Recombinant polypeptide, composition comprising same, uses thereof, and methods therefor

Abstract

A recombinant polypeptide of the present invention comprises at least one of a recombinant receptor-binding domain of botulinum toxin type A1, a recombinant light chain domain of botulinum toxin type A, and a recombinant translocation domain of botulinum toxin type A. Also, the present invention provides a recombinant botulinum toxin type A comprising at least one of the above-described recombinant polypeptides and having increased potency, safety, or half-life compared to wild-type botulinum toxin.

Description

Title of Invention: RECOMBINANT POLYPEPTIDE, COMPOSITION COMPRISING SAME, USES THEREOF, AND METHODS THEREFOR Technical Field The present disclosure relates to recombinant polypeptides, and more particularly, to recombinant polypeptides including: a receptor-binding domain of botulinum toxin type A1 including one or more amino acid mutations; or a recombinant botulinum toxin type A light chain and a recombinant receptor-binding domain of botulinum toxin type A1. Background Art Botulinum toxin (BoNT) is a neurotoxic protein produced by Clostridium botulinum and related species. Botulinum toxin prevents the release of acetylcholine, which is a neurotransmitter secreted from axon terminals at neuromuscular junctions. Botulinum toxin is synthesized as a 150 kDa dichain protein consisting of a 100 kDa heavy chain and a 50 kDa light chain linked by a disulfide bond. The heavy chain is further divided into an N-terminal domain (Hn), which assists in translocation of the light chain into the cytosol of a cell, and a C-terminal domain (Hc), which recognizes and binds to cell surface receptors on nerve cells. Once inside the cell, the light chain inactivates neurotransmitter release by specifically proteolytically cleaving a portion of the soluble NSF-attachment protein receptor (SNARE). Botulinum toxin is divided into seven serotypes (botulinum toxin type A, botulinum toxin type B, botulinum toxin type C, botulinum toxin type D, botulinum toxin type E, botulinum toxin type F, and botulinum toxin type G), which are further subdivided into subtypes based on variations in amino acid sequences. Among these, botulinum toxin type A1 has been extensively studied at the molecular level and in preclinical and clinical-stage studies and is currently widely used in the pharmaceutical industry. In contrast, other botulinum toxin type A subtypes have been less studied or reported with respect to their pharmacological characteristics due to difficulties in isolating purified toxins. Currently, only some of the botulinum toxin type A subtypes have been characterized biochemically, at the cellular level, and in vivo. According to reported studies, various in vitro and in vivo studies on botulinum toxin type A subtypes have revealed characteristics that distinguish them from botulinum toxin type A1 in terms of mechanisms such as potency, intracellular trafficking, and duration of action. Recently, in vitro characteristics of botulinum toxin type A7 and A8 have been reported. Disclosure of Invention Technical Problem An object of the present disclosure is to provide a recombinant polypeptide including one or more of a recombinant receptor-binding domain and a recombinant translocation domain of botulinum toxin type A1, and a recombinant botulinum toxin including the same. An object of the present disclosure is to provide a composition related to a recombinant polypeptide including a recombinant receptor-binding domain and a recombinant translocation domain of botulinum toxin type A1, and a recombinant botulinum toxin including the same. An object of the present disclosure is to provide a pharmaceutical use of the composition. An object of the present disclosure is to provide a method for improving, preventing, or treating a disease or condition, including administering the composition to a subject. Solution to Problem A recombinant polypeptide according to an embodiment of the present disclosure may include a receptor-binding domain of botulinum toxin type A1. The receptor-binding domain may include amino acid mutations at one or more positions at which the amino acids are conserved as common amino acids in botulinum toxin type A2 and botulinum toxin type A6, but different amino acids are present at the corresponding positions in botulinum toxin type A1. The positions of the amino acid mutations may include at least one selected from the group consisting of N880, W1068, and N1090. The receptor-binding domain may further include an amino acid mutation at one or more positions selected from the group consisting of N954, M968, T990, Q991, N1025, N1026, N1052, T1063, H1064, Y1117, T1232, and L1278. As used herein, for example, an amino acid mutation at position N954 means that asparagine, which is amino acid number 954, is substituted with another amino acid, deleted, or modified. The receptor-binding domain may further include at least one amino acid mutation selected from the group consisting of N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, Y1117F, T1232R, and L1278F. The receptor-binding domain may further include an amino acid mutation at one or more positions selected from the group consisting of N954, M968, T990, Q991, N1025, N1026, N1052, T1232, and L1278. The receptor-binding domain may include at least one amino acid mutation selected from the group consisting of N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, and L1278F. The recombinant polypeptide of an embodiment may further include a translocation domain, wherein the translocation domain may include a polypeptide selected from the group consisting of a translocation domain of botulinum toxin type A1, a translocation domain of botulinum toxin type A2, a translocation domain of botulinum toxin type A6, and variants thereof. For example, the translocation domain may include or consist of a translocation domain of botulinum toxin type A1 or a variant thereof. The recombinant polypeptide of an embodiment may include a translocation domain of botulinum toxin type A1, wherein the translocation domain may include an amino acid mutation at one or more positions among positions that are conserved as common amino acids in botulinum toxin type A2 and botulinum toxin type A6 but have different amino acids at the corresponding positions in botulinum toxin type A1. Alternatively, the recombinant polypeptide of an embodiment may include a translocation domain of botulinum toxin type A1 and may include an amino acid mutation at position 589. The recombinant polypeptide of an embodiment may include a translocation domain of botulinum toxin type A1, wherein the position of the amino acid mutation in the translocation domain may include at least one selected from the group consisting of D589, L681, D696, I713, K730, E734, K779, N789, G804, E809, L815, K816, and A818. The translocation domain may include at least one amino acid mutation selected from the group consisting of D589K, E734K, and E809K. The recombinant polypeptide of an embodiment may further include a light chain domain, wherein the light chain domain may include a polypeptide selected from the group consisting of botulinum toxin type A1, botulinum toxin type A4, and variants thereof. The light chain domain may include a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of the botulinum toxin type A4 light chain, or variants thereof. The variant light chain may include a first domain and a second domain of the botulinum toxin type A1 light chain; and a third domain and a fourth domain of the botulinum toxin type A4 light chain, wherein leucine at position 260 is substituted with phenylalanine, and isoleucine at position 264 is substituted with arginine. In an embodiment, the receptor-binding domain may have a half-maximal inhibitory concentration value, according to a botulinum toxin heavy chain competition assay, that is lower than that of a wild-type receptor-binding domain. A recombinant polypeptide according to an embodiment of the present disclosure may further include a light chain domain and a translocation domain, and may have increased potency, safety, or half-life compared to wild-type botulinum toxin type A. The recombinant polypeptide may include at least one selected from recombinant botulinum toxins of SEQ ID NOs: 8 to 32, SEQ ID NO: 43, and SEQ ID NO: 44. According to an embodiment of the present disclosure, the recombinant polypeptide includes a receptor-binding domain of botulinum toxin type A1, and the receptor-binding domain may include amino acid mutations at two or more positions selected from N954, Y1117, T1232, R1273, and L1278. The receptor-binding domain may include two or more amino acid mutations selected from N954S, Y1117F, T1232R, R1273K, and L1278F. The receptor-binding domain may include amino acid mutations at two or more positions selected from N954, T1232, and L1278. The receptor-binding domain may include amino acid mutations at two or more positions selected from N954S, T1232R, and L1278F. The amino acid mutations of the receptor-binding domain may include N954S and T1232R; N954S and L1278F; T1232R and L1278F; T1232R and R1273K; R1273K and L1278F; N954S, T1232R, and L1278F; Y1117F, R1273K, and L1278F; or T1232R, R1273K, and L1278F. According to an embodiment of the present disclosure, the recombinant polypeptide includes a receptor-binding domain of botulinum toxin type A1, and a variant of the receptor-binding domain may have a half-maximal inhibitory concentration value, according to a botulinum toxin heavy chain competition assay, that is lower than that of a wild-type receptor-binding domain. The variant of the receptor-binding domain may include an amino acid mutation at one or more positions selected from the group consisting of N880, N954, M968, T990, Q991, N1025, N1026, N1052, T1063, H1064, W1068, N1090, Y1117, S1142, T1232, and L1278. The variant of the receptor-binding domain may include at least one amino acid mutation selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, Y1117F, S1142Y, S1142R, S1142I, S1142F, S1142H, T1232R, and L1278F. For example, the receptor-binding domain may include an amino acid mutation at position S1142. The amino acid mutation may include S1142Y, S1142R, S1142I, S1142F, or S1142H. According to an embodiment of the present disclosure, the recombinant botulinum toxin includes a variant in which one or more amino acids of the receptor-binding domain sequence of botulinum toxin type A1 are mutated, wherein the amino acid mutation includes one or more selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, Y1117F, S1142Y, S1142R, S1142I, S1142F, S1142H, T1232R, and L1278F, and, when administered to a subject in need of treatment, one or more of potency and safety may be superior as compared with wild-type botulinum toxin type A1, while the remainder may be at least equivalent to that of the wild-type toxin. The recombinant botulinum toxin may include a light chain domain selected from the group consisting of the aforementioned botulinum toxin type A1 light chain, botulinum toxin type A4 light chain, and variants thereof. In addition, the recombinant botulinum toxin may further include the aforementioned translocation domain. Compared with wild-type botulinum toxin type A1, the recombinant botulinum toxin may exhibit a better value for one or more of a safety margin upon administration to a subject and digit abduction score (DAS)auc or compound muscle action potential (CMAP)aac, while the remainder may be at least equivalent to that of the wild-type toxin. For example, compared with wild-type botulinum toxin type A1, the recombinant botulinum toxin may exhibit a higher value for one or more of a safety margin upon administration to a subject and DASauc or CMAPaac, while the remainder may be at least equivalent to that of the wild-type toxin. According to an embodiment of the present disclosure, the recombinant botulinum toxin may include a variant of a botulinum toxin type A light chain and a receptor-binding domain including one or more amino acid mutations selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, S1142Y, S1142R, S1142I, S1142F, S1142H, Y1117F, T1232R, and L1278F. The receptor-binding domain may include a recombinant polypeptide of SEQ ID NOs: 1 to 7, 33 to 42, and 45 to 49. The recombinant botulinum toxin may include a light chain domain selected from the group consisting of the aforementioned botulinum toxin type A1 light chain, botulinum toxin type A2 light chain, and variants thereof. Upon administration to a subject in need of treatment, the recombinant botulinum toxin may be superior in one or more of safety and duration of action compared with wild-type botulinum toxin type A1, with the remainder being at least equivalent to that of the wild-type toxin. Compared with wild-type botulinum toxin type A1, the recombinant botulinum toxin may exhibit a higher value for one or more of a safety margin upon administration to a subject and DASauc or CMAPaac. According to an embodiment of the present disclosure, the recombinant polypeptide may include a botulinum toxin type A1 translocation domain. The translocation domain may include an amino acid mutation at one or more positions where the amino acid is conserved as a common amino acid in botulinum toxin type A2 and botulinum toxin type A6, but the corresponding position in the botulinum toxin type A1 has a different amino acid. The position of the amino acid mutation in the translocation domain may include at least one selected from the group consisting of L681, D696, I713, K730, E734, K779, N789, G804, E809, L815, K816, and A818. The translocation domain may include at least one amino acid mutation selected from the group consisting of L681I, D696N, I713T, K730E, E734K, K779S, N789D, G804A, E809K, L815V, K816R, and A818V. The recombinant botulinum toxin including the translocation domain may exhibit superior specific potency compared with wild-type botulinum toxin. Advantageous Effects of Invention According to the present disclosure, a recombinant polypeptide including botulinum toxin type A1 or a subdomain thereof is provided. Brief Description of Drawings FIG. 1 is a graph of the thermal denaturation midpoint temperature (Tm) of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 10, 11, 12, 13, 14, 16, and 20. FIG. 2 is a graph showing changes in digit abduction scores (DAS) over time after administration of a botulinum toxin type A product (BoNT / A) and a recombinant botulinum toxin of SEQ ID NO: 10 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg. DAS values on each measurement day were analyzed using the Mann-Whitney test (*p<0.05, **p<0.01). FIG. 3 is a graph showing changes in DAS scores over time after administration of a botulinum toxin type A product (BoNT / A) and a recombinant botulinum toxin of SEQ ID NO: 11 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg. DAS values on each measurement day were analyzed using the Mann-Whitney test (ap = 0.065, bp = 0.093). FIG. 4 is a graph (A) showing changes in DAS scores over time after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 10 and 11 to the right gastrocnemius muscle of rats at a dose of 12 U / kg, and a graph (B) showing compound muscle action potential (CMAP) values in the left gastrocnemius muscle on day 7 after administration. DAS values on each measurement day were analyzed using the Mann-Whitney test, and CMAP values were analyzed using a two-tailed t-test (*p<0.05, ap = 0.065). FIG. 5 is a graph showing changes in DAS scores over time after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 22 and 23 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg. DAS values on each measurement day were analyzed using the Mann-Whitney test (*p<0.05, ap = 0.065, bp = 0.082). FIG. 6 is a graph showing CMAP values at an administered site (ipsilateral) and a non-administered site (contralateral) on day 7 after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 22 and 23 to the right gastrocnemius muscle of mice at a dose of 12 U / kg. CMAP values of the botulinum toxin type A product and recombinant botulinum toxins were analyzed using a two-tailed t-test according to the administration site (*p<0.05, **p<0.01). FIG. 7 is a graph showing changes in DAS scores over time after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 14 and 15 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg. DAS values on each measurement day were analyzed using the Mann-Whitney test (*p<0.05, **p<0.01, ap = 0.065, bp = 0.093). FIG. 8 is a graph showing CMAP values at an administered site (ipsilateral) and a non-administered site (contralateral) on day 7 after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 14 and 15 to the right gastrocnemius muscle of mice at a dose of 12 U / kg. CMAP values of the botulinum toxin type A product and recombinant botulinum toxins were analyzed using a two-tailed t-test according to the administration site (*p<0.05, **p<0.01, ***p<0.001). FIG. 9 is a graph showing changes in DAS scores over time after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 12 and 13 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg. DAS values on each measurement day were analyzed using the Mann-Whitney test (ap = 0.065). FIG. 10 is a graph showing CMAP values at an administered site (ipsilateral) and a non-administered site (contralateral) at 1 week (day 7) after administration of a botulinum toxin type A product (BoNT / A) and recombinant botulinum toxins of SEQ ID NOs: 12 and 13 to the right gastrocnemius muscle of mice at a dose of 12 U / kg. CMAP values of the botulinum toxin type A product and recombinant botulinum toxins were analyzed using a two-tailed t-test according to the administration site (*p<0.05). FIG. 11 is a graph showing changes over time in DAS values after administration of a botulinum toxin type A product (BoNT / A) and a recombinant botulinum toxin of SEQ ID NO: 16 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg. DAS values on each measurement day were analyzed using the Mann-Whitney test (*p<0.05, ap = 0.065, bp = 0.093). FIG. 12 is a graph showing CMAP values at the administered site (ipsilateral) and the non-administered site (contralateral) at 1 week (Day 6) after administration of a botulinum toxin type A product (BoNT / A) and a recombinant botulinum toxin of SEQ ID NO: 16 to the right gastrocnemius muscle of mice at a dose of 12 U / kg. CMAP values of the botulinum