Non-protein clostridial toxin compositions

Abstract

To provide pharmaceutical compositions that stabilize a clostridial toxin active component.SOLUTION: The composition may be a liquid or solid composition, and comprises a surfactant and an antioxidant. In some embodiments, the composition comprises a surfactant selected from poloxamer and polysorbate, an antioxidant selected from methionine, N-acetylcysteine, ethylenediamine tetraacetic acid and combinations thereof, optionally an isotonicity agent and / or a freeze-drying protective agent selected from trehalose and sucrose.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to solid and liquid pharmaceutical compositions comprising a clostridial toxin active ingredient and one or more non-protein excipients. [Background technology]

[0002] A pharmaceutical composition is a formulation that contains at least one active ingredient (such as a clostridial toxin) and, for example, one or more excipients, buffers, carriers, stabilizers, preservatives, and / or bulking agents, and is suitable for administration to a patient to achieve a desired diagnostic or therapeutic effect. The pharmaceutical compositions disclosed herein have diagnostic, therapeutic, and / or research utility.

[0003] For storage stability and ease of handling, pharmaceutical compositions can be formulated as lyophilized (i.e., freeze-dried), i.e., vacuum-dried, powders that can be reconstituted with a suitable fluid, such as saline or water, before administration to a patient. Alternatively, pharmaceutical compositions can be formulated as aqueous solutions or suspensions. Pharmaceutical compositions can contain protein active ingredients. Unfortunately, protein active ingredients are very difficult to stabilize (i.e., maintain in a state that minimizes loss of biological activity), which can result in loss of protein and / or protein activity during formulation, reconstitution (if necessary), and storage of the pharmaceutical composition prior to use. Stability problems can arise due to surface adsorption, physical instability (e.g., denaturation or aggregation), or chemical instability (e.g., cross-linking, deamidation, isomerization, oxidation, formation of acidic or basic species, Maillard reaction, cleavage, etc.) of the protein active ingredient. To prevent such instability, various protein-based excipients, such as albumin and gelatin, have been used to stabilize protein active ingredients present in pharmaceutical compositions.

[0004] Unfortunately, despite their known stabilizing effects, the use of protein excipients such as albumin or gelatin in pharmaceutical compositions has significant drawbacks. For example, albumin and gelatin are expensive and increasingly difficult to obtain. Furthermore, when administered to a patient, blood products or animal-derived products such as albumin and gelatin may expose the patient to the potential risk of exposure to blood pathogens or infectious agents. Therefore, it is known that the presence of animal-derived protein excipients in pharmaceutical compositions may result in the inadvertent incorporation of infectious elements into the pharmaceutical composition. For example, it has been reported that the use of human serum albumin can transmit prions to pharmaceutical compositions. Therefore, it is desirable to find suitable non-protein excipients, such as stabilizers, cryoprotectants, and lyoprotectants, that can be used to stabilize protein active ingredients present in pharmaceutical compositions.

[0005] The inherent properties of clostridial toxins further limit and hinder the selection of suitable non-protein excipients for pharmaceutical compositions containing clostridial toxin active ingredients. For example, clostridial toxins are large proteins with an average molecular weight of approximately 150 kDa, and they are further complexed with non-toxin-associated proteins that increase their size to approximately 300-900 kDa. The size of clostridial toxin complexes makes them much more fragile and unstable than smaller, less complexed proteins, thereby making formulation and handling more difficult when maintaining clostridial toxin stability. Therefore, the use of non-protein excipients, such as stabilizers, cryoprotectants, and lyoprotectants, must be able to interact with the clostridial toxin active ingredient in a manner that does not denature, cleave, or otherwise inactivate the toxin or cause dissociation of non-toxin-associated proteins present in the toxin complex.

[0006] Another issue associated with clostridial toxin active ingredients is the extraordinary safety, precision, and accuracy required at every step of the formulation process, so non-protein excipients must not themselves be toxic or difficult to handle, so as not to exacerbate these already very stringent requirements.

[0007] Yet another difficulty associated with clostridial toxin active ingredients is the extremely low amount of clostridial toxin used in pharmaceutical compositions. Like enzymes, the biological activity of clostridial toxins generally depends, at least in part, on their three-dimensional structure. Therefore, clostridial toxins are detoxified by heat, various chemicals, surface stretching, and surface drying. Additionally, dilution of clostridial toxin complexes obtained by known culture, fermentation, and purification methods to the much lower concentrations used in pharmaceutical compositions is known to result in rapid inactivation of the toxin. The very small amounts of clostridial toxin active ingredient used in pharmaceutical compositions make this active ingredient highly susceptible to adsorption to, for example, laboratory glassware and containers, vials into which the pharmaceutical composition is reconstituted, and the inner surfaces of syringes used to inject the pharmaceutical composition. Such adsorption of the clostridial toxin active ingredient to surfaces can result in loss of the active ingredient and denaturation of the remaining clostridial toxin active ingredient, both of which reduce the overall activity of the active ingredient present in the pharmaceutical composition. Thus, the use of non-protein excipients, such as, for example, stabilizers, cryoprotectants and lyoprotectants, must be able to act as surface blocking agents to prevent adsorption of the clostridial toxin active ingredient to surfaces.

[0008] Yet another problem associated with clostridial toxin active ingredients is the pH sensitivity associated with complex formation. For example, the 900 kDa BoNT / A complex is known to be soluble in dilute aqueous solutions at pHs between 3.5 and 6.8. However, at pHs above about 7, non-toxicity-associated proteins dissociate from the 150 kDa neurotoxin, resulting in a loss of toxicity, particularly as the pH increases above 8.0. See Edward J. Schantz et al., pp. 44-45, "Preparation and characterization of botulinum toxin type A for human treatment," in Jankovic, J., et al., "Therapy with Botulinum Toxin" (Marcel Dekker, Inc., 1994). Because non-toxicity-associated proteins are thought to preserve or help stabilize the secondary and tertiary structures on which toxicity depends, dissociation of these proteins results in a less stable clostridial toxin active ingredient. Therefore, non-protein excipients useful for formulating pharmaceutical compositions containing a clostridial toxin active ingredient must be capable of operating within the range of pH levels necessary to maintain the activity of the clostridial toxin active ingredient.

[0009] Therefore, what is needed is a clostridial toxin pharmaceutical composition in which a clostridial toxin active ingredient (such as a botulinum toxin) is stabilized by a non-protein excipient. The present invention relates to solid and liquid clostridial toxin pharmaceutical compositions having one or more non-protein excipients that function to stabilize a clostridial toxin active ingredient present in the solid or liquid pharmaceutical composition. Summary of the Invention

[0010] In one aspect, a pharmaceutical composition is provided comprising a clostridial toxin active ingredient, a tonicity agent, a surfactant, and an antioxidant. In some embodiments, the pharmaceutical composition comprises a botulinum toxin. In some embodiments, the pharmaceutical composition comprises trehalose. In some embodiments, the pharmaceutical composition comprises sodium chloride. In some embodiments, the composition comprises a poloxamer and / or a polysorbate. In some embodiments, the composition comprises poloxamer 188 and / or polysorbate 20. In some embodiments, the antioxidant is selected from the group consisting of L-methionine, N-acetyl-cysteine (NAC), butylated hydroxytoluene (BHT), ethylenediaminetetraacetic acid sodium salt (EDTA), EDTA analogs, ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), EGTA analogs, diethylenetriaminepentaacetic acid (DTPA), DTPA analogs, ascorbic acid, and combinations thereof. In some embodiments, the antioxidant is methionine. In some embodiments, the antioxidant is NAC. In some embodiments, the antioxidants are NAC and EDTA. In some embodiments, the composition further comprises a buffering agent. In one embodiment, the buffering agent comprises a histidine buffering agent. In some embodiments, the composition has a pH of 5-7. In some embodiments, the composition is a liquid formulation. In some embodiments, the composition is a solid formulation.

[0011] In one aspect, the disclosure provides a liquid pharmaceutical composition comprising a clostridial toxin active ingredient, trehalose, poloxamer 188 or polysorbate 20, and L-methionine or N-acetyl-cysteine (NAC). In some embodiments, the liquid pharmaceutical composition comprises a botulinum toxin. In some embodiments, the liquid pharmaceutical composition further comprises EDTA or an EDTA analog. In some embodiments, the liquid pharmaceutical composition comprises a histidine buffer. In some embodiments, the pH of the liquid pharmaceutical composition is within the range of 5 to 7. In some embodiments, the relative weight of L-methionine is within the range of about 0.1% to about 0.3%. In some embodiments, the relative weight of NAC is within the range of about 0.1% to about 0.5%. In some embodiments, the relative weight of EDTA is within the range of about 0.01% to about 0.05%. In some embodiments, the relative weight of trehalose is within the range of about 1.0% to about 10%. In some embodiments, the relative weight of poloxamer 188 is within the range of about 2% to about 5%. In some embodiments, the relative weight amount of polysorbate 20 is in the range of about 0.02% to about 0.06%.

[0012] In another aspect, the present disclosure provides a solid pharmaceutical composition comprising a botulinum toxin, trehalose, poloxamer 188 or polysorbate 20, NAC, and EDTA or an EDTA analog. In an alternative embodiment, the solid pharmaceutical composition comprises a botulinum toxin, trehalose, poloxamer 188, and NAC. In an alternative embodiment, the solid pharmaceutical composition comprises a botulinum toxin, trehalose, poloxamer 188, and L-methionine. In some embodiments, the solid pharmaceutical composition further comprises a histidine buffer. In some embodiments, the relative weight amount of L-methionine is in the range of about 0.1% to about 0.3%. In some embodiments, the relative weight amount of NAC is in the range of about 0.01% to about 0.05%. In some embodiments, the relative weight amount of EDTA is in the range of about 0.01% to about 0.05%. In some embodiments, the relative weight amount of trehalose is in the range of about 1.0% to about 10%. In some embodiments, the relative weight amount of poloxamer 188 is in the range of about 0.5% to about 5%. In some embodiments, the relative weight amount of polysorbate 20 is in the range of about 0.02% to about 0.06%. DETAILED DESCRIPTION OF THE INVENTION

[0013] Certain compositions of the invention provide stable liquid or solid pharmaceutical compositions comprising a clostridial toxin active ingredient, a disaccharide, a surfactant, and an antioxidant. In certain liquid compositions of the invention, the disaccharide is optional.

[0014] Certain embodiments also provide methods of treating various diseases, disorders, and conditions using compositions provided according to aspects of the present invention, such as, for example, depression (e.g., major depressive disorder), headaches (e.g., migraines, tension headaches, etc.), pain, atrial fibrillation, hyperhidrosis, muscle spasticity, cervical dystonia, blepharospasm, overactive bladder (e.g., neurogenic detrusor overactivity and idiopathic overactive bladder), bladder pain (e.g., interstitial cystitis, painful bladder syndrome), skin conditions (e.g., wrinkles, fine lines, excessive sebum production, acne, rosacea), constipation, etc. Embodiments can include various administration techniques, including, for example, injection, such as intramuscular, intradermal, subcutaneous, ophthalmic, intravenous, transdermal, and topical. definition

[0015] As used herein, the words or terms set forth below have the following definitions:

[0016] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0017] As used herein, "about" or "approximately" means within an acceptable range of error for a particular value, as judged by one of ordinary skill in the art, and will depend in part on how the value is measured or determined (i.e., the limitations of the measurement system). For example, "about" can mean within 1 or more than 1 standard deviation, relative to practice in the art. When particular values are described in this application and claims, unless otherwise specified, the term "about" means within an acceptable range of error for the particular value.

[0018] "Administration" or "administering" refers to the process of providing (i.e., administering) a pharmaceutical composition to a subject, or alternatively, to a subject receiving the pharmaceutical composition. The pharmaceutical compositions disclosed herein can be administered locally by a variety of methods. For example, intramuscular, intradermal, subcutaneous, intrathecal, intraperitoneal, topical (transdermal), ophthalmic, and implantation (e.g., of a polymeric implant or sustained release device such as a mini-osmotic pump) can all be suitable routes of administration.

[0019] "Relief" means a reduction in the occurrence of headache pain or any of the symptoms or causes of a condition or disorder. Relief therefore includes some reduction, a significant reduction, near complete reduction, and complete reduction.

[0020] "Animal protein-free" means the absence of blood-derived, blood pool-derived, and other animal-derived products or compounds. "Animal" means mammals (such as humans), birds, reptiles, fish, insects, arachnids, or other animal species. "Animal" excludes microorganisms such as bacteria. Thus, an animal protein-free pharmaceutical composition can contain a botulinum neurotoxin. For example, an "animal protein-free" pharmaceutical composition means a pharmaceutical composition that is substantially free, essentially free, or completely free of other animal-derived proteins, such as serum-derived albumin, gelatin, and immunoglobulins. An example of an animal protein-free pharmaceutical composition is one that comprises or consists of a botulinum toxin (as the active ingredient) and a suitable polysaccharide as a stabilizer or excipient.

[0021] "Antioxidants" refers to any compound that protects an active ingredient from reacting with oxygen. Antioxidants can be broadly classified into three categories: (i) sacrificial antioxidants, such as ascorbic acid and sulfites, which can scavenge oxygen by reacting with it more readily than a specific active ingredient; (ii) chain terminators, such as methionine, NAC, glutathione, lipoic acid, butylated hydroxytoluene (BHT), and cysteine, which are molecules that form stable radicals due to a weak bond to a hydrogen atom that is attacked during radical chain propagation by consuming oxygen; and (iii) chelating agents, such as EDTA, EGTA, and DTPA, and their analogs, which reduce the catalytic activity of transition metals by forming complexes with the metals.

[0022] "Biological activity" describes the beneficial or harmful effects of a drug on an organism. When a drug is a complex chemical mixture, this activity is exerted by the active ingredient of the substance, but may be modified by other constituents. Biological activity can be measured by the in vivo LD, either as efficacy or as toxicity. 50 or ED 50 The activity of the compositions of the present invention can be measured using any suitable assay and is not limited to activity measured by CBPA. The potency described in the examples also includes potency as measured using LD50.

[0023] "Botulinum toxin" refers to neurotoxins produced by Clostridium botulinum, as well as botulinum toxins (or their heavy or light chains) recombinantly produced by non-clostridial species. As used herein, the term "botulinum toxin" includes botulinum toxin serotypes A, B, C, D, E, F, and G, and their subtypes and any of their subtypes, or, in each case, any genetically engineered protein, analog, derivative, homolog, part, subpart, mutant, or version of any of the foregoing. As used herein, "botulinum toxin" also includes "modified botulinum toxin." Furthermore, as used herein, "botulinum toxin" also includes botulinum toxin complexes (e.g., 300-, 600-, and 900-kDa complexes) and the neurotoxin component of botulinum toxin (150 kDa) without the complex protein attached.

[0024] "Clostridial toxin" refers to any toxin produced by a clostridial toxin strain that can enter a cell and carry out its general cellular machinery by binding the clostridial toxin to low- and high-affinity clostridial toxin receptors, internalization of the toxin / receptor complex, translocation of the clostridial toxin light chain into the cytoplasm, and enzymatic modification of the clostridial toxin substrate. Non-limiting examples of clostridial toxins include botulinum toxins such as BoNT / A, BoNT / B, BoNT / Ci, BoNT / D, BoNT / E, BoNT / F, BoNT / G, tetanus toxin (TeNT), Baratii toxin (BaNT), and Butyricum toxin (BuNT). BoNT / C2 cytotoxin and BoNT / C3 cytotoxin, which are not neurotoxins, are excluded from the term "clostridial toxin." The clostridial toxins disclosed herein include, but are not limited to, naturally occurring clostridial toxin variants, such as clostridial toxin isoforms and clostridial toxin subtypes, non-naturally occurring clostridial toxin variants, such as conservative clostridial toxin variants, non-conservative clostridial toxin variants, clostridial toxin chimeric variants, and active clostridial toxin fragments thereof, or any combination thereof. The clostridial toxins disclosed herein also include clostridial toxin complexes. As used herein, the term "clostridial toxin complex" refers to a complex comprising a clostridial toxin and a non-toxin-associated protein (NAP), such as a botulinum toxin complex, a tetanus toxin complex, a Baratii toxin complex, and a butyricum toxin complex. Non-limiting examples of clostridial toxin complexes include those produced by Clostridium botulinum, such as the 900 kDa BoNT / A complex, the 500 kDa BoNT / A complex, the 300 kDa BoNT / A complex, the 500 kDa BoNT / B complex, the 500 kDa BoNT / C1 complex, the 500 kDa BoNT / D complex, the 300 kDa BoNT / D complex, the 300 kDa BoNT / E complex, and the 300 kDa BoNT / F complex.

[0025] A "clostridial toxin active ingredient" refers to a molecule containing any portion of a clostridial toxin that exerts an effect upon or after administration to a subject or patient. As used herein, the term "clostridial toxin active ingredient" encompasses a clostridial toxin complex containing an approximately 150 kDa clostridial toxin and other proteins collectively referred to as non-toxin-associated proteins (NAPs), an approximately 150 kDa clostridial toxin alone, or a modified clostridial toxin (e.g., a retargeted clostridial toxin).

[0026] "Deformity" means any cosmetic, physical, or functional irregularity, defect, abnormality, imperfection, deformity, depression, or distortion.

[0027] "Effective amount," as applied to a biologically active ingredient, refers to the amount of the ingredient that is generally sufficient to bring about a desired change in a subject. For example, if the desired effect is a reduction in autoimmune disorder symptoms, an effective amount of the ingredient is an amount that causes at least a substantial reduction in the autoimmune disorder symptoms without causing significant toxicity.

[0028] When used in reference to the amount of an excipient, or a particular combination of excipients, added to a clostridial toxin composition, "effective amount" refers to the amount of each excipient necessary to achieve the desired initial recovery potential of the clostridial toxin active ingredient. In aspects of this embodiment, an effective amount of an excipient or combination of excipients results in, for example, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% initial recovery potential. In other aspects of this embodiment, a therapeutically effective concentration of a clostridial toxin active ingredient reduces symptoms associated with the treated diet by, for example, at most 10%, at most 20%, at most 30%, at most 40%, at most 50%, at most 60%, at most 70%, at most 80%, at most 90%, or at most 100%.

[0029] "Heavy chain" means the heavy chain of a botulinum neurotoxin, which has a molecular weight of about 100 kDa and is sometimes referred to as the H chain or H.

[0030] He refers to a fragment (approximately 50 kDa) derived from the heavy chain of a botulinum neurotoxin approximately equivalent to the carboxyl-terminal segment of the heavy chain, or a portion of the intact heavy chain corresponding to that fragment, which is immunogenic and is believed to contain the portion of native or wild-type botulinum neurotoxin responsible for high-affinity, presynaptic binding to motor neurons.

