BoNT / A for treating facial dystonia

Abstract

The present invention is directed to a modified botulinum neurotoxin A (BoNT / A) for use in treating facial dystonia, including blepharospasm and hemifacial spasm, wherein the unit dose of the modified BoNT / A is at least 240 pg of the modified BoNT / A, the modified BoNT / A is administered by intramuscular injection into an affected muscle of a patient, and the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of the modified BoNT / A, the modified BoNT / A comprising a BoNT / A light chain and translocation domain (HN domain) and a BoNT / B receptor binding domain (HC domain). Related methods, uses, unit dosage forms and kits are also provided.

Description

[Technical field]

[0001] The present invention relates to the treatment of disorders affecting the eyelid muscles of a patient. [Background technology]

[0002] Disorders affecting the eyelid muscles can adversely affect the lives of patients who suffer from them. Among these disorders, blepharospasm and facial spasms (e.g. hemifacial spasm) are particularly unpleasant.

[0003] Blepharospasm is primarily characterized by abnormal contraction of the orbicularis oculi muscle. More specifically, blepharospasm manifests as uncontrollable excessive blinking and spasms of one or both eyes, and is further characterized by uncontrollable eyelid closure that lasts longer than the normal blink reflex. Symptoms of blepharospasm can be recurrent and may last for hours or days at a time, and in some cases, symptoms (such as spasms) can be chronic and persistent, causing lifelong difficulties for patients suffering from this health condition. Other symptoms may include spasms that spread to the nose, face, and neck, dry eyes, and sensitivity to the sun and bright light.

[0004] The cause of blepharospasm is unclear. It has been suggested that blepharospasm may be precipitated by certain medications, for example, medications used to treat Parkinson's disease, estrogen replacement therapy, and acute withdrawal of benzodiazepines. Blepharospasm may also be associated with brain disorders (including, for example, neurodegenerative diseases, dysfunction of the basal ganglia of the brain, and multiple sclerosis), brain injury, or head trauma (for example, concussion).

[0005] Hemifacial spasm is a movement disorder characterized by involuntary tonic-clonic contractions of the facial muscles on one side of the face. Bilateral cases are occasionally seen, but are extremely rare. The affected muscles are those innervated by the facial nerve (cranial nerve VII). The initial symptoms of the disorder usually appear in the orbicularis oculi muscle and may spread to other facial muscles. There are two types of hemifacial spasm (HFS): typical HFS and atypical HFS. In typical HFS, the spasms / twitches usually start in the orbicularis oculi muscle of the lower eyelid. Over time, they spread to the entire eyelid, then the orbicularis oris muscle around the lips and the buccinator muscle in the zygomatic region. In atypical HFS, the spasms usually start in the orbicularis oris muscle around the lips and the buccinator muscle in the zygomatic region of the lower face, then progress over time to the orbicularis oculi muscle of the eyelid. The typical form is the most common, while the atypical form is seen in only about 2-3% of people with hemifacial spasm.

[0006] Drug therapy for disorders affecting the eyelid muscles of patients has generally proven to be unpredictable and short-term. Anticholinergics, tranquilizers, and botulinum neurotoxins (e.g., Dysport®, Botox®, Xeomin®) are the most commonly used treatment options. However, these treatment options are suboptimal and are associated with serious side effects, including toxicity and unwanted paralysis of the facial muscles. In some cases, invasive surgical procedures may be considered for patients who do not respond adequately to drug therapy or botulinum neurotoxin injections. Thus, new and effective therapies for use in treating blepharospasm are constantly being tested or sought.

[0007] More specifically, Botulinum neurotoxin A (BoNT / A) selectively inhibits the release of acetylcholine from presynaptic nerve terminals, thereby inhibiting cholinergic transmission at the neuromuscular junction, inducing muscle contraction and decreased muscle tone, and relaxing the injected muscle. However, the duration of action of currently available BoNT / A products is approximately 12-14 weeks, during which time new nerve terminals sprout and restore normal nerve function, and the original symptoms recur. Therefore, to maintain efficacy, injections must be repeated periodically. Thus, the frequency of BoNT / A injections is an important consideration for the treatment of disorders affecting the patient's eyelid muscles (such as blepharospasm and / or hemifacial spasm), given the potential chronicity of the condition and the long-term nature of the required treatment. Indeed, this impacts on the direct and indirect health costs to the patient and caregiver, the logistics of injections within the hospital / clinic, and most importantly, the quality of life of the patient.

[0008] Dysport® is approved for the treatment of blepharospasm and hemifacial spasm, with a maximum total dose of 120 units per eye per treatment session. Clinicians must administer Dysport® to the patient's eyelid muscles up to a threshold upper limit of 120 units total per eye (240 units if both eyes are treated) per treatment session. Clinicians are forced to make difficult choices during the treatment of patients. In other words, in conventional treatment plans, clinicians must balance the relatively small total amount of BoNT / A that can be administered (necessary due to the high toxicity of BoNT / A) with the effective amount in multiple different muscles and / or their locations. As a result, certain muscles may be neglected while other muscles receive suboptimal amounts of BoNT / A, resulting in suboptimal treatment.

[0009] Furthermore, conventional treatment regimens for such disorders are complex and result in clinicians underdosing to avoid toxicity to the patient. Thus, there is a need for a convenient, safe and effective single dose unit and corresponding guidance regarding the number of units (including, for example, the number of injection sites per muscle) that can be administered to the eyelid muscle in a treatment session that does not result in toxicity to the patient.

[0010] In conclusion, there is a need for improved treatments for disorders affecting the eyelid muscles in patients (e.g., blepharospasm and / or hemifacial spasm) that allow for a personalized, patient-centered approach that avoids toxicity and provides long-lasting treatment (resulting in less frequent dosing), while tailoring treatment to the target clinical pattern, allowing for different combinations of muscles and / or sites of injection depending on the distribution, extent, and severity of the disorder.

[0011] The present invention overcomes one or more of the problems set forth above. Summary of the Invention

[0012] The inventors have surprisingly found that modified BoNT / A is particularly useful for treating disorders affecting the eyelid muscles of a patient, such as blepharospasm and / or hemifacial spasm. The modified BoNT / A comprises a BoNT / A light chain and translocation domain and a BoNT / B receptor binding domain (H C domain), resulting in a modified BoNT / A that exhibits increased retention at the site of administration (reduced diffusion away from the administration site) and / or extended duration of action (e.g., 6 to 9 months).

[0013] Advantageously, the modified BoNT / A has an improved safety profile when compared to unmodified BoNT / A (e.g., Dysport®), which may be expressed by the increased safety margins described herein for the modified BoNT / A.

[0014] Based on the preclinical and clinical data herein, it has been shown that a larger total dose of modified BoNT / A may be administered to a patient while achieving a similar safety profile to unmodified BoNT / A (e.g., Dysport®) at such higher doses. Thus, in treating a disorder affecting the eyelid muscles of a patient (e.g., blepharospasm and / or hemifacial spasm), more modified BoNT / A may be injected and / or more muscles and / or sites may be injected until a maximum total dose is reached. This is an important and advantageous discovery, improving the treatment of such diseases and providing clinicians with a wider range of treatment options. The treatment may be improved in that it provides a more durable treatment (resulting in less frequent administration) and / or can be tailored to the patient and / or results in an improved quality of life for the patient when compared to treatment with unmodified BoNT / A (e.g., Dysport®). Thus, the treatment of the present invention is improved compared to conventional treatment regimes.

[0015] Furthermore, the present invention provides a convenient, safe and effective single unit dose, as well as a total (maximum) dose that can be safely administered in a single treatment. The present invention also provides a guide to the number of times the unit dose can be administered into a muscle (including, for example, the number of injection sites per muscle) without resulting in toxicity to the patient. Thus, the treatment of a disorder affecting the eyelid muscles of a patient (e.g., blepharospasm and / or hemifacial spasm) according to the present invention is less complicated for the clinician, helping to avoid under-dosing and / or over-dosing. Furthermore, the treatment according to the present invention is much more satisfactory for the patient, since it is better adapted to the needs of the patient, when compared to conventional treatments. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] A broad aspect of the present invention provides: - a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient, the modified BoNT / A being administered by intramuscular injection to multiple sites on the patient's face; - a method of treating blepharospasm in a patient, the method comprising administering a modified BoNT / A by intramuscular injection to multiple sites on the patient's face; - use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating blepharospasm in a patient, the modified BoNT / A being administered by intramuscular injection to multiple sites on the patient's face; wherein the modified BoNT / A is administered as a unit dose containing at least 240 pg (preferably between 240 pg and 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A is a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0017] A further broad aspect of the invention provides: - a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating typical hemifacial spasm, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face; - a method of treating typical hemifacial spasm, comprising administering a modified BoNT / A by intramuscular injection to multiple sites on a patient's face; - use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for use in treating typical hemifacial spasm, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face; The modified BoNT / A is administered as a unit dose containing at least 240 pg (preferably between 240 pg and 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A is a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0018] A further broad aspect of the invention provides: - a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating atypical hemifacial spasm, said modified BoNT / A being administered by intramuscular injection to multiple sites on a patient's face; - a method of treating atypical hemifacial spasm, comprising administering a modified BoNT / A by intramuscular injection to multiple sites on a patient's face; - use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for use in the treatment of atypical hemifacial spasm, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face; wherein the modified BoNT / A is administered as a unit dose containing at least 240 pg (preferably between 240 pg and 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A is a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0019] One aspect provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient, said modified BoNT / A being administered by intramuscular injection to multiple sites on the patient's face, said method comprising: a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye of a patient, wherein said up to six different injection sites are selected from the following: i) the medial orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the medial orbicularis oculi superior muscle, preferably the anterior orbital septum); ii) superior orbital orbicularis oculi; iii) the lateral orbicularis oculi superior muscle (e.g. the anterior orbital septum or orbital portion of said lateral orbicularis oculi superior muscle, preferably the anterior orbital septum); iv) lateral orbital orbicularis oculi; v) medial superior tarsal preorbital orbicularis oculi; and vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or b) administering a unit dose per injection site of the modified BoNT / A to up to two different injection sites in the inferior orbicularis oculi muscle proximal to the first eye of the patient, wherein said up to two different injection sites are selected from the following: i) the medial orbicularis oculi inferior muscle (e.g., the preorbital septum or orbital portion of the medial orbicularis oculi inferior muscle, preferably the orbital septum portion); and ii) the lateral inferior orbicularis oculi muscle (e.g., the orbital septum or orbital part of the lateral inferior orbicularis oculi muscle, preferably the orbital septum); and / or c) administering a unit dose of the modified BoNT / A per injection site into up to two different injection sites selected from the following: i) two different injection sites in the corrugator supercilii muscle proximal to the patient's first eye; and ii) One site on the proximal canthus muscle proximal to the patient's first eye; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and the modified BoNT / A comprises a BoNT / A light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0020] One aspect provides a method of treating blepharospasm in a patient, the method comprising administering a modified botulinum neurotoxin A (BoNT / A) by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye of a patient, wherein said up to six different injection sites are selected from the following: i) the medial orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the medial orbicularis oculi superior muscle, preferably the anterior orbital septum); ii) superior orbital orbicularis oculi; iii) the lateral orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the lateral orbicularis oculi superior muscle, preferably the anterior orbital septum); iv) lateral orbital orbicularis oculi; v) medial superior anterior orbital orbicularis oculi; and vi) lateral upper eyelid anterior orbit orbicularis oculi; and / or b) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites in the inferior orbicularis oculi muscle proximal to the first eye of the patient, wherein said up to two different injection sites are selected from the following: i) the medial orbicularis oculi inferior muscle (e.g., the anterior orbital septum or orbital part of the medial orbicularis oculi inferior muscle, preferably the anterior orbital septum); and ii) the lateral inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of the lateral inferior orbicularis oculi muscle, preferably the anterior orbital septum); and / or c) administering a unit dose of the modified BoNT / A per injection site into up to two different injection sites selected from the following: i) two different injection sites in the corrugator supercilii muscle proximal to the patient's first eye; and ii) one site on the proximal canthus muscle of the patient's first eye; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and the modified BoNT / A comprises a BoNT / A light chain and a translocation domain (HN ) and the BoNT / B receptor binding domain (H C domain), and

[0021] One aspect provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating typical hemifacial spasm, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, the treatment comprising: (a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye of a patient, wherein said up to six different injection sites are selected from the following: (i) the medial orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the medial orbicularis oculi superior muscle, preferably the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital portion of the lateral orbicularis oculi superior muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or (b) administering a unit dose of the modified BoNT / A per injection site to up to two distinct injection sites in the inferior orbicularis oculi muscle proximal to the first eye of the patient, wherein said up to two distinct injection sites are selected from the following: (i) the medial orbicularis oculi inferior muscle (e.g., the anterior orbital septum or orbital portion of the medial orbicularis oculi inferior muscle, preferably the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of the lateral inferior orbicularis oculi muscle, preferably the anterior orbital septum); and / or (c) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii muscle proximal to the patient's first eye; and (ii) One site on the proximal canthus muscle of the patient's first eye; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and the modified BoNT / A comprises a BoNT / A light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0022] Throughout this disclosure, the injection site "lateral orbital orbicularis oculi" is referred to in the context of the "superiorbital orbicularis oculi" injection site, for example, because the "lateral orbital orbicularis oculi" site is closer to the upper eyelid than to the lower eyelid. That said, in any aspect of the embodiments described herein (whether for the treatment of blepharospasm, typical hemifacial spasm, or atypical hemifacial spasm), the following terms recited in (1) may be used synonymously with the following terms recited in (2): (1) administering a unit dose of modified BoNT / A per injection site to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye (and / or a second eye) of a patient, wherein the six different injection sites are selected from the following: (iii) the medial orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital portion of the medial orbicularis oculi superior muscle, preferably the anterior orbital septum); (iv) superior orbital orbicularis oculi; (v) the lateral superior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of the lateral superior orbicularis oculi muscle, preferably the anterior orbital septum); (vi) lateral orbitoorbicularis; (vii) medial superior tarsal plate anterior orbitoorbicularis oculi; (viii) lateral superior tarsal plate anterior orbitoorbicularis oculi muscle; (2) "Administering a unit dose of modified BoNT / A per injection site to up to six different injection sites in the superior and / or lateral orbital orbicularis oculi muscles proximal to a first (and / or second) eye of a patient, wherein said up to six different injection sites are selected from the following: (i) the medial orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the medial orbicularis oculi superior muscle, preferably the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the lateral orbicularis oculi superior muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) Lateral superior tarsal plate anterior orbital orbicularis oculi muscle.

[0023] The modified BoNT / A may be administered by intramuscular injection, preferably to at least six different sites on the patient's face. In other words, the modified BoNT / A may be administered by intramuscular injection, preferably to at least six different sites on the patient's face. In other words, the number of different injection sites at which a unit dose is administered may preferably be at least six.

[0024] The injection schedule of the present invention allows the clinician to provide a personalized pattern of muscle involvement in a patient's blepharospasm and direct where to inject into the exposed muscles while mitigating the risk of relieving ptosis.

[0025] If the pretarsal muscle of the upper eyelid is involved, up to two injections may be made, preferably in the pretarsal portion of the orbicularis oculi muscle of the upper eyelid. The plan preferably avoids the upper eyelid sulcus to reduce the chance of developing ptosis. If the corrugator supercilii / procerus muscles are involved, up to two injections are preferably made in the glabella on each side.

[0026] The method of treating blepharospasm may further include: (a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a second eye of a patient, wherein said up to six different injection sites are selected from the following: (i) the medial orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the medial orbicularis oculi superior muscle, preferably the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral orbicularis oculi superior muscle (e.g., the anterior orbital septum or orbital part of the lateral orbicularis oculi superior muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or (b) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites in the inferior orbicularis oculi muscle proximal to the second eye of the patient, wherein said up to two different injection sites are selected from the following: (i) the medial orbicularis oculi inferior muscle (e.g., the anterior orbital septum or orbital portion of the medial orbicularis oculi inferior muscle, preferably the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle; and / or (c) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii muscle proximal to the patient's second eye; and (ii) One site on the proximal canthus muscle of the patient's second eye.

[0027] The modified BoNT / A may be administered by intramuscular injection, preferably at at least six different sites on the patient's face. In other words, the modified BoNT / A may be administered by intramuscular injection, preferably at at least six different sites on the patient's face. In other words, the number of different injection sites at which unit doses are administered is preferably at least six.

[0028] The method for treating blepharospasm preferably includes: (a) administering a unit dose of the modified BoNT / A to the lateral superior orbicularis oculi muscle proximal to a first eye of the patient (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the lateral superior orbital septum anterior orbicularis oculi muscle); (b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle proximal to the first eye of the patient (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the medial anterior superior orbital septum orbicularis oculi muscle); and (c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle proximal to the first eye of the patient (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the lateral inferior orbicularis oculi).

[0029] In other words, a method for treating blepharospasm may preferably include administering a unit dose of modified BoNT / A to each of the following: (a) the lateral superior orbicularis oculi muscle proximal to the patient's first eye (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the lateral superior orbital septum anterior orbicularis oculi muscle); (b) the medial superior orbicularis oculi muscle proximal to the patient's first eye (e.g., the anterior orbital portion of the muscle, preferably the medial superior orbital portion of the anterior orbicularis oculi muscle); and (c) the lateral inferior orbicularis oculi muscle proximal to the patient's first eye (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the lateral inferior orbicularis oculi).

[0030] One aspect provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbital septum anterior orbicularis oculi muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dosage administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0031] In a related embodiment, the invention provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as the dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbital septum anterior orbicularis oculi muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0032] In a related embodiment, the invention provides a method of treating blepharospasm in a patient, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbital septum anterior orbicularis oculi muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0033] In a related embodiment, the invention provides a method of treating blepharospasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as the dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0034] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for use in treating blepharospasm in a patient, the medicament comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbital septum anterior orbicularis oculi muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dosage administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0035] In one aspect, the invention provides for the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating blepharospasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2 [such as the bichain form of SEQ ID NO:2]), comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbital septum anterior orbicularis oculi muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A contains a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0036] One embodiment provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating typical hemifacial spasm, said modified BoNT / A being administered by intramuscular injection to multiple sites on a patient's face, said method comprising: (a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye of a patient, wherein said up to six different injection sites are selected from the following: (i) the medial orbicularis oculi superior muscle (e.g. the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral superior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or (b) administering a unit dose of the modified BoNT / A per injection site to up to two distinct injection sites in the inferior orbicularis oculi muscle proximal to the first eye of the patient, wherein said up to two distinct injection sites are selected from the following: (i) the medial inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the anterior orbital septum); and / or (c) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii muscle proximal to the patient's first eye; and (ii) a site on the proximal canthus muscle of the patient's first eye; and / or (d) administering one or more unit doses of the modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of the modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of the modified BoNT / A, and the modified BoNT / A comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H domain). N ) and the BoNT / B receptor binding domain (H C domain), and

[0037] One embodiment provides a method of treating typical hemifacial spasm, the method comprising administering a modified BoNT / A by intramuscular injection to multiple sites on a patient's face, the method comprising: (a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye of a patient, wherein said up to six different injection sites are selected from the following: (i) the medial orbicularis oculi superior muscle (e.g. the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral superior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or (b) administering a unit dose of the modified BoNT / A per injection site to up to two distinct injection sites in the inferior orbicularis oculi muscle proximal to the first eye of the patient, wherein said up to two distinct injection sites are selected from the following: (i) the medial inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the anterior orbital septum); and / or (c) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii muscle proximal to the patient's first eye; and (ii) a site on the proximal canthus muscle of the patient's first eye; and / or (d) administering one or more unit doses of the modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and the modified BoNT / A comprises a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H domain). N ) and the BoNT / B receptor binding domain (H C domain), and

[0038] One aspect provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating typical hemifacial spasm, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, and said treatment includes: (a) administering a unit dose per injection site of a modified BoNT / A to up to six different injection sites in the orbicularis oculi superior muscle proximal to a first eye of a patient, wherein said up to six different injection sites are selected from the following: (i) the medial orbicularis oculi superior muscle (e.g. the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral superior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi muscle; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi muscle; and / or (b) administering a unit dose of the modified BoNT / A per injection site to up to two distinct injection sites in the inferior orbicularis oculi muscle proximal to the first eye of the patient, wherein said up to two distinct injection sites are selected from the following: (i) the medial inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the anterior orbital septum); and / or (c) administering a unit dose of the modified BoNT / A per injection site to up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii muscle proximal to the patient's first eye; and (ii) a site on the proximal canthus muscle of the patient's first eye; and / or (d) administering one or more unit doses of the modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, and the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A comprises a botulinum neurotoxin A (BoNT / A) light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0039] The modified BoNT / A may be administered by intramuscular injection, preferably at at least six different sites on the patient's face. In other words, the modified BoNT / A may be administered by intramuscular injection, preferably at at least six different sites on the patient's face. In other words, the number of different injection sites at which a unit dose is administered may preferably be at least six.