toxin type A product and recombinant botulinum toxin were analyzed using a two-tailed t-test according to the administration site (**p<0.01). Mode for Carrying out the Invention Unless otherwise defined, technical and scientific terms used herein have the same meanings as those commonly used in the field to which the present technology belongs. For an understanding of the present disclosure, the following definitions shall apply, and singular expressions include plural expressions unless the context clearly indicates otherwise, and vice versa. The term "and / or" includes all combinations that the associated components may define. In interpreting components, they shall be construed as including a margin of error even in the absence of a separate explicit description. As used herein, the term "about" is used to encompass conventional tolerance ranges in the relevant art. For example, the term "about" may mean that there is a tolerance of up to 5%, 10%, 15%, or 20% in the relevant value or range of values. As used herein, the terms "protein" and "polypeptide" are used interchangeably to refer to polymers of amino acid residues and variants and synthetic analogs thereof. These terms apply to amino acid polymers in which one or more amino acid residues is a synthetic non-naturally occurring amino acid, for example, a chemical analog of a related naturally occurring amino acid, as well as to naturally occurring amino acid polymers. These terms also include post-translational modifications of polypeptides, for example, glycosylation, phosphorylation, and acetylation. The term "botulinum toxin (BoNT)" encompasses any polypeptide or fragment of botulinum toxin. In an embodiment, botulinum toxin refers to full-length botulinum toxin or botulinum toxin-derived fragments. In an embodiment, botulinum toxin may enter neurons and carry out the overall cellular mechanism of inhibiting neurotransmitter release. Unless otherwise specifically defined herein, "botulinum toxin" includes all serotypes and variants or fusion proteins thereof. For example, the botulinum toxin may be selected from the group consisting of types A (BoNT / A), B (BoNT / B), C (BoNT / C), D (BoNT / D), E (BoNT / E), F (BoNT / F), G (BoNT / G), H (BoNT / H), and X (BoNT / X), Enterococcus species botulinum toxin J (eBoNT / J), and a mosaic botulinum toxin, and / or variants thereof. Examples of mosaic toxins include BoNT / DC, BoNT / CD, and BoNT / FA. For example, type A may be used. In an embodiment, the botulinum toxin is derived from various BoNT / A subtypes, for example, A1, A2, A3, A4, A5, A6, A7, A9, and A10; BoNT / B subtypes, for example, B1, B2, B3, B4, B5, B6, B7, B8, Bnp, and Bbv; BoNT / C subtypes, for example, C and CD; BoNT / D subtypes, for example, D and DC; BoNT / E subtypes, for example, E1, E2, E3, E4, E5, E6, E7, E8, and E9; BoNT / F subtypes, for example, F1, F2, F3, F4, F5, F6, and F7; and BoNT / G subtypes, for example, subtype G. BoNT subtypes include chimeric BoNTs, for example, BoNT / DC, BoNT / CD, BoNT / FA, and the like. As used herein, unless otherwise specified, a recombinant botulinum toxin according to an embodiment or each domain constituting the recombinant botulinum toxin (for example, a light chain, a heavy chain, a subdomain of the light chain, a translocation domain of the heavy chain, a receptor-binding domain of the heavy chain, and the like) may be a recombinant polypeptide derived from any serotype. In an embodiment, the term "variant" of botulinum toxin (including both wild-type toxins and recombinant toxins) may refer to a botulinum toxin having at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or at least 99.8% homology with the sequence of a reference botulinum toxin. For example, the term "variant" of botulinum toxin (including both wild-type toxins and recombinant toxins) may refer to a botulinum toxin having 1 to 10, 1 to 7, 1 to 5, 1 to 3, or 1 amino acid variation(s) in the sequence of a reference botulinum toxin, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid variations. In an embodiment, amino acid variations include both conservative and non-conservative variations and may include, for example, conservative variations in which an amino acid is varied while the properties of the varied amino acid (hydrophilicity, hydrophobicity, and the like) are retained. For example, a recombinant polypeptide of the present disclosure may include a variant of botulinum toxin type A1 as a receptor-binding domain; however, when the recombinant polypeptide according to an embodiment includes a light chain (and each domain thereof) and / or a translocation domain, these may include botulinum toxin type A1, serotypes other than type A1, and variants thereof. A recombinant botulinum toxin according to an embodiment may be a noncomplex botulinum toxin that does not contain complex proteins. For example, the noncomplex botulinum toxin may not contain NTNH or hemagglutinin, in which case the molecular weight of the non-complex botulinum toxin may be about 150 kDa. As used herein, the non-complex botulinum toxin may be referred to as a 7S botulinum toxin or a botulinum toxin having a molecular weight of 150 kDa. A recombinant botulinum toxin according to an embodiment may also be a complex botulinum toxin composition containing complex proteins. For example, the complex botulinum toxin may contain NTNH or hemagglutinin, in which case the molecular weight of the complex botulinum toxin may be about 900 kDa or 500 kDa. A recombinant botulinum toxin composition according to an embodiment may be a composition that does not contain animal proteins. For example, a recombinant botulinum toxin composition according to an embodiment may not contain a protein stabilizer derived from animals. In an embodiment, the recombinant botulinum toxin composition may not include albumin, such as human serum albumin or recombinant human albumin. As used herein, the terms "unit," "unit(s)," or "U" refer to an LD50 dose, defined as the amount of botulinum toxin that kills 50% of mice injected with botulinum toxin, and are used interchangeably within a single product. "Pharmaceutical composition" means a preparation containing an active ingredient. The term "preparation" means that, in addition to an active ingredient (e.g., botulinum toxin), at least one additional ingredient, such as albumin (e.g., human serum albumin or recombinant human albumin) and / or sodium chloride, is present in the pharmaceutical composition. A pharmaceutical composition is a preparation suitable for administration to a subject, such as a human patient. The pharmaceutical composition may be in lyophilized form, for example, a solution formed after reconstitution of the lyophilized pharmaceutical composition using saline or water, or in the form of a solution that does not require reconstitution. The pharmaceutical composition may be liquid or solid. The pharmaceutical composition may be free of animal-derived proteins and / or albumin. The term "active ingredient" refers to a substance in a biologically or physiologically active preparation or composition. In particular, in the context of a pharmaceutical preparation or composition, the term "active ingredient" refers to a constituent substance that exhibits the intended pharmaceutical effect. For example, the active ingredient of the present disclosure may be a non-complexed botulinum toxin free of complexing proteins. "Administration" or "administering" means the step of providing a pharmaceutical composition or active ingredient to a subject. The pharmaceutical composition may be administered through various suitable routes. In one aspect, administration of the pharmaceutical composition may be accomplished by transdermal, subcutaneous, or intramuscular administration. In an embodiment, it may be accomplished by locally administering the composition to a muscle or group of muscles. For example, reduction of forehead wrinkles and skin wrinkles may be accomplished by administering the composition transdermally or subcutaneously to the relevant wrinkles. For example, the composition of an embodiment may be administered by subcutaneous or intramuscular injection. The composition according to an embodiment may be administered by subcutaneous or intramuscular injection via a prefilled syringe. The composition of an embodiment may be administered transdermally using a transdermal patch or microneedles. "Pharmaceutical composition" refers to a preparation containing an active ingredient. The "pharmaceutical composition" may include, in addition to an active ingredient (e.g., botulinum toxin), at least one additional ingredient, such as albumin (human serum albumin or recombinant human albumin) and / or sodium chloride. The pharmaceutical composition is a preparation suitable for administration to a subject such as a mammal, including a human. The subject to which the composition herein is administered may include, without limitation, humans or animals, for example, humans, pigs, dogs, cats, cows, horses, rats, and the like. The pharmaceutical composition may be in liquid form or lyophilized form. When the pharmaceutical composition is a lyophilized preparation, the lyophilized pharmaceutical composition may be reconstituted for use using, for example, saline or water. It may be in solution form. The pharmaceutical composition may be liquid or solid. The pharmaceutical composition may be free of animal-derived proteins and / or albumin. "Treat," "treating," or "treatment" means alleviation or reduction (including partial reduction, substantial reduction, near-complete reduction, and complete reduction), resolution, or prevention (temporary or permanent) of a disease, disorder, or abnormality, so as to achieve a desired therapeutic result, for example, by healing wounded or damaged tissue, or by altering, changing, enhancing, improving, ameliorating, and / or beautifying an existing or perceived disease, disorder, or abnormality. As used herein, "treatment" includes prevention. "Prevention" means delaying the onset of a disease, disorder, or condition. Prevention may be considered complete when the onset of a disease, disorder, or condition is delayed for a predetermined period. In an embodiment, "treatment" refers to the treatment of a disease, disorder, or medical condition in a patient, such as a mammal (particularly a human), including one or more of the following: (a) preventing the occurrence of the disease, disorder, or medical condition, i.e., preventing recurrence of the disease or medical condition or prophylactically treating a patient predisposed to the disease or medical condition; (b) ameliorating the disease, disorder, or medical condition, including antagonizing the effects of other therapeutic agents, i.e., eliminating or causing regression of the disease, disorder, or medical condition in the patient; (c) suppressing the disease, disorder, or medical condition, i.e., slowing or arresting the development of the disease, disorder, or medical condition in the patient; or (d) alleviating the symptoms of the disease, disorder, or medical condition in the patient. As used herein, the terms "unit," "unit(s)," or "U" refer to an LD50 dose, defined as the amount of botulinum toxin that kills 50% of mice injected with botulinum toxin, and are used interchangeably within a product. In one aspect, a therapeutically effective amount of botulinum toxin is about 0.01 U / kg to about 100 U / kg, about 0.1 U / kg to about 100 U / kg, about 0.2 U / kg to about 100 U / kg, about 0.2 U / kg to about 50 U / kg, about 0.2 U / kg to about 30 U / kg, about 0.2 U / kg to about 10 U / kg, about 0.2 U / kg to about 1 U / kg, about 1.2 U / kg, about 4 U / kg, or about 12 U / kg. In another aspect, based on a 60 kg adult, the amount may be about 1 U to about 10,000 U, about 1 U to about 5,000 U, about 1 U to about 2,500 U, about 1 U to about 1,000 U, about 1 U to about 500 U, about 1 U to about 300 U, about 1 U to about 200 U, about 10 U to about 200 U, about 10 U to about 100 U, or about 10 U to about 50 U. In the present disclosure, "free of animal proteins" means substantially free of proteins derived from animals (e.g., human serum albumin). The composition of an embodiment may be administered in a therapeutically effective amount, and as used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount sufficient to treat or prevent a disease at a reasonable benefit / risk ratio applicable to any medical treatment or prevention. Effective dosage levels may be determined depending on the severity of the disease, the activity of the drug, the age, body weight, health, and sex of the patient, sensitivity to the drug, the timing of administration, the route of administration and rate of excretion of the composition of the present disclosure, the duration of treatment, drugs used concomitantly or in combination with the composition of the present disclosure, and other factors known in the medical field. In an embodiment, botulinum toxin may be administered in a single or multiple treatment sessions. The dosage may be administered at the injection site as a single or divided injection. In multiple treatment sessions, botulinum toxin may be administered at intervals of 6 months, 4 months, or 3 months or less. Multiple treatment sessions may include a primary treatment and a secondary treatment, and the dosage of the secondary treatment may be less than, greater than, or the same as the dosage of the primary treatment. "Botulinum toxin" may be produced by bacteria or recombinant methods. Botulinum toxin includes botulinum toxin derivatives. A botulinum toxin derivative may be a derivative including one or more chemical or functional modifications on a portion or partial chain of a natural botulinum toxin or recombinant native botulinum toxin having botulinum toxin activity. For example, a botulinum toxin derivative may be a modified toxin having one or more deleted, modified, or substituted amino acids. Botulinum toxin may be a recombinant peptide, a fusion protein, or, for example, a hybrid neurotoxin produced from subunits or domains of different toxin serotypes. Botulinum toxin may also be a portion of an entire molecule having toxin activity and may be used as part of a combination thereof or a conjugated molecule, for example, a fusion protein. The term "domain" refers to a tertiary structure that can evolve, function, and exist independently of the remainder of the protein chain, with a given protein sequence and conserved portion. Because domains are independently stable, domains may be domain-swapped by genetic engineering between one protein and another protein to generate a chimeric protein. The term "homology" refers to the degree of amino acid sequence identity between polypeptides. For example, when a first amino acid sequence is identical to a second amino acid sequence, the first and second amino acid sequences exhibit 100% homology. The term "recombination" refers to a process in which gene-constituting elements, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), are rearranged into a sequence different from the original sequence during disassembly and reassembly. Through molecular biology experiments, fragments of DNA may be artificially recombined. Such artificially recombined DNA is referred to as recombinant DNA. A recombinant polypeptide according to an embodiment of the present disclosure may include a receptor-binding domain (RBD) of botulinum toxin type A1. The receptorbinding domain may include an amino acid mutation. The receptor-binding domain may include an amino acid mutation at one or more positions that are conserved as amino acids common to botulinum toxin type A2 and botulinum toxin type A6, but have different amino acids at the corresponding positions in the botulinum toxin type A1. For example, the amino acid mutation may be one in which at least a portion of the sequence of botulinum toxin type A1 is substituted with an amino acid sequence common to botulinum toxin type A2 and A6. For example, asparagine, which is amino acid residue 880 of the receptor-binding domain of botulinum toxin type A1, may be substituted with serine, which is amino acid residue 880 of botulinum toxin type A2 and botulinum toxin type A6. For example, the amino acid mutation may be one in which at least a portion of the amino acid sequence of botulinum toxin type A1 is substituted with an amino acid other than an amino acid common to botulinum toxin type A2 and A6. For example, asparagine, which is amino acid residue 880 of the receptor-binding domain of botulinum toxin type A1, may be substituted with an amino acid other than serine, which is amino acid residue 880 of botulinum toxin type A2 and botulinum toxin type A6 (for example, a natural amino acid, a stereoisomer thereof, or a variant thereof). For example, asparagine at position 880 of the receptor-binding domain of botulinum toxin type A1 may be conservatively substituted with another amino acid, for example, threonine or glutamine. The recombinant polypeptide of an embodiment may include at least one amino acid mutation selected from the group consisting of N880, W1068, and N1090, for example, one, two, or three amino acid mutations. For example, the receptor-binding domain may include at least one amino acid mutation selected from the group consisting of N880S, W1068M, and N1090S, for example, one, two, or three amino acid mutations. Each amino acid mutation may be independently selected from deletion, modification, and substitution. For example, the amino acid mutation may be a substitution. The receptor-binding domain may further include at least one amino acid mutation selected from the group consisting of N954, M968, T990, Q991, N1025, N1026, N1052, T1063, H1064, Y1117, T1232, and L1278, for example, one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve amino acid mutations. For example, the receptor-binding domain may further include at least one amino acid mutation selected from the group consisting of N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, Y1117F, T1232R, and L1278F, for example, one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve amino acid mutations. The receptor-binding domain may further include at least one amino acid mutation selected from the group consisting of N954, M968, T990, Q991, N1025, N1026, N1052, T1232, and L1278, for example, one, two, three, four, five, six, seven, eight, or nine amino acid mutations. For example, the