High concentration botulinum toxin treatment for neck rejuvenation

Abstract

The present invention generally relates to the use of botulinum toxin for neck rejuvenation by injecting a high concentration of botulinum toxin into the latissimus latipes. In particular, a botulinum toxin is injected in at least one latissimus latius belt of a subject at a concentration greater than 100 U / ml. Furthermore, the present invention relates to a method for rejuvenating the neck comprising injecting a high concentration of botulinum toxin.

Description

Technical Field

[0001] The present invention generally relates to the use of botulinum toxin for neck rejuvenation by injecting a high concentration of botulinum toxin into the platysma muscle. Specifically, the botulinum toxin is injected at a concentration greater than 100 U / ml into at least one platysma band of a subject, and optionally into the platysma muscle at the mandibular contour. In addition, the present invention relates to a method for neck rejuvenation, comprising injecting a high concentration of botulinum toxin. Background Art

[0002] Restoring the volume of the mid-face and treating the wrinkles of the upper face have been the focus of facial rejuvenation for many years. However, in the past few years, the lower face and neck have become the focus of facial rejuvenation, especially in the older age groups. A sharp, defined mandibular contour is associated with a youthful appearance, and a slender, streamlined neck is considered a sign of health and youth. However, with aging, the mandibular contour loses its sharpness, resulting in a blurred or sagging contour, and the neck begins to show platysma bands, sagging skin, and prominent folds or jowls.

[0003] The platysma muscle is a thin layer of muscle that wraps around the entire neck. It originates from the fascia covering the deltoid and upper part of the pectoralis muscles. Its fine muscle fibers cross the clavicle and run obliquely upward, laterally, and medially to cover the neck. The platysma muscle passes through the mandible, with some of its fibers inserting into the bone below the oblique line, and other fibers extending to about 4 cm above the mandibular margin, entering the cheek, and inserting into the skin and subcutaneous tissue. In addition, some muscle fibers are embedded in the mentalis muscle and the depressor anguli oris muscle.

[0004] Functionally, the platysma muscle is an expressive muscle that contributes to facial expressions. It expands the skin of the neck and pulls it downward. Viewed from the side, it acts in concert with the depressor anguli oris muscle (DAO) to lower the lower lip angle by pulling down the outer corner of the mouth. It is also associated with the formation of neck wrinkles. Thus, contraction of the platysma muscle pulls down the mandible, the corners of the mouth, and the lower lip. During the aging process, the muscle begins to separate and bunch up into bundles or bands, typically alongside the midline on each side of the neck and on the sides of the neck. These bundles or bands are called platysma bands and are characteristic of an aging neck.

[0005] A variety of surgical techniques for treating the platysma band have been used to improve or restore the contour of the neck. However, adverse surgical outcomes and the understanding that the platysma band is caused by muscle activity during the aging process (see, e.g., Trévidic, P. and Criollo-Lamilla, G., Plast. Reconstr. Surg. 2017; 139(1):41-47) have shifted the treatment towards non-surgical methods such as injecting botulinum toxin. Botulinum toxin is a neurotoxin from Clostridium botulinum that blocks the release of acetylcholine into the synaptic cleft, thereby preventing cholinergic neuromuscular innervation. It exists in different serotypes (types A-H) and is produced as a non-covalent macromolecular protein complex consisting of a 150 kDa active neurotoxin and a non-toxic neurotoxin-associated protein (NAP or non-toxic "complexing protein"). The 150 kDa neurotoxin is synthesized as a single-chain polypeptide that is proteolytically cleaved into two chains, namely a 50 kDa light chain and a 100 kDa heavy chain, which are linked by a disulfide bond.

[0006] Botulinum toxin can relax and flatten the platysma band, making the band less prominent. It can also improve transverse (horizontal) neck wrinkles. In addition, by reducing the degree of downward pull, injection of the platysma muscle can also be used to treat the lower face contour, such as the contour of the mandible, for overall facial rejuvenation. In addition, the reduced downward pull of the platysma muscle injected with botulinum toxin can improve the lateral cheek wrinkles and marionette lines, lifting the lower face (see, e.g., Tamura, B.M., The Effect of Botulinum Toxin on the Platysma Muscle, Curr. Derm. Rep. 2012, 1:89-95; Kane et al., Expanding the Use of Neurotoxins in Facial Aesthetics: A Consensus Panel’s Assessment and Recommendations, J. Drugs Dermatol. 2010, 9(1):s7-s25).

[0007] However, the use of botulinum toxin in the lower face and neck is more complex than in the upper face and thus presents more complications. This is because the muscles in this area are very compact and interact at different levels and depths to perform different functions such as speaking, eating, drinking, and facial expressions. The injected botulinum toxin may spread or diffuse to the swallowing muscles, pharynx, and neck flexor structures, and cause complications such as dysphagia (difficulty swallowing), dysphonia, and neck weakness (see, e.g., De Boulle et al., Chapter 15: Neck Rejuvenation, in: Botulinum Toxins: Cosmetic and Clinical Applications, Cohen, J.L., Ozog, D.M., and Porto, D.A. (eds.), Wiley-Blackwell, 2017; Chen, D.L. and Cohen, J.L., Botulinum Toxin-A Chemical Denervation for Platysmal Bands: Maximal Dosing Considerations, J. Drugs Dermatol. 2015, 14(9):931; Carruthers et al., Consensus Recommendations on the Use of Botulinum Toxin Type A in Facial Aesthetics, Plast. Reconstr. Surg. 2004, 114(6 Suppl):1S-22S)).

[0008] These adverse side effects can be reduced by using lower doses. However, the effects of lower doses are less effective and may result in the formation of residual bands and the development of compensatory bands due to incomplete paralysis of the platysma muscle. In addition, when using lower doses, the duration of action is shorter and more frequent treatments are required.

[0009] In multiple studies, the relationship between toxin dilution (concentration, volume) and efficacy and safety has been investigated, but the results are controversial. For example, it has been found that dilution increases the incidence of side effects observed in the treatment of glabellar lines, but has no significant effect on efficacy or duration (Carruthers et al., Dilution volume of botulinum toxin type A for the treatment of glabellar rhytides: does it matter?, Dermatol. Surg. 2007, 33: S97 - S104). In another study on dynamic forehead lines (Hsu et al., Effect of volume and concentration on the diffusion of botulinum exotoxin A, Arch. Dermatol. 2004, 140(11): 1351 - 1354), it was found that larger volumes are beneficial as they result in greater diffusion and a larger affected area. The authors concluded that for treating larger confluent areas, such as the forehead, larger volumes can be used to achieve greater diffusion. This also means fewer injections are required, which reduces the pain during injection and increases patient comfort and satisfaction. On the other hand, the authors noted that greater dilution tends to have a shorter duration of action and carries the risk of migration to unintended muscles.