[0040] The method for treating typical hemifacial spasm preferably includes: a) administering a unit dose of the modified BoNT / A to the lateral superior orbicularis oculi muscle proximal to the first eye of the patient (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the lateral superior orbital septum anterior orbicularis oculi muscle); b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle proximal to the first eye of the patient (e.g., the preorbital or orbital portion of the muscle, preferably the medial superior orbicularis oculi muscle); and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle proximal to the first eye of the patient (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the lateral inferior orbital septum anterior orbicularis oculi muscle); In other words, a method for treating typical hemifacial spasm may preferably include administering a unit dose of modified BoNT / A to each of the following: a) the lateral superior orbicularis oculi muscle proximal to the patient's first eye (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the lateral superior orbital septum anterior orbicularis oculi muscle); b) the medial superior orbicularis oculi muscle proximal to the patient's first eye (e.g., the anterior orbital portion of the muscle, preferably the medial superior orbital septal anterior orbicularis oculi muscle); and c) the lateral inferior orbicularis oculi muscle proximal to the patient's first eye (e.g., the anterior orbital septum or orbital portion of said muscle, preferably the lateral inferior orbital septum anterior orbicularis oculi muscle).

[0041] Another embodiment provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating typical hemifacial spasm, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) adjacent to the eye affected by the hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the eye affected by the hemifacial spasm); c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior anterior orbital septum orbicularis oculi muscle) adjacent to the eye affected by the hemifacial spasm; and d) administering one or more unit doses of the modified BoNT / A to one or more additional muscles affected by hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A contains a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0042] In a related aspect, the invention provides a modified BoNT / A for use in a method of treating atypical hemifacial spasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as a two-chain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) adjacent to the eye affected by the hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the eye affected by the hemifacial spasm); c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle adjacent to the eye affected by hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A contains a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0043] In a related aspect, the invention provides a method of treating typical hemifacial spasm in a patient, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) adjacent to the eye affected by the hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the eye affected by the hemifacial spasm); c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior anterior orbital septum orbicularis oculi muscle) adjacent to the eye affected by the hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0044] In a related embodiment, the invention provides a method of treating typical hemifacial spasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as the dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) adjacent to the eye affected by the hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the eye affected by the hemifacial spasm); c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior anterior orbital septum orbicularis oculi muscle) adjacent to the eye affected by the hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0045] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating typical hemifacial spasm in a patient, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) adjacent to the eye affected by the hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the eye affected by the hemifacial spasm); c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior anterior orbital septum orbicularis oculi muscle) adjacent to the eye affected by the hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0046] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating typical hemifacial spasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as a dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) adjacent to the eye affected by the hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the eye affected by the hemifacial spasm); c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior anterior orbital septum orbicularis oculi muscle) adjacent to the eye affected by the hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0047] One embodiment provides a modified botulinum neurotoxin A (BoNT / A) for use in treating atypical hemifacial spasm, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm); b) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) 1 unit dose into the buccinator muscle; (vii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (viii) one unit dose into the nasal bridge; (ix) one unit dose into the levator palpebrae superioris; and / or c) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: Unit dose per injection site at up to six different injection sites in the orbicularis oculi superior muscle selected from: (i) the medial superior orbital septum preorbicularis oculi muscle (e.g., in the anterior orbital septum or orbital portion of the muscle, preferably located in the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) lateral superior orbital septum preorbicularis oculi (e.g., located in the anterior orbital septum or orbital part of the muscle, preferably in the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or Unit dose per injection site at up to two different injection sites in the inferior orbicularis oculi muscle selected from: (i) the medial orbicularis oculi inferior muscle (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); and / or Unit dose per injection site at up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii; and (ii) one site on the procerus muscle; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and N ) and the BoNT / B receptor binding domain (H C domain), and a modified BoNT / A comprising.

[0048] One embodiment provides a method of treating atypical hemifacial spasm, the method comprising administering a modified BoNT / A by intramuscular injection to multiple sites on a patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm); b) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (x) 1 unit dose into the zygomaticus major muscle; (xi) 1 unit dose into the zygomaticus minor muscle; (xii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (xiii) one unit dose into the mentalis muscle; (xiv) 1 unit dose into the platysma muscle; (xv) 1 unit dose into the buccinator muscle; (xvi) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xvii) one unit dose into the nasal bridge; (xviii) one unit dose into the levator palpebrae superioris; and / or c) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: Unit dose per injection site at up to six different injection sites in the orbicularis oculi superior muscle selected from: (i) the medial orbicularis oculi superior muscle (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) lateral superior orbicularis oculi (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or Unit dose per injection site at up to two different injection sites in the inferior orbicularis oculi muscle selected from: (iii) the medial orbicularis oculi inferior muscle (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); and (iv) the lateral inferior orbicularis oculi muscle (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); and / or Unit dose per injection site at up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii; and (ii) one site on the procerus muscle; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and N ) and the BoNT / B receptor binding domain (H C domain), and a modified BoNT / A comprising.

[0049] One embodiment provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating atypical hemifacial spasm, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, and said treatment comprises: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm); b) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) 1 unit dose into the buccinator muscle; (vii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (viii) one unit dose into the nasal bridge; (ix) one unit dose into the levator palpebrae superioris; and / or c) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: Unit dose per injection site at up to six different injection sites in the orbicularis oculi superior muscle selected from: (i) the medial orbicularis oculi superior muscle (e.g., located in the anterior orbital septum or orbital portion of the muscle, preferably in the anterior orbital septum); (ii) superior orbital orbicularis oculi; (iii) the lateral superior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of said muscle, preferably the anterior orbital septum); (iv) lateral orbital orbicularis oculi; (v) medial superior tarsal preorbital orbicularis oculi; and (vi) lateral superior tarsal plate anterior orbital orbicularis oculi; and / or Unit dose per injection site at up to two different injection sites in the inferior orbicularis oculi muscle selected from: (i) the medial inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital portion of the muscle, preferably the anterior orbital septum); and (ii) the lateral inferior orbicularis oculi muscle (e.g., the anterior orbital septum or orbital part of the muscle, preferably the anterior orbital septum); and / or Unit dose per injection site at up to two different injection sites selected from the following: (i) two different injection sites in the corrugator supercilii; and (ii) one site on the procerus muscle; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and the modified BoNT / A is a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H domain). N ) and the BoNT / B receptor binding domain (H C domain), and

[0050] Another embodiment provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating atypical hemifacial spasm, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris muscle (e.g., administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and / or inferior muscles; preferably, administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and inferior muscles); b) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and a translocation domain (H N ) and the BoNT / B receptor binding domain (H C domain), and

[0051] In a related aspect, the invention provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as the dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris muscle (e.g., administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and / or inferior muscles; preferably, administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and inferior muscles); and b) optionally administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0052] In a related embodiment, the invention provides a method of treating atypical hemifacial spasm in a patient, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris muscle (e.g., administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and / or inferior muscles; preferably, administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and inferior muscles); and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0053] In a related embodiment, the invention provides a method of treating atypical hemifacial spasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as a bichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris muscle (e.g., administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and / or inferior muscles; preferably, administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and inferior muscles); and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0054] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for the treatment of atypical hemifacial spasm in a patient, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris muscle (e.g., administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and / or inferior muscles; preferably, administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and inferior muscles); and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0055] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for the treatment of atypical hemifacial spasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as a bichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris muscle (e.g., administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and / or inferior muscles; preferably, administering one unit dose of modified BoNT / A to the hemifacial spasm-affected orbicularis oris superior and inferior muscles); and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of modified BoNT / A is at least 240 pg (preferably 240 pg to 8,000 pg) of modified BoNT / A; The total dose administered during treatment is greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) and up to 82,500 pg (e.g., up to 75,000 pg) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain (HN) and a BoNT / B receptor binding domain (HC domain).

[0056] Throughout this disclosure, references to a total dose of modified BoNT / A administered during treatment of greater than 24,100 pg may mean that the total dose administered during treatment is greater than 24,100 pg of modified BoNT / A. More preferably, references to a total dose of modified BoNT / A administered during treatment of greater than 24,000 pg may mean that the total dose administered during treatment is greater than 35,000 pg of modified BoNT / A.

[0057] The term "typical hemifacial spasm" may be used synonymously with the term "hemifacial spasm" throughout this disclosure.

[0058] The term "treating blepharospasm in a patient for a longer duration than when treated with unmodified BoNT / A" may mean that after administration of modified BoNT / A, one or more symptoms of the disorder in the patient are reduced for a longer period (e.g., 6 to 9 months) compared to administration of unmodified BoNT / A. The reduction may be determined by comparison with a comparable control subject exhibiting a comparable symptom treated with unmodified BoNT / A. During a period during which the severity of one or more symptoms of the control subject is substantially the same (e.g., the same) as before unmodified BoNT / A treatment, the patient treated with modified BoNT / A according to the present invention may show at least a 5%, 10%, 25%, or 50% improvement in the comparable one or more symptoms compared to the severity of the one or more symptoms before treatment with the modified BoNT / A. The unmodified BoNT / A is preferably SEQ ID NO: 2, which exists in a two-chain form.

[0059] The term "treating a patient's typical hemifacial spasm for a longer duration than when treated with unmodified BoNT / A" may mean that one or more symptoms of the disorder in a patient are reduced for a longer period (e.g., 6 to 9 months) after administration of unmodified BoNT / A compared to administration of unmodified BoNT / A. The reduction may be determined by comparison with a comparable control subject exhibiting comparable symptoms treated with unmodified BoNT / A. During a period in which the severity of one or more symptoms in the control subject is substantially the same (e.g., the same) as before unmodified BoNT / A treatment, a patient treated with a modified BoNT / A according to the present invention may show at least a 5%, 10%, 25%, or 50% improvement in the comparable one or more symptoms compared to the severity of the one or more symptoms before treatment with the modified BoNT / A.

[0060] The term "treating a patient with atypical hemifacial spasm for a longer duration than those treated with unmodified BoNT / A" may mean that one or more symptoms of the disorder in a patient are reduced for a longer period (e.g., 6 to 9 months) after administration of a modified BoNT / A of the present invention compared to administration of unmodified BoNT / A. The reduction may be determined by comparison with comparable control subjects exhibiting comparable symptoms treated with unmodified BoNT / A. During a period in which the severity of one or more symptoms in the control subject is substantially the same (e.g., the same) as before unmodified BoNT / A treatment, a patient treated with a modified BoNT / A according to the present invention may show at least a 5%, 10%, 25%, or 50% improvement in the comparable one or more symptoms compared to the severity of the one or more symptoms before treatment with the modified BoNT / A.

[0061] The unit dose may be at least 240.4 pg, 500 pg, 1,000 pg, 2,000 pg, 3,000 pg or 4,000 pg, for example at least 1,000 pg of modified BoNT / A.

[0062] The unit dose may be at least 240.4 pg, 500 pg, 1,000 pg, 2,000 pg, 3,000 pg, 4,000 pg, 5,000 pg, 6000 pg, or 7,000 pg, preferably at least 4,000 pg of the modified BoNT / A.

[0063] In one embodiment, the upper limit of the unit dose of the present invention can be determined based on the total dose administered during treatment and the number of muscles and / or sites to which modified BoNT / A is administered. For example, if the total dose administered during treatment of blepharospasm is up to 75,000 pg of modified BoNT / A and the administration includes (i) a (single) unit dose to the lateral superior orbicularis oculi muscle proximal to the first eye of the patient, (ii) a (single) unit dose to the medial superior orbicularis oculi muscle proximal to the first eye, (iii) a (single) unit dose only to the lateral inferior orbicularis oculi muscle proximal to the first eye, (iv) two unit doses only to the frontalis muscle proximal to the first eye, (v) two unit doses only to the corrugator supercilii muscle proximal to the first eye, and (vi) a (single) unit dose to the procerus muscle (e.g., 8 unit doses to multiple sites), the upper limit of the unit dose can be 9,375 pg. The upper limit may be 5,000 pg if the booster doses include (i) a (single) unit dose to the lateral orbicularis oculi superior muscle proximal to the patient's second eye, (ii) a (single) unit dose to the medial orbicularis oculi superior muscle proximal to the second eye, (iii) a (single) unit dose to the lateral orbicularis oculi inferior muscle proximal to the second eye, (iv) two unit doses to the frontalis muscle proximal to the second eye, and (v) two unit doses to the corrugator supercilii muscle proximal to the second eye (e.g., an additional 7 unit doses, bringing the total number of unit doses to 15).

[0064] The unit dose may be 240 pg to 8,000 pg of modified BoNT / A, wherein the modified BoNT / A comprises a BoNT / A light chain and translocation domain and a BoNT / B receptor binding domain (H CThe upper end of the unit dose range may be 7,500, 7,000, 6,500, 6,000, 5,500, 5,000, 4,800, 4,500, 4,000, 3,500, 3,000, 2,500, 2,400, 2,000, 1,500, or 1,250 pg of modified BoNT / A. The upper end of the unit dose range may be 5,500, 5,000, or 4,800 pg of modified BoNT / A. A preferred upper end of the unit dose range may be 5,000 pg of modified BoNT / A. The lower limit of the unit dose range may be 300, 400, 500, 600, 700, 800, 900, 1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, or 5,000 pg of modified BoNT / A, preferably the lower limit is 1,000 pg. The unit dose may be 1,000 pg to 6,000 pg, 1,000 pg to 5,500 pg, 1,000 pg to 5,000 pg, or 1,000 pg to 4,500 pg of modified BoNT / A. The unit dose may be between 1,000 pg and 4,800 pg, between 1,000 pg and 4,000 pg, between 1,000 pg and 2,400 pg, or between 1,000 pg and 2,000 pg.

[0065] The unit dose may be 1,500 to 5,000 pg of modified BoNT / A, preferably 2,000 pg to 4,500 pg of modified BoNT / A. Examples of suitable unit doses include about 2,500 pg (e.g., 2,000 pg ± 10%) of modified BoNT / A: and about 4,000 pg (e.g., 4,000 pg ± 10%) of modified BoNT / A. Such unit doses are particularly suitable for treating blepharospasm (bilateral or unilateral).

[0066] The total dose administered in practicing the treatment regimen of the present invention may be greater than 24,000 pg (e.g., preferably greater than 24,100 pg; more preferably greater than 35,000 pg) of modified BoNT / A. In other words, the total amount of modified BoNT / A administered in a given treatment session may be greater than 24,000 pg. The total dose may be up to 82,500, 80,000, 75,000, 70,000, 65,000, 60,000, 55,000, 50,000, 45,000, 40,000, 35,000, 30,000, or 25,000 pg. The total dose may be at least 24,500, 25,000, 30,000, 35,000, 40,000, 45,000, 50,000, 55,000, 60,000, 65,000 or 70,000 pg. Preferably, the total dose is at least 30,000 pg of modified BoNT / A. The total dose may be greater than 24,000 pg and up to 70,000 pg, preferably between 30,000 pg and 60,000 pg.

[0067] The total dose may be greater than 35,000 pg and up to 70,000 pg.

[0068] The total dose may be between 25,000 pg and 50,000 pg (e.g., 25,000 pg to 50,000 pg) of modified BoNT / A. Examples of suitable ranges of total doses include 25,000 pg to 35,000 pg of modified BoNT / A; and 45,000 pg to 50,000 pg of modified BoNT / A. Examples of suitable total doses include about 30,000 pg (e.g., 30,000 pg ± 10%) and about 48,000 pg (e.g., 48,000 pg ± 10%) of modified BoNT / A. Such unit doses are particularly suitable for treating blepharospasm (bilateral or unilateral).

[0069] Thus, a unit dose may be at least 240 pg of modified BoNT / A, and the total dose administered in practicing the therapeutic regimen of the invention may be greater than 24,000 pg and up to 82,500 pg of modified BoNT / A. A unit dose may be at least 240 pg of modified BoNT / A, and the total dose administered in practicing the therapeutic regimen of the invention may be greater than 35,000 pg and up to 82,500 pg of modified BoNT / A.

[0070] In a preferred embodiment, the unit dose may be at least 240 pg of modified BoNT / A and the total dose administered in practicing the therapeutic regimen of the invention may be greater than 24,000 pg and up to 75,000 pg of modified BoNT / A. The unit dose may be at least 240 pg of modified BoNT / A and the total dose administered in practicing the therapeutic regimen of the invention may be greater than 35,000 pg and up to 75,000 pg of modified BoNT / A.

[0071] In another preferred embodiment, the unit dose may be up to 5,000 pg of modified BoNT / A and the total dose administered in practicing the therapeutic regimen of the present invention may be greater than 24,000 pg and up to 75,000 pg of modified BoNT / A. The unit dose may be up to 5,000 pg of modified BoNT / A and the total dose administered in practicing the therapeutic regimen of the present invention may be greater than 35,000 pg and up to 75,000 pg of modified BoNT / A.

[0072] In another preferred embodiment, the unit dose may be 2,000 to 4,500 pg of modified BoNT / A, and the total dose administered in practicing the treatment regimen of the present invention is 25,000 pg to 50,000 pg of modified BoNT / A. Such regimens may be particularly suitable for treating blepharospasm (bilateral or unilateral). For example, suitable dosing regimens include: - a unit dose of modified BoNT / A of about 2,500 pg (e.g., 2,000 pg ± 10%) and a total dose of modified BoNT / A of about 30,000 pg (e.g., 30,000 pg ± 10%) - A unit dose of modified BoNT / A of about 4,000 pg (e.g., 4,000 pg ± 10%) and a total dose of modified BoNT / A of about 48,000 pg (e.g., 48,000 pg ± 10%).

[0073] In the case of unilateral disease (e.g., affecting one side of the face, such as one eye in unilateral blepharospasm), the total dose may refer to the total dose administered to the affected side of the face. In the case of bilateral disease (e.g., affecting both sides of the face, such as both eyes in unilateral blepharospasm), the total dose may refer to the total dose administered to both sides of the face. It is preferred that 50% of the total dose is administered to each side of the face (e.g., each eye in the case of bilateral blepharospasm).