receptor-binding domain may include at least one amino acid mutation selected from the group consisting of N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, and L1278F, for example, one, two, three, four, five, six, seven, eight, or nine amino acid mutations. For example, the receptor-binding domain may include N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, and L1278F, and may include at least one or two or more amino acid mutations selected from the group consisting of T1063P, H1064R, and Y1117F. For example, the receptor-binding domain may include amino acid mutations of N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, and L1278F, and may include an amino acid mutation of "T1063P and H1064R" or "Y1117F." The receptor-binding domain may further include amino acid mutations at one or more positions other than positions at which the amino acids are conserved in both botulinum toxin type A2 and botulinum toxin type A6 but differ at the corresponding positions in botulinum toxin type A1. For example, the receptor-binding domain may further include an amino acid mutation at position S1142. For example, the amino acid mutation may further include S1142Y, S1142R, S1142I, S1142F, or S1142H. The positions of the additional nine mutable amino acids were also selected from sequences that are common to botulinum toxin types A2 and A6 and differ from the sequence of botulinum toxin type A1. In an embodiment, the receptor-binding domain may include all of the amino acid mutations of N880, W1068, and N1090, and N954, M968, T990, Q991, N1025, N1026, N1052, T1232, and L1278. In an embodiment, the receptor-binding domain may include all of the amino acid sequence mutations of N880S, W1068M, and N1090S, and N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, and L1278F. In an embodiment, the receptor-binding domain may include one of SEQ ID NOs: 1 to 7 and 33 to 42. The amino acid mutations may each independently be selected from deletion, modification, and substitution. For example, the amino acid mutation may be a substitution. A recombinant polypeptide according to an embodiment of the present disclosure may include a translocation domain of recombinant botulinum toxin type A1. The recombinant botulinum toxin type A1 translocation domain may include an amino acid mutation at one or more positions at which the amino acid is conserved as a common amino acid in botulinum toxin type A2 and botulinum toxin type A6, but at which the corresponding position in the botulinum toxin type A1 has a different amino acid. For example, the positions of amino acid mutations in the recombinant translocation domain of an embodiment may include at least one selected from the group consisting of L681, D696, I713, K730, E734, K779, N789, G804, E809, L815, K816, and A818. For example, the recombinant translocation domain of an embodiment may include at least one amino acid mutation selected from the group consisting of L681I, D696N, I713T, K730E, E734K, K779S, N789D, G804A, E809K, L815V, K816R, and A818V. Alternatively, the recombinant translocation domain of an embodiment may include a translocation domain of botulinum toxin type A1 and may include at least one amino acid mutation at a position selected from the group consisting of D589, E734, and E809. For example, the recombinant translocation domain of an embodiment may include a translocation domain of botulinum toxin type A1 and may include at least one amino acid mutation at a position selected from the group consisting of D589K, E734K, and E809K. In an embodiment, the translocation domain may include or consist of a translocation domain of wild-type botulinum toxin type A1, or may include or consist of a translocation domain of botulinum toxin type A1 including at least one amino acid mutation selected from the group consisting of D589K, L681I, D696N, I713T, K730E, E734K, K779S, N789D, G804A, E809K, L815V, K816R, A818V, E734K, and E809K. In an embodiment of the present disclosure, the recombinant polypeptide (e.g., the recombinant receptor-binding domain) may further include a translocation domain. The translocation domain may include a polypeptide selected from the group consisting of a translocation domain of botulinum toxin type A1, a translocation domain of botulinum toxin type A2, a translocation domain of botulinum toxin type A6, and variants thereof (e.g., the recombinant translocation domain of recombinant botulinum toxin type A1 described above). For example, the translocation domain may include a polypeptide selected from the group consisting of a translocation domain of botulinum toxin type A2, a translocation domain of botulinum toxin type A6, and variants thereof. For example, the translocation domain may include a translocation domain of botulinum toxin type A2 or a translocation domain of botulinum toxin type A6. The recombinant polypeptide of an embodiment may include the recombinant receptor-binding domain described above and / or the recombinant translocation domain described above. In an embodiment, the recombinant polypeptide includes the receptorbinding domain of botulinum toxin type A1 described above The recombinant polypeptide of an embodiment may further include a light chain domain. The light chain domain may be a recombinant polypeptide. For example, the light chain domain may include a polypeptide selected from the group consisting of botulinum toxin type A1, botulinum toxin type A4, and variants thereof. For example, the light chain domain may include a subdomain of botulinum toxin type A1 or a variant thereof, or a subdomain of botulinum toxin type A4 or a variant thereof. A variant of each subdomain of botulinum toxin type A1 or A4 may have at least 99.8%, at least 99.5%, at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 95%, at least 93%, at least 92%, at least 91%, or at least 90% homology with the sequence of each subdomain of wild-type botulinum toxin type A1 or A4. For example, a variant of each subdomain of botulinum toxin type A1 or A4 may include one, two, three, four, five, six, seven, eight, or nine amino acid mutations in the sequence of each subdomain of wild-type botulinum toxin type A1 or A4. In an embodiment, the sequences of the light chains of botulinum toxin type A1 and A4 were divided into first, second, third, and fourth domains according to the criteria below. The first domain of the botulinum toxin type A1 light chain and the first domain of the botulinum toxin type A4 light chain are defined as regions mainly interacting with the substrate SNAP25 and including an a-exosite. The first domain of the botulinum toxin type A1 and A4 light chains may include amino acids at positions 1 to 20 through amino acids at positions 90 to 150, for example, amino acids at positions 1, 5, 10, 15, or 20 through amino acids at positions 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 150. In an embodiment, the first domain of the light chain of botulinum toxin type A1 and the first domain of the botulinum toxin type A4 light chain include regions interacting with the substrate SNAP25 as regions including an a-exosite, and may include amino acids at positions 1, 5, 10, 15, or 20 through amino acids at positions 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, or 150, for example, amino acids at positions 20 to 90, amino acids at positions 20 to 100, amino acids at positions 20 to 110, amino acids at positions 20 to 120, amino acids at positions 20 to 130, amino acids at positions 20 to 140, amino acids at positions 10 to 90, amino acids at positions 10 to 100, amino acids at positions 10 to 110, amino acids at positions 10 to 120, amino acids at positions 10 to 130, or amino acids at positions 10 to 140. For example, the first domain of the light chain of botulinum toxin type A1 and the first domain of the botulinum toxin type A4 light chain may include amino acids at positions 1 to 120. The second domain of the botulinum toxin type A1 light chain and the second domain of the botulinum toxin type A4 light chain are defined as regions mainly involved in the enzymatic activity of the light chain, including the 170 loop, and binding to zinc as a cofactor. The second domains of the botulinum toxin type A1 light chain and the botulinum toxin type A4 light chain may include amino acids at positions 91 to 151, for example, amino acids at positions 91, 96, 101, 106, 111, 116, 121, 126, 131, 136, 141, 146, and 151, through amino acids at positions 210 to 270, for example, amino acids at positions 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, and 270. In an embodiment, the second domain of the botulinum toxin type A1 light chain and the second domain of the botulinum toxin type A4 light chain are involved in the enzymatic activity of the light chain, include the 170 loop, include regions binding to zinc as a cofactor, and may include amino acids at positions 91, 96, 101, 106, 111, 116, 121, 126, 131, 136, 141, 146, and 151 through amino acids at positions 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, and 270, for example, amino acids at positions 91 to 270, amino acids at positions 101 to 260, amino acids at positions 111 to 250, amino acids at positions 121 to 240, amino acids at positions 131 to 230, amino acids at positions 141 to 220, or amino acids at positions 151 to 210. For example, the second domain of the light chain of botulinum toxin type A1 and the second domain of the botulinum toxin type A4 light chain may include amino acids at positions 121 to 240. The third domain of the botulinum toxin type A1 light chain and the third domain of the botulinum toxin type A4 light chain are defined as domains including the 250 loop and the 370 loop, which stabilize substrate binding, and regions involved in both the a-exosite and p-exosite, which are substrate-binding sites. The third domain of the botulinum toxin type A1 and A4 light chains may include amino acids 211 to 271 (for example, amino acids 211, 216, 221, 226, 231, 236, 241, 246, 251, 256, 261, 266, and 271) to amino acids 386 to 446 (for example, amino acids 386, 391, 396, 401, 406, 411, 416, 421, 426, 431, 436, 441, and 446). The third domain of the botulinum toxin type A1 light chain and the third domain of the botulinum toxin type A4 light chain include the 250 loop and the 370 loop, which stabilize substrate binding, and regions involved in both the a-exosite and p-exosite, which are substrate-binding sites, and may include amino acids 211, 216, 221, 226, 231, 236, 241, 246, 251, 256, 261, 266, and 271 to amino acids 386, 391, 396, 401, 406, 411, 416, 421, 426, 431, 436, 441, and 446, for example, amino acids 211 to 446, amino acids 221 to 436, amino acids 231 to 426, amino acids 241 to 416, amino acids 251 to 406, or amino acids 261 to 396. For example, the third domain of the light chain of botulinum toxin type A1 and the third domain of the botulinum toxin type A4 light chain may include amino acids at positions 241 to 396. The fourth domain of the botulinum toxin type A1 light chain (SEQ ID NO: 4) and the fourth domain of the botulinum toxin type A4 light chain (SEQ ID NO: 8) may be defined as domains including regions involved in the structural flexibility of the botulinum toxin type A light chain. The fourth domain of the botulinum toxin type A1 light chain and the fourth domain of the botulinum toxin type A4 light chain may include amino acids 362 to 425 (for example, amino acids 362, 367, 372, 377, 382, 387, 392, 397, 402, 407, 412, 417, 422, and 425) to amino acids 410 to 448 (for example, amino acids 410, 414, 417, 420, 425, 430, 435, 440, 445, and 448). In an embodiment, the fourth domain of the botulinum toxin type A1 light chain (SEQ ID NO: 4) and the fourth domain of the botulinum toxin type A4 light chain (SEQ ID NO: 8) may include regions involved in the structural flexibility of the botulinum toxin type A light chain and may include amino acids 362, 367, 372, 377, 382, 387, 392, 397, 402, 407, 412, 417, 422, and 425 to amino acids 410, 414, 417, 420, 425, 430, 435, 440, 445, and 448, for example, amino acids 362 to 448, amino acids 372 to 438, or amino acids 382 to 428. For example, the fourth domain of the light chain of botulinum toxin type A1 and the fourth domain of the botulinum toxin type A4 light chain may include amino acids at positions 397 to 425. The light chain domain may be one in which, among the first, second, third, and fourth domains of a botulinum toxin type A light chain other than botulinum toxin type A4, the sequence of the third domain is substituted with the sequence of the third domain of botulinum toxin type A4 or a variant thereof. The variant of the third domain of botulinum toxin type A4 may have 99% or more, 98% or more, 97% or more, 96% or more, 95% or more, 95% or more, 93% or more, 92% or more, 91% or more, or 90% or more homology with the sequence of the third domain of wild-type botulinum toxin type A4. For example, the variant of the third domain of botulinum toxin type A4 may include one, two, three, four, five, six, seven, eight, or nine amino acid mutations in the sequence of the third domain of wild-type botulinum toxin type A4. Here, each amino acid mutation may independently be selected from deletion, modification, and substitution, and, for example, the amino acid mutation may be an amino acid substitution. The substantially same contents as those described for the variant of the third domain of botulinum toxin type A4 may be applicable to a variant of the fourth domain of botulinum toxin type A4, a variant of the first domain of botulinum toxin type A1, a variant of the second domain of botulinum toxin type A1, and a variant of the fourth domain of the botulinum toxin type A1 light chain, which are described below. For example, these variants may have 99% or more, 98% or more, 97% or more, 96% or more, 95% or more, 95% or more, 93% or more, 92% or more, 91% or more, or 90% or more homology with the sequence of the wild-type domain, and may include one, two, three, four, five, six, seven, eight, or nine amino acid mutations in the sequence of the wild-type domain. The light chain domain may be one in which the sequence of the fourth domain among the first, second, third, and fourth domains of the recombinant botulinum toxin type A light chain is further substituted with the sequence of the fourth domain of botulinum toxin type A4 or a variant thereof. The light chain domain may be one in which the sequence of the second domain among the first, second, third, and fourth domains of a botulinum toxin type A light chain other than botulinum toxin type A1 is substituted with the sequence of the second domain of botulinum toxin type A1 or a variant thereof. The light chain domain may be one in which the sequence of the fourth domain among the first, second, third, and fourth domains of the recombinant botulinum toxin type A light chain is further substituted with the sequence of the fourth domain of botulinum toxin type A1 or a variant thereof. In an embodiment, the light chain domain may include the first domain and the second domain of the botulinum toxin type A1 light chain; or variants thereof. The light chain domain may further include the third domain and the fourth domain of the botulinum toxin type A4 light chain; or variants thereof. In an embodiment, a variant of each of the first, second, third, or fourth domains of the botulinum toxin type A light chain may refer to a subdomain of the botulinum toxin type A light chain having 98% or more homology or 99% or more homology, respectively, with the first, second, third, or fourth domain of the wild-type botulinum toxin type A light chain. In an embodiment, each variant of each of the first, second, third, or fourth domains of the botulinum toxin type A light chain may refer to a subdomain of the botulinum toxin type A light chain having, in each of the first, second, third, or fourth domain of the wildtype botulinum toxin type A light chain, 1 to 10, 1 to 7, 1 to 5, or 1 to 3 amino acid mutations, for example, one, two, three, four, five, six, seven, eight, nine, or 10 amino acid mutations. The light chain domain of an embodiment may include a recombinant light chain disclosed in PCT-KR2022-008793 or PCT-KR2022-008798, the disclosures of which are incorporated herein by reference in their entirety. The light chain domain of an embodiment may include the first domain and the second domain of the wild-type botulinum toxin type A1 light chain, or variants thereof; and the third domain and the fourth domain of the wild-type botulinum toxin type A4 light chain, or variants thereof. When the light chain domain of an embodiment is a variant, the variant of the corresponding subdomain may have 99% or more, 98% or more, 97% or more, 96% or more, 95% or more, 94% or more, 93% or more, 92% or more, 91% or more, or 90% or more homology with the sequence of the light chain subdomain of the wild-type botulinum toxin. For example, the variant of the corresponding subdomain may include one, two, three, four, five, six, seven, eight, or nine amino acid mutations in the sequence of the light chain subdomain of the wild-type botulinum toxin. When the light chain domain of an embodiment is a variant, the third domain of the botulinum toxin type A4 light chain may be mutated. For example, a variant of the third domain of botulinum toxin type A4 may have 99% or more, 98% or more, 97% or more, 96% or more, 95% or more, 94% or more, 93% or more, 92% or more, 91% or more, or 90% or more homology with the sequence of the third domain of wild-type botulinum toxin type A4. For example, a variant of the third domain of botulinum toxin type A4 may include one, two, three, four, five, six, seven, eight, or nine amino acid mutations in the sequence of the third domain of wild-type botulinum toxin type A4. For example, a variant of the third domain of botulinum toxin type A4 may have an amino acid mutation at position 260 and / or position 264 based on the full-length light chain domain. For example, a variant of the third domain of botulinum toxin type A4 may have leucine at position 260 substituted with phenylalanine and isoleucine at position 264 substituted with arginine, based on the full-length light chain domain. The above description of the light chain domain may also apply to the light chain domains described below to the extent that there is no inconsistency. The recombinant receptor-binding domain of an embodiment may include one or more sequences selected from among SEQ ID NOs: 1 to 7, 33 to 42, and 45 to 49. The recombinant polypeptide of an embodiment may further include a cell penetrating peptide. In an embodiment, the cell penetrating peptide may be IMTP-8. The cell penetrating peptide of an embodiment may be linked to the recombinant polypeptide through a GS linker. The cell penetrating peptide may be directly or indirectly linked to the recombinant polypeptide through a GS linker. The GS linker is an amino acid linker rich in glycine and including glycine and serine, and may include conventional GS linkers used in the art, and may be a linker including four or more or five or more G and S residues. For example, the GS linker may be GGGGS. The receptor-binding domain of an embodiment may have a half-maximal inhibitory concentration value, according to a botulinum toxin heavy chain competition assay, that is lower than that of a wild-type receptor-binding domain. For example, the receptor-binding domain of an embodiment may have a half-maximal inhibitory concentration value, according to an ELISA-based botulinum toxin heavy chain competition assay, that is lower than that of a wild-type receptor-binding domain. The recombinant polypeptide of an embodiment may further include a light chain domain, wherein the light chain domain includes a polypeptide selected from the group consisting of botulinum toxin type A1, botulinum toxin type A2, and variants thereof; and may further include a translocation domain, wherein the translocation domain includes a polypeptide selected from the group consisting of a translocation domain of botulinum toxin type A1, a translocation domain of botulinum toxin type A2, a translocation domain of botulinum toxin type A6, and variants thereof. When the translocation domain of an embodiment is a variant, the variant of the translocation domain may have at least 99.5%, at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91%, at least 90%, at least 89%, at least 88%, at least 87%, at least 86%, or at least 85% homology with the sequence of a translocation domain of wild-type botulinum toxin (e.g., botulinum toxin type A1, A2, or A6). For example, a variant of the relevant subdomain may include one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, or more amino acid mutations in the sequence of the light chain subdomain of wild-type botulinum toxin, or may include thirty, twenty-five, twenty, eighteen, fifteen, or fewer amino acid mutations. The translocation domain of an embodiment may include at least one amino acid mutation selected from the group consisting of L681, D696, I713, K730, E734, K779, N789, G804, E809, L815, K816, and A818. The translocation domain of an embodiment may include at least one amino acid mutation selected from the group consisting of L681I, D696N, I713T, K730E, E734K, K779S, N789D, G804A, E809K, L815V, K816R, and A818V. The translocation domain of an embodiment may include at least one amino acid mutation selected from the group consisting of D589, E734, and E809. The translocation domain of an embodiment may include at least one amino acid mutation selected from the group consisting of D589K, E734K, and E809K. The recombinant polypeptide of an embodiment may have increased potency, safety, or half-life relative to wild-type botulinum toxin type A. The recombinant polypeptide of an embodiment may include one or more sequences selected from SEQ ID NOs: 1 to 7, 33 to 42, and 45 to 49; SEQ ID NOs: 8 to 32, 43, and 44; or SEQ ID NOs: 1 to 49 in Table 1 described below. A recombinant polypeptide according to an embodiment of the present disclosure includes a receptor-binding domain of botulinum toxin type A1, and the receptor-binding domain may include amino acid mutations at two or more positions selected from N954, Y1117, T1232, R1273, and L1278. For example, the receptor-binding domain may include two or more amino acid mutations selected from N954S, Y1117F, T1232R, R1273K, and L1278F. In an embodiment, the receptor-binding domain may include amino acid mutations at two or more positions selected from N954, T1232, and L1278. For example, the receptor-binding domain may include amino acid mutations at two or more positions selected from N954S, T1232R, and L1278F. For example, the amino acid mutations of the receptor-binding domain may include N954S and T1232R; N954S and L1278F; T1232R and L1278F; T1232R and R1273K; R1273K and L1278F; N954S, T1232R, and L1278F; Y1117F, R1273K, and L1278F; or T1232R, R1273K, and L1278F. A recombinant polypeptide according to an embodiment of the present disclosure may include a receptor-binding domain of botulinum toxin type A1. Here, the half-maximal inhibitory concentration value of a variant of the receptor-binding domain according to a botulinum toxin heavy chain competition assay may be lower than that of the wild-type receptor-binding domain. Here, the variant of the receptor-binding domain may include an amino acid mutation at one or more positions selected from the group consisting of N880, N954, M968, T990, Q991, N1025, N1026, N1052, T1063, H1064, W1068, N1090, Y1117, S1142, T1232, and L1278. For example, the receptor-binding domain may include an amino acid mutation at position S1142. For example, the amino acid mutation may include S1142Y, S1142R, S1142I, S1142F, or S1142H. A recombinant botulinum toxin according to an embodiment of the present disclosure may include a variant in which two or more amino acids of the receptor-binding domain sequence of botulinum toxin type A1 are mutated. The amino acid mutations may include two or more selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, Y1117F, S1142Y, S1142R, S1142I, S1142F, S1142H, T1232R, and L1278F. A recombinant botulinum toxin according to an embodiment may, upon administration to a subject in need of treatment, be superior in one or more of potency and safety compared to wildtype botulinum toxin type A1, with the remainder being at least equivalent to that of the wild-type toxin. For example, a recombinant botulinum toxin according to an embodiment may be superior in at least one of: compound muscle action potential (CMAP) or digit abduction score (DAS) (e.g., CMAPauc or DASaac); and body weight reduction or safety margin compared to wild-type botulinum toxin type A1, with the remainder being at least equivalent to that of the wild-type toxin. As used herein, "at least equivalent" may mean that the corresponding value of an exemplary material is not statistically significantly inferior in effect to the corresponding value of a material to be compared. A recombinant botulinum toxin according to an embodiment may include a light chain domain selected from the group consisting of a botulinum toxin type A1 light chain, a botulinum toxin type A2 light chain, and variants thereof. Here, the description of the light chain domain described above may apply equally to the light chain domain. For example, the light chain domain of a recombinant botulinum toxin according to an embodiment may include a first domain and a second domain of a botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of a botulinum toxin type A4 light chain, or variants thereof. For example, in the light chain domain of a recombinant botulinum toxin, the first domain may include a first domain of a botulinum toxin type A1 light chain or a variant thereof, the second domain may include a second domain of a botulinum toxin type A1 light chain or a variant thereof, the third domain may include a third domain of a botulinum toxin type A4 light chain or a variant thereof, and the fourth domain may include a fourth domain of a botulinum toxin type A4 light chain or a variant thereof. For example, a variant of the third domain of botulinum toxin type A4 may have a mutation in the amino acid at position 260 and / or position 264 based on the full-length light chain domain. For example, a variant of the third domain of botulinum toxin type A4 may have leucine at position 260 substituted with phenylalanine and isoleucine at position 264 substituted with arginine, based on the full-length light chain domain. A recombinant botulinum toxin according to an embodiment may, compared to wild-type botulinum toxin type A1, exhibit a superior value in one or more of safety margin upon administration to a subject; and DASauc or CMAPaac, with the remainder being at least equivalent to that of the wild-type toxin. In a recombinant botulinum toxin according to an embodiment, the translocation domain may include a polypeptide selected from the group consisting of a translocation domain of botulinum toxin type A1, a translocation domain of botulinum toxin type A2, a translocation domain of botulinum toxin type A6, and variants thereof. A translocation domain according to an embodiment may include at least one amino acid mutation selected from the group consisting of D589, E734, and E809. A translocation domain according to an embodiment may include at least one amino acid mutation selected from the group consisting of D589K, E734K, and E809K. A recombinant botulinum toxin according to an embodiment may include the aforementioned botulinum toxin type A light chain variants (e.g., variants of the A1-1, A1-2, A4-3, and A4-4 forms), and may include a receptor-binding domain including one or more amino acid mutations selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, S1142Y, S1142R, S1142I, S1142F, S1142H, Y1117F, T1232R, and L1278F. A receptorbinding domain according to an embodiment may include a recombinant polypeptide of SEQ ID NOs: 1 to 7, 33 to 42, and 45 to 49. A recombinant botulinum toxin according to an embodiment may include a translocation domain including one or more amino acid mutations selected from L260F and I264R. A variant light chain of a recombinant botulinum toxin according to an embodiment may include the first domain and the second domain of a botulinum toxin type A1 light chain, or variants thereof; and the third domain and the fourth domain of a botulinum toxin type A4 light chain, or variants thereof. A light chain domain according to an embodiment may include a variant of the second domain of a botulinum toxin type A1 light chain, wherein leucine at position 260 of the variant of the second domain of the botulinum toxin type A1 light chain is substituted with phenylalanine, and isoleucine at position 264 is substituted with arginine. A recombinant botulinum toxin according to an embodiment may exhibit inhibited diffusion upon administration to a patient, an increased duration, and rapid onset of action, compared to wild-type botulinum toxin. A recombinant botulinum toxin according to an embodiment may, compared to wild-type botulinum toxin type A1, exhibit a higher value in one or more of safety upon administration to a subject; and duration of action, with the remainder being at least equivalent to that of the wild-type toxin. For example, a recombinant botulinum toxin according to an embodiment may, compared to wild-type botulinum toxin type A1, have a higher safety margin upon administration to a subject, and DASauc or CMAPaac that is equivalent to or greater than that of the wild-type toxin. For example, a recombinant botulinum toxin according to an embodiment may, compared to wild-type botulinum toxin type A1, have an equivalent or higher safety margin upon administration to a subject, and DASauc or CMAPaac that is higher than that of the wild-type toxin. Botulinum toxins including the various recombinant polypeptides included in the present disclosure described above may have increased potency, safety, or half-life compared to wild-type botulinum toxin type A (e.g., wild-type botulinum toxin type A1). For example, a botulinum toxin including a recombinant polypeptide according to an embodiment may exhibit inhibited diffusion upon administration to a patient, an increased duration, and rapid onset of action, compared to wild-type botulinum toxin. A botulinum toxin including a recombinant polypeptide according to an embodiment may include a pharmaceutically acceptable excipient or additive. The pharmaceutically acceptable excipient or additive may be a stabilizer, an ionic compound, a surfactant, a buffer, a lyoprotectant, or a combination thereof, for example, an amino acid (e.g., methionine), a salt (e.g., NaCl), a buffer solution, a nonionic surfactant (e.g., polysorbate, such as polysorbate 20, or poloxamer), a sugar (e.g., a monosaccharide or a disaccharide such as sucrose, trehalose, and the like), a sugar alcohol (e.g., sorbitol), or a combination thereof. A botulinum toxin composition including a recombinant polypeptide of the present disclosure may be formulated in any form, such as a solid or liquid formulation, for example, a lyophilized powder, a liquid formulation, or a pre-filled syringe formulation. In an embodiment of the present disclosure, there is provided a composition for ameliorating, preventing, or treating a disease, including, as an active ingredient, a botulinum toxin including a recombinant polypeptide. In an embodiment of the present disclosure, there is provided use for ameliorating or treating a disease by administering to a subject a botulinum toxin composition including, as an active ingredient, a botulinum toxin including a recombinant polypeptide. In an embodiment of the present disclosure, there is provided a method for ameliorating, preventing, or treating a disease by administering to a subject a botulinum toxin composition including, as an active ingredient, a botulinum toxin including a recombinant polypeptide. In an embodiment of the present disclosure, there is provided use of a botulinum toxin including a recombinant polypeptide in the manufacture of a medicament for ameliorating, preventing, or treating a disease. In an embodiment, the condition or disease may include wrinkles, square jaw, pointed chin, wounds, skin softening, scars, acne, pores, elasticity, reduction of sebum secretion, and / or keloids; and / or facial spasm, eyelid spasm, torticollis, blepharospasm, cervical dystonia, mid-pharyngeal dystonia, spasmodic dysphonia, migraine, anal pruritus, and / or hyperhidrosis. In an embodiment of the present disclosure, the botulinum toxin composition including a recombinant polypeptide may include or may not include an animal-derived protein (e.g., albumin such as human serum albumin). When the composition of an embodiment does not include an animal-derived protein, it may include an amino acid as a stabilizer. In an embodiment, the amino acid may be, but is not limited to, any of the 20 standard amino acids found in nature. The amino acid of an embodiment may include any amino acid known in the art as a stabilizer. For example, in an embodiment, the amino acid may include a nonpolar amino acid, for example, methionine, isoleucine, and / or tryptophan. Hereinafter, the present disclosure will be described in more detail with reference to Examples, but these are merely for describing the present disclosure and are not intended to limit the scope thereof in any way. Example 1: Purification of a domain-substituted botulinum toxin type A heavy chain receptor-binding domain A recombinant receptor-binding domain expression vector (cloned into a pET28a vector) was introduced into BL21(DE3) cells, a type of Escherichia coli (E. coli), to induce expression of a domain-substituted recombinant botulinum toxin type A heavy chain receptor-binding domain (receptor-binding domain, RBD), i.e., the recombinant polypeptide corresponding to each embodiment of the present disclosure. Expression was induced with IPTG (isopropyl p-D-1-thiogalactopyranoside). That is, after adding IPTG to the liquid medium, BL21(DE3) cells cultured at an appropriate temperature for an appropriate period of time were centrifuged to pellet the cells. The medium in the supernatant was removed, and the pelleted cells were resuspended in A buffer (20 mM sodium phosphate, 150 mM NaCl, 10 mM imidazole, pH 7.3), followed by cell disruption on ice using a sonicator. After centrifugation of the disrupted cells, the recombinant polypeptide was isolated from the supernatant, from which cell debris had been removed, using Ni-NTA resin through binding between the His tag and the Ni-NTA resin. The amino acid sequences of the isolated recombinant polypeptides are shown in Table 1. [Table 1] SEQ ID NO. Receptor-binding domain sequence 1 A1(N880S) 2 A1(W1068M) 3 A1(N1090S) 4 A1(N880S, W1068M) 5 A1(N880S, N1090S) 6 A1(W1068M, N1090S) 7 A1(N880S, W1068M, N1090S) 33 A1(T1232R, L1278F) 34 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 35 A1 (N954S, T1232R, L1278F) 36 A1 (Y1117F, R1273K, L1278F) 37 A1 (T1232R, R1273K, L1278F) 38 A1 (S1142Y) 39 A1 (S1142R) 40 A1 (S1142I) 41 A1(S1142F) 42 A1(S1142H) 45 A1(H1064Y) 46 A1 (H1064W) 47 A1 (T1063P,H1064F) 48 A1 (T1063P,H1064W) 49 A1 (T1063P,H1064Y) The parentheses in Table 1 indicate the positions at which amino acid mutations occurred in wild-type botulinum toxin and information on the mutated amino acids. For example, SEQ ID NO: 1 refers to a recombinant polypeptide in which asparagine at position 880 of the receptor-binding domain of wild-type botulinum toxin type A1 is substituted with serine. Example 2: ELISA-based botulinum toxin heavy chain competition assay (ELISA-based BoNT Hc competition assay) Purified recombinant polypeptides of SEQ ID NOs: 2, 6, 33, 34, 35 to 43, and 45 to 49 were analyzed through a competition assay. Differentiated Ntera-2 cells (CRL-1973, ATCC) were seeded in a 96-well plate and cultured for 4 days at 37 °C and 5% CO2 in DMEM / F12 (WelGene) supplemented with 5% FBS (Gibco), B27 (Gibco), and Antibiotic-antimycotic (Gibco). After 4 days, the cells were simultaneously treated with 300 U / well BoNT / A (150 kDa, Medytox, Batch No.: RBTS2301) and BoNT HC (botulinum toxin heavy chain) at varying concentrations for 48 hours. The treatment concentrations of BoNT / A1 WT HC (botulinum toxin type A1 wildtype heavy chain, hereinafter referred to as A1 WT HC) and the recombinant polypeptides were applied at 5-fold intervals, ranging from 5 times up to 15,625 times the molar concentration of BoNT / A (herein, BoNT / A is defined as encompassing both wild-type botulinum toxin serotype A and recombinant botulinum toxin serotype A. It will be apparent to those of ordinary skill in the art that BoNT / A used in the Examples herein refers to wild-type BoNT / A unless otherwise indicated). After treatment with BoNT / A and HC, the medium was replaced with fresh medium, and the cells were further cultured for 2 