab6 showed 1 or more category level improvement in MBS and PGIC. The distribution of ratings for improvement on the MBS and PGIC were similar at each time point, suggesting that two similarly rated scales move in parallel in patients with chronic migraine. These data suggest that improvement in patient-identified most bothersome symptoms related to migraine correlates highly with patient perception of improvement in disease state in patients with chronic migraine.

[0395] The administered antibody, Ab6, is an anti-CGRP antibody consisting of a light chain polypeptide of SEQ ID NO:221 and a heavy chain polypeptide of SEQ ID NO:201.

[0396] Patient characteristics are summarized in Table 2 below, with separate columns for patients receiving placebo, 100 mg antibody, or 300 mg antibody. The mean number of years since migraine diagnosis for patients ranged from 17.0 to 19.0 years, the mean duration of suffering from chronic migraine ranged from 11.5 to 12.4 years, and 44.3 to 45.2% of patients were utilizing at least one preventive medication. In addition, patients with a dual diagnosis of chronic migraine and medication overuse, excluding opioid and butalbital overuse, were included in the study. At baseline, the mean number of migraine days per month in both antibody treatment groups was 16.1 days, and in the placebo group, the mean number of migraine days per month was 16.2 days.

[0397]

Table 3

Claims

1. 1. A pharmaceutical composition for improving migraine associated most bothersome symptoms (MBS) in a patient comprising an anti-CGRP antibody, MBS is nausea or vomiting; the anti-CGRP antibody comprises a light chain polypeptide encoded by SEQ ID NO:231 and a heavy chain polypeptide encoded by SEQ ID NO:211 or SEQ ID NO:567; and the amount of the anti-CGRP antibody in the pharmaceutical composition is about 100 mg to about 300 mg, or is about 100 mg, or is about 300 mg; Pharmaceutical compositions.

2. The anti-CGRP antibody is contained in a formulation comprising or consisting of histidine (L-histidine), sorbitol, polysorbate 80, and water, wherein Optionally, the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 10% of said values, and has a pH of 5.8, or has a pH within + / - 10% of said values; Optionally, the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 5% of said values, and / or has a pH of 5.8, or has a pH within + / - 5% of said values, or Optionally, the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 1% of said values, and / or has a pH of 5.8, or has a pH within + / - 1% of said values, or Optionally, the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 0.5% of said values, and / or has a pH of 5.8, or has a pH within + / - 0.5% of said values, or Optionally, the formulation comprises or consists of 100 mg of anti-CGRP antibody, 3.1 mg of L-histidine, 40.5 mg of sorbitol, and 0.15 mg of polysorbate 80 per mL volume, or has amounts of each component within + / - 0.1% of said values, and / or has a pH of 5.8, or has a pH within + / - 0.1% of said values. The pharmaceutical composition of claim 1.

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