[0010] Regarding the treatment of platysma bands, multiple studies have shown that botulinum toxin treatment is an alternative, effective, and non-invasive method for treating platysma bands (see, for example, Brandt, F.S. and Bellman, B., Cosmetic Use of Botulinum A Exotoxin for the Aging Neck, Dermatol. Surg. 1998, 24:1232-1234; Matarasso et al., Botulinum A Exotoxin for the Management of Platysma Bands, Plast. Reconstr. Surg. 1999, 103(2):645-652; Kane, M., Nonsurgical Treatment of Platysmal Bands with Injection of Botulinum Toxin A, Plast. Reconstr. Surg. 1999, 103:656-665; Brandt, F.S. and Boker, A., Botulinum Toxin for Rejuvenation of the Neck, Clin. Dermatology 2003, 21:515-520; Kane et al., Expanding the Use of Neurotoxins in Facial Aesthetics: A Consensus Panel’s Assessment and Recommendations, J. Drugs Dermatol. 2010, 9(1):s7-s25; Gubanova et al., Efficacy and Safety of IncobotulinumtoxinA for the Treatment of Platysmal Bands of the Aging Neck: An Open-Label, Prospective Pilot Study, J. Drugs Dermatol. 2013, 12(12):1461-1466; Prager et al., IncobotulinumtoxinA for the Treatment of Platysmal Bands: A Single-Arm, Prospective Proof-of-Concept Clinical Study, Dermatol. Surg.2015, 41: S88 - S92; see also the review by Sugrue et al., Botulinum Toxin Treatment for Mild to Moderate Platysma Bands: A Systematic Review of Efficacy, Safety, and Injection Technique, Aesthet. Surg. J. 2019, 39(2): 201 - 206).

[0011] In the published literature, significantly different botulinum toxin doses for treating platysma bands have been reported. OnabotulinumtoxinA with a total dose between 10U and 100U (see Brandt and Bellman, 1998, supra; Kane, 1999, supra; Brandt and Boker, 2003, supra; Tamura, 2012, supra; Chen and Cohen, 2015, supra) and up to 250U (see Matarasso et al., 1999, supra) have been used for successful treatment of platysma bands. In addition, a total dose of 30U to 60U of ncobotulinumtoxin A has been described (see Gubanova et al., 2013; supra; Prager et al., 2015, supra; see also the review by Sugrue et al., supra), and a total dose of 5 - 20U per band of abobotulinumtoxinA (DysportTM) has been described (Kane et al., 2010, supra). In addition, according to the consensus recommendations on the use of botulinum toxin type A in facial aesthetics (see Carruthers et al., 2004, supra), the dose per injection site is also quite variable, resulting in a total dose per band of 6 to 40+U, depending on the number of injection sites (usually 3 to 5) and the dose used.

[0012] In addition, a variety of different diluents are used in the art. For example, Matarasso et al., 1999 (see above) used 5-10 U of onabotulinumtoxin A in 0.1-0.2 mL at each injection site. Prager et al., 2015 (see above) injected 2.5 U of incobotulinumtoxin A (total amount of 60 U) in an injection volume of 0.0625 mL at each injection site. Gubanova et al., 2013 (see above) injected 5 U of incobotulinumtoxin A in 0.1 mL at each injection site, with a total amount of 60 U. In addition, as disclosed in the textbook "Illustrated Guide to Aesthestic Botulinum Toxin Injections" (Kane M. and Sattler, g., 2013, Quintessence Publishing Co Ltd.), the recommended dose for the platysma band is 1-2 U of incobotulinumtoxin A and onabotulinumtoxin A in 0.025 to 0.05 mL at each injection site.

[0013] The aging neck is characterized not only by the platysma band but also by the loss of the mandibular contour (the mandibular part of the platysma). After treatment of the platysma band, overall neck tightening, reshaping of the mandibular contour (Brandt and Bellmann, 1998, see above; Tamura, 2012, see above) or lower face lift (Kane et al., 2010, see above) have been observed. Levy et al. paid particular attention to the mandibular contour and introduced a new technique called the "Nefertiti lift" (see Levy, P.M., The 'Nefertiti lift': A new technique for specific re-contouring of the jawline, J. Cosm. Laser Ther. 2007, 9: 249-252). In addition to treating the platysma band, Levy also injected botulinum toxin type A into the platysma part behind the nasolabial fold, along and below the mandible. Injecting botulinum toxin type A releases the tension on the mandibular contour by reducing the downward pressure of the platysma, so that the elevator muscle predominates, resulting in skin lifting and reshaping of the mandibular contour.

[0014] de Almeida et al. disclosed another injection pattern (de Almeida et al., The Facial Platysma and Its Underappreciated Role in Lower Face Dynamics and Contour, Dermatol. Surg. 2017, 43:1042-1049), which not only addressed the issues of the lateral part of the platysma in the mandible described by Levy et al. (see above), but also treated the anterior fibers of the platysma and the mentalis muscle. This treatment pattern led to remodeling of the mandibular contour and improvement of the lateral buccal lines and the horizontal lines along the mandible under the mandible. Bravo et al. (Bravo et al., J. Clin. Aesthet. Dermatol. 2019, 12(11):32-34) described another technique for improving the mandibular contour and definition.

[0015] Despite these advancements, there is still a need in the art for an improved rejuvenation treatment of the neck, optionally together with the lower face, with botulinum toxin that produces highly satisfactory cosmetic effects without endangering the safety of the patient.

[0016] Object of the present invention

[0017] Accordingly, it is an object of the present invention to provide an efficient and safe botulinum toxin therapy for neck rejuvenation. Summary of the Invention

[0018] The present invention is based on the discovery that administering a high-concentration botulinum toxin preparation to the platysma muscle, more specifically, to the platysma bands and optionally to the platysma at the mandibular contour, can effectively and safely rejuvenate the neck or optionally rejuvenate the neck and the lower face. This mode of administration localizes the toxin to the target platysma muscle to provide the desired cosmetic effect and prevents the toxin from migrating to non-target muscles. In the neck, anatomical structures such as the swallowing muscles, the larynx, and the neck flexors are very close, and a small migration potential is crucial for avoiding diffusion to these important anatomical structures. The "high-concentration" (or "low-volume-high-dose") method of the present invention avoids such complications while providing high efficacy.

[0019] In a first aspect, the present invention relates to the use of botulinum toxin for neck rejuvenation, wherein the botulinum toxin is injected at a concentration greater than 100 U / ml, preferably at a concentration of 150 U / ml or higher, into at least one platysma band of a subject.

[0020] Preferably, the dose injected at each injection point in at least one platysma band is at least 3 U, more preferably 3 U to 5 U, such as 3 U, 4 U or 5 U. The volume at each injection point is preferably 0.030 ml or less, especially 0.010 ml to 0.025 ml. In the present invention, a high-concentration botulinum toxin solution is used. It has been found that administering a high dose of botulinum toxin in a low volume is particularly suitable for thin muscles such as the platysma.

[0021] According to the present invention, botulinum toxin can be additionally injected into the platysma at the mandibular contour on both sides of the face to enhance the rejuvenating effect of the injection into the platysma band. By relaxing the platysma along the mandibular contour, the downward pull of the platysma will be counteracted and the elevator muscles will be dominant, resulting in an additional lifting effect and reshaping of the mandibular contour.