days, followed by treatment with lysis buffer to perform an ELISA assay. In the ELISA assay, an anti-SNAP25197 antibody (Medytox) was coated onto an immunoplate, and the cell lysate was reacted to capture BoNT / A-cleaved SNAP25, which was then detected using a biotinylated anti-SNAP25 antibody (LS Bio) and streptavidin-HRP (Thermo Scientific). A chromogenic substrate (TMB substrate, Sigma-Aldrich) was added, and the absorbance was measured at a wavelength of 450 nm using a microplate reader (SpectraMax i3, Molecular Devices). GraphPad Prism 7.05 (GraphPad Software Inc., CA, USA) was used for graph presentation and analysis. The half-maximal inhibitory concentration (IC50) was calculated for responses according to HC treatment concentration using a 4-parameter logistic (4PL) non-linear regression method, and the IC50 (uM) value of A1 WT HC was divided by the calculated IC50 (uM) value of the variant HC to determine the relative inhibitory activity compared to wild-type type A1 botulinum toxin heavy chain, as shown in the table below. SEQ ID NO. Relative Inhibitory Activity Compared to Wild Type (IC50 of BoNT A1 HC / IC50 of Recombinant HC) 2 1.2 6 1.4 33 7.4 34 4.0 35 2.4 36 2.9 37 1.8 38 2.2 39 1.3 40 3.8 41 2.1 42 1.5 45 6.2 46 5.6 47 6.8 48 14.4 49 5.5 As a result of the ELISA-based botulinum toxin heavy chain competition assay, the recombinant polypeptide according to an embodiment exhibited a higher relative inhibitory activity than the wild-type receptor-binding domain, confirming enhanced binding affinity for the receptor. Through this, it was confirmed that the recombinant polypeptide of an embodiment may exhibit superior potency and improved safety compared to the wild type. Example 3:  Preparation and Purification of Domain-Substituted Recombinant Botulinum Toxin After confirming the effects of the receptor-binding domains of SEQ ID NOs: 1 to 7 in Examples 1 and 2, recombinant receptor-binding domains further including additional amino acid substitutions in the sequences of SEQ ID NOs: 1 to 7 were designed. For example, the receptor-binding domain included in the recombinant botulinum toxin of SEQ ID NO: 8 includes all amino acid mutations of the recombinant polypeptide of SEQ ID NO: 7, as well as additional mutations. Here, the additional mutations were selected at positions where the amino acids are conserved in wild-type botulinum toxin types A2 and A6 but differ at the corresponding positions of botulinum toxin type A1. At the relevant positions, the amino acids of botulinum toxin type A1 were substituted with amino acids conserved in botulinum toxin types A2 and A6. Subsequently, a recombinant botulinum toxin including the designed recombinant receptor-binding domain was prepared. Specifically, a recombinant full-length botulinum toxin gene including recombinant botulinum toxin type A light chain and heavy chain was cloned by a site-directed mutagenesis method (Site-Directed Mutagenesis) using the pMTL80000 vector system. The cloned constructs are shown in Table 2 below. The recombinant botulinum toxin was produced using a detoxified Hall A-hyper strain in which the toxin gene had been inactivated using the ClosTron system. [Table 2] SEQ ID NO. Light chain Translocation domain Receptor-binding domain 8 A1 A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 9 A1 A1(L681I / D696N / I713T / K 730E / E734K / K779S / N789 D / G804A / E809K / L815V / K 816R / A818V) A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 10 A1-1, A1-2, A4-3, A4-4 A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 11 A1-1, A1-2, A4-3, A4-4 (L260F, I264R) A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 12 A1-1, A1-2, A4-3, A4-4, (L260F, I264R) A1(L681I / D696N / I713T / K 730E / E734K / K779S / N789 D / G804A / E809K / L815V / K 816R / A818V) A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 13 A1-1, A1-2, A4-3, A4-4 A1(L681I / D696N / I713T / K 730E / E734K / K779S / N789 D / G804A / E809K / L815V / K 816R / A818V) A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 14 A1-1, A1-2, A4-3, A4-4, (L260F, I264R) A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, Y1117F, T1232R, L1278F) 15 A1-1, A1-2, A4-3, A4-4, (L260F, I264R) A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R T1232R, L1278F) 16 A1-1, A1-2, A4- 3, A4-4 A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, Y1117F, T1232R, L1278F) 17 A1-1, A1-2, A4-3, A4-4 A1 A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R T1232R, L1278F), 18 A1-1, A1-2, A4- 3, A4-4 A1(D589K, E734K, E809K) A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 19 A1-1, A1-2, A4-3, A4-4(L260F, I264R) A1(D589K, E734K, E809K) A1(N880S, W1068M, N1090S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1232R, L1278F) 22 A1-1, A1-2, A4-3, A4-4 A1 A1(Y1117F, R1273K, L1278F) 23 A1-1, A1-2, A4-3, A4-4 A1 A1(T1232R, R1273K, L1278F) 24 A1-1, A1-2, A4-3, A4-4 A1 A1(N954S) 25 A1-1, A1-2, A4- 3, A4-4 A1 A1(L1278F) 26 A1-1, A1-2, A4-3, A4-4 A1 A1(T1232R) 27 A1-1, A1-2, A4- 3, A4-4 A1 A1(N1025T) 28 A1-1, A1-2, A4-3, A4-4 A1 A1(N1026K) 29 A1-1, A1-2, A4-3, A4-4 A1 A1(Y1117F) 30 A1-1, A1-2, A4-3, A4-4 A1 A1(W1068M, N1090S) 31 A1-1, A1-2, A4-3, A4-4(L260F, I264R) A1 A1(Y1117F, R1273K, L1278F) 32 A1-1, A1-2, A4-3, A4-4(L260F, I264R) A1 A1(T1232R, R1273K, L1278F) 43 A1-1, A1-2, A4- 3, A4-4 A1 R1273K 44 A1 A1(L681I / D696N / I713T / K 730E / E734K / K779S / N789 D / G804A / E809K / L815V / K 816R / A818V) A1 The detoxified Clostridium botulinum strain including a strain in which the toxin gene has been inactivated or knocked out may be used in a method for producing purified recombinant botulinum toxin or a purified recombinant botulinum toxin complex. A method for producing a strain in which the toxin gene has been inactivated or knocked out may employ methods commonly used in the art. The recombinant botulinum toxins of SEQ ID NOs: 8 to 21 produced by the ClosTron method were purified in a 150 kDa form that does not contain complex components. In addition, two recombinant polypeptides (SEQ ID NOs: 20 and 21, respectively) were further prepared by additionally linking IMTP8, a cellpenetrating peptide, to each of the recombinant botulinum toxins of SEQ ID NOs: 10 and 11 using a GS linker. All production of the recombinant botulinum toxins was conducted in a facility licensed for toxin production in accordance with regulatory requirements. Example 4: Evaluation of Thermal Stability of Recombinant Botulinum Toxin Among the recombinant botulinum toxins prepared in Example 3, the thermal stability of the recombinant toxins of SEQ ID NOs: 10 and 11 was compared. The midpoint temperature of thermal denaturation (Tm) was evaluated as a stability assessment method, and in general, a higher midpoint temperature of thermal denaturation is desirable. The thermal stability of each domain-substituted recombinant botulinum toxin type A was assessed using protein thermal shift (PTS) analysis. A protein thermal shift assay was performed according to the manufacturer's instructions using a protein thermal shift dye kit (Catalog No. 4461146, Applied Biosystems). Specifically, 10 pg of each recombinant botulinum toxin was mixed with protein thermal shift dye and buffer to a total volume of 20 pL, after which the ROX signal was monitored using an RT-PCR instrument (C1000 thermal cycler equipped with a CFX96 optical reaction module, Bio-Rad) while the temperature was increased from 20 °C to 95 °C in increments of 1 °C every 60 seconds. Based on the melting curves, the melting temperature (Tm) of each recombinant botulinum toxin was determined. The melting temperatures of each of the recombinant botulinum toxins are shown in FIG. 1 and Table 3. [Table 3] Construct Tm ( °C) △Tm Tm_std Wild type (7s) 52.8 0.0 0.3 SEQ ID NO: 10 (7s) 54.2 1.3 0.3 SEQ ID NO: 11 (7s) 54.3 1.5 0.3 Wild type (7s) 52.7 0.0 0.3 SEQ ID NO: 12 (7s) 55.5 2.8 0.0 SEQ ID NO: 13 (7s) 55.5 2.8 0.0 SEQ ID NO: 14 (7s) 54.5 1.8 0.0 SEQ ID NO: 16 (7s) 54.0 1.3 0.0 SEQ ID NO: 20 (7 s) 56.0 3.3 0.0 The results confirmed that the thermal stability of the recombinant botulinum toxins of SEQ ID NOs: 10 and 11, and SEQ ID NOs: 12, 13, 14, 16, and 20, was increased compared to the wild type. Specifically, whereas the melting point of wild-type botulinum toxin type A1 was 52.8 °C, the melting points of SEQ ID NOs: 10 and 11 increased to 54.2 °C and 54.3 °C, respectively. Whereas the melting point of wild-type botulinum toxin type A1 was 52.7 °C, the melting points of SEQ ID NOs: 12, 13, 14, 16, and 20 increased to 55.5 °C, 55.5 °C, 54.5 °C, 54.0 °C, and 56.0 °C, respectively. Through this, it was confirmed that the thermal stability of the recombinant botulinum toxin according to the present disclosure was significantly improved compared to the wild type. Example 5: Evaluation of the efficacy of recombinant botulinum toxin in mice (1) Recombinant botulinum toxin of SEQ ID NO: 10 Materials and Methods Animals: Female 5-6-week-old CD1 (ICR) mice were purchased from Orient Bio Inc. and used in the experiment at 6-7 weeks of age after 1 week of acclimation and quarantine. Sterilized solid feed for laboratory animals (R40-10, SAFE, France) was provided ad libitum, and tap water was autoclaved and provided ad libitum. During the acclimation, quarantine, and experimental periods, the mice were housed under specific pathogen-free conditions set at a temperature of 23 ± 3 °C, relative humidity of 55 ± 15%, 12-hour light cycle (8:00 AM to 8:00 PM), ventilation rate of 15 times / hour, and illumination of 150-300 Lux. This study was conducted after review and approval by the Medytox Inc. Institutional Animal Care and Use Committee (A-2022-007, A-2024-005). Test substance: A botulinum toxin type A product (BoNT / A), 100 units Coretox (Lot No.: E623001, Medytox), was used. The recombinant botulinum toxin of SEQ ID NO: 10 was prepared at 163 U / mL and used in the experiment. The potency of the recombinant botulinum toxin was measured by a mouse potency assay before the efficacy test, and the test was conducted based on the derived potency value. In the efficacy test, the administration doses of each test substance were equally administered based on units (U: a value representing the biological activity of botulinum toxin, wherein 1 U is the median lethal dose in mice by intraperitoneal administration). The botulinum toxin type A product and the recombinant botulinum toxin of the Example were each diluted to concentrations of 6, 20, and 60 U / mL before administration. The day on which the test substances were administered was designated as Day 0, and after anesthetizing the mice using an injectable anesthetic (100 mg / kg ketamine hydrochloride + 10 mg / kg xylazine), each test substance was administered into the right gastrocnemius muscle of the mice at 0.2 mL / kg using a Hamilton syringe according to the group composition shown in Table 4. [Table 4] Grou P Number of animals Administration substance Dose (U / kg) Route of administration Evaluation parameter 1 6 BoNT / A 1.2 Right gastrocnemius muscle DAS 2 6 BoNT / A 4 Right gastrocnemius muscle DAS 3 6 BoNT / A 12 Right gastrocnemius muscle DAS, body weight 4 6 Recombinant botulinum toxin 1.2 Right gastrocnemius muscle DAS 5 6 Recombinant botulinum toxin 4 Right gastrocnemius muscle DAS 6 6 Recombinant Botulinum Toxin 12 Right gastrocnemius muscle DAS, body weight For the evaluation, the digit abduction score (DAS) method, which visually assesses the degree of muscle paralysis in mice, was used. The DAS values shown in Table 5 (mouse DAS evaluation criteria) indicate the degree of muscle paralysis based on the toe posture on the administered side in mice. [Table 5] Score Criteria 0 Normal; no difference from the shape of the uninjected foot 1 The spaces between the toes are narrowed, or two toes are in contact with each other while the remaining toes are fully extended 2 The spaces between all toes are considerably narrowed, or three toes are in contact with each other 3 The foot is bent and four toes are in contact with each other 4 The foot is bent and all toes are in contact with each other According to the composition of the test groups in Table 4, DAS was assessed from Day 1 after administration until the DAS score reached 0 at administered doses of 1.2, 4, and 12 U / kg. The time at which the DAS score reached 0 for each individual was measured to calculate the mean DAS recovery period. To compare effects on safety in terms of body weight, in the high-dose 12 U / kg administration group, the change in body weight at Week 1 after administration relative to pre-administration body weight was compared between the botulinum toxin type A product-treated group and the recombinant botulinum toxin-treated group. Statistics: GraphPad Prism 7.05 (GraphPad Software Inc., CA, USA) was used for graphical presentation and calculation of DAS ED50, while SPSS software 25.0 (SPSS Inc., IL, USA) and Excel (2013, MS, USA) were used for statistical analysis. A normality test was performed using the Kolmogorov-Smirnov test. Nonparametric data were statistically analyzed using the Mann-Whitney test, whereas parametric data were statistically analyzed using a two-tailed t-test, and a p-value of less than 0.05 was determined to indicate statistical significance. Results: Referring to FIG. 2, following a single administration of the botulinum toxin type A product (BoNT / A: Coretox injection) and the recombinant botulinum toxin of the present application into the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation revealed that significantly increased DAS scores compared to BoNT / A were observed on Days 3, 4, and 7 after administration in the 1.2 U / kg recombinant botulinum toxin-treated group, and on Days 7, 14, and 21 after administration in the 4 U / kg recombinant botulinum toxin-treated group. In the BoNT / A-treated groups at 1.2, 4, and 12 U / kg, recovery of all individuals was observed at Weeks 2, 4, and 6 after administration, respectively, whereas in the recombinant botulinum toxin-treated groups at the same doses, recovery of all individuals was observed at the longer time points of Weeks 3, 5, and 8, respectively (FIG. 2). As a result of calculating the DAS recovery time for each individual and comparing the duration of action, the mean duration of action was confirmed to be 9.0, 19.8, and 33.8 days in the BoNT / A-treated groups at 1.2, 4, and 12 U / kg, respectively, and 17.5, 29.2, and 42.8 days in the recombinant botulinum toxin-treated groups at 1.2, 4, and 12 U / kg, respectively, demonstrating increases in duration of action of 1.9-fold, 1.5-fold, and 1.3-fold at each dose. The duration of action was significantly increased in the 1.2 and 4 U / kg recombinant botulinum toxin-treated groups compared to the BoNT / A-treated groups (Table 6). In the 12 U / kg administration group, the change in body weight at Week 1 relative to pre-administration body weight was 97% on average in the BoNT / A-treated group and 95% on average in the SEQ ID NO: 10-treated group, with no significant difference observed. [Table 6] Mean DAS duration of action (days) Substance 1.2 U / kg 4 U / kg 12 U / kg BoNT / A 9.0 19.8 33.8 SEQ ID NO: 10 17.5** 29.2** 42.8^ Increase rate (fold) 1.9 1.5 1.3 **p<0.01, fp=0.081, compared with the group administered the same dose of BoNT / A, two-tailed t-test or Mann-Whitney test. From the above results obtained in mice, the strong potency and increased duration of action of the recombinant botulinum toxin of the present disclosure compared with botulinum toxin type A product (BoNT / A) were confirmed. (2) Recombinant botulinum toxin of SEQ ID NO: 11 Efficacy evaluation was conducted by the same method as in the example for the recombinant botulinum toxin of SEQ ID NO: 10, except that the recombinant botulinum toxin of SEQ ID NO: 11 was prepared at 384 U / mL and used in the experiment. Results: After a single administration of the botulinum toxin type A product and the recombinant botulinum toxin of SEQ ID NO: 11 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation showed an increase in DAS scores in the 1.2 and 4 U / kg recombinant botulinum toxin of SEQ ID NO: 11 administration groups compared with BoNT / A (FIG. 3). In the BoNT / A administration groups at 1.2, 4, and 12 U / kg, recovery of all animals was observed at Weeks 2, 4, and 6 after administration, respectively, whereas in the recombinant botulinum toxin administration groups at the same doses, recovery of all animals was observed at the extended time points of Weeks 3, 5, and 7, respectively (FIG. 3). As a result of calculating the DAS recovery time for each animal and comparing the duration of action, the mean durations of action in the BoNT / A administration groups at 1.2, 4, and 12 U / kg were confirmed to be 7.2, 16.7, and 31.5 days, respectively, and those in the recombinant botulinum toxin administration groups at 1.2, 4, and 12 U / kg were 14.0, 25.3, and 36.2 days, respectively, showing increases in duration of action of 1.9-fold, 1.5-fold, and 1.1-fold at each dose, respectively; the duration of action was significantly increased in the 1.2 U / kg recombinant botulinum toxin administration group compared with the BoNT / A administration group (Table 7). In the 12 U / kg administration groups, with respect to the change in body weight at Week 1 compared with before administration, the mean was 99% in the BoNT / A administration group and 104% in the SEQ ID NO: 11 administration group, with no decrease in body weight observed. [Table 7] Mean DAS Duration of Action (Days) DAS ED50 (U / kg) Substance 1.2 U / kg 4 U / kg 12 U / kg BoNT / A 7.2 16.7 31.5 2.9 SEQ ID NO: 11 14.0* 25.3^ 36.2 1.9 Increase (fold) 1.9 1.5 1.1 1.5 *p<0.05, fp=0.089, compared with the group administered the same dose of BoNT / A, two-tailed t-test or Mann-Whitney test. From the above results obtained in mice, the strong potency and increased duration of action of the recombinant botulinum toxin of SEQ ID NO: 11 were confirmed compared with botulinum toxin type A product (BoNT / A). Example 6: Evaluation of the efficacy of recombinant botulinum toxin in rats Materials and Methods Animals: Female 6-week-old Sprague-Dawley (SD) rats were purchased from Orient Bio Inc., acclimated and quarantined for 1 week, and 7-week-old rats were used in the experiment. The housing environment was the same as in Example 5. This study was conducted after review and approval by the Medytox Inc. Institutional Animal Care and Use Committee (A-2023-001). Test substance: A botulinum toxin type A product (BoNT / A), 100 units Coretox (Lot No.: E623001, Medytox), was used. The recombinant botulinum toxin of SEQ ID NO: 10 was prepared at 444 U / mL, and the recombinant botulinum toxin of SEQ ID NO: 11 was prepared at 366 U / mL and used in the experiment. For the placebo (vehicle control group), only the botulinum toxin was excluded from the recombinant botulinum toxin, while the remaining excipient components were used identically. The botulinum toxin type A product and recombinant botulinum toxins were each diluted to a concentration of 120 U / mL before administration. The day on which the test substances were administered was designated as Day 0, and after anesthetizing the mice using an injectable anesthetic (60 mg / kg ketamine hydrochloride + 10 mg / kg xylazine), each test substance was administered into the right gastrocnemius muscle of the rats at 0.1 mL / kg using a Hamilton syringe according to the group composition shown in Table 8. [Table 8] G ro u P Numb er of animal s Administered substance Dose (U / kg) Route of administration Evaluation parameters 1 6 Placebo 0 Right gastrocnemius muscle DAS, CMAP 2 6 BoNT / A 12 Right gastrocnemius muscle DAS, CMAP, body weight 3 6 Recombinant botulinum toxin (SEQ ID NO: 10) 12 Right gastrocnemius muscle DAS, CMAP, Body Weight 4 6 Recombinant botulinum toxin (SEQ ID NO: 11) 12 Right gastrocnemius muscle DAS, CMAP, Body Weight For the evaluation, the digit abduction score (DAS) method, which visually evaluates the degree of muscle paralysis in rats, and the compound muscle action potential (CMAP) method, which measures the action potential of muscles in response to external electrical stimulation, were used. The DAS values shown in Table 9 (rat DAS evaluation criteria) indicate the degree of muscle paralysis based on the toe posture on the administered side in rats, and the CMAP values indicate the degree of direct inhibition of muscle contraction resulting from nerve blockade. [Table 9] Score Criteria 0 Normal 1 Loss of abduction of the fifth toe 2 Three toes are in contact with each other 3 Four toes are in contact with each other 4 Loss of abduction of all toes, with the toes in contact with each other CMAP was measured in the left (non-toxin-administered) gastrocnemius muscle area of each rat using an UltraPro S100 device. After anesthesia with an injectable anesthetic, the hair at the measurement site was removed, and the rat was positioned prone. The cathode was placed at the sciatic nerve area on the leg to be measured, and the anode was placed approximately 0.5 cm from that site with reference to the spine. The recording electrode and reference electrode were placed at the gastrocnemius muscle area and Achilles tendon area, respectively, and the ground electrode was placed at the rectus femoris area. The stimulation intensity and duration were set to 25-30 mA and 0.2 ms, respectively, and the amplifier filter range was set to 2-10 K at 60 Hz. Under these conditions, the height from the baseline to the peak of the waveform was recorded as the CMAP measurement value. According to the test group composition in Table 8, DAS was evaluated from Day 3 until the DAS score reached 0. The time at which the DAS score reached 0 for each individual was measured, and the mean DAS recovery period was calculated. CMAP was measured in the left gastrocnemius muscle 7 days after administration to evaluate diffusion to the non-toxin-administered site. In addition, for CMAP measurement, measurements were performed at intervals of 2-3 weeks until recovery of the CMAP value was observed for comparison of CMAPaac (CMAPbaselineauc - CMAPauc), and the values were calculated based on published literature (Dermatologic Surgery 2020; 46(12): e132). Statistics: Same as Example 1 Results: Following a single administration of botulinum toxin type A product and recombinant botulinum toxins to the right gastrocnemius muscle of rats at a dose of 12 U / kg, DAS evaluation showed a significant increase in DAS scores on Days 49 and 56 after administration in the group administered recombinant botulinum toxin of SEQ ID NO: 10 at 12 U / kg, compared to BoNT / A (FIG. 4). An increase in DAS scores compared to BoNT / A was also observed in the group administered recombinant botulinum toxin of SEQ ID NO: 11 (FIG. 4). In the BoNT / A-administered group, recovery of all individuals was observed at Week 10 after administration; in the group administered recombinant botulinum toxin of SEQ ID NO: 10 at the same dose, recovery of all individuals was observed at the increased time point of Week 14; and in the group administered botulinum toxin of SEQ ID NO: 11, recovery of all individuals was observed at Week 11 (FIG. 4). As a result of comparing the duration of action by calculating the DAS recovery time for each individual, the mean duration of action was confirmed to be 50.2 days in the BoNT / A-administered group, while the recombinant botulinum toxin of SEQ ID NO: 10-administered group and the recombinant botulinum toxin of SEQ ID NO: 11-administered group showed significant increases in duration of action of 1.4-fold and 1.3-fold, respectively, at 72.3 days and 65.3 days (Table 10). [Table 10] Mean DAS duration of action (days) Increase rate (fold) Substance 12 U / kg BoNT / A 50.2 n / a SEQ ID NO: 10 72.3* 1.4 SEQ ID NO: 11 65.3* 1.3 *p<0.05, compared with the BoNT / A-treated group, two-tailed t-test. On Day 7 after botulinum toxin administration, CMAP was measured in the gastrocnemius muscle contralateral to the administration site. No significant difference was observed between the recombinant botulinum toxin-treated group of SEQ ID NO: 10 and the BoNT / A-treated group, whereas a significantly increased CMAP value was observed in the recombinant botulinum toxin-treated group of SEQ ID NO: 11 (FIG. 4). As a result of calculating and comparing CMAPaac, the BoNT / A-treated group had a mean of 3558.5, and the botulinum toxin-treated group of SEQ ID NO: 10 had a mean of 3883.3, showing a significant 1.1-fold increase compared with the BoNT / A-treated group, while the botulinum toxin-treated group of SEQ ID NO: 11 had a mean of 3592.5, showing an equivalent or higher value. In the 12 U / kg-treated groups, with respect to body weight change at Week 1 compared with before administration, the botulinum toxin type A product-treated group showed a mean of 107%, the botulinum toxin-treated group of SEQ ID NO: 10 showed a mean of 108%, and the botulinum toxin-treated group of SEQ ID NO: 11 showed a mean of 108%, demonstrating effects equivalent to those of the BoNT / A-treated group. Based on the above results obtained in rats, the strong potency and increased duration of action of the recombinant botulinum toxins compared with the botulinum toxin type A product (BoNT / A) were confirmed, and, in the case of the recombinant botulinum toxin of SEQ ID NO: 11, reduced diffusion compared with the BoNT / A-treated group was confirmed. Example 7: DAS ED50 and IM LD50 Tests of Recombinant Botulinum Toxins in Mice Materials and Methods The animals and housing environment were the same as in Example 5, and this study was conducted after review and approval by the Medytox Inc. Institutional Animal Care and Use Committee (A-2022-007, A-2022-008). Test substance: As the botulinum toxin type A product (BoNT / A), 100 units of Coretox (Lot No.: E623001, Medytox) was used. The recombinant botulinum toxin of SEQ ID NO: 10 was prepared at 1306 U / mL, and the recombinant botulinum toxin of SEQ ID NO: 11 was prepared at 1056 U / mL and used in the experiment. The placebo (vehicle control group) used the same excipient components as the recombinant botulinum toxin, except that the botulinum toxin was excluded. The botulinum toxin type A product and recombinant botulinum toxin were each diluted to concentrations of 6, 20, 60, and 200 U / mL prior to DAS ED50 test administration, and to concentrations of 300, 450, and 675 U / mL prior to IM LD50 test administration. A total of two repeat tests were performed. The day on which the test substances were administered was designated as Day 0. After anesthetizing the mice using an injectable anesthetic (100 mg / kg ketamine hydrochloride + 10 mg / kg xylazine), each test substance was administered at 0.2 mL / kg into the right gastrocnemius muscle of the mice using a Hamilton syringe according to the group allocation (DAS ED50 test groups in Table 11 and IM LD50 test groups in Table 12). [Table 11] G ro u P Numb er of animal s Administered substance Dose (U / kg) Route of administration Evaluation parameter 1 6 Placebo 0 Right gastrocnemius muscle DAS 2 6 BoNT / A 1.2 Right gastrocnemius muscle DAS 3 6 BoNT / A 4 Right gastrocnemius muscle DAS 4 6 BoNT / A 12 Right gastrocnemius muscle DAS 5 6 BoNT / A 40 Right gastrocnemius muscle DAS 6 6 SEQ ID NO: 10 1.2 Right gastrocnemius muscle DAS 7 6 SEQ ID NO: 10 4 Right gastrocnemius muscle DAS 8 6 SEQ ID NO: 10 12 Right gastrocnemius muscle DAS 9 6 SEQ ID NO: 10 40 Right gastrocnemius muscle DAS 1 0 6 SEQ ID NO: 11 1.2 Right gastrocnemius muscle DAS 1 1 6 SEQ ID NO: 11 4 Right gastrocnemius muscle DAS 1 2 6 SEQ ID NO: 11 12 Right gastrocnemius muscle DAS 1 3 6 SEQ ID NO: 11 40 Right gastrocnemius muscle DAS [Table 12] G Numb ro er of Test substance Dose Route of Evaluation u animal (U / kg) administration parameter P s 1 10 BoNT / A 60 Right gastrocnemius muscle Survival 2 10 BoNT / A 90 Right gastrocnemius muscle Survival 3 10 BoNT / A 135 Right gastrocnemius muscle Survival 4 10 SEQ ID NO: 10 60 Right gastrocnemius muscle Survival 5 10 SEQ ID NO: 10 90 Right gastrocnemius muscle Survival 6 10 SEQ ID NO: 10 135 Right gastrocnemius muscle Survival 7 10 SEQ ID NO: 11 60 Right gastrocnemius muscle Survival 8 10 SEQ ID NO: 11 90 Right gastrocnemius muscle Survival 9 10 SEQ ID NO: 11 135 Right gastrocnemius muscle Survival Evaluation was performed using the DAS method, which visually assesses the degree of muscle paralysis in mice. The DAS values indicated in Table 11 represent the degree of muscle paralysis based on toe morphology on the administered side in mice. According to the test group composition in Table 11, DAS was evaluated from Day 1 to Day 14 after administration at administered doses of 1.2, 4, 12, and 40 U / kg, and according to the test group composition in Table 12, the occurrence of deaths was monitored for 14 days after administration at each administered dose. Statistics: In the DAS test, DAS ED50 was calculated for the maximum DAS score of each individual using a 3-parameter logistic model after fixing the minimum value at 0 and the maximum value at 4 using GraphPad Prism. IM LD50 values were calculated by probit analysis using SPSS software (SPSS Inc., IL, USA) for deaths at each administered dose. Results: As shown in Table 11, following a single administration of a botulinum toxin type A product and the recombinant botulinum toxins of the Examples at each dose into the right gastrocnemius muscle of mice, DAS ED50 was calculated. As a result, the mean of two repeated tests was 1.7 U / kg for the BoNT / A administration group, 1.2 U / kg for the recombinant botulinum toxin administration group of SEQ ID NO: 10, and 1.9 U / kg for the recombinant botulinum toxin administration group of SEQ ID NO: 11. An approximately 1.4-fold increase was observed in the recombinant botulinum toxin administration group of SEQ ID NO: 10 compared with the BoNT / A administration group (Table 13). As shown in Table 12, following administration of a botulinum toxin type A product and the recombinant botulinum toxins of the Examples into the right gastrocnemius muscle of mice, the occurrence of deaths was observed for 14 days to calculate the median lethal dose (IM LD50). As a result, the mean of two tests was observed to be 61.7 U / kg for the BoNT / A administration group, 78.0 U / kg for the recombinant botulinum toxin administration group of SEQ ID NO: 10, and 97.1 U / kg for the recombinant botulinum toxin administration group of SEQ ID NO: 11, indicating approximately 1.3-fold and approximately 1.6-fold increases, respectively, for the recombinant botulinum toxin administration groups of SEQ ID NO: 10 and SEQ ID NO: 11 compared with the BoNT / A administration group (Table 13). [Table 13] Article Mean DAS ED50 (U / kg) Mean IM LD50 (U / kg) Safety margin* BoNT / A 1.7 61.7 36.3 SEQ ID NO: 10 1.2 78.0 65.0 SEQ ID NO: 11 1.9 97.1 51.1 *Safety margin: IM LD50 (U / kg) / DAS ED50 (U / kg) As a result of calculating the safety margin from the DAS ED50 and IM LD50 results, the BoNT / A-administered group showed 36.3, the group administered recombinant botulinum toxin of SEQ ID NO: 10 showed 65.0, and the group administered recombinant botulinum toxin 1 of SEQ ID NO: 11 showed 51.1. Compared with the BoNT / A-administered group, increases of 1.8-fold in the group administered recombinant botulinum toxin of SEQ ID NO: 10 and 1.4-fold in the group administered recombinant botulinum toxin of SEQ ID NO: 11 were observed. These results confirmed the strong potency and improved safety of the recombinant botulinum toxin. Example 8: Evaluation of the Efficacy of Recombinant Botulinum Toxin in Mice (1) Recombinant Botulinum Toxins of SEQ ID NOs: 22 and 23 The recombinant botulinum toxin of SEQ ID NO: 22 was prepared at 416 U / mL, and the recombinant botulinum toxin of SEQ ID NO: 23 was prepared at 424 U / mL for the experiment, and the composition of the test groups was as shown in Table 14 below. The test substances or measurement methods used for efficacy evaluation were carried out in the same manner as in the above-described example of the recombinant botulinum toxin of SEQ ID NO: 10. For CMAP measurement, the CMAPaac value was calculated in the same manner as in Example 6. [Table 14] G ro u P Numb er of animal s Administered substance Dose (U / kg ) Route of administration Evaluation parameter 1 6 Placebo Right gastrocnemius muscle DAS, CMAP 2 6 BoNT / A 1.2 Right gastrocnemius muscle DAS 3 6 BoNT / A 4 Right gastrocnemius muscle DAS 4 6 BoNT / A 12 Right gastrocnemius muscle DAS, CMAP, body weight 5 6 SEQ ID NO: 22 1.2 Right gastrocnemius muscle DAS 6 6 SEQ ID NO: 22 4 Right gastrocnemius muscle DAS 7 6 SEQ ID NO: 22 12 Right gastrocnemius muscle DAS, CMAP, body weight 8 6 SEQ ID NO: 23 1.2 Right gastrocnemius muscle DAS 9 6 SEQ ID NO: 23 4 Right gastrocnemius muscle DAS 1 0 6 SEQ ID NO: 23 12 Right gastrocnemius muscle DAS, CMAP, body weight After a single administration of a botulinum toxin type A product and the botulinum toxin of SEQ ID NO: 22 to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation showed that, in the 1.2 U / kg botulinum toxin of SEQ ID NO: 22 administration group, a significant increase in DAS scores compared with BoNT / A was observed on days 1, 3, 4, and 7 after administration. In the botulinum toxin of SEQ ID NO: 23 administration group, an increase in DAS scores compared with BoNT / A was observed in the 1.2 and 4 U / kg administration groups (FIG. 5). As a result of calculating the DAS recovery time for each individual and comparing the duration of action, the BoNT / A 1.2, 4, and 12 U / kg administration groups were confirmed to have mean durations of 9.8, 21.0, and 25.3 days, respectively, while the botulinum toxin of SEQ ID NO: 22 administration groups showed mean durations of 15.2, 23.0, and 37.4 days, respectively. That is, compared with wild-type BoNT / A, the duration of action of the recombinant polypeptide of the present disclosure was observed to increase by 1.6-fold, 1.1-fold, and 1.5-fold at each dose, respectively. The DAS recovery times in the botulinum toxin of SEQ ID NO: 23 administration groups were mean 16.3, 27.3, and 35.5 days, respectively, and increases in duration of action of 1.7-fold, 1.3-fold, and 1.4-fold were observed at each dose, respectively (Table 15). In addition, in the 12 U / kg administration groups of the botulinum toxin of SEQ ID NO: 22 and the botulinum toxin of SEQ ID NO: 23, the duration of action was significantly increased compared with the BoNT / A administration group (Table 15). The DAS ED50 values were determined to be 3.2 U / kg for the BoNT / A administration group, 1.1 U / kg for the SEQ ID NO: 22 administration group, and 2.1 U / kg for the SEQ ID NO: 23 administration group, indicating that potency was 2.9-fold and 1.5-fold greater, respectively, than that of the BoNT / A administration group. As a result of CMAP measurement on day 7 after administration, a significant decrease in CMAP values at the administration site was observed in the botulinum toxin of SEQ ID NO: 22 and botulinum toxin of SEQ ID NO: 23 administration groups compared with the BoNT / A administration group, confirming greater potency, while no significant difference was observed in CMAP values at the contralateral site compared with the BoNT / A administration group (FIG. 6). When the CMAPaac values were calculated and compared, the BoNT / A administration group had a mean of 2077.1, the group administered the botulinum toxin of SEQ ID NO: 22 had a mean of 2680.3, and the group administered the botulinum toxin of SEQ ID NO: 23 had a mean of 2612.9, and a significant 1.3-fold increase was observed in both of these groups compared with the BoNT / A administration group. In the 12 U / kg administration groups, the body weight changes at Week 1 compared with before administration were a mean of 96% in the BoNT / A administration group, a mean of 96% in the botulinum toxin of SEQ ID NO: 22 administration group, and a mean of 97% in the botulinum toxin of SEQ ID NO: 23 administration group, showing effects equivalent to those of the BoNT / A administration group. From the above results obtained in mice, the greater potency and increased duration of action of the recombinant polypeptides, namely the botulinum toxin of SEQ ID NO: 22 and the botulinum toxin of SEQ ID NO: 23, were confirmed compared with botulinum toxin type A product (BoNT / A). [Table 15] Mean DAS duration of action (days) Increase rate (fold) Substance 1.2 U / kg 4 U / kg 12 U / kg 1.2 U / kg 4 U / kg 12 U / kg BoNT / A 9.8 21.0 25.3 N / A N / A N / A SEQ ID NO: 22 15.2b 23.0 37.4* 1.6 1.1 1.5 SEQ ID NO: 23 16.3a 27.3a 35.5* 1.7 1.3 1.4 *p<0.05, ap=0.05, bp=0.08, compared with the group administered the same dose of BoNT / A, two-tailed t-test or Mann-Whitney test (2) Recombinant botulinum toxins of SEQ ID NOs: 14 and 15 The botulinum toxin type A product (BoNT / A) was the same as in Example 1, and the recombinant