Treating pain

Abstract

The present invention is particularly directed to the treatment of pain. For example, chimeric Clostridial neurotoxins are provided for use in treating pain by inhibiting the release of pain mediators from neurons containing Aδ or C nerve fibers, wherein the chimeric Clostridial neurotoxins bind to neurons containing Aδ or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxins comprise a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C Methods, uses, kits, and unit dosage forms are also provided.

Description

[Technical Field]

[0001] The present invention relates to the treatment of disorders such as pain. [Background technology]

[0002] Pain is an unpleasant sensory and emotional experience associated with or resembling actual or potential tissue damage. Pain is also described as a neurological condition characterized by pathological changes in the nervous system, or more precisely, dysfunction of the endogenous nociceptive system (Raffaeli & Arnaudo (2017), J Pain Res, 10, 2003-2008).

[0003] Nociception is the process by which information about actual tissue damage (or the potential for such damage if noxious stimuli are continually presented) is relayed to the brain. The sensory neurons involved in nociception are classified into three main groups: group A, group B, and group C (Yam et al (2018), Int J Mol Sci, 19, 8, 2164).

[0004] Group A nerve fibers are classified as myelinated fibers and can be further subdivided into Aα, Aβ, Aγ, and Aδ, each with distinct properties. These fibers generally terminate in lamina I, III, IV, and V of the dorsal horn of the spinal cord, with some lamina II forming medial projections. Both Ia and Ib sensory fibers from muscle spindle endings and Golgi tendons are Aα type. Aβ fibers are generally low-threshold, cutaneous, slow- or fast-adapting mechanoreceptors, and contain type II afferents from stretch receptors. Aβ fibers generally reside in lamina III and IV. Aγ fibers may contain type II afferents from stretch receptors. Aδ fibers, along with type III afferents, may contain thermal and mechanical nociceptors that terminate in lamina I and V of the Rexed nerve. Aδ fibers are generally the least myelinated and can have relatively fast conduction velocities of approximately 30 m / s. Aδ fibers are generally about 2-5 μm in diameter and generally respond to brief tingling pain.

[0005] Group B nerve fibers are moderately myelinated and have a normal conduction velocity of 3-14 m / s. The preganglionic nerve fibers of the autonomic nervous system (ANS) and normal visceral afferent fibers belong to this group.

[0006] Group C nerve fibers are unmyelinated, generally less than 2 μm in diameter, and generally have a relatively slow conduction velocity of up to approximately 2 m / s. Nerve fibers of the dorsal root of the spinal cord (type IV afferent fibers) and postganglionic fibers of the ANS can be classified into this group. All of these fibers have a primary nociceptive function, carrying sensory information and collecting approximately 70% of the afferent nociceptive information, which then enters the spinal cord. C fibers may terminate in layers I and II of the gray matter of the spinal cord. In terms of nociception, C-fiber nociceptors may be polymodal, being activated by thermal, mechanical, and / or chemical stimuli. For example, C fibers can also be activated by poorly localized stimuli. In terms of neurochemistry, C fibers can be classified as either peptidergic or nonpeptidergic, and approximately 50% of these fibers express neuropeptides, including calcitonin gene-related peptide (CGRP), neurokinins, and substance P (SP).

[0007] Neurotransmitters involved in pain are diverse and include all major types of neurotransmitters, such as inflammatory mediators: prostaglandin E2 (PGE2), prostacyclin (PGI2), leukotriene B4 (LTB4), nerve growth factor (NGF), protons, bradykinin (BK), ATP, adenosine, SP, neurokinin A (NKA), neurokinin B (NKB), 5-hydroxytryptamine (5-HT), histamine, glutamate, norepinephrine (NE), and nitric oxide (NO); and non-inflammatory mediators: CGRP, gamma-aminobutyric acid (GABA), opioid peptides, glycine, and cannabinoids (Yam et al. (2018), Int J Mol Sci, 19, 8, 2164).

[0008] Of particular therapeutic interest is CGRP. CGRP is widely produced in both the central and peripheral nervous systems, but is primarily present in primary afferent nerves. As a direct derivative of the dorsal root ganglion (DRG), CGRP can be found in the dorsal horn of the spinal cord and may be involved in the conduction of noxious stimuli. CGRP is related to the excitatory action of SP, which is mediated by Ca 2+CGRP receptors (calcitonin receptor-like receptors (CALCRL)) are typically located in the nucleus accumbens, suggesting that the CNS may regulate CGRP-mediated pain transmission. CGRP is widely distributed throughout the peripheral and central nervous systems, and its receptors are expressed in pain pathways. CGRP-like immunoreactivity (CGRP-LI) is typically found in 40-50% of DRG neurons. Furthermore, CGRP typically coexists with other neuropeptides, such as substance P and neurokinins, in DRG neurons. Peripheral CGRP-LI fibers may terminate in lamina I, III, and V of the spinal cord, and CGRP-containing DRG neurons innervate joints. Thus, CGRP and its receptors may be widely distributed in peripheral and central pain pathways (Schou et al. (2017), The Journal of Headache and Pain, 18, 34, 1-17). In animals, CGRP can be released from peripheral and central nerve terminals in response to noxious pain and / or mechanical stimulation of the skin. In rats, the majority of circulating CGRP can be released from perivascular nerve terminals. Acute and chronic nociception can cause changes in CGRP release from sensory nerve terminals and central terminals to the dorsal horn of the spinal cord. CGRP is known to be one of the most potent vasodilators. Two isoforms, α-CGRP and β-CGRP, have been characterized (Russell et al. (2014), Physiol Rev, 94, 4, 1099-1142). Isoform α is primarily expressed in primary sensory neurons, while isoform β is found primarily in intrinsic enteric neurons. The mature form of this neuropeptide consists of 37 amino acids and is expressed primarily in sensory neurons of the DRG and trigeminal ganglion. The mature form is stored in vesicles localized in the terminal regions of central and peripheral nerve endings, from which it can be secreted into the dorsal spinal cord or various peripheral tissues, particularly into surrounding blood vessels, where it may regulate vascular tone. Furthermore, the presence of CGRP-positive nociceptor networks has been observed in meningeal vessels of rodents and humans, and approximately 40-50% of trigeminal ganglion neurons have been found to be positive for CGRP.Furthermore, CGRP expression has been observed in CNS regions such as the hypothalamus, thalamus, periaqueductal gray, superior and inferior colliculus, amygdala, trigeminocervical complex, and cerebellum. Given the ability of CGRP to alter synaptic and neuronal activity in the trigeminocervical complex and transmit nociceptive signals to the thalamus and cortical regions, these brain regions may be relevant to the pathophysiology of migraine (Tardiolo et al. (2019), Int J Mol Sci, 20(12), 2932).

[0009] Conventional treatments for pain (e.g., CGRP-associated pain) include monoclonal antibodies and small molecule antagonists that target pain mediators (e.g., CGRP) already released by presynaptic neurons. As an alternative approach, certain conventional therapeutic agents target the receptors for pain mediators. These approaches are associated with many disadvantages, including effects on systemic chemical mediators (e.g., CGRP); nausea; vomiting; gastrointestinal upset; diarrhea; bradycardia; hypotension; bronchospasm; dyspnea; fatigue; insomnia; dizziness; dry mouth; flushing; hot or cold sensations; chest pain; constipation; pruritus; drowsiness; tinnitus; restlessness; muscle spasms; injection site pain; upper respiratory tract infection; fatigue; nasopharyngitis; injection site erythema; injection site induration; anxiety; depression; injection site pruritus; influenza; urinary tract infection; somnolence; paresthesia; increased heart rate; stroke; and / or heart attack (Woo (2020), Nature, 586, S4-S6 and Tardiolo et al (2019), Int J Mol Sci, 20(12), 2932). Therefore, there is a need for improved pain treatments that have fewer side effects and / or block pain mediators at the point of release.

[0010] The present invention solves one or more of the problems set forth above. Summary of the Invention

[0011] The present inventors have demonstrated that, unlike BoNT / A, botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N domain), and the BoNT / B receptor binding domain (HC We have found that chimeric clostridial neurotoxins containing the clostridial domain (or clostridial domain) can bind to neurons containing Aδ or C nerve fibers that secrete pain mediators and can be effective in inhibiting the release of these mediators from these neurons. Thus, through such inhibition, the chimeric clostridial neurotoxins of the invention can function as analgesics capable of treating pain. In particular, we have shown that the claimed chimeric clostridial neurotoxins can be effective in inhibiting CGRP release from these neurons. Thus, through inhibition of CGRP release, the chimeric clostridial neurotoxins of the invention can function as analgesics capable of treating CGRP-associated pain.

[0012] Without wishing to be bound by theory, it is believed that the chimeric clostridial neurotoxins of the present invention may prevent pain mediators (e.g., CGRP) from reaching adjacent and distant cells by blocking the chemical mediator at the point of secretion. This advantageously allows selective blocking of pain-associated abnormal mediator release, sparing mediator release elsewhere; provides a therapeutic agent with a longer duration of action (fewer side effects and / or a wider margin of safety than non-chimeric clostridial neurotoxins); and / or reduces side effects compared to conventional therapies. In particular, blocking CGRP action and / or CGRP receptors once released by conventional therapeutic agents can cause nausea, fatigue, increased heart rate, stroke, and / or heart attack. These side effects can be minimized / avoided by the present invention.

[0013] The inventors have further found that chimeric Clostridial neurotoxins can cleave SNAP25 in central nervous system structures associated with migraine pathophysiology. Advantageously, chimeric Clostridial neurotoxins can be particularly effective in treating migraine headaches (e.g., migraine pain). DETAILED DESCRIPTION OF THE INVENTION

[0014] In one aspect, the present invention provides a chimeric clostridial neurotoxin for use in treating pain, wherein the chimeric clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0015] In one aspect, the present invention provides a method of treating pain, the method comprising administering to a subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0016] In one aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for the treatment of pain, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0017] In one aspect, the present invention provides a chimeric Clostridial neurotoxin for use in treating migraine headaches (preferably migraine pain), wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0018] In one aspect, the present invention provides a method of treating migraine (preferably migraine pain), the method comprising administering to a subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0019] In one aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for the treatment of migraine (preferably migraine pain), wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0020] Migraine may be episodic migraine or chronic migraine (preferably chronic migraine). If a subject experiences headache (e.g., migraine) less than 15 days per month (e.g., at least 1 day but less than 15 days per month), preferably if a subject experiences headache (e.g., migraine) at least 4 days but less than 15 days per month, the subject may suffer from episodic migraine. In other words, episodic migraine can be defined as headache (e.g., migraine) less than 15 days per month (e.g., at least 1 day but less than 15 days per month), preferably headache (e.g., migraine) at least 4 days but less than 15 days per month. If a subject experiences headache (e.g., migraine) at least 15 days per month, the subject may suffer from chronic migraine. If a subject experiences headache (e.g., migraine) at least 15 days per month for at least 3 months, and migraine characteristics are present for at least 8 days per month, the subject may suffer from chronic migraine. In other words, chronic migraine can be defined as headache (e.g., migraine) at least 15 days per month. In other words, chronic migraine may be defined as headaches (e.g., migraines) that are characterized by migraine headaches at least 8 days per month and occur at least 15 days per month for at least 3 months. In one embodiment, chronic migraines may last 4 hours or more per day. In addition to headaches, migraines may be accompanied by one or more additional symptoms, including increased light sensitivity, nausea, and / or vomiting.

[0021] Preferably, when treating migraine headaches, the chimeric Clostridial neurotoxin treats migraine pain.

[0022] In one aspect, the present invention provides a chimeric Clostridial neurotoxin for use in treating pain by inhibiting the release of pain mediators from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0023] In a related aspect, the present invention provides a method of treating pain by inhibiting the release of pain mediators from neurons comprising Aδ nerve fibers or C nerve fibers, the method comprising administering to a subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0024] In another related aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for treating pain by inhibiting the release of pain mediators from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0025] In one embodiment, the present invention provides a chimeric clostridial neurotoxin for use in treating CGRP-associated pain by inhibiting the release of CGRP from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C Corresponding methods of treatment and uses are also provided.

[0026] A mediator can be any molecule released from neurons that has a role in a disorder (such as pain), and inhibition of the release of said mediator by a chimeric Clostridial neurotoxin of the invention can treat said disorder (e.g., can treat pain).

[0027] The mediator may be a neurotransmitter.

[0028] The inhibition of mediator release from neurons may be partial or complete, preferably complete. For example, the chimeric Clostridial neurotoxin may inhibit at least 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% of mediator release from neurons. Preferably, the chimeric Clostridial neurotoxin inhibits 100% of mediator release from neurons.

[0029] The pain mediator may be a neurotransmitter.

[0030] The inhibition of pain mediator release from neurons may be partial or complete, preferably complete. For example, the chimeric Clostridial neurotoxin may inhibit at least 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% of pain mediator release from neurons. Preferably, the chimeric Clostridial neurotoxin inhibits 100% of pain mediator release from neurons.

[0031] The inhibition is preferably inhibition of SNARE-associated (eg, SNAP25-associated) release.

[0032] The chimeric clostridial neurotoxins of the present invention preferably inhibit mediator release from neurons to a greater extent than BoNT / A (preferably, native BoNT / A shown as SEQ ID NO: 6 [e.g., the two-chain form of SEQ ID NO: 6]) inhibits mediator release from neurons. At a given dose (e.g., 1 nM), the chimeric clostridial neurotoxins of the present invention can inhibit mediator release from neurons by at least 10% or 20% (preferably, at least 30%) more than the same dose (e.g., 1 nM) of BoNT / A. At a given dose (e.g., 1 nM), the chimeric clostridial neurotoxins of the present invention can inhibit mediator release from neurons by 10 to 90% or 20 to 90% (preferably, 30 to 85%) more than the same dose (e.g., 1 nM) of BoNT / A. Thus, compared to BoNT / A, a much smaller dose of the chimeric clostridial neurotoxin can inhibit the release of the same amount of mediator from neurons. For example, the dose of the chimeric Clostridial neurotoxin can be reduced by at least 100-fold, 200-fold, or 500-fold, preferably 1000-fold, compared to the dose of BoNT / A required to inhibit mediator release from the same amount of neurons. For example, the dose of the chimeric Clostridial neurotoxin can be reduced by at least 500-2000-fold or 750-1750-fold, preferably 1000-1500-fold, compared to the dose of BoNT / A required to inhibit mediator release from the same amount of neurons.

[0033] The chimeric clostridial neurotoxins of the present invention preferably inhibit the release of pain mediators from neurons to a greater extent than BoNT / A (preferably, native BoNT / A shown as SEQ ID NO: 6 [e.g., the two-chain form of SEQ ID NO: 6]) inhibits the release of pain mediators from neurons. At a given dose (e.g., 1 nM), the chimeric clostridial neurotoxins of the present invention can inhibit the release of pain mediators from neurons by at least 10% or 20% (preferably, at least 30%) more than the same dose (e.g., 1 nM) of BoNT / A. At a given dose (e.g., 1 nM), the chimeric clostridial neurotoxins of the present invention can inhibit the release of pain mediators from neurons by 10 to 90% or 20 to 90% (preferably, 30 to 85%) more than the same dose (e.g., 1 nM) of BoNT / A. Thus, compared to BoNT / A, a much smaller dose of the chimeric clostridial neurotoxin can inhibit the release of the same amount of pain mediators from neurons. For example, the dose of the chimeric Clostridial neurotoxin can be reduced by at least 100-fold, 200-fold, or 500-fold, preferably 1000-fold, compared to the dose of BoNT / A required to inhibit the release of pain mediators from the same number of neurons. The dose of the chimeric Clostridial neurotoxin can be reduced by at least 500-2000-fold or 750-1750-fold, preferably 1000-1500-fold, compared to the dose of BoNT / A required to inhibit the release of pain mediators from the same number of neurons.

[0034] The chimeric clostridial neurotoxins can inhibit the release of multiple mediators from neurons.

[0035] Chimeric clostridial neurotoxins can inhibit the release of multiple pain mediators from neurons.

[0036] The chimeric clostridial neurotoxins of the present invention preferably have analgesic properties, in other words, the chimeric clostridial neurotoxins of the present invention are preferably analgesic chimeric clostridial neurotoxins.

[0037] Preferably, the chimeric Clostridial neurotoxins of the present invention do not promote neuronal growth or neuronal repair to treat pain, in other words, the chimeric Clostridial neurotoxins preferably do not treat pain by either promoting neuronal growth, promoting neuronal repair, or promoting neuronal growth and neuronal repair.

[0038] Preferably, the chimeric Clostridial neurotoxins of the present invention do not promote neuronal growth or promote neuronal repair to treat the disorders described herein, in other words, preferably, the chimeric Clostridial neurotoxins do not treat the disorders described herein by either promoting neuronal growth, promoting neuronal repair, or promoting neuronal growth and neuronal repair.

[0039] The term "promoting neuron growth and / or neuronal repair" encompasses increasing the rate of neuron growth and / or neuronal repair. The term "neuron growth and / or neuronal repair" encompasses rebuilding damaged neuronal circuits, thereby restoring activity and / or neuronal communication in a network or population of neurons. Thus, the term "neuronal repair," as used herein, encompasses repair of specific neurons as well as repair of neuronal circuits. Neuronal plasticity is also encompassed by this term. The term "neuronal plasticity," as used herein, encompasses axonal sprouting, dendritic sprouting, neurogenesis (e.g., the generation of new neurons), maturation, differentiation, and / or synaptic plasticity (e.g., changes in synaptic strength, activity, anatomical morphology, and / or connectivity). The term "promoting neuron growth and / or neuronal repair" also encompasses promoting the establishment of functional synapses (e.g., at or near the site of injury). The term "neuron growth," as used herein, encompasses the growth of any part of a neuron, including axonal and / or dendrite growth. The term encompasses an increase in neurite length, neurite number (e.g., number of neurites per cell), and / or length and / or number of processes from the neuronal cell body or cell membrane, e.g., neuronal axonal growth and / or axonal sprouting, in neurons of interest, etc. The axonal growth can facilitate connections and / or chemical communication between neurons.

[0040] Preferably, the chimeric Clostridial neurotoxins of the present invention do not promote a neuroimmune response for treating pain. Preferably, the chimeric Clostridial neurotoxins of the present invention do not promote a neuroimmune response for treating the disorders described herein. A neuroimmune response in this context includes a microglial response. Thus, in one embodiment, the chimeric Clostridial neurotoxins of the present invention do not promote a microglial response for treating pain. Thus, in one embodiment, the chimeric Clostridial neurotoxins of the present invention do not promote a microglial response for treating the disorders described herein.

[0041] In preferred embodiments, the pain is not pain associated with or caused by a brain disorder. In preferred embodiments, the disorders described herein are not disorders associated with or caused by a brain disorder. The term "brain disorder" as used in this context is interchangeable with "brain disease." "Brain disorder," as used in this context, encompasses disorders originating within or outside the brain, and includes disorders associated with physical injuries that cause brain tissue damage. Examples of brain disorders encompassed in this context include any one or more of traumatic brain injury, cancer (e.g., brain tumor), infectious disease (e.g., encephalitis, meningitis, brain abscess, and encephalitis), stroke, neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson's disease, Parkinson's disease-related disorders, motor neuron diseases (e.g., amyotrophic lateral sclerosis), prion diseases, Huntington's disease, spinocerebellar ataxia, ataxia, Hallervorden-Spatz disease, and frontotemporal lobar degeneration), cerebral aneurysm, multiple sclerosis, anoxic injury, toxic injury, and metabolic injury. Brain disorders can result from traumatic brain injury, cancer, infectious diseases (e.g., encephalitis, meningitis, brain abscess, and encephalitis), stroke, neurodegenerative disorders (e.g., Alzheimer's disease, Parkinson's disease, Parkinson's disease-related disorders, motor neuron diseases (e.g., amyotrophic lateral sclerosis), prion diseases, Huntington's disease, spinocerebellar ataxia, ataxia, Hallervorden-Spatz disease, and frontotemporal lobar degeneration), cerebral aneurysm, multiple sclerosis, anoxic injury, toxic injury, and / or metabolic injury.

[0042] The chimeric clostridial neurotoxin preferably binds to neurons containing Aδ fibers or C fibers. The binding is due to the BoNT / BH activity of the chimeric clostridial neurotoxin. C domain (e.g., its H CC After binding to a neuron, the chimeric Clostridial neurotoxin is internalized via the endosome, and the BoNT / A light chain can be translocated from the endosome into the cytosol of the neuron by the BoNT / A translocation domain. Once in the cytosol, the light chain can cleave SNARE proteins (e.g., SNAP25), thereby inhibiting release / secretion from the neuron (including release / secretion of pain mediators from the neuron).

[0043] Neurons containing A-delta fibers or C fibers are described in Pichon & Chesler (2014), Frontiers in Neuroanatomy (https: / / doi.org / 10.3389 / fnana.2014.00021) and Yam et al. (2018), Int J Mol Sci, 19, 8, 2164. The term "fiber" (e.g., in the context of A-delta fibers or C fibers) preferably refers to the axon of a neuron. Generally, multiple fibers (e.g., multiple A-delta fibers or multiple C fibers, respectively) together can define a larger nerve / neuron structure in a subject, e.g., as a fiber bundle. For example, an A-delta fiber bundle or a C-fiber bundle. In some embodiments, the multiple fibers can also include fibers other than A-delta fibers or C fibers. For example, a nerve can include multiple neurons, including neurons containing A-delta fibers and / or neurons containing C fibers.

[0044] A chimeric clostridial neurotoxin can bind to neurons containing Aδ fibers. Aδ fibers (or neurons containing Aδ fibers) can be characterized as peptidergic, fast-conducting, lightly myelinated, involved in sharp / fast pain, nociception, and / or temperature sensation. Preferably, Aδ fibers (or neurons containing Aδ fibers) can have a conduction velocity of 5 to 75 m / s (e.g., 5 to 35 m / s) and / or a diameter of about 1 to 5 μm (e.g., 2 to 5 μm). Aδ fiber-containing neurons to which the chimeric clostridial neurotoxin of the present invention binds are neurons that can release pain mediators. In particular, such neurons can release CGRP and thus may play a role in CGRP-related pain. By binding to Aδ fiber-containing neurons, the chimeric clostridial neurotoxin inhibits the release of pain mediators from the neurons by cleaving their SNARE proteins (e.g., SNAP25), thereby inhibiting the release / secretion of pain mediators from the neurons.

[0045] A chimeric clostridial neurotoxin can bind to neurons containing C fibers. C fibers (or neurons containing C fibers) can be characterized as peptidergic, low (e.g., slow) conduction, unmyelinated, involved in dull / slow pain, neuropathic pain, temperature sensation, and / or itch sensation. C fiber-containing neurons can be polymodal. Preferably, C fibers (or neurons containing C fibers) can have a conduction velocity of 0.5 to 2 m / s and / or a diameter of about 0.2 to 1.5 μm (e.g., 0.2 to 0.5 μm). C fiber-containing neurons to which the chimeric clostridial neurotoxin of the present invention binds are neurons that can release pain mediators. In particular, such neurons can release CGRP and thus may play a role in CGRP-related pain. By binding to neurons containing C-fibers, the chimeric clostridial neurotoxin can inhibit the release of pain mediators from said neurons by cleaving their SNARE proteins (e.g., SNAP25), thereby inhibiting the release / secretion of pain mediators from said neurons.

[0046] Preferably, the neurons to which the chimeric Clostridial neurotoxin binds are neurons that contain C-fibers.

[0047] Expression of tropomyosin receptor kinase A (TrkA) may be a marker for distinguishing neurons containing Aδ or C nerve fibers (e.g., from neurons containing Aβ fibers). In other words, neurons containing Aδ or C nerve fibers of the present invention may be neurons that express TrkA.

[0048] In use, the chimeric Clostridial neurotoxin may bind to multiple neurons, including at least an A-delta fiber-containing neuron and a C-fiber-containing neuron. The multiple neurons may be part of a larger nerve / neuronal structure in the subject (e.g., comprising a fiber bundle).

[0049] Neurons containing Aδ or C nerve fibers may be neurons of the central nervous system (e.g., hypothalamus, thalamus, periaqueductal gray matter, superior colliculus, inferior colliculus, amygdala, trigeminocervical complex, and / or cerebellum) or neurons of the peripheral nervous system. The chimeric clostridial neurotoxins can inhibit the release of mediators from neurons of the central nervous system when treating certain conditions, such as headache, preferably migraine pain. The chimeric clostridial neurotoxins can inhibit the release of pain mediators from neurons of the central nervous system when treating certain pain conditions, such as headache, preferably migraine pain.

[0050] Neurons comprising Aδ or C nerve fibers according to the present invention are preferably sensory neurons. The sensory neurons may be primary sensory neurons, such as primary afferent neurons. For example, the neurons to which the chimeric Clostridial neurotoxin binds may be sensory neurons of the dorsal root ganglion and / or the trigeminal ganglion. Additionally or alternatively, the neurons may be intrinsic enteric neurons.

[0051] A chimeric clostridial neurotoxin of the invention can bind to neurons containing A-delta fibers or C-fibers with greater affinity than the affinity with which BoNT / A (preferably, native BoNT / A shown as SEQ ID NO: 6 [e.g., the two-chain form of SEQ ID NO: 6]) binds to neurons. In particular, a chimeric clostridial neurotoxin of the invention can bind to neurons containing A-delta fibers or C-fibers with an affinity that is at least 2-fold, 5-fold, 10-fold, 50-fold, 100-fold, 1,000-fold, or 10,000-fold greater than the affinity with which BoNT / A binds to neurons.

[0052] The chimeric Clostridial neurotoxins of the present invention can bind to neurons containing Aδ fibers or C fibers with greater affinity than the affinity with which the chimeric Clostridial neurotoxin binds to neurons (preferably sensory neurons) that do not contain Aδ fibers or C fibers (e.g., neurons containing Aβ fibers). For example, the chimeric Clostridial neurotoxins of the present invention can bind to neurons containing Aδ fibers or C fibers with an affinity that is at least 2-fold, 5-fold, 10-fold, 50-fold, 100-fold, 1,000-fold, or 10,000-fold greater than the affinity with which the chimeric Clostridial neurotoxin binds to neurons (preferably sensory neurons) that do not contain Aδ fibers or C fibers (e.g., sensory neurons that contain Aβ fibers).

[0053] Aβ fibers (or neurons comprising Aβ fibers) can be characterized as myelinated, fast-conducting, involved in touch, and / or generally responsive to other non-noxious stimuli. Preferably, Aβ fibers (or neurons comprising Aβ fibers) can have a conduction velocity of 80-120 m / s and / or a diameter of about 6-20 μm. Expression of neurofilament 200 (NF200) can be a marker for distinguishing neurons comprising Aβ nerve fibers (e.g., from neurons comprising Aδ fibers or C fibers). In other words, neurons comprising Aβ fibers can be neurons that express NF200.

[0054] In other embodiments, the chimeric clostridial neurotoxin may exert its effect at a site distal to the site of administration (e.g., injection). For example, after administration of the chimeric clostridial neurotoxin, SNARE protein cleavage (e.g., SNAP25 cleavage) may occur at a site distal to the site of administration (e.g., injection). Preferably, such an effect occurs via neuronal transport of the chimeric clostridial neurotoxin from the site of administration to a site distal to the site of administration. When treating pain (preferably headache, most preferably migraine pain) or migraine, the chimeric clostridial neurotoxin preferably exerts its effect at a site distal to the site of administration (e.g., injection). In one embodiment, this effect may be in addition to a peripheral effect. Thus, preferably, when treating pain (preferably headache, most preferably migraine pain) or migraine, the chimeric clostridial neurotoxin may be transported via neuronal transport.

[0055] Neuronal transport may be retrograde or anterograde, preferably retrograde. Transport may be axonal transport.

[0056] "Retrograde transport" can be a form of axonal transport (also known as intra-axonal transport or axonal flow). Axonal transport is the cellular process responsible for the movement of mitochondria, lipids, synaptic vesicles, proteins, and other organelles to and from the neuronal cell body through the cytoplasm of the axon, usually called the axoplasm. Axons are several meters long, and neurons use axonal transport because they cannot transport nuclear and organelle products to the axon terminals by diffusion. Axonal transport also is responsible for the movement of molecules from the axon back to the cell body that are destined for degradation in the cell body by lysosomes.

[0057] "Retrograde transport" can refer to movement toward the cell body of a neuron, and "anterograde transport" can refer to movement toward the synapse of a neuron.

[0058] In one embodiment, neuronal (e.g., retrograde) transport to a neuron of the central nervous system may refer to transport (e.g., axonal transport) of a chimeric Clostridial neurotoxin toward a neuronal cell body, the neuronal cell body being located near the central nervous system.

[0059] Neuronal (e.g., retrograde) transport will now be described in more detail. In one embodiment, the chimeric clostridial neurotoxin can bind to a first neuron (such as a primary sensory afferent nerve) at the administration site. The chimeric clostridial neurotoxin can be internalized by the first neuron, transported within the first neuron, and then released from the first neuron. Preferably, the clostridial neurotoxin binds to the first neuron at an intramuscular or intradermal administration site (e.g., intramuscular or intradermal injection). Such neurons can be peripheral neurons, preferably neurons containing A-delta fibers or C fibers. Once released, the chimeric clostridial neurotoxin can bind to a second neuron, be internalized, and cleave a SNARE protein (e.g., SNAP25) within the second neuron. Alternatively, the chimeric clostridial neurotoxin can bind to a second neuron, be internalized by the second neuron, be transported within the second neuron, and then be released from the second neuron. This process may be repeated until the chimeric Clostridial neurotoxin binds to a neuron (e.g., a third neuron), is internalized, and cleaves a SNARE protein (e.g., SNAP25) within the neuron. The second neuron may be a second sensory afferent. Preferably, the second neuron is a neuron of the central nervous system, such as a neuron located in the brain, brainstem, or spinal cord. The second neuron may be a neuron located in the trigeminal ganglion (e.g., SNARE cleavage may occur in its axon).

[0060] In some embodiments, upon intramuscular administration, the chimeric Clostridial neurotoxin may be transported neuronally (e.g., retrogradely) through a motor neuron, released from the motor neuron, and enter a second neuron, preferably a neuron of the central nervous system, which may be a sensory neuron.

[0061] In one embodiment, upon intramuscular administration, the chimeric Clostridial neurotoxin can diffuse to and bind to (eg, terminate in) sensory neurons present in the periosteum or skin.