[0031] H N means a fragment (approximately 50 kDa) derived from the H chain of a botulinum neurotoxin that is approximately equivalent to the amino-terminal segment of the H chain, or a portion of the H chain corresponding to that complete fragment, which is believed to contain the portion of native or wild-type botulinum neurotoxin that is involved in the translocation of the L chain across the intracellular endosomal membrane.

[0032] "Light chain" refers to the light chain of a clostridial neurotoxin, which has a molecular weight of approximately 50 kDa and may also be referred to as the L chain, L, or the proteolytic domain (amino acid sequence) of a botulinum neurotoxin.

[0033] LH N or LH N is a fragment derived from a clostridial neurotoxin containing an L chain, or N By "heterologous clostridial neurotoxin" is meant a functional fragment thereof bound to a domain, which can be obtained from an intact clostridial neurotoxin by proteolysis to remove or modify the He domain.

[0034] "Implant" means a controlled-release (e.g., pulsed or continuous release) composition or drug delivery system. An implant can be, for example, injected, inserted, or implanted into the human body.

[0035] A "liquid composition," "liquid pharmaceutical composition," or "liquid formulation" refers to a pharmaceutically active formulation of a drug or biologically active ingredient that can be stored for an extended period of time in a liquid pharmaceutical excipient, such as a buffer, for immediate use by the clinician as needed. Liquid pharmaceutical compositions are manufactured without the lyophilization process.

[0036] "Topical administration" means the direct administration of a pharmaceutical agent to or within or near the surface of an animal's body where the biological effect of the pharmaceutical agent is desired, for example, by intramuscular administration, or intradermal or subcutaneous administration, or transdermal administration. Topical administration excludes systemic routes of administration, such as intravenous or oral administration. Transdermal administration is a type of topical administration in which a pharmaceutical agent is applied to the patient's skin.

[0037] "Lyophilization protectant" or "lyoprotectant" means a substance included in a lyophilized formulation to protect a clostridial toxin active ingredient during the lyophilization process. Lyophilization protectants include, for example, polyhydroxy compounds such as sugars (monosaccharides, disaccharides, and polysaccharides), polyalcohols, and derivatives thereof. Exemplary lyoprotectants that can be used with the lyophilized formulations disclosed herein include sucrose, trehalose, mannitol, sorbitol, glucose, raffinose, maltose, glycerol, lactose, fructose, galactose, and combinations thereof.

[0038] A "lyophilized composition," "lyophilized pharmaceutical composition," "lyophilized formulation," or "solid composition" refers to a formulation containing a clostridial toxin active ingredient that has been subjected to a lyophilization, freeze-drying, or vacuum-drying process and can be reconstituted with a reconstitution medium, such as saline or water, prior to administration to a patient. A lyophilized composition can be a freeze-dried composition or a vacuum-dried composition.

[0039] "Modified botulinum toxin" refers to a botulinum toxin in which at least one of its amino acids has been deleted, modified, or substituted compared to a native botulinum toxin. In addition, a modified botulinum toxin can be a recombinantly produced neurotoxin or a recombinantly produced derivative or fragment of a neurotoxin. A modified botulinum toxin retains at least one biological activity of a native botulinum toxin, such as the ability to bind to a botulinum toxin receptor or inhibit neurotransmitter release from neurons. One example of a modified botulinum toxin is a botulinum toxin having a light chain derived from one botulinum toxin serotype (e.g., type A) and a heavy chain derived from a different botulinum toxin serotype (e.g., type B). Another example of a modified botulinum toxin is a botulinum toxin conjugated to a neurotransmitter, such as substance P.

[0040] "Mutation" refers to a structural modification of a naturally occurring protein or nucleic acid sequence. For example, in the case of a nucleic acid mutation, the mutation can be a deletion, addition, or substitution of one or more nucleotides in the DNA sequence. In the case of a protein sequence mutation, the mutation can be a deletion, addition, or substitution of one or more amino acids in the protein sequence. For example, a particular amino acid comprising a protein sequence can be substituted with another amino acid, such as an amino acid selected from the group consisting of alanine, asparagine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, tyrosine, or any other natural or unnatural amino acid or chemically modified amino acid. Mutations to a protein sequence can be the result of a mutation to a DNA sequence that, when transcribed and the resulting mRNA translated, produces a mutant protein sequence. Mutations to a protein sequence can also be made by fusing a peptide sequence containing the desired mutation to the desired protein sequence.

[0041] "Patient" means a human or non-human subject receiving medical or veterinary care. Accordingly, the compositions disclosed herein can be used to treat any animal, such as a mammal.

[0042] "Administer peripherally" or "peripheral administration" means subcutaneous, intradermal, transdermal, or subcutaneous administration, but excludes intramuscular administration. "Peripheral" means within a subcutaneous location and excludes visceral locations.

[0043] "Pharmaceutical composition" means a composition comprising a Clostridial toxin active ingredient, e.g., a botulinum toxin, and at least one additional ingredient, e.g., a stabilizer or excipient. A pharmaceutical composition is thus a formulation suitable for diagnostic or therapeutic administration to a subject, such as a human patient. A pharmaceutical composition can be, for example, a lyophilized or vacuum-dried form, a solution formed after reconstitution of a lyophilized or vacuum-dried pharmaceutical composition, or a solution or solid that does not require reconstitution.

[0044] "Pharmacologically acceptable excipient" is synonymous with "pharmacological excipient" or "excipient" and refers to any excipient that has no substantial long-term or permanent adverse effects when administered to a mammal, and includes compounds such as stabilizers, bulking agents, cryoprotectants, lyoprotectants, additives, solvents, carriers, diluents, or adjuvants. Excipients are generally mixed with the active ingredient or are capable of diluting or encapsulating the active ingredient, and may be solid, semi-solid, or liquid. It is also envisioned that pharmaceutical compositions containing a clostridial toxin active ingredient can include one or more pharmaceutically acceptable excipients that facilitate processing of the active ingredient into a pharmaceutically acceptable composition. To the extent that any pharmacologically acceptable excipient is incompatible with the clostridial toxin active ingredient, its use in a pharmaceutically acceptable composition is contemplated. Non-limiting examples of pharmacologically acceptable excipients are described, for example, in Pharmaceutical Dosage Forms and Drug Delivery Systems (Howard C. Ansel et al., eds., Lippincott Williams & Wilkins Publishers, 7th ed.1999);Remington:The Science and Practice of Pharmacy(Alfonso R. Gennaro ed., Lippincott, Williams & Wilkins,20 th ed.2000);Goodman & Gilman's The Pharmacological Basis of Therapeutics(Joel G. Hardman et al, eds., McGraw-Hill Professional,10 th ed.2001); and Handbook of Pharmaceutical Excipients (Raymond C. Rowe et al, APhA Publications, 4 th edition 2003), each of which is incorporated herein by reference in its entirety.

[0045] The components of the pharmaceutical composition may be included as a single composition (i.e., all components except any necessary reconstitution fluid are present at the time of initial formulation of the pharmaceutical composition) or as a two-component system, e.g., a vacuum-dried composition, reconstituted with a reconstitution medium that may contain components not present in the initial formulation of the pharmaceutical composition. Two-component systems can offer several benefits, such as allowing the mixing of components that are not sufficiently compatible with the first component of the two-component system for long-term storage. For example, the reconstitution medium may contain a preservative that provides sufficient protection against microbial growth during the period of use, e.g., a one-week refrigerated storage period, but is not present during a two-year frozen storage period during which the toxin may be degraded. Other components that are not long-term compatible with the botulinum toxin or other components can be mixed in this manner, i.e., added to a second medium (e.g., the reconstitution medium) near the time of use. Pharmaceutical compositions can also contain preservatives such as benzyl alcohol, benzoic acid, phenol, parabens, and sorbic acid. Pharmaceutical compositions can include excipients such as, for example, surfactants, dispersing agents, inert diluents, granulating and disintegrating agents, binders, lubricants, preservatives, physiologically degradable compositions such as gelatin, aqueous vehicles and solvents, oily vehicles and solvents, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, buffers, salts, thickening agents, fillers, antioxidants, stabilizers, and pharmaceutically acceptable polymeric or hydrophobic materials, as well as other ingredients known in the art and described, for example, in Genaro, ed., 1985, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., which is incorporated herein by reference.

[0046] "Polysaccharide" means a polymer of three or more sugar molecule monomers, which may be the same or different.

[0047] "Stabilizer," "stabilizer," or "stabilizing agent" means a substance that acts to stabilize a clostridial toxin active ingredient such that the efficacy of the pharmaceutical composition is increased relative to a destabilized composition.

[0048] "Stabilizers" can include excipients, and can include protein and non-protein molecules.

[0049] "Surfactant" refers to a natural or synthetic amphiphilic compound. Surfactants can be nonionic, zwitterionic, or ionic. Non-limiting examples of surfactants include poloxamers, polysorbates, and combinations thereof.

[0050] "Therapeutic formulation" means a formulation that can be used to treat and thereby alleviate a disorder or disease, such as, for example, an injury or disease characterized by peripheral muscle hyperactivity (ie, spasticity).

[0051] As used herein, "TEM" is synonymous with "targeted exocytosis regulator" or "retargeting endopeptidase." Generally, TEMs comprise an enzymatic domain derived from a clostridial toxin light chain, a translocation domain derived from a clostridial toxin heavy chain, and a targeting domain. The targeting domain of a TEM provides an altered cellular targeting ability, targeting the molecule to a receptor other than the native clostridial toxin receptor utilized by the native clostridial toxin. This retargeting ability is achieved by replacing the naturally occurring binding domain of a clostridial toxin with a targeting domain that has binding activity to a non-clostridial toxin receptor. Although it binds to a non-clostridial toxin receptor, TEMs undergo all of the other steps in the intoxication process, including internalization of the TEM / receptor complex into the cytoplasm, formation of pores in the vesicle membrane and the double-stranded molecule, translocation of the enzymatic domain into the cytoplasm, and exerting a proteolytic effect on components of the SNARE complex of the target cell.

[0052] "Tonicity agent" or "tonicity agent" refers to a low molecular weight excipient included in a lyophilized or liquid formulation that imparts isotonicity. For example, disaccharides such as trehalose or sucrose, polyalcohols such as sorbitol or mannitol, monosaccharides such as glucose, and salts such as sodium chloride can function as isotonicity agents.

[0053] "Transdermal administration" excludes systemic administration of the neurotoxin. In other words, unlike conventional therapeutic transdermal methods, transdermal administration of a botulinum toxin results in a significant amount, e.g., a majority, of the neurotoxin not entering the patient's circulatory system.

[0054] "Treatment" means, for example, alleviating (or eliminating) at least one symptom of a condition or disorder, such as wrinkles, spasticity, depression, pain (e.g., headaches), bladder overactivity, etc., either temporarily or permanently.

[0055] As used herein, the term "unit" or "U" refers to an LD 50 LD refers to the dose determined by the cell-based potency assay (CBPA). 50 A dose is defined as the amount of a clostridial toxin active ingredient, clostridial toxin complex, or modified clostridial toxin that resulted in the death of 50% of mice injected with the clostridial toxin, clostridial toxin complex, or modified clostridial toxin. CBPA doses were determined as described in U.S. Patent No. 8,618,261, the assay details of which are incorporated herein by reference.

[0056] "Mutant" means a clostridial neurotoxin, such as wild-type botulinum toxin serotype A, B, C, D, E, F, or G, that has been modified by substitution, alteration, addition, or deletion of at least one amino acid relative to the wild-type botulinum toxin, and which is recognized by and internalized by a target cell and catalytically cleaves SNARE (SNAP (soluble NSF attachment protein) receptor) proteins within the target cell.

[0057] An example of a variant neurotoxin component can include a variant light chain of a botulinum toxin having one or more amino acids substituted, modified, deleted, and / or added. The variant light chain may have the same or better ability to prevent exocytosis, e.g., release of neurotransmitter secretory vesicles. The biological effect of the variant may also be reduced compared to the parent chemical. For example, a variant light chain of a botulinum toxin type A having a deleted amino acid sequence may have a shorter biological persistence than the parent (i.e., native) botulinum toxin type A light chain.

[0058] In one embodiment, the composition does not contain a sugar or polyalcohol. Sugar is defined below. As used herein, the term "polyalcohol" is synonymous with "sugar alcohol," "polyhydric alcohol," and "polyol" and refers to a sugar derivative having an alcohol group (CHOH) instead of an aldehyde group (CHO), such as mannitol (derived from mannose), xylitol (derived from xylose), and lactitol (derived from lactulose). Non-limiting examples of polyols include glycol, glycerol, arabitol, erythritol, xylitol, maltitol, sorbitol (glucitol), mannitol, inositol, lactitol, galactitol (iditol), and isomalt. Other non-limiting examples of sugar excipients are described, for example, in Ansel (1999), Gennaro (2000), Hardman (2001), and Rowe (2003), each of which is incorporated herein by reference in its entirety. Pharmaceutical Composition

[0059] Certain embodiments of the present invention include pharmaceutical compositions comprising (or consisting of, or consisting essentially of) a Clostridial toxin active ingredient, such as a botulinum toxin, a disaccharide, a surfactant, and an antioxidant.

[0060] Aspects of the pharmaceutical compositions of the present invention provide, in part, a clostridial toxin active ingredient. As used herein, the term "clostridial toxin active ingredient" refers to a therapeutically effective concentration of a clostridial toxin active ingredient, such as a clostridial toxin complex, a clostridial toxin, a modified clostridial toxin, or a retargeted clostridial toxin. Botulinum toxin is a particularly preferred clostridial toxin active ingredient for use in the present invention. As used herein, the term "therapeutically effective concentration" is synonymous with "therapeutically effective amount," "effective amount," "effective dose," and "therapeutically effective dose," and refers to the minimum dose of a clostridial toxin active ingredient, most preferably a botulinum toxin, required to achieve the desired therapeutic effect, including a dose sufficient to reduce symptoms associated with the diet being treated. In aspects of this embodiment, a therapeutically effective concentration of a clostridial toxin active ingredient, most preferably a botulinum toxin, reduces symptoms associated with the treated meal, e.g., by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%. In other aspects of this embodiment, a therapeutically effective concentration of a clostridial toxin active ingredient, most preferably a botulinum toxin, reduces symptoms associated with the treated meal, e.g., by up to 10%, up to 20%, up to 30%, up to 40%, up to 50%, up to 60%, up to 70%, up to 80%, up to 90%, or up to 100%.

[0061] It is contemplated that any amount of clostridial toxin active ingredient, most preferably botulinum toxin, can be added in formulating the clostridial toxin pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of the clostridial toxin active ingredient is recoverable. In aspects of this embodiment, the amount of clostridial toxin active ingredient, most preferably botulinum toxin, added to the formulation is at least 0.1 U / mL, at least 1.0 U / mL, at least 10 U / mL, at least 50 U / mL, at least 100 U / mL, at least 200 U / mL, or at least 1000 U / mL. In another aspect of this embodiment, the amount of clostridial toxin active ingredient, most preferably botulinum toxin, added to the formulation is at most 0.1 U / mL, at most 1.0 U / mL, at most 10 U / mL, at most 50 U / mL, at most 100 U / mL, at most 200 U / mL, or at most 1000 U / mL. In yet another aspect of this embodiment, the amount of clostridial toxin active ingredient, most preferably botulinum toxin, added to the formulation is from about 0.1 U / mL to about 1000 U / mL, or from about 1.0 U / mL to about 1000 U / mL. In yet another aspect of this embodiment, the amount of clostridial toxin active ingredient, most preferably botulinum toxin, added to the formulation is from about 0.001 U / mL to about 100 U / mL, from about 0.01 U / mL to about 100 U / mL, from about 0.1 U / mL to about 100 U / mL, or from about 1.0 U / mL to about 100 U / mL.

[0062] In another aspect of this embodiment, the amount of clostridial toxin active ingredient, most preferably botulinum toxin, added to the formulation is at least 1.0 pg, at least 10 pg, at least 100 pg, at least 1.0 ng, at least 10 ng, at least 100 ng, at least 1.0 μg, at least 10 μg, at least 100 μg, or at least 1.0 mg. In yet another aspect of this embodiment, the amount of clostridial toxin active ingredient, most preferably botulinum toxin, added to the formulation is at most 1.0 pg, at most 10 pg, at most 100 pg, at most 1.0 ng, at most 10 ng, at most 100 ng, at most 1.0 μg, at most 10 μg, at most 100 μg, or at most 1.0 mg. In yet another aspect of this embodiment, the amount of a clostridial toxin active ingredient, most preferably a botulinum toxin, added to the formulation is about 1.0 pg to about 10 μg, about 10 pg to about 10 μg, about 100 pg to about 10 μg, about 1.0 ng to about 10 μg, about 10 ng to about 10 μg, or about 100 ng to about 10 μg. In yet another aspect of this embodiment, the amount of a clostridial toxin active ingredient, most preferably a botulinum toxin, added to the formulation is about 1.0 pg to about 1.0 μg, about 10 pg to about 1.0 μg, about 100 pg to about 1.0 μg, about 1.0 ng to about 1.0 μg, about 10 ng to about 1.0 μg, or about 100 ng to about 1.0 μg. In a further aspect of this embodiment, the amount of a clostridial toxin active ingredient, most preferably a botulinum toxin, added to the formulation is about 1.0 pg to about 5.0 μg, about 10 pg to about 5.0 μg, about 100 pg to about 5.0 μg, about 1.0 ng to about 5.0 μg, about 10 ng to about 5.0 μg, or about 100 ng to about 5.0 μg. In a further aspect of this embodiment, the amount of a clostridial toxin active ingredient, most preferably a botulinum toxin, added to the formulation is about 1.0 pg to about 10 μg, about 10 pg to about 10 μg, about 100 pg to about 10 μg, about 1.0 ng to about 10 μg, about 10 ng to about 10 μg, or about 100 ng to about 10 pg.

[0063] In aspects of this embodiment, the clostridial toxin pharmaceutical composition comprises BoNT / A, BoNT / B, BoNT / Ci, BoNT / D, BoNT / E, BoNT / F, BoNT / G, a BoNT mosaic (e.g., BoNT / DC, BoNT / CD, and BoNT / FA), TeNT, BaNT, or BuNT. In another embodiment, the clostridial toxin pharmaceutical composition comprises a clostridial toxin mutant as the clostridial toxin active ingredient. In aspects of this embodiment, the clostridial toxin pharmaceutical composition comprises a naturally occurring clostridial toxin mutant or a non-naturally occurring clostridial toxin mutant. In another aspect of this embodiment, the clostridial toxin pharmaceutical composition comprises a BoNT / A mutant, a BoNT / B mutant, a BoNT / Ci mutant, a BoNT / D mutant, a BoNT / E mutant, a BoNT / F mutant, a BoNT / G mutant, a TeNT mutant, a BaNT mutant, or a BuNT mutant, wherein the mutant is either a naturally occurring mutant or a non-naturally occurring mutant.