botulinum toxin of SEQ ID NO: 14 was prepared at 394 U / mL and the recombinant botulinum toxin of SEQ ID NO: 15 was prepared at 419 U / mL for use in the experiment. The composition of the test groups was as shown in Table 16, and the remaining details were the same as those described in (1) of Example 8 above. However, CMAPaac was not calculated. [Table 16] G ro u P Numb er of animal s Test substance Dose (U / kg) Route of administration Evaluation parameter 1 6 Placebo - Right gastrocnemius muscle DAS, CMAP 2 6 BoNT / A 1.2 Right gastrocnemius muscle DAS 3 6 BoNT / A 4 Right gastrocnemius muscle DAS 4 6 BoNT / A 12 Right gastrocnemius muscle DAS, CMAP 5 6 SEQ ID NO: 14 1.2 Right gastrocnemius muscle DAS 6 6 SEQ ID NO: 14 4 Right gastrocnemius muscle DAS 7 6 SEQ ID NO: 14 12 Right gastrocnemius muscle DAS, CMAP 8 6 SEQ ID NO: 15 1.2 Right gastrocnemius muscle DAS 9 6 SEQ ID NO: 15 4 Right gastrocnemius muscle DAS 1 0 6 SEQ ID NO: 15 12 Right gastrocnemius muscle DAS, CMAP After a single administration of a botulinum toxin type A product and recombinant botulinum toxins to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation showed that, in the case of the botulinum toxin of SEQ ID NO: 14, an increase in the DAS score compared to BoNT / A was observed in the 4 U / kg administration group (FIG. 7). Based on comparison of the duration of action by calculating the DAS recovery time for each individual, an increased duration of action equal to or greater than that of BoNT / A was observed in the recombinant botulinum toxin administration groups (Table 17). As a result of calculating DAS ED50, the BoNT / A administration group showed 5.8 U / kg, the SEQ ID NO: 14 administration group showed 3.3 U / kg, and the SEQ ID NO: 15 administration group showed 3.0 U / kg, demonstrating 1.8-fold and 1.9-fold greater potency, respectively, compared to the BoNT / A administration group. As a result of CMAP measurement on Day 7 after administration, a significant decrease in the CMAP value at the administration site was observed in the administration groups of the botulinum toxin of SEQ ID NO: 14 and the botulinum toxin of SEQ ID NO: 15 compared to the BoNT / A administration group, confirming strong potency. In addition, a significant increase in CMAP at the non-administration site was observed in the administration group of the botulinum toxin of SEQ ID NO: 15 compared to the BoNT / A administration group, confirming an effect of inhibiting diffusion (FIG. 8). In the 12 U / kg administration groups, with respect to body weight change at Week 1 compared to before administration, the BoNT / A administration group showed a mean value of 95% and the SEQ ID NO: 14 administration group showed a mean value of 97%, with no significant difference compared to the BoNT / A administration group. The SEQ ID NO: 15 administration group showed a mean value of 111%, demonstrating a significant increase in body weight compared to the BoNT / A administration group (p < 0.001). From the above results obtained in mice, the strong potency, increased duration of action, and improved effect of inhibiting diffusion of the recombinant botulinum toxins of the present application compared to a botulinum toxin type A product (BoNT / A) were confirmed. [Table 17] Mean DAS duration of action (days) Increase rate (fold) Substance 1.2 U / kg 4 U / kg 12 U / kg 1.2 U / kg 4 U / kg 12 U / kg BoNT / A 10.5 20.8 26.3 N / A N / A N / A SEQ ID NO: 14 13.8 20.5 33.8 1.3 1.0 1.3 SEQ ID NO: 15 14.7 22.8 31.5 1.4 1.1 1.2 (3) Recombinant botulinum toxins of SEQ ID NOs: 12 and 13 The recombinant botulinum toxin of SEQ ID NO: 12 was prepared at 559 U / mL, and the recombinant botulinum toxin of SEQ ID NO: 13 was prepared at 440 U / mL, and the composition of the test groups was as shown in Table 18 below. The test substances or measurement methods used for efficacy evaluation were carried out in the same manner as in the above-described examples of the recombinant botulinum toxins of SEQ ID NOs: 22 and 23. [Table 18] G ro u P Number of animals Administered substance Dose (U / kg) Route of administration Evaluation parameter 1 6 Placebo 0 Right gastrocnemius muscle DAS, CMAP 2 6 BoNT / A 1.2 Right gastrocnemius muscle DAS 3 6 BoNT / A 4 Right gastrocnemius muscle DAS 4 6 BoNT / A 12 Right gastrocnemius muscle DAS, CMAP, body weight 5 6 SEQ ID NO: 12 1.2 Right gastrocnemius muscle DAS 6 6 SEQ ID NO: 12 4 Right gastrocnemius muscle DAS 7 6 SEQ ID NO: 12 Right gastrocnemius DAS, CMAP, body 12 muscle weight 8 6 SEQ ID NO: 13 1.2 Right gastrocnemius muscle DAS 9 6 SEQ ID NO: 13 4 Right gastrocnemius muscle DAS 1 6 SEQ ID NO: 12 Right gastrocnemius DAS, CMAP, body 0 13 muscle weight Following a single administration of botulinum toxin type A product and the botulinum toxins of SEQ ID NOs: 12 and 13 to the right gastrocnemius muscles of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation showed increased DAS scores in the 1.2 and 4 U / kg dose groups compared with BoNT / A, demonstrating equivalent or greater effects, while the 12 U / kg dose group showed an equivalent effect (FIG. 9). As a result of calculating the DAS recovery time for each individual and comparing the duration of action, the duration of action was significantly increased in the 1.2 U / kg dose group of the botulinum toxin of SEQ ID NO: 12 and in the 4 U / kg dose group of the botulinum toxin of SEQ ID NO: 13, compared with the BoNT / A-treated group (Table 19). Based on the DAS ED50 calculation results, the BoNT / A-treated group was 3.0 U / kg, the botulinum toxin of SEQ ID NO: 12-treated group was 2.5 U / kg, and the botulinum toxin of SEQ ID NO: 13-treated group was 2.8 U / kg, demonstrating 1.2-fold and 1.1-fold greater potency, respectively, compared with the BoNT / A-treated group. CMAP measurements on day 7 after administration showed equivalent effects on CMAP values at the administration site, while a significant increase in CMAP values at the contralateral site was observed in the SEQ ID NO: 12-treated group (FIG. 10). Comparison based on calculated CMAPaac values showed that the BoNT / A-treated group had a mean value of 2502.2, while the botulinum toxin of SEQ ID NO: 13-treated group had a mean value of 3041.9, demonstrating a significant 1.2-fold increase compared with the BoNT / A-treated group. In the 12 U / kg-treated groups, the body weight change at Week 1 relative to before administration was a mean value of 96% in the BoNT / A-treated group, a mean value of 95% in the botulinum toxin of SEQ ID NO: 12-treated group, and a mean value of 97% in the botulinum toxin of SEQ ID NO: 13-treated group, demonstrating equivalent effects compared with the BoNT / A-treated group. Based on the above results obtained in mice, the strong potency, increased duration of action, and improved effect of inhibiting diffusion of the recombinant botulinum toxins of SEQ ID NOs: 12 and 13 were confirmed compared with the botulinum toxin type A product (BoNT / A). [Table 19] Mean DAS duration of action (days) Increase rate (fold) Substance 1.2 U / kg 4 U / kg 12 U / kg 1.2 U / kg 4 U / kg 12 U / kg BoNT / A 5.5 16.3 38.7 N / A N / A N / A SEQ ID NO: 12 11.2** 21.3 38.0 2.0 1.3 1.0 SEQ ID NO: 13 8.8 25.0* 29.3 1.6 1.5 0.8 *p<0.05, **p<0.01, compared with the group administered the same dose of BoNT / A, two-tailed t-test (4) Recombinant botulinum toxin of SEQ ID NO: 16 The recombinant botulinum toxin of SEQ ID NO: 16 was prepared at 444 U / mL, and the test group composition was as shown in Table 20 below. The test substances or measurement methods used for efficacy evaluation were carried out in the same manner as in the above-described examples of the recombinant botulinum toxins of SEQ ID NOs: 22 and 23. [Table 20] G ro u P Number of animals Administered substance Dose (U / kg) Route of administratio n Evaluation parameter 1 6 Placebo 0 Right gastrocnemiu s muscle DAS, CMAP 2 6 BoNT / A 1.2 Right gastrocnemiu s muscle DAS 3 6 BoNT / A 4 Right gastrocnemiu s muscle DAS 4 6 BoNT / A 12 Right gastrocnemiu s muscle DAS, CMAP, body weight 5 6 SEQ ID NO: 16 1.2 Right gastrocnemiu s muscle DAS 6 6 SEQ ID NO: 16 4 Right gastrocnemiu s muscle DAS 7 6 SEQ ID NO: 16 12 Right gastrocnemiu s muscle DAS, CMAP, body weight Following a single administration of a botulinum toxin type A product and the recombinant botulinum toxin of SEQ ID NO: 16 into the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation showed a significant increase in DAS score compared to BoNT / A on day 21 after administration in the group administered 12 U / kg of the recombinant botulinum toxin of SEQ ID NO: 16, and increased DAS scores compared to BoNT / A were also observed at other measurement time points (FIG. 11). As a result of calculating the DAS recovery time for each individual and comparing the duration of action, the duration of action was significantly increased in the group administered 4 U / kg of the recombinant botulinum toxin of SEQ ID NO: 16 compared to the BoNT / A-administered group (Table 21). As a result of calculating DAS ED50, the BoNT / A-administered group had a value of 2.0 U / kg, while the group administered the recombinant botulinum toxin of SEQ ID NO: 16 had a value of 1.8 U / kg, demonstrating 1.1-fold greater potency compared to the BoNT / A-administered group. As a result of CMAP measurement on Day 7 after administration, a significant decrease in the CMAP value at the administration site was observed in the group administered the recombinant botulinum toxin of SEQ ID NO: 16 compared to the BoNT / A-administered group, confirming greater potency, whereas no significant difference was observed in the CMAP value at the contralateral site compared to the BoNT / A-administered group (FIG. 12). As a result of calculating and comparing CMAPaac, the BoNT / A-administered group had a mean value of 2061.9, while the group administered the botulinum toxin of SEQ ID NO: 16 had a mean value of 2659.7, demonstrating a significant 1.3-fold increase compared to the BoNT / A-administered group. In the 12 U / kg-administered groups, the change in body weight at Week 1 compared to before administration was 94% on average in the BoNT / A-administered group and 93% on average in the group administered the recombinant botulinum toxin of SEQ ID NO: 16, with no significant difference observed compared to the BoNT / A-administered group. Based on the above results obtained in mice, the greater potency and increased duration of action of the recombinant botulinum toxin of SEQ ID NO: 16, a recombinant botulinum toxin, were confirmed compared to the botulinum toxin type A product (BoNT / A). [Table 21] Mean DAS duration of action (days) Increase rate (fold) Substance 1.2 U / kg 4 U / kg 12 U / kg 1.2 U / kg 4 U / kg 12 U / kg BoNT / A 11.2 18.7 32.7 N / A N / A N / A SEQ ID NO: 16 13.8 26.8* 45.5a 1.2 1.4 1.4 *p<0.05, ap=0.05, compared with the group administered the same dose of BoNT / A, two-tailed t-test or Mann-Whitney test Example 9: Evaluation of the efficacy of recombinant botulinum toxins of SEQ ID NOs: 24 to 32 and 43 in mice Efficacy evaluation was conducted under the same conditions for animals and housing environment as in Example 5. Test substance: A botulinum toxin type A product (BoNT / A), 100 units Coretox (Lot No.: E623001, Medytox), was used. The recombinant botulinum toxins were prepared in liquid form and used in the experiment, and were diluted to concentrations of 6, 20, and 60 U / mL before administration. Ten types of recombinant botulinum toxins were used in the experiment, and a total of four tests were performed. The botulinum toxin type A product and recombinant botulinum toxins were administered to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, followed by comparison of the duration of action through DAS evaluation and comparison of body weight at 1 week after administration at a dose of 12 U / kg. Results: Following a single administration of the botulinum toxin type A product (BoNT / A: Coretox injection) and 10 types of recombinant botulinum toxins to the right gastrocnemius muscle of mice at doses of 1.2, 4, and 12 U / kg, DAS evaluation showed that the recombinant botulinum toxin of SEQ ID NO: 24 exhibited 1.6-fold and 1.1-fold increases in DAS duration of action at doses of 1.2 and 4 U / kg, respectively, compared with BoNT / A, while an equivalent duration of action was observed at 12 U / kg. The recombinant botulinum toxin of SEQ ID NO: 25 exhibited 1.3-fold, 1.4-fold, and 1.2-fold increases in DAS duration of action at doses of 1.2, 4, and 12 U / kg, respectively, compared with BoNT / A; the recombinant botulinum toxin of SEQ ID NO: 26 exhibited 1.2-fold, 1.1-fold, and 1.3-fold increases in DAS duration of action at doses of 1.2, 4, and 12 U / kg, respectively, compared with BoNT / A; and the recombinant botulinum toxin of SEQ ID NO: 27 exhibited 1.2-fold, 1.1-fold, and 1.1-fold increases in DAS duration of action at doses of 1.2, 4, and 12 U / kg, respectively, compared with BoNT / A. The recombinant botulinum toxin of SEQ ID NO: 28 exhibited 1.2-fold and 1.3-fold increases in DAS duration of action at doses of 1.2 and 4 U / kg, respectively, compared with BoNT / A, and an equivalent DAS duration of action to BoNT / A was observed at a dose of 12 U / kg. The recombinant botulinum toxins of SEQ ID NOs: 29 to 31 exhibited DAS durations of action equivalent to that of BoNT / A at doses of 1.2, 4, and 12 U / kg. The recombinant botulinum toxin of SEQ ID NO: 32 exhibited 1.6-fold and 1.3-fold increases in DAS duration of action at doses of 1.2 and 4 U / kg, respectively, compared with BoNT / A, and an equivalent DAS duration of action to BoNT / A was observed at a dose of 12 U / kg. SEQ ID NO: 43 exhibited a 1.4-fold increase in DAS duration of action at a dose of 1.2 U / kg compared with BoNT / A, and durations of action equivalent to that of BoNT / A at doses of 4 U / kg and 12 U / kg. In addition, safety was evaluated by monitoring body weight changes after administration of the recombinant botulinum toxins of SEQ ID NOs: 28 to 32 and 43 at doses of 1.2, 4, and 12 U / kg. As a result, all showed body weight loss equivalent to that of wild-type BoNT / A, confirming that they exhibit safety at least equivalent to that of wildtype botulinum toxin. Based on the above results obtained in mice, the recombinant botulinum toxins were confirmed to have increased durations of action or safety equivalent to or greater than that of the wild-type botulinum toxin type A product (BoNT / A). Example 10: Evaluation of the efficacy of recombinant botulinum toxins of SEQ ID NOs: 18 and 19 in mice Efficacy evaluation was conducted under the same conditions for animals and housing environment as in Example 5. Test substance: The botulinum toxin type A product (BoNT / A) used was 100 units of Coretox (Lot No.: E623001, Medytox). The recombinant botulinum toxins were prepared in liquid form and used in the experiment, and were diluted to a concentration of 60 U / mL before administration. Two types of recombinant botulinum toxins were used in the experiment, and a total of two tests were performed. After administering the botulinum toxin type A product and the recombinant botulinum toxins into the right gastrocnemius muscle of mice at a dose of 12 U / kg, potency was compared by compound muscle action potential (CMAP) assessment, and body weights were compared at 1 week after administration. Results: After a single administration of the botulinum toxin type A product (BoNT / A: Coretox injection) and the recombinant botulinum toxin of SEQ ID NO: 18 into the right gastrocnemius muscle of mice at a dose of 12 U / kg, CMAP values at the administration site were compared at 1 week after administration. As a result, a significant decrease was observed in the SEQ ID NO: 18 administration group, with a mean of 0.1 mV, compared with a mean CMAP value of 0.9 mV in the BoNT / A administration group (p<0.001). For body weight change at 1 week, body weight changes of 97% in the BoNT / A administration group and 93% in the SEQ ID NO: 18 administration group were observed, with no significant difference observed. After a single administration of the recombinant botulinum toxin of SEQ ID NO: 19 at a dose of 12 U / kg, CMAP values at the administration site were compared at 1 week after administration. As a result, a significant decrease was observed in the botulinum toxin of SEQ ID NO: 19 administration group, with a mean of 0.3 mV, compared with a mean CMAP value of 1.3 mV in the BoNT / A administration group (p<0.01). For body weight change at Week 1, body weight changes of 93% were observed in both the BoNT / A and botulinum toxin of SEQ ID NO: 19 administration groups, demonstrating equivalent effects. From the above results obtained in mice, the strong potency and equivalent safety of the recombinant botulinum toxins of SEQ ID NOs: 18 and 19, as compared with the botulinum toxin type A product (BoNT / A), were confirmed. Example 11: Cell-Based Activity of Recombinant Botulinum Toxin of SEQ ID NO: 44 Materials and Methods *The cell line used in the experiment was the same as that in Example 2, and the cells were treated for 48 hours with BoNT / A (150 kDa, Medytox, Batch No.: RBTS2301) and the recombinant botulinum toxin of SEQ ID NO: 44 (150 kDa, Medytox), serially diluted to 300, 60, 12, 2.4, and 0.48 U / well. Thereafter, the cell culture medium was replaced, followed by additional culture for 48 hours, after which the cells were treated with lysis buffer to perform an ELISA assay. The method for performing the ELISA assay was the same as that in Example 2. Results As a result of analyzing the activities of cell-based wild-type and recombinant botulinum toxins by the ELISA method, an 11-fold increase in activity relative to the wildtype botulinum toxin was confirmed in the recombinant botulinum toxin-treated group. [Table 22] SEQ ID NO. EC50 (U / well) Relative increase in activity compared with wild type BoNT / A 0.866 - SEQ ID NO: 44 0.078 11-fold increase Through this, it was confirmed that the recombinant polypeptide may exhibit superior potency compared to the wild type. A recombinant polypeptide including a receptor-binding domain having an amino acid mutation according to an embodiment exhibits superior efficacy compared to a wildtype receptor-binding domain. A botulinum toxin including a recombinant polypeptide according to an embodiment exhibits increased potency, safety, or half-life compared to wild-type botulinum toxin.