[0074] One aspect provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient, said modified BoNT / A being administered by intramuscular injection to multiple sites on the patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ), the total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and the modified BoNT / A comprises a BoNT / A light chain and translocation domain and a BoNT / B receptor binding domain (H C domain), and

[0075] In a related aspect, the invention provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating blepharospasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as the dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to wherein the total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A; and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0076] In a related embodiment, the invention provides a method of treating blepharospasm in a patient, wherein a modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the proximal lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) of the patient's first eye; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 U is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0077] In a related embodiment, the invention provides a method of treating blepharospasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as bichain SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 U is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0078] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for use in treating blepharospasm in a patient, comprising a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 U is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0079] In one aspect, the invention provides for the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating blepharospasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2 [SEQ ID NO:2 for the bichain form]), comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior muscle) proximal to a first eye of a patient; b) administering a unit dose of the modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior muscle) proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the medial superior orbital septum anterior orbicularis oculi muscle) proximal to the first eye of the patient; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0080] Another aspect provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating typical hemifacial spasm, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by the hemifacial spasm; c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0081] In a related embodiment, the invention provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating typical hemifacial spasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as the dichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior pretarsal orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by the hemifacial spasm; c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior pretarsal orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; and d) administering one or more unit doses of the modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0082] In a related aspect, the invention provides a method of treating typical hemifacial spasm in a patient, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by the hemifacial spasm; c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; and d) administering one or more unit doses of the modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0083] In a related embodiment, the invention provides a method of treating typical hemifacial spasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2 [two-chain form of SEQ ID NO:2]), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by the hemifacial spasm; c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; and d) administering one or more unit doses of the modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0084] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating typical hemifacial spasm in a patient, the modified BoNT / A being administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by the hemifacial spasm; c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0085] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating typical hemifacial spasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2 [SEQ ID NO:2 for the dichain form]), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the lateral superior orbicularis oculi muscle (preferably the lateral superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by hemifacial spasm; b) administering a unit dose of modified BoNT / A to the medial superior orbicularis oculi muscle (preferably the medial superior orbicularis oculi anterior to the orbital septum) proximal to the eye affected by the hemifacial spasm; c) administering a unit dose of modified BoNT / A to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) proximal to the eye affected by hemifacial spasm; and d) administering one or more unit doses of modified BoNT / A to said hemifacial spasm-affected one or more additional muscles according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0086] Another aspect provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating atypical hemifacial spasm, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on a patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior orbicularis oris muscle affected by hemifacial spasm and / or administering one unit dose of modified BoNT / A to the inferior orbicularis oris muscle affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the orbicul; and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0087] In a related embodiment, the invention provides a modified botulinum neurotoxin A (BoNT / A) for use in a method of treating atypical hemifacial spasm in a patient for a longer duration than when treated with unmodified BoNT / A (e.g., SEQ ID NO:2 [dual-chain SEQ ID NO:2]), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm; and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0088] In a related embodiment, the invention provides a method of treating atypical hemifacial spasm in a patient, wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm; and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0089] In a related embodiment, the invention provides a method of treating atypical hemifacial spasm in a patient for a longer duration than when treated with an unmodified BoNT / A (e.g., SEQ ID NO:2 [two-chain form of SEQ ID NO:2]), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, the method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm; and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0090] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating atypical hemifacial spasm in a patient, wherein said modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, said method comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm; and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0091] In one aspect, the invention provides the use of a modified botulinum neurotoxin A (BoNT / A) in the manufacture of a medicament for treating atypical hemifacial spasm in a patient for a longer duration than treatment with an unmodified BoNT / A (e.g., SEQ ID NO:2, such as a bichain form of SEQ ID NO:2), wherein the modified BoNT / A is administered by intramuscular injection to multiple sites on the patient's face, comprising: a) administering a unit dose of modified BoNT / A to the orbicularis oris muscle affected by hemifacial spasm (e.g., administering one unit dose of modified BoNT / A to the superior and / or inferior orbicularis oris muscles affected by hemifacial spasm; preferably, administering one unit dose of modified BoNT / A to the superior and inferior orbicularis oris muscles affected by said hemifacial spasm; and b) optionally administering one or more unit doses of said modified BoNT / A to one or more additional muscles affected by said hemifacial spasm according to the following dosing schedule: (i) 1 unit dose into the zygomatic major muscle; (ii) 1 unit dose into the zygomaticus major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (iv) 1 unit dose into the mentalis muscle; (v) 1 unit dose into the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (vii) 1 unit dose into the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (ix) 1 unit dose into the procerus muscle; (x) 1 unit dose into the nasal bridge; (xi) 1 unit dose into the lateral superior orbicularis oculi muscle; (xii) 1 unit dose into the medial superior orbicularis oculi muscle; (xiii) one unit dose into the lateral inferior orbicularis oculi muscle; and / or (xiv) one unit dose into the levator palpebrae superioris; wherein the unit dose of the modified BoNT / A is at least 10 units (U) of modified BoNT / A, where 1 unit is the calculated median lethal dose (LD ) in mice. 50 ) is the amount of modified BoNT / A equivalent to The total dose administered during treatment is greater than 998 U and up to 3,432 U (e.g., up to 3,120 U) of modified BoNT / A, and The modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0092] The unit dose may be at least 21 U, 42 U, 83 U, 125 U, or 166 U, preferably at least 42 U, wherein the modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain).

[0093] The unit dose may be 10 U to 332.7 U of modified BoNT / A, which comprises a BoNT / A light chain and translocation domain and a BoNT / B receptor binding domain (H CThe upper limit of the unit dose range may be 312, 291, 270, 250, 229, 208, 199.6, 187, 166.3, 146, 125, 104, 99.8, 89.17, 62, or 52 U of modified BoNT / A. The lower limit of the unit dose range may be 12, 17, 21, 25, 29, 33, 37, 42, 62, 83, 104, 125, 146, 166, 187, or 208 U of modified BoNT / A, preferably the lower limit is 42 U. The unit dose may be 42 U to 199.6 U, 42 U to 166.3 U, 42 U to 99.8 U, or 42 U to 83.17 U.

[0094] The unit dose may be between 62.4 and 208 U of the modified BoNT / A, preferably between 83.2 and 187.2 U of the modified BoNT / A. Examples of suitable unit doses include about 104 U (e.g., 104 U ± 10%); and about 166.4 U (e.g., 166.4 U ± 10%) of the modified BoNT / A. Such unit doses are particularly suitable for treating blepharospasm (bilateral or unilateral).

[0095] The total dose administered in carrying out the treatment regimen of the present invention may be more than 998 U of modified BoNT / A. In other words, the total amount of modified BoNT / A administered in a given treatment session may be more than 998 U. The total dose may be up to 3430 U, 3326 U, 3119 U, 2911 U, 2703 U, 2495 U, 2287 U, 2079 U, 1871 U, 1663 U, 1455 U, 1247 U, or 1040 U. The total dose may be at least 1019 U, 1040 U, ​​1247 U, 1455 U, 1663 U, 1871 U, 2079 U, 2287 U, 2495 U, 2703 U, or 2911 U. Preferably, the total dose may be at least 125 U of modified BoNT / A. The total dose may be greater than 998U up to 2911U, preferably 1247U to 2495U.

[0096] The total dose may be between 1,040 U and 2080 U of modified BoNT / A. Examples of suitable ranges of total doses include 1,040 U to 1456 U of modified BoNT / A; and 1871.9 U to 2079.9 U of modified BoNT / A. Examples of suitable total doses include about 1247.9 U (e.g., 1247.9 U ± 10%) of modified BoNT / A; and about 1996.7 U (e.g., 1996.7 U ± 10%) of modified BoNT / A. Such unit doses are particularly suitable for treating blepharospasm (bilateral or unilateral).

[0097] Thus, the unit dose may be at least 10 U of modified BoNT / A, and the total dose administered in practicing the therapeutic regimen of the present invention may be greater than 998 U and up to 3430 U of modified BoNT / A. In a preferred embodiment, the unit dose may be at least 10 U of modified BoNT / A, and the total dose administered in practicing the therapeutic regimen of the present invention may be greater than 998 U and up to 3119 U of modified BoNT / A. In another preferred embodiment, the unit dose may be up to 208 U of modified BoNT / A, and the total dose administered in practicing the therapeutic regimen of the present invention may be greater than 998 U and up to 3119 U of modified BoNT / A.

[0098] In another preferred embodiment, the unit dose may be between 83.2 U and 187.2 U of modified BoNT / A, and the total dose administered in practicing the treatment regimen of the present invention may be between 1039.9 U and 2079.9 U of modified BoNT / A. Such regimens may be particularly suitable for treating blepharospasm (bilateral or unilateral). For example, suitable dosing regimens include: - a unit dose of about 104U (e.g., 104U ± 10%) of modified BoNT / A, and a total dose of about 1247.9U (e.g., 1247.9 units ± 10%) of modified BoNT / A - a unit dose of about 166.4 U (e.g., 166.4 U ± 10%) of modified BoNT / A, and a total dose of about 1996.7 U (e.g., 1996.7 U ± 10%) of modified BoNT / A.

[0099] In the case of unilateral disease (e.g., affecting one side of the face, such as one eye in unilateral blepharospasm), the total dose may refer to the total dose administered to the affected side of the face. In the case of bilateral disease (e.g., affecting both sides of the face, such as both eyes in unilateral blepharospasm), the total dose may refer to the total dose administered to both sides of the face. It is preferred that 50% of the total dose is administered to both sides of the face (e.g., each eye in the case of bilateral blepharospasm).

[0100] Suitable disorders affecting the eyelid muscles (e.g., affecting two or more eyelid muscles) include blepharospasm and facial spasm (e.g., hemifacial spasm). Preferably, the disorder affecting the patient's eyelid muscles is blepharospasm. Thus, the present invention may be directed to the treatment of blepharospasm and / or facial spasm (e.g., hemifacial spasm), and is preferably directed to the treatment of blepharospasm.

[0101] The present invention is anticipated based on the surprising results of dose escalation studies demonstrating that high dose therapy of Clostridial neurotoxins (e.g., greater than 24,000 pg and up to 82,500 pg) can be used to treat these disorders without exceeding acceptable toxicity levels (see Examples).

[0102] The disorder affecting the patient's eyelid muscles may be an eyelid muscle disorder. The cause of the disorder may be a nerve-related disorder (e.g., seventh nerve disorder).

[0103] The modified BoNT / A may be administered to any muscle affected by the disease (e.g., an affected eyelid muscle) that may contribute to (e.g., cause) one or more symptoms of the disease (e.g., blepharospasm and / or facial spasms, such as hemifacial spasm).

[0104] In one embodiment (preferably an embodiment directed to the treatment of blepharospasm), only a single unit dose of modified BoNT / A is administered at least to each of the following: the lateral superior orbicularis oculi muscle proximal to the patient's first eye (e.g., the lateral pretarsal orbicularis oculi muscle of the upper eyelid); the medial superior orbicularis oculi muscle proximal to the patient's first eye (e.g., the medial pretarsal orbicularis oculi muscle of the upper eyelid); and the lateral inferior orbicularis oculi muscle proximal to the patient's first eye (e.g., the lateral pretarsal orbicularis oculi muscle of the lower eyelid). Preferably, a single dose is administered per injection site, which in this embodiment may correspond to administration at three injection sites. Thus, although three unit doses may be administered in accordance with the above, the total number of unit doses administered may be greater than three, since additional muscles and / or sites may be treated in accordance with the present invention.

[0105] In one embodiment (preferably an embodiment directed to the treatment of blepharospasm), where the disease affects the eyelid muscles proximal to both eyes of the patient (e.g., bilateral blepharospasm), only a single unit dose of modified BoNT / A may be administered to each of the following: the lateral superior orbicularis oculi muscle proximal to the first eye of the patient; the medial superior orbicularis oculi muscle proximal to the first eye of the patient; the lateral inferior orbicularis oculi muscle proximal to the first eye of the patient; the lateral superior orbicularis oculi muscle proximal to the second eye of the patient; the medial superior orbicularis oculi muscle proximal to the second eye of the patient; and the lateral inferior orbicularis oculi muscle proximal to the second eye of the patient. Preferably, a single unit is administered per injection site, which in this embodiment corresponds to administration to six injection sites. Thus, six unit doses may be administered as described above, but the total number of unit doses administered may be greater than six, since additional muscles and / or sites may be treated according to the present invention.

[0106] The terms "first eye" and "second eye" may refer to either the left eye or the right eye. The terms are used only to distinguish the two eyes from each other. In other words, if the first eye is the left eye, the second eye is the right eye, and vice versa. Reference to the "first eye" does not imply that the muscles and / or areas proximal to the "second eye" always need to be treated. For example, the "first eye" may be referred to in the context of a unilateral disorder, such as unilateral blepharospasm, in which case the muscles and / or areas proximal to the second eye are not affected and therefore are not treated.

[0107] The term "proximal" means that the muscle and / or portion thereof being referenced is closest to the eye being referenced. For example, if a first eye is a patient's left eye, then a muscle and / or portion thereof that is proximal to the first eye is a muscle and / or portion thereof that is closer to the patient's left eye than to the patient's right eye.

[0108] The modified BoNT / A may be administered to one or more additional muscles and / or sites thereof. When administered to additional muscles and / or sites thereof, it is preferred to set an upper unit dose limit such that the total amount of modified BoNT / A administered does not exceed the total dose administered during treatment as defined in accordance with the present invention.

[0109] The additional muscles and / or regions thereof to be treated may be one or more (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, or all muscles and / or regions) selected from the following: medial inferior orbicularis oculi, orbicularis oris (e.g., superior and / or inferior orbicularis oris); zygomaticus (e.g., zygomaticus major); nasal muscles; mentalis; platysma; frontalis; corrugator supercilii; buccinator; masseter; procerus; and lateral canthus. The additional muscles and / or regions thereof to be treated may be one or more (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, or all of the muscles and / or regions) selected from the following: medial orbicularis oculi inferior, orbicularis oris superior, orbicularis oris inferior, zygomaticus major, zygomaticus minor, frontalis, mentalis, platysma, corrugator supercilii, buccinator, masseter, procerus, nasalis, and levator palpebrae superioris. The additional muscles and / or regions thereof to be treated may be one or more (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, or all of the muscles and / or regions) selected from the following: orbicularis oris superior, orbicularis oris inferior, zygomaticus major, zygomaticus minor, frontalis, mentalis, platysma, corrugator supercilii, buccinator, masseter, procerus, nasalis, and levator palpebrae superioris.

[0110] Where a muscle is recited for administration in accordance with the treatment of the invention, the invention may further include administering additional muscles not recited.

[0111] If desired, muscles and / or areas proximal to one or both eyes may be treated. At least a single unit dose may be administered to said muscles and / or areas, for example, two or more (e.g., three or more, four or more, or five or more) unit doses may be administered.

[0112] The modified BoNT / A may also be administered to the medial inferior orbicularis oculi muscle (e.g., the medial superior orbital septum preorbicularis oculi muscle of the lower eyelid). In one embodiment, only a single unit dose may be administered to the medial inferior orbicularis oculi muscle. If desired, the proximal medial inferior orbicularis oculi muscle of one or both eyes may be treated.

[0113] The modified BoNT / A may also be administered to the frontalis muscle. In one embodiment, at least a single unit dose may be administered to the frontalis muscle, for example, two or more, three or more, four or more, or five or more unit doses may be administered. If desired, the frontalis muscle proximal to one or both eyes may be treated.

[0114] The modified BoNT / A may also be administered to the corrugator supercilii. In one embodiment, at least a single unit dose may be administered to the corrugator supercilii, for example, two or more, three or more, four or more, or five or more unit doses may be administered. If desired, the corrugator supercilii proximal to one or both eyes may be treated.

[0115] When treating facial spasms, one or more (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, or all) additional muscles and / or regions thereof may be treated, wherein the one or more muscles and / or regions thereof are selected from the following: orbicularis oris (e.g., superior and / or inferior); zygomatic muscles (e.g., zygomatic major); nasal muscles; mentalis; platysma; frontalis; corrugator supercilii; buccinator; masseter; procerus; and lateral canthus. When treating facial spasms, one or more (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, or all) additional muscles and / or regions thereof may be treated, where the one or more muscles and / or regions thereof are selected from the following: orbicularis oris (e.g., orbicularis oris superior and / or orbicularis oris inferior); zygomaticus (e.g., zygomaticus major and / or zygomaticus minor); nasal muscles; mentalis; platysma; frontalis; corrugator supercilii; buccinator; masseter; procerus; and levator palpebrae superioris.

[0116] Preferably, one or more (eg, at least two, three, or four, or all) additional muscles and / or portions thereof selected from the following: corrugator supercilii, frontalis, zygomaticus major, buccinator, and masseter.

[0117] If the facial spasm is bilateral, the modified BoNT / A may be administered to any muscle and / or site on both sides of the patient's face. If the facial spasm is hemifacial spasm, the modified BoNT / A may be administered to any muscle and / or site on the affected side of the patient's face. At least a single unit dose may be administered to the muscle and / or site, for example, two or more (e.g., three or more, four or more, or five or more) unit doses may be administered.

[0118] The frontalis muscle may be the venter frontalis muscle.

[0119] The corrugator supercilii muscle may be a corrugator supercilii muscle.

[0120] In any aspect or embodiment of the invention relating to the treatment of blepharospasm, it is particularly preferred that the modified BoNT / A be administered by intramuscular injection at each of the following sites: a) Lateral superior orbicularis oculi muscle; b) medial orbicularis oculi superioris; and c) Lateral inferior orbicularis oculi muscle.

[0121] The (three) sites outlined in the immediately preceding paragraph may be referred to as the "minimal" sites for the treatment of blepharospasm.

[0122] In other words, in any aspect or embodiment of the invention relating to the treatment of blepharospasm, it is particularly preferred to administer the modified BoNT / A by intramuscular injection into each of the following muscles: a) the lateral orbicularis oculi superioris muscle affected by said blepharospasm; b) the medial orbicularis oculi superioris muscle affected by said blepharospasm; and c) The lateral inferior orbicularis oculi muscle affected by said blepharospasm.

[0123] In other words, in any aspect or embodiment of the invention relating to the treatment of blepharospasm, it is particularly preferred that the modified BoNT / A is administered by intramuscular injection into each of the following muscles: a) each lateral superior orbicularis oculi muscle affected by said blepharospasm; b) each medial superior orbicularis oculi muscle affected by said blepharospasm; and c) Each lateral inferior orbicularis oculi muscle affected by said blepharospasm.

[0124] For example, when treating blepharospasm, it is particularly preferred that the method comprises: a) administering up to 2 unit doses of modified BoNT / A into the lateral superior orbicularis oculi muscle proximal to the patient's first eye; b) administering a unit dose of the modified BoNT / A to the medial orbicularis oculi superior muscle proximal to the first eye of the patient; and c) administering a unit dose of the modified BoNT / A into the inferior lateral orbicularis oculi muscle proximal to the patient's first eye.

[0125] In other words, when treating blepharospasm, it is particularly preferred that the method comprises: a) administering up to 2 unit doses of modified BoNT / A to each lateral superior orbicularis oculi muscle affected by said blepharospasm; b) administering a unit dose of modified BoNT / A to each medial orbicularis oculi superior muscle affected by said blepharospasm; and c) administering a unit dose of modified BoNT / A to each inferior lateral orbicularis oculi muscle affected by said blepharospasm.

[0126] For example, when treating blepharospasm, it is particularly preferred that the method comprises: a) administering a two unit dose of modified BoNT / A into the lateral orbicularis oculi superior muscle proximal to the patient's first eye; b) administering a single unit dose of the modified BoNT / A to the medial orbicularis oculi superior muscle proximal to the first eye of the patient; and c) administering a single unit dose of the modified BoNT / A into the inferior lateral orbicularis oculi muscle proximal to the subject's first eye.