[0022] In a second aspect, the present invention relates to a method for neck rejuvenation, wherein botulinum toxin is injected at a concentration greater than 100 U / ml, preferably at a concentration of 150 U / ml or higher, into at least one platysma band of a subject.

[0023] Preferred embodiments are set forth in the appended dependent claims and in the following detailed description in conjunction with the examples and drawings provided therein. Description of the Drawings

[0024] Figure 1 Diffusion of toluidine dye in porcine muscle after intramuscular injection is described. (A): Injection of 10 μl, 20 μl or 40 μl of toluidine dye at a 45° injection angle; (B) Injection of 10 μl, 20 μl or 40 μl of toluidine dye at a 90° injection angle.

[0025] Figure 2 An exemplary injection pattern according to the present invention is shown for improving the prominence of the platysma band and the sharpness of the mandibular contour. Detailed Description

[0026] The high-concentration method (or high-dose / low-volume concept) used in the present invention allows the skilled person to safely administer botulinum toxin at a higher dose at each injection point, thereby producing excellent efficacy in terms of the duration of effectiveness in the treatment of the platysma.

[0027] A low volume is believed to prevent the spread of toxins to adjacent muscles, thereby preventing complications and unwanted side effects. This is particularly important in the case of the platysma muscle, as the platysma muscle is rather thin and anatomically close to important structures. Therefore, special attention is required to prevent complications associated with treating the platysma bands with botulinum toxin. The low volume used according to the present invention reduces the risk of migration / spread to unintended muscles, thereby eliminating the main concern in using botulinum toxin in cosmetic indications, especially in the treatment of the platysma muscle.

[0028] The low volume method allows for the use of higher toxin doses per injection site as well as a higher total dose, which improves the efficacy of the treatment. In addition, the higher dose will prolong the duration of action, resulting in longer treatment intervals and thus increased patient satisfaction. Further, according to the present invention, both the platysma muscle along the mandibular border and the platysma bands can be treated simultaneously in one treatment to achieve complete relaxation of the platysma muscle. It is expected that additional treatment of the platysma muscle in the mandibular contour area reduces the formation of residual bands and the development of compensatory bands.

[0029] Overall, the present invention allows for the treatment of the platysma muscle as an anatomical and cosmetic entity and achieves a balanced and harmonious appearance of the neck, increasing patient satisfaction without increasing side effects. In addition, it enables the use of longer treatment intervals, resulting in a lower immunogenic risk of neutralizing antibody formation (known neutralizing antibody formation can lead to secondary treatment failure), thereby increasing patient convenience and satisfaction. Thus, low volume and high dose botulinum toxin treatment (i.e., using high concentrations of botulinum toxin) provides high efficacy, excellent aesthetic results, and long-lasting effects while preventing or minimizing unwanted side effects.

[0030] In a first aspect, the present invention relates to the use of botulinum toxin for neck rejuvenation, wherein the botulinum toxin is injected at a concentration greater than 100 U / ml, preferably at a concentration of 150 U / ml or higher, into at least one platysma band of a subject.

[0031] As used herein, the term "rejuvenation" broadly refers to restoring a youthful appearance. In addition, when used in connection with the uses and methods of the present invention, the term "rejuvenation" refers to a purely aesthetic application. According to the present invention, botulinum toxin is used only for cosmetic purposes.

[0032] As used herein, the term "lower face" refers to the area from subnasale to menton, including the mandibular contour (including jowls and marionette lines). As used herein, the term "neck" refers to the area extending from the lower part of the mandible (lower jawbone) down to the clavicles and sternum.

[0033] As used herein, the term "platysma" has the same meaning as the term "platysma muscle", and thus these terms are used interchangeably herein. The platysma is a thin layer of muscle that wraps around the entire neck. It originates from the fascia covering the deltoid and upper pectoral muscles, and a portion of the platysma extends across the mandibular margin and inserts into the lower face just above the mandible. Thus, since only the platysma is the target in the present invention, an injection into the mandibular contour should be understood as an injection into the platysma at the mandibular contour. Since this portion of the platysma inserts into the lower face, these injections are intended to rejuvenate the lower face.

[0034] As used herein, the term "platysmal band" has the meaning known in the art and refers to a bundle or band in the neck. As used herein, the term "platysma" or the term "platysma" encompasses the platysmal band. Platysmal bands are a feature of the aging neck. During the aging process, the platysma begins to separate and bunch up into bundles or bands, typically alongside the midline on each side of the neck and on the sides of the neck. Thus, platysmal bands can be and typically are medial bands and lateral bands, with two medial bands and two lateral bands. In the context of the present invention, the term "injection into the platysmal band" refers to an intramuscular injection into the platysma. Injecting botulinum toxin into the platysmal band can relax and flatten the platysmal band, making the band less visible. It should be noted that as used herein, the term "platysmal band" refers to a vertical band, i.e., a band that runs vertically through the front of the neck. Thus, a "platysmal band" is different from a necklace wrinkle, i.e., a horizontal band or wrinkle that extends horizontally around the front of the neck.

[0035] As used herein, the term "subject" is not particularly limited and refers to a person in need of cosmetic treatment of the neck or the neck and lower face. In the context of the present invention, the term "subject" can be used interchangeably with the term patient.

[0036] In the context of the present invention, a numerical value without decimal places should be understood to include all numerical values having one or more decimal places, applying the general rounding rules and giving the numerical value without decimal places. For example, the numerical value 3 includes 2.5 or 2.50 (rounded up to 3) and 3.4 or 3.49 (rounded down to 3), and all numerical values in between. The same applies to the endpoints of a range, i.e., a range from 3 to 5 can mean, for example, 2.5 - 3.4 to 4.5 - 5.4. It is also contemplated that one or both (preferably both) endpoints of each numerical value or range disclosed in this application, if they have no decimal places, can be numerical values with zero decimal places, e.g., 4 can be 4.0.

[0037] As used herein, the term "botulinum toxin" is not particularly limited and includes botulinum toxins of any serotype (BoNT / A-H). For example, the botulinum toxin may be serotype A or B (BoNT / A, BoNT / B). Preferably, the botulinum toxin is serotype A, more preferably serotype A1 (BoNT / A1), and most preferably BoNT / A1 produced by Clostridium botulinum Hall strain. In addition, as used herein, the term "botulinum toxin" ("BT") and the synonymous term "botulinum neurotoxin" ("BoNT") refer to pure botulinum neurotoxin and / or any complex thereof, i.e., any complex of pure botulinum neurotoxin and complexing protein (referred to as "toxin complex"). Preferably, the botulinum toxin is a pure botulinum neurotoxin of serotype A.