[0062] Without wishing to be bound by theory, it is believed that the chimeric clostridial neurotoxins of the present invention can block secretion from one or more neurons in the central nervous system through the aforementioned neuronal (e.g., retrograde) transport mechanism. Thus, the chimeric clostridial neurotoxin can travel to neurons in the central nervous system by neuronal (e.g., retrograde) transport and cleave SNARE proteins (e.g., SNAP25) in the neurons.

[0063] In preferred embodiments, the chimeric clostridial neurotoxin can treat pain or a disorder described herein by inhibiting secretion (e.g., inhibiting the release of a mediator (e.g., a pain mediator)) from one or more neurons in the central nervous system. This is particularly relevant in the treatment of pain or migraine, preferably in the treatment of migraine pain. The chimeric clostridial neurotoxin can travel to neurons in the central nervous system by neuronal (e.g., retrograde) transport and cleave SNARE proteins (e.g., SNAP25) in the neurons. Thus, the chimeric clostridial neurotoxin can treat pain (e.g., headache or migraine pain) or migraine by inhibiting secretion from neurons in the central nervous system, preferably by inhibiting the secretion of a mediator, more preferably a pain mediator, from neurons in the central nervous system.

[0064] Neurons of the central nervous system may be neurons of the brainstem, spinal cord, and / or brain. For example, neurons of the central nervous system may be neurons of the trigeminal nucleus (e.g., spinal trigeminal nucleus such as the spinal trigeminal sensory nucleus), spinal cord (preferably neurons of the dorsal horn of the spinal cord), hypothalamus, thalamus, periaqueductal gray matter, superior colliculus, inferior colliculus, amygdala, cervical trigeminal complex, cortex, and / or cerebellum. Neurons of the trigeminal nucleus may be neurons of the trigeminal nucleus caudalis (e.g., pars caudalis).

[0065] In a preferred embodiment, the chimeric clostridial neurotoxin cleaves a SNARE protein (e.g., SNAP25) in neurons of the brainstem, more preferably in neurons of the trigeminal nucleus (even more preferably, the spinal (sensory) nucleus of the trigeminal nerve). The chimeric clostridial neurotoxin can inhibit secretion (e.g., of a mediator, preferably a pain mediator) from the neuron. Cleavage of the SNARE protein can occur via neuronal (e.g., retrograde) transport of the chimeric clostridial neurotoxin from the administration site. Such neurons can be targeted by administering the chimeric clostridial neurotoxin to muscle, periosteum, and / or skin innervated by sensory trigeminal neurons (e.g., the muscle, periosteum, and / or facial skin and / or scalp of a subject). Alternatively, the neuron may comprise Aδ or C nerve fibers, to which the chimeric clostridial neurotoxin can bind and subsequently cleave the SNARE protein (e.g., after transport / diffusion through the cytoplasm of the neuron). The cleavage can occur at the terminal of a neuron in the spinal sensory trigeminal nucleus. Most preferably, inhibiting the cleavage and secretion of said SNAREs treats migraine or migraine pain.

[0066] In one embodiment, the chimeric clostridial neurotoxin cleaves a SNARE protein (e.g., SNAP25) in neurons of the trigeminal motor nucleus. The chimeric clostridial neurotoxin can inhibit secretion from the neurons. Cleavage of the SNARE protein can occur via neuronal (e.g., retrograde) transport of the chimeric clostridial neurotoxin from the administration site.

[0067] In another preferred embodiment, the chimeric clostridial neurotoxin cleaves a SNARE protein (e.g., SNAP25) in neurons of the spinal cord, such as the cervical spinal cord. More preferably, the neurons are neurons in the dorsal horn of the spinal cord (e.g., associated with sensory neurons). The chimeric clostridial neurotoxin can inhibit secretion (e.g., of mediators, preferably pain mediators) from the neurons. Cleavage of the SNARE protein can occur via neuronal (e.g., retrograde) transport of the chimeric clostridial neurotoxin from the administration site. Such neurons can be targeted by administering the chimeric clostridial neurotoxin to muscle, periosteum, and / or skin innervated by spinal sensory neurons (e.g., muscle, periosteum, and / or skin of the back of the head and / or neck of a subject). Alternatively, the neurons may include Aδ or C nerve fibers, to which the chimeric clostridial neurotoxin can bind and subsequently cleave the SNARE protein (e.g., after transport / diffusion through the cytoplasm of the neuron). The cleavage can also occur at the neuron terminal in the spinal cord. Most preferably, inhibiting the cleavage and secretion of said SNAREs treats migraine or migraine pain.

[0068] In one embodiment, the chimeric clostridial neurotoxin cleaves a SNARE protein (e.g., SNAP25) in ventral horn neurons of the spinal cord (e.g., associated with motor neurons). The chimeric clostridial neurotoxin can inhibit secretion (e.g., of mediators, preferably pain mediators) from the neurons. Cleavage of the SNARE protein can occur via neuronal (e.g., retrograde) transport of the chimeric clostridial neurotoxin from the site of administration.

[0069] In another preferred embodiment, the chimeric clostridial neurotoxin cleaves a SNARE protein (e.g., SNAP25) in trigeminal ganglion neurons, e.g., in their axons. The chimeric clostridial neurotoxin can inhibit secretion (e.g., of mediators, preferably pain mediators) from the neurons. Cleavage of the SNARE protein can occur via neuronal (e.g., retrograde) transport of the chimeric clostridial neurotoxin from the administration site. Alternatively, the neuron may contain Aδ or C nerve fibers, to which the chimeric clostridial neurotoxin can bind and subsequently cleave the SNARE protein (e.g., after transport / diffusion through the neuronal cytoplasm). Most preferably, inhibition of SNARE cleavage and secretion treats migraine or migraine pain.

[0070] Neuronal (e.g., retrograde) transport of clostridial neurotoxins has been described (see Bomba-Warczak et al. (2016), Cell Rep., 16(7), 1974-1987), and without wishing to be bound by theory, it is believed to occur through binding of the clostridial neurotoxin to a non-canonical receptor (e.g., in the present case, via binding to a receptor other than SYTI or SYTII), incorporation into non-acidified organelles, neuronal (e.g., retrograde) transport (e.g., from the periphery of the body toward the central nervous system), and release from the neuron into the extracellular space. In such cases, the clostridial neurotoxin is described as remaining intact (i.e., the two chains comprising the light and heavy chains linked to each other by a disulfide bond remain intact), allowing it to bind to a second neuron via the canonical intoxication pathway (e.g., via SYTI or SYTII in the context of the chimeric clostridial neurotoxin of the present invention).

[0071] A portion of the chimeric clostridial neurotoxin administered to a subject binds to neurons containing Aδ nerve fibers or C nerve fibers and inhibits the release of mediators (e.g., pain mediators) from the neurons, and the portion of the chimeric clostridial neurotoxin can exert its effect at a site distal to the administration site. The portion capable of exerting its effect at a site distal to the administration site can inhibit secretion from neurons in the central nervous system, preferably the secretion of mediators (e.g., neurotransmitters), more preferably the secretion of pain mediators from neurons in the central nervous system. The chimeric clostridial neurotoxin can travel to neurons in the central nervous system by neuronal (e.g., retrograde) transport and cleave SNARE proteins (e.g., SNAP25) in the neurons.

[0072] In some embodiments, neuronal (eg, retrograde) transport of the chimeric clostridial neurotoxin can include transsynaptic movement (eg, transcytosis) of the chimeric clostridial neurotoxin from one neuron to another neuron.

[0073] The inhibition of neuronal secretion may be partial or complete, preferably complete. For example, the chimeric Clostridial neurotoxin may inhibit neuronal secretion by at least 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99%. Preferably, the chimeric Clostridial neurotoxin inhibits neuronal secretion by 100%. In this context, secretion is preferably SNARE-associated (e.g., SNAP25-associated) secretion.

[0074] In preferred embodiments, the chimeric Clostridial neurotoxins of the present invention may treat migraine or a disorder described herein (preferably, pain) by inhibiting the release of a mediator (e.g., a pain mediator) from neurons comprising Aδ nerve fibers or C nerve fibers (wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively), and by inhibiting secretion (e.g., of a mediator, preferably a pain mediator) from neurons in the central nervous system.

[0075] Bacteria of the genus Clostridium produce highly potent and specific protein toxins that can harm neurons and other cells to which they are delivered. Examples of such clostridial toxins include those produced by Clostridium baratii and Clostridium butyricum, as well as those produced by Clostridium tetani (TeNT) and Clostridium botulinum (BoNT) serotypes A to G and X (see WO 2018 / 009903 A2). Both tetanus and botulinum toxins act by inhibiting the function of affected neurons, particularly by inhibiting the release of neurotransmitters. Botulinum toxins generally act at the neuromuscular junction and inhibit cholinergic transmission in the peripheral nervous system, while tetanus toxins act in the central nervous system.

[0076] In nature, clostridial neurotoxins are synthesized as single-chain polypeptides that are post-translationally modified by a proteolytic cleavage event to form two polypeptide chains linked together by a disulfide bond. Cleavage occurs at a specific cleavage site, often referred to as the activation site (e.g., activation loop), located between cysteine ​​residues that provide the interchain disulfide bond. This two-chain form is the active form of the toxin. The two chains are termed heavy chains (H chains) with a molecular weight of approximately 100 kDa and light chains (L chains) with a molecular weight of approximately 50 kDa. The H chains are linked to an N-terminal translocation component (H N domain) and C-terminal targeting component (H C The cleavage site is located between the L chain and the translocation domain component. C After binding of the domain to its target neuron and internalization of the bound toxin into the cell via endosomes, H N The domain translocates the light chain across the endosomal membrane into the cytosol, where the light chain provides the protease function (also known as a non-cytotoxic protease).

[0077] The non-cytotoxic protease acts by proteolytically cleaving intracellular transport proteins known as SNARE proteins (e.g., SNAP25, VAMP, or syntaxin, preferably SNAP25). S oluble N science fiction A Attachment Re The name comes from receptor, and NSF is N -ethylmaleimide- S sensitive F The term "actor" refers to a specific protein that is a cytotoxic agent. SNARE proteins are essential for intracellular vesicle fusion and therefore for the secretion of molecules from cells via vesicle transport. The protease function is a zinc-dependent endopeptidase activity that exhibits high substrate specificity for SNARE proteins. Thus, once delivered to desired target cells, the non-cytotoxic protease can inhibit cellular secretion from the target cells. The L-chain proteases of clostridial neurotoxins are non-cytotoxic proteases that cleave SNARE proteins.

[0078] Due to the ubiquitous nature of SNARE proteins, clostridial neurotoxins, such as botulinum toxin, have been used successfully in a wide range of therapeutic modalities.

[0079] For further details on the genetic basis of toxin production in Clostridium botulinum and Clostridium tetani, see Henderson et al (1997) The Clostridia: Molecular Biology and Pathogenesis, Academic Press.

[0080] Clostridial neurotoxin domains are described in more detail below.

[0081] Examples of reference sequences for the light chain include the following: Botulinum type A neurotoxin: amino acid residues 1 to 448 Botulinum type B neurotoxin: amino acid residues 1 to 440

[0082] The reference sequences provided above should be considered as a guideline and slight variations may occur depending on the subserotype. For example, US 2007 / 0166332 (hereby incorporated by reference in its entirety) lists slightly different Clostridial sequences. Botulinum type A neurotoxin: amino acid residues M1 to K448 Botulinum type B neurotoxin: amino acid residues M1 to K441

[0083] The translocation domain is a fragment of the heavy chain of a clostridial neurotoxin that is approximately equivalent to the amino-terminal half of the heavy chain, or the domain corresponding to that fragment in an intact heavy chain.

[0084] Examples of reference translocation domains include: Botulinum type A neurotoxin: amino acid residues (449 to 871) Botulinum type B neurotoxin: amino acid residues (441 to 858)

[0085] The reference sequences given above should be considered as a guideline and slight variations may occur depending on the subserotype. For example, US 2007 / 0166332 (hereby incorporated by reference) lists slightly different Clostridial sequences. Botulinum type A neurotoxin: amino acid residues (A449 to K871) Botulinum type B neurotoxin: amino acid residues (A442 to S858)

[0086] In the context of the present invention, various BoNT / AHs containing a translocation domain N The H region of the BoNT / A heavy chain may be useful in embodiments of the present invention. N The region is approximately 410 to 430 amino acids long and contains the translocation domain. Studies have shown that the translocation domain translocates to the H of the clostridial neurotoxin heavy chain due to its translocation activity. NIt has been shown that the full length of the region is not necessary. Thus, aspects of this embodiment include, for example, BoNT / AH sequences that include a translocation domain that is at least 350 amino acids, at least 375 amino acids, at least 400 amino acids, or at least 425 amino acids in length. N Other aspects of this embodiment include a BoNT / AH gene that includes a translocation domain that is, for example, at most 350 amino acids, at most 375 amino acids, at most 400 amino acids, or at most 425 amino acids in length. N It may include a region.

[0087] Term H N is a naturally occurring BoNT / AH N Modified BoNT / AHs having portions and non-naturally occurring amino acid sequences and / or synthetic amino acid residues N Preferably, the modified BoNT / AH N The moiety still exhibits said translocation function.

[0088] Clostridial neurotoxin receptor binding domain (H C ) Examples of reference sequences include: BoNT / A-N872 to L1296 BoNT / B-E859 to E1291

[0089] Approximately 50 kDa H of clostridial neurotoxins (BoNT, etc.) C The domains are H, which are usually about 25 kDa each. CC Domain and H CN It has two distinct structural features called domains. The amino acid residues involved in receptor binding are mainly H CC It is thought to be located in the H domain of natural clostridial neurotoxins. CA domain may contain approximately 400 to 440 amino acid residues. This fact is confirmed by the following publications, each of which is incorporated herein by reference in its entirety: Umland TC (1997) Nat. Struct. Biol. 4: 788-792; Herreros J (2000) Biochem. J. 347: 199-204; Halpern J (1993) J. Biol. 104: 359-364; Lacey DB (1998) Nat. Struct. Biol. 5: 898-902; Knapp (1998) Am. Cryst. Assoc. Abstract Papers 25: 90; Swaminathan and Eswaramoorthy (2000) Nat. Struct. Biol. 7: 1751-1759; and Rummel A (2004) Mol. Microbiol. 51(3), 631-643.

[0090] (Reference)H CN Examples of domains include: Botulinum type A neurotoxin: amino acid residues (872 to 1110) Botulinum type B neurotoxin: amino acid residues (859 to 1097)

[0091] The above sequence positions may vary slightly depending on the serotype / subtype. (See H) CN Further examples of domains include: Botulinum type A neurotoxin: amino acid residues (874 to 1110) Botulinum type B neurotoxin: amino acid residues (861 to 1097)

[0092] (Reference)H CC The domains include the following: Botulinum type A neurotoxin: amino acid residues (Y1111 to L1296) Botulinum type B neurotoxin: amino acid residues (Y1098 to E1291)

[0093] WO 2017 / 191315 A1 (incorporated herein by reference) teaches chimeric clostridial neurotoxins and methods for their preparation and manufacture. Accordingly, the botulinum neurotoxin A (BoNT / A) light chain and translocation domain (BoNT / AH) used in the present invention may be used in combination with other clostridial neurotoxins. N domain), and the BoNT / B receptor binding domain (H C The chimeric clostridial neurotoxin comprising the clostridial domain may be one taught in WO 2017 / 191315 A1.

[0094] The term "chimeric clostridial neurotoxin" or "chimeric neurotoxin," as used herein, refers to a chimeric clostridial neurotoxin comprising a clostridial neurotoxin light chain and a translocation domain (H) from a first clostridial neurotoxin serotype. N domain), as well as a receptor-binding domain (H C Specifically, a chimeric Clostridial neurotoxin for use in the present invention refers to a neurotoxin comprising (preferably consisting of) a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H domain). N domain), and the BoNT / B receptor binding domain (H C The chimeric clostridial neurotoxin BoNT / A LH contains N The domain is BoNT / BH C The chimeric clostridial neurotoxins of the present invention may also be referred to as chimeric botulinum neurotoxins. The chimeric clostridial neurotoxins are also referred to herein as "BoNT / AB," "mrBoNT / AB," or "BoNT / AB chimeras."

[0095] L chain and H N The domains (optionally including a complete or partial activation loop, e.g., a complete activation loop if the chimeric clostridial neurotoxin is in single-chain form, or a truncated / partial activation loop if the chimeric clostridial neurotoxin is in two-chain form) are collectively designated LH N It is sometimes called a domain. Therefore, LHN The domain is further H C Does not include the domain.

[0096] The chimeric clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0097] The term "consisting essentially of," as used in this context, means that the chimeric Clostridial neurotoxin does not further comprise one or more amino acid residues that confer additional functionality to the polypeptide, for example, when administered to a subject. In other words, the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N domain), and the BoNT / B receptor binding domain (H C A polypeptide "consisting essentially of" a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H domain) can further comprise a polypeptide "consisting essentially of" a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H domain). N domain), and the BoNT / B receptor binding domain (H C The polypeptide may contain one or more amino acid residues (in addition to those of the corresponding domain), provided that the one or more additional amino acid residues do not confer any additional functionality to the polypeptide, e.g., when administered to a subject. The additional functionality may include enzymatic activity, binding activity, and / or any other physiological activity.

[0098] A chimeric clostridial neurotoxin may contain a non-clostridial neurotoxin sequence in addition to any clostridial neurotoxin sequence, so long as the chimeric clostridial neurotoxin does not interfere with its ability to achieve a therapeutic effect (preferably for the treatment of pain). Preferably, the non-clostridial neurotoxin sequence does not have catalytic activity, such as enzymatic activity. In one embodiment, a chimeric clostridial neurotoxin of the present invention does not contain a non-clostridial catalytically active domain. In one embodiment, a chimeric clostridial neurotoxin does not contain an additional catalytically active domain. In one embodiment, the non-clostridial sequence is not a sequence that binds to a cellular receptor. In other words, in one embodiment, the non-clostridial sequence is not a ligand for a cellular receptor. The cellular receptor may be a proteinaceous cellular receptor, such as an integral membrane protein. Examples of cellular receptors are listed in the IUPHAR Guide to Pharmacology Database, version 2019.4, available at https: / / www.guidetopharmacology.org / download.jsp#db_reports. The non-clostridial neurotoxin sequence may include a tag to aid in purification, such as a histidine tag. In one embodiment, the chimeric clostridial neurotoxin of the invention does not include a label or a site for adding a label, such as a sortase acceptor or donor site.

[0099] Preferably, the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0100] The chimeric clostridial neurotoxin contains a light chain that exhibits non-cytotoxic protease activity and can cleave SNARE proteins in the cytosol of target neurons. As explained above, the two-chain form is the active form of the clostridial neurotoxin. Therefore, the present invention excludes the use of chimeric clostridial neurotoxins containing a light chain that has been catalytically inactivated, for example, by one or more mutations ("catalytically inactive light chains"). Such catalytically inactive light chains (and clostridial neurotoxins containing them) are known in the art. A catalytically inactive light chain may contain one or more mutations that inactivate the catalytic activity. For example, a catalytically inactive light chain may contain a mutation of an active site residue. The mutation may be a substitution or deletion, particularly a substitution with a chemically similar amino acid. Glutamic acid may be substituted with glutamine, histidine may be substituted with tyrosine, arginine may be substituted with glutamine, and / or tyrosine may be substituted with phenylalanine. Alternatively, any residue may be substituted with alanine. A catalytically inactive BoNT / AL chain may contain mutations at H223, E224, H227, E262, R363, and / or Y366, for example, mutations at least E224 and H227. A catalytically inactive BoNT / AL chain may contain a substitution of glutamine at E224 (E224Q) and tyrosine at H227 (H227Y).

[0101] The term "catalytically inactive," as used herein with respect to a clostridial neurotoxin L chain, means that the L chain exhibits substantially no cytotoxic protease activity, e.g., no cytotoxic protease activity. A catalytically inactive clostridial neurotoxin L chain may not cleave proteins of the exocellular fusion apparatus within a target cell. The term "substantially free of cytotoxic protease activity" means that the clostridial neurotoxin L chain has less than 5% of the cytotoxic protease activity of a catalytically active clostridial neurotoxin L chain (preferably the L chain of native BoNT / A shown as SEQ ID NO: 6), e.g., less than 2%, less than 1%, or less than 0.1% of the cytotoxic protease activity of a catalytically active clostridial neurotoxin L chain. Non-cytotoxic protease activity can be measured in vitro by incubating a test clostridial neurotoxin L chain with a SNARE protein and comparing the amount of SNARE protein cleaved by the test clostridial neurotoxin L chain with the amount of SNARE protein cleaved by a catalytically active clostridial neurotoxin L chain (preferably the L chain of native BoNT / A, shown as SEQ ID NO: 6) under the same conditions. The amount of cleaved SNARE protein can be quantified using conventional techniques, such as SDS-polyacrylamide gel electrophoresis and Western blotting. A suitable in vitro assay is described in WO 2019 / 145577 A1, which is incorporated herein by reference.

[0102] Cellular and in vivo assays can also be used to determine whether a clostridial neurotoxin containing an L chain, functional cell binding, and translocation domain has non-cytotoxic protease activity. Assays such as the digit abduction score (DAS) assay, dorsal root ganglion (DRG) assay, spinal cord neuron (SCN) assay, and mouse phrenic nerve hemidiaphragm (PNHD) assay are common in the art. Suitable assays for measuring non-cytotoxic protease activity may be those described in Aoki KR, Toxicon 39: 1815-1820; 2001 or Donald et al. (2018), Pharmacol Res Perspect, e00446, 1-14, which are incorporated herein by reference.

[0103] When administered to a subject, the chimeric clostridial neurotoxin is preferably in an active two-chain form, with the light chain and the heavy chain linked to each other by a disulfide bond. When a clostridial neurotoxin (e.g., a chimeric clostridial neurotoxin) is defined herein by a polypeptide sequence (SEQ ID NO:), the L chain portion of the sequence (SEQ ID NO:) may constitute the first chain of a two-chain clostridial neurotoxin (e.g., a two-chain chimeric clostridial neurotoxin), and the H chain portion of the sequence (SEQ ID NO:) may constitute the first chain of a two-chain clostridial neurotoxin (e.g., a two-chain chimeric clostridial neurotoxin). N and H CThe domains together may constitute the second chain of a di-chain clostridial neurotoxin (e.g., a di-chain chimeric clostridial neurotoxin), in which the first and second chains are linked to each other by a disulfide bond. Those skilled in the art will recognize that a protease may cleave at one or more positions within the activation loop of a clostridial neurotoxin (e.g., a chimeric clostridial neurotoxin), preferably at two positions within the activation loop. When cleavage occurs at two or more positions (preferably two positions) within the activation loop, a subfragment of the C-terminal L chain portion of the sequence may be absent from the di-chain clostridial neurotoxin sequence (e.g., a di-chain chimeric clostridial neurotoxin). For this reason, the sequence of a di-chain clostridial neurotoxin (e.g., a di-chain chimeric clostridial neurotoxin) may differ slightly from the sequence of the corresponding single-chain clostridial neurotoxin (e.g., a single-chain chimeric clostridial neurotoxin). The subfragment may be 1 to 15 amino acids. In particular, in one embodiment, when Lys-C is used to convert a single-chain chimeric clostridial neurotoxin into a two-chain clostridial neurotoxin, the subfragment of the C-terminal L chain portion of the absent sequence may be SEQ ID NO: 15 or 16.

[0104] LH N The C-terminal amino acid residue of the domain is the LH of BoNT / A. N Domain and H C Separating Domains 3 10 may correspond to the first amino acid residue of the helix, H C The N-terminal amino acid residue of the domain is the LH of BoNT / B. N Domain and H C Separating Domains 3 10 It may correspond to the second amino acid residue of the helix.

[0105] An example of a BoNT / A polypeptide sequence is set forth in SEQ ID NO:6.

[0106] An example of a BoNT / B polypeptide sequence is set forth in SEQ ID NO: 7 (UniProt Accession No. B1INP5).

[0107] As used herein, "BoNT / A LH N Domain and H C Separating Domains 3 10 When referring to the "first amino acid residue of the helix," N Domain and H C Separating Domains 3 10 It refers to the N-terminal residue of the helix.

[0108] As used herein, "BoNT / B LH N Domain and H C Separating Domains 3 10 When referring to the "second amino acid residue of the helix," N Domain and H C Separating Domains 3 10 It refers to the amino acid residue following the N-terminal residue of the helix.

[0109] "3 10 A "helix" is a type of secondary structure found in proteins and polypeptides, along with α-helices, β-sheets, and reverse turns. 3 10 The amino acids in the helix are arranged in a right-handed helix, with each full turn completed by 10 atoms separating three residues and the intramolecular hydrogen bonds between them. Each amino acid has 10 atoms in the ring formed by the creation of hydrogen bonds, corresponding to a 120° turn in the helix (i.e., the helix has three residues per turn) and a translational movement of 2.0 Å (=0.2 nm) along the helix axis. Most importantly, the NH group of an amino acid forms a hydrogen bond with the C=O group of the amino acid three residues before it, and this repeated i+3→i hydrogen bond is repeated 3 times. 10 Define a helix. 3 10 A helix is ​​a standard concept in structural biology with which those skilled in the art are familiar.

[0110] These three 10 The helix corresponds to the four residues that form the actual helix and two cap (or transition) residues, one at each end of these four residues. N Domain and HC Separating Domains 3 10 A "helix," as used herein, consists of those six residues.

[0111] By performing structural analysis and sequence alignment, LH N Domain and H C Separating Domains 3 10 The three helices were identified. 10 The helix is ​​located at its N-terminus (i.e., LH N by an α-helix at the C-terminal part of the domain) and at its C-terminus (i.e., H C The N-terminal part of the domain is surrounded by β-strands. 10 The first (N-terminal) residue of a helix (the cap or transition residue) also corresponds to the C-terminal residue of this α-helix.

[0112] LH N Domain and H C Separating Domains 3 10 The helices can be determined, for example, from publicly available crystal structures of botulinum neurotoxins, such as 3BTA (http: / / www.rcsb.org / pdb / explore / explore.do?structureId=3BTA) and 1EPW (http: / / www.rcsb.org / pdb / explore / explore.do?structureId=1EPW) for botulinum neurotoxins A1 and B1, respectively.

[0113] LH in other neurotoxins N Domain and H C Separating Domains 3 10Publicly available computer modeling and alignment tools can also be used to determine helix positions, such as the homology modeling servers LOOPP (Learning, Observing and Outputting Protein Patterns, http: / / loopp.org), PHYRE (Protein Homology / analogY Recognition Engine, http: / / www.sbg.bio.ic.ac.uk / phyre2 / ), and Rosetta (https: / / www.rosettacommons.org / ), the protein superposition server SuperPose (http: / / wishart.biology.ualberta.ca / superpose / ), the alignment program Clustal Omega (http: / / www.clustal.org / omega / ), and many other tools / services listed in Internet Resources for Molecular and Cell Biologists (http: / / molbiol-tools.ca / ). In particular, "H N / H CN The perijunctional region can be highly conserved structurally, making it an ideal region for overlapping different serotypes.

[0114] For example, this 3 in other neurotoxins 10 To determine the sequence of the helix, the following methodology may be used. 1. Based on the BoNT / A1 crystal structure (3BTA.pdb), the structural homology modeling tool LOOP (http: / / loopp.org) may be used to obtain predicted structures of other BoNT serotypes. 2. Copy the resulting structure (pdb) file to H CN The N-terminal part of the domain and the approximately 80 residues preceding it (H N domain), thereby resulting in a highly structurally conserved "H N / H CN " area can be edited to preserve it. 3. The protein superposition server SuperPose (http: / / wishart.biology.ualberta.ca / superpose / ) may be used to superimpose each serotype onto the 3BTA.pdb structure. 4. Inspect the overlapping pdb files to identify the H of BoNT / A1. C 3 at the beginning of the domain 10 The helices may be located and then the corresponding residues in other serotypes identified. 5. Clustal Omega may be used to align other BoNT serotype sequences to check for accurate corresponding residues.

[0115] LH determined by this method N , H C , and 3 10 Examples of helical domains are shown in the table below. [Table A]

[0116] By using structural analysis and sequence alignment, LH N Domain and H C Separating Domains 3 10 The β-strand following the helix is ​​a conserved structure in all botulinum and tetanus neurotoxins, and LH N Domain and H C Separating Domains 3 10 It was found to start at the eighth residue (eg, residue 879 of BoNT / A1) when starting from the first residue of the helix.

[0117] BoNT / AB chimera is a chimera of BoNT / B C LH of BoNT / A covalently bound to the domain N domain, where LH N The C-terminal amino acid residue of the domain is H of BoNT / A. CIt corresponds to the eighth amino acid residue at the N-terminus of the β-strand located at the beginning (N-terminus) of the domain, where H C The N-terminal amino acid residue of the domain is the H C It corresponds to the seventh amino acid residue at the N-terminus of the β-strand located at the beginning (N-terminus) of the domain.

[0118] BoNT / AB chimera is a chimera of BoNT / B C LH of BoNT / A covalently bound to the domain N domain, where LH N The C-terminal amino acid residue of the domain is the LH of BoNT / A. N corresponds to the C-terminal amino acid residue of the α-helix located at the end (C-terminus) of the domain, where H C The N-terminal amino acid residue of the domain is the LH of BoNT / B. N It corresponds to the amino acid residue immediately C-terminal to the C-terminal amino acid residue of the α-helix located at the end (C-terminus) of the domain.

[0119] The rationale for the design process of the BoNT / AB chimera was to ensure that the secondary structure was intact, thereby minimizing any changes in the tertiary structure and function of each domain. Without wishing to be bound by theory, it is believed that the 3 10 It is hypothesized that not disrupting the four central amino acid residues of the helix will ensure an optimal conformation of the chimeric neurotoxin, thereby allowing it to maximize its functionality. Indeed, surprisingly, the 3 amino acid residues of BoNT / A 10 The first amino acid residue of the helix and the first three amino acids of BoNT / B 10 Retaining only the second amino acid residue of the helix not only enables the production of a soluble and functional BoNT / AB chimera, but also provides improved properties over other BoNT / AB chimeras, particularly enhanced potency, improved safety margin and / or longer duration of action (and improved safety margin and / or longer duration of action compared to native BoNT / A [e.g., SEQ ID NO: 6]).