Claims

1. A recombinant polypeptide comprising a receptor-binding domain of botulinum toxin type A1, wherein the receptor-binding domain comprises an amino acid mutation at one or more positions among positions that are conserved as common amino acids in botulinum toxin type A2 and botulinum toxin type A6 but have different amino acids at corresponding positions in botulinum toxin type A1.

2. The recombinant polypeptide of claim 1, wherein the positions of the amino acid mutation comprise at least one selected from the group consisting of N880, W1068, and N1090.

3. The recombinant polypeptide of claim 1, wherein the receptor-binding domain comprises at least one amino acid mutation selected from the group consisting of N880S, W1068M, and N1090S.

4. The recombinant polypeptide of claim 2, wherein the receptor-binding domain further comprises an amino acid mutation at one or more positions selected from the group consisting of N954, M968, T990, Q991, N1025, N1026, N1052, T1063, H1064, Y1117, T1232, and L1278.

5. The recombinant polypeptide of claim 4, wherein the receptor-binding domain further comprises at least one amino acid mutation selected from the group consisting of N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, Y1117F, T1232R, and L1278F.

6. The recombinant polypeptide of claim 1, further comprising a translocation domain, wherein the translocation domain comprises a polypeptide selected from the group consisting of a translocation domain of botulinum toxin type A1, a translocation domain of botulinum toxin type A2, a translocation domain of botulinum toxin type A6, and variants thereof.

7. The recombinant polypeptide of claim 6, comprising a translocation domain of botulinum toxin type A1, wherein the translocation domain comprises an amino acid mutation at one or more positions that are conserved as common amino acids in botulinum toxin type A2 and botulinum toxin type A6 but have different amino acids at corresponding positions in botulinum toxin type A1; or comprising a translocation domain of botulinum toxin type A1 and comprising an amino acid mutation at position 589.

8. The recombinant polypeptide of claim 6, comprising a translocation domain of botulinum toxin type A1, wherein the positions of amino acid mutations in the translocation domain comprise at least one selected from the group consisting of D589, L681, D696, I713, K730, E734, K779, N789, G804, E809, L815, K816, and A818.

9. The recombinant polypeptide of claim 1, further comprising a light chain domain, wherein the light chain domain comprises a polypeptide selected from the group consisting of botulinum toxin type A1, botulinum toxin type A4, and variants thereof.

10. The recombinant polypeptide of claim 9, wherein the light chain domain comprises a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of botulinum toxin type A4 light chain, or variants thereof; or comprises a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of botulinum toxin type A4 light chain, or variants thereof, wherein leucine at position 260 of the light chain domain is substituted with phenylalanine, and isoleucine at position 264 is substituted with arginine.

11. The recombinant polypeptide of claim 1, wherein the receptor-binding domain has a lower half-maximal inhibitory concentration value according to a botulinum toxin heavy chain competition assay than that of a wild-type receptor-binding domain.

12. The recombinant polypeptide of claim 1, further comprising a light chain domain and a translocation domain, wherein the recombinant polypeptide has increased potency, safety, or half-life compared to wild-type botulinum toxin type A.

13. The recombinant polypeptide of claim 12, wherein the recombinant polypeptide comprises at least one selected from recombinant botulinum toxins of SEQ ID NO: 8 to SEQ ID NO: 32, SEQ ID NO: 43, and SEQ ID NO:

44.

14. A recombinant polypeptide comprising a receptor-binding domain of botulinum toxin type A1, wherein the receptor-binding domain comprises amino acid mutations at two or more positions selected from N954, Y1117, T1232, R1273, and L1278.

15. The recombinant polypeptide of claim 14, wherein the receptor-binding domain comprises two or more amino acid mutations selected from N954S, Y1117F, T1232R, R1273K, and L1278F.

16. The recombinant polypeptide of claim 14, wherein the receptor-binding domain comprises amino acid mutations at two or more positions selected from N954S, T1232R, and L1278F.

17. The recombinant polypeptide of claim 14, wherein the amino acid mutations of the receptor-binding domain comprise N954S and T1232R; N954S and L1278F; T1232R and L1278F; T1232R and R1273K; R1273K and L1278F; N954S, T1232R, and L1278F; Y1117F, R1273K, and L1278F; or T1232R, R1273K, and L1278F.

18. A recombinant polypeptide comprising a variant of a receptor-binding domain of botulinum toxin type A1, wherein the variant of the receptor-binding domain has a lower halfmaximal inhibitory concentration value in a botulinum toxin heavy chain competition assay than that of a wild-type receptor-binding domain.

19. The recombinant polypeptide of claim 18,wherein the variant of the receptor-binding domain comprises an amino acid mutation at one or more positions selected from the group consisting of N880, N954, M968, T990, Q991, N1025, N1026, N1052, T1063, H1064, W1068, N1090, Y1117, S1142, T1232, and L1278.

20. A recombinant botulinum toxin comprising a variant in which one or moreamino acids of a receptor-binding domain sequence of botulinum toxin type A1 are mutated,wherein the amino acid mutations comprise one or more selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, Y1117F,S1142Y, S1142R, S1142I, S1142F, S1142H, T1232R, and L1278F, and the recombinant botulinum toxin, when administered to a subject in need of treatment, is superior in one or more of potency and safety as compared with wild-type botulinum toxin type A1, with the remainder being at least equivalent to that of the wild-type toxin.

21. The recombinant botulinum toxin of claim 20,comprising a light chain domain selected from the group consisting of the botulinum toxin type A1 light chain, botulinum toxin type A4 light chain, and variants thereof.

22. The recombinant botulinum toxin of claim 21,wherein the light chain domaincomprises a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of botulinum toxin type A4 light chain, or variants thereof; orcomprises a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of botulinum toxin type A4 light chain, or variants thereof, wherein leucine at position 260 of the light chain domain is substituted with phenylalanine, and isoleucine at position 264 is substituted with arginine.

23. The recombinant botulinum toxin of claim 20,comprising a translocation domain,wherein the translocation domain comprises an amino acid mutation at one or more positions among positions that are conserved as common amino acids in botulinum toxin type A2 and botulinum toxin type A6 but have different amino acids at corresponding positions in botulinum toxin type A1; or the translocation domain comprises a translocation domain of botulinum toxin type A1 and comprises at least one amino acid mutation selected from the group consisting of D589, E734, and E809.

24. A recombinant botulinum toxin comprising a variant of botulinum toxin type A light chain and a receptor-binding domain comprising one or more amino acid mutations selected from the group consisting of N880S, N954S, M968I, T990N, Q991K, N1025T, N1026K, N1052K, T1063P, H1064R, W1068M, N1090S, S1142Y, S1142R, S1142I, S1142F, S1142H, Y1117F, T1232R, and L1278F.

25. The recombinant botulinum toxin of claim 24, wherein the receptor-binding domain comprises a recombinant polypeptide of SEQ ID NOs: 1 to 7, 33 to 42, and 45 to 49.

26. The recombinant botulinum toxin of claim 24, wherein the variant of botulinum toxin type A light chain comprises: a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of botulinum toxin type A4 light chain, or variants thereof; or a first domain and a second domain of the botulinum toxin type A1 light chain, or variants thereof; and a third domain and a fourth domain of botulinum toxin type A4 light chain, or variants thereof, wherein leucine at position 260 is substituted with phenylalanine and isoleucine at position 264 is substituted with arginine.

27. The recombinant botulinum toxin of claim 24, wherein the recombinant botulinum toxin exhibits a higher value in one or more of safety and duration of action upon administration to a subject, compared to wild-type botulinum toxin type A1.

28. A recombinant polypeptide comprising a botulinum toxin type A1 translocation domain, wherein the translocation domain comprises an amino acid mutation at one or more positions that are conserved as common amino acids in botulinum toxin type A2 and botulinum toxin type A6 but have different amino acids at corresponding positions in botulinum toxin type A1.

29. The recombinant polypeptide of claim 28, wherein the positions of the amino acid mutations in the translocation domain comprise at least one selected from the group consisting of L681, D696, I713, K730, E734, K779, N789, G804, E809, L815, K816, and A818.

30. The recombinant polypeptide of claim 28, wherein the translocation domain comprises at least one amino acid mutation selected from the group consisting of L681I, D696N, I713T, K730E, E734K, K779S, N789D, G804A, E809K, L815V, K816R, and A818V.

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