[0127] In other words, when treating blepharospasm, it is particularly preferred that the method comprises: a) administering two unit doses of modified BoNT / A to each lateral superior orbicularis oculi muscle affected by said blepharospasm; b) administering a single unit dose of modified BoNT / A to each medial orbicularis oculi superior muscle affected by said blepharospasm; and c) administering a single unit dose of modified BoNT / A to each inferior lateral orbicularis oculi muscle affected by said blepharospasm.

[0128] References to the "upper" orbicularis oculi muscle refer to the orbicularis oculi muscle of the upper eyelid. Similarly, references to the "lower" orbicularis oculi muscle refer to the orbicularis oculi muscle of the lower eyelid.

[0129] Those of skill in the art understand that the term "medial" (e.g., in an anatomical context) means toward the midline of the body. Similarly, those of skill in the art understand that the term "lateral" (e.g., in an anatomical context) means away from the midline of the body. Thus: - "Lateral" superior orbicularis oculi refers to the portion of the upper eyelid orbicularis oculi that is located away from the midline of the body (see, for example, positions (1) and (2) in Figure 8).

[0130] - The "medial" superior orbicularis oculi refers to the portion of the upper eyelid orbicularis oculi that is located closer to the midline of the body (see, for example, position (2) in Figure 8).

[0131] - The "lateral" inferior orbicularis oculi refers to the portion of the upper eyelid orbicularis oculi that is located away from the midline of the body (see, for example, position (4) in Figure 8).

[0132] The term "lateral superior orbicularis oculi muscle" may be used synonymously with the term "lateral portion of the orbicularis oculi muscle of the upper eyelid." The term "medial superior orbicularis oculi muscle" may be used synonymously with the term "lateral portion of the orbicularis oculi muscle of the upper eyelid." The term "lateral inferior orbicularis oculi muscle" may be used synonymously with the term "lateral portion of the orbicularis oculi muscle of the lower eyelid."

[0133] The orbicularis oculi muscle includes the "superior anterior tarsal plate" and the "anterior orbital septum" (both can be injected).

[0134] Thus, administration to the lateral orbicularis oculi superior muscle can mean administration to the lateral "pretarsal" or lateral "prepalpebral" orbicularis oculi superior muscle.

[0135] Administration to the medial superior orbicularis oculi muscle can mean administration to the medial superior "super pretarsal" or medial superior "prepalpebral" orbicularis oculi muscle.

[0136] Administration to the lateral inferior orbicularis oculi muscle can mean administration to the lateral inferior "pretarsal" or lateral inferior "prepalpebral" orbicularis oculi muscle.

[0137] When administered to the lateral superior orbicularis oculi muscle, the unit dose is preferably administered anterior to the septum (i.e., the lateral "pre-palpebral" superior orbicularis oculi muscle). When administered to the medial superior orbicularis oculi muscle, the unit dose is preferably administered anterior to the septum (i.e., the medial superior "pre-palpebral" orbicularis oculi muscle). When administered to the lateral inferior orbicularis oculi muscle, the unit dose is preferably administered anterior to the septum (i.e., the lateral inferior "pre-palpebral" orbicularis oculi muscle).

[0138] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose into the orbicularis oris superioris muscle; (ii) 1 unit dose into the inferior orbicularis oris muscle; (iii) 1 unit dose into the zygomatic major muscle; (iv) 1 unit dose into the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; (vi) 1 unit dose into the mentalis muscle; (vii) 1 unit dose into the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii; (ix) 1 unit dose into the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) into the masseter muscle; (xi) 1 unit dose into the procerus muscle; (xii) one unit dose into the bridge of the nose; and / or (xiii) One unit dose into the levator palpebrae superioris.

[0139] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) into the corrugator supercilii; (ii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; and / or (iii) One unit dose into the procerus muscle.

[0140] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) per corrugator supercilii muscle (e.g., affected by said blepharospasm); (ii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; and / or (iii) One unit dose into the procerus muscle.

[0141] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; or 1 unit dose to the procerus muscle; and / or (ii) Up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle.

[0142] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) per corrugator supercilii muscle (e.g., muscle affected by blepharospasm); or 1 unit dose per procercus muscle; and / or (ii) Up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle.

[0143] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle and / or 1 unit dose to the procerus muscle; and optionally, (ii) Up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle.

[0144] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit per corrugator supercilii muscle (e.g., muscle affected by blepharospasm) and / or 1 unit dose per procercus muscle; and optionally (ii) Up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle.

[0145] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to an additional muscle affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) into the corrugator supercilii; (ii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; and (iii) One unit dose into the procerus muscle.

[0146] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to said additional muscle affected by blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) per corrugator supercilii muscle (e.g., the muscle affected by said blepharospasm); and (ii) up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle; and (iii) One unit dose into the procerus muscle.

[0147] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to an additional muscle affected by said blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle or 1 unit dose to the procerus muscle; and (ii) Up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle.

[0148] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to said additional muscle affected by blepharospasm according to the following dosing schedule: (i) up to 2 unit doses (preferably 1 unit dose) per corrugator supercilii muscle (e.g., muscle affected by blepharospasm) or 1 unit dose per procercus muscle; and (ii) Up to 5 unit doses (preferably 1 unit dose) into the frontalis muscle.

[0149] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose into the corrugator supercilii; (ii) 2 unit doses into the frontalis muscle (e.g., 1 unit dose per affected eye); and / or (iii) One unit dose into the procerus muscle.

[0150] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose per corrugator supercilii (the muscle affected by facial spasm); (ii) 2 unit doses into the frontalis muscle (e.g., 1 unit dose per affected eye); and / or (iii) One unit dose into the procerus muscle.

[0151] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) one unit dose to the corrugator supercilii muscle or one unit dose to the procerus muscle; and / or (ii) 2 unit doses into the frontalis muscle (e.g., 1 unit dose per affected eye).

[0152] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said blepharospasm according to the following dosing schedule: (i) one unit dose per corrugator supercilii muscle (e.g., the muscle affected by blepharospasm) or one unit dose per procercus muscle; and / or (ii) 2 unit dose into the frontalis muscle.

[0153] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to an additional muscle affected by said blepharospasm according to the following dosing regimen: (i) 1 unit dose into the corrugator supercilii; (ii) 2 unit doses into the frontalis muscle (e.g., 1 unit dose per affected eye); and / or (iii) One unit dose into the procerus muscle.

[0154] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to said additional muscle affected by blepharospasm according to the following dosing regimen: (i) 1 unit dose per corrugator supercilii muscle (e.g., the muscle affected by blepharospasm); (ii) 2 unit doses into the frontalis muscle (e.g., 1 unit dose per affected eye); and (iii) One unit dose into the procerus muscle.

[0155] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to an additional muscle affected by said blepharospasm according to the following dosing regimen: (i) 1 unit dose to the corrugator supercilii muscle or 1 unit dose to the procerus muscle; and (ii) 2 unit doses into the frontalis muscle (e.g., 1 unit dose per affected eye).

[0156] In other words, in any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering a modified BoNT / A to said additional muscle affected by blepharospasm according to the following dosing regimen: (i) 1 unit dose per corrugator supercilii (e.g., the muscle affected by blepharospasm) or 1 unit dose per procercus muscle; and (ii) 2 unit dose into the frontalis muscle.

[0157] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the orbicularis oris superior muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris inferior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0158] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the inferior orbicularis oris muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0159] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the zygomatic major muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the superior orbicularis oris muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses to the frontalis muscle (preferably 1 unit dose); (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses to the corrugator supercilii muscle (preferably 1 unit dose); (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses to the masseter muscle (preferably 1 unit dose); (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0160] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the zygomatic minor muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the superior orbicularis oris muscle; (iv) up to 5 unit doses to the frontalis muscle (preferably 1 unit dose); (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses to the corrugator supercilii muscle (preferably 1 unit dose); (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses to the masseter muscle (preferably 1 unit dose); (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0161] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering up to 5 unit doses (preferably 1 unit dose; more preferably 2 unit doses; most preferably 3 unit doses) to the frontalis muscle; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris inferior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) 1 unit dose to the orbicularis oculi superior muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0162] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the mentalis muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the superior orbicularis oris muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0163] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the platysma muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the superior orbicularis oris muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0164] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) 1 unit dose to the superior orbicularis oris muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0165] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the buccinator muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the zygomatic major muscle; (iii) 1 unit dose to the zygomatic minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the superior orbicularis oris muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0166] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering 1 to up to 2 unit doses to the masseter muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris inferior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) 1 unit dose to the orbicularis oris superior muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0167] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the procerus muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris inferior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the orbicularis oris superior muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0168] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the nasal bridge; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris inferior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the orbicularis oris superior muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0169] In any aspect or embodiment of the invention directed to the treatment of blepharospasm, the invention may further comprise administering one unit dose to the levator palpebrae superioris muscle; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said blepharospasm according to the following dosing schedule: (i) 1 unit dose to the inferior orbicularis oris muscle; (ii) 1 unit dose to the zygomatic major muscle; (iii) 1 unit dose to the zygomatic minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal bridge; and / or (xii) 1 unit dose to the superior orbicularis oris muscle.

[0170] As outlined above, in aspects of the invention directed to the treatment of typical hemifacial spasm, the invention may comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said typical hemifacial spasm according to the following dosing schedules: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the orbicularis oris inferior muscle; (iii) 1 unit dose to the zygomaticus major muscle; (iv) 1 unit dose to the zygomaticus minor muscle; (v) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (vi) 1 unit dose to the mentalis muscle; (vii) 1 unit dose to the platysma muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (ix) 1 unit dose to the buccinator muscle; (x) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (xi) 1 unit dose to the procerus muscle; (xii) 1 unit dose to the nasal muscle, and / or (xiii) 1 unit dose to the levator palpebrae superioris muscle.

[0171] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the orbicularis oris superior muscle affected by said typical hemifacial spasm; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris inferior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0172] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the inferior orbicularis oris muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0173] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the zygomatic major muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the superior orbicularis oris muscle; (ii) 1 unit dose to the zygomaticus minor muscle; (iii) inferior orbicularis oris muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0174] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the zygomatic minor muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the orbicularis oculi inferior muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0175] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering up to 5 unit doses (preferably 1 unit dose) to said frontalis muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) 1 unit dose to the orbicularis oris inferior muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0176] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the mentalis muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the superior orbicularis oris muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the inferior orbicularis oris muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0177] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the platysma muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the orbicularis oris inferior muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0178] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) 1 unit dose to the orbicularis oris inferior muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0179] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the buccinator muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the orbicularis oris inferior muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0180] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering up to 2 unit doses (preferably 1 unit dose) to the masseter muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) 1 unit dose to the orbicularis oris inferior muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0181] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the procerus muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the orbicularis oris inferior muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0182] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further include: administering one unit dose to the nasal muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the orbicularis oris superior muscle; (ii) 1 unit dose to the zygomaticus major muscle; (iii) 1 unit dose to the zygomaticus minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the orbicularis oris inferior muscle; and / or (xii) 1 unit dose to the levator palpebrae superioris muscle.

[0183] In any aspect or embodiment of the invention directed to the treatment of typical hemifacial spasm, the invention may further comprise: administering one unit dose to the levator palpebrae superioris muscle affected by said typical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, or 12 or more) additional muscles affected by said typical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the superior orbicularis oris muscle; (ii) 1 unit dose to the zygomatic major muscle; (iii) 1 unit dose to the zygomatic minor muscle; (iv) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (v) 1 unit dose to the mentalis muscle; (vi) 1 unit dose to the platysma muscle; (vii) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (viii) 1 unit dose to the buccinator muscle; (ix) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (x) 1 unit dose to the procerus muscle; (xi) 1 unit dose to the nasal muscle; and / or (xii) 1 unit dose to the inferior orbicularis oris muscle.

[0184] In aspects of the invention directed to the treatment of atypical hemifacial spasm, as outlined above, the invention may comprise administering one or more unit doses of modified BoNT / A to one or more additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (i) 1 unit dose to the zygomatic major muscle; (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi inferior muscle; (xii) 1 unit dose to the medial palpebral muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0185] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the zygomatic major muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral inferior orbicularis oculi muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0186] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the zygomatic minor muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic major muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial palpebral superior muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0187] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further include: (i) administering up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) 1 unit dose to the zygomatic major muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral inferior orbicularis oculi muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0188] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the mentalis muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the zygomatic major muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral inferior orbicularis oculi muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0189] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the platysma muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the zygomatic minor muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral inferior orbicularis oculi muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0190] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering up to two unit doses (preferably one unit dose) to the corrugator supercilii muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, or thirteen or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) 1 unit dose to the zygomatic major muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0191] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the buccinator muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the zygomatic major muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral inferior orbicularis oculi muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0192] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering up to two unit doses (preferably one unit dose) to the masseter muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, or thirteen or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) 1 unit dose to the zygomatic major muscle; (ix) 1 unit dose to the proximal canalis muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial palpebrae superior muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0193] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the procerus muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the zygomatic major muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral inferior orbicularis oculi muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0194] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further include: (i) administering one unit dose to the nasal muscle affected by said atypical hemifacial spasm; and, optionally, administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the zygomatic major muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial palpebral superior muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0195] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the lateral orbicularis oculi superioris muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the zygomatic major muscle; (xii) 1 unit dose to the medial palpebrae superioris muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0196] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the medial orbicularis oculi superior muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the zygomatic major muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0197] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the lateral inferior orbicularis oculi muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superioris muscle; (xii) 1 unit dose to the medial palpebrae superioris muscle; (xiii) 1 unit dose to the zygomatic major muscle; and / or (xiv) 1 unit dose to the levator palpebrae superioris muscle.

[0198] In any aspect or embodiment of the invention directed to the treatment of atypical hemifacial spasm, the invention may further comprise: (i) administering one unit dose to the levator palpebrae superioris muscle affected by said atypical hemifacial spasm; and optionally administering one or more unit doses of modified BoNT / A to one or more (e.g., 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 11 or more, 12 or more, or 13 or more) additional muscles affected by said atypical hemifacial spasm according to the following dosing schedule: (ii) 1 unit dose to the zygomatic minor muscle; (iii) up to 5 unit doses (preferably 1 unit dose) to the frontalis muscle; (iv) 1 unit dose to the mentalis muscle; (v) 1 unit dose to the platysma muscle; (vi) up to 2 unit doses (preferably 1 unit dose) to the corrugator supercilii muscle; (vii) 1 unit dose to the buccinator muscle; (viii) up to 2 unit doses (preferably 1 unit dose) to the masseter muscle; (ix) 1 unit dose to the procerus muscle; (x) 1 unit dose to the nasal muscle; (xi) 1 unit dose to the lateral orbicularis oculi superior muscle; (xii) 1 unit dose to the medial superior palpebral muscle; (xiii) 1 unit dose to the lateral orbicularis oculi inferior muscle; and / or (xiv) 1 unit dose to the zygomatic major muscle.

[0199] The modified BoNT / A may be administered to the muscle and / or site of the present invention by any suitable means.

[0200] As an alternative to intramuscular injection, the modified BoNT / A may be administered subcutaneously, for example by subcutaneous injection, which may include medial and / or lateral injection into the junction between the anterior orbital septum and the orbital part of the superior and / or inferior orbicularis oculi muscles, if desired.

[0201] Most preferably, the modified BoNT / A is administered intramuscularly, for example, by intramuscular injection.

[0202] In accordance with the present invention, electromyographic control / electromyographic guidance may be used to assist in the administration of the modified BoNT / A.

[0203] The term "unit dose" may encompass one or more unit doses. For example, the term "unit dose" may mean up to 2 unit doses, up to 3 unit doses, up to 4 unit doses, or up to 5 unit doses. The term "unit dose" may also refer to a single unit dose.

[0204] A single unit dose may be administered to the affected muscle at one or more injection sites. When a single unit dose is administered to multiple injection sites, the unit dose may be divided (equally or unequally) between two or more injection sites. However, it is preferred that a single unit dose is administered per injection site.

[0205] The term "administering a single unit dose" means that substantially all of the single unit dose is administered. For example, a remainder of the unit dose (e.g., up to 1%, 0.1%, or 0.01%) may remain in the vial in which the modified BoNT / A is reconstituted. However, preferably, all of the single unit dose is administered (e.g., for one or more injection sites, preferably for each injection site). This definition applies equally to administration of two unit doses, three unit doses, etc.

[0206] The potency of the modified BoNT / A for use according to the present invention was measured using mouse LD 50 In said assay, one unit is determined to be the calculated median lethal dose (LD ) in mice. 50 ) in mice. 50 ).

[0207] A modified BoNT / A for use in the present invention comprises a BoNT / A light chain and translocation domain and a BoNT / B receptor binding domain (H C In the case of a modified BoNT / A comprising the above-mentioned nucleotide sequence (domain), the amount of modified BoNT / A corresponding to 1 unit in the above-mentioned assay is preferably 24.04 pg.

[0208] The term "up to," when used in connection with a value (e.g., up to 82,500 pg), means up to and including the recited value. Thus, for example, reference to administering "up to 82,500 pg" of modified BoNT / A includes not only administration of 82,500 pg of modified BoNT / A, but also administration of less than 82,500 pg of modified BoNT / A.

[0209] The unit dose may be expressed in terms of an amount of modified BoNT / A, units of modified BoNT / A, or a combination thereof.

[0210] The total number of unit doses administered may be up to 20, 15, 10, 5, or 3. The total number of unit doses administered may be at least 3, 5, 10, or 15. The total number of unit doses administered may be 3 to 20, 4 to 16, or 5 to 12. In one embodiment, 5 unit doses are administered. In one embodiment, 6 unit doses are administered. In one embodiment, 10 unit doses are administered. In one embodiment, 12 unit doses are administered. In one embodiment, 15 doses are administered. Administration of a total of 12 unit doses is preferred, particularly for the treatment of blepharospasm, more particularly bilateral blepharospasm.

[0211] The modified BoNT / A may be administered by intramuscular injection to a total of 6, 7, 8, 9, 10, 11, or 12 sites in the patient's first eye. The modified BoNT / A may be administered by intramuscular injection to a total of 6, 7, 8, 9, 10, or 11 sites in the patient's first eye. Additionally or alternatively, the modified BoNT / A may be administered by intramuscular injection to a total of 6, 7, 8, 9, 10, 11, or 12 sites in the patient's second eye. The modified BoNT / A may be administered by intramuscular injection to a total of 6, 7, 8, 9, 10, or 11 sites in the patient's second eye.

[0212] In a preferred embodiment, the modified BoNT / A is administered by intramuscular injection to a total of six sites in the patient's first eye. Additionally or alternatively, the modified BoNT / A may be administered by intramuscular injection to a total of six sites, preferably in the patient's second eye.

[0213] When treating blepharospasm (e.g., preferably bilateral blepharospasm), it is preferred to administer up to 12 unit doses over the following sites: - 2 unit doses into the lateral superior orbicularis oculi muscle of the affected eye (e.g., 2 unit doses into said area in each eye for a total of 4 unit doses); - 1 unit dose into the medial superior orbicularis oculi muscle of the affected eye (e.g., 1 unit dose into said area in each eye for a total of 2 unit doses); - 1 unit dose into the lateral inferior orbicularis oculi muscle of the affected eye (e.g., 1 unit dose into said area in each eye for a total of 2 unit doses); - 1 unit dose into the procerus muscle; - 1 unit administered to the corrugator supercilii muscle proximal to the affected eye (e.g., 1 unit dose to that site in each eye for a total of 2 unit doses); and - 1 unit dose into the frontalis muscle proximal to the affected eye (e.g., 1 unit dose into said area in each eye for a total of 2 unit doses).