[0120] Undesirable effects of neurotoxins (caused by neurotoxin diffusion away from the injection site) can be experimentally assessed by measuring the rate of weight loss in relevant animal models (e.g., mice, where weight loss is detected within 7 days of injection). Conversely, the desired on-target effects of neurotoxins can be experimentally assessed by the digital abduction score (DAS) assay, which measures muscle paralysis. The DAS assay can be performed by injecting 20 μL of neurotoxin formulated in gelatin phosphate buffer into the mouse gastrocnemius / soleus muscle complex, followed by assessing the digital abduction score using the Aoki method (Aoki KR, Toxicon 39: 1815-1820; 2001). In the DAS assay, mice are briefly suspended by their tails to elicit a characteristic startle response in which the mice extend their hind limbs and abduct their hind digits. After neurotoxin injection, the degree of digital abduction is scored on a 5-point scale (0 = normal to 4 = maximal reduction in digital abduction and leg extension).

[0121] The safety factor of a neurotoxin can then be expressed as the ratio of the amount of neurotoxin required to reduce a mouse's body weight by 10% (measured at peak effect within the first 7 days after dosing the mouse) to the amount of neurotoxin required for a DAS score of 2. Thus, a high safety factor score is desired, and a neurotoxin is needed that can effectively paralyze the target muscle with few undesirable off-target effects.

[0122] A high safety margin is particularly advantageous in a therapeutic method because it represents an increased therapeutic index. In other words, this means that the dosage can be reduced and / or increased dosages can be used without any additional (e.g., adverse) effects compared to alternative clostridial neurotoxin therapeutics. Adverse effects may include systemic toxicity and / or unwanted diffusion to adjacent muscles. The ability to use higher doses of neurotoxin without additional effects is particularly advantageous because higher doses typically result in a longer duration of action of the neurotoxin.

[0123] The efficacy of the chimeric Clostridial neurotoxin is determined by the ability of the neurotoxin to produce a given DAS score, e.g., a DAS score of 2 (ED 50 The potency of a chimeric clostridial neurotoxin can also be expressed as the EC value in a cellular assay measuring SNARE cleavage by the neurotoxin. 50 Dose, e.g., EC in a cellular assay measuring SNAP25 cleavage by the chimeric clostridial neurotoxin 50 It can also be expressed as a dose.

[0124] The duration of action of a chimeric Clostridial neurotoxin can be expressed as the time required to restore the DAS score to 0 after administration of a given dose of neurotoxin, for example, the minimum dose of neurotoxin that results in a DAS score of 4, to the mouse gastrocnemius / soleus muscle complex.

[0125] The chimeric Clostridial neurotoxin may have a safety factor of greater than 7, where the safety factor is the dose of toxin required for a -10% body weight change, measured in pg / mouse, relative to the DAS ED2000, measured in pg / mouse. 50 It is calculated by dividing by ED 50 is the dose required to obtain a DAS score of 2. For example, a chimeric Clostridial neurotoxin can have a safety factor of at least 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50.

[0126] Preferably, the chimeric Clostridial neurotoxin has a safety factor of at least 10 (e.g., a safety factor of 10), more preferably at least 12 or 13 (e.g., 14 to 15). The chimeric Clostridial neurotoxin may have a safety factor of greater than 7, up to 50, e.g., 8 to 45, 10 to 20, or 12 to 15.

[0127] The chimeric clostridial neurotoxins of the present invention preferably have a longer duration of action (e.g., at least a 5%, 10%, 25%, or 50% improvement in one or more symptoms) when compared to BoNT / A (e.g., the two-chain form of SEQ ID NO: 6). The duration of action may be at least 1.25-fold, 1.5-fold, 1.75-fold, 2.0-fold, or 2.25-fold longer. The duration of action of the chimeric clostridial neurotoxin may be between 4.5 and 9 months, or between 6 and 9 months. For example, the duration of action may be at least 4.5, 5.0, 5.5, 6, 6.5, 7.0, 7.5, 8.0, 8.5, or 9.0 months (from onset of action). In certain embodiments, the duration of action may be greater than 9.0 months.

[0128] Thus, in one embodiment, the chimeric clostridial neurotoxin can treat a subject's disorder (e.g., pain or a sensory disorder, preferably pain) for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., the two-chain form of SEQ ID NO: 6). The period of time can be a period of time from administration that is consistent with the duration of action of the chimeric clostridial neurotoxin of the present invention. Thus, the chimeric clostridial neurotoxin can treat a subject's disorder for a period of time from administration that is at least 1.25-fold, 1.5-fold, 1.75-fold, 2.0-fold, or 2.25-fold longer than the treatment period from administration of BoNT / A (e.g., the two-chain form of SEQ ID NO: 6). The chimeric Clostridial neurotoxin may treat a subject's disorder for a period of 4.5 to 9 months, or for a period of 6 to 9 months, e.g., at least 4.5, 5.0, 5.5, 6, 6.5, 7.0, 7.5, 8.0, 8.5, or 9.0 months from administration. In certain embodiments, the chimeric Clostridial neurotoxin may treat a subject's disorder for a period of greater than 9.0 months from administration.

[0129] Accordingly, in one aspect, the present invention provides a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin inhibits release of a mediator (e.g., a pain mediator) from neurons comprising Aδ nerve fibers or C nerve fibers, and wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0130] In a related aspect, the invention provides a chimeric Clostridial neurotoxin for use in a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering the chimeric Clostridial neurotoxin to a subject, wherein the chimeric Clostridial neurotoxin inhibits release of a mediator (e.g., a pain mediator) from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0131] In another related aspect, the present invention provides use of a chimeric Clostridial neurotoxin in the manufacture of a medicament to treat a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., a two-chain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin inhibits release of a mediator (e.g., a pain mediator) from neurons comprising Aδ nerve fibers or C nerve fibers, and wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0132] Thus, in one aspect, the present invention provides a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., the dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0133] In a related aspect, the present invention provides a chimeric Clostridial neurotoxin for use in a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., the dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0134] In another related aspect, the present invention provides use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A (e.g., a di-chain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0135] The disorder is preferably migraine or migraine pain.

[0136] The term "treating a subject's disorder for a longer period of time (e.g., from administration) compared to a subject treated with BoNT / A" can mean that one or more symptoms of the subject's disorder are alleviated for a longer period of time after administration of a chimeric clostridial neurotoxin of the present invention compared to administration of BoNT / A. The duration of action may be at least 1.25-fold, 1.5-fold, 1.75-fold, 2.0-fold, or 2.25-fold longer. The duration of action of the chimeric clostridial neurotoxin may be between 6 and 9 months. For example, the duration of action may be at least 4.5 months, 5.0 months, 5.5 months, 6 months, 6.5 months, 7.0 months, 7.5 months, 8.0 months, 8.5 months, or 9.0 months (from onset of action). In certain embodiments, the duration of action may be greater than 9.0 months. The alleviation may be measured by comparison with an equivalent control patient with comparable symptoms treated with BoNT / A. For a period of time in which the severity of one or more symptoms of a control patient is substantially the same (e.g., the same) as before BoNT / A treatment, a subject treated with a chimeric Clostridial neurotoxin according to the present invention may show at least a 5%, 10%, 25%, or 50% improvement in the equivalent one or more symptoms when compared to the severity of the one or more symptoms before treatment with the chimeric Clostridial neurotoxin.

[0137] In one embodiment, the chimeric Clostridial neurotoxin can treat a subject's disorder (e.g., pain or sensory disorder, preferably pain) with greater efficacy compared to a subject treated with BoNT / A (e.g., the two-chain form of SEQ ID NO: 6).

[0138] Thus, in one aspect, the present invention provides a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject with greater efficacy compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin inhibits release of a mediator (e.g., a pain mediator) from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0139] In a related aspect, the present invention provides a chimeric Clostridial neurotoxin for use in a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject with greater efficacy compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering the chimeric Clostridial neurotoxin to a subject, wherein the chimeric Clostridial neurotoxin inhibits the release of a mediator (e.g., a pain mediator) from neurons comprising Aδ nerve fibers or C nerve fibers, and wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0140] In another related aspect, the present invention provides use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject with greater efficacy compared to a subject treated with BoNT / A (e.g., a two-chain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin inhibits the release of a mediator (e.g., a pain mediator) from a neuron comprising an Aδ nerve fiber or a C nerve fiber, and wherein the chimeric Clostridial neurotoxin binds to a neuron comprising an Aδ nerve fiber or a C nerve fiber, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0141] Thus, in one aspect, the present invention provides a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject with greater efficacy compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0142] In a related aspect, the present invention provides a chimeric Clostridial neurotoxin for use in a method of treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject with greater efficacy compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0143] In another related aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for treating a disorder (e.g., pain or a sensory disorder, preferably pain) in a subject with greater efficacy compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0144] The disorder is preferably migraine or migraine pain.

[0145] The term "treating a subject's disorder with greater efficacy compared to a subject treated with BoNT / A" may mean that one or more symptoms of the subject's disorder are alleviated more after administration of a chimeric clostridial neurotoxin of the present invention compared to administration of BoNT / A. The alleviation may be measured by comparison with an equivalent control patient with equivalent symptoms treated with BoNT / A. At a given time period after administration, a subject treated with a chimeric clostridial neurotoxin according to the present invention may exhibit a reduction in the severity of one or more symptoms of at least 5%, 10%, 25%, or 50% compared to the severity of one or more equivalent symptoms of a control patient at the same time period after administration of BoNT / A (e.g., SEQ ID NO: 6 in a two-chain form). In another embodiment, greater efficacy may mean that the maximum reduction in the severity of one or more symptoms of a subject treated with a chimeric clostridial neurotoxin is greater than the maximum reduction in the severity of one or more equivalent symptoms of a control patient treated with BoNT / A (e.g., SEQ ID NO: 6 in a two-chain form).

[0146] In one embodiment, the chimeric Clostridial neurotoxin can provide greater relief of pain (e.g., migraine pain) in a subject compared to a subject treated with BoNT / A (e.g., the dichain form of SEQ ID NO: 6).

[0147] Thus, in one aspect, the present invention provides a method for reducing pain in a subject compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin inhibits pain mediator release from neurons comprising Aδ nerve fibers or C nerve fibers, and wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0148] In a related aspect, the present invention provides a chimeric Clostridial neurotoxin for use in a method for reducing pain in a subject compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject the chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin inhibits the release of pain mediators from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0149] In another related aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for providing greater pain relief in a subject compared to a subject treated with BoNT / A (e.g., the dichain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin inhibits pain mediator release from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0150] Thus, in one aspect, the present invention provides a method for providing greater pain relief in a subject compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0151] In a related aspect, the present invention provides a chimeric Clostridial neurotoxin for use in a method for reducing pain in a subject compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0152] In another related aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for providing greater relief of pain in a subject compared to a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0153] The pain is preferably migraine pain.

[0154] The term "greater reduction in pain in a subject compared to a subject treated with BoNT / A" may mean that the subject's pain is reduced more after administration of a chimeric clostridial neurotoxin of the present invention compared to administration of BoNT / A. The reduction can be measured by comparison with an equivalent control patient experiencing comparable pain treated with BoNT / A. At a given time period after administration, a subject treated with a chimeric clostridial neurotoxin according to the present invention may exhibit at least a 5%, 10%, 25%, or 50% reduction in pain compared to the equivalent pain severity of a control patient at the same time period after administration of BoNT / A (e.g., SEQ ID NO: 6 in a dichain form). In another embodiment, the maximum reduction in pain in a subject treated with a chimeric clostridial neurotoxin is greater than the maximum reduction in equivalent pain in a control patient treated with BoNT / A (e.g., SEQ ID NO: 6 in a dichain form).

[0155] In one embodiment, the chimeric Clostridial neurotoxin may reduce the amount of a pain mediator (e.g., a migraine pain mediator) in the biological fluids and / or brain of a subject compared to the amount of the same pain mediator in the biological fluids and / or brain of a subject treated with BoNT / A (e.g., the two-chain form of SEQ ID NO: 6).

[0156] Thus, in one aspect, the present invention provides a method for reducing the amount of a pain mediator in a biological fluid and / or brain of a subject compared to the amount of the same pain mediator in the same biological fluid and / or brain of a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin inhibits the release of the pain mediator from neurons comprising Aδ nerve fibers or C nerve fibers, and wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0157] In a related aspect, the invention provides a chimeric Clostridial neurotoxin for use in a method of reducing the amount of a pain mediator in a biological fluid and / or brain of a subject compared to the amount of the same pain mediator in the biological fluid and / or brain of a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject the chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin inhibits the release of the pain mediator from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0158] In another related aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for reducing the amount of a pain mediator in a biological fluid and / or brain of a subject compared to the amount of the same pain mediator in the same biological fluid and / or brain of a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin inhibits the release of the pain mediator from neurons comprising Aδ nerve fibers or C nerve fibers, wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively, and wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0159] Thus, in one aspect, the present invention provides a method of reducing the amount of a pain mediator in a biological fluid and / or brain of a subject compared to the amount of the same pain mediator in the same biological fluid and / or brain of a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0160] In a related aspect, the present invention provides a chimeric Clostridial neurotoxin for use in a method for reducing the amount of a pain mediator in a biological fluid and / or brain of a subject compared to the amount of the same pain mediator in the biological fluid and / or brain of a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), the method comprising administering to the subject a chimeric Clostridial neurotoxin, wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H Cdomain).

[0161] In another related aspect, the present invention provides the use of a chimeric Clostridial neurotoxin in the manufacture of a medicament for reducing the amount of a pain mediator in a biological fluid and / or brain of a subject compared to the amount of the same pain mediator in the biological fluid and / or brain of a subject treated with BoNT / A (e.g., a dichain form of SEQ ID NO: 6), wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and the BoNT / B receptor binding domain (H C domain).

[0162] The term "reducing the amount of pain mediator in a subject's biological fluid and / or brain compared to the amount of the same pain mediator in the same biological fluid and / or brain of a subject treated with BoNT / A" can mean that the amount of pain mediator in a subject is reduced more after administration of a chimeric Clostridial neurotoxin of the present invention compared to administration of BoNT / A. The reduction can be measured by comparing the amount of the same pain mediator in the same biological fluid and / or brain of a comparable control patient treated with BoNT / A. At a given time period after administration, a subject treated with a chimeric Clostridial neurotoxin of the present invention can exhibit at least a 5%, 10%, 25%, or 50% reduction in the amount of pain mediator in the biological fluid and / or brain when compared to the amount of the same pain mediator in the same biological fluid and / or brain of a control patient at the same time period after administration of BoNT / A (e.g., SEQ ID NO: 6 in its dichain form). In another embodiment, the maximum reduction in the amount of pain mediator in biological fluids and / or brains of subjects treated with the chimeric Clostridial neurotoxin is greater than the maximum reduction in the same pain mediator in the same biological fluids and / or brains of control patients treated with BoNT / A (e.g., the dichain form of SEQ ID NO: 6). The pain mediator may be a migraine pain mediator. The pain mediator is preferably CGRP. Preferably, the biological fluid is blood (including a fraction thereof).

[0163] CGRP can be used as a relevant marker for assessing the effectiveness of analgesics, such as painkillers. Therefore, CGRP can be used as a biomarker to determine the suitability of a clostridial neurotoxin for treating pain (e.g., migraine pain). Thus, in one aspect, the present invention provides a method for determining whether a clostridial neurotoxin is suitable for treating pain, the method comprising: (a) comparing the level of CGRP in a first sample with the level of CGRP in a second sample, wherein the first sample is obtained from the subject prior to administration of a clostridial neurotoxin and the second sample is obtained from the same subject after administration of the clostridial neurotoxin; and (b) determining that the Clostridial neurotoxin is suitable for treating pain if the level of CGRP in the second sample is lower than the level of CGRP in the first sample; or (c) determining that the Clostridial neurotoxin is unsuitable for treating pain if the level of CGRP in the second sample is not lower (e.g., higher or the same) than the level of CGRP in the first sample. As used in this context, the term "lower than" preferably means statistically significantly lower, and "not lower" preferably means not statistically significantly different (e.g., the same) or statistically significantly higher.

[0164] The clostridial neurotoxin may be any suitable clostridial neurotoxin known in the art, such as a chimeric clostridial neurotoxin described herein, and may be BoNT / A, BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, BoNT / X, or tetanus neurotoxin (TeNT).

[0165] BoNT / A can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 6. For example, BoNT / A can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 6. Preferably, BoNT / A can comprise (and more preferably consist of) SEQ ID NO: 6.

[0166] BoNT / B can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 7. For example, BoNT / B can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 7. Preferably, BoNT / B can comprise (and more preferably consist of) SEQ ID NO: 7.

[0167] BoNT / C can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 8. For example, BoNT / C can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 8. Preferably, BoNT / C can comprise (and more preferably consist of) SEQ ID NO: 8.

[0168] BoNT / D can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 9. For example, BoNT / D can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 9. Preferably, BoNT / D can comprise (and more preferably consist of) SEQ ID NO: 9.

[0169] BoNT / E can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 10. For example, BoNT / E can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 10. Preferably, BoNT / E can comprise (and more preferably consist of) SEQ ID NO: 10.

[0170] BoNT / F can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 11. For example, BoNT / F can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 11. Preferably, BoNT / F can comprise (and more preferably consist of) SEQ ID NO: 11.

[0171] BoNT / G can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 12. For example, BoNT / G can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 12. Preferably, BoNT / G can comprise (and more preferably consist of) SEQ ID NO: 12.

[0172] BoNT / X can comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 13. For example, BoNT / X can comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 13. Preferably, BoNT / X can comprise (and more preferably consist of) SEQ ID NO: 13.

[0173] TeNT may comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 14. For example, TeNT may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 14. Preferably, TeNT may comprise (and more preferably consist of) SEQ ID NO: 14.

[0174] In one embodiment, prior to using a clostridial neurotoxin in a method for determining whether a clostridial neurotoxin is suitable for treating pain, the clostridial neurotoxin is converted to a di-chain form, e.g., as described herein.

[0175] The first and second samples may be blood samples that have optionally been subjected to one or more processing steps. The first and second samples are preferably comparable (e.g., of the same type and optionally subjected to the same processing steps). The level of CGRP can be measured using any suitable technique, including quantitative Western blotting and / or mass spectrometry.

[0176] BoNT / A light chain, BoNT / A translocation domain, and / or BoNT / BH C The domains include, but are not limited to, modified BoNT / A light chains, BoNT / A translocation domains, and / or BoNT / BH domains described below. C The modified BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain or a derivative thereof may be a modified BoNT / A light chain, a BoNT / A translocation domain, and / or a BoNT / BH domain or a derivative thereof. C The domain or derivative may be a naturally occurring (unmodified) form of the BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH. C The BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH light chain may contain one or more amino acids that are modified compared to the naturally occurring (unmodified) form of the BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH light chain. C The modified BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain may contain one or more inserted amino acids not present in the modified BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain. C The domains may be native (unmodified) BoNT / A light chain, BoNT / A translocation domain, and / or BoNT / BH. C The domain sequences may contain modified amino acid sequences in one or more domains. Such modifications may alter their functional aspects, such as biological activity or biological persistence. Thus, in one embodiment, the BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH C The domain may be a modified BoNT / A light chain, a BoNT / A translocation domain, and / or a BoNT / BH C domain, or modified BoNT / A light chain, BoNT / A translocation domain, and / or BoNT / BH CIt may be a derivative of the domain.

[0177] Modified BoNT / BH C The domain may have one or more modifications that modify binding to target neurons, for example, natural (unmodified) BoNT / BH. C BoNT / BH domains provide higher or lower affinity binding. C Such modifications in the domain alter binding to ganglioside receptors and / or protein receptors on target neurons. C This may involve modifying residues in the ganglioside binding site or in the protein (e.g., synaptotagmin) binding site of the domain. Examples of such modified neurotoxins are described in WO 2006 / 027207 and WO 2006 / 114308, both of which are incorporated by reference in their entireties.

[0178] The modified light chain may have one or more modifications in its amino acid sequence, such as modifications in the substrate binding or catalytic domains, that may alter or modify the SNARE protein specificity of the modified light chain, but which do not catalytically inactivate the light chain. Examples of such modified neurotoxins are described in WO 2010 / 120766 and US 2011 / 0318385, both of which are incorporated by reference in their entireties.

[0179] BoNT / A LH N The domain may correspond to amino acid residues 1 to 872 of SEQ ID NO: 6, or a polypeptide sequence having at least 70% sequence identity thereto. N The domain may correspond to amino acid residues 1 to 872 of SEQ ID NO: 6, or a polypeptide sequence having at least 80%, 90%, or 95% sequence identity thereto. Preferably, the LH domain of BoNT / A N The domain corresponds to amino acid residues 1 to 872 of SEQ ID NO:6.

[0180] BoNT / B H CThe domain may correspond to amino acid residues 860 to 1291 of SEQ ID NO: 7, or a polypeptide sequence having at least 70% sequence identity thereto. C The domain may correspond to amino acid residues 860 to 1291 of SEQ ID NO: 7, or a polypeptide sequence having at least 80%, 90%, or 95% sequence identity thereto. C The domain corresponds to amino acid residues 860 to 1291 of SEQ ID NO:7.

[0181] Preferably, the BoNT / AB chimera is BoNT / A1 LH N Domain and BoNT / B1 H C More preferably, the LH domain N The domain corresponds to BoNT / A amino acid residues 1 to 8721 (SEQ ID NO: 6), C The domain corresponds to amino acid residues 860 to 1291 of BoNT / B1 (SEQ ID NO: 7).

[0182] Most preferably, BoNT / BH C The H domain has the effect of increasing the binding affinity of BoNT / B neurotoxin to human Syt II compared to the native BoNT / B sequence. CC It further comprises a substitution, insertion, indel, or deletion of at least one amino acid residue in the subdomain. CC Suitable amino acid residue substitutions, insertions, indels, or deletions in the subdomains are disclosed in WO 2013 / 180799 and WO 2016 / 154534 (both of which are incorporated herein by reference).

[0183] BoNT / BH CCSuitable amino acid residue substitutions, insertions, indels, or deletions in the subdomains may comprise substitution mutations selected from the group consisting of: V1118M, Y1183M, E1191M, E1191I, E1191Q, E1191T, S1199Y, S1199F, S1199L, S1201V, E1191C, E1191V, E1191L, E1191Y, S1199W, S1199E, S1199H, W1178Y, W1178Q, W1178A, W1178S, Y1183C, Y1183P, and combinations thereof.

[0184] BoNT / BH CC The substitution, insertion, indel, or deletion of a suitable amino acid residue in the subdomain may further comprise a combination of two substitution mutations selected from the group consisting of: E1191M and S1199L, E1191M and S1199Y, E1191M and S1199F, E1191Q and S1199L, E1191Q and S1199Y, E11 91Q and S1199F, E1191M and S1199W, E1191M and W1178Q, E1191C and S1199W, E1191C and S1199Y, E1191C and W1178Q, E1191Q and S1199W, E1191V and S1199W, E1191V and S1199Y, or E1191V and W1178Q.

[0185] BoNT / BH CC Suitable amino acid residue substitutions, insertions, indels, or deletions in the subdomains may also include a combination of the three substitution mutations E1191M, S1199W, and W1178Q.

[0186] Preferably, BoNT / BH CC Suitable amino acid residue substitutions, insertions, indels, or deletions in the subdomain may include a combination of two substitution mutations, E1191M and S1199Y. Such modifications are present in SEQ ID NO: 1 and SEQ ID NO: 4 of the chimeric clostridial neurotoxin. E1191M may correspond to position 1204 and S1199Y to position 1212 of SEQ ID NO: 1. Thus, SEQ ID NO: 1 may include 1204M and 1212Y.

[0187] This modification may be a modification compared to the unmodified BoNT / B set forth as SEQ ID NO:7, where the numbering of amino acid residues is determined by alignment with SEQ ID NO:7. Because the presence of a methionine residue at position 1 of SEQ ID NO:7 is optional (as with the SEQ ID NOs corresponding to the chimeric clostridial neurotoxin polypeptides described herein), one skilled in the art would consider the presence / absence of the methionine residue when determining the amino acid residue numbering. For example, if SEQ ID NO:7 contains a methionine, the position numbering would be as defined above (e.g., E1191 would become E1191 in SEQ ID NO:7). Alternatively, if a methionine is absent in SEQ ID NO:7, the amino acid residue numbering must be corrected by -1 (e.g., E1191 would become E1190 in SEQ ID NO:7). Thus, the first methionine amino acid residue in the polypeptide sequence of a chimeric clostridial neurotoxin may be optional or absent. Similar considerations apply for the presence / absence of methionine at position 1 of other polypeptide sequences described herein, and the skilled artisan will readily determine the correct amino acid residue numbering using techniques routine in the art. Alignments may be performed using any of the methods described herein for determining percent sequence homology and / or sequence identity.

[0188] The term "deletion," as used herein, refers to the removal of one or more amino acid residues of a polypeptide without replacing one or more amino acid residues at the deletion site. Thus, (for example), if one amino acid residue is deleted from a polypeptide sequence having x amino acid residues, the resulting polypeptide will have x-1 amino acid residues.

[0189] The term "indel," as used herein, refers to the deletion of one or more amino acid residues from a polypeptide and the insertion at the site of deletion of a different number of amino acid residues (more or fewer amino acid residues) compared to the number of deleted amino acid residues. Thus, for example, in the case of an indel in which two amino acid residues are deleted from a polypeptide sequence having x amino acid residues, the resulting polypeptide has x-1 amino acid residues, or x+≧1 amino acid residues. Insertions and deletions can be performed sequentially or simultaneously, in any order.

[0190] The term "substitution," as used herein, refers to the replacement of one or more amino acid residues with the same number of amino acid residues at the same positions. Thus, in the case of a substitution of (for example) a polypeptide sequence having x amino acid residues, the resulting polypeptide also has x amino acid residues. Preferably, the substitution is at a single amino acid position.

[0191] The term "insertion," as used herein, refers to the addition of one or more amino acid residues to a polypeptide without deletion of one or more amino acid residues from the polypeptide at the site of insertion. Thus, (for example), if one amino acid residue is inserted into a polypeptide sequence having x amino acid residues, the resulting polypeptide will have x+1 amino acid residues.

[0192] Methods for modifying proteins by substitution, insertion, or deletion of amino acid residues or by indels are known in the art. For example, amino acid modifications can be introduced by modifying a nucleic acid sequence (e.g., a DNA sequence) encoding the polypeptide. This can be achieved by standard molecular cloning techniques, such as site-directed mutagenesis, which uses a short strand of DNA (oligonucleotide) encoding the desired amino acid to replace the original coding sequence using a polymerase enzyme, or by inserting / deleting portions of a gene using various enzymes (e.g., ligase and restriction enzymes). Alternatively, modified gene sequences can be chemically synthesized. Typically, modifications are made by modifying a nucleic acid encoding a naturally occurring clostridial neurotoxin (or a portion thereof), such that the modified clostridial neurotoxin (or a portion thereof) encoded by the nucleic acid contains the modification(s). Alternatively, a nucleic acid encoding a modified clostridial neurotoxin (or a portion thereof) containing the modification(s) can be synthesized.

[0193] A chimeric Clostridial neurotoxin used in the present invention may comprise a polypeptide sequence having at least 70% sequence identity to a polypeptide sequence selected from SEQ ID NOs: 1 to 5. For example, a chimeric Clostridial neurotoxin may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to a polypeptide sequence selected from SEQ ID NOs: 1 to 5. Preferably, a chimeric Clostridial neurotoxin used in the present invention may comprise (more preferably, consist of) a polypeptide sequence selected from SEQ ID NOs: 1 to 5. Of the chimeric Clostridial neurotoxins, SEQ ID NO: 1 is preferred.

[0194] Thus, it is preferred that a chimeric Clostridial neurotoxin comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 1. More preferably, a chimeric Clostridial neurotoxin may comprise a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 1. Most preferably, a chimeric Clostridial neurotoxin for use in the present invention may comprise (and more preferably consist of) SEQ ID NO: 1.

[0195] The two-chain chimeric clostridial neurotoxin of the present invention may comprise an L chain portion of a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 1 to 5, which together constitute the first chain of the two-chain chimeric clostridial neurotoxin, and an H chain portion of a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 1 to 5, which together constitute the second chain of the two-chain chimeric clostridial neurotoxin. N and H C The polypeptide may comprise a domain, wherein the first and second chains are linked to each other by disulfide bonds.

[0196] When cleavage occurs at two or more positions (preferably two positions) within the activation loop of a chimeric clostridial neurotoxin comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 1 to 5, a small fragment of the C-terminal L chain portion of the sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 1 to 5 may be absent from the two-chain chimeric clostridial neurotoxin. In this regard, the sequence of a two-chain chimeric clostridial neurotoxin (e.g., comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 1-5) may differ slightly from the sequence of a corresponding single-chain chimeric clostridial neurotoxin comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 1-5. The subfragment may be 1 to 15 amino acids. In particular, in one embodiment, when Lys-C is used to convert a single-chain chimeric clostridial neurotoxin to a two-chain clostridial neurotoxin, the subfragment of the C-terminal L-chain portion of the absent sequence may be SEQ ID NO: 15 or 16.

[0197] Preferably, the two-chain chimeric Clostridial neurotoxin of the present invention may comprise an L chain portion of a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 1, which together constitute the first chain of the two-chain chimeric Clostridial neurotoxin, and an H chain portion of a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 1, which together constitute the second chain of the two-chain chimeric Clostridial neurotoxin. N and H C The polypeptide may comprise a domain, wherein the first and second chains are linked to each other by disulfide bonds.

[0198] When cleavage occurs at two or more positions (preferably two positions) within the activation loop of a chimeric clostridial neurotoxin comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 1, a small fragment of the C-terminal L-chain portion of the sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 1 may be absent in the two-chain chimeric clostridial neurotoxin. In this regard, the sequence of a two-chain chimeric clostridial neurotoxin (e.g., comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 1) may differ slightly from the sequence of a corresponding single-chain chimeric clostridial neurotoxin comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 1. The subfragment may be 1 to 15 amino acids. In particular, in one embodiment, when Lys-C is used to convert a single-chain chimeric clostridial neurotoxin into a two-chain clostridial neurotoxin, the subfragment of the C-terminal L-chain portion of the absent sequence may be SEQ ID NO: 15 or 16.