[0064] Some aspects of the pharmaceutical compositions of the present invention provide a clostridial toxin complex as a clostridial toxin active ingredient. As used herein, the term "clostridial toxin complex" refers to a complex containing a clostridial toxin and related NAP, such as a botulinum toxin complex, a tetanus toxin complex, a Baratii toxin complex, and a butyricum toxin complex. Non-limiting examples of clostridial toxin complexes include those produced by Clostridium botulinum, such as the 900 kDa BoNT / A complex, the 500 kDa BoNT / A complex, the 300 kDa BoNT / A complex, the 500 kDa BoNT / B complex, the 500 kDa BoNT / C1 complex, the 500 kDa BoNT / D complex, the 300 kDa BoNT / D complex, the 300 kDa BoNT / E complex, and the 300 kDa BoNT / F complex. Clostridial toxin complexes can be purified using the methods described in Schantz (1992), supra, and Hui Xiang et al., Animal Product Free System and Process for Purifying a Botulinum Toxin, U.S. Patent No. 7,354,740, each of which is incorporated herein by reference in its entirety. Clostridial toxin complexes are available, for example, from List Biological Laboratories, Inc. (Campbell, California), the Centre for Applied Microbiology and Research (Porton Down, UK), Wako (Osaka, Japan), and Sigma Chemicals (St Louis, Missouri).

[0065] Some aspects of the pharmaceutical compositions of the present invention provide a non-protein excipient. As used herein, the term "non-protein excipient" refers to any excipient that is not a polypeptide containing at least 15 amino acids. Any non-protein excipient is useful in formulating the clostridial toxin pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of the clostridial toxin active ingredient, most preferably a botulinum toxin, is recovered using the non-protein excipient.

[0066] Aspects of the pharmaceutical compositions of the present invention provide, in part, sugars. As used herein, the term "sugar" refers to compounds containing 1 to 10 monosaccharide units, e.g., monosaccharides, disaccharides, trisaccharides, and oligosaccharides containing 4 to 10 monosaccharide units. Any sugar is useful in formulating the clostridial toxin pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of a clostridial toxin active ingredient, most preferably a botulinum toxin, is recovered using the sugar. In some embodiments, for example, in a lyophilized composition, a sugar can function as a lyoprotectant. In some other embodiments, for example, in a lyophilized or liquid formulation, a sugar can function as a tonicity agent. Monosaccharides include polyhydroxyaldehydes or polyhydroxyketones having three or more carbon atoms, such as aldoses, dialdoses, aldoketoses, ketoses, and diketoses, as well as cyclic forms, deoxysugars, and aminosugars, and derivatives thereof, when the parent monosaccharide contains a (potential) carbonyl group. Monosaccharides include trioses such as glyceraldehyde and dihydroxyacetone; tetroses such as erythrose, erythrulose, and threose; pentoses such as arabinose, lyxose, ribose, ribulose, xylose, and xylulose; hexoses such as allose, altrose, fructose, fucose, galactose, glucose, gulose, idose, mannose, psicose, rhamnose, sorbose, tagatose, talose, and trehalose; heptoses such as sedoheptulose and mannoheptulose; octose such as octulose and 2-keto-3-deoxy-mannooctonate; nonoses such as sialose; and decose. Oligosaccharides are compounds in which at least two types of monosaccharide units are linked by glycosidic bonds. Depending on the number of units, they are called disaccharides, trisaccharides, tetrasaccharides, pentasaccharides, hexasaccharides, heptasaccharides, octasaccharides, nonasaccharides, decasaccharides, etc. Oligosaccharides can be unbranched, branched, or cyclic. Common disaccharides include, but are not limited to, sucrose, lactose, maltose, trehalose, cellobiose, gentiobiose, kojibiose, laminaribiose, mannobiose, melibiose, nigerose, rutinose, and xylobiose.Common trisaccharides include, but are not limited to, raffinose, acarbose, maltotriose, and melezitose. Other non-limiting examples of sugar excipients for specific applications are described, for example, in Ansel (1999), supra; Gennaro (2000), supra; Hardman (2001), supra; and Rowe (2003), supra, each of which is incorporated herein by reference in its entirety.

[0067] In some embodiments, a clostridial toxin pharmaceutical composition comprises a sugar. In aspects of this embodiment, the clostridial toxin pharmaceutical composition comprises a monosaccharide. In other aspects of this embodiment, the clostridial toxin pharmaceutical composition comprises a disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, hexasaccharide, heptasaccharide, octasaccharide, nonasaccharide, or decasaccharide. In yet other aspects of this embodiment, the clostridial toxin pharmaceutical composition comprises an oligosaccharide comprising 2 to 10 monosaccharide units. Trehalose and sucrose are particularly preferred sugars for use in the present invention.

[0068] Any amount of sugar, preferably trehalose or sucrose, is useful in formulating the clostridial toxin pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of the clostridial toxin active ingredient, most preferably a botulinum toxin, is recovered using the amount of sugar. In aspects of this embodiment, the amount of sugar, preferably trehalose or sucrose, added to the formulation is about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0% (w / w), or , about 5.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), about 10% (w / w), about 15% (w / w), about 20% (w / w), about 25% (w / w), about 30% (w / w), or about 35% (w / w). In other aspects of this embodiment, the amount of sugar, preferably trehalose or sucrose, added to the formulation is at least 0.1% (w / w), at least 0.5% (w / w), at least 1.0% (w / w), at least 1.5% (w / w), at least 2.0% (w / w), at least 2.5% (w / w), at least 3.0% (w / w), at least 3.5% (w / w), at least 4.0% (w / w), at least 4.5% (w / w), at least 5.0 ... 0.5% (w / w), at least 6.0% (w / w), at least 6.5% (w / w), at least 7.0% (w / w), at least 7.5% (w / w), at least 8.0% (w / w), at least 8.5% (w / w), at least 9.0% (w / w), at least 9.5% (w / w), at least 10% (w / w), at least 15% (w / w), at least 20% (w / w), at least 25% (w / w), at least 30% (w / w), or at least 35% (w / w).In other aspects of this embodiment, the amount of sugar, preferably trehalose or sucrose, added to the formulation is at most 0.1% (w / w), at most 0.5% (w / w), at most 1.0% (w / w), at most 1.5% (w / w), at most 2.0% (w / w), at most 2.5% (w / w), at most 3.0% (w / w), at most 3.5% (w / w), at most 4.0% (w / w), at most 4.5% (w / w), at most 5.0% (w / w), at most 6.0% (w / w), at most 7.0% (w / w), at most 8.0% (w / w), at most 9.0% (w / w), at most 10.0% (w / w), at most 11.0% (w / w), at most 12.0% (w / w), at most 13.0% (w / w), at most 14.0% (w / w), at most 15.0% (w / w), at most 16.0% (w / w), at most 17.0% (w / w), at most 18.0% (w / w), at most 19.0% (w / w), at most 20.0% (w / w), at most 21.0% (w / w), at most 22.0% (w / w), at most 23.0% (w / w), at most 24.0% (w / w), at most 25.0% (w / w), at most 26.0% (w / w), at most 27.0% (w / w), at most 28.0% (w / w), at most 29.0% (w / w), at most 30.0% (w / w), at most 31.0% (w / w), at most 32.0% (w / w), at most 3 at most 5.5% (w / w), at most 6.0% (w / w), at most 6.5% (w / w), at most 7.0% (w / w), at most 7.5% (w / w), at most 8.0% (w / w), at most 8.5% (w / w), at most 9.0% (w / w), at most 9.5% (w / w), at most 10% (w / w), at most 15% (w / w), at most 20% (w / w), at most 25% (w / w), at most 30% (w / w), or at most 35% (w / w).

[0069] In some embodiments, a clostridial toxin pharmaceutical composition of the invention comprises a disaccharide, preferably trehalose or sucrose. Common disaccharides include, but are not limited to, sucrose, lactose, maltose, trehalose, cellobiose, gentiobiose, kojibiose, laminaribiose, mannobiose, melibiose, nigerose, rutinose, and xylobiose. In an aspect of this embodiment, the clostridial toxin pharmaceutical composition comprises sucrose. In one particular embodiment, the clostridial toxin pharmaceutical composition comprises trehalose. In aspects of this embodiment, the amount of disaccharide, preferably trehalose or sucrose, added to the formulation is about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0% (w / w), about 6.0% (w / w), about 7.0% (w / w), about 8.0% (w / w), about 9.0% (w / w), about 10.0% (w / w), about 11.0% (w / w), about 12.0% (w / w), about 13.0% (w / w), about 14.0% (w / w), about 15.0% (w / w), about 16.0% (w / w), about 17.0% (w / w), about 18.0% (w / w), about 19.0% (w / w), about 20.0% (w / w), about 21.0% (w / w), about 22.0% (w / w), about 23.0% (w / w), about 24.0% (w / w), about 25.0% (w / w), about 26.0% (w / w), about 27.0% (w / w), about 28.0% (w / w), about 29.0% (w / w), about 30.0% (w / w), about 31.0% (w / w), about 32.0% (w / w), about 33.0% (w / w), about 34.0% (w / w), about 35.0% (w / w), about 36.0 (w / w), about 5.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), about 10% (w / w), about 15% (w / w), about 20% (w / w), about 25% (w / w), about 30% (w / w), or about 35% (w / w).

[0070] Aspects of the pharmaceutical compositions of the present invention provide, in part, a surfactant. As used herein, the term "surfactant" refers to a natural or synthetic amphiphilic compound. Surfactants can be nonionic, zwitterionic, or ionic. Any surfactant is useful in formulating the clostridial toxin pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of the clostridial toxin active ingredient, most preferably a botulinum toxin, is recovered using the amount of surfactant. Non-limiting examples of surfactants include polysorbates such as polysorbate 20 (TWEEN® 20), polysorbate 40 (TWEEN® 40), polysorbate 60 (TWEEN® 60), polysorbate 61 (TWEEN® 61), polysorbate 65 (TWEEN® 65), polysorbate 80 (TWEEN® 80), and polysorbate 81 (TWEEN® 81); poloxamer 124 (PLURONIC® L44), poloxamer 181 (PLURONIC® L61), poloxamer 182 (PLURONIC® L62), poloxamer 184 (PLURONIC® L64); Poloxamers (polyethylene-polypropylene copolymers) such as Poloxamer 188 (PLURONIC® F68), Poloxamer 237 (PLURONIC® F87), Poloxamer 338 (PLURONIC® L108), Poloxamer 407 (PLURONIC® F127), polyoxyethylene glycol dodecyl ethers such as BRIJ® 30 and BRIJ® 35; 2-dodecoxyethanol (LUBROL®-PX); polyoxyethylene octylphenyl ether (TRITON® X-100); sodium dodecyl sulfate (SDS); solutol HS15; 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate (CHAPS); 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-l-propanesulfonate (CHAPSO); sucrose monolaurate; and sodium cholate.Other non-limiting examples of surfactant excipients are described, for example, in Ansel (1999), supra; Gennaro (2000), supra; Hardman (2001), supra; and Rowe (2003), supra, each of which is incorporated herein by reference in its entirety. Poloxamer 188 is a particularly preferred surfactant in the present invention.

[0071] Thus, in one embodiment, a clostridial toxin pharmaceutical composition comprises a surfactant, preferably poloxamer 188. In aspects of this embodiment, the clostridial toxin pharmaceutical composition comprises a polysorbate, poloxamer, polyoxyethylene glycol dodecyl ether, 2-dodecoxyethanol, polyoxyethylene octylphenyl ether, sodium dodecyl sulfate, 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate, 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-l-propanesulfonate, sucrose monolaurate, or sodium cholate.

[0072] Any amount of surfactant, preferably poloxamer 188, is useful in formulating the clostridial toxin pharmaceutical compositions disclosed herein, provided that a therapeutically effective amount of the clostridial toxin active ingredient, preferably a botulinum toxin, is recovered using that amount of surfactant. In aspects of this embodiment, the amount of surfactant, preferably poloxamer 188, added to the formulation is about 0.01% (w / w), about 0.02% (w / w), about 0.03% (w / w), about 0.04% (w / w), about 0.05% (w / w), about 0.06% (w / w), about 0.07% (w / w), about 0.08% (w / w), about 0.09% (w / w), about 0.1% (w / w), about 0.5% (w / w), about 1.0% (w / w), about 1.5% (w / w), about 2.0% (w / w), about 2.5% (w / w), about 3.0% (w / w), about 3.5% (w / w), about 4.0% (w / w), about 4.5% (w / w), about 5.0% (w / w), about 5.5% (w / w), about 6.0% (w / w), about 6.5% (w / w), about 7.0% (w / w), about 7.5% (w / w), about 8.0% (w / w), about 8.5% (w / w), about 9.0% (w / w), about 9.5% (w / w), about 10% (w / w), about 15% (w / w), about 20% (w / w), about 25% (w / w), about 30% (w / w), or about 35% (w / w).In other aspects of this embodiment, the amount of surfactant, preferably poloxamer 188, added to the formulation is at least 0.01% (w / w), at least 0.02% (w / w), at least 0.03% (w / w), at least 0.04% (w / w), at least 0.05% (w / w), at least 0.06% (w / w), at least 0.07% (w / w), at least 0.08% (w / w), at least 0.09% (w / w), at least 0.1% (w / w), at least 0.5% (w / w), at least 1.0% (w / w), at least 1.5% (w / w), at least 2.0% (w / w), at least 2.5% (w / w), at least 3.0% (w / w), at least 4.0% (w / w), at least 5.0% (w / w), at least 6.0% (w / w), at least 7.0% (w / w), at least 8.0% (w / w), at least 9.0% (w / w), at least 10.0% (w / w), at least 11.0% (w / w), at least 12.0% (w / w), at least 13.0% (w / w), at least 14.0% (w / w), at least 15.0% (w / w), at least 16.0% (w / w), at least 17.0% (w / w), at least 18.0% (w / w), at least 19.0% (w / w), at least 20.0% (w / w), at least 21.0% (w / w), at least 22.0% (w / w), at least 23.0% (w / w), at least 24.0% (w / w), at least 25.0% (w / w), at least (w / w), at least 3.5% (w / w), at least 4.0% (w / w), at least 4.5% (w / w), at least 5.0% (w / w), at least 5.5% (w / w), at least 6.0% (w / w), at least 6.5% (w / w), at least 7.0% (w / w), at least 7.5% (w / w), at least 8.0% (w / w), at least 8.5% (w / w), at least 9.0% (w / w), at least 9.5% (w / w), at least 10% (w / w), at least 15% (w / w), at least 20% (w / w), at least 25% (w / w), at least 30% (w / w), or at least 35% (w / w).In yet another aspect of this embodiment, the amount of surfactant, preferably poloxamer 188, added to the formulation is at most 0.01% (w / w), at most 0.02% (w / w), at most 0.03% (w / w), at most 0.04% (w / w), at most 0.05% (w / w), at most 0.06% (w / w), at most 0.07% (w / w), at most 0.08% (w / w), at most 0.09% (w / w), at most 0.1% (w / w), at most 0.5% (w / w), at most 1.0% (w / w), at most 1.5% (w / w), at most 2.0% (w / w), at most 2.5% (w / w), at most 3.0% (w / w), at most 4.0% (w / w), at most 5.0% (w / w), at most 6.0% (w / w), at most 7.0% (w / w), at most 8.0% (w / w), at most 9.0% (w / w), at most 10.0% (w / w), at most 11.0% (w / w), at most 12.0% (w / w), at most 13.0% (w / w), at most 14.0% (w / w), at most 15.0% (w / w), at most 16.0% (w / w), at most 17.0% (w / w), at most 18.0% (w / w), at most 19.0% (w / w), at most 20.0% (w / w), at most 21.0% (w / w), at most 22.0% (w / w), at most 23.0% (w / w), at most 24.0% (w / w), at most 25.0% (w / w), at 0.0% (w / w), at most 3.5% (w / w), at most 4.0% (w / w), at most 4.5% (w / w), at most 5.0% (w / w), at most 5.5% (w / w), at most 6.0% (w / w), at most 6.5% (w / w), at most 7.0% (w / w), at most 7.5% (w / w), at most 8.0% (w / w), at most 8.5% (w / w), at most 9.0% (w / w), at most 9.5% (w / w), at most 10% (w / w), at most 15% (w / w), at most 20% (w / w), at most 25% (w / w), at most 30% (w / w), or at most 35% (w / w).

[0073] In some embodiments, a clostridial toxin pharmaceutical composition comprises a poloxamer, preferably poloxamer 188. Poloxamers that can be used with the pharmaceutical compositions of the invention include poloxamer 124 (PLURONIC® L44), poloxamer 181 (PLURONIC® L61), poloxamer 182 (PLURONIC® L62), poloxamer 184 (PLURONIC® L64), poloxamer 188 (PLURONIC® F68), poloxamer 237 (PLURONIC® F87), poloxamer 338 (PLURONIC® L108), and poloxamer 407 (PLURONIC® F127). In some embodiments, poloxamer 188 may be more advantageous.

[0074] In some embodiments, a clostridial toxin pharmaceutical composition comprises a polysorbate. Polysorbates that can be used with pharmaceutical compositions of the invention include polysorbate 20 (TWEEN® 20), polysorbate 40 (TWEEN® 40), polysorbate 60 (TWEEN® 60), polysorbate 61 (TWEEN® 61), polysorbate 65 (TWEEN® 65), polysorbate 80 (TWEEN® 80), and polysorbate 81 (TWEEN® 81). In some embodiments, polysorbate 20 may be advantageous over some other polysorbates.

[0075] Some aspects of the pharmaceutical compositions of the present invention provide at least one antioxidant. Non-limiting examples of antioxidants include, but are not limited to, methionine, cysteine, N-acetylcysteine (NAC), sodium disulfite, sodium thiosulfate, butylated hydroxyanisole, butylated hydroxytoluene, vitamin E, and analogs such as Trolox C; chelating agents such as EDTA (ethylenediaminetetraacetic acid sodium salt), EGTA (ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid), DTPA (diethylenetriaminepentaacetic acid), analogs or derivatives thereof, and combinations thereof. Methionine and NAC are particularly preferred antioxidants for use in the present invention. NAC is particularly preferably used together with EDTA in certain formulations of the present invention. In certain other lyophilized formulations of the present invention, NAC is the only antioxidant used. In still other certain formulations of the present invention, methionine is the only antioxidant used. In aspects of these embodiments, the amount of antioxidant, preferably methionine or NAC, added to the formulation is in the range of about 0.01% (w / w) to about 0.10% (w / w). When used with NAC, EDTA is preferably added in an amount of about 0.01% (w / w) to about 0.10% (w / w).