[0214] For example, the treatment may comprise a total of 12 unit dose injections over the abovementioned sites. It is preferred to administer 12 unit dose injections over the abovementioned sites (particularly for blepharospasm, more particularly the bilateral type).

[0215] When treating unilateral blepharospasm, it is advisable to administer up to 6 unit doses over the following areas: - 2 unit doses into the lateral superior orbicularis oculi muscle of the affected eye; - 1 unit dose into the medial superior orbicularis oculi muscle of the affected eye; - 1 unit dose into the lateral inferior orbicularis oculi muscle of the affected eye; - 1 unit dose into the procerus muscle; - 1 unit dose into the corrugator supercilii muscle proximal to the affected eye; and - 1 unit dose into the frontalis muscle proximal to the affected eye.

[0216] For example, the treatment may involve injection of a total of 6 unit doses over the above mentioned sites.

[0217] Optionally, the unit dose may be administered to only one (but not both) selected from the procerus and corrugator supercilii muscles.

[0218] The following injection sites are preferably included in the above-described treatment regimen (e.g., may be referred to as "minimal" injection sites): - 2 unit doses into the lateral superior orbicularis oculi muscle of the affected eye (e.g., 2 unit doses into the aforementioned area in each eye for a total of 4 unit doses); - 1 unit dose into the medial orbicularis oculi superior muscle of the affected eye (e.g., 1 unit dose into said area in each eye for a total of 2 unit doses); and - 1 unit dose into the outer inferior orbicularis oculi muscle of the affected eye (e.g., 1 unit dose into said area in each eye for a total of 2 unit doses).

[0219] The "smallest" injection site may be sufficient to administer the treatment, so that a total of 4 unit doses are administered per eye (e.g., for unilateral blepharospasm, a total of 4 unit doses). That said, the treatment may be extended to the procerus and / or corrugator supercilii (preferably the procerus or corrugator supercilii), with an additional unit dose for the procerus and / or an additional unit dose for the corrugator supercilii being added to the treatment plan. Additionally or alternatively, the treatment may be extended to the frontalis of the affected eye, with an additional unit dose (for the frontalis) being added per affected eye.

[0220] The skilled artisan may take into consideration cases where the patient has recently undergone (or will subsequently undergo) additional treatment with a Clostridial neurotoxin (e.g., unmodified BoNT), for example as part of a cosmetic surgery or treatment for another condition. Using routine techniques in the art, the skilled artisan may adapt the treatment regimen appropriately. Preferably, the present invention excludes treatment with an additional Clostridial neurotoxin (e.g., BoNT).

[0221] The modified BoNT / A of the present invention preferably has a longer duration of action when compared to unmodified BoNT / A (e.g., Dysport®), e.g., at least 5%, 10%, 25%, or 50% improvement in one or more symptoms of blepharospasm or hemifacial spasm compared to the one or more pre-treatment symptoms. The duration of action can be at least 1.25-fold, 1.5-fold, 1.75-fold, 2.0-fold, or 2.25-fold. The duration of action of the modified BoNT / A can be between 6 months and 9 months. For example, the duration of action can be at least: 4.5 months, 5.0 months, 5.5 months, 6 months, 6.5 months, 7.0 months, 7.5 months, 8 months, 8.5 months, or 9.0 months (from onset). In certain embodiments, the duration of action can be longer than 9.0 months.

[0222] Treatment may be repeated at an appropriate time period after administration of modified BoNT / A. Given that the duration of action is approximately twice that of unmodified BoNT / A (e.g., Dysport®), there is an appropriately longer period between subsequent administrations than when the patient is treated with unmodified BoNT / A (e.g., Dysport®). In the present invention, the patient may be re-administered BoNT / A at least 18 weeks, 20 weeks, 25 weeks, or 30 weeks after the previous administration. For example, in the present invention, the patient may be re-administered modified BoNT / A at least 18 weeks to 45 weeks, preferably 20 weeks to 35 weeks, after the previous administration.

[0223] As used herein, a "patient" may be a mammal, such as a human or other mammal. Preferably, "patient" refers to a human patient. A "patient" is preferably an adult patient, i.e., a patient who is at least 18 years of age. The terms "patient" and "patient" are used synonymously herein. Preferably, the patient has been diagnosed with facial dystonia (blepharospasm, typical hemifacial spasm, or atypical hemifacial spasm) according to the present invention.

[0224] A patient for treatment according to the invention may be a patient who is unsuitable for treatment with unmodified BoNT / A (e.g., of SEQ ID NO: 2). The patient may be a patient who is resistant to treatment with unmodified BoNT / A. Resistance may occur due to the development of an immune response against a clostridial neurotoxin, including the production by the patient of anti-clostridial neurotoxin antibodies.

[0225] As used herein, the term "treatment" or "treating" encompasses not only corrective treatment (treatment of a patient already suffering from a disease) but also prophylactic treatment (e.g., to prevent the onset of a disease). Preferably, as used herein, "treatment" or "treating" refers to corrective treatment. As used herein, the term "treatment" or "treating" refers to the disease and / or symptoms thereof.

[0226] An example of a clostridial neurotoxin produced by bacteria of the genus Clostridium is BoNT / A. Other examples of such clostridial neurotoxins include those produced by Clostridium baratii and C. butyricum, as well as those produced by Clostridium tetani (TeNT) and Clostridium botulinum (BoNT) serotypes B to G. The neurotoxins are highly potent and specific, and can poison nerves and other cells to which they are delivered. The clostridial toxins are some of the most potent toxins known. For example, botulinum neurotoxins have a median lethal dose (LD ) in mice ranging from 0.5 to 5 ng / kg, depending on the serotype. 50 ) value. Both tetanus toxin and botulinum toxin act by inhibiting the function of affected neurons, specifically the release of neurotransmitters. Botulinum toxin acts at the neuromuscular junction and inhibits cholinergic transmission in the peripheral nervous system, while tetanus toxin acts in the central nervous system.

[0227] In nature, clostridial neurotoxins (including BoNT / A) are synthesized as single-chain polypeptides by a proteolytic cleavage event that results in the formation of two polypeptide chains linked by a disulfide bond. Cleavage occurs at a specific cleavage site, often referred to as the active site, located between the cysteine ​​residues that provide the interchain disulfide bond. It is this two-chain form that is the active form of the toxin. The two chains are referred to as the heavy chain (H chain), with a molecular weight of approximately 100 kDa, and the light chain (L chain), with a molecular weight of approximately 50 kDa. The H chain is composed of an N-terminal cartwheel component (H chain), which is connected to the ...). N domain) and the C-terminal targeting component (H C The cleavage site is located between the L chain and the translocation domain component. C The H domain binds to a target neuron and the bound toxin is internalized into the cell by endosomes. N The domain allows the L chain to permeabilize the endosomal membrane and translocate into the cytosol, where it provides a protease function (also known as a non-cytotoxic protease).

[0228] Non-cytotoxic proteases act by proteolytic cleavage of intracellular transport proteins known as SNARE proteins (e.g., SNAP-25, VAMP or syntaxin) - see Gerald K (2002) “Cell and Molecular Biology“ (4th edition) John Wiley & Sons, Inc. The acronym SNARE is derived from Solute NSF Attachment Receptor, where NSF stands for N-ethylmaleimide sensitive factor. SNARE proteins are essential for intracellular vesicle fusion and thus for the secretion of molecules from cells via vesicular trafficking. The protease function is a zinc-dependent endopeptidase activity and shows high substrate specificity for SNARE proteins. Thus, once delivered to the desired target cells, non-cytotoxic proteases are able to inhibit cellular secretion from the target cells. The L-chain proteases of clostridial toxins are non-cytotoxic proteases that cleave SNARE proteins.

[0229] Given the ubiquitous nature of SNARE proteins, clostridial neurotoxins, such as botulinum toxin, have been used successfully in a wide range of therapeutic approaches.

[0230] For further details regarding the genetic basis of toxin production in Clostridium botulinum and Clostridium tetani, see Henderson et al (1997) in The Clostridia: Molecular Biology and Pathogenesis, Academic press.

[0231] As mentioned above, clostridial neurotoxins are composed of two polypeptide chains, a heavy chain (H chain) with a molecular weight of about 100 kDa and a light chain (L chain) with a molecular weight of about 50 kDa. The H chain contains a C-terminal targeting component (receptor binding domain or H C domain) and the N-terminal cartwheel component (H N domain), and

[0232] Clostridial neurotoxin domains are described in more detail below.

[0233] Examples of light chain reference sequences include: Botulinum neurotoxin type A: amino acid residues 1-448 Botulinum neurotoxin type B: amino acid residues 1-440

[0234] The above specific reference sequences should be considered as a guideline, as slight variations may occur depending on the subserotype. For example, US2007 / 0166332 (hereby incorporated by reference in its entirety) cites the following slightly different Clostridium sequences: Botulinum neurotoxin type A: amino acid residues M1 to K448 Botulinum neurotoxin type B: amino acid residues M1 to K441

[0235] A translocation domain is a fragment of the heavy chain of a clostridial neurotoxin approximately equal to the amino-terminal half of the heavy chain, or the domain corresponding to that fragment in an intact heavy chain.

[0236] Examples of translocation domain references include: Botulinum neurotoxin type A - amino acid residues (449-871) Botulinum neurotoxin type B - amino acid residues (441-858)

[0237] The above specific reference sequences should be considered as a guideline, as slight variations may occur depending on the subserotype. For example, US2007 / 0166332 (hereby incorporated by reference) cites the following slightly different Clostridium sequences: Botulinum neurotoxin type A - amino acid residues (A449~K871) Botulinum neurotoxin type B - amino acid residues (A442-S858)

[0238] In the context of the present invention, various BoNT / AHs containing a translocation domain N The regions may be useful in embodiments of the present invention.N The region is approximately 410 to 430 amino acids in length and contains the translocation domain. N Studies have shown that the full length of the region is not necessary for the translocation activity of the translocation domain. Thus, aspects of this embodiment include BoNT / AH polypeptides that include a translocation domain having a length of, for example, at least 350 amino acids, at least 375 amino acids, at least 400 amino acids, or at least 425 amino acids. N Other aspects of this embodiment may include a BoNT / AH domain that includes a translocation domain having a length of, for example, at most 350 amino acids, at most 375 amino acids, at most 400 amino acids, or at most 425 amino acids. N It may contain areas.

[0239] H N The term refers to naturally occurring BoNT / AH. N and modified BoNT / AH having non-naturally occurring amino acid sequences and / or artificial amino acid residues. N Preferably, the modified BoNT / AH N The moiety still exhibits the translocation function described above.

[0240] Clostridial neurotoxin receptor binding domain (H C ) Examples of reference sequences include: BoNT / A‐N872-L1296 BoNT / B‐E859-E1291

[0241] The Hc domain of clostridial neurotoxins (e.g., BoNT / A) is approximately 50 kDa. CC Domain and H CN It contains two distinct structural features called domains, each typically about 25 kDa. The amino acid residues involved in receptor binding are primarily H CC The H domain of natural Clostridial neurotoxins is thought to be located in CThe domain may comprise about 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. Chem. 268: 15, pp. 11188-11192; Rummel A (2007) PNAS 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.

[0242] H CN Example domains (references) include: Botulinum neurotoxin type A - amino acid residues (872-1110) Botulinum neurotoxin type B - amino acid residues (859-1097)

[0243] The above sequence positions may vary slightly depending on the serotype / subserotype, and further H CN Example domains (references) include: Botulinum neurotoxin type A - amino acid residues (874-1110) Botulinum neurotoxin type B - amino acid residues (861-1097)

[0244] H CC Examples of domains include: Botulinum neurotoxin type A - amino acid residues (Y1111 to L1296) Botulinum neurotoxin type B - amino acid residues (Y1098-E1291)

[0245] L chain and H N The domains (optionally including a complete or partial activation loop, e.g., if the modified BoNT / A is in a single-chain form, including a complete activation loop, and if it is in a two-chain form, including a truncated / partial activation loop) are collectively referred to as the LH N The LH may be referred to as a domain. N The domain is therefore further C It does not include the domain.

[0246] WO2017 / 191315A1 (hereby incorporated by reference) teaches modified BoNT / A and methods for their preparation and manufacture. Thus, the botulinum neurotoxin A (BoNT / A) light chain and translocation domain (BoNT / AH) for use in the present invention are N ) and the BoNT / B receptor binding domain (H C The modified BoNT / A, including the AFG2 domain, may be one taught in WO 2017 / 191315A1.

[0247] As used herein, the term "modified BoNT / A" or "chimeric clostridial neurotoxin" or "chimeric neurotoxin" refers to a chimeric clostridial neurotoxin that is a ... N domain) and a receptor-binding domain (H) derived from a second different Clostridial neurotoxin serum. C Specifically, the modified BoNT / A used 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 modified BoNT / A comprises the BoNT / ALH domain. N The domain is BoNT / BH CThe modified BoNT / A of the present invention may be referred to as a chimeric botulinum neurotoxin. The modified BoNT / A is also referred to herein as a "BoNT / AB", "mrBoNT / AB" or "BoNT / AB chimera".

[0248] L chain and H N The domains (optionally including a complete or partial activation loop, e.g., if the modified BoNT / A is in single-chain form, including a complete activation loop, and if it is in two-chain form, including a truncated / partial activation loop) are collectively referred to as the LH N The LH may be referred to as a domain. N The domain is therefore further C It does not include the domain.

[0249] The modified BoNT / A is essentially a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N domain) and the BoNT / B receptor binding domain (H C domain).

[0250] As used in this context, the term "consists essentially of" means that the modified BoNT / A does not further comprise one or more amino acid residues that confer additional functionality to the polypeptide, e.g., when administered to a patient. In other words, the modified BoNT / A does not further comprise one or more amino acid residues that confer additional functionality to the polypeptide, e.g., when administered to a patient. N domain) and the BoNT / B receptor binding domain (H C A polypeptide "consisting essentially of" is a polypeptide that ... N domain) and the BoNT / B receptor binding domain (H C The polypeptide may further comprise one or more additional amino acid residues (residues of the corresponding domain), provided that the one or more additional amino acid residues do not confer additional functionality to the polypeptide, e.g., when administered to a patient. Additional functionality may include enzymatic activity, binding activity and / or any physiological activity.

[0251] The modified BoNT / A may include a non-clostridial neurotoxin sequence in addition to any clostridial neurotoxin sequence, so long as the non-clostridial neurotoxin sequence does not destroy the ability of the modified BoNT / A to achieve its own therapeutic effect. Preferably, the non-clostridial neurotoxin sequence does not have catalytic activity, such as enzymatic activity. In one embodiment, the modified BoNT / A of the present invention does not include a non-clostridial catalytic activity domain. In one embodiment, the modified BoNT / A does not include an additional catalytic activity 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 protein cellular receptor, such as an integral membrane protein. Examples of cellular receptors can be found 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 His tag. In one embodiment, a modified BoNT / A of the invention does not contain a label or a site for label attachment, such as a sortase acceptor or donor site.

[0252] Preferably, the modified BoNT / A 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).

[0253] The modified BoNT / A comprises a light chain capable of exhibiting non-cytotoxic proteases and capable of cleaving SNARE proteins in the cytoplasm of target neurons. Cell-based and in vivo assays may be used to determine whether a Clostridial neurotoxin comprising an L chain and a functional cell binding domain and translocation domain has non-cytotoxic protease activity. Assays such as the digit abduction score (DAS) assay, the dorsal root ganglion (DRG) assay, the spinal cord neuron (SCN) assay, and the mouse phrenic nerve hemidiaphragm (PNHD) assay are routine in the art. Suitable assays for determining non-cytotoxic protease activity may be those described in Aoki KR, Toxicon 39: 1815-1820; 2001or Donaldet al (2018), Pharmacol Res Perspect, e00446, 1-14, which are incorporated herein by reference.

[0254] When administered to a patient, the modified BoNT / A is preferably in an active dichain form, with the light and heavy chains linked by a disulfide bond. When BoNT / A (e.g., modified BoNT / A) is defined herein as a polypeptide sequence (SEQ ID NO:1), the L chain portion of said sequence (SEQ ID NO:1) may constitute the first chain of a dichain Clostridial neurotoxin (e.g., a dichain modified BoNT / A), and the H chain portion of said sequence (SEQ ID NO:1) may constitute the first chain of a dichain Clostridial neurotoxin (e.g., a dichain modified BoNT / A). N Domain and H CThe domains together may constitute the second chain of a dichain clostridial neurotoxin (e.g., dichain modified BoNT / A), where the first and second chains are linked together by disulfide bonds. Those skilled in the art will understand that the protease may cleave at one or more positions in the activation loop of the clostridial neurotoxin (e.g., modified BoNT / A), preferably at two positions in the activation loop. When cleavage occurs at one or more positions (preferably at two positions) in the activation loop, a small fragment of the C-terminal L chain portion of the sequence may be absent from the dichain clostridial neurotoxin sequence (e.g., dichain modified BoNT / A). With this in mind, the sequence of the dichain clostridial neurotoxin (e.g., dichain modified BoNT / A) may differ slightly from the sequence of the corresponding single-chain clostridial neurotoxin (e.g., single-chain modified BoNT / A). The small fragment may be 1 to 15 amino acids. In particular, in one embodiment, when Lys-C is used to convert a single-chain modified BoNT / A to a two-chain modified BoNT / A, the subfragment of the C-terminal L-chain portion of the absent sequence may be SEQ ID NO: 9 or 10.

[0255] Most preferably, the modified BoNT / A used in the present invention comprises a BoNT / A light chain and a translocation domain (BoNT / ALH N domain) and BoNT / BH C The BoNT / ALH domain may include N The domain is BoNT / BH C The modified BoNT / A is also referred to herein as a "BoNT / AB" or a "BoNT / AB chimera."

[0256] LH N The C-terminal amino acid residues of the domain are the same as those of the LH domain of BoNT / A. N and H C Separate domains 3 10 may correspond to the first amino acid residue of the helix, and C The N-terminal amino acid residues of the domain are the LH N Domain and H C Separate domains 3 10It may correspond to the second amino acid residue of the helix.

[0257] An example of a BoNT / A polypeptide sequence is provided as SEQ ID NO:2 (including the two-chain version of SEQ ID NO:2).

[0258] An example of a BoNT / B polypeptide sequence is provided as SEQ ID NO: 8 (UniProt Accession No. B1INP5).

[0259] As used herein, the term "BoNT / A LH N and H C Separate domains 3 10 The reference to "the first amino acid residue of the helix" refers to the LH N and H C Separate domains 3 10 It refers to the N-terminal residue of the helix.

[0260] The “LH of BoNT / B” N and H C Separate domains 3 10 The reference to "the second amino acid residue of the helix" is N and H C Separate domains 3 10 It refers to the amino acid residue following the N-terminal residue of the helix.

[0261] "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 of the helix are arranged in a right-handed helix with each full turn completed by three residues and ten atoms separating the intramolecular hydrogen bonds. Each amino acid corresponds to a 120° turn of the helix (i.e. the helix has three residues per turn), has a translation of 2.0 Å (=0.2 nm) along the helix axis, and has ten atoms in the ring formed by making hydrogen bonds. 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. This i+3→i hydrogen bond sequence is repeated three times.10 Define a helix. 3 10 A helix is ​​a standard concept in structural biology with which those of skill in the art are familiar.

[0262] These three 10 A helix corresponds to the four residues that form the actual helix plus two cap (or transition) residues, one at each end of these four residues. N Domain and H C Separate domains 3 10 The term "helix" consists of those six residues.