[0199] In particularly preferred embodiments, the two-chain chimeric clostridial neurotoxin comprises (or consists of) a light chain comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 17 or 18 (preferably SEQ ID NO: 17) and a heavy chain comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 19, wherein the light chain and the heavy chain are linked to each other by a disulfide bond. More preferably, the two-chain chimeric clostridial neurotoxin comprises (or consists of) a light chain comprising SEQ ID NO: 17 or 18 (preferably SEQ ID NO: 17) and a heavy chain comprising SEQ ID NO: 19, wherein the light chain and the heavy chain are linked to each other by a disulfide bond. More preferably, the two-chain chimeric Clostridial neurotoxin comprises (or consists of) a light chain having SEQ ID NO: 17 and a heavy chain having SEQ ID NO: 19, wherein the light chain and the heavy chain are linked to each other by a disulfide bond. The disulfide bond is preferably formed by and / or between cysteine ​​residue 429 of SEQ ID NO: 17 or 18 and cysteine ​​residue 6 of SEQ ID NO: 19.

[0200] In preferred embodiments, the chimeric Clostridial neurotoxins of the present invention do not include a therapeutic or diagnostic agent (e.g., a nucleic acid, protein, peptide, or small molecule therapeutic, or diagnostic agent) in addition to the light and heavy chains. For example, in one embodiment, the chimeric Clostridial neurotoxin may not include a covalently or non-covalently bound therapeutic or diagnostic agent. Thus, the chimeric Clostridial neurotoxins of the present invention preferably do not function as a delivery vehicle for an additional therapeutic or diagnostic agent.

[0201] In embodiments in which the chimeric clostridial neurotoxins described herein have a tag for purification (eg, a histidine tag) and / or a linker, the tag and / or linker are optional.

[0202] The chimeric clostridial neurotoxins of the present invention may be free of complex proteins present in naturally occurring clostridial neurotoxin complexes.

[0203] The chimeric Clostridial neurotoxins of the present invention can be produced using recombinant nucleic acid technology. Thus, in one embodiment, the chimeric Clostridial neurotoxin (described herein) is a recombinant chimeric Clostridial neurotoxin.

[0204] In one embodiment, a nucleic acid (e.g., DNA) is provided that comprises a nucleic acid sequence encoding a chimeric Clostridial neurotoxin. In one embodiment, the nucleic acid sequence is prepared as part of a DNA vector that includes a promoter and a terminator. The nucleic acid sequence can be selected from any of the nucleic acid sequences described herein.

[0205] In a preferred embodiment, the vector has a promoter selected from: Promoter / Inducer / General induction conditions Tac (hybrid) / IPTG / 0.2mM (0.05 to 2.0mM) AraBAD / L-arabinose / 0.2% (0.002 to 0.4%) T7-lac operator / IPTG / 0.2mM (0.05 to 2.0mM)

[0206] In another preferred embodiment, the vector has a promoter selected from: Promoter / Inducer / General induction conditions Tac (hybrid) / IPTG / 0.2mM (0.05 to 2.0mM) AraBAD / L-arabinose / 0.2% (0.002 to 0.4%) T7-lac operator / IPTG / 0.2mM (0.05 to 2.0mM) T5-lac operator / IPTG / 0.2mM (0.05 to 2.0mM)

[0207] Nucleic acid molecules can be produced using any suitable process known in the art. Thus, nucleic acid molecules can be produced using chemical synthesis techniques. Alternatively, nucleic acid molecules of the present invention can be produced using molecular biology techniques.

[0208] The DNA constructs of the present invention are preferably designed on a computer and then synthesized by conventional DNA synthesis techniques.

[0209] The above nucleic acid sequence information is optionally modified for codon bias depending on the final host cell (eg, E. coli) expression system used.

[0210] As used herein, the terms "nucleotide sequence" and "nucleic acid" are used synonymously. Preferably, the nucleotide sequence is a DNA sequence.

[0211] The chimeric Clostridial neurotoxins of the present invention may exist as a single chain or as two chains. However, chimeric Clostridial neurotoxins may be characterized as having an L chain connected to an H chain (or a component thereof, e.g., H) via a disulfide bond. N Preferably, the protein is present as two chains linked together (domains).

[0212] Production of a single-chain chimeric clostridial neurotoxin having a light chain and a heavy chain can be achieved using a method comprising expressing a nucleic acid encoding the chimeric clostridial neurotoxin in an expression host, lysing the host cells to obtain a host cell homogenate containing the single-chain chimeric clostridial neurotoxin, and isolating the single-chain chimeric clostridial neurotoxin. The single-chain chimeric clostridial neurotoxin described herein can be proteolytically processed using a method comprising contacting the single-chain chimeric clostridial neurotoxin with a protease (e.g., Lys-C) that hydrolyzes the peptide bond in the activation loop of the chimeric clostridial neurotoxin, thereby converting the single-chain chimeric clostridial neurotoxin into the corresponding two-chain chimeric clostridial neurotoxin (e.g., the light chain and the heavy chain are linked to each other by a disulfide bond). A two-chain chimeric clostridial neurotoxin is preferably obtained by such a method.

[0213] Therefore, the chimeric clostridial neurotoxin used in the present invention is preferably a two-chain chimeric clostridial neurotoxin generated from a single-chain BoNT / A, wherein the single-chain BoNT / A comprises or consists of a polypeptide sequence described herein. For example, the chimeric clostridial neurotoxin used in the present invention is preferably a two-chain chimeric clostridial neurotoxin generated from a polypeptide comprising a polypeptide sequence having at least 70% (e.g., at least 80%, 90%, 95%, or 99.9%) sequence identity to SEQ ID NO: 1. Most preferably, the chimeric clostridial neurotoxin used in the present invention is a two-chain chimeric clostridial neurotoxin generated from a polypeptide comprising SEQ ID NO: 1 (more preferably, consisting of SEQ ID NO: 1). Thus, in some embodiments, the chimeric clostridial neurotoxin is a two-chain chimeric clostridial neurotoxin in which the light chain (L chain) is linked to the heavy chain (H chain) via a disulfide bond, obtained by a method comprising contacting a single-chain chimeric clostridial neurotoxin comprising SEQ ID NO: 1 with a protease that hydrolyzes the peptide bond in the activation loop, thereby converting the single-chain chimeric clostridial neurotoxin into a corresponding two-chain chimeric clostridial neurotoxin. In some embodiments, the chimeric clostridial neurotoxin is a two-chain chimeric clostridial neurotoxin in which the L chain is linked to the H chain via a disulfide bond, obtained by a method comprising contacting a single-chain chimeric clostridial neurotoxin consisting of SEQ ID NO: 1 with a protease that hydrolyzes the peptide bond in the activation loop, thereby converting the single-chain chimeric clostridial neurotoxin into a corresponding two-chain chimeric clostridial neurotoxin.

[0214] The term "obtained" as used herein also encompasses "obtained." In one embodiment, the term "obtained" means obtained.

[0215] The protease used to cleave the activation loop is preferably Lys-C. Suitable proteases and methods for cleaving the activation loop to generate a dichain clostridial neurotoxin are taught in WO 2014 / 080206, WO 2014 / 079495, and EP 2677029 A2, which are incorporated herein by reference. Lys-C can cleave the C-terminus of the activation loop to one or more lysine residues present therein. If Lys-C cleaves the activation loop more than once, those skilled in the art will understand that the small peptide in the activation loop of the dichain modified BoNT / A may be absent when compared with the SEQ ID NOs set forth herein (preferably, SEQ ID NOs: 15 or 16 may be absent).

[0216] The term "one or more," as used herein, may mean at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20. In one embodiment, when "one or more" precedes a list, "one or more" may mean every element of the list. Similarly, the term "at least one," as used herein, may mean at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20. In one embodiment, when "at least one" precedes a list, "at least one" may mean every element of the list.

[0217] A "subject," as used herein, may be a mammal, such as a human or other mammal. Preferably, "subject" refers to a human subject.

[0218] The subject of treatment according to the present invention may be a subject who is unsuitable for treatment with a non-chimeric Clostridial neurotoxin. The subject may be a subject who is resistant to treatment with a non-chimeric Clostridial neurotoxin. Resistance may occur due to the subject's development of an immune response to the Clostridial neurotoxin, including the production of anti-Clostridial neurotoxin antibodies. In one embodiment, the subject of treatment according to the present invention may be a subject who is unsuitable for treatment with BoNT / A. The subject may be resistant to treatment with BoNT / A.

[0219] The term "disorder," as used herein, also encompasses "disease." In one embodiment, the disorder is a disease.

[0220] The term "treat" or "treatment," as used herein, encompasses preventative treatment (e.g., preventing the onset of pain) as well as corrective treatment (e.g., treating a subject already suffering from pain). Preferably, "treat" or "treatment," as used herein, refers to corrective treatment.

[0221] In one embodiment, the chimeric Clostridial neurotoxin is administered to a subject who is not experiencing symptoms of pain or a disorder at the time of treatment. Such administration may be suitable for achieving prophylactic treatment of the pain or disorder described herein. In one embodiment, the treatment of migraine (e.g., migraine pain) may be prophylactic treatment. In one embodiment, the chimeric Clostridial neurotoxin is administered to a subject who is not experiencing migraine pain or symptoms of migraine at the time of treatment.

[0222] The term "treat" or "treatment" as used herein relates to a disorder (preferably pain) and / or its symptoms.

[0223] Thus, the chimeric Clostridial neurotoxins of the present invention can be administered to a subject in a therapeutically effective amount or a prophylactically effective amount. Preferably, the chimeric Clostridial neurotoxins of the present invention are administered to a subject in a therapeutically effective amount.

[0224] A "therapeutically effective amount" is any amount of a chimeric Clostridial neurotoxin that, when administered alone or in combination with another agent (preferably alone) to a subject for treating said disorder (preferably pain) (or a symptom thereof), is sufficient to result in treatment of said disorder (preferably pain) or a symptom thereof.

[0225] A "prophylactically effective amount" is any amount of a chimeric Clostridial neurotoxin that, when administered to a subject alone or in combination with other agents (preferably alone), inhibits or delays the onset or recurrence of a disorder (preferably pain) (or symptoms thereof). In some embodiments, a prophylactically effective amount completely prevents the onset or recurrence of a disorder (preferably pain). "Inhibiting" onset means either reducing the likelihood of onset (preferably pain) (or symptoms thereof), preventing the amplitude of the peak effect of the disorder (preferably pain), and / or preventing onset completely.

[0226] The chimeric clostridial neurotoxins can treat pain without treating the underlying disorder that causes the pain.

[0227] In addition to treating pain, the chimeric clostridial neurotoxin may treat one or more additional symptoms of a disorder. In one embodiment, the chimeric clostridial neurotoxin may treat one or more additional symptoms associated with neuronal secretion, e.g., release of a mediator (e.g., a pain mediator), as described herein. For example, CGRP may be involved in many symptoms associated with migraine, such as photophobia. Thus, treating migraine or migraine pain according to the present invention may also treat one or more additional symptoms of migraine, such as photophobia.

[0228] The chimeric Clostridial neurotoxins of the present invention can be formulated in any suitable manner, for example, for administration to a subject as part of a pharmaceutical composition, which can include a chimeric Clostridial neurotoxin of the present invention, a pharmaceutically acceptable carrier, excipient, adjuvant, propellant, and / or salt.

[0229] The chimeric Clostridial neurotoxins of the present invention can be formulated for oral, parenteral, continuous infusion, inhalation, or topical application. Compositions suitable for injection may be in the form of a solution, suspension, or emulsion, or a dry powder that is dissolved or suspended in a suitable solvent before use.

[0230] In one aspect, the invention provides a unit dosage form of a chimeric Clostridial neurotoxin for the treatment of pain, the unit dosage form comprising: a. 0.2 units up to 707 units of a chimeric Clostridial neurotoxin, where 1 unit is the calculated median lethal dose (LD) in mice. 50 ) an amount of chimeric Clostridial neurotoxin equivalent to b. 5 pg to 17,000 pg of a chimeric Clostridial neurotoxin; and c. Optionally, pharmaceutically acceptable carriers, excipients, adjuvants, and / or salts.

[0231] Preferably, the chimeric Clostridial neurotoxin in unit dosage form comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 1. For example, a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 1. Most preferably, the chimeric Clostridial neurotoxin may comprise (and more preferably consist of) SEQ ID NO: 1.

[0232] A unit dosage form for treating pain can contain from 0.2 units up to 707 units of a chimeric Clostridial neurotoxin. The upper end of the range can be 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, or 100 units of a chimeric Clostridial neurotoxin, preferably 666 units. The lower end of the range can be 40, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, or 700 units of a chimeric Clostridial neurotoxin, preferably 42 units or 31 units. The lower end of the range can be greater than 125 units. Preferably, the unit dosage form contains 31 to 707 units of the chimeric Clostridial neurotoxin. More preferably, the unit dosage form contains 42 to 666 units of the chimeric Clostridial neurotoxin, e.g., 200 to 400 units of the chimeric Clostridial neurotoxin, or 41 to 229 units, e.g., 83 to 188 units, 83 to 125 units (e.g., 104 units), or 145 to 188 units of the chimeric Clostridial neurotoxin. Preferably, the unit dosage form contains 166 units of the chimeric Clostridial neurotoxin. A unit dosage form can contain 47 to 707 units of a chimeric Clostridial neurotoxin, e.g., 187 to 282 units of a chimeric Clostridial neurotoxin, or 47 to 258 units, e.g., 94 to 211 units, 94 to 141 units (e.g., 117 units), or 164 to 211 units of a chimeric Clostridial neurotoxin. A unit dosage form can contain 188 units of a chimeric Clostridial neurotoxin.

[0233] A unit dosage form for treating pain can contain 5 pg to 17,000 pg of chimeric Clostridial neurotoxin, with the upper end of the range being 16,500, 15,500, 14,500, 13,500, 12,500, 11,500, 10,500, 9,500, 8,500, 7,500, 6,500, 5,500, 4,500, 3,500, 2,500, 1,500, or 500 pg of chimeric Clostridial neurotoxin, preferably with an upper end of 16,000 pg. The lower end of the range may be 750, 850, 950, 1000, 1500, 2000, 2,500, 3,000, 3,500, 4,000, 4,500, or 5,000 pg of chimeric Clostridial neurotoxin, preferably, the lower end is 1000 pg or 750 pg. The lower end of the range may be greater than 3,000 pg. Preferably, the unit dosage form contains 750 pg to 17,000 pg of chimeric Clostridial neurotoxin. More preferably, the unit dosage form contains 1,000 pg to 16,000 pg of chimeric Clostridial neurotoxin, e.g., 4,000 pg to 6,000 pg of chimeric Clostridial neurotoxin, or 1,000 to 5,500 pg, e.g., 2,000 pg to 4,500 pg, 2,000 pg to 3,000 pg (e.g., 2,500 pg), or 3,500 to 4,500 pg of chimeric Clostridial neurotoxin. Preferably, the unit dosage form contains 4,000 pg of chimeric Clostridial neurotoxin.

[0234] In some embodiments, the unit dosage form for treating pain may be for treating headache (e.g., migraine pain) or migraine headache and may include: a. 42 to a maximum of 258 units (e.g., 42 to 229 units) of a chimeric Clostridial neurotoxin, where 1 unit is the calculated median lethal dose (LD) in mice. 50 ) an amount of chimeric Clostridial neurotoxin equivalent to b. 1,000 pg to 5,500 pg of a chimeric Clostridial neurotoxin; and c. Optionally, pharmaceutically acceptable carriers, excipients, adjuvants, and / or salts.

[0235] The potency of chimeric Clostridial neurotoxins for use according to the present invention was determined by mouse LD 50 In said assay, one unit corresponds to the calculated median lethal dose (LD) in mice. 50 ) is defined as the amount of chimeric Clostridial neurotoxin equivalent to the median intraperitoneal lethal dose calculated in mice.

[0236] The amount of the chimeric Clostridial neurotoxin corresponding to 1 unit in the assay may be 20 to 24.04 pg, for example, 21.3 pg or 24.04 pg. Preferably, the amount of the chimeric Clostridial neurotoxin corresponding to 1 unit in the assay may be 24.04 pg.

[0237] When referring to units here, the units are preferably LD 50 It is a unit.

[0238] In another aspect, the present invention provides a unit dosage form for treating headache (e.g., migraine pain) or migraine headache, comprising: a. 42 to a maximum of 258 units (e.g., 42 to 229 units) of a chimeric Clostridial neurotoxin, where 1 unit is the calculated median lethal dose (LD) in mice. 50 ) an amount of chimeric Clostridial neurotoxin equivalent to b. 1,000 pg to 5,500 pg of a chimeric Clostridial neurotoxin; and c. Optionally, pharmaceutically acceptable carriers, excipients, adjuvants, and / or salts.

[0239] Preferably, the chimeric Clostridial neurotoxin in unit dosage form comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 1. For example, a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 1. Most preferably, the chimeric Clostridial neurotoxin may comprise (and more preferably consist of) SEQ ID NO: 1.

[0240] A unit dosage form for treating headache (e.g., migraine pain) or migraine headache can be 42 units to 229 units. The upper limit of the unit dosage form can be 225, 220, 215, 210, 205, 200, 190, 180, 170, 160, 150, 125, 100, or 83 units of chimeric Clostridial neurotoxin, preferably 212 units and more preferably 208 units. The lower limit of the unit dosage form can be 46, 50, 55, 60, 65, 70, 75, 80, 90, 100, 110, 120, 130, 140, 150, 160, or 166 units of chimeric Clostridial neurotoxin, preferably 58 units and more preferably 62 units. The lower limit of the range can be greater than 125 units. A unit dosage form may contain 58 to 212 units (e.g., 62 to 208 units), 83 to 212 units, 125 to 212 units, or 125 to 166 units of a chimeric Clostridial neurotoxin. A unit dosage form may contain more than 125 units, up to 229 units of a chimeric Clostridial neurotoxin. Preferably, a unit dosage form contains 83 to 188 units, 83 to 125 units (e.g., 104 units), or 145 to 188 units of a chimeric Clostridial neurotoxin. Preferably, a unit dosage form contains 166 units of a chimeric Clostridial neurotoxin. A unit dosage form may contain 47 to 258 units of a chimeric Clostridial neurotoxin, for example, 94 to 211 units, 94 to 141 units (e.g., 117 units), or 164 to 211 units of a chimeric Clostridial neurotoxin. A unit dosage form can contain 188 units of a chimeric clostridial neurotoxin.

[0241] A unit dosage form for treating headache (e.g., migraine pain) or migraine headache may be 1,000 pg to 5,500 pg. The upper limit of the unit dosage form may be 5,250, 5,200, 5,100, 5,000, 4,500, 4,000, 3,500, 3,000, 2,500, or 2,000 pg of chimeric Clostridial neurotoxin, preferably the upper limit is 5,100 pg, and more preferably 5,000 pg. The lower limit of the unit dosage form may be 1,100, 1,200, 1,250, 1,300, 1,350, 1,400, 1,450, 1,500, 2,000, 2,500, 3,000, 3,500, or 4,000 pg of chimeric Clostridial neurotoxin, preferably the lower limit is 1,400 pg, more preferably 1,500 pg. The lower limit of the range may be greater than 3,000 pg. The unit dosage form may contain 1,400 pg to 5,100 pg, 2,000 pg to 5,100 pg, 3,000 to 5,100 pg, or 3,000 to 4,000 pg of chimeric Clostridial neurotoxin. The unit dosage form may contain more than 3,000 pg, up to 5,500 pg, of the chimeric Clostridial neurotoxin. Preferably, the unit dosage form contains 2,000 pg to 4,500 pg, 2,000 pg to 3,000 pg (e.g., 2,500 pg), or 3,500 to 4,500 pg of the chimeric Clostridial neurotoxin. Preferably, the unit dosage form contains 4,000 pg of the chimeric Clostridial neurotoxin.

[0242] In the case of a chimeric clostridial neurotoxin that is delivered locally, the chimeric clostridial neurotoxin can be formulated as a cream (eg, for topical application) or for subcutaneous injection.

[0243] Local delivery means may include aerosols or other sprays (e.g., nebulizers). In this regard, aerosol formulations of chimeric Clostridial neurotoxins allow for delivery to the lungs and / or other nasal and / or bronchial or airway passages.

[0244] The chimeric Clostridial neurotoxin can be administered to the face, neck, and / or skull of a subject. For example, the chimeric Clostridial neurotoxin can be administered to the muscle and / or skin components thereof. The chimeric Clostridial neurotoxin can be administered to two or more sites among the face, neck, and skull, and preferably to the face, neck, and skull. In particular, the chimeric Clostridial neurotoxin can be administered to the face, neck, and / or skull regions of a subject.

[0245] The chimeric clostridial neurotoxins of the invention can be administered to a subject by intrathecal or epidural injection in the spinal column at the level of the spinal segment involved in innervating the affected organ.

[0246] The route of administration may be via laparoscopy and / or local injection. In one embodiment, the chimeric Clostridial neurotoxin of the present invention is administered at or near the treatment site, preferably at the treatment site. For example, the chimeric Clostridial neurotoxin may be administered intrathecally or intraspinally. In one embodiment, the route of administration of the chimeric Clostridial neurotoxin of the present invention may be intraspinal and / or intrathecal.

[0247] In one embodiment, the chimeric Clostridial neurotoxin of the present invention can be administered peripherally, hi one embodiment, the chimeric Clostridial neurotoxin can be administered subcutaneously.

[0248] The chimeric Clostridial neurotoxin of the present invention can be administered via injection. The chimeric Clostridial neurotoxin can be administered at least 5, 10, 15, 20, 25, or 30 injection sites per treatment session. The chimeric Clostridial neurotoxin can be administered by injection at a maximum of 50, 45, 40, 35, 30, 25, or 20 injection sites per treatment session. The chimeric Clostridial neurotoxin can be administered by injection at a maximum of 20 or 15 injection sites per treatment session. Preferably, the chimeric Clostridial neurotoxin can be administered by injection at a maximum of 10 injection sites per treatment session, for example, a maximum of 9, 8, 7, 6, 5, 4, 3, or 2 injection sites. In one embodiment, the chimeric clostridial neurotoxin may be administered to 1 to 40, 5 to 40, 8 to 38, 30 to 40 (e.g., 35), or 15 to 25 (e.g., 20) injection sites per treatment session. In one embodiment, the chimeric clostridial neurotoxin may be administered to 1 to 10, 3 to 10, 5 to 10, or 7 to 10 injection sites per treatment. In one embodiment, the chimeric clostridial neurotoxin may be administered to 25 to 35 (e.g., 31) injection sites per treatment session. Preferably, the chimeric clostridial neurotoxin may be administered to 25 to 30 (e.g., 28) injection sites per treatment session.

[0249] The chimeric Clostridial neurotoxin of the present invention can be administered intradermally, for example, by intradermal injection. The chimeric Clostridial neurotoxin can be administered by intradermal injection to at least 5, 10, 15, 20, 25, or 30 injection sites per treatment session. The chimeric Clostridial neurotoxin can be administered by intradermal injection to up to 50, 45, 40, 35, 30, 25, or 20 injection sites per treatment session. The chimeric Clostridial neurotoxin can be administered by intradermal injection to up to 20 or 15 injection sites per treatment session. Preferably, the chimeric Clostridial neurotoxin can be administered by intradermal injection to up to 10 injection sites per treatment session, for example, up to 9, 8, 7, 6, 5, 4, 3, or 2 injection sites per treatment session. In one embodiment, the chimeric clostridial neurotoxin may be administered at 1 to 40, 5 to 40, 8 to 38, 30 to 40 (e.g., 35), or 15 to 25 (e.g., 20) injection sites per treatment session. In one embodiment, the chimeric clostridial neurotoxin may be administered at 1 to 10, 3 to 10, 5 to 10, or 7 to 10 injection sites per treatment. In one embodiment, the chimeric clostridial neurotoxin may be administered at 25 to 35 (e.g., 31) injection sites per treatment session. Preferably, the chimeric clostridial neurotoxin may be administered at 25 to 30 (e.g., 28) injection sites per treatment session. Intradermal injections may be administered into a muscle region, such as the muscles described herein. In one embodiment, the intradermal injections may be administered into the skin overlying the muscle.

[0250] Most preferably, the chimeric Clostridial neurotoxin can be administered intramuscularly, for example, by intramuscular injection. The particular muscle into which the chimeric Clostridial neurotoxin is administered will depend on the nature and location of the disorder (preferably pain) being treated.

[0251] The chimeric Clostridial neurotoxin can be administered to one or more muscles of a subject selected from the following: frontalis muscle, corrugator muscle (e.g., corrugator supercilii muscle), procerus muscle (e.g., procerus procerus muscle), occipitalis muscle, temporalis muscle, trapezius muscle, masseter muscle, nasal muscle, orbicularis oculi muscle, cervical paraspinal muscles, temporalis fascia, superior auricular muscle, anterior auricular muscle, posterior auricular muscle, sternocleidomastoid muscle, platysma muscle, dilator naris anterior muscle, dilator naris posterior muscle, depressor septum nasalis muscle, mentalis muscle, orbicularis oris muscle, zygomaticus muscle, laughing muscle, buccinator muscle, occipitofrontalis muscle, levator labii superioris muscle, depressor labii inferioris muscle, depressor anguli oris muscle, thyrohyoid muscle, omohyoid muscle, sternohyoid muscle, splenius cervix muscle, splenius capitis muscle, semispinalis cervix muscle, semispinalis capitis muscle, levator scapulae muscle, digastric muscle, or scalene muscle.

[0252] For example, a chimeric Clostridial neurotoxin can be administered to one or more muscles of a subject selected from the following: frontalis muscle, corrugator muscle, procerus muscle (e.g., procerus procerus), occipitalis muscle, temporalis muscle, trapezius muscle, masseter muscle, nasal muscle, orbicularis oculi muscle, cervical paraspinal muscles, temporalis fascia, superior auricular muscle, anterior auricular muscle, posterior auricular muscle, sternocleidomastoid muscle, platysma muscle, dilator naris anterior muscle, dilator naris posterior muscle, depressor septum nasalis muscle, mentalis muscle, orbicularis oris muscle, zygomaticus muscle, laughing muscle, buccinator muscle, occipitofrontalis muscle, levator labii superioris muscle, depressor labii inferioris muscle, depressor anguli oris muscle, thyrohyoid muscle, omohyoid muscle, sternohyoid muscle, splenius cervix muscle, levator scapulae muscle, digastric muscle, and scalene muscle.

[0253] If the disorder is headache (e.g., migraine pain) or migraine, the invention can include administering a chimeric Clostridial neurotoxin to: the frontalis muscle, the corrugator muscle (e.g., the corrugator supercilii muscle), the procerus muscle (e.g., the procerus muscle), the occipitalis muscle, the temporalis muscle, the trapezius muscle, the masseter muscle, the nasal muscles, the orbicularis oculi muscle, the cervical paraspinal muscles, the temporal fascia, the superior auricular muscle, the anterior auricular muscle, the posterior auricular muscle, the sternocleidomastoid muscle, the platysma muscle, the dilator naris anterior muscle, the dilator naris posterior muscle, the depressor septum nasalis muscle, the mentalis muscle, the orbicularis oris muscle, the zygomaticus muscle, the laughing muscle, the buccinator muscle, the occipitofrontalis muscle, the levator labii superior muscle, the depressor labii inferior muscle, the depressor anguli oris muscle, the thyrohyoid muscle, the omohyoid muscle, the sternohyoid muscle, the splenius cervix muscle, the levator scapulae muscle, the digastric muscle, and the scalene muscle. If the disorder is headache (e.g., migraine pain) or migraine, the chimeric Clostridial neurotoxin can be administered to one or more muscles of the subject selected from the following: frontalis muscle, corrugator muscle (e.g., corrugator supercilii muscle), procerus muscle (e.g., procerus noris muscle), occipitalis muscle, temporalis muscle, trapezius muscle, masseter muscle, nasal muscle, orbicularis oculi muscle, cervical paraspinal muscles, temporalis fascia, superior auricular muscle, anterior auricular muscle, posterior auricular muscle, sternocleidomastoid muscle, platysma muscle, dilator naris anterior muscle, dilator naris posterior muscle, depressor septum nasalis muscle, mentalis muscle, orbicularis oris muscle, zygomaticus muscle, laugher muscle, buccinator muscle, occipitofrontalis muscle, levator labii superioris muscle, depressor labii inferioris muscle, depressor anguli oris muscle, thyrohyoid muscle, omohyoid muscle, sternohyoid muscle, splenius cervix muscle, levator scapulae muscle, digastric muscle, and scalene muscle. Preferably, the chimeric Clostridial neurotoxin is administered to one or more of the following muscles: procerus, corrugator supercilii, masseter, temporalis, occipitalis, and trapezius. If there are two versions of the same muscle (e.g., two occipitalis muscles), the chimeric Clostridial neurotoxin can be administered to one or both of the muscles according to the needs of the subject. Preferably, the chimeric Clostridial neurotoxin is administered to both of the muscles.