[0038] As used herein, the term "pure botulinum neurotoxin" refers to botulinum neurotoxin without complexing protein (sometimes also referred to as "neurotoxic component"), or more precisely, botulinum neurotoxin without neurotoxin-associated complexing protein (NAP). Pure botulinum neurotoxin is the (active) neurotoxic polypeptide that ultimately inhibits acetylcholine release. It is a double-chain protein comprising a light chain (LC; approximately 50 kDa) and a heavy chain (HC; approximately 100 kDa) bound together by a disulfide bond. Thus, the active neurotoxic polypeptide may also be referred to herein as "150 kDa neurotoxin", "botulinum neurotoxin (150 kD)" or "neurotoxic component". Preferably, the botulinum toxin is the (pure) botulinum neurotoxin contained in or is

[0039] The term "toxin complex" as used herein refers to a high molecular weight complex of the neurotoxic component and a group of complexing proteins (NAP), including type A botulinum toxin complexes of 900 kDa, 500 kDa and 300 kDa. The complexing proteins are non-toxic non-hemagglutinins (NTNHA), which are different hemagglutinins (HA) in A-D serotype strains. For example, the 900 kDa complex is contained in onabotulinumtoxin A ( Allergan, Inc., Irvine, CA, USA), and abobotulinumtoxin A ( Ipsen, Paris, France), (lpsen / Galderma) and (Medytox) also contain the toxin complex as the active agent. Preferably, in addition to the pure botulinum neurotoxin contained in or is the botulinum toxin is the one contained in or the toxin complex in, or or In another preferred embodiment, the botulinum toxin is not contained in the toxin complex or is not

[0040] The botulinum toxin may be a natural neurotoxin obtainable from the bacterium Clostridium botulinum or any other botulinum toxin, such as a botulinum toxin obtainable from alternative sources, including recombinant technology and genetic or chemical modification. Chimeric or genetically modified botulinum toxins, i.e., botulinum toxins containing mutations including substitutions, deletions, and insertions, are also included within the terms "botulinum toxin", "neurotoxic component", etc. Preferably, the mutations do not impair any biological activity of the botulinum toxin. However, the use of mutations to modulate the biological activity of the botulinum toxin is also contemplated. Also included are botulinum toxins containing chemically modified amino acids, such as one or more glycosylated, acetylated, or otherwise modified amino acids, which may be beneficial for toxin uptake or stability. Particularly preferred is the lipidation of the neurotoxic component.

[0041] In the present invention, the dose is expressed in biological units because the botulinum toxin used may contain, for example, different percentages of inactive toxin, which contributes to the total protein load but not to the efficacy. In the context of the present invention, the mouse bioassay (MBA) is used to determine the biological potency of the botulinum toxin. The MBA determines the median lethal dose (LD50) of the toxin / neurotoxin after intraperitoneal injection in mice, i.e., the dose of the toxin / neurotoxin that is capable of killing 50% of a group of mice. On this basis, as used herein, 1 unit (U) of the toxin / neurotoxin is defined as one mouse LD50 (1.0 LD50 = 1.0 U). The LD50 mouse bioassay is the gold standard among the various biological, chemical, or immunological assays for botulinum toxin and is known to those skilled in the art (see, e.g., Pearce, L.B.; Borodic, G.E.; First, E.R.; MacCallum, R.D. Measurement of botulinum toxin activity: Evaluation of the lethality assay. Toxicol. Appl. Pharmacol. 1994, 128, 69 - 77).

[0042] Another useful method for determining the biological activity (biological potency) of botulinum neurotoxin is a cell-based potency assay, as disclosed, for example, in WO2009 / 114748, WO 2013 / 049508 or WO 2014 / 207109. The activity results obtained with this cell-based assay correspond to the activity values obtained with a mouse intraperitoneal (i.p.) LD50 assay, since these values are calibrated using an LD50 reference standard.

[0043] Due to the differences in the LD50 tests used by manufacturers of commercial botulinum toxin preparations, the unit potencies labeled by manufacturers for their commercial botulinum toxin preparations are proprietary and not easily comparable. Thus, within the framework of the present invention, the conversion rates provided below are used to establish the relative potencies of incobotulinumtoxinA ("INCO"; botulinum toxin serotype A, without complexing protein; Merz Pharmaceuticals GmbH), onabotulinumtoxinA ("ONA"; botulinum toxin complex of serotype A; Allergan Inc.), abobotulinumtoxinA ("ABO"; botulinum toxin complex of serotype A; Medicis Pharmaceutical Corp., Galderma Lab.), rimabotulinumtoxinB ("RIM"; botulinum toxin serotype B; Solstice Neurosciences Inc.) and ("TBD"; botulinum toxin serotype A; Mentor Worldwide LLC). In the present context, the conversion rate of ONA and INCO is 1:1. The conversion rate of ONA / INCO:ABO is 1:2.5. The conversion rate of ONA / INCO:RIM is 1:50, and the conversion rate of ONA / INCO:TBD is 1:1.5. Further preferably, within the context of the present invention, 1U of INCO and 1U of onabotulinumtoxinA ("ONA"; ) should be considered as corresponding to one mouse LD50 (1.0 LD50) or 1U measured by the above method (i.e., using MBA). Even more preferably in the present invention, if the botulinum toxin is abobotulinumtoxinA, 2.5U of abobotulinumtoxinA is equal to 1U, and 1U is preferably one mouse LD50 (1.0 LD50) measured using the above MBA.

[0044] In a preferred embodiment, a first aspect of the present invention relates to the use of botulinum toxin for neck rejuvenation, wherein the botulinum toxin is injected at a concentration greater than 100 U / ml into at least one platysma band of a subject, provided that if the botulinum toxin is abobotulinumtoxinA, 2.5 U of abobotulinumtoxinA is equal to 1 U. In another preferred embodiment, a first aspect of the present invention relates to the use of botulinum toxin for neck rejuvenation, wherein the botulinum toxin is injected at a concentration greater than 100 U / ml into at least one platysma band of a subject, wherein 1 U corresponds to 1 LD50, and LD50 is the median lethal dose measured using the mouse LD50 bioassay, provided that if the botulinum toxin is abobotulinumtoxinA, 2.5 U of abobotulinumtoxinA is equal to 1 U. In another preferred embodiment, a first aspect of the present invention relates to the use of botulinum toxin for neck rejuvenation, wherein the botulinum toxin is injected at a concentration greater than 100 U / ml into at least one platysma band of a subject, wherein 1 U corresponds to 1 LD50, and LD50 is the median lethal dose measured using the mouse LD50 bioassay, provided that if the botulinum toxin is incobotulinumtoxinA or onabotulinumtoxinA, 1 U of incobotulinumtoxinA and 1 U of onabotulinumtoxinA are both equal to 1 U, and if the botulinum toxin is abobotulinumtoxinA, 2.5 U of abobotulinumtoxinA is equal to 1 U.