[0076] It is further contemplated that the clostridial toxin pharmaceutical compositions disclosed herein can optionally contain other pharmaceutically acceptable ingredients (or pharmaceutical components), including, but not limited to, buffers, preservatives, tonicity adjusting agents, salts, antioxidants, osmolality adjusting agents, emulsifiers, sweeteners, or flavoring agents. A variety of buffers and means for adjusting pH can be used to prepare the pharmaceutical compositions disclosed herein, provided that the resulting preparation is pharmaceutically acceptable. Such buffers include, but are not limited to, acetate buffer, borate buffer, citrate buffer, phosphate buffer, neutral buffered saline, and phosphate buffered saline. Histidine is a particularly preferred buffer. It is understood that an acid or base can be used to adjust the pH of a pharmaceutical composition as needed. Any buffered pH level can be useful in formulating a clostridial toxin pharmaceutical composition, provided that a therapeutically effective amount of the clostridial toxin active ingredient, preferably a botulinum toxin, is recovered using this effective pH level. In some aspects of this embodiment, the effective pH level is at least about pH 5.0, at least about pH 5.5, at least about pH 6.0, at least about pH 6.5, at least about pH 7.0, or about pH 7.5. In other aspects of this embodiment, the effective pH level is at most about pH 5.0, at most about pH 5.5, at most about pH 6.0, at most about pH 6.5, at most about pH 7.0, or at most about pH 7.5. In yet another aspect of this embodiment, the effective pH level is about pH 5.0 to about pH 8.0, the effective pH level is about pH 5.0 to about pH 7.0, the effective pH level is about pH 5.0 to about pH 6.0, the effective pH level is about pH 5.5 to about pH 8.0, the effective pH level is about pH 5.5 to about pH 7.0, the effective pH level is about pH 5.5 to about pH 5.0, the effective pH level is about pH 5.5 to about pH 7.5, and the effective pH level is about pH 5.5 to about pH 6.5. An effective pH of 5.5 to 6.0 is particularly preferred.

[0077] The pharmaceutical compositions disclosed herein can have a pH of about 5-8 upon reconstitution or injection. In certain embodiments, the compositions will have a pH of less than 8, such as 7.9, or 7.8, or 7.7, or 7.6, or 7.5, or 7.4, or 7.3, or 7.2, or 7.1, or 7.0, or 6.9, or 6.8, or 6.7, or 6.6, or 6.5, or 6.4, or 6.3, or 6.2, or 6.1, or 6.0, or 5.9, or 5.8, or 5.7, or 5.6, or 5.5, or 5.4, or 5.3, or 5.2, or 5.1. In some embodiments, the pH is in the range of 5-7. A pH of 5.5-6.5 is preferred, with a pH of 5.5-6.0 being particularly preferred.

[0078] Any concentration of buffering agent may be useful in formulating a clostridial toxin pharmaceutical composition, provided that a therapeutically effective amount of a clostridial toxin active ingredient, most preferably a botulinum toxin, is recovered using this effective concentration of buffering agent, preferably histidine. In aspects of this embodiment, the effective concentration of buffering agent, preferably histidine, is at least 0.1 mM, at least 0.2 mM, at least 0.3 mM, at least 0.4 mM, at least 0.5 mM, at least 0.6 mM, at least 0.7 mM, at least 0.8 mM, or at least 0.9 mM. In other aspects of this embodiment, the effective concentration of buffering agent, preferably histidine, is at least 1.0 mM, at least 2.0 mM, at least 3.0 mM, at least 4.0 mM, at least 5.0 mM, at least 6.0 mM, at least 7.0 mM, at least 8.0 mM, or at least 9.0 mM. In yet other aspects of this embodiment, the effective concentration of the buffering agent, preferably histidine, is at least 10 mM, at least 20 mM, at least 30 mM, at least 40 mM, at least 50 mM, at least 60 mM, at least 70 mM, at least 80 mM, or at least 90 mM. In yet other aspects of this embodiment, the effective concentration of the buffering agent, preferably histidine, is at least 100 mM, at least 200 mM, at least 300 mM, at least 400 mM, at least 500 mM, at least 600 mM, at least 700 mM, at least 800 mM, or at least 900 mM. In further aspects of this embodiment, the effective concentration of the buffering agent, preferably histidine, is at most 0.1 mM, at most 0.2 mM, at most 0.3 mM, at most 0.4 mM, at most 0.5 mM, at most 0.6 mM, at most 0.7 mM, at most 0.8 mM, or at most 0.9 mM. In yet other aspects of this embodiment, the effective concentration of the buffering agent, preferably histidine, is at most 1.0 mM, at most 2.0 mM, at most 3.0 mM, at most 4.0 mM, at most 5.0 mM, at most 6.0 mM, at most 7.0 mM, at most 8.0 mM, or at most 9.0 mM.In yet other aspects of this embodiment, the effective concentration of the buffering agent, preferably histidine, is at most 10 mM, at most 20 mM, at most 30 mM, at most 40 mM, at most 50 mM, at most 60 mM, at most 70 mM, at most 80 mM, or at most 90 mM. In yet other aspects of this embodiment, the effective concentration of the buffering agent, preferably histidine, is at most 100 mM, at most 200 mM, at most 300 mM, at most 400 mM, at most 500 mM, at most 600 mM, at most 700 mM, at most 800 mM, or at most 900 mM. In still further aspects of this embodiment, the effective concentration of the buffer, preferably histidine, is about 0.1 mM to about 900 mM, 0.1 mM to about 500 mM, 0.1 mM to about 100 mM, 0.1 mM to about 90 mM, 0.1 mM to about 50 mM, 1.0 mM to about 900 mM, 1.0 mM to about 500 mM, 1.0 mM to about 100 mM, 1.0 mM to about 90 mM, or 1.0 mM to about 50 mM. The concentration of the buffer, most preferably histidine, is preferably 20 mM.

[0079] Embodiments of the present invention can be practiced with compositions comprising multiple botulinum toxin serotypes, such as a botulinum toxin serotype selected from the group consisting of botulinum toxin serotypes A, B, C1, D, E, F, G, and mosaic. In certain embodiments, purified botulinum toxin can be used. In other embodiments, modified botulinum toxin can be used.

[0080] In some embodiments, a clostridial toxin pharmaceutical composition of the invention can be formulated as a lyophilized (i.e., freeze-dried) or vacuum-dried powder that can be reconstituted with a suitable fluid, such as saline or water, prior to administration to a patient. In another embodiment of the invention, the pharmaceutical composition can be formulated as an aqueous solution or suspension.

[0081] In some embodiments, a solid clostridial toxin pharmaceutical composition of the invention comprises a botulinum toxin, an isotonicity agent, a poloxamer and / or polysorbate, and an antioxidant. In some embodiments, the clostridial toxin pharmaceutical composition comprises a botulinum toxin. In some embodiments, the clostridial toxin pharmaceutical composition comprises trehalose. In some embodiments, the clostridial toxin pharmaceutical composition comprises poloxamer 188 or polysorbate 20. In some embodiments, the composition comprises EDTA, EGTA, DTPA, or an analog thereof. In alternative embodiments, the composition comprises methionine and / or NAC. In aspects of these alternative embodiments, the composition further comprises EDTA, EGTA, DTPA, or an analog thereof. In some embodiments, the composition further comprises a buffering agent. In one embodiment, the composition comprises a histidine buffering agent. In some embodiments, the relative weight amounts of trehalose, poloxamer, and methionine are within the ranges of 1-10%, 0.5-5%, and 0.1-0.3%, respectively. In some embodiments, the relative weight amounts of trehalose, polysorbate, and methionine are within the ranges of 1-10%, 0.02%-0.06%, and 0.1-0.3%, respectively. In some embodiments, the relative weight amount of EDTA or an EDTA analog is about 0.01-0.10%. In some embodiments, the relative weight amount of NAC is within the range of 0.01-0.5%.

[0082] In aspects of these embodiments, the clostridial toxin pharmaceutical composition is formulated as a solid (i.e., lyophilized or vacuum-dried) composition. In some embodiments, the solid clostridial pharmaceutical composition comprises a lyoprotectant. In some embodiments, preferred lyoprotectants include sucrose, trehalose, mannitol, sorbitol, or combinations thereof. In some embodiments, the solid pharmaceutical composition comprises NAC at a relative weight ratio of 0.01-0.05%. In some embodiments, the pharmaceutical composition further comprises EDTA, EGTA, DTPA, or analogs thereof. In alternative embodiments, the solid pharmaceutical composition comprises methionine and EDTA or an EDTA analog.

[0083] In alternative aspects of these embodiments, the clostridial toxin pharmaceutical composition is formulated as a liquid. In some embodiments, the liquid pharmaceutical composition comprises NAC at a relative weight ratio of 0.1-0.5%. In some embodiments, the liquid pharmaceutical composition comprises NAC and EDTA, EGTA, DTPA, or analogs thereof. In some embodiments, the liquid pharmaceutical composition comprises a histidine buffer. In some embodiments, the liquid pharmaceutical composition has a pH of 5-7.

[0084] The present invention provides (i) a botulinum toxin; (ii) poloxamer 188, (iii) methionine or N-acetyl-cysteine.

[0085] The combination of poloxamer 188 with either methionine or N-acetylcysteine of the present invention is preferred due to the synergistic effect on the stability of the composition caused by the combination of this particular surfactant with either of these antioxidants.

[0086] The composition may further comprise trehalose or sucrose, with trehalose being preferred over sucrose for solid compositions.

[0087] In addition, the composition may optionally contain NaCl. NaCl may be particularly preferably contained in a composition comprising a botulinum toxin, trehalose or sucrose, poloxamer 188, and methionine, and particularly preferably contained in a liquid composition comprising a botulinum toxin, trehalose or sucrose, poloxamer 188, and methionine. In some lyophilized formulations, NaCl may function as a tonicity agent in the reconstitution medium.

[0088] In addition to or instead of NaCl, the composition may optionally contain EDTA, EGTA, DTPA, or analogs thereof. EDTA may be included in compositions containing botulinum toxin, trehalose or sucrose, poloxamer 188, and methionine, and is preferably included in liquid compositions containing botulinum toxin, trehalose or sucrose, poloxamer 188, and methionine.

[0089] EDTA, EGTA, DTPA, or analogs thereof may also be included in compositions containing botulinum toxin, trehalose or sucrose, poloxamer 188, and NAC, and are preferably included in liquid compositions containing botulinum toxin, trehalose or sucrose, poloxamer 188, and NAC. The combination of EDTA and NAC is particularly preferred because it has a synergistic stabilizing effect on the compositions of the present invention.

[0090] The relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, methionine, and NaCl (if present), and EDTA (if present) are preferably within the following ranges: [Table 1] Such compositions may be both liquid and solid formulations. When the composition is a solid composition, the sugar is preferably trehalose.

[0091] In addition, these compositions preferably contain a buffer. In the composition of the present invention, when the composition is a liquid composition, the buffer is preferably histidine, and the concentration of histidine is preferably 20 mM. In these compositions, the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, and most preferably within the range of 5.5 to 6. On the other hand, when the composition is solid, the histidine is preferably contained in a relative weight amount of 0.1 to 0.5 (% w / w), more preferably within the range of 0.3-0.4% (w / w).

[0092] More preferably, the relative weight percentages (w / w) of trehalose, poloxamer 188, and methionine are as follows: [Table 2]

[0093] In the present invention, the composition designated as a) above may be a solid composition, preferably lyophilized. This composition also preferably contains a buffer, preferably histidine, which is preferably present in the solid composition in a relative weight ratio of 0.1 to 0.5% (w / w), more preferably 0.3 to 0.4% (w / w). Histidine may be present as a pure substance, a histidine acid salt (e.g., histidine HCl), a histidine acid salt hydrate, or a mixture of any of the foregoing. In a preferred embodiment, histidine base may be present in an amount of 1.43 mg / mL (or 0.14% w / w), together with histidine hydrochloride in an amount of 2.26 mg / mL (or 0.23% w / w). Thus, the total concentration of histidine species (total histidine concentration) is 0.37% w / w. In one embodiment, the solid composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient.

[0094] The solid composition most preferably has one of the following compositions: [Table 3]

[0095] In one embodiment of the present invention, the composition is a solid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, methionine, and a buffering agent, preferably present in the amounts described above. In one embodiment, the solid composition is reconstituted with a reconstitution medium comprising NaCl prior to administration to a patient. In one embodiment, NaCl may be present in the reconstitution medium in an amount of 0.9% (w / w).

[0096] In the present invention, the composition indicated as a) above may be a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6.

[0097] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, methionine, and a buffering agent, these components preferably being present in the amounts described above.

[0098] Even more preferably, the relative weight percentages (w / w) of trehalose, poloxamer 188, and methionine are as follows: [Table 4]

[0099] In the present invention, the composition designated as b) above is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 5.5.

[0100] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, methionine, and a buffering agent, these components preferably being present in the amounts described above.

[0101] Even more preferably, the relative weight amounts (%, w / w) of trehalose, poloxamer 188, methionine, and NaCl are as follows: [Table 5]

[0102] In the present invention, the composition designated as c) above is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 5.5.

[0103] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, methionine, NaCl, and a buffering agent, these components preferably being present in the amounts described above.

[0104] Even more preferably, the relative weight percentages (w / w) of trehalose, poloxamer 188, methionine, and EDTA are as follows: [Table 6]

[0105] In the present invention, the composition designated as d) above is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6.

[0106] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, methionine, EDTA, and a buffer, these components preferably being present in the amounts described above.

[0107] The relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, N-acetyl-cysteine, and EDTA (if present) are preferably within the following ranges: [Table 7]

[0108] Such compositions may be both liquid and solid formulations, preferably liquid compositions. When the composition is a solid composition, the sugar is preferably trehalose.

[0109] In addition, these compositions preferably contain a buffer. In the composition of the present invention, when the composition is a liquid composition, the buffer is preferably histidine, and the concentration of histidine is preferably 20 mM. In these compositions, the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, and most preferably within the range of 5.5 to 6. On the other hand, when the composition is solid, the histidine is preferably contained in a relative weight amount of 0.1 to 0.5 (% w / w), more preferably within the range of 0.3-0.4% (w / w).

[0110] More preferably, the relative weight amounts (%, w / w) of trehalose, poloxamer 188, N-acetyl-cysteine, and EDTA are as follows: [Table 8] In the present invention, the composition is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6.

[0111] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, N-acetyl-cysteine, EDTA, and a buffer, these components preferably being present in the amounts described above.

[0112] As noted above, EDTA is an optional ingredient in the N-acetyl-cysteine-containing compositions of the present invention. In such EDTA-free compositions, the relative weight percentages (w / w) of trehalose, poloxamer 188, and N-acetyl-cysteine may be as follows: [Table 9]

[0113] Alternatively, the relative weights (%, w / w) of the ingredients in the EDTA-free composition are as follows: [Table 10]

[0114] In the present invention, the EDTA-free composition is preferably lyophilized and preferably a solid composition. The composition also preferably contains a buffer, preferably histidine, which is preferably present in the solid composition at a relative weight ratio of 0.1 to 0.5% (w / w), more preferably 0.3 to 0.4% (w / w). Histidine may be present as a pure substance, a histidine acid salt, a histidine acid salt hydrate, or a mixture of any of the foregoing. In a preferred embodiment, histidine base may be present in an amount of 1.43 mg / mL (or 0.14% w / w), together with histidine hydrochloride in an amount of 2.26 mg / mL (or 0.23% w / w). Thus, the total concentration of histidine species (total histidine concentration) is 0.37% w / w.

[0115] In one embodiment of the present invention, the composition is a solid composition comprising one or more botulinum toxins, trehalose, poloxamer 188, N-acetyl-cysteine, and a buffering agent, these components preferably being present in the amounts described above.

[0116] In some cases, the compositions of the present invention do not contain sugars or polyalcohols. As a result, the compositions contain botulinum toxin, poloxamer 188, and methionine or N-acetyl-cysteine, but do not contain sugars or polyalcohols. When the composition contains N-acetyl-cysteine, it is preferred that it also contains EDTA.

[0117] In such compositions, the relative weight percentages (w / w) of poloxamer 188 and methionine are preferably within the following ranges: [Table 11]

[0118] On the other hand, the relative weight ratios (%, w / w) of poloxamer 188, and N-acetyl-cysteine and EDTA are preferably within the following ranges: [Table 12]

[0119] Such compositions are preferably liquid formulations.

[0120] More preferably, the relative weights (%, w / w) of poloxamer 188 and methionine are as follows: [Table 13]

[0121] As mentioned above, such compositions are preferably liquid compositions. In this case, the composition preferably contains a buffer, preferably histidine. The histidine concentration is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6. These compositions may also contain NaCl. NaCl may be present in a relative weight amount of 0.5 to 1.5% (w / w), more preferably 0.9% (w / w).

[0122] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, poloxamer 188, methionine, and a buffering agent, and optionally NaCl, with these components preferably present in the amounts described above.

[0123] Further preferred are the following relative weight amounts (%, w / w) of poloxamer 188 and N-acetyl-cysteine: [Table 14]

[0124] Such compositions are preferably liquid compositions. In this case, the composition preferably contains a buffer, preferably histidine. The histidine concentration is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6. These compositions may also contain NaCl. NaCl may be present in a relative weight amount of 0.5 to 1.5% (w / w), more preferably 0.9% (w / w).

[0125] In one embodiment of the invention, the composition is a liquid composition comprising one or more botulinum toxins, poloxamer 188, N-acetyl-cysteine, a buffer, and optionally EDTA, and optionally NaCl, with these components preferably present in the amounts described above.

[0126] In another embodiment, the sugar- or polyalcohol-free composition may contain Tween-20 instead of poloxamer-188. Such a composition may contain Tween-20 in an amount of 0.01 to 0.1% (w / w), more preferably 0.02 to 0.06% (w / w), and most preferably 0.04% (w / w).