[0254] The chimeric Clostridial neurotoxin can be administered intramuscularly, for example, by intramuscular injection. The particular muscle into which the chimeric Clostridial neurotoxin is administered will depend on the nature and location of the disorder (preferably pain) being treated. If the disorder is headache (e.g., migraine pain) or migraine, the invention can include administering a chimeric Clostridial neurotoxin to: the frontalis muscle, the corrugator muscle (e.g., the corrugator supercilii muscle), the procerus muscle (e.g., the procerus muscle), the occipitalis muscle, the temporalis muscle, the trapezius muscle, the masseter muscle, the nasal muscle, the orbicularis oculi muscle, the cervical paraspinal muscles, the temporalis fascia, the superior auricular muscle, the anterior auricular muscle, the posterior auricular muscle, the sternocleidomastoid muscle, the platysma muscle, the dilator naris anterior muscle, the dilator naris posterior muscle, the depressor septum nasalis muscle, the mentalis muscle, the orbicularis oris muscle, the zygomaticus muscle, the lolus muscle, the buccinator muscle, the occipitofrontalis muscle, the levator labii superioris muscle, the depressor labii inferioris muscle, the depressor anguli oris muscle, the thyrohyoid muscle, the omohyoid muscle, the sternohyoid muscle, the splenius cervix muscle, the splenius capitis muscle, the semispinalis cervix muscle, the semispinalis capitis muscle, the levator scapulae muscle, the digastric muscle, or the scalene muscle. If the disorder is headache (e.g., migraine pain) or migraine, the chimeric Clostridial neurotoxin can be administered to one or more muscles of the subject selected from the following: frontalis muscle, corrugator muscle (e.g., corrugator supercilii muscle), procerus muscle (e.g., procerus noris muscle), occipitalis muscle, temporalis muscle, trapezius muscle, masseter muscle, nasal muscle, orbicularis oculi muscle, cervical paraspinal muscles, temporalis fascia, superior auricular muscle, anterior auricular muscle, posterior auricular muscle, sternocleidomastoid muscle, platysma muscle, dilator naris anterior muscle, dilator naris posterior muscle, depressor septum nasalis muscle, mentalis muscle, orbicularis oris muscle, zygomaticus muscle, lolis muscle, buccinator muscle, occipitofrontalis muscle, levator labii superioris muscle, depressor labii inferior muscle, depressor anguli oris muscle, thyrohyoid muscle, omohyoid muscle, sternohyoid muscle, splenius cervix muscle, splenius capitis muscle, semispinalis cervix muscle, semispinalis capitis muscle, levator scapulae muscle, digastric muscle, and scalene muscle. Preferably, the chimeric Clostridial neurotoxin is administered to one or more of the following muscles: procerus, corrugator supercilii, masseter, temporalis, occipitalis, and trapezius. If there are two versions of the same muscle (e.g., two occipitalis muscles), the chimeric Clostridial neurotoxin can be administered to one or both of the muscles according to the needs of the subject. Preferably, the chimeric Clostridial neurotoxin is administered to both of the muscles.

[0255] The invention can include administering a chimeric Clostridial neurotoxin to at least one of the following: the frontalis muscle, the corrugator (e.g., corrugator supercilii), the procerus muscle (e.g., procerus nostril), the occipitalis muscle (e.g., upper occipitalis or lower occipitalis), the temporalis muscle, the trapezius muscle (e.g., upper trapezius, middle trapezius, or lower trapezius), the masseter muscle, the nasal muscle, the orbicularis oculi muscle, the cervical paraspinal muscles, temporalis fascia, superior auricular muscle, anterior auricular muscle, posterior auricular muscle, sternocleidomastoid muscle, platysma muscle, dilator naris anterior muscle, dilator naris posterior muscle, depressor septum naris muscle, mentalis muscle, orbicularis oris muscle, zygomaticus muscle, laughing muscle, buccinator muscle, occipitofrontalis muscle, levator labii superioris muscle, depressor labii inferioris muscle, depressor anguli oris muscle, thyrohyoid muscle, omohyoid muscle, sternohyoid muscle, splenius cervix muscle, splenius capitis muscle, semispinalis cervix muscle, semispinalis capitis muscle, levator scapulae muscle, digastric muscle, or scalene muscle. Preferably, the present invention may comprise administering a chimeric Clostridial neurotoxin to at least one of the frontalis muscle, corrugator (e.g., corrugator supercilii) muscle, nasal muscle, orbicularis oculi muscle, temporalis muscle, occipitalis muscle, or trapezius muscle. More preferably, the present invention may include administering a chimeric Clostridial neurotoxin to the frontalis muscle, corrugator (e.g., corrugator supercilii), nasal muscle, orbicularis oculi, temporalis, occipitalis, and trapezius muscles. When there are two versions of the same muscle (e.g., two occipitalis muscles), the chimeric Clostridial neurotoxin can be administered to one or both of the muscles depending on the needs of the subject. Preferably, the chimeric Clostridial neurotoxin is administered to both of the muscles. The administration may be particularly relevant to the treatment of headache (e.g., migraine pain) or migraine headache.

[0256] When the pain is arthritic pain, the chimeric Clostridial neurotoxin can be administered to one or more muscles in the subject's hand, wrist, knee, and / or foot, depending on, for example, the location of the arthritis and / or arthritic pain. When administered to the subject's hand, wrist, knee, or foot, it can be administered unilaterally (e.g., when the arthritis or arthritic pain is in only one hand, wrist, knee, and / or foot) or bilaterally (e.g., when the arthritis or arthritic pain is in both hands, wrists, knees, and / or feet). The chimeric Clostridial neurotoxin may be administered to one or more muscles of a subject's hand selected from the flexor pollicis brevis, palmar interosseus, abductor pollicis brevis, flexor pollicis brevis, abductor pollicis brevis, opponens pollicis, dorsal interosseus, abductor digiti minimi, flexor digiti minimi, and opponens digiti minimi (preferably one or more selected from the flexor pollicis brevis, palmar interosseus, abductor pollicis brevis, flexor pollicis brevis, and abductor pollicis brevis). The chimeric Clostridial neurotoxin may be administered to one or more muscles of a subject's wrist selected from the extensor pollicis brevis, abductor pollicis longus, extensor digitorum minimi, extensor carpi ulnaris, flexor carpi ulnaris, extensor digitorum radialis, and brachioradialis. The chimeric Clostridial neurotoxin may be administered to one or more muscles of the subject's knee selected from the following: sartorius, vastus medialis, vastus lateralis, gastrocnemius, plantaris, semimembranosus, peroneus longus, gastrocnemius, tibialis anterior, rectus femoris, peroneus longus, iliopsoas, pubococcus, adductor longus, adductor magnus, gracilis, biceps femoris, soleus, soleus, extensor digitorum longus, extensor hallucis longus, peroneus brevis, and flexor digitorum longus (preferably one or more selected from the sartorius, vastus medialis, vastus lateralis, gastrocnemius, plantaris, semimembranosus, peroneus longus, gastrocnemius, tibialis anterior, rectus femoris, and peroneus longus). The chimeric Clostridial neurotoxin can be administered to one or more muscles of the subject's foot selected from the extensor digitorum brevis and the extensor hallucis brevis muscles.

[0257] The chimeric Clostridial neurotoxin may be administered by intramuscular injection to at least 5, 10, 15, 20, 25, or 30 injection sites per treatment session. The chimeric Clostridial neurotoxin may be administered by intramuscular injection to up to 50, 45, 40, 35, 30, 25, or 20 injection sites per treatment session. The chimeric Clostridial neurotoxin may be administered by intramuscular injection to up to 20 or 15 injection sites per treatment session. Preferably, the chimeric Clostridial neurotoxin may be administered by intramuscular injection to up to 10 injection sites per treatment session, for example, up to 9, 8, 7, 6, 5, 4, 3, or 2 injection sites. In one embodiment, the chimeric clostridial neurotoxin may be administered to 1 to 40, 5 to 40, 8 to 38, 30 to 40 (e.g., 35), or 15 to 25 (e.g., 20) injection sites per treatment session. In one embodiment, the chimeric clostridial neurotoxin may be administered to 1 to 10, 3 to 10, 5 to 10, or 7 to 10 injection sites per treatment. In one embodiment, the chimeric clostridial neurotoxin may be administered to 25 to 35 (e.g., 31) injection sites per treatment session. Preferably, the chimeric clostridial neurotoxin may be administered to 25 to 30 (e.g., 28) injection sites per treatment session.

[0258] The chimeric clostridial neurotoxin can be administered in a unit dose per injection (eg, per injection site).

[0259] The chimeric clostridial neurotoxin can be administered by intraneural, perineural, or periganglionic administration.

[0260] The chimeric clostridial neurotoxin can be administered to the upper cervical roots via the trigeminal nerve, trigeminal ganglion, pterygopalatine ganglion, Gasserian ganglion, nerve intermedius, glossopharyngeal, vagus nerve, otic ganglion, and / or occipital nerve. Preferably, the chimeric clostridial neurotoxin is administered to the trigeminal nerve, trigeminal ganglion, and / or pterygopalatine ganglion.

[0261] The chimeric clostridial neurotoxin can be administered intra-articularly. The chimeric clostridial neurotoxin can be administered intramuscularly near a joint and / or intradermally.

[0262] The chimeric clostridial neurotoxin can be administered by perivascular administration.

[0263] The dosage ranges for administering the chimeric Clostridial neurotoxins of the present invention are those that provide the desired therapeutic and / or prophylactic effect.

[0264] Liquid dosage forms are typically prepared using a chimeric Clostridial neurotoxin and a sterile, pyrogen-free solvent. The chimeric Clostridial neurotoxin may be dissolved or suspended in the solvent, depending on the solvent and concentration used. To prepare a solution, the chimeric Clostridial neurotoxin is dissolved in the solvent, sodium chloride is added, if necessary, to make the solution isotonic, and the solution is sterilized by filtration through a sterile filter using aseptic techniques, after which the solution is filled into a suitable sterile vial or ampoule and sealed. Alternatively, if the solution is sufficiently stable, the solution in the sealed container may be sterilized by autoclaving. Advantageously, additives such as buffers, solubilizers, stabilizers, preservatives or bactericides, suspending or emulsifying agents, and / or local anesthetics may be dissolved in the solvent.

[0265] Using aseptic technique in a sterile area, pre-sterilized ingredients may be filled into sterile containers to prepare dry powders that are dissolved or suspended in a suitable solvent before use. Alternatively, the ingredients may be dissolved in a suitable container using aseptic technique in a sterile area. The product is then lyophilized and the container is aseptically sealed.

[0266] Parenteral suspensions suitable for the routes of administration described herein are prepared in substantially the same manner, except that the sterile ingredients are suspended in a sterile solvent rather than dissolved, and sterilization cannot be achieved by filtration. The ingredients may be separated under sterile conditions, or may be sterilized after separation, for example, by gamma irradiation.

[0267] Advantageously, the composition contains a suspending agent, such as polyvinylpyrrolidone, to promote uniform distribution of the ingredients.

[0268] Administration according to the present invention may utilize a variety of delivery techniques, including microparticle encapsulation or high-pressure aerosol impaction.

[0269] Unlike conventional clostridial neurotoxins (e.g., native BoNT / A), the chimeric clostridial neurotoxins of the present invention have an improved safety profile and / or improved activity, as evidenced, for example, by an improved safety ratio compared to conventional neurotoxins (see WO 2017 / 191315 A1 for further details). In this regard, the chimeric clostridial neurotoxins can be administered at low doses while ensuring therapeutic efficacy, and at high doses without causing undesirable toxicity-related side effects. Thus, the present invention provides a wide range of suitable dosages for treating disorders (preferably pain).

[0270] For the convenience of the physician, the chimeric Clostridial neurotoxin may be administered in unit doses. The unit dose may be administered at a single site, or less than a unit dose may be administered at each administration site (e.g., when there is more than one administration site and the dose is divided (equally or unequally) among the sites). In one embodiment, in practicing the present invention, a single unit dose may be administered per muscle and / or neuron to be treated.

[0271] In one embodiment, at least one unit dose can be administered to a muscle and / or neuron when practicing the present invention, for example, 1 to 20, 1 to 10, 1 to 7, or 1 to 5 unit doses can be administered to a muscle and / or neuron when practicing the present invention.

[0272] In one embodiment, at least 0.25, 0.5, 1, or 2 unit doses may be administered per injection (e.g., per injection site). For example, 0.25, 0.5, 1, or 2 unit doses may be administered per injection (e.g., per injection site). Preferably, 1 unit dose may be administered per injection (e.g., per injection site).

[0273] When a unit dose (or a portion or portions thereof) is administered, it can mean that substantially all of the unit dose (or a portion or multiple thereof) is administered. For example, a residual amount (e.g., up to 1%, 0.1%, or 0.01%) of the unit dose (or a portion or portions thereof) may remain in the vial after the chimeric Clostridial neurotoxin has been removed (e.g., the vial into which the chimeric Clostridial neurotoxin has been reconstituted). However, preferably, all of the unit dose (or a portion or portions thereof) is administered (e.g., into one or more injection sites).

[0274] A suitable unit dose may be between 5 pg and 17,000 pg of chimeric Clostridial neurotoxin. The upper end of the unit dose range may be 16,500, 15,500, 14,500, 13,500, 12,500, 11,500, 10,500, 9,500, 8,500, 7,500, 6,500, 5,500, 4,500, 3,500, 2,500, 1,500, or 500 pg of chimeric Clostridial neurotoxin, preferably with an upper limit of 16,000 pg. The lower limit of the unit dose range may be 10, 20, 30, 50, 100, 200, 250, 350, 450, 550, 650, 750, 850, 950, 1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, or 5,000 pg of chimeric Clostridial neurotoxin, preferably 1,000 pg or 750 pg. The lower limit of the range may be greater than 3,000 pg. Preferably, the unit dose is 750 pg to 17,000 pg of chimeric Clostridial neurotoxin. The unit dose of chimeric Clostridial neurotoxin is 3,640 pg to 17,000 pg. More preferably, the unit dose of the chimeric Clostridial neurotoxin is 1,000 pg to 16,000 pg of the chimeric Clostridial neurotoxin, for example, 4,000 pg to 6,000 pg of the chimeric Clostridial neurotoxin, or 1,000 to 5,500 pg, for example, 2,000 pg to 4,500 pg, 2,000 pg to 3,000 pg (e.g., 2,500 pg), or 3,500 to 4,500 pg of the chimeric Clostridial neurotoxin. Preferably, the unit dose contains 4,000 pg of the chimeric Clostridial neurotoxin.

[0275] A suitable unit dose may be 0.2 units up to 707 units of chimeric Clostridial neurotoxin. The upper end of the range may be 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, or 100 units of chimeric Clostridial neurotoxin, preferably 666 units. The lower end of the range may be 40, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, or 700 units of chimeric Clostridial neurotoxin, preferably 42 units or 31 units. The lower end of the range may be greater than 125 units. Preferably, the unit dose is 31 to 707 units of the chimeric Clostridial neurotoxin. The unit dose of the chimeric Clostridial neurotoxin may be 166 to 707 units. More preferably, the unit dose is 42 to 666 units of the chimeric Clostridial neurotoxin, for example, 200 to 400 units of the chimeric Clostridial neurotoxin, or 41 to 229 units, for example, 83 to 188 units, 83 to 125 units (e.g., 104 units), or 145 to 188 units of the chimeric Clostridial neurotoxin. Preferably, the unit dose is 166 units of the chimeric Clostridial neurotoxin. The unit dose can be 47 to 707 units of a chimeric Clostridial neurotoxin, e.g., 187 to 282 units of a chimeric Clostridial neurotoxin, or 47 to 258 units, e.g., 94 to 211 units, 94 to 141 units (e.g., 117 units), or 164 to 211 units of a chimeric Clostridial neurotoxin. The unit dose can be 188 units of a chimeric Clostridial neurotoxin.

[0276] A suitable unit dose may be between 1,000 pg and 5,500 pg. The upper end of the unit dose range may be 5,250, 5,200, 5,100, 5,000, 4,500, 4,000, 3,500, 3,000, 2,500, or 2,000 pg of chimeric Clostridial neurotoxin, preferably with an upper limit of 5,100 pg, and more preferably 5,000 pg. The lower limit of the unit dose range may be 1,100, 1,200, 1,250, 1,300, 1,350, 1,400, 1,450, 1,500, 2,000, 2,500, 3,000, 3,500, or 4,000 pg of chimeric Clostridial neurotoxin, preferably 1,400 pg, more preferably 1,500 pg. The lower limit of the range may be greater than 3,000 pg. The unit dose may be 1,400 pg to 5,100 pg (e.g., 1,500 pg to 5,000 pg), 2,000 pg to 5,100 pg, 3,000 pg to 5,100 pg, or 3,000 pg to 4,000 pg of chimeric Clostridial neurotoxin. The unit dose can contain more than 3,000 pg, up to 5,500 pg, of the chimeric Clostridial neurotoxin. The unit dose of the chimeric Clostridial neurotoxin can be 2,000 pg to 4,500 pg, 2,000 pg to 3,000 pg (e.g., 2,500 pg), or 3,500 to 4,500 pg of the chimeric Clostridial neurotoxin. Preferably, the unit dose contains 4,000 pg of the chimeric Clostridial neurotoxin.

[0277] Suitable unit doses may be between 42 and 258 units (e.g., up to 229 units). The upper end of the unit dose range may be 225, 220, 215, 210, 205, 200, 190, 180, 170, 160, 150, 125, 100, or 83 units of the chimeric clostridial neurotoxin, preferably with an upper limit of 212 units and more preferably 208 units. The lower end of the unit dose range may be 46, 50, 55, 60, 65, 70, 75, 80, 90, 100, 110, 120, 130, 140, 150, 160, or 166 units of the chimeric clostridial neurotoxin, preferably with a lower limit of 58 units and more preferably 62 units. The lower end of the range may be greater than 125 units. A unit dose may be 58 to 212 units (e.g., 62 to 208 units), 83 to 212 units, 125 to 212 units, or 125 to 166 units of a chimeric Clostridial neurotoxin. A unit dose may contain more than 125 units, up to 229 units of a chimeric Clostridial neurotoxin. A unit dose of a chimeric Clostridial neurotoxin may be 83 to 188 units, 83 to 125 units (e.g., 104 units), or 146 to 188 units of a chimeric Clostridial neurotoxin. Preferably, a unit dose contains 166 units of a chimeric Clostridial neurotoxin. A unit dose may contain 47 to 258 units of a chimeric Clostridial neurotoxin, for example, 94 to 211 units, 94 to 141 units (e.g., 117 units), or 164 to 211 units of a chimeric Clostridial neurotoxin. The unit dose may be 188 units of the chimeric clostridial neurotoxin.

[0278] The total dose administered per treatment session may be up to 255,000 pg of chimeric Clostridial neurotoxin, which may correspond to 15 times the unit dose. The total dose administered may be up to 255,000 pg of chimeric Clostridial neurotoxin, which may correspond to 28 times, 31 times, or 39 times the unit dose. In other words, the total amount of chimeric Clostridial neurotoxin administered in a given treatment session may be up to 255,000 pg. The total dose may be up to 240,000, 220,000, 200,000, 180,000, 160,000, 140,000, 110,000, 100,000, 90,000, 80,000, 70,000, 60,000, 50,000, 40,000, 30,000, 20,000, 10,000, or 5,000 pg. Preferably, the total dose may be up to 240,000 pg of chimeric Clostridial neurotoxin. The total dose may be at least 900, 1,000, 2,000, 3,000, 4,000, 5,000, 7,500, 10,000, 12,500, 15,000, 20,000, 30,000, 40,000, 50,000, 60,000, 70,000, 80,000, 90,000, 100,000, 120,000, 150,000, 175,000, 200,000, or 220,000 pg. Preferably, the total dose may be at least 1,500 pg, more preferably at least 2,000 pg of chimeric Clostridial neurotoxin, and more preferably greater than 3,000 pg, e.g., at least 12,000 pg. The total dose may be between 3,640 pg and 255,000 pg of chimeric Clostridial neurotoxin. The total dose may be between 2,000 and 240,000 pg, preferably between 128,000 and 240,000 pg. More preferably, the total dose administered is between 15,000 and 240,000 pg. The total dose may be 75,000 pg or 115,000 pg. The total dose may be 70,000 pg or 112,000 pg.

[0279] The total dose administered per treatment session may be up to 10,607 units of the chimeric Clostridial neurotoxin, which may correspond to 15 times the unit dose. The total dose administered may be up to 10,607 units of the chimeric Clostridial neurotoxin, which may correspond to 28 times, 31 times, or 39 times the unit dose. In other words, the total amount of the chimeric Clostridial neurotoxin administered in a given treatment session may be up to 10,607 units. The total dose may be up to 10,500, 10,000, 9,500, 9,000, 8,500, 8,000, 7,500, 7,000, 6,500, 6,000, 5,500, 5,000, 4,500, 4,000, 3,500, 3,000, 2,500, 2,000, 1,500, 1,000, 500, or 207 units. Preferably, the total dose may be up to 11,268 or 9,983 units of the chimeric Clostridial neurotoxin. The total dose may be at least 37, 50, 100, 150, 200, 250, 500, 1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, 5,000, 5,500, 6,000, 6,500, 7,000, 7,500, 8,000, 8,500, 9,000, 9,500, 9,151, 10,000, or 10,328 units. Preferably, the total dose is at least 62 or 70 units, more preferably at least 83 or 94 units of the chimeric Clostridial neurotoxin, and even more preferably greater than 125 or 141 units, e.g., at least 499 or 563 units. The total dose may be 165 Units to 10,607 Units, or 171 Units to 10,607 Units of the chimeric Clostridial neurotoxin. The total dose may be 83 to 9,983 Units or 94 to 10,607 Units, preferably 5,324 to 9,983 Units or 6,009 to 10,607 Units. More preferably, the total dose administered is 624 to 9,983 Units or 704 to 10,607 Units. The total dose may be 3,120 or 4,784 Units. The total dose may be 2,911 or 4,659 Units. The total dose may be 3,521 or 5,399 Units. The total dose may be 3,286 Units or 5,258 Units.

[0280] A suitable unit dose may be 2,500 pg, and the total dose may be up to 70,000 pg. For example, a suitable unit dose may be 2,500 pg, and the total dose may be 70,000 pg. A suitable unit dose may be 4,000 pg, and the total dose may be up to 112,000 pg. For example, a suitable unit dose may be 4,000 pg, and the total dose may be 112,000 pg. A suitable unit dose may be 5,000 pg, and the total dose may be up to 155,000 pg. For example, a suitable unit dose may be 5,000 pg, and the total dose may be 155,000 pg.

[0281] A suitable unit dose may be 104 units, and the total dose may be up to 2,912 units. For example, a suitable unit dose may be 104 units, and the total dose may be 2,912 units. A suitable unit dose may be 166 units, and the total dose may be up to 4,659 units. For example, a suitable unit dose may be 166 units, and the total dose may be 4,659 units. A suitable unit dose may be 208 units, and the total dose may be up to 6,448 units. For example, a suitable unit dose may be 208 units, and the total dose may be 6,448 units.

[0282] A suitable unit dose may be 117 units, and the total dose may be up to 3,286 units. For example, a suitable unit dose may be 117 units, and the total dose may be 3,286 units. A suitable unit dose may be 188 units, and the total dose may be up to 5,258 units. For example, a suitable unit dose may be 188 units, and the total dose may be 5,258 units. A suitable unit dose may be 235 units, and the total dose may be up to 7,277 units. For example, a suitable unit dose may be 235 units, and the total dose may be 7,277 units.

[0283] The total number of unit doses administered in a given treatment may be up to 15 times the number of unit doses. For example, the total number of unit doses administered may be up to 14 times, 13 times, 12 times, 11 times, 10 times, 9 times, 8 times, or 7 times the number of unit doses. The total number of unit doses administered may be at least 2 times, 3 times, 4 times, 5 times, 6 times, or 7 times the number of unit doses, and preferably at least 2 times the number of unit doses. The total number of unit doses administered may be 2 to 15 times, 7 to 15 times, or 10 to 14 times the number of unit doses. Preferably, the number of unit doses administered is 15 times the number of unit doses.

[0284] The total number of unit doses administered in a given treatment may be up to 39 times the unit dose (as long as the total dose administered during the treatment does not exceed the upper limit of 255,000 pg or 10,607 units). For example, the total number of unit doses administered may be up to 35, 31, 30, 29, 28, 27, 26, 25, or 20 times the unit dose, and preferably the total number of unit doses administered is up to 28 times the unit dose. The total number of unit doses administered may be at least 2, 3, 4, 5, 6, or 7 times the unit dose, and preferably at least 2 times. The total number of unit doses administered may be 2 to 39, 15 to 31, or 28 to 31. The total number of unit doses administered may be 28, 31, or 39.

[0285] Thus, the total dose administered per treatment session can be up to 192,500 pg of chimeric Clostridial neurotoxin. For example, the total dose administered can be up to 180,000 pg or up to 177,000 pg (e.g., up to 175,000 pg).

[0286] Thus, the total dose administered per treatment session can be up to 8,007 units of the chimeric Clostridial neurotoxin. For example, the total dose administered can be up to 7,488 units or up to 7,363 units (e.g., up to 7,280 units). The total dose administered per treatment session can be up to 9,037 units of the chimeric Clostridial neurotoxin. For example, the total dose administered can be up to 8,451 units or up to 8,310 units (e.g., up to 8,216 units).

[0287] The total dose administered per treatment session can be up to 110,000 pg of chimeric Clostridial neurotoxin. For example, the total dose administered can be up to 105,000 pg, up to 102,000 pg (e.g., up to 100,000 pg).

[0288] The total dose administered per treatment session may be up to 4,576 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 4,368 units, preferably up to 4,243 units (more preferably up to 4,160 units). The total dose administered per treatment session may be up to 5,165 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 4,929 units, preferably up to 4,789 units (more preferably up to 4,695 units).

[0289] The term "up to," when used in reference to a value (e.g., up to 255,000 pg), means up to and including the stated value. Thus, by way of example, a reference to administering "up to 255,000 pg" of a chimeric Clostridial neurotoxin encompasses the administration of 255,000 pg of a chimeric Clostridial neurotoxin, as well as the administration of less than 255,000 pg of a chimeric Clostridial neurotoxin.

[0290] When the disorder is headache (e.g., migraine pain) or migraine, at least a unit dose of the chimeric Clostridial neurotoxin may be administered to one or more of the frontalis muscle, corrugator muscle, nasal muscle, orbicularis oculi muscle, temporalis muscle, occipital muscle, and trapezius muscle. Preferably, at least a unit dose of the chimeric Clostridial neurotoxin may be administered to the frontalis muscle, corrugator muscle, nasal muscle, orbicularis oculi muscle, temporalis muscle, occipital muscle, and trapezius muscle. In some embodiments, multiple unit doses are administered to one or more of the frontalis muscle, corrugator muscle, nasal muscle, orbicularis oculi muscle, temporalis muscle, occipital muscle, and trapezius muscle. In one embodiment, a single unit dose is administered to the corrugator muscle, nasal muscle, and orbicularis oculi muscle, and multiple unit doses are administered to the frontalis muscle, temporalis muscle, occipital muscle, and trapezius muscle. The multiple unit doses may be 2 to 10 unit doses, for example 2 to 8 unit doses, preferably 2 to 5 unit doses, for example 2 to 4 unit doses.

[0291] More preferably, the method comprises administering a chimeric Clostridial neurotoxin to at least one of the frontalis, corrugator (e.g., corrugator supercilii), nasal, orbicularis oculi, temporalis, occipitalis, or trapezius muscles. The method may comprise administering a chimeric Clostridial neurotoxin to the frontalis, corrugator (e.g., corrugator supercilii), nasal, orbicularis oculi, temporalis, occipitalis, and trapezius muscles.

[0292] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 2 unit doses into the frontalis muscle (preferably 2 unit doses per frontalis muscle); (ii) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iii) one unit dose to the bridge of the nose (preferably one unit dose per bridge of the nose); (iv) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (v) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (vi) 3 unit doses into the occipitalis muscle (preferably 3 unit doses per occipitalis muscle); and / or (vii) 2 unit doses into the trapezius muscle (preferably 2 unit doses per trapezius muscle).

[0293] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 2 unit doses into the frontalis muscle (preferably 2 unit doses per frontalis muscle); (ii) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iii) one unit dose to the bridge of the nose (preferably one unit dose per bridge of the nose); (iv) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (v) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (vi) 3 unit doses into the occipitalis muscle (preferably 3 unit doses per occipitalis muscle); and (vii) 2 unit doses into the trapezius muscle (preferably 2 unit doses per trapezius muscle).

[0294] Treatment of headache (e.g., migraine pain) or migraine headache can include administering a unit dose of a chimeric Clostridial neurotoxin bilaterally. Treatment of headache (e.g., migraine pain) or migraine headache can include administering: (i) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (ii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iii) two unit doses to the bridge of the nose (preferably one unit dose to the bridge of the nose on a first side of the face and one unit dose to the bridge of the nose on a second side of the face); (iv) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (v) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (vi) 6 unit doses to the occipitalis muscle (preferably 3 unit doses to the occipitalis muscle on the first side of the head and 3 unit doses to the occipitalis muscle on the second side of the head); and / or (vii) 4 unit doses to the trapezius muscle (preferably 2 unit doses to the trapezius muscle on a first side of the neck and 2 unit doses to the trapezius muscle on a second side of the neck).

[0295] Preferably, the treatment of headache (e.g., migraine pain) or migraine headache comprises the administration of: (i) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (ii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iii) two unit doses to the bridge of the nose (preferably one unit dose to the bridge of the nose on a first side of the face and one unit dose to the bridge of the nose on a second side of the face); (iv) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (v) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (vi) 6 unit doses to the occipitalis muscle (preferably 3 unit doses to the occipitalis muscle on the first side of the head and 3 unit doses to the occipitalis muscle on the second side of the head); and (vii) 4 unit doses to the trapezius muscle (preferably 2 unit doses to the trapezius muscle on a first side of the neck and 2 unit doses to the trapezius muscle on a second side of the neck).

[0296] When treating headache (e.g., migraine pain) or migraine headache as described in the above embodiments, it is preferred to administer one unit dose per injection (e.g., injection site). Thus, administration of a chimeric Clostridial neurotoxin can include: (i) two injections into the frontalis muscle (preferably two injections per frontalis muscle); (ii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iii) a single injection into the bridge of the nose (preferably one injection per bridge of the nose); (iv) one injection into the orbicularis oculi muscle (preferably one injection per orbicularis oculi muscle); (v) four injections into the temporalis muscle (preferably four injections per temporalis muscle); (vi) three injections into the occipitalis muscle (preferably three injections per occipitalis muscle); and / or (vii) Two injections into the trapezius muscle (preferably two injections per trapezius muscle).

[0297] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably two injections per frontalis muscle); (ii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iii) a single injection into the bridge of the nose (preferably one injection per bridge of the nose); (iv) one injection into the orbicularis oculi muscle (preferably one injection per orbicularis oculi muscle); (v) four injections into the temporalis muscle (preferably four injections per temporalis muscle); (vi) three injections into the occipitalis muscle (preferably, three injections per occipitalis muscle); and (vii) Two injections into the trapezius muscle (preferably two injections per trapezius muscle).