[0263] Through structural analysis and sequence alignment, LH N Domain and H C Separate domains 3 10 The three helices were identified. 10 The helix is ​​located at its N-terminus (i.e., the LH N The C-terminal part of the domain is surrounded by α-helices, and the C-terminus (i.e., the H C The N-terminal part of the domain is surrounded by β-strands. 10 The first (N-terminal) residue of the helix (the cap or transition residue) also corresponds to the C-terminal residue of this α-helix.

[0264] LH N Domain and H C Separate domains 3 10 The helices can be determined, for example, from published 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.

[0265] For example, 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 / ) were used to identify each LH in other neurotoxins. N Domain and each H N Separating the domain and 10 The positions of the helices may also be determined. In particular, the region around the "HN / HCN" junction may be highly structurally conserved, making it an ideal region for superimposing different serotypes.

[0266] For example, the following methodology was used to identify other neurotoxins: 10 The sequence of the helix can be determined. 1. The structural homology modeling tool LOOP (http: / / loopp.org) can be used to obtain predicted structures of other BoNT serotypes based on the BoNT / A1 crystal structure (3BTA.pdb); 2. The structure (pdb) file thus obtained may be edited to contain only the N-terminus of the HCN domain and the preceding approximately 80 residues (part of the HN domain), thus preserving the structurally highly conserved "HN / HCN" region; 3. Each serotype can be superimposed onto the 3BTA.pdb structure using the protein superposition server SuperPose (http: / / wishart.biology.ualberta.ca / superpose / ); 4. Inspect the overlapping pdb files and identify the 3′ end of the start of the HC domain of BoNT / A1. 10 The helices can be identified and the corresponding residues in other serotypes identified. 5. Other BoNT serotype sequences may be aligned with Clustal Omega to ensure that corresponding residues are correct.

[0267] LH determined by this method N , H C and 3 10 Examples of helical domains are shown below. [Table A]

[0268] Using structural analysis and sequence alignment, LH N Domain and H C Separate domains 3 10 The helix followed by a β-strand is a conserved structure in all botulinum and tetanus neurotoxins and is the LH N Domain and H C Separate domains 3 10 It was found to begin at the eighth residue when starting from the first residue of the helix (eg, BoNT / A1 residue 879).

[0269] BoNT / AB chimera is a BoNT / B derived H C LH from BoNT / A covalently bound to the domain N domain, where the LH N The C-terminal amino acid residue of the domain is the H C The H corresponds to the 8th amino acid residue from the N-terminus of the β chain located at the beginning (N-terminus) of the domain. CThe N-terminal amino acid residue of the domain is the H C It corresponds to the seventh amino acid residue from the N-terminus of the β chain located at the beginning (N-terminus) of the domain.

[0270] BoNT / AB chimera is a BoNT / B derived H C LH from BoNT / A covalently bound to the domain N domain, where the LH N The C-terminal amino acid residue of the LH domain of BoNT / A N corresponding to the C-terminal amino acid residue of the α-helix located at the end (C-terminus) of the domain, C The N-terminal amino acid residue of the LH domain of BoNT / B N It corresponds to an amino acid residue that is adjacent at the C-terminus to the C-terminal amino acid residue of the α-helix located at the end (C-terminus) of the domain.

[0271] The rationale for the design process of the BoNT / AB chimera is to ensure that the secondary structure is intact, thereby attempting to minimize any changes to the tertiary structure and function of each domain. 10 It is hypothesized that not disrupting the four central amino acid residues of the helix ensures an optimal conformation for the chimeric neurotoxin, thereby allowing the chimeric neurotoxin to maximize its efficacy. Indeed, surprisingly, the 3 10 The first amino acid residue of the helix and the 3 10 Retaining only the second amino acid residue after the helix not only allows for the production of a soluble and functional BoNT / AB, but also leads to superior properties over other BoNT / AB chimeras, particularly increased potency, increased safety factor and / or longer duration of action (as well as increased safety factor and / or duration of action compared to unmodified BoNT / A).

[0272] The BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BHc domain may be a modified BoNT / A light chain, a modified BoNT / A translocation domain, and / or a modified BoNT / BHc domain, or a derivative thereof, including, but not limited to, those described below. C The domain or derivative may be selected from the group consisting of the BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain. C The BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain may contain one or more amino acids that are modified as compared to the naturally occurring (unmodified) form of the domain. C The modified BoNT / A light chain, BoNT / A translocation domain, and / or BoNT / BHc domain may contain one or more inserted amino acids that are not present in the naturally occurring (unmodified) form of the domain. By way of example, a modified BoNT / A light chain, a modified BoNT / A translocation domain, and / or a modified BoNT / BHc domain may be a natural (unmodified) BoNT / A light chain, a BoNT / A translocation domain, and / or a BoNT / BHc domain. C The BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain may have a modified amino acid sequence in one or more of the domains compared to the domain sequence. Such modifications may alter its functional aspects, such as its biological activity or persistence. Thus, in one embodiment, the BoNT / A light chain, the BoNT / A translocation domain, and / or the BoNT / BH domain may have a modified amino acid sequence in one or more of the domains compared to the domain sequence. C The domains may be modified BoNT / A light chains, BoNT / A translocation domains, and / or BoNT / BH domains. C domain, or modified BoNT / A light chain, BoNT / A translocation domain, and / or BoNT / BH C It is a derivative of the domain.

[0273] Modifications that alter binding to target neurons, such as native (unmodified) BoNT / BH C The modified BoNT / BH domain exhibits higher or lower binding affinity compared to the C The BoNT / BH may have one or more domains. CSuch modifications in the H domain include those that alter the binding to ganglioside receptors and / or protein receptors of the target neuronal cells. C Modifications of residues in the ganglioside binding site or protein (e.g., synaptotagmin) binding site of the domain may be included. Examples of such modified neurotoxins are described in WO2006 / 027207 and WO2006 / 114308, both of which are incorporated herein by reference in their entirety.

[0274] A modified light chain may have one or more modifications in its amino acid sequence, such as modifications in the substrate binding domain or catalytic domain that may alter or modify the SNARE protein specificity of said modified light chain, preferably with the proviso that said modifications do not catalytically inactivate said light chain. Examples of such modified neurotoxins are described in WO2010 / 120766 and US2011 / 0318385, both of which are incorporated herein by reference in their entirety.

[0275] BoNT / A-derived LH N The domain may correspond to amino acid residues 1 to 872 of SEQ ID NO:2, 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:2, or a polypeptide sequence having at least 80%, 90%, or 95% sequence identity thereto. N The domain corresponds to amino acid residues 1 to 872 of SEQ ID NO:2.

[0276] BoNT / B-derived H C The domain may correspond to amino acid residues 860 to 1291 of SEQ ID NO:8, or a polypeptide sequence having at least 70% sequence identity thereto. CThe domain may correspond to amino acid residues 860 to 1291 of SEQ ID NO:8, 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:8.

[0277] Preferably, the BoNT / AB chimera is BoNT / A1LH N Domains and BoNT / B1H C More preferably, the LH N The H domain corresponds to amino acid residues 1 to 872 of BoNT / A1 (SEQ ID NO: 2). C The domain corresponds to amino acid residues 860 to 1291 of BoNT / B1 (SEQ ID NO:8).

[0278] Most preferably, BoNT / BH C The H domain further has the effect of increasing the binding affinity of the BoNT / B neurotoxin to human Syt2 as compared to the native BoNT / B sequence. CC The subdomain comprises at least one amino acid residue substitution, insertion, indel, or deletion. Suitable amino acid residue substitutions, insertions, indels, or deletions in the BoNT / BHcc subdomain are disclosed in WO2013 / 180799 and WO2016 / 154534, both of which are incorporated herein by reference.

[0279] Substitutions, insertions, indels or deletions of suitable amino acid residues in the BoNT / BHcc subdomain may include 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.

[0280] The substitution, insertion, indel or deletion of suitable amino acid residues in the BoNT / BHcc subdomain may include 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 S 1199Y, E1191Q 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.

[0281] The substitutions, insertions, indels or deletions of suitable amino acid residues in the BoNT / BHcc subdomain can also include a combination of the three substitution mutations E1191M, S1199W, and W1178Q.

[0282] Preferably, the substitution, insertion, indel or deletion of amino acid residues in the BoNT / B Hcc subdomain comprises a combination of two substitution mutations, E1191M and S1199Y. Such modifications are present, for example, in modified BoNT / A (e.g., BoNT / AB chimera) of SEQ ID NO:5 and SEQ ID NO:6. E1191M may correspond to position 1204 and S1199Y may correspond to position 1212 of SEQ ID NO:6. Thus, SEQ ID NO:6 may comprise 1204M and 1212Y.

[0283] The modification may be a modification relative to the unmodified BoNT / B shown as SEQ ID NO:8, where the numbering of the amino acid residues is determined by alignment with SEQ ID NO:8. Since the presence of a methionine residue at position 1 of SEQ ID NO:8 is optional (as are the SEQ ID NOs corresponding to the modified BoNT / A polypeptides described herein), the skilled artisan will take into account the presence / absence of the methionine residue when determining the numbering of the amino acid residues. For example, if SEQ ID NO:8 contains a methionine, the numbering of the position will be as defined above (e.g., E1191 becomes E1191 in SEQ ID NO:8). Alternatively, if a methionine is not present in SEQ ID NO:8, the numbering of the amino acid residue should be modified by -1 (e.g., E1191 becomes E1190 in SEQ ID NO:8). Thus, the methionine amino acid residue at position 1 of the polypeptide sequence of the modified BoNT / A may be optional or absent. Similar considerations apply to the presence / absence of a methionine at position 1 in other polypeptide sequences described herein, and one of skill in the art will readily determine the correct numbering of the amino acid residues using routine techniques in the art.

[0284] A modified BoNT / A 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: 3 to 7. For example, a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to a polypeptide sequence selected from SEQ ID NOs: 3 to 7. Preferably, a modified BoNT / A used in the present invention may comprise a polypeptide sequence selected from SEQ ID NOs: 3 to 7 (and more preferably, consists of said polypeptide sequence).

[0285] It is preferred that the modified BoNT / A comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 6. For example, a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 6. Most preferably, the modified BoNT / A used in the present invention may comprise (and more preferably consist of) SEQ ID NO: 6.

[0286] As used herein, the term "deletion" refers to the removal of one or more amino acid residues in a polypeptide without replacing the amino acid residue or 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 has x-1 amino acid residues.

[0287] As used herein, the term "indel" refers to the deletion of one or more amino acid residues in a polypeptide and the insertion of a different number of amino acid residues (either more or fewer amino acid residues) at the site of deletion compared to the number of amino acid residues deleted. Thus, for example, in the case of an indel in which 2 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.

[0288] As used herein, the term "substitution" 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 a polypeptide sequence having (for example) x amino acid residues, the resulting polypeptide also has x amino acid residues. Preferably, the substitution is at a single amino acid position.

[0289] As used herein, the term "insertion" refers to the addition of one or more amino acid residues to a polypeptide without deleting one or more amino acid residues of 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.

[0290] Methods for modifying proteins by substitution, insertion or deletion of amino acid residues are known in the art. By way of example, amino acid modifications can be introduced by modification of a DNA sequence encoding BoNT / A (e.g., encoding unmodified BoNT / A). This can be achieved using standard molecular cloning techniques, for example, by site-directed mutagenesis, where a short stretch of DNA (oligonucleotide) encoding the desired amino acid replaces the original coding sequence using a polymerase enzyme, or by inserting / deleting a part of the gene using various enzymes (e.g., ligases and restriction endonucleases). Alternatively, the modified gene sequence can be chemically synthesized. Typically, the modification can be performed by either modifying a nucleic acid encoding a naturally occurring Clostridial neurotoxin (or a part thereof) such that the modified BoNT / A (or a part thereof) encoded by said nucleic acid contains said modification. Alternatively, a nucleic acid encoding a modified Clostridial neurotoxin (or a part thereof) containing said modification can be synthesized.

[0291] Where the polypeptide sequence of a modified BoNT / A described herein includes a tag, such as a His tag, for example for purification, the tag is optional. Preferably, the tag is removed prior to use of the modified BoNT / A in the present invention.

[0292] As mentioned above, the modified BoNT / A described herein has increased tissue retention properties that also result in increased efficacy and / or duration of action, allowing for increased dosage without additional negative effects. One way in which these advantageous properties may be defined is in terms of the safety factor of the modified BoNT / A. In this regard, the undesirable effects of a clostridial toxin (caused by diffusion of the toxin from the site of administration) can be experimentally evaluated by measuring the rate of weight loss in a suitable animal model (e.g., mice in which weight loss is detected within 7 days of administration). Conversely, the desirable on-target effects of a clostridial toxin can be experimentally evaluated by the digit abduction score (DAS) assay, which is an index of muscle paralysis. The DAS assay may be performed by injecting 20 μl of a clostridial toxin formulated in gelatin phosphate buffer into the gastrocnemius / soleus muscle complex of mice, followed by evaluation of the digit abduction score using the method of Aoki (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 clostridial toxin injection, the varying degrees of digit abduction are scored on a 5-point scale (0=normal, 4=maximal reduction in digit abduction and limb extension).

[0293] The safety factor of a neurotoxin (e.g., a modified BoNT / A of the invention (or unmodified BoNT / A for comparison)) may then be expressed as the ratio of the amount of toxin required to achieve a 10% reduction in body weight (measured at peak effect in the first 7 days after administration to mice) to the amount of toxin required for a DAS score of 2. Thus, a high safety factor score is desirable and indicates a neurotoxin that can effectively paralyze a target muscle with few undesirable off-target effects. The modified BoNT / A of the invention has a higher safety factor than the safety factor of a comparable unmodified (native) BoNT / A.

[0294] A high safety margin is particularly advantageous in therapy, since it means an increased therapeutic index. In other words, this means that lower doses can be used and / or higher doses can be used without additional (e.g., harmful) effects, as compared to alternative Clostridial neurotoxin therapies. The possibility of using higher doses of a neurotoxin without additional effects is particularly advantageous, since higher doses usually increase the duration of action of the neurotoxin.

[0295] The efficacy of modified BoNT / A is evaluated by measuring the efficacy of a given DAS score, e.g., a DAS score of 2 (ED 50 The efficacy of modified BoNT / A may be expressed as the EC value (EC20) in a cellular assay measuring SNARE cleavage by the neurotoxin, or the minimum dose of neurotoxin that results in a DAS score of 4. 50 Dose, e.g., EC in a cellular assay measuring SNAP25 cleavage by modified BoNT / A 50 It may also be expressed as dose.

[0296] The duration of action of a modified BoNT / A may be expressed as the time required to restore a DAS score to 0 after administration of a given dose of neurotoxin, e.g., the minimum dose of neurotoxin that results in a DAS score of 4, to the gastrocnemius / soleus muscle complex of a mouse.

[0297] Thus, in one embodiment, a modified BoNT / A of the invention has a safety factor of greater than 7 (e.g., at least 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50), where the safety factor is calculated as: DAS ED 50 Dose of toxin required for 10% body weight change (pg / mouse) divided by ED 50 = dose required to produce a DAS score of 2. For example, the modified BoNT / A may have a safety margin of at least 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, or 50.

[0298] In one embodiment, a modified BoNT / A of the present invention has a safety factor of at least 10. In one embodiment, a modified BoNT / A of the present invention has a safety factor of at least 15. Preferably, the modified BoNT / A 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).

[0299] The modified BoNT / A 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: 3 to 7. For example, it is a polypeptide sequence having at least 80%, 90%, 95%, or 99.9% sequence identity to a polypeptide sequence selected from SEQ ID NOs: 3 to 7. Preferably, the modified BoNT / A used in the present invention may comprise a polypeptide sequence selected from SEQ ID NOs: 3 to 7 (more preferably, consists of said polypeptide sequence). Of the modified BoNT / As, SEQ ID NO: 6 is preferred.

[0300] Thus, it is desirable that the modified BoNT / A comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 6. More preferably, the polypeptide sequence has at least 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 6. Most preferably, the modified BoNT / A used in the present invention may comprise (and more preferably consist of) SEQ ID NO: 6.

[0301] The two-chain modified BoNT / A 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: 3 to 7 constituting the first chain of the two-chain modified BoNT / A, 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: 3 to 7 constituting the second chain of the two-chain modified BoNT / A. N and H Cdomain, where the first chain and the second chain are linked by a disulfide bond.

[0302] If cleavage occurs at one or more positions within the activation loop of a modified BoNT / A that includes a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 3-7, 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: 3-7 may be absent from the two-chain modified BoNT / A. With this in mind, the sequence of the two-chain modified BoNT / A (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: 3-7) may differ slightly from the sequence of the corresponding single-chain modified BoNT / A comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to any one of SEQ ID NOs: 3-7. The subfragment may be 1 to 15 amino acids. In particular, in one embodiment, when Lys-C is used to convert a single-chain modified BoNT / A to a two-chain modified BoNT / A, the subfragment of the C-terminal L chain portion of the absent sequence may be SEQ ID NO: 9 or 10.

[0303] Preferably, the two-chain modified BoNT / A 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: 6 constituting the first chain of the two-chain modified BoNT / A, 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: 6 constituting the second chain of the two-chain modified BoNT / A. N and H C domain, where the first and second chains are linked by a disulfide bond.

[0304] If cleavage occurs at one or more positions (preferably two positions) within the activation loop of a modified BoNT / A that includes a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 6, 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: 6 may be absent from the two-chain modified BoNT / A. With this in mind, the sequence of the two-chain modified BoNT / A (e.g., including a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 6) may differ slightly from the sequence of a corresponding single-chain modified BoNT / A that includes a polypeptide sequence having at least 70%, 80%, 90%, 95%, 99.9%, or 100% sequence identity to SEQ ID NO: 6. The subfragment can be 1 to 15 amino acids. In particular, in one embodiment, when Lys-C is used to convert a single-chain modified BoNT / A to a two-chain modified BoNT / A, the subfragment of the C-terminal L-chain portion of the absent sequence can be SEQ ID NO: 9 or 10.

[0305] In a particularly preferred embodiment, the two-chain modified BoNT / A comprises a light chain having a polypeptide sequence having at least 70%, 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 11 or 12 (preferably SEQ ID NO: 11) and a heavy chain comprising a polypeptide sequence having at least 70%, 80%, 90%, 95%, or 99.9% sequence identity to SEQ ID NO: 13, wherein said light chain and heavy chain are linked together by a disulfide bond. More preferably, the two-chain modified BoNT / A comprises (or consists of) a light chain having SEQ ID NO: 11 or 12 (preferably SEQ ID NO: 11) and a heavy chain having SEQ ID NO: 13, wherein said light chain and heavy chain are linked together by a disulfide bond. Even more preferably, the single-chain modified BoNT / A comprises (or consists of) a light chain comprising SEQ ID NO: 11 and a heavy chain comprising SEQ ID NO: 13, wherein said light chain and heavy chain are linked together by a disulfide bond. Said disulfide bond is preferably formed by and / or between cysteine ​​residue 429 of SEQ ID NO:11 or 12 and cysteine ​​residue 6 of SEQ ID NO:13.

[0306] In a preferred embodiment, the modified BoNT / A of the invention does not include a therapeutic or diagnostic agent (e.g., a nucleic acid, protein, peptide, or small molecule therapeutic or diagnostic agent) added to the light and heavy chains. For example, in one embodiment, the modified BoNT / A may not include a covalently or non-covalently bound therapeutic or diagnostic agent. Thus, the modified BoNT / A of the invention preferably does not function as a delivery vehicle for additional therapeutic or diagnostic agents.

[0307] When the modified BoNT / A described herein includes a tag (e.g., a His tag) and / or a linker, e.g., for purification, the tag and / or linker are optional. Preferably, the tag is removed prior to use of the modified BoNT / A in the present invention.