[0127] In another preferred embodiment of the present invention, the relative weight amounts (% w / w) of sucrose, poloxamer 188, and methionine are as follows: [Table 15]

[0128] In the present invention, the composition designated as a') above may preferably be a lyophilized solid composition. This composition also preferably contains a buffer, preferably histidine, which is preferably present in the solid composition in a relative weight ratio of 0.1 to 0.5% (w / w), more preferably 0.3 to 0.4% (w / w). Histidine may be present as a pure substance, a histidine acid salt (e.g., histidine HCl), a histidine acid salt hydrate, or a mixture of any of the foregoing. In a preferred embodiment, histidine base may be present in an amount of 1.43 mg / mL (or 0.14% w / w), together with histidine hydrochloride in an amount of 2.26 mg / mL (or 0.23% w / w). Thus, the total concentration of histidine species (total histidine concentration) is 0.37% w / w.

[0129] The solid composition most preferably has one of the following compositions: [Table 16]

[0130] In one embodiment of the present invention, the composition is a solid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, methionine, and a buffering agent, preferably present in the amounts described above. In one embodiment, the solid composition is reconstituted with a reconstitution medium comprising NaCl prior to administration to a patient. In one embodiment, NaCl may be present in the reconstitution medium in an amount of 0.9% (w / w).

[0131] In the present invention, the composition indicated as a') above may be a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6.

[0132] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, methionine, and a buffering agent, these components preferably being present in the amounts described above.

[0133] Even more preferably, the relative weight percentages (w / w) of sucrose, poloxamer 188, and methionine are as follows: [Table 17]

[0134] In the present invention, the composition designated as b') above is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 5.5.

[0135] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, methionine, and a buffering agent, these components preferably being present in the amounts described above.

[0136] Even more preferably, the relative weight percentages (w / w) of sucrose, poloxamer 188, methionine, and NaCl are as follows: [Table 18]

[0137] In the present invention, the composition designated as c') above is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 5.5.

[0138] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, methionine, NaCl, and a buffer, these components preferably being present in the amounts described above.

[0139] Even more preferably, the relative weight percentages (w / w) of sucrose, poloxamer 188, methionine, and EDTA are as follows: [Table 19]

[0140] In the present invention, the composition indicated as d') above is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, more preferably within the range of 5.5 to 6, and most preferably 6.

[0141] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, methionine, EDTA, and a buffer, these components preferably being present in the amounts described above.

[0142] In another preferred embodiment, the relative weight amounts (%, w / w) of sucrose, poloxamer 188, N-acetyl-cysteine, and EDTA are as follows: [Table 20]

[0143] In the present invention, the composition is preferably a liquid composition. In this case, the composition preferably contains a buffer, and the buffer is preferably histidine. The concentration of histidine is preferably 20 mM, and the pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, even more preferably within the range of 5.5 to 6, and most preferably 6.

[0144] In one embodiment of the present invention, the composition is a liquid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, N-acetyl-cysteine, EDTA, and a buffer, these components preferably being present in the amounts described above.

[0145] As noted above, EDTA is an optional ingredient in the N-acetyl-cysteine-containing compositions of the present invention. In such EDTA-free compositions, the relative weight percentages (w / w) of sucrose, poloxamer 188, and N-acetyl-cysteine may be as follows: [Table 21]

[0146] Alternatively, the relative weights (%, w / w) of the ingredients in the EDTA-free composition are as follows: [Table 22]

[0147] In the present invention, the EDTA-free composition is preferably lyophilized and preferably a solid composition. The composition also preferably contains a buffer, preferably histidine, which is preferably present in the solid composition at a relative weight ratio of 0.1 to 0.5% (w / w), more preferably 0.3 to 0.4% (w / w). Histidine may be present as a pure substance, a histidine acid salt, a histidine acid salt hydrate, or a mixture of any of the foregoing. In a preferred embodiment, histidine base may be present in an amount of 1.43 mg / mL (or 0.14% w / w), together with histidine hydrochloride in an amount of 2.26 mg / mL (or 0.23% w / w). Thus, the total concentration of histidine species (total histidine concentration) is 0.37% w / w.

[0148] In one embodiment of the present invention, the composition is a solid composition comprising one or more botulinum toxins, sucrose, poloxamer 188, N-acetyl-cysteine, and a buffering agent, these components preferably being present in the amounts described above.

[0149] The composition of the present invention may be a liquid composition, in which case the composition preferably contains a buffer. The buffer is preferably histidine, and the concentration of histidine is preferably 20 mM. The pH is preferably within the range of 5 to 7, more preferably within the range of 5.5 to 6.5, and most preferably within the range of 5.5 to 6.

[0150] The composition of the present invention may also be a solid composition, preferably lyophilized. This composition may further contain a buffering agent, preferably histidine, in a relative weight ratio of 0.1 to 0.5% (w / w), more preferably 0.3 to 0.4% (w / w).

[0151] It is particularly preferred that the compositions of the present invention are free of animal-derived proteins.

[0152] Any of the aforementioned compositions containing botulinum toxin, poloxamer 188, and methionine or N-acetyl-cysteine may further contain ethylenediaminetetraacetic acid sodium salt (EDTA) or an EDTA analog. The relative weight (% w / w) of EDTA in these compositions may be in the range of about 0.01 to 0.10.

[0153] In the present invention, the term "antioxidant" refers to any compound that protects an active ingredient from reacting with oxygen. Antioxidants can be broadly divided into three categories: (i) sacrificial antioxidants that can scavenge oxygen by reacting with it more readily than a particular active ingredient. In the present invention, this type of antioxidant is preferably ascorbic acid; (ii) Chain terminators that react with free radicals and peroxides to prevent the propagation of radical chains. In the present invention, methionine, cysteine, N-acetyl-cysteine, and BHT are preferred chain terminators; (iii) A chelating agent that reduces the catalytic activity of a transition metal by forming a complex with the metal. In the present invention, EDTA, EGTA, and DTPA are preferred chelating agents.

[0154] The following compositions define the present invention by reference to the general or specific types of antioxidants discussed in the paragraph above. All of these compositions may optionally contain a buffer. In the following liquid compositions, the buffer is preferably histidine, the concentration of histidine is preferably 20 mM, the pH is preferably in the range of 5 to 7, even more preferably in the range of 5.5 to 6.5, and most preferably in the range of 5.5 to 6. In the following solid compositions, the buffer is preferably histidine, and the histidine is preferably present in a relative weight amount of 0.1 to 0.5% (w / w), more preferably in a relative weight amount of 0.3 to 0.4% (w / w). In addition, all of the following compositions may contain NaCl.

[0155] The present invention also provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a poloxamer, (iii) a chelating agent; and (iv) a sacrificial antioxidant.

[0156] In a preferred embodiment of this liquid pharmaceutical composition, the chelating agent is EDTA, EGTA, or DTPA, and the sacrificial antioxidant is ascorbic acid. In such a composition, the clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188.

[0157] The present invention also provides a liquid pharmaceutical composition comprising: (i) clostridial toxins, (ii) poloxamer, (iii) a chelating agent, and / or (iv) a chain terminator.

[0158] In a preferred embodiment of this liquid pharmaceutical composition, the chelating agent is EDTA and the chain terminator is N-acetyl-cysteine. In another preferred embodiment, the chelating agent is EDTA and the chain terminator is butylated hydroxytoluene (BHT). In another preferred embodiment, the chelating agent is EDTA and the chain terminator is methionine. In another preferred embodiment of this liquid pharmaceutical composition, the chelating agent is DTPA and the chain terminator is N-acetyl-cysteine. In another preferred embodiment, the chelating agent is DTPA and the chain terminator is BHT. In another preferred embodiment, the chelating agent is DTPA and the chain terminator is methionine. In another preferred embodiment of this liquid pharmaceutical composition, the chelating agent is EGTA and the chain terminator is N-acetyl-cysteine. In another preferred embodiment, the chelating agent is EGTA and the chain terminator is BHT. In another preferred embodiment, the chelating agent is EGTA and the chain terminator is methionine. In another preferred embodiment, the liquid composition comprises a chain terminator rather than a chelating agent, wherein the chain terminator is NAC. In all of these liquid compositions, the Clostridial toxin is preferably a botulinum toxin and the poloxamer is preferably poloxamer-188.

[0159] The present invention also provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a poloxamer, (iii) methionine.

[0160] In this embodiment, the Clostridial toxin is preferably a botulinum toxin and the poloxamer is preferably poloxamer-188. In such compositions, the poloxamer-188 and methionine are preferably present in the amounts defined above for compositions that do not contain sugars or polyalcohols.

[0161] The present invention also provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof; (ii) a poloxamer, (iii) methionine.

[0162] In this embodiment, the Clostridial toxin is preferably a botulinum toxin and the poloxamer is preferably poloxamer-188. In such compositions, the poloxamer-188 and methionine are preferably present in the amounts defined above for compositions comprising a sugar or a polyalcohol.

[0163] The present invention also preferably provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a sacrificial antioxidant.

[0164] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, the sacrificial antioxidant is preferably ascorbic acid, the clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80.

[0165] The present invention also preferably provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a chain terminator.

[0166] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, the chain terminator is preferably NAC, the clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80.

[0167] The present invention also preferably provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, NaCl, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a chain terminator.

[0168] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, the chain terminator is preferably NAC, the clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80.

[0169] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a lyoprotectant selected from trehalose, sucrose, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a sacrificial antioxidant.

[0170] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, and the sacrificial antioxidant is preferably ascorbic acid. The Clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80. In particular embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0171] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a lyoprotectant selected from trehalose, sucrose, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a chain terminator.

[0172] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, and the chain terminator is preferably NAC. The clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80. In particular embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0173] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a lyoprotectant selected from trehalose, sucrose, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chain terminator.

[0174] In this particular embodiment, the chain terminator is preferably NAC or methionine. The Clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80. In particular embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0175] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, or a combination thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a chain terminator.

[0176] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, and the chain terminator is preferably NAC. The clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80. In particular embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0177] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, (iv) a chelating agent; and (v) a sacrificial antioxidant.

[0178] In this particular embodiment, the chelating agent is preferably EDTA, EGTA, or DTPA, and the sacrificial antioxidant is preferably ascorbic acid. The Clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188. In particular embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0179] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, (iv) a chelating agent; and (v) a chain terminator.

[0180] In a preferred embodiment, the chelating agent is EDTA and the chain terminator is N-acetyl-cysteine. The Clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188. In certain embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0181] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, (iv) a chain terminator.

[0182] In a preferred embodiment, the chain terminator is N-acetyl-cysteine. The Clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188. In certain embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0183] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, (iv) methionine.

[0184] In preferred embodiments, the Clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188. In certain embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0185] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a lyoprotectant selected from trehalose, sucrose, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) at least two antioxidants selected from the list of chelating agents, chain terminators, and sacrificial antioxidants.

[0186] In this aspect of the invention, the clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80. The chelating agent is preferably EDTA, EGTA, or DTPA, the sacrificial antioxidant is preferably ascorbic acid, and the chain terminator is preferably methionine, cysteine, NAC, or BHT. Consequently, the two antioxidants preferably include a combination of ascorbic acid and methionine, cysteine, NAC, or BHT. The two antioxidants also preferably include a combination of ascorbic acid and EDTA, EGTA, or DTPA. The two antioxidants also preferably include a combination of EDTA and methionine, cysteine, NAC, or BHT. The two antioxidants also preferably include a combination of DTPA and methionine, cysteine, NAC, or BHT. The two antioxidants may also preferably be a combination of EGTA and methionine, cysteine, NAC, or BHT. In certain embodiments, the lyophilized composition is reconstituted with a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof. In at least one embodiment, the lyophilized composition is reconstituted with a reconstitution medium containing NaCl before administration to a patient. In at least one embodiment, NaCl is present in the reconstitution medium in an amount of 0.9% (w / w).

[0187] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, or glucose; (iii) a poloxamer, (iv) a chelating agent; and (v) a chain terminator.

[0188] In a preferred embodiment, the chelating agent is EDTA, the chain terminator is N-acetyl-cysteine or methionine, the clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188.

[0189] The present invention also provides a lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, or glucose; (iii) a poloxamer, (v) a chain terminator.

[0190] In a preferred embodiment, the chain terminator is N-acetyl-cysteine or methionine. The clostridial toxin is preferably a botulinum toxin, and the poloxamer is preferably poloxamer-188.

[0191] The present invention also provides a liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) at least two antioxidants selected from the list of chelating agents, chain terminators, and sacrificial antioxidants.

[0192] In this aspect of the invention, the clostridial toxin is preferably a botulinum toxin, the poloxamer is preferably poloxamer-188, and the polysorbate is preferably polysorbate-20 or 80. The chelating agent is preferably EDTA, EGTA, or DTPA, the sacrificial antioxidant is preferably ascorbic acid, and the chain terminator is preferably methionine, cysteine, NAC, or BHT. Consequently, the two antioxidants preferably include a combination of ascorbic acid and methionine, cysteine, NAC, or BHT. The two antioxidants also preferably include a combination of ascorbic acid and EDTA, EGTA, or DTPA. The two antioxidants also preferably include a combination of EDTA and methionine, cysteine, NAC, or BHT. The two antioxidants also preferably include a combination of DTPA and methionine, cysteine, NAC, or BHT. The two antioxidants also preferably include a combination of EGTA with methionine, cysteine, NAC, or BHT. Treatment method

[0193] In embodiments, the present invention provides methods comprising administering to a subject in need of treatment for a disease, disorder, condition, etc., a pharmaceutical formulation of the present invention in an amount sufficient to produce improved patient function. In certain embodiments, the disease is neuromuscular in nature, e.g., a disease affecting muscles and their neural control, such as overactive bladder. In preferred methods, the compositions of the present invention are used to treat cardiac arrhythmias. Certain embodiments relate to the treatment of pain, e.g., the treatment of headaches, or back pain, or muscle pain, etc. In certain embodiments, the methods of the present invention encompass the treatment of psychiatric disorders, including, e.g., depression, anxiety, etc. In preferred methods, the compositions of the present invention are used to treat depression.

[0194] When the composition is used to treat depression, it is preferably used to treat adults with either single episode or recurrent moderate to severe major depressive disorder (MDD), a diagnosis based on the Diagnostic and Statistical Manual of Mental Disorders, Text Edition (DSM-IV-TR), published in 2000. While any of the compositions of the invention can be used, it is preferred to use a lyophilized botulinum toxin type A formulation described in Table A, formulated as a solution for administration. [Table 23]

[0195] In one embodiment, the composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In one embodiment, NaCl may be present in the reconstitution medium in an amount of 0.9% (w / w).

[0196] The composition is preferably administered in a single treatment session. The dosage is preferably a total dose of 30 or 50 units. These doses are preferably injected into six (for a 30-unit dose) or eight (for a 50-unit dose) injection sites in the glabellar region of the forehead, preferably in the procerus and corrugator supercilii muscles. This treatment can be repeated at 3-6 month intervals. One advantage of using this method is that patients treated with this composition show improvement in the clinical Montgomery Asberg Depression Rating Scale. In addition, these patients also experience improvements in clinical CGI-S scores (Clinical Global Impression Change Score) and clinical HAM-D17 total scores (Hamilton Rating Scale for Depression).

[0197] The compositions and methods of the present invention are useful for treating conditions such as achalasia, anal fissures, anismus, blepharospasm, cerebral palsy, cervical dystonia, cervicogenic headache, hemifacial spasm, dyshidrotic eczema, dysphagia, dysphonia, esophageal hypoperistalsis, orbicularis esophagealis, convergence strabismus (infants), eye lift, facial spasms, gait disorders (idiopathic tiptoe gait), generalized dystonia, hemifacial spasm, excessive facial wrinkles (glabellar, forehead, crow's feet, marionette lines), hyperhidrosis, incontinence (idiopathic or neurogenic), drug-induced headache, headache, migraine, myoclonus, including those involving the masseter muscle. muscle mass or activity reduction due to steroids, myofascial pain syndrome, obstructive urinary symptoms, pancreatic duct nonunion pancreatitis, Parkinson's disease, puborectalis syndrome, decreased surgical scar pressure, excessive salivation, salivary gland myxoma, 6th cranial nerve palsy, spasticity, speech / voice disorders, strabismus, surgical assistance (ophthalmology), tardive dyskinesia, temporomandibular joint disorders, tension headache, thoracic outlet syndrome, torsion dystonia, torticollis, Tourette's syndrome, tremor, whiplash-related neck pain, pain, pruritus, inflammation, allergies, cancer and benign tumors, fever, obesity, infections (viral and bacterial), hypertension, cardiac arrhythmia, vasospasm, atheroma Arteriosclerosis, endothelial hyperplasia, venous thrombosis, varicose veins, aphthous stomatitis, hypersalivation, temporomandibular joint syndrome, hyperhidrosis, bromhidrosis, acne, rosacea, hyperpigmentation, hypertrophic scars, keloids, calluses and corns, skin wrinkles, excessive sebum production, psoriasis, dermatitis, allergic rhinitis, nasal congestion, postnasal drip, sneezing, earwax, exudative and suppurative otitis media, tonsillar and adenoid hypertrophy, tinnitus, dizziness, vertigo, hoarseness, cough, sleep apnea, snoring, glaucoma, conjunctivitis, uveitis, strabismus, Graves' disease, hirsutism, hair loss, asthma, bronchitis, emphysema, mucus production, pleural The compositions of the present invention may be useful for treating, reducing, and / or preventing the symptoms of: inflammation, coagulation disorders, myeloproliferative disorders, disorders involving eosinophils, neutrophils, macrophages, and lymphocytes, immune tolerance and transplantation, autoimmune disorders, dysphagia, acid reflux, hiatal hernia, gastritis and hyperacidity, diarrhea and constipation, hemorrhoidal disorders, urinary incontinence, prostatic hyperplasia, erectile dysfunction, priapism and Peyronie's disease, epididymitis, contraception, menstrual cramps, prevention of premature labor, endometriosis and fibrosis, arthritis, osteoarthritis, rheumatism, bursitis, tendonitis, tenosynovitis, fibromyalgia, seizure disorders, spasticity, headache, and neuralgia. In a preferred method, the compositions of the present invention are used to treat cardiac arrhythmias.

[0198] When the composition is used to treat cardiac arrhythmia, the patient is typically undergoing cardiac surgery. The formulation used for treatment may be any of the compositions of the present invention. However, the formulation is preferably based on the lyophilized formulation described in Table A, which is incorporated into a solution for administration. [Table 24]

[0199] In one embodiment, the composition is reconstituted with a reconstitution medium containing NaCl prior to administration to a patient. In one embodiment, NaCl may be present in the reconstitution medium in an amount of 0.9% (w / w).