[0298] Administration of the chimeric Clostridial neurotoxin may include: (i) four injections into the frontalis muscle (preferably two injections into the frontalis muscle on the first side of the face and two injections into the frontalis muscle on the second side of the face); (ii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iii) two injections into the nasal bridge (preferably one injection into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face; (iv) two injections into the orbicularis oculi muscle (preferably one injection into the orbicularis oculi muscle on a first side of the face and one injection into the orbicularis oculi muscle on a second side of the face); (v) eight injections into the temporalis muscle (preferably four injections into the temporalis muscle on the first side of the head and four injections into the temporalis muscle on the second side of the head); (vi) six injections into the occipitalis muscle (preferably three injections into the occipitalis muscle on the first side of the head and three injections into the occipitalis muscle on the second side of the head); and / or (vii) Four injections into the trapezius muscle (preferably two injections into the trapezius muscle on the first side of the neck and two injections into the trapezius muscle on the second side of the neck).

[0299] Preferably, the administration of the chimeric Clostridial neurotoxin comprises: (i) four injections into the frontalis muscle (preferably two injections into the frontalis muscle on the first side of the face and two injections into the frontalis muscle on the second side of the face); (ii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iii) two injections into the nasal bridge (preferably one injection into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face; (iv) two injections into the orbicularis oculi muscle (preferably one injection into the orbicularis oculi muscle on a first side of the face and one injection into the orbicularis oculi muscle on a second side of the face); (v) eight injections into the temporalis muscle (preferably four injections into the temporalis muscle on the first side of the head and four injections into the temporalis muscle on the second side of the head); (vi) six injections into the occipitalis muscle (preferably three injections into the occipitalis muscle on the first side of the head and three injections into the occipitalis muscle on the second side of the head); and (vii) Four injections into the trapezius muscle (preferably two injections into the trapezius muscle on the first side of the neck and two injections into the trapezius muscle on the second side of the neck).

[0300] When the disorder is headache (e.g., migraine pain) or migraine, at least a unit dose of the chimeric Clostridial neurotoxin can be administered intramuscularly or intradermally to one or more of the frontalis, procerus, corrugator, temporalis, occipitalis, trapezius, and cervical paraspinal muscles. Preferably, at least a unit dose of the chimeric Clostridial neurotoxin can be administered intramuscularly or intradermally to the frontalis, procerus, corrugator, temporalis, occipitalis, trapezius, and cervical paraspinal muscles (e.g., at least a unit dose to each of the cervical paraspinal muscles).

[0301] When the disorder is headache (e.g., migraine pain) or migraine, at least a unit dose of the chimeric Clostridial neurotoxin can be administered to one or more of the frontalis, procerus, corrugator, temporalis, occipitalis, trapezius, and cervical paraspinal muscles. Preferably, at least a unit dose of the chimeric Clostridial neurotoxin can be administered to the frontalis, procerus, corrugator, temporalis, occipitalis, trapezius, and cervical paraspinal muscles (e.g., at least a unit dose to each of the cervical paraspinal muscles). In one embodiment: (i) a single unit dose is administered to one or more of the procerus and corrugator muscles (preferably, a single unit dose is administered to the corrugator muscles on a first side of the face (e.g., the left side) and a second unit dose is administered to the corrugator muscles on a second side of the face (e.g., the right side)); and / or (preferably, and) (iii) Multiple unit doses are administered to one or more of the frontalis, temporalis, occipitalis, trapezius, and cervical paraspinal muscles (e.g., where one or two unit doses are administered to each muscle of the cervical paraspinal muscles). The multiple unit doses can be 2 to 8 unit doses, e.g., 2 to 5 unit doses.

[0302] Treatment of headache (e.g., migraine pain) or migraine headache may include administering a unit dose of a chimeric Clostridial neurotoxin bilaterally intramuscularly or intradermally. Treatment of headache (e.g., migraine pain) or migraine headache may include administering: (i) 2 unit doses to the frontalis muscle on the first side of the face and / or 2 unit doses to the frontalis muscle on the second side of the face; (ii) 1 unit dose into the procerus muscle; (iii) one unit dose to the corrugator muscles on a first side of the face and / or one unit dose to the corrugator muscles on a second side of the face; (iv) 4 unit doses to the temporalis muscle on the first side of the head and / or 4 unit doses to the temporalis muscle on the second side of the head; (v) 3 unit doses to the occipitalis muscle on a first side of the neck / head (preferably the head) and / or 3 unit doses to the occipitalis muscle on a second side of the neck / head (preferably the head); (vi) 3 unit doses to the trapezius muscle on a first side of the neck and / or 3 unit doses to the trapezius muscle on a second side of the neck; and / or (vii) 4 unit doses to the cervical paraspinal group on a first side of the neck and / or 4 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 1 unit dose is administered per cervical paraspinal muscle group), or 2 unit doses to the cervical paraspinal group on a first side of the neck and / or 2 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 2 unit doses are administered per cervical paraspinal muscle group).

[0303] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 2 unit doses to the frontalis muscle on the first side of the face and 2 unit doses to the frontalis muscle on the second side of the face; (ii) 1 unit dose into the procerus muscle; (iii) 1 unit dose to a first lateral facial corrugator muscle and 1 unit dose to a second lateral facial corrugator muscle; (iv) a 4-unit dose to the temporalis muscle on the first side of the head and a 4-unit dose to the temporalis muscle on the second side of the head; (v) 3 unit doses to the occipitalis muscle on a first side of the neck / head (preferably the head) and 3 unit doses to the occipitalis muscle on a second side of the neck / head (preferably the head); (vi) 3 unit doses to the trapezius muscle on a first side of the neck and 3 unit doses to the trapezius muscle on a second side of the neck; and / or (vii) 4 unit doses to the cervical paraspinal group on a first side of the neck and 4 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 1 unit dose is administered per cervical paraspinal muscle group), or 2 unit doses to the cervical paraspinal group on a first side of the neck and 2 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 2 unit doses are administered per cervical paraspinal muscle group).

[0304] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 2 unit doses to the frontalis muscle on the first side of the face and 2 unit doses to the frontalis muscle on the second side of the face; (ii) 1 unit dose into the procerus muscle; (iii) 1 unit dose to a first lateral facial corrugator muscle and 1 unit dose to a second lateral facial corrugator muscle; (iv) a 4-unit dose to the temporalis muscle on the first side of the head and a 4-unit dose to the temporalis muscle on the second side of the head; (v) 3 unit doses to the occipitalis muscle on a first side of the neck / head (preferably the head) and 3 unit doses to the occipitalis muscle on a second side of the neck / head (preferably the head); (vi) 3 unit doses to the trapezius muscle on a first side of the neck and 3 unit doses to the trapezius muscle on a second side of the neck; and (vii) 4 unit doses to the cervical paraspinal group on a first side of the neck and 4 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 1 unit dose is administered per cervical paraspinal muscle group), or 2 unit doses to the cervical paraspinal group on a first side of the neck and 2 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 2 unit doses are administered per cervical paraspinal muscle group).

[0305] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 1 unit dose into the frontalis muscle (preferably 1 unit dose per frontalis muscle); (ii) one unit dose into the procerus (preferably one unit dose per procerus); (iii) 1 unit dose to the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iv) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (v) 3 unit doses into the occipitalis muscle (preferably 3 unit doses per occipitalis muscle); (v) 3 unit doses into the trapezius muscle (preferably 3 unit doses per trapezius muscle); and / or (vii) 4 unit doses in the cervical paraspinal group (preferably 4 unit doses per cervical paraspinal group), or 2 unit doses in the cervical paraspinal group (preferably 2 unit doses per cervical paraspinal group).

[0306] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 1 unit dose into the frontalis muscle (preferably 1 unit dose per frontalis muscle); (ii) one unit dose into the procerus (preferably one unit dose per procerus); (iii) 1 unit dose to the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iv) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (v) 3 unit doses into the occipitalis muscle (preferably 3 unit doses per occipitalis muscle); (vi) 3 unit doses into the trapezius muscle (preferably 3 unit doses per trapezius muscle); and (vii) 4 unit doses in the cervical paraspinal group (preferably 4 unit doses per cervical paraspinal group), or 2 unit doses in the cervical paraspinal group (preferably 2 unit doses per cervical paraspinal group).

[0307] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (ii) 1 unit dose into the procerus muscle; (iii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iv) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (v) 6 unit doses to the occipitalis muscle (preferably 3 unit doses to the occipitalis muscle on the first side of the head and 3 unit doses to the occipitalis muscle on the second side of the head); (vi) 6 unit doses to the trapezius muscle (preferably 3 unit doses to the trapezius muscle on a first side of the neck and 3 unit doses to the trapezius muscle on a second side of the neck); and / or (vii) 8 unit doses to the cervical paraspinal group (preferably 4 unit doses to the cervical paraspinal group on a first side of the neck and 4 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 1 unit dose is administered per cervical paraspinal muscle group)), or 4 unit doses to the cervical paraspinal group (preferably 2 unit doses to the cervical paraspinal group on a first side of the neck and 2 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 2 unit doses are administered per cervical paraspinal muscle group).

[0308] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (ii) 1 unit dose into the procerus muscle; (iii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iv) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (v) 6 unit doses to the occipitalis muscle (preferably 3 unit doses to the occipitalis muscle on the first side of the head and 3 unit doses to the occipitalis muscle on the second side of the head); (vi) 6 unit doses to the trapezius muscle (preferably 3 unit doses to the trapezius muscle on a first side of the neck and 3 unit doses to the trapezius muscle on a second side of the neck); and (vii) 8 unit doses to the cervical paraspinal group (preferably 4 unit doses to the cervical paraspinal group on a first side of the neck and 4 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 1 unit dose is administered per cervical paraspinal muscle group)), or 4 unit doses to the cervical paraspinal group (preferably 2 unit doses to the cervical paraspinal group on a first side of the neck and 2 unit doses to the cervical paraspinal group on a second side of the neck (e.g., when 2 unit doses are administered per cervical paraspinal muscle group).

[0309] When treating headache (e.g., migraine pain) or migraine headache as described in the above embodiments, it is preferred to administer one unit dose per injection site. Thus, treatment can include administering a chimeric Clostridial neurotoxin to: (i) two injection sites in the frontalis muscle on a first side of the face and / or two injection sites in the frontalis muscle on a second side of the face; (ii) one injection site in the procerus muscle; (iii) one injection site in the corrugator muscle of a first side of the face and / or one injection site in the corrugator muscle of a second side of the face; (iv) four injection sites in the temporalis muscle on the first side of the head and / or four injection sites in the temporalis muscle on the second side of the head; (v) three injection sites in the occipitalis muscle on a first side of the neck / head (preferably the head) and / or three injection sites in the occipitalis muscle on a second side of the neck / head (preferably the head); (vi) three injection sites in the trapezius muscle on a first side of the neck and / or three injection sites in the trapezius muscle on a second side of the neck; and / or (vii) Four injection sites in the cervical paraspinal group on a first side of the neck and / or four injection sites in the cervical paraspinal group on a second side of the neck (e.g., where there is one injection site per cervical paraspinal muscle group), or two injection sites in the cervical paraspinal group on a first side of the neck and / or two injection sites in the cervical paraspinal group on a second side of the neck (e.g., where there is two injection sites per cervical paraspinal muscle group).

[0310] Treatment can include administering a chimeric Clostridial neurotoxin to: (i) two injection sites in the frontalis muscle on the first side of the face and two injection sites in the frontalis muscle on the second side of the face; (ii) one injection site in the procerus muscle; (iii) one injection site in the first lateral facial corrugator muscle and one injection site in the second lateral facial corrugator muscle; (iv) four injection sites in the temporalis muscle on the first side of the head and four injection sites in the temporalis muscle on the second side of the head; (v) three injection sites in the occipitalis muscle on a first side of the neck / head (preferably the head) and three injection sites in the occipitalis muscle on a second side of the neck / head (preferably the head); (vi) three injection sites in the trapezius muscle on a first side of the neck and three injection sites in the trapezius muscle on a second side of the neck; and / or (vii) Four injection sites in the cervical paraspinal group on a first side of the neck and four injection sites in the cervical paraspinal group on a second side of the neck (e.g., one injection site per cervical paraspinal muscle group), or two injection sites in the cervical paraspinal group on a first side of the neck and two injection sites in the cervical paraspinal group on a second side of the neck (e.g., two injection sites per cervical paraspinal muscle group).

[0311] Treatment can include administering a chimeric Clostridial neurotoxin to: (i) two injection sites in the frontalis muscle on the first side of the face and two injection sites in the frontalis muscle on the second side of the face; (ii) one injection site in the procerus muscle; (iii) one injection site in the first lateral facial corrugator muscle and one injection site in the second lateral facial corrugator muscle; (iv) four injection sites in the temporalis muscle on the first side of the head and four injection sites in the temporalis muscle on the second side of the head; (v) three injection sites in the occipitalis muscle on a first side of the neck / head (preferably the head) and three injection sites in the occipitalis muscle on a second side of the neck / head (preferably the head); (vi) three injection sites in the trapezius muscle on a first side of the neck and three injection sites in the trapezius muscle on a second side of the neck; and (vii) Four injection sites in the cervical paraspinal group on a first side of the neck and four injection sites in the cervical paraspinal group on a second side of the neck (e.g., one injection site per cervical paraspinal muscle group), or two injection sites in the cervical paraspinal group on a first side of the neck and two injection sites in the cervical paraspinal group on a second side of the neck (e.g., two injection sites per cervical paraspinal muscle group).

[0312] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably two injections per frontalis muscle); (ii) a single injection into the procerus muscle (preferably one injection per procerus muscle); (iii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iv) four injections into the temporalis muscle (preferably four injections per temporalis muscle); (v) three injections into the occipitalis muscle (preferably three injections per occipitalis muscle); (vi) three injections into the trapezius muscle (preferably three injections per trapezius muscle); and / or (vii) Four injections into the cervical paraspinal group (preferably two injections per cervical paraspinal group).

[0313] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably two injections per frontalis muscle); (ii) a single injection into the procerus muscle (preferably one injection per procerus muscle); (iii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iv) four injections into the temporalis muscle (preferably four injections per temporalis muscle); (v) three injections into the occipitalis muscle (preferably three injections per occipitalis muscle); (vi) three injections into the trapezius muscle (preferably, three injections per trapezius muscle); and (vii) Four injections into the cervical paraspinal group (preferably two injections per cervical paraspinal group).

[0314] Administration of the chimeric Clostridial neurotoxin may include: (i) four injections into the frontalis muscle (preferably two injections into the frontalis muscle on the first side of the face and two injections into the frontalis muscle on the second side of the face); (ii) a single injection into the procerus muscle; (iii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iv) eight injections into the temporalis muscle (preferably four injections into the temporalis muscle on the first side of the head and four injections into the temporalis muscle on the second side of the head); (v) six injections into the occipitalis muscle (preferably three injections into the occipitalis muscle on the first side of the head and three injections into the occipitalis muscle on the second side of the head); (vi) six injections into the trapezius muscle (preferably three injections into the trapezius muscle on the first side of the neck and three injections into the trapezius muscle on the second side of the neck); and / or (vii) Eight injections into the cervical paraspinal groups (preferably four injections into the cervical paraspinal groups on the first side of the neck and four injections into the cervical paraspinal groups on the second side of the neck (e.g., one injection per cervical paraspinal muscle group) or four injections into the cervical paraspinal groups (preferably two injections into the cervical paraspinal groups on the first side of the neck and two injections into the cervical paraspinal groups on the second side of the neck (e.g., two injections per cervical paraspinal muscle group)).

[0315] Administration of the chimeric Clostridial neurotoxin may include: (i) four injections into the frontalis muscle (preferably two injections into the frontalis muscle on the first side of the face and two injections into the frontalis muscle on the second side of the face); (ii) a single injection into the procerus muscle; (iii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iv) eight injections into the temporalis muscle (preferably four injections into the temporalis muscle on the first side of the head and four injections into the temporalis muscle on the second side of the head); (v) six injections into the occipitalis muscle (preferably three injections into the occipitalis muscle on the first side of the head and three injections into the occipitalis muscle on the second side of the head); (vi) six injections into the trapezius muscle (preferably three injections into the trapezius muscle on the first side of the neck and three injections into the trapezius muscle on the second side of the neck); and (vii) Eight injections into the cervical paraspinal groups (preferably four injections into the cervical paraspinal groups on the first side of the neck and four injections into the cervical paraspinal groups on the second side of the neck (e.g., if there is one injection site per cervical paraspinal muscle group) or four injections into the cervical paraspinal groups (preferably two injections into the cervical paraspinal groups on the first side of the neck and two injections into the cervical paraspinal groups on the second side of the neck (e.g., if there are two injections per cervical paraspinal muscle group)).

[0316] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 2 unit doses into the frontalis muscle (preferably 2 unit doses per frontalis muscle); (ii) 1 unit dose into the procerus muscle; (iii) 1 unit dose to the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iv) 5 unit doses into the temporalis muscle (preferably 5 unit doses per temporalis muscle); (v) 4 unit doses into the occipitalis muscle (preferably 4 unit doses per occipitalis muscle); (vi) 5 unit doses into the trapezius muscle (preferably 5 unit doses per trapezius muscle); and / or (vii) 4 unit dose sites in the cervical paraspinal group (preferably 4 unit doses per cervical paraspinal group), or 2 unit dose sites in the cervical paraspinal group (preferably 2 unit doses per cervical paraspinal group).

[0317] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 2 unit doses into the frontalis muscle (preferably 2 unit doses per frontalis muscle); (ii) one unit dose into the procerus (preferably one unit dose per procerus); (iii) 1 unit dose to the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iv) 5 unit doses into the temporalis muscle (preferably 5 unit doses per temporalis muscle); (v) 4 unit doses into the occipitalis muscle (preferably 4 unit doses per occipitalis muscle); (vi) 5 unit doses into the trapezius muscle (preferably 5 unit doses per trapezius muscle); and (vii) 4 unit doses in the cervical paraspinal group (preferably 4 unit doses per cervical paraspinal group), or 2 unit dose sites in the cervical paraspinal group (preferably 2 unit doses per cervical paraspinal group).

[0318] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (ii) 1 unit dose into the procerus muscle; (iii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iv) 10 unit doses to the temporalis muscle (preferably 5 unit doses to the temporalis muscle on the first side of the head and 5 unit doses to the temporalis muscle on the second side of the head); (v) 8 unit doses to the occipitalis muscle (preferably 4 unit doses to the occipitalis muscle on the first side of the head and 4 unit doses to the occipitalis muscle on the second side of the head); (vi) 10 unit doses to the trapezius muscle (preferably 4 unit doses to the trapezius muscle on a first side of the neck and 4 unit doses to the trapezius muscle on a second side of the neck); and / or (vii) 4 unit doses to the cervical paraspinal muscles (e.g., 1 or 2 unit doses administered per cervical paraspinal muscle group).

[0319] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (ii) 1 unit dose into the procerus muscle; (iii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iv) 10 unit doses to the temporalis muscle (preferably 5 unit doses to the temporalis muscle on the first side of the head and 5 unit doses to the temporalis muscle on the second side of the head); (v) 8 unit doses to the occipitalis muscle (preferably 4 unit doses to the occipitalis muscle on the first side of the head and 4 unit doses to the occipitalis muscle on the second side of the head); (vi) 10 unit doses to the trapezius muscle (preferably 5 unit doses to the trapezius muscle on a first side of the neck and 5 unit doses to the trapezius muscle on a second side of the neck); and (vii) 4 unit doses to the cervical paraspinal muscles (e.g., 1 or 2 unit doses administered per cervical paraspinal muscle group).

[0320] When treating headache (e.g., migraine pain) or migraine headache, administration of a chimeric Clostridial neurotoxin can include: (i) two injections into the frontalis muscle (preferably two injections per frontalis muscle); (ii) a single injection into the procerus muscle; (iii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iv) five injections into the temporalis muscle (preferably five injections per temporalis muscle); (v) four injections into the occipitalis muscle (preferably four injections per occipitalis muscle); (vi) 5 injections into the trapezius muscle (preferably 5 injections per trapezius muscle); and / or (vii) Four injections into the cervical paraspinal group (e.g., one or two injections administered per cervical paraspinal muscle group).

[0321] Administration may include: (i) two injections into the frontalis muscle (preferably two injections per frontalis muscle); (ii) a single injection into the procerus muscle; (iii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iv) five injections into the temporalis muscle (preferably five injections per temporalis muscle); (v) four injections into the occipitalis muscle (preferably four injections per occipitalis muscle); (vi) 5 injections into the trapezius muscle (preferably 5 injections per trapezius muscle); and (vii) Four injections into the cervical paraspinal group (e.g., one or two injections administered per cervical paraspinal muscle group).

[0322] Administration may include: (i) four injections into the frontalis muscle (preferably two injections into the frontalis muscle on the first side of the face and two injections into the frontalis muscle on the second side of the face); (ii) a single injection into the procerus muscle; (iii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iv) 10 injections into the temporalis muscle (preferably 5 injections into the temporalis muscle on the first side of the head and 5 injections into the temporalis muscle on the second side of the head); (v) eight injections into the occipitalis muscle (preferably four injections into the occipitalis muscle on the first side of the head and four injections into the occipitalis muscle on the second side of the head); (vi) 10 injections into the trapezius muscle (preferably 4 injections into the trapezius muscle on the first side of the neck and 4 injections into the trapezius muscle on the second side of the neck); and / or (vii) Four injections into the cervical paraspinal group (e.g., one or two injections administered per cervical paraspinal muscle group).

[0323] Administration may include: (i) four injections into the frontalis muscle (preferably two injections into the frontalis muscle on the first side of the face and two injections into the frontalis muscle on the second side of the face); (ii) a single injection into the procerus muscle; (iii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iv) 10 injections into the temporalis muscle (preferably 5 injections into the temporalis muscle on the first side of the head and 5 injections into the temporalis muscle on the second side of the head); (v) eight injections into the occipitalis muscle (preferably four injections into the occipitalis muscle on the first side of the head and four injections into the occipitalis muscle on the second side of the head); (vi) 10 injections into the trapezius muscle (preferably 4 injections into the trapezius muscle on the first side of the neck and 4 injections into the trapezius muscle on the second side of the neck); (vii) Four injections into the cervical paraspinal group (e.g., one or two injections administered per cervical paraspinal muscle group).

[0324] If the disorder is headache (e.g., migraine pain) or migraine, at least a unit dose of the chimeric Clostridial neurotoxin may be administered to one or more of the frontalis, corrugator, nasal, orbicularis, masseter, temporalis, occipital, and trapezius muscles. In one embodiment, at least a unit dose of the chimeric Clostridial neurotoxin may be administered to the frontalis, corrugator, nasal, orbicularis, masseter, temporalis, occipital, and trapezius muscles. In one embodiment: (i) a single unit dose is administered to one or more of the frontalis muscle, corrugator muscles (preferably, a single unit dose is administered to the corrugator muscles on a first side of the face (e.g., the left side) and a second unit dose is administered to the corrugator muscles on a second side of the face (e.g., the right side)), orbicularis oculi muscle, masseter muscle, and / or (preferably, and) upper trapezius muscle); (ii) half of the unit dose is administered to the bridge of the nose; and / or (preferably, and) (iii) A plurality of unit doses is administered to one or more of the temporalis, occipitalis, and / or (preferably) lower trapezius muscles. The plurality of unit doses may be 2 to 6 unit doses, for example, 2 to 5 unit doses.

[0325] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 1 unit dose into the frontalis muscle (preferably 1 unit dose per frontalis muscle); (ii) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iii) 0.5 unit dose to the bridge of the nose (preferably 0.5 unit dose per bridge of the nose); (iv) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (v) 1 unit dose into the masseter muscles (preferably 1 unit dose per masseter muscle); (vi) 6 unit doses into the temporalis muscle (preferably 6 unit doses per temporalis muscle); (vii) 6 unit doses into the occipitalis muscle (preferably 6 unit doses per occipitalis muscle); (viii) 1 unit dose into the upper trapezius muscle (preferably 1 unit dose per upper trapezius muscle); and / or (ix) 2 unit doses into the lower trapezius muscle (preferably 2 unit doses per lower trapezius muscle).

[0326] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 1 unit dose into the frontalis muscle (preferably 1 unit dose per frontalis muscle); (ii) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iii) 0.5 unit dose to the bridge of the nose (preferably 0.5 unit dose per bridge of the nose); (iv) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (v) 1 unit dose into the masseter muscles (preferably 1 unit dose per masseter muscle); (vi) 6 unit doses into the temporalis muscle (preferably 6 unit doses per temporalis muscle); (vii) 6 unit doses into the occipitalis muscle (preferably 6 unit doses per occipitalis muscle); (viii) 1 unit dose into the upper trapezius muscle (preferably 1 unit dose per upper trapezius muscle); and (ix) 2 unit doses into the lower trapezius muscle (preferably 2 unit doses per lower trapezius muscle).

[0327] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) two unit doses to the frontalis muscle (preferably one unit dose to the frontalis muscle on a first side of the face and one unit dose to the frontalis muscle on a second side of the face); (ii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iii) 1 unit dose to the bridge of the nose (preferably 0.5 unit dose to the bridge of the nose on a first side of the face and 0.5 unit dose to the bridge of the nose on a second side of the face); (iv) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (v) two unit doses to the masseter muscles (preferably one unit dose to the masseter muscles on a first side of the face and one unit dose to the masseter muscles on a second side of the face); (vi) 12 unit doses to the temporalis muscle (preferably 6 unit doses to the temporalis muscle on the first side of the head and 6 unit doses to the temporalis muscle on the second side of the head); (vii) 12 unit doses to the occipitalis muscle (preferably 6 unit doses to the occipitalis muscle on a first side of the head and 6 unit doses to the occipitalis muscle on a second side of the head); (viii) two unit doses to the upper trapezius muscle (preferably one unit dose to the upper trapezius muscle on a first side of the neck and one unit dose to the upper trapezius muscle on a second side of the neck); and / or (ix) 4 unit doses to the lower trapezius muscle (preferably 2 unit doses to the lower trapezius muscle on a first neck side and / or 2 unit doses to the lower trapezius muscle on a second neck side).

[0328] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) two unit doses to the frontalis muscle (preferably one unit dose to the frontalis muscle on a first side of the face and one unit dose to the frontalis muscle on a second side of the face); (ii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iii) 1 unit dose to the bridge of the nose (preferably 0.5 unit dose to the bridge of the nose on a first side of the face and 0.5 unit dose to the bridge of the nose on a second side of the face); (iv) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (v) two unit doses to the masseter muscles (preferably one unit dose to the masseter muscles on a first side of the face and one unit dose to the masseter muscles on a second side of the face); (vi) 12 unit doses to the temporalis muscle (preferably 6 unit doses to the temporalis muscle on the first side of the head and 6 unit doses to the temporalis muscle on the second side of the head); (vii) 12 unit doses to the occipitalis muscle (preferably 6 unit doses to the occipitalis muscle on a first side of the head and 6 unit doses to the occipitalis muscle on a second side of the head); (viii) two unit doses to the upper trapezius muscle (preferably one unit dose to the upper trapezius muscle on a first side of the neck and one unit dose to the upper trapezius muscle on a second side of the neck); and (ix) 4 unit doses to the lower trapezius muscle (preferably 2 unit doses to the lower trapezius muscle on a first neck side and / or 2 unit doses to the lower trapezius muscle on a second neck side).

[0329] Administration of the chimeric Clostridial neurotoxin may include: (i) one injection into the frontalis muscle (preferably one injection per frontalis muscle); (ii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iii) a single injection into the bridge of the nose (preferably one injection per bridge of the nose); (iv) one injection into the orbicularis oculi muscle (preferably one injection per orbicularis oculi muscle); (v) one injection into the masseter muscles (preferably one injection per masseter muscle); (vi) three injections into the temporalis muscle (preferably three injections per temporalis muscle); (vii) three injections into the occipitalis muscle (preferably three injections per occipitalis muscle); (viii) one injection into the upper trapezius muscle (preferably one injection per upper trapezius muscle); and / or (ix) One injection into the lower trapezius muscle (preferably one injection per lower trapezius muscle).

[0330] Administration of the chimeric Clostridial neurotoxin may include: (i) one injection into the frontalis muscle (preferably one injection per frontalis muscle); (ii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iii) a single injection into the bridge of the nose (preferably one injection per bridge of the nose); (iv) one injection into the orbicularis oculi muscle (preferably one injection per orbicularis oculi muscle); (v) one injection into the masseter muscles (preferably one injection per masseter muscle); (vi) three injections into the temporalis muscle (preferably three injections per temporalis muscle); (vii) three injections into the occipitalis muscle (preferably three injections per occipitalis muscle); (viii) one injection into the upper trapezius muscle (preferably one injection per upper trapezius muscle); and (ix) One injection into the lower trapezius muscle (preferably one injection per lower trapezius muscle).

[0331] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably one injection into the frontalis muscle on a first side of the face and one injection into the frontalis muscle on a second side of the face); (ii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iii) two injections into the nasal bridge (preferably one injection into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face); (iv) two injections into the orbicularis oculi muscle (preferably one injection into the orbicularis oculi muscle on a first side of the face and one injection into the orbicularis oculi muscle on a second side of the face); (v) two injections into the masseter muscles (preferably one injection into the masseter muscles on a first side of the face and one injection into the masseter muscles on a second side of the face); (vi) six injections into the temporalis muscle (preferably three injections into the temporalis muscle on the first side of the head and three injections into the temporalis muscle on the second side of the head); (vii) six injections into the occipitalis muscle (preferably three injections into the occipitalis muscle on the first side of the head and three injections into the occipitalis muscle on the second side of the head); (viii) two injections into the upper trapezius muscle (preferably one injection into the upper trapezius muscle on a first side of the neck and one injection into the upper trapezius muscle on a second side of the neck); and / or (ix) Two injections into the lower trapezius muscle (preferably one injection into the lower trapezius muscle on a first side of the neck and / or one injection into the lower trapezius muscle on a second side of the neck).

[0332] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably one injection into the frontalis muscle on a first side of the face and one injection into the frontalis muscle on a second side of the face); (ii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iii) two injections into the nasal bridge (preferably one injection into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face); (iv) two injections into the orbicularis oculi muscle (preferably one injection into the orbicularis oculi muscle on a first side of the face and one injection into the orbicularis oculi muscle on a second side of the face); (v) two injections into the masseter muscles (preferably one injection into the masseter muscles on a first side of the face and one injection into the masseter muscles on a second side of the face); (vi) six injections into the temporalis muscle (preferably three injections into the temporalis muscle on the first side of the head and three injections into the temporalis muscle on the second side of the head); (vii) six injections into the occipitalis muscle (preferably three injections into the occipitalis muscle on the first side of the head and three injections into the occipitalis muscle on the second side of the head); (viii) two injections into the upper trapezius muscle (preferably one injection into the upper trapezius muscle on a first side of the neck and one injection into the upper trapezius muscle on a second side of the neck); and (ix) Two injections into the lower trapezius muscle (preferably one injection into the lower trapezius muscle on a first side of the neck and / or one injection into the lower trapezius muscle on a second side of the neck).