[0045] As used herein, the term "comprising", and the terms "including", "containing", and any variants thereof, such as "comprises", "includes", and "contains", are intended to mean non-exclusive inclusion, such that a process, method, product obtained by the process, composition, or formulation that comprises, includes, or contains one element or a series of elements not only includes those elements, but may also include other elements not expressly listed for such process, method, product obtained by the process, composition, or formulation. Furthermore, within the framework of the present invention, it is intended that each of the terms "comprising", "including", "containing", and any variants thereof may be replaced by the term "consisting of" or any of its variants, which shall be understood to refer to the exclusive inclusion of the indicated elements.

[0046] The terms "a", "an", and "the" and similar references used in the context of the present invention shall be construed to cover both the singular and the plural, and thus may also refer to "at least one" or "more than one", unless otherwise specified herein or clearly contradicted by the context.

[0047] According to a first aspect of the present invention, botulinum toxin is injected at a concentration greater than 100 U / ml, preferably 125 U / ml or higher, 150 U / ml or higher, 175 U / ml or higher, 200 U / ml or higher, 225 U / ml or higher, or 250 U / ml or higher into at least one platysma band of a subject. The upper limit is preferably 500 U / ml. Preferably, botulinum toxin is injected at a concentration greater than 100 U / ml to 500 U / ml or 125 U / ml to 400 U / ml, more preferably at a concentration of 125 U / ml to 275 U / ml or 150 U / ml to 350 U / ml or 150 U / ml to 300 U / ml or 150 U / ml to 250 U / ml or 175 U / ml to 275 U / ml or 200 U / ml to 250 U / ml. More preferably, botulinum toxin is injected at a concentration of 130 U / ml to 170 U / ml, or 180 U / ml to 220 U / ml, or 230 U / ml to 270 U / ml, and most preferably at a concentration of 150 U / ml, 200 U / ml or 250 U / ml.

[0048] In addition, in addition to injecting the botulinum toxin concentrations disclosed herein, preferably botulinum toxin is injected at a dose of at least 2 U (e.g., 2 U to 6 U, such as 3 U, 4 U or 5 U) per injection site and a volume of 0.03 ml or less. Preferably, botulinum toxin is injected at a dose of at least 3 U per injection site, particularly at a dose of 3 U to 6 U (e.g., a dose of 3 U, 4 U, 5 U or 6 U) and a volume of less than 0.030 ml. More preferably, botulinum toxin is injected at a dose of at least 3 U per injection site, particularly at a dose of 3 U to 6 U (e.g., a dose of 3 U, 4 U, 5 U or 6 U) and a volume of 0.010 ml to 0.025 ml, and more particularly at a dose of at least 3 U per injection site, particularly at a dose of 3 U to 6 U (e.g., a dose of 3 U, 4 U, 5 U or 6 U) and a volume of 0.015 ml to 0.025 ml.

[0049] Most preferably, botulinum toxin is injected at a dose of 3 U to 5 U per injection site and a volume of less than 0.030 ml, particularly at a dose of 3 U to 5 U per injection site and a volume of 0.010 to less than 0.030 ml, more particularly at a dose of 3 U to 5 U per injection site and a volume of 0.010 ml to 0.025 ml, and most particularly at a dose of 3 U to 5 U per injection site and a volume of 0.015 ml to 0.025 ml.

[0050] According to the present invention, botulinum toxin is injected into one or more platysma bands, preferably into 2 or 4 platysma bands, more preferably into the two medial bands on the left and right or the two lateral bands on the left and right, and most preferably into 4 platysma bands, i.e., each of the left medial band, the right medial band, the left lateral band, and the right lateral band. In other cases, such as asymmetric bands, botulinum toxin can also be injected into 3 platysma bands.

[0051] Generally, botulinum toxin is injected into at least three injection points in each platysma band, preferably into 4, 5, or 6 injection points in each platysma band. The total number of injection points in the platysma bands (i.e., all platysma bands) is generally 6 to 24, preferably 8 to 22, more preferably 14 to 20, and most preferably 16 to 20.

[0052] The total volume of botulinum toxin injected into the at least one platysma band (i.e., all treated platysma bands) is generally 0.15 ml to 0.60 ml, preferably 0.20 ml to 0.50 ml, more preferably 0.30 ml to 0.45 ml or 0.35 ml to 0.40 ml, and most preferably 0.40 ml.

[0053] The total dose of botulinum toxin injected into the at least one platysma band (i.e., all treated platysma bands) is generally 25 U to 200 U, preferably 30 U to 150 U or 40 U to 120 U, more preferably 50 U to 120 U or 60 U to 110 U, and most preferably 60 U to 100 U or 80 U to 100 U.

[0054] As used herein, the terms "total dose" and "total volume" respectively refer to the total dose and total volume of botulinum toxin that are generally administered during the treatment period (or "injection period") and are administered by multiple injections at different injection points in a single area. The treatment period generally lasts less than one hour, for example, about 30 minutes. The single area is the platysma band to be treated, and optionally, the platysma at the mandibular contour.

[0055] The distance between the injection points along the platysma band is about 1 cm to about 3 cm, preferably about 1.5 cm to about 2.5 cm, and more preferably about 2 cm. This interval refers to the distance between adjacent injection points on the platysma band.

[0056] According to the present invention, botulinum toxin can also be injected and preferably is injected into the platysma at the mandibular contour of the subject. In other words, the use according to the present invention includes the injection of the platysma band as described above, and optionally, the injection of botulinum toxin into the platysma at the mandibular contour. The injection at the mandibular contour releases the tension on the mandibular contour by reducing the downward pressure of the platysma, so that the elevator muscles are dominant, resulting in skin lifting and reshaping of the mandibular contour.

[0057] As described above, the platysma muscle passes through the mandible, with some of its fibers inserting into the bone below the oblique line, and other fibers extending approximately 4 cm above the mandibular margin, entering the cheek, and inserting into the skin and subcutaneous tissue. Thus, as used herein, the term "injection in the mandibular contour" or "injection at the mandibular contour" or similar terms are intended to mean an injection into the platysma muscle. As a result, the platysma muscle will be antagonized and the elevator muscles will be dominant, resulting in an additional lifting effect and reshaping of the mandibular contour. Further, unless otherwise specified, the term "injection in the mandibular contour" or "injection at the mandibular contour" or similar terms preferably refer to an injection into each or both mandibular contours of the subject's face.

[0058] Preferably, botulinum toxin is injected into the platysma muscle at the mandibular contour at a concentration of at least 50 U / ml per injection site, preferably at least 100 U / ml or at least 150 U / ml or at least 200 U / ml. Most preferably, botulinum toxin is injected into the platysma muscle at the mandibular contour at a concentration of 100 U / ml to 250 U / ml, such as at a concentration of 150 U / ml, 200 U / ml, or 250 U / ml.

[0059] In addition, in addition to or as an alternative to the botulinum toxin concentration related to the mandibular contour described above, within the context of the present invention, botulinum toxin is preferably injected into the platysma muscle at the mandibular contour at a dose of at least 1 U per injection site, particularly 1 U to 4 U per injection site, more particularly 1 U to 3 U or 1.5 U to 2.5 U (such as 1.5 U, 2 U, or 2.5 U), and most particularly 2 U per injection site.