[0200] The composition is administered via a single injection into the major epicardial fat pad of the heart. The dose used is 25 U per epicardial fat pad (total dose of 125 U) or 50 U per epicardial fat pad (total dose of 250 U). One advantage of using this method is a reduction in the incidence of atrial fibrillation (AF) as measured by ECG at 4 weeks. Other advantages include a reduction in hospital length of stay, a reduction in ICU length of stay, a reduction in patient readmission rates, a reduction in the use of anticoagulants, and a reduction in the need for interventional procedures for postoperative atrial fibrillation (POAF), such as ablation, pacemaker implantation, or electrical or pharmacological cardioversion.

[0201] In certain embodiments, the maximum dosage administered to a patient is limited to a maximum of 360 U of botulinum toxin in any 90-day period. Treatment of neurological / muscular symptoms

[0202] In one embodiment, the neuromuscular disorder is hyperhidrosis. For example, a subject suffering from hyperhidrosis may be treated with the pharmaceutical formulation of the present invention at a dose of about 59 U per axilla, about 58 U per axilla, about 57 U per axilla, about 56 U per axilla, about 55 U per axilla, about 54 U per axilla, about 53 U per axilla, about 52 U per axilla, about 51 U per axilla, about 50 U per axilla, or about 49 U per axilla, or about 54 U per axilla, or about 53 U per axilla, or about 52 U per axilla, or about 51 U per axilla, or about 50 U per axilla, or about 49 U per axilla, or about 54 ... In one embodiment, a total of 50 U is administered intradermally at 10-15 sites spaced approximately 1-2 cm apart.

[0203] In one embodiment, the neuromuscular disorder is hemifacial spasm. For example, a subject suffering from hemifacial spasm is administered about 1.5 to 15 U of the pharmaceutical formulation of the present invention per treatment. In a further example, a subject suffering from hemifacial spasm is administered about 1.5 to 3 U, 1.5 to 5 U, 1.5 to 7 U, 1.5 to 10 U, 1.5 to 12 U, 1.5 to 15 U, 5 to 10 U, 5 to 15 U, or 10 to 15 U per treatment. In yet a further example, a subject is administered about 1.5U, about 2U, about 2.5U, about 3U, about 3.5U, about 4U, about 4.5U, about 5U, about 5.5U, about 6U, about 6.5U, about 7U, about 7.5U, about 8U, about 8.5U, about 9U, about 9.5U, about 10U, about 10.5U, about 11U, about 11.5U, about 12U, about 12.5U, about 13U, about 13.5U, about 14U, about 14.5U, or about 15U per treatment to a patient with hemifacial spasm. Doses of more than 15U per treatment can also be administered to a patient with hemifacial spasm to achieve a therapeutic response. A treatment session can include multiple treatments.

[0204] In one embodiment, the neuromuscular disorder is cervical dystonia. For example, a subject suffering from cervical dystonia is administered approximately 1.5 to 300 U of the pharmaceutical formulation of the present invention per treatment. In a further example, a subject with cervical dystonia is administered approximately 35 to 250 U, 65 to 200 U, 85 to 175 U, 105 to 160 U, or 125 to 145 U. In one embodiment, the dosage administered to the sternocleidomastoid muscle is limited to 100 U or less. Doses exceeding 300 U per treatment can also be administered to patients with cervical dystonia to achieve a therapeutic response. A treatment session can include multiple treatments.

[0205] In one embodiment, the neuromuscular disorder is blepharospasm. A subject suffering from blepharospasm is administered, for example, about 1.25 to 2.5 U of a pharmaceutical formulation of the present invention, which is injected into the medial and lateral pretarsal orbicularis oculi muscles of the upper eyelid and the lateral pretarsal orbicularis oculi muscle of the lower eyelid. In a further example, a subject is administered about 1.5 U, about 1.6 U, about 1.7 U, about 1.8 U, about 1.9 U, about 2.0 U, about 2.1 U, about 2.2 U, about 2.3 U, about 2.4 U, about 2.5 U, or more per injection site. A treatment session may include multiple treatments.

[0206] In one embodiment, the neuromuscular disorder is strabismus. A subject suffering from strabismus is administered, for example, about 1.25 to 2.5 U of the pharmaceutical formulation of the present invention per injection site. In a further example, a subject is administered about 1.5 U, about 1.6 U, about 1.7 U, about 1.8 U, about 1.9 U, about 2.0 U, about 2.1 U, about 2.2 U, about 2.3 U, about 2.4 U, about 2.5 U, or more per injection site to achieve a therapeutic response. In embodiments, lower doses are used to treat minor anomalies. In embodiments, vertical muscle and horizontal strabismus of less than 20 prism diameters can be treated with 1.25 to 2.5 U per injection site. A treatment session may include multiple treatments.

[0207] In one embodiment, the neuromuscular disorder is muscle spasticity. A subject suffering from muscle spasticity is administered, for example, about 20-200 U per treatment of the pharmaceutical formulation of the present invention. In a further example, a subject suffering from muscle spasticity is administered about 20-30 U, 20-40 U, 20-60 U, 20-80 U, 20-100 U, 20-125 U, 20-150 U, or 20-175 U per treatment. In yet a further example, a subject is administered about 20 U, about 25 U, about 30 U, about 35 U, about 40 U, about 45 U, about 50 U, about 55 U, about 60 U, about 65 U, about 70 U, about 75 U, about 80 U, about 85 U, about 90 U, about 95 U, about 100 U, about 105 U, about 110 U, about 115 U, about 120 U, about 125 U, about 130 U, about 135 U, about 140 U, about 145 U, about 150 U, about 155 U, about 160 U, about 165 U, about 170 U, about 175 U, about 180 U, about 185 U, about 190 U, about 195 U, or about 200 U per treatment to a patient with muscle spasticity. In some embodiments, the biceps brachii muscle may receive 100 U to 200 U divided into four injection sites. In some embodiments, the flexor carpi radialis muscle may receive 12.5 U to 50 U in one injection site. In some embodiments, the flexor carpi ulnaris muscle may receive 12.5 U to 50 U in one injection site. In some embodiments, the flexor digitorum profundus muscle may receive 30 U to 50 U in one injection site. In some embodiments, the flexor digitorum superficialis muscle may receive 30 U to 50 U in one injection site. Doses greater than 200 U per treatment may also be administered to patients with muscle spasticity to achieve a therapeutic response. A treatment session may include multiple treatments. Treating pain

[0208] In another embodiment, the present invention provides a method of treating pain, comprising administering to a subject in need thereof a pharmaceutical formulation of the present invention in an amount sufficient to reduce pain. In another embodiment, the patient is suffering from myofascial pain, migraine, tension headache, neuropathic pain, facial pain, lower back pain, sinus headache, pain associated with temporomandibular joint disorder, pain associated with spasticity or cervical dystonia, post-surgical wound pain, or neuralgia. A treatment session may include multiple treatments.

[0209] In some embodiments, the patient is suffering from facial pain. For example, a subject suffering from facial pain is administered about 4-40 U per treatment of a pharmaceutical formulation of the present invention. In further examples, a subject suffering from facial pain is administered about 4-10 U, 4-15 U, 4-20 U, 4-25 U, 4-30 U, 4-35 U, 7-15 U, 7-20 U, 7-25 U, 7-30 U, 7-35 U, or 7-40 U per treatment. In yet a further example, a subject is administered about 4 U, about 5 U, about 7.5 U, about 10 U, about 12.5 U, about 15 U, about 17.5 U, about 20.0 U, about 22.5 U, about 25.0 U, about 27.5 U, about 30.0 U, about 32.5 U, about 35 U, about 37.5 U, or about 40 U per treatment to a facial pain patient. Doses greater than 40 U per treatment can also be administered to a facial pain patient to achieve a therapeutic response. A treatment session can include multiple treatments.

[0210] In one embodiment, the patient is suffering from myofascial pain. A subject suffering from myofascial pain is administered, for example, about 5-100 U per treatment of a pharmaceutical formulation of the present invention. In a further example, a subject suffering from myofascial pain is administered about 5-10 U, 5-20 U, 5-30 U, 5-40 Units, 5-50 Units, 5-60 Units, 5-70 Units, 5-80 Units, 5-90 U, 10-20 U, 10-30 U, 10-50 U, 10-60 U, 10-70 U, 10-80 U, 10-90 U, or 10-100 U per treatment. In a further example, a subject is administered about 5 U, about 10 U, about 15 U, about 20 U, about 25 U, about 30 U, about 35 U, about 40 U, about 45 U, about 50 U, about 55 U, about 60 U, about 65 U, about 70 U, about 75 U, about 80 U, about 85 U, about 90 U, about 95 U, or about 100 U per treatment to a myofascial pain patient. Doses greater than 100 U per treatment can also be administered to a myofascial pain patient to achieve a therapeutic response. A treatment session can include multiple treatments.

[0211] In one embodiment, the patient is suffering from lower back pain. For example, a subject suffering from lower back pain is administered about 15-150 U per treatment of a pharmaceutical formulation of the present invention. In a further example, a subject suffering from lower back pain is administered about 15-30 U, 15-50 U, 15-75 U, 15-100 U, 15-125 U, 15-150 U, 20-100 U, 20-150 U, or 100-150 U per treatment. In yet a further example, a subject suffering from low back pain is administered about 15 U, about 20 U, about 25 U, about 30 U, about 35 U, about 40 U, about 45 U, about 50 U, about 55 U, about 60 U, about 65 U, about 70 U, about 75 U, about 80 U, about 85 U, about 90 U, about 95 U, about 100 U, about 105 U, about 110 U, about 115 U, about 120 U, about 125 U, about 130 U, about 135 U, about 140 U, about 145 U, or about 150 U per treatment. Doses greater than 150 U per treatment can also be administered to a patient suffering from low back pain to achieve a therapeutic response. A treatment session can include multiple treatments.

[0212] In some embodiments, the patient suffers from migraines, including those suffering from migraines for at least 4 hours and at least 15 days per month. For example, a subject suffering from migraines is administered about 0.5-200 U of the pharmaceutical formulation of the present invention per treatment. In further examples, a subject suffering from migraines is administered about 5-190 U, 15-180 U, 25-170 U, 35-160 U, 45-150 U, 55-140 U, 65-130 U, 75-120 U, 85-110 U, or 95-105 U per treatment. A treatment session may include multiple treatments.

[0213] For example, about 0.5 U, about 1.0 U, about 1.5 U, about 2.0 U, about 2.5 U, about 3.0 U, about 3.5 U, about 4.0 U, about 4.5 U, about 5.0 U, about 5.5 U, about 6.0 U, about 6.5 U, about 7.0 U, about 7.5 U, about 8.0 U, about 8.5 U, about 9.0 U, about 9.5 U, about 10.0 U, about 12 U, about 15 U, about 17 U, about 20 U, about 22 U, about 25 U, about 27 U, about 30 U, about 32 U, about 35 U, about 37 U, about 40 U, about 42 U, about 45 U, about 47 U, or about 50 U per treatment is administered to a migraine patient. A patient can be treated at multiple sites, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or more. In one embodiment, a patient suffering from migraine headaches receives 31 injections using 5 U per 0.1 mL injection: corrugator supercilii (2 injections of 5 U each), procerus (1 injection of 5 U), frontalis (4 injections of 5 U each), temporalis (8 injections of 5 U each), occipitalis (6 injections of 5 U each), cervical paraspinal muscles (4 injections of 5 U each), and trapezius (6 injections of 5 U each). With the exception of the procerus, which can be injected in the midline, in certain embodiments, all muscles can be injected bilaterally, with half of the injection sites on the left side of the head and neck and half on the right side. Doses of more than 200 U per treatment can also be administered to migraine headache patients to achieve a therapeutic response. A treatment session can include multiple treatments.

[0214] In one embodiment, the patient is suffering from a sinus headache. For example, a subject suffering from a sinus headache is administered about 4-40 U per treatment of a pharmaceutical formulation of the present invention. In a further example, a subject suffering from a sinus headache is administered about 4-10 U, 4-15 U, 4-20 U, 4-25 U, 4-30 U, 4-35 U, 7-15 U, 7-20 U, 7-25 U, 7-30 U, 7-35 U, or 7-40 U per treatment. In yet a further example, a subject suffering from sinus headaches is administered about 4 U, about 5 U, about 7.5 U, about 10 U, about 12.5 U, about 15 U, about 17.5 U, about 20.0 U, about 22.5 U, about 25.0 U, about 27.5 U, about 30.0 U, about 32.5 U, about 35 U, about 37.5 U, or about 40 U per treatment. Doses greater than 40 U per treatment can also be administered to a sinus headache patient to achieve a therapeutic response. A treatment session can include multiple treatments.

[0215] In one embodiment, the patient is suffering from tension headache. For example, a subject suffering from tension headache is administered about 5-50 U of the pharmaceutical formulation of the present invention per treatment. In a further example, a subject is administered about 5-10 U, 5-15 U, 5-20 U, 5-25 U, 5-30 U, 5-35 U, 5-40 U, 5-45 U, 10-20 U, 10-25 U, 10-30 U, 10-35 U, 10-40 U, or 10-45 U per treatment. In yet a further example, a subject is administered about 5 U, 10 U, 20 U, 25 U, 30 U, 35 U, 40 U, 45 U, or 50 U per treatment. In one embodiment, a patient suffering from tension headaches is injected 31 times using 5 U per 0.1 mL injection: corrugator supercilii (2 injections of 5 U each), procerus (1 injection of 5 U), frontalis (4 injections of 5 U each), temporalis (8 injections of 5 U each), occipitalis (6 injections of 5 U each), cervical paraspinal muscles (4 injections of 5 U each), and trapezius (6 injections of 5 U each). With the exception of the procerus, which can be injected in the midline, in certain embodiments, all muscles can be injected bilaterally, with half of the injection sites on the left side of the head and neck and half on the right side. Doses of more than 200 U per treatment can also be administered to tension headache patients to achieve a therapeutic response. A treatment session can include multiple treatments.

[0216] In some embodiments, the patient suffers from sinus headache or facial pain associated with acute or recurrent chronic sinusitis. For example, a pharmaceutical formulation of the present invention can be administered to the nasal mucosa or subcutaneous structures covering the sinuses, and administration of the formulation reduces headache and / or facial pain associated with acute recurrent or chronic sinusitis. In further embodiments, any of the pharmaceutical formulations of the present invention can be administered to the nasal mucosa or subcutaneous structures covering the upper part of one or more sinuses, such as the ethmoid sinus, maxillary sinus, mammary gland, frontal sinus, and sphenoid sinus. In another embodiment, the subcutaneous structures covering the sinuses are located in one or more areas selected from the group consisting of the forehead, cheek, temporal region, postauricular region, and lip region. In some embodiments, multiple injections of 5 U each are administered to treat sinus headache or facial pain associated with acute or recurrent chronic sinusitis.

[0217] In another embodiment, a patient suffering from sinus headache or facial pain associated with acute or recurrent chronic sinusitis is treated by administering any of the pharmaceutical formulations of the present invention to the affected area of the patient. In a further embodiment, the pharmaceutical formulations disclosed herein are administered to the processes of the trigeminal nerve, which innervates the paranasal sinuses.

[0218] Patients suffering from sinus headaches or facial pain associated with acute or recurrent chronic sinusitis often present with symptoms including rhinitis, excessive sinus secretions, and / or purulent nasal discharge. In one embodiment, patients treated with the pharmaceutical formulations of the present invention present with symptoms of excessive sinus secretions and purulent nasal discharge.

[0219]

[0010] Embodiments of the present invention also provide methods for treating a patient suffering from sinus headache or facial pain associated with acute or recurrent chronic sinusitis, wherein the patient suffers from neuralgia. In certain embodiments, the neuralgia is trigeminal neuralgia. In other embodiments, the neuralgia is associated with compressive forces on a sensory nerve, with intrinsic nerve injury, with a demyelinating disease, or with a genetic disease, with a metabolic disease, with a central vascular disease, or with trauma. In other embodiments of the present invention, the pain is associated with tooth extraction or dental reconstruction. Treatment of urinary system disorders

[0220] In some embodiments, the present invention also provides a method for treating a patient suffering from overactive bladder (OAB), such as that caused by a neurological condition (NOAB) or idiopathic OAB (IOAB). For example, the pharmaceutical formulations of the present invention can be administered to or near the bladder, such as the detrusor muscle, and administration of the formulation reduces urge incontinence associated with overactive bladder. In certain embodiments, the dosage can be, for example, 200 U, or more, or less. For example, dosages can be about 15 U, about 20 U, about 25 U, about 30 U, about 35 U, about 40 U, about 45 U, about 50 U, about 55 U, about 60 U, about 65 U, about 70 U, about 75 U, about 80 U, about 85 U, about 90 U, about 95 U, about 100 U, about 105 U, about 110 U, about 115 U, about 120 U, about 125 U, about 130 U, about 135 U, about 140 U, about 145 U, about 150 U, about 160 U, about 170 U, about 180 U, about 190 U, about 200 U, about 210 U, about 220, about 230 U, about 240 U, or more per treatment. A patient can be injected at multiple sites, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, or more. In one embodiment, a patient suffering from OAB is treated by injecting 301 mL of approximately 6.7 U per injection into the detrusor muscle.

[0221] In some embodiments, the present invention also provides a method for treating a patient suffering from neurogenic detrusor overactivity (NDO), such as that caused by a neurological condition. For example, the pharmaceutical formulation of the present invention can be administered to or near the bladder, e.g., the detrusor, and administration of the formulation reduces urge urinary incontinence associated with overactive bladder. In certain embodiments, the dosage can be, for example, 200 U, or more, or less. For example, dosages can be about 15 U, about 20 U, about 25 U, about 30 U, about 35 U, about 40 U, about 45 U, about 50 U, about 55 U, about 60 U, about 65 U, about 70 U, about 75 U, about 80 U, about 85 U, about 90 U, about 95 U, about 100 U, about 105 U, about 110 U, about 115 U, about 120 U, about 125 U, about 130 U, about 135 U, about 140 U, about 145 U, about 150 U, about 160 U, about 170 U, about 180 U, about 190 U, about 200 U, about 210 U, about 220, about 230 U, about 240 U, or more per treatment. A patient can be infused at multiple sites, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or more. In one embodiment, a patient suffering from NDO is treated by injecting 301 mL of approximately 6.7 U per injection into the detrusor muscle. Treatment of cosmetic features

[0222] In another embodiment, the present invention provides a method comprising administering to a subject in need thereof at least one pharmaceutical formulation of the present invention in an amount sufficient to restore a cosmetically altered soft tissue feature, hi a further embodiment, the pharmaceutical formulation is administered by transdermal or transmucosal injection at either a single site or multiple sites.