[0333] If the disorder is headache (e.g., migraine pain) or migraine, at least a unit dose of the chimeric Clostridial neurotoxin can be administered to one or more of the frontalis muscle, corrugator muscle, nasal muscle, orbicularis oculi muscle, masseter muscle, temporalis muscle, upper occipital muscle, lower occipital muscle, and upper and lower trapezius muscles. At least a unit dose of the chimeric Clostridial neurotoxin can be administered to the frontalis muscle, corrugator muscle, nasal muscle, orbicularis oculi muscle, masseter muscle, temporalis muscle, upper occipital muscle, lower occipital muscle, and upper and lower trapezius muscles. In one embodiment: (i) a single unit dose is administered to one or more of the frontalis muscle, corrugator muscles (preferably, a single unit dose is administered to the corrugator muscles on a first side of the face (e.g., the left side) and a second unit dose is administered to the corrugator muscles on a second side of the face (e.g., the right side)), nasal muscles, orbicularis oculi muscles, masseter muscles, and lower occipital muscles; and / or (preferably, and) (iii) A plurality of unit doses is administered to one or more of the temporalis, upper occipital, and trapezius muscles. The plurality of unit doses may be 2 to 8 unit doses, for example, 2 to 5 unit doses.

[0334] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 1 unit dose into the frontalis muscle (preferably 1 unit dose per frontalis muscle); (ii) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iii) one unit dose to the bridge of the nose (preferably one unit dose per bridge of the nose); (iv) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (v) 1 unit dose into the masseter muscles (preferably 1 unit dose per masseter muscle); (vi) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (vii) 2 unit doses in the upper occipitalis muscle (preferably 2 unit doses per upper occipitalis muscle); (viii) 1 unit dose into the suboccipitalis muscle (preferably 1 unit dose per suboccipitalis muscle); and / or (viii) 2 unit doses into the trapezius muscle (preferably 2 unit doses per trapezius muscle).

[0335] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) 1 unit dose into the frontalis muscle (preferably 1 unit dose per frontalis muscle); (ii) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (iii) one unit dose to the bridge of the nose (preferably one unit dose per bridge of the nose); (iv) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (v) 1 unit dose into the masseter muscles (preferably 1 unit dose per masseter muscle); (vi) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (vii) 2 unit doses in the upper occipitalis muscle (preferably 2 unit doses per upper occipitalis muscle); (viii) 1 unit dose into the suboccipitalis muscle (preferably 1 unit dose per suboccipitalis muscle); and (viii) 2 unit doses into the trapezius muscle (preferably 2 unit doses per trapezius muscle).

[0336] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) two unit doses to the frontalis muscle (preferably one unit dose to the frontalis muscle on a first side of the face and one unit dose to the frontalis muscle on a second side of the face); (ii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iii) two unit doses to the bridge of the nose (preferably one unit dose to the bridge of the nose on a first side of the face and one unit dose to the bridge of the nose on a second side of the face); (iv) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (v) two unit doses to the masseter muscles (preferably one unit dose to the masseter muscles on a first side of the face and one unit dose to the masseter muscles on a second side of the face); (vi) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (vii) 4 unit doses in the upper occipitalis muscle (preferably 2 unit doses in the upper occipitalis muscle on a first side of the head and 2 unit doses in the upper occipitalis muscle on a second side of the head); (viii) two unit doses in the suboccipitalis muscle (preferably one unit dose in the suboccipitalis muscle on a first side of the head and one unit dose in the suboccipitalis muscle on a second side of the head); and / or (ix) 4 unit doses to the trapezius muscle (preferably 2 unit doses to the trapezius muscle on a first neck side and 2 unit doses to the trapezius muscle on a second neck side).

[0337] Treatment of headache (e.g., migraine pain) or migraine headache may include administration of: (i) two unit doses to the frontalis muscle (preferably one unit dose to the frontalis muscle on a first side of the face and one unit dose to the frontalis muscle on a second side of the face); (ii) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (iii) two unit doses to the bridge of the nose (preferably one unit dose to the bridge of the nose on a first side of the face and one unit dose to the bridge of the nose on a second side of the face); (iv) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (v) two unit doses to the masseter muscles (preferably one unit dose to the masseter muscles on a first side of the face and one unit dose to the masseter muscles on a second side of the face); (vi) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (vii) 4 unit doses in the upper occipitalis muscle (preferably 2 unit doses in the upper occipitalis muscle on a first side of the head and 2 unit doses in the upper occipitalis muscle on a second side of the head); (viii) two unit doses in the suboccipitalis muscle (preferably one unit dose in the suboccipitalis muscle on a first side of the head and one unit dose in the suboccipitalis muscle on a second side of the head); and (ix) 4 unit doses to the trapezius muscle (preferably 2 unit doses to the trapezius muscle on a first neck side and 2 unit doses to the trapezius muscle on a second neck side).

[0338] Administration of the chimeric Clostridial neurotoxin may include: (i) one injection into the frontalis muscle (preferably one injection per frontalis muscle); (ii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iii) a single injection into the bridge of the nose (preferably one injection per bridge of the nose); (iv) one injection into the orbicularis oculi muscle (preferably one injection per orbicularis oculi muscle); (v) one injection into the masseter muscles (preferably one injection per masseter muscle); (vi) four injections into the temporalis muscle (preferably four injections per temporalis muscle); (vii) two injections into the superior occipitalis muscle (preferably two injections into each superior occipitalis muscle); (viii) one injection into the suboccipitalis muscle (preferably one injection per suboccipitalis muscle); and / or (viii) Two injections into the trapezius muscle (preferably two injections per trapezius muscle).

[0339] Administration of the chimeric Clostridial neurotoxin may include: (i) one injection into the frontalis muscle (preferably one injection per frontalis muscle); (ii) a single injection into the corrugator muscles (preferably one injection per corrugator muscle); (iii) a single injection into the bridge of the nose (preferably one injection per bridge of the nose); (iv) one injection into the orbicularis oculi muscle (preferably one injection per orbicularis oculi muscle); (v) one injection into the masseter muscles (preferably one injection per masseter muscle); (vi) four injections into the temporalis muscle (preferably four injections per temporalis muscle); (vii) two injections into the superior occipitalis muscle (preferably two injections into each superior occipitalis muscle); (viii) one injection into the suboccipitalis muscle (preferably one injection per suboccipitalis muscle); and (viii) Two injections into the trapezius muscle (preferably two injections per trapezius muscle).

[0340] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably one injection into the frontalis muscle on a first side of the face and one injection into the frontalis muscle on a second side of the face); (ii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iii) two injections into the nasal bridge (preferably one injection into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face); (iv) two injections into the orbicularis oculi muscle (preferably one injection into the orbicularis oculi muscle on a first side of the face and one injection into the orbicularis oculi muscle on a second side of the face); (v) two injections into the masseter muscles (preferably one injection into the masseter muscles on a first side of the face and one injection into the masseter muscles on a second side of the face); (vi) eight injections into the temporalis muscle (preferably four injections into the temporalis muscle on the first side of the head and four injections into the temporalis muscle on the second side of the head); (vii) four injections into the upper occipitalis muscle (preferably two injections into the upper occipitalis muscle on the first side of the head and two injections into the upper occipitalis muscle on the second side of the head); (viii) two injections into the suboccipitalis muscle (preferably one injection into the suboccipitalis muscle on a first side of the head and one injection into the suboccipitalis muscle on a second side of the head); and / or (ix) Four injections into the trapezius muscle (preferably two injections into the trapezius muscle on the first side of the neck and two injections into the trapezius muscle on the second side of the neck).

[0341] Administration of the chimeric Clostridial neurotoxin may include: (i) two injections into the frontalis muscle (preferably one injection into the frontalis muscle on a first side of the face and one injection into the frontalis muscle on a second side of the face); (ii) two injections into the corrugator muscles (preferably one injection into the corrugator muscles on a first side of the face and one injection into the corrugator muscles on a second side of the face); (iii) two injections into the nasal bridge (preferably one injection into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face); (iv) two injections into the orbicularis oculi muscle (preferably one injection into the orbicularis oculi muscle on a first side of the face and one injection into the orbicularis oculi muscle on a second side of the face); (v) two injections into the masseter muscles (preferably one injection into the masseter muscles on a first side of the face and one injection into the masseter muscles on a second side of the face); (vi) eight injections into the temporalis muscle (preferably four injections into the temporalis muscle on the first side of the head and four injections into the temporalis muscle on the second side of the head); (vii) four injections into the upper occipitalis muscle (preferably two injections into the upper occipitalis muscle on the first side of the head and two injections into the upper occipitalis muscle on the second side of the head); (viii) two injections into the suboccipitalis muscle (preferably one injection into the suboccipitalis muscle on the first side of the head and one injection into the suboccipitalis muscle on the second side of the head); and (ix) Four injections into the trapezius muscle (preferably two injections into the trapezius muscle on the first side of the neck and two injections into the trapezius muscle on the second side of the neck).

[0342] In any aspect or embodiment described herein, administration of the chimeric Clostridial neurotoxin may include direct or indirect injection of the chimeric Clostridial neurotoxin into a muscle. For example, when the chimeric Clostridial neurotoxin is injected indirectly into a muscle, the chimeric Clostridial neurotoxin may be administered to the muscle region. In one embodiment, when the chimeric Clostridial neurotoxin is injected directly into a muscle, the chimeric Clostridial neurotoxin may be administered intramuscularly into the muscle. In one embodiment, when the chimeric Clostridial neurotoxin is injected indirectly into a muscle, the chimeric Clostridial neurotoxin may be administered intradermally.

[0343] If the disorder is headache (e.g., migraine pain) or migraine, at least a unit dose of the chimeric Clostridial neurotoxin can be administered intradermally to one or more of the ophthalmic region of the trigeminal nerve, the maxillary region of the trigeminal nerve, the mandibular region of the trigeminal nerve, and the occipital region. Preferably, at least a unit dose of the chimeric Clostridial neurotoxin can be administered intradermally to the ophthalmic region of the trigeminal nerve, the maxillary region of the trigeminal nerve, the mandibular region of the trigeminal nerve, and the occipital region.

[0344] Intradermal administration in one or more of the above regions can target the chimeric Clostridial neurotoxin to the target trigeminal nerve (e.g., target nerve ending). The target nerve (e.g., target nerve ending) in the ophthalmic region of the trigeminal nerve can be one or more of the supraorbital nerve, supratrochlear nerve, and infratrochlear nerve (e.g., their nerve endings). The target nerve (e.g., target nerve ending) in the maxillary region of the trigeminal nerve can be one or more of the zygomaticotemporal nerve and the zygomaticofacial nerve (e.g., their nerve endings). The target nerve (e.g., target nerve ending) in the mandibular region of the trigeminal nerve can be the auriculotemporal nerve (e.g., its nerve ending). The target nerve (e.g., target nerve ending) in the occipital region can be one or more of the greater occipital nerve and the lesser occipital nerve (e.g., their nerve endings).

[0345] Intradermal administration in one or more of the above regions can target the chimeric Clostridial neurotoxin to the target trigeminal nerve (e.g., target nerve ending). The target nerve (e.g., target nerve ending) in the ophthalmic region of the trigeminal nerve can be one or more of the supraorbital nerve and the supratrochlear nerve (e.g., their nerve endings). The target nerve (e.g., target nerve ending) in the maxillary region of the trigeminal nerve can be one or more of the zygomaticotemporal nerve and the intraorbital nerve (e.g., their nerve endings). The target nerve (e.g., target nerve ending) in the mandibular region of the trigeminal nerve can be one or more of the auriculotemporal nerve and the mandibular nerve (e.g., their nerve endings). The target nerve (e.g., target nerve ending) in the occipital region can be one or more of the greater occipital nerve, the lesser occipital nerve, and the suboccipital nerve (e.g., their nerve endings).

[0346] In one embodiment, (i) A single unit dose is administered intradermally to one or more of the following regions: the supraorbital nerve (preferably, a single unit dose is administered to the supraorbital nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the supraorbital nerve region of a second facial side (e.g., right side)); the supratrochlear nerve (preferably, a single unit dose is administered to the supratrochlear nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the supratrochlear nerve region of a second facial side (e.g., right side)); the infratrochlear nerve (preferably, a single unit dose is administered to the infratrochlear nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the infratrochlear nerve region of a second facial side (e.g., right side)). the zygomaticotemporal nerve (preferably, a single unit dose is administered to the zygomaticotemporal nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the zygomaticotemporal nerve region of a second facial side (e.g., right side)); the zygomaticofacial nerve (preferably, a single unit dose is administered to the zygomaticotemporal nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the zygomaticotemporal nerve region of a second facial side (e.g., right side)); the lesser occipital nerve (preferably, a single unit dose is administered to the lesser occipital nerve region of a first neck side (e.g., left side) and a second unit dose is administered to the lesser occipital nerve region of a second neck side (e.g., right side); and / or (preferably, and) (iii) Multiple unit doses are administered intradermally to one or more of the following regions: the supraorbital nerve (preferably, a single unit dose is administered to the supraorbital nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the supraorbital nerve region of a second facial side (e.g., right side)); the supratrochlear nerve (preferably, a single unit dose is administered to the supratrochlear nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the supratrochlear nerve region of a second facial side (e.g., right side)); the infratrochlear nerve (preferably, a single unit dose is administered to the infratrochlear nerve region of a first facial side (e.g., left side) and a second unit dose is administered to the infratrochlear nerve region of a second facial side (e.g., right side)). the zygomaticotemporal nerve (preferably, a single unit dose is administered to the zygomaticotemporal nerve region of a first facial side (e.g., the left side), and a second unit dose is administered to the zygomaticotemporal nerve region of a second facial side (e.g., the right side)); the zygomaticofacial nerve (preferably, a single unit dose is administered to the zygomaticotemporal nerve region of a first facial side (e.g., the left side), and a second unit dose is administered to the zygomaticotemporal nerve region of a second facial side (e.g., the right side)); the auriculotemporal nerve; the greater occipital nerve; the lesser occipital nerve (preferably, a single unit dose is administered to the lesser occipital nerve region of a first head side (e.g., the left side), and a second unit dose is administered to the lesser occipital nerve region of a second head side (e.g., the right side)). The multiple unit doses may be 2 to 8 unit doses, for example, 2 to 5 unit doses.

[0347] Preferred injection sites and times are shown in Figure 6. In such instances, one unit dose of chimeric Clostridial neurotoxin may be administered per injection site.

[0348] Preferably, the injection site is within the terminal area of ​​the indicated nerve.

[0349] Treatment of headache (e.g., migraine pain) or migraine headache can include bilateral intradermal administration of a unit dose of a chimeric Clostridial neurotoxin. Treatment of headache (e.g., migraine pain) or migraine headache can include administration of: (i) one unit dose in the supraorbital nerve region of a first side of the face and / or one unit dose in the supraorbital nerve region of a second side of the face; (ii) one unit dose in the region of the supratrochlear nerve on a first side of the face and / or one unit dose in the region of the supratrochlear nerve on a second side of the face; (iii) one unit dose in the region of the infratrochlear nerve on a first side of the face and / or one unit dose in the region of the infratrochlear nerve on a second side of the face; (iv) one unit dose in the zygomaticotemporal nerve region of a first side of the face and / or one unit dose in the zygomaticotemporal nerve region of a second side of the face; (v) one unit dose in the zygomaticofacial nerve region of a first side of the face and / or one unit dose in the zygomaticofacial nerve region of a second side of the face; (vi) 2 unit doses to the auriculotemporal nerve region of a first side of the face and / or 2 unit doses to the auriculotemporal nerve region of a second side of the face; (vii) 2 unit doses in the area of ​​the greater occipital nerve on a first side of the neck, and / or 2 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) 1 unit dose in the area of ​​the lesser occipital nerve on a first side of the neck and / or 1 unit dose in the area of ​​the lesser occipital nerve on a second side of the neck.

[0350] Treatment of headache (e.g., migraine pain) or migraine headache can include administering a unit dose of a chimeric Clostridial neurotoxin bilaterally. Treatment of headache (e.g., migraine pain) or migraine headache can include administering: (i) one unit dose in the supraorbital nerve region of a first side of the face and / or one unit dose in the supraorbital nerve region of a second side of the face; (ii) one unit dose in the region of the supratrochlear nerve on a first side of the face and / or one unit dose in the region of the supratrochlear nerve on a second side of the face; (iii) one unit dose in the region of the infratrochlear nerve on a first side of the face and / or one unit dose in the region of the infratrochlear nerve on a second side of the face; (iv) one unit dose in the zygomaticotemporal nerve region of a first side of the face and / or one unit dose in the zygomaticotemporal nerve region of a second side of the face; (v) one unit dose in the zygomaticofacial nerve region of a first side of the face and / or one unit dose in the zygomaticofacial nerve region of a second side of the face; (vi) 2 unit doses to the auriculotemporal nerve region of a first side of the face and / or 2 unit doses to the auriculotemporal nerve region of a second side of the face; (vii) 2 unit doses in the area of ​​the greater occipital nerve on a first side of the neck, and / or 2 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) 1 unit dose in the area of ​​the lesser occipital nerve on a first side of the neck and / or 1 unit dose in the area of ​​the lesser occipital nerve on a second side of the neck.

[0351] Preferably, treatment of headache (e.g., migraine pain) or migraine may include administration of: (i) 1 unit dose in the supraorbital nerve region of a first side of the face and 1 unit dose in the supraorbital nerve region of a second side of the face; (ii) one unit dose in the region of the supratrochlear nerve on a first side of the face and one unit dose in the region of the supratrochlear nerve on a second side of the face; (iii) one unit dose in the region of the infratrochlear nerve on a first side of the face and one unit dose in the region of the infratrochlear nerve on a second side of the face; (iv) 1 unit dose in the zygomaticotemporal nerve region of the first side of the face and 1 unit dose in the zygomaticotemporal nerve region of the second side of the face; (v) one unit dose in the zygomaticofacial nerve region of the first side of the face and one unit dose in the zygomaticofacial nerve region of the second side of the face; (vi) 2 unit doses in the auriculotemporal nerve region of a first side of the face and 2 unit doses in the auriculotemporal nerve region of a second side of the face; (vii) 2 unit doses in the area of ​​the greater occipital nerve on a first side of the neck and 2 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) 1 unit dose in the area of ​​the lesser occipital nerve on a first side of the neck and 1 unit dose in the area of ​​the lesser occipital nerve on a second side of the neck.

[0352] More preferably, treatment of headache (e.g., migraine pain) or migraine may include administration of: (i) 1 unit dose in the supraorbital nerve region of a first side of the face and 1 unit dose in the supraorbital nerve region of a second side of the face; (ii) one unit dose in the region of the supratrochlear nerve on a first side of the face and one unit dose in the region of the supratrochlear nerve on a second side of the face; (iii) one unit dose in the region of the infratrochlear nerve on a first side of the face and one unit dose in the region of the infratrochlear nerve on a second side of the face; (iv) 1 unit dose in the zygomaticotemporal nerve region of the first side of the face and 1 unit dose in the zygomaticotemporal nerve region of the second side of the face; (v) one unit dose in the zygomaticofacial nerve region of the first side of the face and one unit dose in the zygomaticofacial nerve region of the second side of the face; (vi) 2 unit doses in the auriculotemporal nerve region of a first side of the face and 2 unit doses in the auriculotemporal nerve region of a second side of the face; (vii) 2 unit doses in the area of ​​the greater occipital nerve on a first side of the neck and 2 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and (vii) 1 unit dose in the area of ​​the lesser occipital nerve on a first side of the neck and 1 unit dose in the area of ​​the lesser occipital nerve on a second side of the neck.

[0353] Treatment of headache (e.g., migraine pain) or migraine headache can include bilateral intradermal administration of a unit dose of a chimeric Clostridial neurotoxin. Treatment of headache (e.g., migraine pain) or migraine headache can include administration of: (i) 2 unit doses in the supraorbital nerve region of a first side of the face and / or 2 unit doses in the supraorbital nerve region of a second side of the face; (ii) 2 unit doses in the region of the supratrochlear nerve on a first side of the face and / or 2 unit doses in the region of the supratrochlear nerve on a second side of the face; (iii) 2 unit doses in the area of ​​the infratrochlear nerve on a first side of the face and / or 2 unit doses in the area of ​​the infratrochlear nerve on a second side of the face; (iv) 2 unit doses in the zygomaticotemporal nerve region of the first side of the face and / or 2 unit doses in the zygomaticotemporal nerve region of the second side of the face; (v) 2 unit doses in the zygomaticofacial nerve region of the first side of the face and / or 2 unit doses in the zygomaticofacial nerve region of the second side of the face; (vi) 4 unit doses to the auriculotemporal nerve region of a first side of the face and / or 4 unit doses to the auriculotemporal nerve region of a second side of the face; (vii) 4 unit doses in the area of ​​the greater occipital nerve on a first side of the neck, and / or 4 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) 2 unit doses in the area of ​​the lesser occipital nerve on a first side of the neck and / or 2 unit doses in the area of ​​the lesser occipital nerve on a second side of the neck.

[0354] Treatment of headache (e.g., migraine pain) or migraine headache can include administering a unit dose of a chimeric Clostridial neurotoxin bilaterally. Treatment of headache (e.g., migraine pain) or migraine headache can include administering: (i) 2 unit doses in the supraorbital nerve region of a first side of the face and / or 2 unit doses in the supraorbital nerve region of a second side of the face; (ii) 2 unit doses in the region of the supratrochlear nerve on a first side of the face and / or 2 unit doses in the region of the supratrochlear nerve on a second side of the face; (iii) 2 unit doses in the area of ​​the infratrochlear nerve on a first side of the face and / or 2 unit doses in the area of ​​the infratrochlear nerve on a second side of the face; (iv) 2 unit doses in the zygomaticotemporal nerve region of the first side of the face and / or 2 unit doses in the zygomaticotemporal nerve region of the second side of the face; (v) 2 unit doses in the zygomaticofacial nerve region of the first side of the face and / or 2 unit doses in the zygomaticofacial nerve region of the second side of the face; (vi) 4 unit doses to the auriculotemporal nerve region of a first side of the face and / or 4 unit doses to the auriculotemporal nerve region of a second side of the face; (vii) 4 unit doses in the area of ​​the greater occipital nerve on a first side of the neck, and / or 4 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) 2 unit doses in the area of ​​the lesser occipital nerve on a first side of the neck and / or 2 unit doses in the area of ​​the lesser occipital nerve on a second side of the neck.

[0355] Preferably, treatment of headache (e.g., migraine pain) or migraine may include administration of: (i) 2 unit doses in the supraorbital nerve region of the first side of the face and 2 unit doses in the supraorbital nerve region of the second side of the face; (ii) 2 unit doses in the supratrochlear nerve region of the first side of the face and 2 unit doses in the supratrochlear nerve region of the second side of the face; (iii) 2 unit doses in the area of ​​the infratrochlear nerve on the first side of the face and 2 unit doses in the area of ​​the infratrochlear nerve on the second side of the face; (iv) 2 unit doses in the zygomaticotemporal nerve region of the first facial side and 2 unit doses in the zygomaticotemporal nerve region of the second facial side; (v) 2 unit doses in the zygomaticofacial nerve region of the first side of the face and 2 unit doses in the zygomaticofacial nerve region of the second side of the face; (vi) 4 unit doses in the auriculotemporal nerve region of the first side of the face and 4 unit doses in the auriculotemporal nerve region of the second side of the face; (vii) 4 unit doses in the area of ​​the greater occipital nerve on a first side of the neck and 4 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) 2 unit doses in the area of ​​the lesser occipital nerve on a first side of the neck and 2 unit doses in the area of ​​the lesser occipital nerve on a second side of the neck.

[0356] More preferably, treatment of headache (e.g., migraine pain) or migraine may include administration of: (i) 2 unit doses in the supraorbital nerve region of the first side of the face and 2 unit doses in the supraorbital nerve region of the second side of the face; (ii) 2 unit doses in the supratrochlear nerve region of the first side of the face and 2 unit doses in the supratrochlear nerve region of the second side of the face; (iii) 2 unit doses in the area of ​​the infratrochlear nerve on the first side of the face and 2 unit doses in the area of ​​the infratrochlear nerve on the second side of the face; (iv) 2 unit doses in the zygomaticotemporal nerve region of the first facial side and 2 unit doses in the zygomaticotemporal nerve region of the second facial side; (v) 2 unit doses in the zygomaticofacial nerve region of the first side of the face and 2 unit doses in the zygomaticofacial nerve region of the second side of the face; (vi) 4 unit doses in the auriculotemporal nerve region of the first side of the face and 4 unit doses in the auriculotemporal nerve region of the second side of the face; (vii) 4 unit doses in the area of ​​the greater occipital nerve on a first side of the neck and 4 unit doses in the area of ​​the greater occipital nerve on a second side of the neck; and (vii) 2 unit doses in the area of ​​the lesser occipital nerve on a first side of the neck and 2 unit doses in the area of ​​the lesser occipital nerve on a second side of the neck.

[0357] When treating headache (e.g., migraine pain) or migraine headache as described in the above embodiments, it is preferred to administer one unit dose per injection site. Thus, treatment can include administering a chimeric Clostridial neurotoxin to: (i) one injection site in the supraorbital nerve region on a first side of the face and / or one injection site in the supraorbital nerve region on a second side of the face; (ii) one injection site in the supratrochlear nerve region on a first side of the face and / or one injection site in the supratrochlear nerve region on a second side of the face; (iii) one injection site in the region of the infratrochlear nerve on a first side of the face and / or one injection site in the region of the infratrochlear nerve on a second side of the face; (iv) one injection site in the zygomaticotemporal nerve region on a first side of the face and / or one injection site in the zygomaticotemporal nerve region on a second side of the face; (v) one injection site in the zygomaticofacial nerve region on a first side of the face and / or one injection site in the zygomaticofacial nerve region on a second side of the face; (vi) two injection sites in the auriculotemporal nerve region on a first side of the face and / or two injection sites in the auriculotemporal nerve region on a second side of the face; (vii) two injection sites in the area of ​​the greater occipital nerve on a first side of the neck and / or two injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) One injection site in the area of ​​the lesser occipital nerve on a first side of the neck and / or one injection site in the area of ​​the lesser occipital nerve on a second side of the neck.

[0358] Preferably, the treatment may involve administering a chimeric Clostridial neurotoxin to: (i) one injection site in the supraorbital nerve region on a first side of the face and one injection site in the supraorbital nerve region on a second side of the face; (ii) one injection site in the supratrochlear nerve region on a first side of the face and one injection site in the supratrochlear nerve region on a second side of the face; (iii) one injection site in the infratrochlear nerve region on a first side of the face and one injection site in the infratrochlear nerve region on a second side of the face; (iv) one injection site in the zygomaticotemporal nerve region on the first side of the face and one injection site in the zygomaticotemporal nerve region on the second side of the face; (v) one injection site in the zygomaticofacial nerve region on the first side of the face and one injection site in the zygomaticofacial nerve region on the second side of the face; (vi) two injection sites in the auriculotemporal nerve region on the first side of the face and two injection sites in the auriculotemporal nerve region on the second side of the face; (vii) two injection sites in the area of ​​the greater occipital nerve on a first side of the neck and two injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) One injection site in the area of ​​the lesser occipital nerve on a first side of the neck and one injection site in the area of ​​the lesser occipital nerve on a second side of the neck.

[0359] More preferably, the treatment may involve administering a chimeric Clostridial neurotoxin to: (i) one injection site in the supraorbital nerve region on a first side of the face and one injection site in the supraorbital nerve region on a second side of the face; (ii) one injection site in the supratrochlear nerve region on a first side of the face and one injection site in the supratrochlear nerve region on a second side of the face; (iii) one injection site in the infratrochlear nerve region on a first side of the face and one injection site in the infratrochlear nerve region on a second side of the face; (iv) one injection site in the zygomaticotemporal nerve region on the first side of the face and one injection site in the zygomaticotemporal nerve region on the second side of the face; (v) one injection site in the zygomaticofacial nerve region on the first side of the face and one injection site in the zygomaticofacial nerve region on the second side of the face; (vi) two injection sites in the auriculotemporal nerve region on the first side of the face and two injection sites in the auriculotemporal nerve region on the second side of the face; (vii) two injection sites in the area of ​​the greater occipital nerve on the first side of the neck and two injection sites in the area of ​​the greater occipital nerve on the second side of the neck; and (vii) One injection site in the area of ​​the lesser occipital nerve on a first side of the neck and one injection site in the area of ​​the lesser occipital nerve on a second side of the neck.

[0360] When treating headache (e.g., migraine pain) or migraine headache as described in the above embodiments, it is preferred to administer one unit dose per injection site. Thus, treatment can include administering a chimeric Clostridial neurotoxin to: (i) two injection sites in the supraorbital nerve region on a first side of the face and / or two injection sites in the supraorbital nerve region on a second side of the face; (ii) two injection sites in the supratrochlear nerve region on a first side of the face and / or two injection sites in the supratrochlear nerve region on a second side of the face; (iii) two injection sites in the infratrochlear nerve region on a first side of the face and / or two injection sites in the infratrochlear nerve region on a second side of the face; (iv) two injection sites in the zygomaticotemporal nerve region on a first side of the face and / or two injection sites in the zygomaticotemporal nerve region on a second side of the face; (v) two injection sites in the zygomaticofacial nerve region on a first side of the face and / or two injection sites in the zygomaticofacial nerve region on a second side of the face; (vi) four injection sites in the auriculotemporal nerve region on a first side of the face and / or four injection sites in the auriculotemporal nerve region on a second side of the face; (vii) four injection sites in the area of ​​the greater occipital nerve on a first side of the neck and / or four injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) Two injection sites in the area of ​​the lesser occipital nerve on a first side of the neck and / or two injection sites in the area of ​​the lesser occipital nerve on a second side of the neck.