[0060] Specifically, botulinum toxin is injected into the platysma muscle at the mandibular contour at a dose of at least 1 U per injection site, preferably 1 U to 4 U per injection site, more preferably 1 U to 3 U or 1.5 U to 2.5 U per injection site, such as 1.5 U, 2 U or 2.5 U, most preferably 2 U per injection site, and at a concentration of at least 100 U / ml per injection site. More preferably, botulinum toxin is injected into the platysma muscle at the mandibular contour at a dose of at least 1 U per injection site, preferably 1 U to 4 U per injection site, more preferably 1 U to 3 U or 1.5 U to 2.5 U per injection site, such as 1.5 U, 2 U or 2.5 U, most preferably 2 U per injection site, and at a concentration of at least 150 U / ml per injection site. Most preferably, botulinum toxin is injected into the platysma muscle at the mandibular contour at a dose of at least 1 U, preferably 1 U to 4 U, more preferably 1 U to 3 U or 1.5 U to 2.5 U, such as 1.5 U, 2 U or 2.5 U, most preferably 2 U, and at a concentration of at least 200 U / ml (such as 200 U / ml or 250 U / ml) per injection site. The upper limit of all the above open ranges is preferably 500 U / ml, most preferably 400 U / ml. A person skilled in the art can easily select a suitable concentration according to the dose to be injected and considering that for practical reasons and to avoid inaccurate dosing, the injection volume should not be too low.

[0061] Preferably, botulinum toxin is injected into the platysma muscle at the mandibular contour at a volume of 0.005 ml to 0.03 ml per injection site, more preferably at a volume of 0.01 ml to 0.025 ml per injection site, such as 0.02 ml or 0.01 ml, and most preferably at a volume of 0.01 ml per injection site.

[0062] Furthermore, within the context of the present invention, preferably botulinum toxin is injected into the platysma muscle at the mandibular contour at a dose of at least 1 U per injection site, in particular 1 U to 4 U per injection site, more particularly 1 U to 3 U or 1.5 U to 2.5 U per injection site, such as 1.5 U, 2 U or 2.5 U, at a volume of 0.01 ml to 0.02 ml.

[0063] Furthermore, within the context of the present invention, preferably botulinum toxin is injected into the platysma muscle at the mandibular contour at a dose of at least 1 U per injection site, in particular 1 U to 4 U per injection site, more particularly 1 U to 3 U or 1.5 U to 2.5 U per injection site, such as 1.5 U, 2 U or 2.5 U, at a volume of 0.01 ml.

[0064] Furthermore, within the context of the present invention, botulinum toxin is preferably injected in the platysma muscle at the mandibular contour at a dose of at least 1 U per injection site, in particular 1 U to 4 U per injection site, more particularly 1 U to 3 U or 1.5 U to 2.5 U per injection site, such as 1.5 U, 2 U or 2.5 U, most preferably 2 U, in a volume of 0.02 ml.

[0065] The total volume of botulinum toxin injected in the platysma muscle at each mandibular contour can be from 0.02 ml to 0.20 ml, in particular from 0.02 ml to 0.16 ml or from 0.04 ml to 0.16 ml, preferably from 0.06 ml to 0.12 ml, more preferably from 0.08 ml to 0.12 ml, most preferably from 0.08 ml to 0.10 ml. The total dose of botulinum toxin injected in the platysma muscle at each mandibular contour can be from 4 U to 16 U, preferably from 5 U to 12 U, more preferably from 6 U to 10 U (such as 6 U, 8 U or 10 U), most preferably 8 U.

[0066] The number of injection sites in the platysma muscle at each mandibular contour is at least 2, preferably 2 to 6, more preferably 3 to 5, most preferably 4. Furthermore, the distance between the injection sites along the mandibular contour is about 0.5 cm to about 2 cm, preferably about 1.0 cm to about 1.5 cm, more preferably about 1 cm. Preferably, botulinum toxin is administered to 2 to 4 injection sites on each side along each mandible and under each mandible, rather than in front of the line connecting the nasolabial fold down to the mandible.

[0067] For the use according to the first aspect of the present invention, botulinum toxin can be administered in consecutive treatment cycles. It should be understood that a treatment cycle is the time interval between two administrations of botulinum neurotoxin, i.e., a treatment cycle consists of an administration of botulinum neurotoxin and the subsequent period until the next administration of botulinum neurotoxin injection. The administration of the botulinum toxin occurs during the treatment period (or "injection period"). The time interval between two consecutive administrations of botulinum neurotoxin can vary between about 5 and 10 months, in particular between about 6 and 9 months or between about 7 and 8 months. The longer duration of the effect achieved by the present invention makes it possible to extend the treatment interval.

[0068] According to the present invention, botulinum toxin (i.e., the neurotoxic component or toxin complex) can be serotype A, B, C1, D, E, F, G or H. Since botulinum toxin type A has a more persistent effect compared to other serotypes, botulinum toxin type A is preferably used herein. Furthermore, within the framework of the present invention, the term "botulinum toxin" can refer to the natural neurotoxin obtainable from the bacterium Clostridium botulinum or botulinum toxin obtainable from alternative sources, including botulinum toxin obtained from recombinant technology or from genetic or chemical modification.

[0069] Generally, the botulinum toxin used in the present invention exists in the form of a liquid composition. As is known in the art, liquid compositions can be formulated by various techniques according to the desired application. It can be provided as a ready-to-use liquid preparation or in the form of a lyophilized powder that is usually reconstituted in physiological saline before use. Preferably, the botulinum toxin used in the present invention is in the form of an aqueous solution, more preferably a saline solution or a physiological saline solution, and most preferably a phosphate-buffered saline solution. The aqueous solution can additionally contain one or more pharmaceutically acceptable substances. Suitable pharmaceutically acceptable substances include those well known in the art, see for example Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania.

[0070] Specifically, the aqueous botulinum toxin solution or composition can contain other carriers or non-toxic, non-therapeutic, non-immunogenic stabilizers, etc. Thus, the aqueous botulinum toxin composition can contain glycerol, a protein stabilizer (HSA) or a non-protein stabilizer such as polyvinylpyrrolidone (PVP), hyaluronic acid or free amino acids such as methionine or histidine. The aqueous botulinum toxin composition can be free of amino acids. In one embodiment, it can be free of stabilizing peptides (e.g., consisting of 5 to 50 amino acids, 10 to 40 amino acids or 15 to 30 amino acids). Non-protein stabilizers suitable for use herein are disclosed in WO 2005 / 007185 or WO 2006 / 020208. The botulinum toxin composition can also contain non-ionic or ionic surfactants such as polysorbates or poloxamers. Suitable formulations for HSA-stabilized formulations containing the botulinum toxin according to the present invention are disclosed for example in US 8,398,998 B2.