[0223] In an embodiment, the pharmaceutical formulation of the present invention is administered to the face or neck of a subject. In a further embodiment, the pharmaceutical formulation of the present invention is administered to a subject in an amount sufficient to reduce wrinkles. For example, the formulation can be administered to the subject's glabella in an amount sufficient to reduce vertical wrinkles between the glabella and the bridge of the nose. The pharmaceutical formulation can also be administered near one or both of the subject's eyes in an amount sufficient to reduce wrinkles at the corners of the eyes. In one embodiment, the composition of the present invention can be topically injected into smooth skin. In another embodiment, the pharmaceutical formulation of the present invention can also be administered to the subject's forehead in an amount sufficient to reduce horizontal wrinkles on the forehead. In yet another embodiment of the present invention, the pharmaceutical formulation is administered to the subject's neck in an amount sufficient to reduce muscle stiffness in the neck. In one embodiment, the pharmaceutical composition is applied to the masseter muscle for muscle relaxation and / or reduction of masseter muscle mass.

[0224] In a further embodiment, the patient is suffering from facial wrinkles. A subject suffering from facial wrinkles can be administered, for example, about 1 to 100 U of the pharmaceutical formulation of the present invention per treatment. In a further example, a subject is administered about 1 to 10 U, 1 to 20 U, 1 to 30 U, 1 to 40 U, 1 to 50 U, 1 to 60 U, 1 to 70 U, 1 to 80 U, 1 to 90 U, 5 to 20 U, 5 to 30 U, 5 to 40 U, 5 to 50 U, 5 to 60 U, 5 to 70 U, 5 to 80 U, 5 to 90 U, or 5 to 100 U per treatment for an inflammatory disorder. In yet a further example, a patient is administered about 1 U, about 10 U, about 20 U, about 30 U, about 40 U, about 50 U, about 60 U, about 70 U, about 80 U, about 90 U, or about 100 U per treatment. Doses of greater than 100 U per treatment may be administered to patients suffering from inflammation or an inflammatory disorder to achieve a therapeutic response. Treating inflammation

[0225] In another embodiment, the present invention provides a method comprising administering a pharmaceutical formulation of the present invention to a subject in need of treatment for inflammation in an amount sufficient to reduce inflammation. In certain embodiments, the pharmaceutical formulation of the present invention is administered to a patient without causing muscle weakness. In some embodiments, the pharmaceutical formulation of the present invention is administered to a patient with an inflammatory condition. In certain embodiments, the inflammatory condition is neurogenic inflammation. In another embodiment, the subject suffers from rheumatoid arthritis or a gastrointestinal inflammatory disease.

[0226] In a further embodiment, the patient suffers from an inflammatory disorder. A subject suffering from an inflammatory disorder is administered, for example, about 1 to 100 U per treatment of a pharmaceutical formulation of the present invention. In a further example, a subject is administered about 1 to 10 U, 1 to 20 U, 1 to 30 U, 1 to 40 U, 1 to 50 U, 1 to 60 U, 1 to 70 U, 1 to 80 U, 1 to 90 U, 5 to 20 U, 5 to 30 U, 5 to 40 U, 5 to 50 U, 5 to 60 U, 5 to 70 U, 5 to 80 U, 5 to 90 U, or 5 to 100 U per treatment. In yet a further example, a patient is administered about 1 U, about 10 U, about 20 U, about 30 U, about 40 U, about 50 U, about 60 U, about 70 U, about 80 U, about 90 U, or about 100 U per treatment. Doses of greater than 100 U per treatment may be administered to patients suffering from inflammation or an inflammatory disorder to achieve a therapeutic response. Treatment of skin symptoms

[0227] Methods within the scope of the present invention for treating skin disorders can include the step of topically administering a botulinum neurotoxin to the location of the skin disorder, such as the patient's face, hands, or feet. The neurotoxin may be topically administered in an amount of about 10-3 units / kg to about 35 units / kg of patient body weight. For example, the neurotoxin may be topically administered in an amount of about 10-2 U / kg to about 25 U / kg of patient body weight. In a further example, the neurotoxin is administered in an amount of about 10-1 U / kg to about 15 U / kg. In one method within the scope of the present invention, the neurotoxin is topically administered in an amount of about 1 U / kg to about 10 U / kg. In a clinical setting, it may be advantageous to administer 1 U to 3000 U of a neurotoxin, such as botulinum toxin type A or B, to the location of the skin disorder via transdermal or subcutaneous application to effectively treat the skin disorder.

[0228] Botulinum toxin may be administered at multiple sites on the skin, with adjacent injection sites spaced about 0.1 to 10 cm apart, or about 0.5 to about 5 cm apart, e.g., about 1.5 to about 3 cm apart. The toxin may be any of botulinum toxin types A, B, C, D, E, F, or G. The amount administered may vary from 0.1 to 1000 U, or from about 1 to about 40 U, or from about 5 to about 10 U, depending on the manufacturer's specifications, the type of toxin, and the method of administration. The repetition period for these administrations to maintain the desired changes varies substantially depending on the site of injection, the symptoms being controlled, and the patient's condition. Thus, the repetition period may vary from about 1 week to about 50 weeks, with a typical range being from about 4 to about 25 weeks, or even about 12 to about 16 weeks.

[0229] The distance between injections, e.g., the distance between injections, can vary from about 1 mm to about 10 cm, preferably from about 5 mm to about 5 cm, and more usually from about 1 cm to about 3 cm. Thus, for example, botulinum type A can be suitably administered by intradermal injection at about 0.1 to about 10 U, spaced about 0.5 to about 10 cm apart.

[0230] In another embodiment, the present invention provides a method comprising administering to a subject in need of treatment for a skin disorder a pharmaceutical formulation of the present invention in an amount sufficient to reduce sebum or mucus secretion. In a further embodiment, the pharmaceutical formulation of the present invention is administered to a patient without causing muscle weakness. In certain embodiments, the pharmaceutical formulation of the present invention is injected into one or more sites of the eyelid or conjunctiva. In another embodiment, the formulation of the present invention is administered to a body surface.

[0231] In another embodiment, the pharmaceutical formulation is administered in an amount sufficient to reduce bacterial or fungal growth on the skin, including, but not limited to, Staphylococcus, Streptococcus, and Moraxella. For example, the pharmaceutical formulation of the present invention is administered to an area selected from the group consisting of the eyelids, scalp, feet, groin, and armpits to reduce skin infections. [Example]

[0232] The following examples illustrate embodiments and aspects of the present invention and are not intended to limit the scope of the invention. Example 1 Activity and stability of representative solid clostridial pharmaceutical compositions relative to prior art formulations.

[0233] Bulk solutions of botulinum toxin were prepared by mixing appropriate amounts of botulinum toxin type A with several vehicle solutions listed in Tables 1-3 below. The solutions were filled into glass vials and lyophilized using conventional lyophilization conditions. After reconstitution of the lyophilizates with saline, the potency of the lyophilized formulations was tested by cell-based potency assay (CBPA). Potency recovery results after lyophilization and storage at the indicated temperatures are tabulated and normalized to target potency. The potency of solid compositions prepared according to embodiments of the present invention was compared to three comparative formulations listed in Tables 1-3. [Table 25] Treh = trehalose, P188 = poloxamer P188, Met = L-methionine, NAC = N-acetyl-L-cysteine. [Table 26] Treh = trehalose, P188 = poloxamer P188, Met = L-methionine, NAC = N-acetyl-L-cysteine. [Table 27] Treh = trehalose, Sucr = sucrose, P188 = poloxamer P188, Met = L-methionine, NAC = N-acetyl-L-cysteine. [Table 28] P-188 = Poloxamer P-188, Met = L-methionine. Example 2 Activity of Liquid Clostridial Pharmaceutical Compositions in the Presence or Absence of Antioxidants

[0234] Bulk drug product solutions were prepared by mixing appropriate amounts of botulinum toxin type A with three different vehicle solutions, as shown in Table 4. All three formulations contained 8% w / w trehalose, 4% w / w P188, and 20 mM histidine buffer, pH 6.0. The target potency was 100 units / mL. Formulation 10 contained no antioxidants. Formulations 11 and 12 contained NAC and methionine, respectively. The bulk solutions were filled into 2 mL glass vials (fill volume 1.25 mL) and sealed with rubber stoppers and aluminum shells. The potencies of the formulations were tested by cell-based potency assay (CBPA) after filling (time zero, t0) and storage at four temperatures (-70, 5, 25, and 40°C) for one month. The potency test results are shown in Table 4. Briefly, the methionine-containing formulation (#12) retained its potency after 1 month of storage at all four temperatures, including 40°C, whereas the antioxidant-free formulation lost approximately 17% of its potency at 25°C and essentially all activity at 40°C (i.e., complete inactivation). The second antioxidant tested, N-acetyl-L-cysteine, lost potency more rapidly compared to the antioxidant-free formulation, indicating that N-acetyl-L-cysteine acted as a pro-oxidant in this formulation. The liquid composition containing NAC lost 50% of its potency after 1 month of storage at 25°C and essentially all activity after 1 month of storage at 40°C (i.e., complete inactivation of the toxin). In contrast, as shown in Table 1, the lyophilized composition containing NAC lost 7% potency ((98.6%-91.9%) / 98.6%) after 3 months of storage at 25°C and 12% potency ((98.6%-86.5%) / 98.6%) after 1 month of storage at 40°C, indicating that NAC can function as a stabilizer in the lyophilized composition. [Table 29] * Each formulation contained the same amount of botulinum toxin, 8 w / w% trehalose, and 4 w / w% poloxamer P188 in a histidine buffer. Example 3 Effect of representative antioxidants on the stability of representative liquid formulations.

[0235] Bulk drug product solutions were prepared by mixing appropriate amounts of botulinum toxin type A with different antioxidants as shown in Table 5. All formulations contained 8% w / w trehalose, 4% w / w P188, and 20 mM histidine buffer, pH 6.0, and one or more antioxidants as indicated. The target potency was 100 U / mL. The bulk solutions were filled into 2 mL glass vials (fill volume 1.25 mL) and sealed with rubber stoppers and aluminum shells. The potencies of the formulations were tested by cell-based potency assay (CBPA) after filling (time zero, t0) and after storage at 40°C for 2 weeks and 1 month. The potency test results are shown in Table 5. The liquid composition containing NAC and EDTA sodium salt (Formulation 26) retained full potency after storage at 40°C for 2 weeks and 1 month, as did the composition containing NAC, EDTA sodium salt, and tryptophan (Formulation 27). In contrast, as shown in Table 5, a liquid composition containing NAC but no EDTA (Formulation 25) lost essentially all efficacy after 2 weeks of storage at 40° C. This indicates that the combination of antioxidant-chelating agents (e.g., EDTA, EGTA, or DTPA) and / or chain-terminating antioxidants (e.g., methionine, cysteine, NAC, and BHT) provides a stabilizing effect against botulinum toxin. [Table 30] 1 Each formulation contained 100 U / mL botulinum toxin, 8 w / w% trehalose, and 4 w / w% poloxamer P188 in 20 mM histidine buffer, pH 6.0, and specified antioxidants. 2 NAC = N-acetyl-L-cysteine, Met = L-methionine, TRP = L-tryptophan, GSH = L-glutathione, NaSul = sodium sulfite, PrpGal = propyl gallate, EDTA = ethylenediaminetetraacetic acid, sodium salt. 3 NT = Not Tested Example 4 The effect of surfactant selection on the stability of representative liquid compositions.

[0236] Bulk drug product solutions were prepared by mixing the appropriate amounts of botulinum toxin type A with the poloxamer or polysorbate indicated in Table 6. All formulations contained 8% w / w trehalose, 0.2% w / w methionine in 20 mM histidine buffer, pH 6.0, and 4% w / w P188 or 0.04% w / w Tween-20 as indicated. The target potency was 100 U / mL. The bulk solution was filled into 2 mL glass vials (1.25 mL fill volume) and sealed with rubber stoppers and aluminum shells. After filling to obtain the initial potency, the potency of the formulations was tested by cell-based potency assay (CBPA). After storage at 40°C for 1 month, potency was measured again. The potency test results are shown in Table 6. As can be seen from the results in Table 6, the liquid formulations containing poloxamer provided improved stability after 1 month of storage at 40°C compared to the corresponding formulations containing polysorbate. [Table 31] Tre = trehalose, P-188 = poloxamer-188, Tw-20 = Tween-20, Met = L-methionine; His = L-histidine. CBPA provides residual activity in U / mL. Example 5 Effect of omitting a tonicity agent on the stability of a representative liquid composition.

[0237] Bulk drug product solutions were prepared by mixing appropriate amounts of botulinum toxin type A with different ingredients as shown in Tables 7 and 8. The amounts of the ingredients included were 0% or 8% w / w trehalose, 0% or 0.2% w / w methionine, and 0% or 4% w / w P188, all formulated in 20 mM histidine buffer (pH 6.0). The target potencies were 100, 150, or 200 U / mL. The bulk solutions were filled into 2 mL glass vials (fill volume 1.25 mL) and sealed with rubber stoppers and aluminum shells. After filling to obtain the initial potency, the potency of the formulations was tested by cell-based potency assay (CBPA). After storage at -70°C and 40°C for 1 month, potency was measured again. The potency test results are shown in Tables 7 and 8. As can be seen from the results in these tables, the liquid formulation containing poloxamer and no tonicity agent provides excellent stability after one month of storage at -70°C and 40°C. This is supported by the results in Table 10 below for Formulation 40. On the other hand, the liquid formulation containing polysorbate and no tonicity agent provides some stability after one month of storage at -70°C. [Table 32] [Table 33] Sucr = sucrose, Tre = trehalose, P-188 = poloxamer-188, Tw-20 = Tween-20, Met = L-methionine, His = L-histidine. Initial = initial activity in U / mL Results after 1 month are residual activity in U / mL. Example 6 Stability of various liquid compositions

[0238] Bulk drug product solutions were prepared by mixing appropriate amounts of botulinum toxin type A with vehicle solutions containing different stabilizers and surfactants, as shown in Table 9. All formulations contained 8% or 0% w / w trehalose or sucrose, 4% w / w P188 or 0.04% w / w PS-20, optionally 20 mM histidine buffer (pH 6.0), 0.2% w / w methionine, and optionally 0.9% w / w NaCl. The target potency was 100 U / mL. The bulk solution was filled into 2 mL glass vials (fill volume 1.25 mL) and sealed with rubber stoppers and aluminum shells. The potencies of the formulations were tested by cell-based potency assay (CBPA) after filling (time zero, t0) and after storage at 5°C. The potency test results are shown in Table 10. [Table 34] Suc = sucrose, Tre = trehalose, P-188 = poloxamer-188, Ps-20 = Tween-20, Met = L-methionine, His = L-histidine. [Table 35] The result is residual activity in U / mL. Example 7 Effect of sugar choice on the stability of representative lyophilized formulations.

[0239] Bulk solutions of botulinum toxin were prepared by mixing appropriate amounts of botulinum toxin type A with several vehicle solutions listed in Table 11. The amounts of the components included 0% or 8% w / w trehalose or sucrose, 0.2% w / w methionine, 0% or 4% w / w P188, and 0% or 0.04% w / w Tween 20, all formulated in 20 mM histidine buffer (pH 6.0). The target potency was 200 U / mL. These amounts are shown in Table 11 below. The solutions were filled into glass vials and lyophilized using conventional lyophilization conditions. After reconstitution of the lyophilized compositions after 2 weeks of storage at -70°C or 40°C, the potency of the lyophilized formulations was tested by cell-based potency assay (CBPA). Potency recovery results are shown in Table 12. The effect of sugar type on these lyophilized formulations can be deduced by comparing Formulation No. 42 with Formulation No. 44 and Formulation No. 43 with Formulation No. 45. The potency of botulinum toxin in Formulations 42 and 44 after 2 weeks of storage at -20°C and 40°C reveals that higher potency recovery was achieved in the lyophilized compositions containing trehalose compared to the lyophilized compositions containing sucrose. [Table 36] Tre = trehalose, Sucr = sucrose, Tw-20 = Tween-20, P-188 = poloxamer-188, Met = L-methionine, His = L-histidine. [Table 37] Example 8 Effect of surfactant choice on the stability of representative lyophilized compositions.

[0240] The results in Tables 11 and 12 also demonstrate the effect of surfactant choice on the stability of particular lyophilized compositions. By comparing Formulation No. 42 with Formulation No. 43 and Formulation No. 44 with Formulation No. 45, the effect of surfactant type on these lyophilized formulations can be deduced. The potency of botulinum toxin in Formulations 43 and 45 after two weeks of storage at -20°C and 40°C reveals that higher potency recovery was achieved in lyophilized compositions containing a poloxamer surfactant compared to lyophilized compositions containing a polysorbate surfactant. Example 9 Treatment of Depression with Representative Compositions

[0241] A 58-year-old university professor presented with symptoms of depression. A physician diagnosed the patient with moderate-to-severe major depressive disorder (MDD) based on DSM-IV-TR criteria. The physician administered a botulinum toxin solution prepared using the composition shown in Table A, reconstituted with saline, via intramuscular injection into the procerus and corrugator supercilii muscles in a single treatment session. A total of 30 units was administered over six injection sites. The procerus and corrugator supercilii muscles were numbed. Treatment was subsequently confirmed to have alleviated depression using the Montgomery Asberg Depression Rating Scale. Additional efficacy measures, such as a clinical CGI-S score (Clinical Global Impression of Change score) and a clinical HAM-D17 total score (Hamilton Rating Scale for Depression), further confirmed the treatment. After three months, treatment was repeated. Example 10 Treatment of Depression with Representative Compositions

[0242] A 21-year-old student presents with symptoms of depression. A physician diagnoses one symptom of moderate-to-severe major depressive disorder (MDD) based on DSM-IV-TR criteria. The physician administers a botulinum toxin solution prepared using the composition shown in Table A, reconstituted with saline, via intramuscular injection into the procerus and corrugator supercilii muscles in a single treatment session. A total of 50 units are administered over eight injection sites. The procerus and corrugator supercilii muscles are paralyzed. Treatment is subsequently confirmed to have alleviated depression using the Montgomery Asberg Depression Rating Scale. Additional efficacy measures, such as a clinical CGI-S score (Clinical Global Impression of Change score) and a clinical HAM-D17 total score (Hamilton Rating Scale for Depression), further confirm the treatment. After six months, treatment is repeated. Example 11 Treatment of Cardiac Arrhythmias with Representative Compositions

[0243] A 41-year-old teacher undergoing cardiac surgery presents with symptoms of cardiac arrhythmia. The surgeon administers a botulinum toxin solution made using the composition shown in Table A, reconstituted with saline, into the patient's major epicardial fat pads. 50 U are administered per epicardial fat pad, up to a total of 250 U. The patient is followed by ECG for four weeks after administration. The incidence of atrial fibrillation is reduced. Additionally, the following improvements are noted for this patient: shorter hospital stay, shorter ICU stay, reduced anticoagulant use, and a reduced need for postoperative atrial fibrillation (POAF) interventions, such as ablation, pacemaker implantation, or electrical or pharmacological cardioversion.