[0361] Preferably, the treatment may involve administering a chimeric Clostridial neurotoxin to: (i) two injection sites in the supraorbital nerve region on the first side of the face and two injection sites in the supraorbital nerve region on the second side of the face; (ii) two injection sites in the supratrochlear nerve region on the first side of the face and two injection sites in the supratrochlear nerve region on the second side of the face; (iii) two injection sites in the infratrochlear nerve territory of the first facial side and two injection sites in the infratrochlear nerve territory of the second facial side; (iv) two injection sites in the zygomaticotemporal nerve region on the first facial side and two injection sites in the zygomaticotemporal nerve region on the second facial side; (v) two injection sites in the zygomaticofacial nerve region on the first side of the face and two injection sites in the zygomaticofacial nerve region on the second side of the face; (vi) four injection sites in the auriculotemporal nerve region on the first side of the face and four injection sites in the auriculotemporal nerve region on the second side of the face; (vii) four injection sites in the area of ​​the greater occipital nerve on a first side of the neck and four injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) Two injection sites in the area of ​​the lesser occipital nerve on the first side of the neck and two injection sites in the area of ​​the lesser occipital nerve on the second side of the neck.

[0362] More preferably, the treatment may involve administering a chimeric Clostridial neurotoxin to: (i) two injection sites in the supraorbital nerve region on the first side of the face and two injection sites in the supraorbital nerve region on the second side of the face; (ii) two injection sites in the supratrochlear nerve region on the first side of the face and two injection sites in the supratrochlear nerve region on the second side of the face; (iii) two injection sites in the infratrochlear nerve territory of the first facial side and two injection sites in the infratrochlear nerve territory of the second facial side; (iv) two injection sites in the zygomaticotemporal nerve region on the first facial side and two injection sites in the zygomaticotemporal nerve region on the second facial side; (v) two injection sites in the zygomaticofacial nerve region on the first side of the face and two injection sites in the zygomaticofacial nerve region on the second side of the face; (vi) four injection sites in the auriculotemporal nerve region on the first side of the face and four injection sites in the auriculotemporal nerve region on the second side of the face; (vii) four injection sites in the area of ​​the greater occipital nerve on a first side of the neck and four injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and (vii) Two injection sites in the area of ​​the lesser occipital nerve on the first side of the neck and two injection sites in the area of ​​the lesser occipital nerve on the second side of the neck.

[0363] When treating headache (e.g., migraine pain) or migraine headache as described in the above embodiments, it is preferred to administer two or more unit doses (preferably two unit doses) per injection site. Thus, treatment can include administering a chimeric Clostridial neurotoxin to: (i) one injection site in the supraorbital nerve region on a first side of the face and / or one injection site in the supraorbital nerve region on a second side of the face; (ii) one injection site in the supratrochlear nerve region on a first side of the face and / or one injection site in the supratrochlear nerve region on a second side of the face; (iii) one injection site in the region of the infratrochlear nerve on a first side of the face and / or one injection site in the region of the infratrochlear nerve on a second side of the face; (iv) one injection site in the zygomaticotemporal nerve region on a first side of the face and / or one injection site in the zygomaticotemporal nerve region on a second side of the face; (v) one injection site in the zygomaticofacial nerve region on a first side of the face and / or one injection site in the zygomaticofacial nerve region on a second side of the face; (vi) two injection sites in the auriculotemporal nerve region on a first side of the face and / or two injection sites in the auriculotemporal nerve region on a second side of the face; (vii) two injection sites in the area of ​​the greater occipital nerve on a first side of the neck and / or two injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) One injection site in the area of ​​the lesser occipital nerve on a first side of the neck and / or one injection site in the area of ​​the lesser occipital nerve on a second side of the neck.

[0364] Preferably, the treatment may involve administering a chimeric Clostridial neurotoxin to: (i) one injection site in the supraorbital nerve region on a first side of the face and one injection site in the supraorbital nerve region on a second side of the face; (ii) one injection site in the supratrochlear nerve region on a first side of the face and one injection site in the supratrochlear nerve region on a second side of the face; (iii) one injection site in the infratrochlear nerve region on a first side of the face and one injection site in the infratrochlear nerve region on a second side of the face; (iv) one injection site in the zygomaticotemporal nerve region on the first side of the face and one injection site in the zygomaticotemporal nerve region on the second side of the face; (v) one injection site in the zygomaticofacial nerve region on the first side of the face and one injection site in the zygomaticofacial nerve region on the second side of the face; (vi) two injection sites in the auriculotemporal nerve region on the first side of the face and two injection sites in the auriculotemporal nerve region on the second side of the face; (vii) two injection sites in the area of ​​the greater occipital nerve on a first side of the neck and two injection sites in the area of ​​the greater occipital nerve on a second side of the neck; and / or (vii) One injection site in the area of ​​the lesser occipital nerve on a first side of the neck and one injection site in the area of ​​the lesser occipital nerve on a second side of the neck.

[0365] More preferably, the treatment may involve administering a chimeric Clostridial neurotoxin to: (i) one injection site in the supraorbital nerve region on a first side of the face and one injection site in the supraorbital nerve region on a second side of the face; (ii) one injection site in the supratrochlear nerve region on a first side of the face and one injection site in the supratrochlear nerve region on a second side of the face; (iii) one injection site in the infratrochlear nerve region on a first side of the face and one injection site in the infratrochlear nerve region on a second side of the face; (iv) one injection site in the zygomaticotemporal nerve region on the first side of the face and one injection site in the zygomaticotemporal nerve region on the second side of the face; (v) one injection site in the zygomaticofacial nerve region on the first side of the face and one injection site in the zygomaticofacial nerve region on the second side of the face; (vi) two injection sites in the auriculotemporal nerve region on the first side of the face and two injection sites in the auriculotemporal nerve region on the second side of the face; (vii) two injection sites in the area of ​​the greater occipital nerve on the first side of the neck and two injection sites in the area of ​​the greater occipital nerve on the second side of the neck; and (vii) One injection site in the area of ​​the lesser occipital nerve on a first side of the neck and one injection site in the area of ​​the lesser occipital nerve on a second side of the neck.

[0366] Thus, when treating headache (e.g., migraine pain) or migraine, 1 to 50, 5 to 45, or 10 to 38 unit doses may be administered. Preferably, up to 35 unit doses are administered. Preferably, the total dose administered per treatment session may be up to 192,500 pg of chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 180,000 pg, preferably up to 177,000 pg (more preferably up to 175,000 pg). Most preferably, the total dose administered may be up to 115,000 pg or 75,000 pg, e.g., up to 112,000 pg or 70,000 pg.

[0367] Thus, when treating headache (e.g., migraine pain) or migraine headache via intramuscular injection, 1 to 50, 5 to 45, or 10 to 38 unit doses may be administered. Preferably, up to 35 unit doses are administered. Preferably, the total dose administered per treatment session may be up to 192,500 pg of chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 180,000 pg, preferably up to 177,000 pg (more preferably up to 175,000 pg). Most preferably, the total dose administered may be up to 115,000 pg or 75,000 pg, e.g., up to 112,000 pg or 70,000 pg.

[0368] Thus, when treating headache (e.g., migraine pain) or migraine headache via intradermal injection, 1 to 35, 5 to 25, or 10 to 20 unit doses may be administered. Preferably, up to 20 unit doses are administered. Preferably, the total dose administered per treatment session may be up to 110,000 pg of chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 105,000 pg, preferably up to 102,000 pg (more preferably up to 100,000 pg). When treating headache (e.g., migraine pain) or migraine headache via intradermal injection, 2 to 70, 10 to 50, or 20 to 40 unit doses may be administered. Preferably, up to 40 unit doses are administered. Preferably, the total dose administered per treatment session may be up to 220,000 pg of chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 210,000 pg, preferably up to 204,000 pg (more preferably up to 200,000 pg).

[0369] In some embodiments, the treatment of headache (e.g., migraine pain) or migraine may be performed by a combination of intramuscular and intradermal injections. For example, the chimeric Clostridial neurotoxin of the present invention may be administered intradermally to the neck of a subject, and intramuscularly to the face of the subject. Preferably, the chimeric Clostridial neurotoxin of the present invention may be administered intradermally to the face of a subject, and intramuscularly to the neck of the subject. The chimeric Clostridial neurotoxin may be administered to the head, for example, in addition to administration to the neck and / or face of the subject.

[0370] A preferred unit dose for treating headache (e.g., migraine pain) or migraine headache via intradermal or intramuscular injection may be 1,000 pg to 5,500 pg, with the upper end of the unit dose range being 5,250, 5,200, 5,100, 5,000, 4,500, 4,000, 3,500, 3,000, 2,500, or 2,000 pg of chimeric Clostridial neurotoxin, with the upper end being preferably 5,100 pg and more preferably 5,000 pg. The lower limit of the unit dose range may be 1,100, 1,200, 1,250, 1,300, 1,350, 1,400, 1,450, 1,500, 2,000, 2,500, 3,000, 3,500, or 4,000 pg of chimeric Clostridial neurotoxin, preferably 1,400 pg, more preferably 1,500 pg. The lower limit of the range may be greater than 3,000 pg. The unit dose may be 1,400 pg to 5,100 pg (e.g., 1,500 pg to 5,000 pg), 2,000 pg to 5,100 pg, 3,000 pg to 5,100 pg, or 3,000 pg to 4,000 pg of chimeric Clostridial neurotoxin. The unit dose can contain more than 3,000 pg, up to 5,500 pg, of the chimeric Clostridial neurotoxin. The unit dose of the chimeric Clostridial neurotoxin can be 2,000 pg to 4,500 pg, 2,000 pg to 3,000 pg (e.g., 2,500 pg), or 3,500 to 4,500 pg of the chimeric Clostridial neurotoxin. Preferably, the unit dose contains 4,000 pg of the chimeric Clostridial neurotoxin.

[0371] A preferred unit dose for treating headache (e.g., migraine pain) or migraine headache via intradermal or intramuscular injection may be 42 to 229 units. The upper end of the unit dose range may be 225, 220, 215, 210, 205, 200, 190, 180, 170, 160, 150, 125, 100, or 83 units of chimeric clostridial neurotoxin, preferably with an upper limit of 212 units and more preferably 208 units. The lower end of the unit dose range may be 46, 50, 55, 60, 65, 70, 75, 80, 90, 100, 110, 120, 130, 140, 150, 160, or 166 units of chimeric clostridial neurotoxin, preferably with a lower limit of 58 units and more preferably 62 units. The lower end of the range may be greater than 125 units. A unit dose may be 58 to 212 units (e.g., 62 to 208 units), 83 to 212 units, 125 to 212 units, or 125 to 166 units of a chimeric Clostridial neurotoxin. A unit dose may contain more than 125 units, up to 229 units of a chimeric Clostridial neurotoxin. A unit dose of a chimeric Clostridial neurotoxin may be 83 to 188 units, 83 to 125 units (e.g., 104 units), or 146 to 188 units of a chimeric Clostridial neurotoxin. Preferably, a unit dose contains 166 units of a chimeric Clostridial neurotoxin. A unit dose may contain 47 to 258 units of a chimeric Clostridial neurotoxin, for example, 94 to 211 units, 94 to 141 units (e.g., 117 units), or 164 to 211 units of a chimeric Clostridial neurotoxin. A unit dosage form can contain 188 units of a chimeric clostridial neurotoxin.

[0372] When treating headache (e.g., migraine pain) or migraine headache via intramuscular or intradermal injection (preferably intramuscular injection), a preferred unit dose may be 2,500 pg, with a total dose of up to 70,000 pg. For example, a preferred unit dose may be 2,500 pg, with a total dose of 70,000 pg. A preferred unit dose may be 4,000 pg, with a total dose of up to 112,000 pg. For example, a preferred unit dose may be 4,000 pg, with a total dose of 112,000 pg. A suitable unit dose may be 5,000 pg, with a total dose of up to 155,000 pg. For example, a suitable unit dose may be 5,000 pg, with a total dose of 155,000 pg.

[0373] When treating headache (e.g., migraine pain) or migraine headache via intramuscular or intradermal injection (preferably intramuscular injection), a preferred unit dose may be 104 units, with a total dose of up to 2,912 units. For example, a preferred unit dose may be 104 units, with a total dose of 2,912 units. A preferred unit dose may be 166 units, with a total dose of up to 4,659 units. For example, a preferred unit dose may be 166 units, with a total dose of 4,659 units. A suitable unit dose may be 208 units, with a total dose of up to 6,448 units. For example, a suitable unit dose may be 208 units, with a total dose of 6,448 units.

[0374] When treating headache (e.g., migraine pain) or migraine headache via intramuscular or intradermal injection (preferably intramuscular injection), a preferred unit dose may be 117 units, with a total dose of up to 3,286 units. For example, a preferred unit dose may be 117 units, with a total dose of 3,286 units. A preferred unit dose may be 188 units, with a total dose of up to 5,258 units. For example, a preferred unit dose may be 188 units, with a total dose of 5,258 units. A suitable unit dose may be 235 units, with a total dose of up to 7,277 units. For example, a suitable unit dose may be 235 units, with a total dose of 7,277 units.

[0375] When treating headache (e.g., migraine pain) or migraine headache via intramuscular injection, the total dose administered per treatment session may be up to 8,007 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 7,488 units, or up to 7,363 units (e.g., up to 7,280 units). Most preferably, the total dose administered may be up to 4,784 units or 3,120 units, e.g., up to 4,659 units or 2,912 units. The total dose administered per treatment session may be up to 9,037 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 8,451 units, or up to 8,310 units (e.g., up to 8,216 units). Most preferably, the total dose administered may be up to 5,399 units or 3,521 units, e.g., up to 5,258 units or 3,286 units.

[0376] When treating headache (e.g., migraine pain) or migraine headache via intradermal injection, the total dose administered per treatment session may be up to 4,576 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 4,368 units, or up to 4,243 units (e.g., up to 4,160 units). The total dose administered per treatment session may be up to 5,165 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 4,929 units, or up to 4,789 units (e.g., up to 4,695 units). The total dose administered per treatment session may be up to 5,165 units of the chimeric Clostridial neurotoxin. For example, the total dose administered may be up to 4,929 units, or up to 4,789 units (e.g., up to 4,695 units).

[0377] In preferred embodiments, when treating pain (e.g., headache or migraine pain) or migraine with a chimeric Clostridial neurotoxin, the treatment does not induce muscle paralysis. For example, in some embodiments, the unit dose of the chimeric Clostridial neurotoxin may be lower than the unit dose of the chimeric Clostridial neurotoxin required to induce muscle paralysis. In particular, in some embodiments, the unit dose of the chimeric Clostridial neurotoxin administered at a particular site (e.g., injection site) may be lower than the unit dose of the chimeric Clostridial neurotoxin required to induce muscle paralysis (e.g., at that site and / or in a muscle).

[0378] The headache is preferably migraine pain, which may be episodic migraine pain or chronic migraine pain, e.g., pain caused by or associated with episodic migraine headaches or pain caused by or associated with chronic migraine headaches.

[0379] The chimeric Clostridial neurotoxins of the present invention are preferably administered repeatedly (e.g., up to 5, 10, 15, or 20 times) as part of a treatment regimen (preferably on different days, e.g., at least every other day during continuous treatment). Repeated administration means administration at least twice, e.g., at least 5, 10, 15, or 20 times. Thus, in one embodiment, the chimeric Clostridial neurotoxins of the present invention can be administered more than once to treat a subject's disorder (preferably, pain). This is particularly appropriate for treating chronic conditions, such as chronic pain, which generally require continuous treatment. In one embodiment, the chimeric Clostridial neurotoxins of the present invention can be administered once a week, twice a month, once a month, every two months, every six months, or once a year, preferably at least twice a year or once a year. In one embodiment, the chimeric Clostridial neurotoxins of the present invention are administered more than once within a period of 10 years, 5 years, 2 years, or 1 year. Preferably, the chimeric Clostridial neurotoxins of the present invention are administered more than once within a period of 1 year. Treatment may continue for at least 6 months, 1 year, 2 years, 3 years, 5 years, 10 years, 15 years, 20 years, 25 years, or 30 years.

[0380] In some embodiments, after an initial administration (e.g., an initial treatment session) of a chimeric Clostridial neurotoxin according to the present invention, the subject may receive a second administration (e.g., a second treatment session) of the chimeric Clostridial neurotoxin. The interval between the first and second administrations may be at least 5, 6, 7, 8, 9, or 10 months. For example, the interval between the first and second administrations may be 5 to 10 months, 5 to 9 months, 5 to 8 months, 6 to 10 months, 6 to 9 months, or 6 to 8 months.

[0381] Preferably, the chimeric Clostridial neurotoxin is not administered with an additional therapeutic or diagnostic agent (e.g., a nucleic acid, protein, peptide, or small molecule therapeutic agent, or a diagnostic agent) in addition to the light and heavy chains. For example, in one embodiment, the chimeric Clostridial neurotoxin is not administered with an additional analgesic agent. In one embodiment, the chimeric Clostridial neurotoxin of the present invention is not administered with a covalently bound therapeutic agent. In one embodiment, the chimeric Clostridial neurotoxin of the present invention is not administered with a non-covalently bound therapeutic agent.

[0382] The chimeric Clostridial neurotoxins are preferably used to treat pain, and can be used to treat subjects suffering from one or more types of pain.

[0383] As used herein, the term "pain" refers to an unpleasant sensory and emotional experience associated with or resembling actual or potential tissue damage, along with any associated physical disturbances that may or may not be apparent to the clinician.

[0384] Pain can be associated with the release of mediators (e.g., neurotransmitters) from neurons. The neurons are preferably neurons to which a chimeric Clostridial neurotoxin of the present invention binds. For example, the mediator can be any mediator involved in pain transmission. The mediator can be a neuropeptide such as substance P, CGRP, or vasoactive intestinal peptide (VIP).

[0385] The mediator may be an inflammatory mediator or a non-inflammatory mediator. The mediator may be one or more of the following: CGRP, neurokinins (e.g., tachykinins, substance P, neurokinin A, neurokinin B, hemokinins, and / or endokinins), adrenocorticotropic hormone (ACTH), glucocorticoids, vasopressin, oxytocin, catecholamines, opioids (e.g., opioid peptides and / or brain opioids), angiotensin II, endorphins, encephalins, vasoactive intestinal peptide (VIP), eicosanoids (e.g., prostaglandins such as prostaglandin E2 (PGE2) and / or leukotrienes), tissue kininogens (e.g., bradykinin), histamines, and / or steroid hormones. ATP, serotonin, potassium, prostacyclin (PGI2), leukotriene B4 (LTB4), nerve growth factor (NGF), protons, ATP, adenosine, 5-hydroxytryptamine (5-HT), histamine, glutamate, norepinephrine (NE), nitric oxide (NO), gamma-aminobutyric acid (GABA), glycine, acetylcholine, cannabinoids, tissue necrosis factor alpha (TNF-α), cytokines (e.g., interleukin (IL)-6, IL-1, and / or IL-8), platelet-activating factor (PAF), neurotrophic growth factor (NGF), glutamate, aspartate, pituitary adenylate cyclase-activating peptide (PACAP), and proteolytic enzymes.

[0386] The mediator may be calcitonin gene-related peptide (CGRP), amylin, pituitary adenylate cyclase-activating peptide (PACAP), oxytocin, neuropeptide Y (NPY), substance P, angiotensin, corticotropin-releasing hormone (CRH), leptin, adiponectin, orexin, and / or melanin-concentrating hormone (MCH).

[0387] The mediator may be one or more selected from CGRP, substance P, and glutamate.

[0388] The mediator may be one or more of a neuropeptide (eg, substance P, CGRP, or VIP), nitric oxide, glutamate, aspartate.

[0389] When the pain is headache (preferably migraine pain), the mediator may be one or more of CGRP, VIP, PACAP, and inflammatory cytokines (e.g., IL-6, IL-8, and / or TNF-α).

[0390] Preferably, the mediator may be CGRP, substance P, and / or an alternative neurokinin.

[0391] Most preferably, the mediator is CGRP. The CGRP may be α-CGRP or β-CGRP, preferably α-CGRP.

[0392] Pain may be associated with the release of pain mediators (e.g., pain neurotransmitters) from neurons containing A-delta or C nerve fibers. The pain mediator may be any pain mediator released / secreted from neurons containing A-delta or C nerve fibers. The pain mediator may be a neuropeptide such as substance P, CGRP, or vasoactive intestinal peptide (VIP).

[0393] The pain mediator may be an inflammatory mediator or a non-inflammatory mediator. The pain mediator may be one or more of the following: CGRP, neurokinins (e.g., tachykinins, substance P, neurokinin A, neurokinin B, hemokinins, and / or endokinins), adrenocorticotropic hormone (ACTH), glucocorticoids, vasopressin, oxytocin, catecholamines, opioids (e.g., opioid peptides and / or brain opioids), angiotensin II, endorphins, encephalins, vasoactive intestinal peptide (VIP), eicosanoids (e.g., prostaglandins such as prostaglandin E2 (PGE2) and / or leukotrienes), tissue kininogens (e.g....

Claims

1. A medicament for use in treating pain, migraine, or sensory disorders in a subject, wherein: The medicament comprises a chimeric Clostridial neurotoxin, and The chimeric clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and botulinum neurotoxin B (BoNT / B) receptor binding domain (H C domain)

2. The pharmaceutical composition of claim 1, wherein the pain is a migraine.

3. The pharmaceutical of claim 1, wherein the subject's pain or sensory disorder is treated for a longer period of time and / or with greater efficacy compared to subjects treated with BoNT / A.

4. The pharmaceutical of claim 1, wherein the subject's pain is more alleviated compared to a subject treated with BoNT / A.

5. The pharmaceutical described in claim 1, wherein the amount of pain mediator in the biological fluids and / or brain of the subject is reduced compared to the amount of the same pain mediator in the same biological fluids and / or brain of a subject treated with BoNT / A.

6. The pharmaceutical composition of claim 1, wherein the pain or sensory disorder is treated by inhibiting the release of mediators from neurons comprising Aδ nerve fibers or C nerve fibers, and wherein the chimeric Clostridial neurotoxin binds to neurons comprising Aδ nerve fibers or C nerve fibers, respectively.

7. The pharmaceutical composition of claim 1, wherein the pain is headache, migraine pain, cluster headache, or bladder pain.

8. The pharmaceutical composition of claim 1 or 7, wherein the pain is migraine pain.

9. The method of claim 1 or 7, wherein the chimeric Clostridial neurotoxin is administered intramuscularly.

10. The method of claim 1 or 7, wherein the chimeric Clostridial neurotoxin is administered in a unit dose of 5 pg to 17,000 pg.

11. 8. The pharmaceutical composition of claim 1 or 7, wherein the total dose administered per treatment session is up to 255,000 pg of the chimeric Clostridial neurotoxin.

12. The method of claim 1 or 7, wherein the chimeric Clostridial neurotoxin is administered to at least one of the following: frontalis muscle, corrugator (e.g., corrugator supercilii), procerus muscle (e.g., procerus nasus), occipitalis muscle (e.g., upper occipitalis or lower occipitalis), temporalis muscle, trapezius muscle (e.g., upper trapezius, middle trapezius, or lower trapezius), masseter muscle, nas muscle, orbicularis oculi muscle, cervical paraspinal muscles, temporalis fascia, superior auricular muscle, anterior auricular muscle, posterior auricular muscle, sternocleidomastoid muscle, platysma muscle, dilator naris anterior muscle, dilator naris posterior muscle, depressor septum nasalis muscle, mentalis muscle, orbicularis oris muscle, zygomaticus muscle, lolis muscle, buccinator muscle, occipitofrontalis muscle, levator labii superior muscle, depressor labii inferior muscle, depressor anguli oris muscle, thyrohyoid muscle, omohyoid muscle, sternohyoid muscle, splenius cervix muscle, splenius capitis muscle, semispinalis cervix muscle, semispinalis capitis muscle, levator scapulae muscle, digastric muscle, or scalene muscle

13. 10. The medicament of claim 1 or 7, wherein the chimeric Clostridial neurotoxin is administered according to: (a) 2 unit doses into the frontalis muscle (preferably 2 unit doses per frontalis muscle); (b) 1 unit dose into the corrugator muscles (preferably 1 unit dose per corrugator muscle); (c) one unit dose into the bridge of the nose (preferably one unit dose per bridge of the nose); (d) 1 unit dose into the orbicularis oculi muscle (preferably 1 unit dose per orbicularis oculi muscle); (e) 4 unit doses into the temporalis muscle (preferably 4 unit doses per temporalis muscle); (f) 3 unit doses into the occipitalis muscle (preferably 3 unit doses per occipitalis muscle); and (g) 2 unit doses into the trapezius muscle (preferably 2 unit doses per trapezius muscle)

14. 10. The medicament of claim 1 or 7, wherein the chimeric Clostridial neurotoxin is administered according to: (a) 4 unit doses to the frontalis muscle (preferably 2 unit doses to the frontalis muscle on a first side of the face and 2 unit doses to the frontalis muscle on a second side of the face); (b) two unit doses to the corrugator muscles (preferably one unit dose to the corrugator muscles on a first side of the face and one unit dose to the corrugator muscles on a second side of the face); (c) two unit doses into the nasal bridge (preferably one unit dose into the nasal bridge on a first side of the face and one injection into the nasal bridge on a second side of the face); (d) two unit doses to the orbicularis oculi muscle (preferably one unit dose to the orbicularis oculi muscle on a first side of the face and one unit dose to the orbicularis oculi muscle on a second side of the face); (e) 8 unit doses to the temporalis muscle (preferably 4 unit doses to the temporalis muscle on the first side of the head and 4 unit doses to the temporalis muscle on the second side of the head); (f) 6 unit doses to the occipitalis muscle (preferably 3 unit doses to the occipitalis muscle on a first side of the head and 3 unit doses to the occipitalis muscle on a second side of the head); and (g) 4 unit doses to the trapezius muscle (preferably 2 unit doses to the trapezius muscle on a first side of the neck and 2 unit doses to the trapezius muscle on a second side of the neck)

15. the pain is a chronic headache, and the chimeric Clostridial neurotoxin treats the chronic headache by inhibiting the release of pain mediators from neurons containing Aδ nerve fibers or C nerve fibers; the chimeric Clostridial neurotoxin binds to neurons containing Aδ nerve fibers or C nerve fibers, respectively; The chimeric clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and botulinum neurotoxin B (BoNT / B) receptor binding domain (H C domain), the light chain exhibits non-cytotoxic protease activity and is capable of cleaving SNARE proteins in the cytosol of target neurons; and The BoNT / BH C The domain contains substitution mutations at E1191M and S1199Y as compared to the unmodified BoNT / B shown as SEQ ID NO: 7, with the numbering of amino acid residues determined by alignment with SEQ ID NO:

7. The pharmaceutical composition according to claim 1 or 7.

16. the pain is an acute headache, and the chimeric Clostridial neurotoxin treats the acute headache by inhibiting the release of pain mediators from neurons containing Aδ nerve fibers or C nerve fibers; the chimeric Clostridial neurotoxin binds to neurons containing Aδ nerve fibers or C nerve fibers, respectively; The chimeric clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and botulinum neurotoxin B (BoNT / B) receptor binding domain (H C domain), the light chain exhibits non-cytotoxic protease activity and is capable of cleaving SNARE proteins in the cytosol of target neurons; and The BoNT / BH C The domain contains substitution mutations at E1191M and S1199Y as compared to the unmodified BoNT / B shown as SEQ ID NO: 7, with the numbering of amino acid residues determined by alignment with SEQ ID NO:

7. The pharmaceutical composition according to claim 1 or 7.

17. A two-chain chimeric Clostridial neurotoxin comprising a light chain comprising SEQ ID NO:17 and a heavy chain comprising SEQ ID NO:

19.

18. 18. The two-chain chimeric Clostridial neurotoxin of claim 17, wherein the light chain and the heavy chain are formed by cysteine ​​residue 429 of SEQ ID NO: 17 or 18 and cysteine ​​residue 6 of SEQ ID NO: 19 and / or are linked to each other by a disulfide bond therebetween.

19. 19. The two-chain chimeric Clostridial neurotoxin of claim 17 or 18, wherein the light chain has SEQ ID NO: 17 and the heavy chain has SEQ ID NO:

19.

20. 19. The two-chain chimeric Clostridial neurotoxin of claim 17 or 18, wherein: The two-chain chimeric Clostridial neurotoxin consists of a light chain of SEQ ID NO: 17 and a heavy chain of SEQ ID NO: 19, and The light chain and the heavy chain are linked to each other by a disulfide bond formed by and / or between cysteine ​​residue 429 of SEQ ID NO: 17 and cysteine ​​residue 6 of SEQ ID NO:

19.

21. 18. A pharmaceutical composition comprising the two-chain chimeric Clostridial neurotoxin of claim 17.

22. A unit dosage form (e.g., for treating pain) comprising: a. 5 pg to 17,000 pg of a chimeric Clostridial neurotoxin; or b. 0.2 units up to 707 units of a chimeric Clostridial neurotoxin, where 1 unit is the calculated median lethal dose (LD) in mice. 50 ) the amount of said chimeric Clostridial neurotoxin equivalent to c. optionally, pharmaceutically acceptable carriers, excipients, adjuvants, and / or salts; wherein the chimeric Clostridial neurotoxin comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N domain), and botulinum neurotoxin B (BoNT / B) receptor binding domain (H C domain)

23. Kit including: (a) the two-chain chimeric Clostridial neurotoxin of claim 17 or 18, the pharmaceutical composition of claim 21, or the unit dosage form of claim 22; and (b) instructions for using the kit, e.g., in treating pain; and (c) optionally, a diluent

24. 23. A medicament for use in treating pain, migraine, or a sensory disorder in a subject, comprising the two-chain chimeric Clostridial neurotoxin of claim 17 or 18, the pharmaceutical composition of claim 21, or the unit dosage form of claim 22.

25. A method for determining whether a Clostridial neurotoxin is suitable for treating pain, said method comprising: (a) comparing the level of calcitonin gene-related peptide (CGRP) in a first sample with the level of CGRP in a second sample, wherein the first sample is obtained from the subject prior to administration of the clostridial neurotoxin, and wherein the second sample is obtained from the same subject after administration of the clostridial neurotoxin; and (b) determining that the clostridial neurotoxin is suitable for treating pain if the level of CGRP in the second sample is lower than the level of CGRP in the first sample; or (c) determining that the clostridial neurotoxin is unsuitable for treating pain if the level of CGRP in the second sample is not lower than (e.g., higher than or the same as) the level of CGRP in the first sample.

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