[0308] The modified BoNT / A is preferably uncomplexed (i.e., it may not contain complex proteins present in a naturally occurring Clostridial neurotoxin complex, e.g., a BoNT / A complex). Examples of such complex proteins include neurotoxin-associated proteins (NAPs) and non-toxic non-hemagglutinin components (NTNHs). However, it is preferred that the modified BoNT / A is a recombinant modified BoNT / A. The modified BoNT / A of the present invention can be produced using recombinant nucleic acid technology. Thus, in one embodiment, the modified BoNT / A (described herein) is a recombinant modified BoNT / A.

[0309] In one embodiment, a nucleic acid (e.g., DNA) is provided that comprises a nucleic acid sequence encoding a modified BoNT / A. 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 may be selected from any of the nucleic acid sequences described herein.

[0310] In a preferred embodiment, the vector comprises a promoter selected from: Promoter / Inducer / Typical induction conditions Tac (hybrid) / IPTG / 0.2mM (0.05-2.0mM) AraBAD / L-arabinose / 0.2% (0.002-0.4%) T7-lac operator / IPTG / 0.2 mM (0.05-2.0mM)

[0311] In another preferred embodiment, the vector comprises a promoter selected from: Promoter / Inducer / Typical induction conditions Tac (hybrid) / IPTG / 0.2mM (0.05-2.0mM) AraBAD / L-arabinose / 0.2% (0.002-0.4%) T7-lac operator / IPTG / 0.2 mM (0.05-2.0mM) T5-lac operator / IPTG / 0.2 mM (0.05-2.0mM)

[0312] The nucleic acid molecules may be produced using any suitable procedure known in the art. Thus, the nucleic acid molecules may be produced using chemical synthesis techniques. Alternatively, the nucleic acid molecules of the present invention may be produced using molecular biology techniques.

[0313] The DNA constructs of the present invention are preferably designed in silico and then synthesized by conventional DNA synthesis techniques.

[0314] The above nucleic acid sequence information is optionally modified for codon bias in the final host cell (eg, E. coli) expression system used.

[0315] The terms "nucleotide sequence" and "nucleic acid" are used interchangeably herein. Preferably, the nucleotide sequence is a DNA sequence.

[0316] The modified BoNT / A of the present invention may exist as a single chain or two chains, however, it is preferred that the modified BoNT / A exists as two chains in which the L chain is linked to the H chain (or a component thereof, e.g., the HN domain) via a disulfide bond.

[0317] The production of a single-chain modified BoNT / A having a light chain and a heavy chain may be achieved using a method comprising expressing a nucleic acid encoding the modified BoNT / A in an expression host, disrupting the host cells to provide a host cell homogenate containing the single-chain modified BoNT / A, and isolating the single-chain modified BoNT / A. The single-chain modified BoNT / A described herein may be proteolytically processed using a method comprising contacting the single-chain modified BoNT / A with a protease (e.g., Lys-C) that hydrolyzes a peptide bond in the activation loop of the modified BoNT / A, thereby converting the single-chain modified BoNT / A into the corresponding two-chain modified BoNT / A (e.g., where the light chain and the heavy chain are linked by a disulfide bond). The two-chain modified BoNT / A is preferably obtained by such a method.

[0318] Thus, the modified BoNT / A used in the present invention is preferably a two-chain modified BoNT / A 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 modified BoNT / A used in the present invention is preferably a two-chain modified BoNT / A 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:6. Most preferably, the modified BoNT / A used in the present invention is a two-chain modified BoNT / A generated from a polypeptide comprising SEQ ID NO:6 (and even more preferably consisting of SEQ ID NO:6). Thus, in some embodiments, the modified BoNT / A is a two-chain modified BoNT / A 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 modified BoNT / A comprising SEQ ID NO:6 with a protease that hydrolyzes a peptide bond in the activation loop, thereby converting the single-chain modified BoNT / A into the corresponding two-chain modified BoNT / A. In some embodiments, the modified BoNT / A is a two-chain modified BoNT / A in which the L chain is linked to the H chain via a disulfide bond, obtained by a method comprising contacting a single-chain modified BoNT / A consisting of SEQ ID NO:6 with a protease that hydrolyzes a peptide bond in the activation loop, thereby converting the single-chain modified BoNT / A into the corresponding two-chain modified BoNT / A.

[0319] 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 WO2014 / 080206, WO2014 / 079495, and EP2677029A2, which are incorporated herein by reference. Lys-C may cleave the activation loop C-terminus to one or more lysine residues present at the activation loop C-terminus. It will be understood by those skilled in the art that if Lys-C cleaves the activation loop multiple times, the small peptides of the activation loop of the dichain modified BoNT / A may not exist when compared to the sequence numbers shown herein.

[0320] As used herein, the term "obtained" encompasses the term "obtained." In one embodiment, the term "obtained" means obtained.

[0321] As used herein, the term "one or more" means 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 refer to every member of said list. Similarly, as used herein, "at least one" 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 refer to every member of said list.

[0322] As used herein, the word "illness" encompasses "disorder." In one embodiment, the disorder is a disease.

[0323] The modified BoNT / A of the invention may be formulated in any suitable manner for administration to a patient, for example, as part of a pharmaceutical composition comprising the modified BoNT / A of the invention and a pharma- ceutically acceptable carrier, excipient, adjuvant, propellant, and / or salt.

[0324] Liquid dosage forms are typically prepared using the modified BoNT / A and a pyrogen-free sterile solvent. The modified BoNT / A may be dissolved or suspended in the solvent, depending on the solvent and concentration used. In preparing a solution, the modified BoNT / A may be dissolved in the solvent, the solution may be made isotonic by adding sodium chloride as necessary, sterilized by filtration through a sterile filter using aseptic techniques, and then filled into a suitable sterile bottle or ampoule and sealed. Alternatively, if the stability of the solution is sufficient, the solution in a 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.

[0325] Dry powders that are dissolved or suspended in a suitable solvent prior to use may be prepared by filling sterile containers with pre-sterilized ingredients using aseptic technique in a sterile area. Alternatively, the ingredients may be dissolved in suitable containers using aseptic technique in a sterile area. The product is then freeze-dried and the containers are aseptically sealed.

[0326] Parenterally administered suspensions suitable for the routes of administration described herein are prepared in substantially the same manner, except that the sterile components are suspended in a sterile vehicle instead of being dissolved, and sterilization cannot be achieved by filtration. The components may be isolated under aseptic conditions or may be sterilized after isolation, for example by gamma irradiation.

[0327] Advantageously, a suspending agent, such as polyvinylpyrrolidone, is included in the composition to facilitate uniform distribution of the ingredients.

[0328] Also provided is a unit dosage form of modified BoNT / A for use in treating blepharospasm, typical hemifacial spasm and / or atypical hemifacial spasm, said unit dosage form comprising: (a) at least 10 units (preferably between 10 and 333 units) of modified BoNT / A, where one unit is the amount of modified BoNT / A that corresponds to the calculated median lethal dose (LD50) in mice; or (b) at least 240 pg (preferably between 240 pg and 8000 pg) of modified BoNT / A; and (c) optionally, pharma- ceutically acceptable carriers, excipients, adjuvants, and / or salts; Here, the modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (Hc domain).

[0329] The unit dose may be 1,500 to 5,000 pg modified BoNT / A, preferably 2,000 to 4,500 pg modified BoNT / A. Examples of suitable unit doses include about 2,500 pg (e.g., 2,000 pg ± 10%) modified BoNT / A; and about 4,000 pg (e.g., 4,000 pg ± 10%) modified BoNT / A.

[0330] The unit dose may be between 62.4 and 208 U of modified BoNT / A, preferably between 83.2 and 187.2 U of modified BoNT / A. Examples of suitable unit doses include about 104 U (e.g., 104 U ± 10%) of modified BoNT / A; and about 166.4 U (e.g., 166.4 U ± 10%) of modified BoNT / A.

[0331] A unit dosage form for use in treating blepharospasm, typical hemifacial spasm and / or atypical hemifacial spasm may contain 10 Units to 333 Units of modified BoNT / A. The upper end of the range may be 300, 250, 200, 150, or 100 Units of modified BoNT / A, preferably where the upper limit is 250 Units. The lower end of the range may be 40, 45, 50, 60, 65, 70, 75, 80, 85, 90, or 100 Units, preferably 50 Units. Preferably, the unit dosage form may contain 42 Units to 300 Units of modified BoNT / A, such as 200 Units to 300 Units of modified BoNT / A.

[0332] A unit dosage form for use in treating blepharospasm, typical hemifacial spasm and / or atypical hemifacial spasm may contain 240 Units to 8,000 Units of modified BoNT / A. The upper end of the range may be 7,500, 6,500, 5,500, 4,500, 3,500, 2,500, 1,500 or 500 pg of modified BoNT / A, preferably, where the upper end is 5,500 pg. The lower end of the range may be 750, 850, 950, 1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500 or 5,000 pg of modified BoNT / A, preferably, 1,000 Units. Preferably, the unit dosage form contains between 2,000 pg and 7,000 pg of modified BoNT / A, for example between 4,000 pg and 6,000 pg of modified BoNT / A.

[0333] Another aspect of the invention provides a kit comprising: (a) a unit dosage form of a modified BoNT / A as described herein, the modified BoNT / A comprising a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain); and (b) instructions for using the unit dosage form in the treatment of blepharospasm; and (c) Optionally, a diluent.

[0334] Another aspect of the invention provides a kit comprising: (a) a unit dosage form of a modified BoNT / A as described herein, wherein the modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain); and (b) instructions for use of the unit dosage form in the treatment of typical hemifacial spasm; and (c) Optionally, a diluent.

[0335] Another aspect of the invention provides a kit comprising: (a) a unit dosage form of a modified BoNT / A as described herein, wherein the modified BoNT / A comprises a BoNT / A light chain and translocation domain, and a BoNT / B receptor binding domain (HC domain); and (b) instructions for use of the unit dosage form in the treatment of atypical hemifacial spasm; and (c) Optionally, a diluent.

[0336] The modified BoNT / A in the unit dosage form may comprise a polypeptide sequence having at least 70% sequence identity to SEQ ID NOs: 3-7. For example, a polypeptide sequence having at least 80%, 90%, 95% or 99.5% sequence identity to any one of SEQ ID NOs: 3-7.

[0337] Preferably, the modified BoNT / A in the unit dosage form comprises a polypeptide sequence having at least 70% sequence identity to SEQ ID NO: 6. For example, a polypeptide sequence having at least 80%, 90%, 95% or 99.5% sequence identity to SEQ ID NO: 6. Most preferably, the modified BoNT / A may comprise (and more preferably consist of) SEQ ID NO: 6.

[0338] The various therapeutic application embodiments of the present invention may be applied to the methods of the present invention and vice versa.

[0339] sequence homology

[0340] To determine percent identity, any of a variety of sequence alignment methods can be used, including but not limited to global methods, local methods, and hybrid methods such as segment approach methods. Protocols for determining percent identity are routine procedures within the scope of those skilled in the art. In global methods, the molecular sequences are aligned from beginning to end, and the optimal alignment is determined by summing the scores of each residue pair and applying gap penalties. Non-limiting methods include, for example, CLUSTALW, see, for example, Julie D. Thompson et al., CLUSTAL W: Improving the Sensitivity of Progressive Multiple Sequence Alignment Through Sequence Weighting, Position- Specific Gap Penalties and Weight Matrix Choice, 22(22) Nucleic Acids Research 4673-4680 (1994); iterative improvement methods, see, for example, Osamu Gotoh, Significant Improvement in Accuracy of Multiple Protein. Sequence Alignments by Iterative Refinement as Assessed by Reference to Structural Alignments, 264(4) J. MoI. Biol. 823-838 (1996). Local methods align sequences by identifying one or more conserved motifs shared by all input sequences.Non-limiting methods include, for example, Matchbox, see, e.g., Eric Depiereux and Ernest Feytmans, Match-Box: A Fundamentally New Algorithm for the Simultaneous Alignment of Several Protein Sequences, 8(5) CABIOS 501 -509 (1992); Gibbs Sampling, see, e.g., C. E. Lawrence et al., Detecting Subtle Sequence Signals: A Gibbs Sampling Strategy for Multiple Alignment, 262(5131) Science 208-214 (1993); Align-M, see, e.g., Ivo Van WaIIe et al., Align-M - A New Algorithm for Multiple Alignment of Highly Divergent Sequences, 20(9) Bioinformatics:1428-1435 (2004).

[0341] Thus, the percent sequence identity is determined by conventional methods. See, for example, Altschul et al, Bull. Math. Bio. 48: 603-16, 1986 and Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915-19, 1992. Briefly, as shown below (amino acids are indicated by standard single letter code), two amino acid sequences are aligned using a gap open penalty of 10, a gap extension penalty of 1, and the "blosum62" score matrix of Henikoff and Henikoff (ibid.) to optimize the alignment score. Preferably, this method is used to align a sequence with the subject sequence herein (e.g., SEQ ID NO:2) and define the numbering of amino acid positions, as described herein.

[0342] The "percent sequence identity" between two or more nucleic acid or amino acid sequences is a function of the number of identical positions shared by the sequences. Thus, the percent identity may be calculated as the number of identical nucleotides / amino acids divided by the total number of nucleotides / amino acids multiplied by 100. The calculation of percent sequence identity may also take into account the number of gaps and the length of each gap that needs to be inserted to optimize the alignment of two or more sequences. The comparison of sequences and the determination of percent identity between two or more sequences can be carried out using certain mathematical algorithms, such as BLAST, which are well known to those skilled in the art.

[0343] Alignment scores for determining sequence identity

number

[0344] The percent identity is calculated as follows: [total number of perfect matches × 100] / [length of the longer sequence + number of gaps inserted into the longer sequence to align the two sequences]

[0345] Substantially homologous polypeptides are considered to have one or more amino acid substitutions, deletions or additions. These changes are preferably of a relatively minor nature, i.e., conservative amino acid substitutions (see below) and other substitutions that do not significantly affect the folding or activity of the polypeptide: small deletions, typically from one to about 30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue, small linker peptides of up to about 20-25 residues, or affinity tags.

[0346] Conservative Amino Acid Substitutions Basicity: Arginine lysine Histidine Acidic: Glutamic acid Aspartic acid polarity: glutamine Asparagine Hydrophobicity: Leucine Isoleucine Ballin Aromatic: Phenylalanine Tryptophan Tyrosine small: glycine Alanine Serine Threonine Methionine

[0347] In addition to the 20 standard amino acids, non-standard amino acids (such as 4-hydroxyproline, 6-N-methyllysine, 2-aminoisobutyric acid, isovaline, and α-methylserine) may be substituted for amino acid residues in the polypeptides of the invention. A limited number of non-conserved amino acids, as well as amino acids that are not encoded by the genetic code and that do not occur naturally, may be substituted for polypeptide amino acid residues. The polypeptides of the invention may also include non-naturally occurring amino acid residues.

[0348] Non-naturally occurring amino acids include, but are not limited to, trans-3-methylproline, 2,4-methanoproline, cis-4-hydroxyproline, trans-4-hydroxyproline, N-methylglycine, allothreonine, methylthreonine, hydroxyethylcysteine, hydroxyethylhomocysteine, nitroglutamine, homoglutamine, pipecolic acid, tertroucine, norvaline, 2-azaphenylalanine, 3-azaphenylalanine, 4-azaphenylalanine, and 4-fluorophenylalanine. Various methods are known in the art for incorporating non-naturally occurring amino acid residues into proteins. For example, an in vitro system can be used in which nonsense mutations are suppressed using chemically aminoacylated suppressor tRNAs. Methods for synthesizing amino acids and aminoacylating tRNAs are known in the art. Transcription and translation of the plasmid containing the nonsense mutation is carried out in a cell-free system containing an E. coli S30 extract and commercially available enzymes and other reagents. The protein is purified by chromatography. See, e.g., Robertson et al., J. Am. Chem. Soc. 113:2722, 1991; Ellman et al., Methods Enzymol. 202:301, 1991; Chung et al., Science 259:806-9, 1993; and Chung et al., Proc. Natl. Acad. Sci. USA 90:10145-9, 1993. In the second method, translation is driven in Xenopus oocytes by microinjection of mutant mRNA and chemically aminoacylated suppressor tRNA (Turcatti et al., J. Biol. Chem. 271:1991-8, 1996). In the third method, E. coli cells are cultured in the absence of the natural amino acid to be substituted (e.g., phenylalanine) and in the presence of the desired non-naturally occurring amino acid (e.g., 2-azaphenylalanine, 3-azaphenylalanine, 4-azaphenylalanine, or 4-fluorophenylalanine) that is incorporated into the polypeptide in place of the natural amino acid.See Koide et al., Biochem. 33:7470-6, 1994. Naturally occurring amino acid residues can be converted to non-naturally occurring species by in vitro chemical modification. Chemical modification can be combined with site-directed mutagenesis to further expand the range of such substitutions (Wynn and Richards, Protein Sci. 2:395-403, 1993).

[0349] A limited number of non-conservative amino acids, amino acids that are not encoded by the genetic code, non-naturally occurring amino acids, and unnatural amino acids may be substituted for amino acid residues in the polypeptides of the invention.

[0350] Essential amino acids in the polypeptides of the invention can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis (Cunningham and Wells, Science 244: 1081-5, 1989). Biological interaction sites can also be determined by physical analysis of structures determined by techniques such as nuclear magnetic resonance, crystallography, electron diffraction or photoaffinity labeling, in combination with mutations of putative contact site amino acids. See, for example, de Vos et al., Science 255:306-12, 1992; Smith et al., J. Mol. Biol. 224:899-904, 1992; Wlodaver et al., FEBS Lett. 309:59-64, 1992. The identity of essential amino acids can also be inferred from analysis of homology with related components of the polypeptides of the invention, such as translocation or protease components.

[0351] Multiple amino acid substitutions can be made and tested using known methods for mutagenesis and screening, such as those disclosed by Reidhaar-Olson and Sauer (Science 241:53-7, 1988) or Bowie and Sauer (Proc. Natl. Acad. Sci. USA 86:2152-6, 1989). Briefly, these authors disclose methods for simultaneously randomizing two or more positions within a polypeptide, selecting functional polypeptides, and then sequencing the mutagenized polypeptides to determine the spectrum of substitutions tolerated at each position. Other methods that can be used include phage display (e.g., Lowman et al., Biochem. 30:10832-7, 1991; Ladner et al., US Patent No. 5,223,409; Huse, WIPO Publication WO 92 / 06204) and site-directed mutagenesis (Derbyshire et al., Gene 46:145, 1986; Ner et al., DNA 7:127, 1988).

[0352] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Singleton, et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 20 ED., John Wiley and Sons, New York (1994), and Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY, Harper Perennial, NY (1991) provide those of ordinary skill in the art with a general dictionary of many of the terms used in this disclosure.

[0353] The disclosure is not limited by the exemplary methods and materials disclosed herein, and any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the disclosure. Numeric ranges include the numbers defining the range. Unless otherwise specified, any nucleic acid sequence is written left to right in a 5' to 3' orientation, and any amino acid sequence is written left to right in an amino to carboxy orientation, respectively.

[0354] The headings provided herein are not intended to limit the various aspects or embodiments of the disclosure.

[0355] Amino acids are referred to herein using the amino acid name, three-letter abbreviation, or one-letter abbreviation. As used herein, the term "protein" includes proteins, polypeptides, and peptides. As used herein, the term "amino acid sequence" is synonymous with the term "polypeptide" and / or the term "protein." In some cases, the term "amino acid sequence" is synonymous with the term "peptide." In some cases, the term "amino acid sequence" is synonymous with the term "enzyme." The terms "protein" and "polypeptide" are used interchangeably herein. In the present disclosure and claims, conventional single-letter and three-letter codes for amino acid residues may be used. The three-letter codes for amino acids are as defined in accordance with the IUPACIUB Joint Commission on Biochemical Nomenclature (JCBN). It is also understood that due to the degeneracy of the genetic code, a polypeptide may be encoded by one or more nucleotide sequences.