[0071] Preferably, the botulinum toxin used in the present invention exists in the form of an aqueous solution containing sodium chloride (NaCl), more preferably in the form of a physiological saline solution (i.e., a solution containing physiological concentration of sodium chloride, e.g., about 9 g / l NaCl), wherein the aqueous botulinum toxin solution contains (i) no other excipients (except NaCl), (ii) human serum albumin (HSA) and sugar, especially monosaccharide or disaccharide, (iii) human serum albumin (HSA) and lactose, (iv) human serum albumin (HSA) and sucrose, (v) monosaccharide and / or disaccharide (e.g., lactose and / or sucrose), (vi) no buffer, (vii) no single amino acid, (viii) no human serum albumin (HSA), sodium chloride and lactose or no HSA, sodium chloride and sucrose, or (ix) no HSA and sodium chloride, or any combination of (i) to (ix).

[0072] In a second aspect, the present invention relates to a method for neck rejuvenation, the method comprising injecting botulinum toxin at a concentration greater than 100 U / ml into at least one platysma band of a subject.

[0073] This treatment method is purely for aesthetic purposes, i.e., for the rejuvenation of the neck or the neck and the lower face, and thus is a cosmetic method. Botulinum toxin is usually intramuscularly injected using a suitable injection device, such as a syringe equipped with a 30G, 32G or 33G needle. The syringe is not particularly limited, and any conventional syringe can be used. A particularly suitable syringe for use according to the present invention is, for example, the 3Dose TM High-precision syringe (vlowmedical).

[0074] The positions, numbers and spacings of the injection points applicable within the framework of the present invention are as described above. Figure 2 An exemplary injection pattern is shown in which, according to this pattern, five injections are made at intervals of 1-2 cm in the left and right medial bands and the left and right lateral bands, so that a total of 20 injections into the platysma bands are made. In addition, 4 injections are made into the platysma at the left and right mandibular contours, i.e., a total of 8 injections. According to this exemplary injection pattern, an improvement in the prominence of the platysma bands and the sharpness of the mandibular contours is obtained.

[0075] The dosing regimen used in combination with the method according to the second aspect of the present invention is as disclosed above with respect to the use according to the first aspect of the present invention. More specifically, the dosing regimen particularly preferred for the present invention (i.e., for the use according to the first aspect and the method according to the second aspect) is as follows:

[0076] Platysma band Mandibular contour (optional)

[0077] BTX concentration: 125 - 300 U / ml 100 - 300 U / ml

[0078] Number of injection points: 16 - 24 (in total) 6 - 10 (in total)

[0079] Dose per injection point: 3 - 5 U 1 - 3 U

[0080] In a preferred embodiment, botulinum toxin is injected at 4 to 6 injection sites per platysma band at a concentration of 150 U / ml to 250 U / ml, wherein the dose per injection site is in the range of 3 U to 5 U, and / or wherein the injection volume per injection site is in the range of 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 3 to 5 injection sites per mandibular contour at a concentration of 150 U / ml to 250 U / ml, wherein the dose per injection site is in the range of 1.5 U to 2.5 U, and / or wherein the injection volume per injection site is in the range of 0.005 ml to 0.015 ml.

[0081] Preferably, botulinum toxin is injected at 4 to 6 injection sites per platysma band at a concentration of 150 U / ml to 250 U / ml, wherein the dose per injection site is in the range of 3 U to 5 U, and wherein the injection volume per injection site is in the range of 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 3 to 5 injection sites per mandibular contour at a concentration of 150 U / ml to 250 U / ml, wherein the dose per injection site is in the range of 1.5 U to 2.5 U, and wherein the injection volume per injection site is in the range of 0.005 ml to 0.015 ml.

[0082] Preferably, botulinum toxin is injected at 4 to 6 injection sites per platysma band at a concentration of 150 U / ml to 250 U / ml, wherein the injection volume per injection site is 0.02 ml, and optionally botulinum toxin is injected at 3 to 5 injection sites per mandibular contour at a concentration of 150 U / ml to 250 U / ml, wherein the injection volume per injection site is 0.01 ml.

[0083] Preferably, botulinum toxin is injected at 20 injection sites on 4 platysma bands at a concentration of 150 U / ml to 250 U / ml, wherein the dose per injection site is in the range of 3 U to 5 U, and / or wherein the injection volume per injection site is in the range of 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 4 injection sites on the platysma at each mandibular contour at a concentration of 100 U / ml to 250 U / ml or 150 U / ml to 250 U / ml, wherein the dose per injection site is in the range of 1.5 U to 2.5 U, and / or wherein the injection volume per injection site is in the range of 0.005 ml to 0.015 ml.

[0084] Preferably, botulinum toxin is injected at 20 injection sites in 4 platysma bands at a concentration of 150 U / ml to 250 U / ml, wherein the dose at each injection site ranges from 3 U to 5 U, and wherein the injection volume at each injection site ranges from 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 4 injection sites in the platysma at each mandibular contour at a concentration of 150 U / ml to 250 U / ml, wherein the dose at each injection site ranges from 1.5 U to 2.5 U, and wherein the injection volume at each injection site ranges from 0.005 ml to 0.015 ml.

[0085] More preferably, botulinum toxin is injected at 20 injection sites in 4 platysma bands at a concentration of 150 U / ml to 250 U / ml, wherein the injection volume at each injection site is 0.02 ml, and optionally botulinum toxin is injected at 4 injection sites in the platysma at each mandibular contour at a concentration of 150 U / ml to 250 U / ml, wherein the injection volume at each injection site is 0.01 ml.

[0086] In a particularly preferred embodiment, botulinum toxin is injected at 4 to 6 injection sites in each platysma band at a concentration of 130 U / ml to 170 U / ml, wherein the dose at each injection site ranges from 2.5 U to 3.5 U, and / or wherein the injection volume at each injection site ranges from 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 3 to 5 injection sites at each mandibular contour at a concentration of 130 U / ml to 170 U / ml, wherein the dose at each injection site is from 1 U to 2 U, and / or wherein the injection volume at each injection site ranges from 0.005 ml to 0.015 ml.

[0087] Preferably, botulinum toxin is injected at 4 to 6 injection sites in each platysma band at a concentration of 150 U / ml, wherein the injection volume at each injection site is 0.02 ml, and optionally botulinum toxin is injected at 3 to 5 injection sites at each mandibular contour at a concentration of 150 U / ml, wherein the injection volume at each injection site is 0.01 ml.

[0088] Most preferably, botulinum toxin is injected at 20 injection sites in 4 platysma bands at a concentration of 150 U / ml, wherein the injection volume at each injection site is 0.02 ml, and optionally botulinum toxin is injected at 4 injection sites in the platysma at each mandibular contour at a concentration of 150 U / ml, wherein the injection volume at each injection site is 0.01 ml.

[0089] In another particularly preferred embodiment, botulinum toxin is injected at 180 U / ml to 220 U / ml at 4 to 6 injection sites per platysma band, wherein the dose at each injection site is in the range of 3.5 U to 4.5 U, and / or wherein the injection volume at each injection site is 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 180 U / ml to 220 U / ml at 3 to 5 injection sites per mandibular contour, wherein the dose at each injection site is 1.5 U to 2.5 U, and / or the injection volume at each injection site is in the range of 0.005 ml to 0.015 ml.