[0244] Example 12: Treatment of cardiac arrhythmias with representative compositions.

[0245] A 66-year-old pensioner undergoing cardiac surgery presents with symptoms of cardiac arrhythmia. The surgeon administers a botulinum toxin solution made using the composition shown in Table A, reconstituted with saline, into the patient's main epicardial fat pad. 25 U are administered per epicardial fat pad, up to a total of 125 U. The patient is followed by ECG for 4 weeks after administration. The incidence of atrial fibrillation is reduced. Additionally, the following improvements are noted for this patient: shorter hospital stay, shorter ICU stay, reduced anticoagulant use, and a reduced need for interventional procedures for postoperative atrial fibrillation (POAF), such as ablation, pacemaker implantation, or electrical or pharmacological cardioversion.

[0246] Numerous changes and modifications may be made by those skilled in the art without departing from the spirit and scope of the present disclosure. Accordingly, it should be understood that the described embodiments are presented for illustrative purposes only, and that the embodiments should not be considered as limiting the scope of the following claims. Accordingly, the following claims should be read to include not only the literally recited combinations of elements, but all equivalent elements that perform substantially the same function in substantially the same way to achieve substantially the same result. In this manner, the claims should be understood to include what has been described above, what is conceptually equivalent, and what encompasses the concepts of the present disclosure.

[0247] The present disclosure also includes the following series of items: 1. A pharmaceutical composition comprising: (i) a clostridial toxin active ingredient; and (ii) a tonicity agent and / or a lyoprotectant; and (iii) poloxamer and / or polysorbate, (iv) an antioxidant. 2. The composition according to item 1, comprising a botulinum toxin. 3. The composition according to item 1 or 2, comprising trehalose. 4. The composition according to any one of items 1 to 3, comprising poloxamer 188 and / or polysorbate 20. 5. The composition according to any one of items 1 to 4, comprising one or more of methionine and N-acetyl-cysteine. 6. The composition according to any one of items 1 to 5, comprising a botulinum toxin, trehalose, one of poloxamer 188 or polysorbate 20, and one of methionine or N-acetyl-cysteine. 7. The composition according to any one of items 1 to 6, comprising a botulinum toxin, trehalose, poloxamer 188, and methionine. 8. The composition according to item 7, wherein the relative weight amounts (%, w / w) of trehalose, poloxamer 188, and methionine are within the following ranges: [Table 38] 9. The composition according to any one of items 1 to 6, comprising a botulinum toxin, trehalose, polysorbate 20, and methionine. 10. The composition according to item 9, wherein the relative weight amounts (%, w / w) of trehalose, polysorbate 20, and methionine are within the following ranges: [Table 39] 11. The composition according to any one of items 1 to 6, comprising a botulinum toxin, trehalose, poloxamer 188, and N-acetyl-cysteine. 12. The composition according to item 11, wherein the relative weight amounts (%, w / w) of trehalose, poloxamer 188, and N-acetyl-cysteine are within the following ranges: [Table 40] 13. The composition according to any one of items 1 to 6, comprising a botulinum toxin, trehalose, polysorbate 20, and N-acetyl-cysteine. 14. The composition according to 13, wherein the relative weight amounts (%, w / w) of trehalose, polysorbate 20, and N-acetyl-cysteine are within the following ranges: [Table 41] 15. The composition according to any one of items 1 to 14, comprising histidine. 16. The composition according to any one of items 1 to 15, which does not contain animal-derived proteins. 17. The composition according to any one of items 1 to 4, comprising ethylenediaminetetraacetic acid sodium salt (EDTA) or an EDTA analogue. 18. The composition according to any one of items 5 and 6, further comprising ethylenediaminetetraacetic acid sodium salt (EDTA) or an EDTA analogue. 19. The composition according to any one of items 11 to 14, further comprising ethylenediaminetetraacetic acid sodium salt (EDTA) or an EDTA analogue. 20. The composition according to any one of items 1 to 19, wherein the relative weight (%, w / w) of EDTA is in the range of about 0.01 to 0.10. 21. The composition according to any one of items 1 to 20, wherein the composition is a solid formulation. 22. The composition according to any one of items 12, 14, 19 and 20, wherein the composition is a solid formulation and the relative weight (%, w / w) of N-acetyl-cysteine is 0.01 to 0.05. 23. The composition according to any one of items 1 to 20, wherein the composition is a liquid formulation and has a pH of 5 to 7. 24. The composition according to any one of items 11 to 14, 19 and 20, wherein the composition is a liquid formulation, has a pH of 5 to 7 and a relative weight amount of N-acetyl-cysteine of 0.1 to 0.5.

[0248] The present disclosure also includes the following set of embodiments. 1. A pharmaceutical composition comprising: (i) a clostridial toxin active ingredient; and (ii) a tonicity agent and / or a lyoprotectant; and (iii) poloxamer and / or polysorbate, (iv) an antioxidant. 2. The composition of embodiment 1, comprising a botulinum toxin. 3. The composition of embodiment 1 or embodiment 2, comprising trehalose or sucrose. 4. The composition of any one of embodiments 1 to 3, comprising poloxamer 188 and / or polysorbate 20. 5. The composition of any one of embodiments 1 to 4, comprising one or more of methionine and N-acetyl-cysteine. 6. The composition of any one of embodiments 1-5, comprising a botulinum toxin, trehalose or sucrose, one of poloxamer 188 or polysorbate 20, and one of methionine or N-acetyl-cysteine. 7. The composition of any one of embodiments 1 to 6, comprising a botulinum toxin, trehalose or sucrose, poloxamer 188, and methionine. 8. The composition of embodiment 7, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, and methionine are within the following ranges: [Table 42] 9. The composition of embodiment 8, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, and methionine are as follows: [Table 43] 10. The composition according to any one of embodiments 7 to 9, wherein the composition is a liquid composition, and the composition preferably comprises a buffer. 11. The composition of embodiment 10, wherein the buffering agent is histidine, the concentration of histidine is preferably 20 mM, the pH is preferably in the range of 5 to 7, the pH is more preferably in the range of 5.5 to 6.5, and the pH is most preferably in the range of 5.5 to 6. 12. The composition of embodiment 10, wherein the composition consists of trehalose or sucrose, poloxamer 188, methionine, a buffering agent, and optionally NaCl, and optionally EDTA, wherein the buffering agent is preferably histidine, and wherein the pH is preferably in the range of 5 to 7, more preferably in the range of 5.5 to 6.5, and most preferably in the range of 5.5 to 6. 13. The composition according to any one of embodiments 7 to 9, wherein the composition is a solid composition, and the composition is preferably lyophilized. 14. The composition according to embodiment 13, further comprising a buffering agent, preferably histidine, and preferably containing histidine in a relative weight amount of 0.1 to 0.5 (% w / w), more preferably 0.3 to 0.4% (w / w). 15. The composition of embodiment 13 or 14, wherein the composition consists of trehalose or sucrose, poloxamer 188, methionine, a buffer, and optionally NaCl, and optionally EDTA. 16. The composition of any one of embodiments 1-6, comprising a botulinum toxin, trehalose or sucrose, polysorbate 20, and methionine. 17. The composition of embodiment 16, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, polysorbate 20, and methionine are within the following ranges: [Table 44] 18. The composition of any one of embodiments 1 to 6, comprising a botulinum toxin, trehalose or sucrose, poloxamer 188, and N-acetyl-cysteine. 19. The composition of embodiment 18, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, and N-acetyl-cysteine are within the following ranges: [Table 45] 20. The composition of embodiment 19, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, N-acetyl-cysteine, and EDTA are as follows: [Table 46] 21. The composition of embodiment 19, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, and N-acetyl-cysteine are as follows: [Table 47] 22. The composition according to any one of embodiments 18 to 21, wherein the composition is a solid composition, and the composition is preferably lyophilized. 23. The composition according to embodiment 22, further comprising a buffering agent, preferably histidine, and preferably containing histidine in a relative weight amount of 0.1 to 0.5 (% w / w), more preferably 0.3 to 0.4% (w / w). 24. The composition of embodiment 22 or 23, wherein the composition consists of trehalose or sucrose, poloxamer 188, N-acetyl-cysteine, and a buffering agent. 25. The composition according to any one of embodiments 18 to 20, wherein the composition is a liquid composition, and the composition preferably comprises a buffer. 26. The composition of embodiment 25, wherein the buffering agent is histidine, the concentration of histidine is preferably 20 mM, and the pH is preferably in the range of 5 to 7, with the pH being most preferably 6. 27. The composition of embodiment 25, wherein the composition consists of trehalose or sucrose, poloxamer 188, N-acetyl-cysteine, a buffer, and optionally EDTA, wherein the buffer is preferably histidine, and wherein the pH is preferably within the range of 5 to 7, and most preferably 6. 28. The composition of any one of embodiments 1-6, comprising a botulinum toxin, trehalose or sucrose, polysorbate 20, and N-acetyl-cysteine. 29. The composition of embodiment 28, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, polysorbate 20, and N-acetyl-cysteine are within the following ranges: [Table 48] 30. A pharmaceutical composition comprising: (i) a clostridial toxin active ingredient; and (ii) poloxamer and / or polysorbate, (iii) an antioxidant; A pharmaceutical composition, wherein no sugars or polyalcohols are present in the composition. 31. The composition of embodiment 30, comprising a botulinum toxin. 32. The composition of embodiment 30 or 31, comprising poloxamer 188 and / or polysorbate 20. 33. The composition of any one of embodiments 30-32, comprising one or more of methionine and N-acetyl-cysteine. 34. The composition of any one of embodiments 30-33, comprising a botulinum toxin, one of poloxamer 188 or polysorbate 20, and one of methionine or N-acetyl-cysteine. 35. The composition of any one of embodiments 30-34, comprising a botulinum toxin, poloxamer 188, and methionine. 36. The composition of embodiment 35, wherein the relative weight amounts (%, w / w) of poloxamer 188 and methionine are within the following ranges: [Table 49] 37. The composition of embodiment 36, wherein the relative weight amounts (%, w / w) of poloxamer 188 and methionine are as follows: [Table 50] 38. The composition according to any one of embodiments 35 to 37, wherein the composition is a liquid composition, and the composition preferably comprises a buffer. 39. The composition of embodiment 38, wherein the buffering agent is histidine, the concentration of histidine is preferably 20 mM, and the pH is preferably in the range of 5 to 7, with the pH being most preferably 6. 40. The composition described in embodiment 38, wherein the composition consists of poloxamer 188, methionine, and a buffering agent, wherein the buffering agent is preferably histidine, and wherein the pH is preferably within the range of 5 to 7, and wherein the pH is most preferably 6. 41. The composition of any one of embodiments 1-10, 12-25, 27-38, or 40, comprising histidine. 42. The composition of any one of claims 1 to 41, which is free of animal-derived proteins. 43. The composition of any one of claims 1 to 42, comprising ethylenediaminetetraacetic acid sodium salt (EDTA) or an EDTA analogue. 44. The composition of embodiment 43, wherein the relative weight (%, w / w) of EDTA is in the range of about 0.01 to 0.10. 45. The composition of any one of embodiments 1-9, 16-21, 28, 29, and 41-44, wherein the composition is a solid formulation. 46. The composition of any one of embodiments 19, 29, 43 or 44, wherein the composition is a solid formulation and the relative weight (%, w / w) of N-acetyl-cysteine is 0.01 to 0.05. 47. The composition of any one of embodiments 1-12, 16-21, and 25-44, wherein the composition is a liquid formulation and has a pH of 5-7. 48. The composition of any one of embodiments 18, 19, 28, 29, 43 and 44, wherein the composition is a liquid formulation, has a pH of 5 to 7, and the relative weight amount of N-acetyl-cysteine is 0.1 to 0.5. 49. A method for treating, reducing symptoms, and / or preventing diseases, disorders, and conditions, comprising administering to a subject in need thereof the pharmaceutical composition of any preceding embodiment. 50. The method of embodiment 49, wherein the disease, disorder, and condition is selected from neuromuscular diseases, pain, psychiatric disorders, urinary system disorders, inflammation, and skin diseases. 51. The method of embodiment 49, wherein the disorder is depression. 52. The method of embodiment 49, wherein the condition is cardiac arrhythmia. 53. A method of cosmetic treatment, comprising administering to a subject in need of cosmetic treatment a pharmaceutical composition according to any one of embodiments 1 to 48.

Claims

1. 1. A pharmaceutical composition comprising: (i) a botulinum toxin; (ii) poloxamer 188, and (iii) methionine or N-acetyl-cysteine.

2. 10. The composition of claim 1, further comprising trehalose or sucrose, optionally NaCl, and optionally EDTA.

3. 3. The composition of claim 2, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, methionine, and optionally NaCl, and optionally EDTA are within the following ranges: 【Table 1】

4. 4. The composition of claim 3, wherein the relative weight amounts (%, w / w) of trehalose or sucrose, poloxamer 188, methionine, and optionally NaCl, and optionally EDTA are as follows: 【Table 2】

5. 3. The composition of claim 2, wherein the relative weight percentages (w / w) of trehalose or sucrose, poloxamer 188, N-acetyl-cysteine, and EDTA are within the following ranges: 【Table 3】

6. 6. The composition of claim 5, wherein the relative weight percentages (w / w) of trehalose, poloxamer 188, N-acetyl-cysteine, and EDTA are as follows: 【Table 4】

7. The composition according to any one of claims 1 to 6, comprising trehalose.

8. The pharmaceutical composition of claim 1 , wherein the composition does not contain sugars or polyalcohols.

9. 9. The composition of claim 8, wherein the relative weights (%, w / w) of poloxamer 188 and methionine are within the following ranges: 【Table 5】

10. 10. The composition of claim 9, wherein the relative weights (%, w / w) of poloxamer 188 and methionine are as follows: 【Table 6】

11. The composition according to any one of claims 1 to 10, wherein the composition is a liquid composition, and the composition preferably comprises a buffer.

12. 12. The composition of claim 11, wherein the buffering agent is histidine, the concentration of histidine is preferably 20 mM, and the pH is preferably in the range of 5 to 7, most preferably in the range of 5.5 to 6.

13. The composition according to any one of claims 1 to 7, wherein the composition is a solid composition, and the composition is preferably freeze-dried.

14. The composition according to claim 13, further comprising a buffering agent, said buffering agent being preferably histidine, and the histidine is preferably contained in a relative weight amount of 0.1 to 0.5 (% w / w), more preferably 0.3 to 0.4% (w / w).

15. 5. The composition of any one of claims 1 to 4, wherein the composition is a liquid composition consisting of one or more botulinum toxins, trehalose or sucrose, poloxamer 188, methionine, a buffer, and optionally NaCl, and optionally EDTA.

16. 5. The composition of any one of claims 1 to 4, wherein the composition is a solid composition consisting of one or more botulinum toxins, trehalose, poloxamer 188, methionine, a buffer, and optionally NaCl, and optionally EDTA.

17. 7. The composition of any one of claims 1, 2, 5 and 6, wherein the composition is a liquid composition consisting of one or more botulinum toxins, trehalose or sucrose, poloxamer 188, N-acetyl-cysteine, a buffer, and EDTA.

18. The composition of any one of claims 1 and 8 to 10, wherein the composition is a liquid composition and consists of one or more botulinum toxins, poloxamer 188, methionine, and a buffering agent.

19. The composition according to any one of claims 1 to 18, which is free of animal-derived proteins.

20. 20. The composition of any one of claims 1 to 17 and 19, comprising ethylenediaminetetraacetic acid sodium salt (EDTA) or an EDTA analogue.

21. 21. The composition of claim 20, wherein the relative weight (%, w / w) of EDTA is in the range of about 0.01 to 0.

10.

22. 1. A liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a poloxamer; and (iii) a chelating agent; and (iv) a sacrificial antioxidant.

23. 23. The liquid pharmaceutical composition of claim 22, wherein the chelating agent is EDTA, EGTA, or DTP A and the sacrificial antioxidant is ascorbic acid.

24. 1. A liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a poloxamer; and (iii) a chelating agent; and (iv) a chain terminator.

25. 25. The liquid composition of claim 24, wherein the chelating agent is EDTA and the chain terminator is N-acetyl-cysteine.

26. 25. The liquid composition of claim 24, wherein the chelating agent is EDTA and the chain terminator is butylated hydroxytoluene.

27. 1. A liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a poloxamer; and (iii) methionine.

28. 1. A liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof; (iii) a poloxamer, and (iv) methionine.

29. 1. A liquid pharmaceutical composition comprising: (i) a clostridial toxin; (ii) a tonicity agent selected from trehalose, sucrose, sodium chloride, mannitol, sorbitol, glucose, and combinations thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a chain terminator.

30. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a sacrificial antioxidant.

31. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, and (iv) a chelating agent; and (v) a sacrificial antioxidant.

32. 32. The composition of any one of claims 30 to 31, wherein the chelating agent is EDTA, EGTA, or DTP A and the sacrificial antioxidant is ascorbic acid.

33. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, and (iv) a chelating agent; and (v) a chain terminator.

34. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chelating agent; and (v) a chain terminator.

35. The freeze-dried pharmaceutical composition according to any one of claims 33 to 34, wherein the chelating agent is EDTA and the chain terminator is N-acetyl-cysteine.

36. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, and (iv) a chain terminator.

37. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a surfactant selected from poloxamers, polysorbates, and combinations thereof; (iv) a chain terminator.

38. The freeze-dried pharmaceutical composition according to any one of claims 36 to 37, wherein the chain terminator is N-acetyl-cysteine.

39. 1. A lyophilized pharmaceutical composition comprising: (i) a clostridial toxin; (ii) trehalose, sucrose, mannitol, sorbitol, glucose, or a combination thereof; (iii) a poloxamer, and (iv) methionine.

40. 40. A method for treating, reducing symptoms, and / or preventing diseases, disorders, and conditions, comprising the step of administering to a subject in need thereof a pharmaceutical composition according to any one of claims 1 to 39.

41. 41. The method of claim 40, wherein the disease, disorder, or condition is selected from a neuromuscular disease, pain, a psychiatric disorder, a urinary disorder, inflammation, and a skin disorder.

42. 42. The method of claim 41, wherein the disorder is depression.

43. 42. The method of claim 41, wherein the condition is cardiac arrhythmia.

44. A method of cosmetic treatment comprising administering to a subject in need of cosmetic treatment a pharmaceutical composition according to any one of claims 1 to 39.

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