[0356] Different definitions of terms may appear throughout the specification. Before a more detailed description of the exemplary embodiments, it is to be understood that the present disclosure is not limited to the particular embodiments described, as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure is defined only by the appended claims.

[0357] Where a range of values ​​is given, it is understood that each intervening value between the upper and lower limits of said range is also expressly disclosed...

Claims

1. A pharmaceutical agent for use in a method for treating blepharospasm in a patient, comprising a modified double-chain chimeric botulinum neurotoxin BoNT / AB, wherein the BoNT / AB is administered by intramuscular injection to multiple sites on the patient's face, and the method comprises: a) Administer BoNT / AB at a dose of 1 unit per injection site to up to six different injection sites in the orbicularis oculi superioris muscle proximal to the first eye of the patient, where the up to six different injection sites are selected from the following: i) medial superior orbital septum orbicularis oculi muscle; ii) superior orbital orbicularis oculi; iii) lateral superior orbital septum anterior orbicularis oculi muscle; iv) lateral orbital orbicularis; v) Anterior part of the medial superior tarsal plate, orbicularis oculi muscle; and vi) The orbicularis oculi muscle of the anterior part of the lateral superior tarsal plate; and / or b) Administer BoNT / AB at a dose of 1 unit per injection site to up to two different injection sites in the orbicularis oculi muscle proximal to the first eye of the patient, wherein the up to two different injection sites are selected from the following: i) Orbicularis oculi muscle medialis; and ii) Lateral orbicularis oculi muscle; and / or c) Administer BoNT / AB at a dose of 1 unit per injection site to up to two different injection sites selected from the following: i) Two different injection sites in the corrugator supercilii muscle proximal to the first eye of the patient; and ii) One site on the procerus muscle proximal to the first eye of the patient; Here, the unit dose of BoNT / AB is at least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of BoNT / AB, and the BoNT / AB comprises the BoNT / A light chain and the translocation domain (H N The domain) and the BoNT / B receptor binding domain (H C Domain) and, including

2. The pharmaceutical product according to claim 1, wherein BoNT / AB is administered by intramuscular injection to at least six different injection sites on the patient's face.

3. The pharmaceutical product according to claim 1 or 2. The method for treating blepharospasm further comprises: (a) Administer BoNT / AB at a dose of 1 unit per injection site to up to six different injection sites in the orbicularis oculi muscle proximal to the second eye of the patient, wherein the up to six different injection sites are selected from the following: (i) Medial superior orbital septum anterior orbicularis oculi muscle; (ii) superior orbital orbicularis oculi; (iii) lateral superior orbital septum anterior orbicularis oculi muscle; (iv) lateral orbital orbicularis oculi; (v) Anterior part of the medial superior tarsal plate, orbicularis oculi muscle; and (vi) The orbicularis oculi muscle of the anterior part of the lateral superior tarsal plate; and / or (b) Administer BoNT / AB at a dose of 1 unit per injection site to up to two different injection sites in the orbicularis oculi inferior muscle proximal to the second eye of the patient, wherein the up to two different injection sites are selected from the following: (i) medial orbicularis oculi muscle; and (ii) Lateral orbicularis oculi muscle; and / or (c) Administer BoNT / AB at a dose of 1 unit per injection site to up to two different injection sites selected from the following: (i) Two different injection sites in the corrugator supercilii muscle proximal to the second eye of the patient; and (ii) One site on the procerus muscle proximal to the second eye of the aforementioned patient

4. The pharmaceutical product according to claim 3, wherein BoNT / AB is administered by intramuscular injection to at least six different sites in each of the patient's eyes.

5. The pharmaceutical product according to claim 1 or 2, wherein the method includes: (a) Administer one unit dose of BoNT / AB to the orbicularis oculi muscle anterior to the lateral superior orbital septum proximal to the first eye of the patient; (b) Administering one unit dose of BoNT / AB to the anterior orbicularis oculi muscle of the medial superior orbital septum proximal to the first eye of the patient; and (c) Administering one unit dose of BoNT / AB to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) located proximal to the first eye of the patient.

6. A pharmaceutical agent for use in a method for treating typical hemifacial spasm, comprising a modified double-chain chimeric botulinum neurotoxin BoNT / AB, wherein the BoNT / AB is administered by intramuscular injection to multiple sites on the face of the patient, and the method comprises: (a) Administer BoNT / AB at a dose of 1 unit per injection site to up to six different injection sites in the orbicularis oculi superioris muscle proximal to the first eye of the patient, wherein the up to six different injection sites are selected from the following: (i) Medial superior orbital septum anterior orbicularis oculi muscle; (ii) superior orbital orbicularis oculi; (iii) lateral superior orbital septum anterior orbicularis oculi muscle; (iv) lateral orbital orbicularis oculi; (v) Anterior part of the medial superior tarsal plate, orbicularis oculi muscle; and (vi) The orbicularis oculi muscle of the anterior part of the lateral superior tarsal plate; and / or (b) Administer BoNT / AB at a dose of 1 unit per injection site to up to two different injection sites in the orbicularis oculi inferior muscle proximal to the first eye of the patient, wherein the up to two different injection sites are selected from the following: (i) medial orbicularis oculi muscle; and (ii) Lateral orbicularis oculi muscle; and / or (c) Administer BoNT / AB at a dose of 1 unit per injection site to up to two different injection sites selected from the following: (i) Two different injection sites in the corrugator supercilii muscle proximal to the first eye of the patient; and (ii) One site on the procerus muscle proximal to the first eye of the patient; and / or (d) Administer one or more unit doses of BoNT / AB to one or more additional muscles affected by the hemifacial spasm, according to the following administration schedule: (i) One unit dose to the orbicularis oris superioris muscle; (ii) One unit dose to the orbicularis oris inferior muscle; (iii) One unit dose to the zygomaticus major muscle; (iv) One unit dose to the zygomaticus minor muscle; (v) A maximum dose of 5 units (preferably 1 unit) is administered to the frontalis muscle; (vi) One unit dose to the mentalis muscle; (vii) One unit dose to the platysma muscle; (viii) A maximum dose of 2 units (preferably 1 unit) to the corrugator supercilii muscle; (ix) One unit dose to the buccinator muscle; (x) A maximum dose of 2 units (preferably 1 unit) is administered to the masseter muscle; (xi) One unit dose to the procerus; (xii) One unit dose to the bridge of the nose; and / or (xiii) One unit dose to the levator palpebrae superioris muscle; Here, the unit dose of BoNT / AB is at least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of BoNT / AB, and the BoNT / AB is the light chain and translocation domain (H) of the botulinum neurotoxin BoNT / A. N The domain) and the BoNT / B receptor binding domain (H C Domain) and, including

7. The pharmaceutical product according to claim 6, wherein the BoNT / AB is administered by intramuscular injection to at least six different sites in the patient.

8. The pharmaceutical product according to claim 6 or 7, wherein the method includes: (a) Administer one unit dose of BoNT / AB to the orbicularis oculi muscle anterior to the lateral superior orbital septum proximal to the first eye of the patient; (b) Administering one unit dose of BoNT / AB to the anterior orbicularis oculi muscle of the medial superior orbital septum proximal to the first eye of the patient; and (c) Administering one unit dose of BoNT / AB to the lateral inferior orbicularis oculi muscle (preferably the lateral inferior orbital septum anterior orbicularis oculi muscle) located proximal to the first eye of the patient.

9. A pharmaceutical preparation for use in the treatment of atypical hemifacial spasm, comprising a modified double-chain chimeric botulinum neurotoxin BoNT / AB, wherein the BoNT / AB is administered by intramuscular injection to multiple sites on the face of the patient, and the method comprises: a) Administering a 1-unit dose of BoNT / AB to the orbicularis oculi muscle affected by hemifacial spasm (for example, administering a 1-unit dose of BoNT / AB to the superior orbicularis oculi muscle affected by hemifacial spasm, and / or administering a 1-unit dose of BoNT / AB to the inferior orbicularis oculi muscle affected by hemifacial spasm; preferably, administering a 1-unit dose of BoNT / AB to the superior orbicularis oculi muscle affected by hemifacial spasm, and a 1-unit dose of BoNT / AB to the inferior orbicularis oculi muscle affected by hemifacial spasm); and b) Optionally, administer one or more units of BoNT / AB to one or more additional muscles affected by hemifacial spasm according to the following administration plan: (i) One unit dose to the zygomaticus major muscle; (ii) One unit dose to the zygomaticus minor muscle; (iii) A maximum dose of 5 units (preferably 1 unit) is administered to the frontalis muscle; (iv) One unit dose to the mentalis muscle; (v) One unit dose to the platysma muscle; (vi) One unit dose to the buccinator muscle; (vii) A maximum dose of 2 units (preferably 1 unit) is administered to the masseter muscle; (viii) One unit dose applied to the bridge of the nose; (ix) One unit dose to the levator palpebrae superioris muscle; and / or c) Optionally, administer one or more units of BoNT / AB to one or more additional muscles affected by hemifacial spasm according to the following administration plan: One unit dose per injection site, administered to up to six different injection sites in the orbicularis oculi muscle selected from the following: (i) Medial superior orbital septum anterior orbicularis oculi muscle; (ii) superior orbital orbicularis oculi; (iii) lateral superior orbital septum anterior orbicularis oculi muscle; (iv) lateral orbital orbicularis oculi; (v) Anterior part of the medial superior tarsal plate, orbicularis oculi muscle; and (vi) The orbicularis oculi muscle of the anterior part of the lateral superior tarsal plate; and / or One unit dose per injection site, administered to up to two different injection sites in the orbicularis oculi muscle selected from the following: (i) medial orbicularis oculi muscle; and (ii) Lateral orbicularis oculi muscle; and / or One unit dose per injection site, to a maximum of two different injection sites selected from the following: (i) Two different injection sites of the corrugator supercilii muscle; and (ii) One site on the procerus muscle; Here, the unit dose of BoNT / AB is at least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of BoNT / AB, and the BoNT / AB consists of botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N ) and the BoNT / B receptor binding domain (H C Domain) and, including

10. A pharmaceutical agent for use in a method for treating blepharospasm in a patient, comprising a modified double-chain chimeric botulinum neurotoxin BoNT / AB, wherein the BoNT / AB is administered by intramuscular injection to multiple sites on the patient's face, and the method comprises: a) Administer one unit dose of BoNT / AB to the lateral orbicularis superior oculi muscle proximal to the first eye of the patient; b) Administering one unit dose of BoNT / AB to the medial orbicularis superior oculi muscle proximal to the first eye of the patient; and c) Administer a 1-unit dose of BoNT / AB to the lateral orbicularis oculi muscle located proximal to the first eye of the patient. Here, the unit dose of BoNT / AB is at least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of BoNT / AB, and the BoNT / AB consists of the BoNT / A light chain and the translocation domain (H N ) and the BoNT / B receptor binding domain (H C Domain) and, including

11. A pharmaceutical agent for use in a method for treating typical hemifacial spasm, comprising a modified double-chain chimeric botulinum neurotoxin BoNT / AB. Here, BoNT / AB is administered by intramuscular injection to multiple sites on the patient's face, and the method includes the following: a) Administer one unit dose of BoNT / AB to the lateral superior orbicularis oculi muscle proximal to the eye affected by hemifacial spasm; b) Administer one unit dose of BoNT / AB to the medial superior orbicularis oculi muscle proximal to the eye affected by hemifacial spasm; c) Administering one unit dose of BoNT / AB to the lateral inferior orbicularis oculi muscle proximal to the eye affected by hemifacial spasm; and d) Administer one or more unit doses of BoNT / AB to one or more additional muscles affected by hemifacial spasm according to the following administration plan: (i) One unit dose to the orbicularis oris superioris muscle; (ii) One unit dose to the orbicularis oris inferior muscle; (iii) One unit dose to the zygomaticus major muscle; (iv) One unit dose to the zygomaticus minor muscle; (v) A maximum dose of 5 units (preferably 1 unit) is administered to the frontalis muscle; (vi) One unit dose to the mentalis muscle; (vii) One unit dose to the platysma muscle; (viii) A maximum dose of 2 units (preferably 1 unit) to the corrugator supercilii muscle; (ix) One unit dose to the buccinator muscle; (x) A maximum dose of 2 units (preferably 1 unit) is administered to the masseter muscle; (xi) One unit dose to the procerus; (xii) One unit dose to the bridge of the nose; and / or (xiii) One unit dose to the levator palpebrae superioris muscle; Here, the unit dose of the BoNT / AB is at least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of BoNT / AB, and the BoNT / AB comprises a botulinum neurotoxin A (BoNT / A) light chain and translocation domain (H N ), and a BoNT / B receptor binding domain (H C domain).

12. A pharmaceutical agent for use in a method for treating atypical hemifacial spasm, comprising a modified double-chain chimeric botulinum neurotoxin BoNT / AB, wherein the BoNT / AB is injected intramuscularly into multiple sites on the face of the patient, and the method comprises: a) Administering a 1-unit dose of BoNT / AB to the orbicularis oculi muscle affected by hemifacial spasm (for example, administering a 1-unit dose of BoNT / AB to the superior orbicularis oculi muscle affected by hemifacial spasm, and / or administering a 1-unit dose of BoNT / AB to the inferior orbicularis oculi muscle affected by hemifacial spasm; preferably, administering a 1-unit dose of BoNT / AB to the superior orbicularis oculi muscle affected by hemifacial spasm, and a 1-unit dose of BoNT / AB to the inferior orbicularis oculi muscle affected by hemifacial spasm); and b) Optionally, administer one or more units of BoNT / AB to one or more additional muscles affected by hemifacial spasm according to the following administration plan: (i) One unit dose to the zygomaticus major muscle; (ii) One unit dose to the zygomaticus minor muscle; (iii) A maximum dose of 5 units (preferably 1 unit) is administered to the frontalis muscle; (iv) One unit dose to the mentalis muscle; (v) One unit dose to the platysma muscle; (vi) A maximum dose of 2 units (preferably 1 unit) to the corrugator supercilii muscle; (vii) One unit dose to the buccinator muscle; (viii) A maximum dose of 2 units (preferably 1 unit) to the masseter muscle; (ix) One unit dose to the procerus; (x) One unit dose applied to the bridge of the nose; (xi) One unit dose administered to the lateral orbicularis oculi muscle; (xii) One unit dose administered to the medial superior orbicularis oculi muscle; (xiii) One unit dose administered to the lateral orbicularis oculi muscle; and / or (xiv) One unit dose to the levator palpebrae superioris muscle; Here, the unit dose of BoNT / AB is at least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB, the total dose administered during treatment is greater than 24,000 pg and up to 82,500 pg (e.g., up to 75,000 pg) of BoNT / AB, and the BoNT / AB is the light chain and translocation domain (H) of the botulinum neurotoxin BoNT / A. N ) and the BoNT / B receptor binding domain (H C Domain) and, including

13. The pharmaceutical agent according to claim 1 or 10, wherein the method further comprises administering one or more unit doses of the BoNT / AB to one or more additional muscles affected by blepharospasm according to the following administration schedule. (i) One unit dose to the orbicularis oris superioris muscle; (ii) One unit dose to the orbicularis oris inferior muscle; (iii) One unit dose to the zygomaticus major muscle; (iv) One unit dose to the zygomaticus minor muscle; (v) A maximum dose of 5 units (preferably 1 unit) is administered to the frontalis muscle; (vi) One unit dose to the mentalis muscle; (vii) One unit dose to the platysma muscle; (viii) A maximum dose of 2 units (preferably 1 unit) to the corrugator supercilii muscle; (ix) One unit dose to the buccinator muscle; (x) A maximum dose of 2 units (preferably 1 unit) is administered to the masseter muscle; (xi) One unit dose to the procerus; (xii) One unit dose to the bridge of the nose; and / or (xiii) One unit dose to the levator palpebrae superioris muscle.

14. The pharmaceutical agent according to claim 1 or 10, wherein the method further comprises administering one or more unit doses of the BoNT / AB to one or more additional muscles affected by blepharospasm according to the following administration schedule. (i) One unit dose to the levator palpebrae superioris muscle.

15. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose of BoNT / AB is 240 pg to 5,500 pg.

16. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose of BoNT / AB is 240 pg to 5,000 pg.

17. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose of BoNT / AB is 240 pg to 2,400 pg.

18. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose of BoNT / AB is 240 pg to 2,000 pg.

19. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose (e.g., the lower limit of a single unit dose) is at least 500 pg BoNT / AB.

20. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose (e.g., the lower limit of a single unit dose) is at least 1,000 pg BoNT / AB.

21. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose of BoNT / AB is 2,000 pg to 4,500 pg.

22. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the total dose administered during treatment is 25,000 pg to 50,000 pg BoNT / AB.

23. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the unit dose of BoNT / AB is 2,000 pg to 4,500 pg of BoNT / AB, and the total dose administered during treatment is 25,000 pg to 50,000 pg of BoNT / AB.

24. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the BoNT / AB comprises a polypeptide sequence having at least 70% sequence identity with respect to SEQ ID NO:

6.

25. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein BoNT / AB has a safety factor greater than 7, and the safety factor is calculated as follows: the dose of toxin required to change the body weight measured in pg / mouse by -10% is equal to the DAS ED measured in pg / mouse. 50 Divide by and ED 50 This is the dose required to obtain a DAS score of 2.

26. The pharmaceutical agent according to claim 1 or 10, wherein the method for treating the blepharospasm further comprises: Administer one unit dose of BoNT / AB to the lateral orbicularis oculi muscle proximal to the second eye of the aforementioned patient; Administer one unit dose of BoNT / AB to the medial orbicularis superior oculi muscle proximal to the second eye of the aforementioned patient; and Administer one unit dose of BoNT / AB to the lateral orbicularis oculi muscle located proximal to the second eye of the aforementioned patient.

27. ​​The pharmaceutical agent according to claim 1 or 10. The method for treating the blepharospasm further comprises administering: (i) the medial orbicularis oculi muscle located proximal to the patient's eye, (ii) the frontalis muscle located proximal to the patient's eye, and / or (iii) the corrugator supercilii muscle located proximal to the patient's eye, one unit dose of the BoNT / AB

28. The pharmaceutical product according to claim 1, 2, 6, 7, 9, 10, 11, or 12, wherein the BoNT / AB is administered in a single unit dose per injection site.

29. A unit dosage form of the modified double-chain chimeric botulinum neurotoxin BoNT / AB. The unit dosage form includes the following: a) At least 10 units (preferably 10 to 333 units) of BoNT / AB, where 1 unit is the calculated median lethal dose (LD) of a mouse. 50 This is the amount of BoNT / AB equivalent to ); b) At least 240 pg (preferably 240 pg to 8,000 pg) of BoNT / AB; and c) Optionally, pharmaceutically acceptable carriers, excipients, adjuvants, and / or salts, wherein BoNT / AB comprises BoNT / A light chain and transposition domain and BoNT / B receptor-binding domain (H C Domain) and, including

30. The kit includes the following: (a) The unit dosage form according to claim 29; and (b) Instructions for the use of the drug in the treatment of blepharospasm; and (c) Diluent at your discretion

31. The kit includes the following: (a) The unit dosage form according to claim 29; and (b) Instructions for the use of the drug in the treatment of typical hemifacial spasm; and (c) Diluent at your discretion

32. The kit includes the following: (a) The unit dosage form according to claim 29; and (b) Instructions for the use of the drug in the treatment of atypical hemifacial spasm; and (c) Diluent at your discretion

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