[0090] Preferably, botulinum toxin is injected at 200 U / ml at 4 to 6 injection sites per platysma band, wherein the injection volume at each injection site is 0.02 ml, and optionally botulinum toxin is injected at 200 U / ml at 3 to 5 injection sites per mandibular contour, wherein the injection volume at each injection site is 0.01 ml.

[0091] Most preferably, botulinum toxin is injected at 200 U / ml at 20 injection sites in 4 platysma bands, wherein the injection volume at each injection site is 0.02 ml, and optionally botulinum toxin is injected at 200 U / ml at 4 injection sites in the platysma at each mandibular contour, wherein the injection volume at each injection site is 0.01 ml.

[0092] In another particularly preferred embodiment, botulinum toxin is injected at 230 U / ml to 270 U / ml at 4 to 6 injection sites per platysma band, wherein the dose at each injection site is in the range of 4.5 U to 5.5 U, and / or wherein the injection volume at each injection site is 0.015 ml to 0.025 ml, and optionally botulinum toxin is injected at 230 U / ml to 270 U / ml at 3 to 5 injection sites per mandibular contour, wherein the dose at each injection site is 2 U to 3 U, and / or the injection volume at each injection site is in the range of 0.005 ml to 0.015 ml.

[0093] Preferably, botulinum toxin is injected at 250 U / ml at 4 to 6 injection sites per platysma band, wherein the injection volume at each injection site is 0.02 ml, and optionally botulinum toxin is injected at 250 U / ml at 3 to 5 injection sites per mandibular contour, wherein the injection volume at each injection site is 0.01 ml.

[0094] Most preferably, botulinum toxin is injected at 250 U / ml at 20 injection sites in 4 platysma bands, with an injection volume of 0.02 ml per injection site, and optionally botulinum toxin is injected at 250 U / ml at 4 injection sites in the platysma at each mandibular contour, with an injection volume of 0.01 ml per injection site.

[0095] The extended duration of effect allows for a reduced frequency of administration of botulinum toxin. For example, botulinum neurotoxin can be administered repeatedly, where the time interval between the first and second administrations, or between the second or further administrations and another administration after the second or further administration (i.e., between successive administrations of botulinum toxin) is about 5 to 10 months, particularly about 6 to 9 months or about 7 to 8 months.

[0096] The method according to the second aspect of the invention is closely related to the use according to the first aspect of the invention. Accordingly, all definitions, explanations and advantages given herein with respect to the use according to the first aspect equally apply to the method according to the second aspect.

[0097] Embodiment

[0098] Example 1

[0099] Intramuscular injection of dye into porcine muscle

[0100] A staining dye solution of 0.025% toluidine blue and 0.9% NaCl was injected into porcine lumbar muscle samples (porcine psoas major) using a vlow medical syringe equipped with a 30G x 1 / 2″ subcutaneous injection needle. Three different injection volumes, namely 10 μl, 20 μl and 40 μl, were injected in triplicate at 45° and 90° angles respectively and allowed to diffuse. Then, muscle blocks including the transverse dye diffusion area were cut from the injected porcine psoas major and cross-sections thereof were prepared.

[0101] Figure 1 Photographs of the muscle sections and a scale bar are shown. The muscle sections injected with 10 μl, 20 μl or 40 μl at 45° or 90° angles are presented in (A) and (B) respectively, showing the transverse dye diffusion in a top view.

[0102] It can be seen from Figure 1 that the diffusion depends on the injection volume and occurs along the muscle fibers (even for perpendicular injections). The transverse diffusion of the 10 μl and 40 μl toluidine dye solutions at a 90° injection angle was 10.4 ± 2.2 mm and 23.3 ± 5.4 mm respectively.

[0103] Although these results were obtained using porcine muscle in an ex vivo preclinical model, these results suggest that "low volume" injection of high concentration botulinum toxin at injection sites with a wide interval (e.g., 2 cm distance) in the platysma muscle will provide sufficient distribution to achieve the desired therapeutic effect.

[0104] Example 2

[0105] The injection protocol and dosing regimen of the present invention

[0106] According to the present invention, botulinum toxin can be injected into the platysma muscle using the injection pattern as Figure 2 shown, for improving the prominence of the platysma bands and the sharpness of the mandibular contour. It can be seen that the exemplary injection protocol includes five injections (20 injections in total) in the left and right medial bands and the left and right lateral bands at an interval of 1 - 2 cm and four injections (8 injections in total) in the platysma muscle of the left and right mandibular contours.

[0107] The injected botulinum toxin is typically a 0.9% saline solution. Any suitable injection device, such as a syringe equipped with a 32G needle, can be used to administer the botulinum toxin solution by intramuscular injection into the platysma bands.

[0108] The botulinum toxin solution is administered using the "high concentration method" (high dose / low volume method) according to the present invention. An exemplary dosing regimen using the high concentration method of the present invention is shown in Table 1.

[0109] Table 1. Injection and dosing regimen

[0110]

[0111]

Claims

1. Use of botulinum toxin for neck rejuvenation, wherein the botulinum toxin is injected at a concentration greater than 100 U / ml into at least one platysma band of a subject.

2. The use according to claim 1, wherein the botulinum toxin is injected at a concentration of 150 U / ml or higher.

3. The use according to claim 1 or 2, wherein the botulinum toxin is injected at a dose of at least 3 U, preferably 3 U to 6 U, per injection site and in a volume less than 0.030 ml.

4. The use according to any one of claims 1 to 3, wherein the botulinum toxin is injected at a dose of at least 3 U, preferably 3 U to 6 U, per injection site and in a volume of 0.010 ml to 0.025 ml.

5. The use according to any one of claims 1 to 4, wherein the botulinum toxin is injected into at least 3 injection sites, preferably 4 to 6 injection sites, more preferably 5 injection sites, in each platysma band.

6. The use according to any one of claims 1 to 5, wherein the botulinum toxin is injected into the at least one platysma band, and the total number of injection sites is 6 to 24, preferably 8 to 22, more preferably 14 to 20, and most preferably 16 to 20.

7. The use according to any one of claims 1 to 6, wherein the botulinum toxin is injected into the at least one platysma band, and the total volume is 0.15 ml to 0.60 ml, preferably 0.20 ml to 0.50 ml, more preferably 0.30 ml to 0.45 ml, and most preferably 0.35 ml to 0.4 ml.

8. The use according to any one of claims 1 to 7, wherein the botulinum toxin is injected into the at least one platysma band, and the total dose is 25 U to 200 U, preferably 30 U to 150 U or 40 U to 120 U, more preferably 50 U to 120 U or 60 U to 110 U, and most preferably 60 U to 100 U or 80 U to 100 U.

9. The use according to any one of claims 1 to 8, wherein the distance between injection sites along the platysma band is about 1 cm to about 3 cm, preferably about 1.5 cm to about 2.5 cm, and more preferably about 2 cm.

10. The use according to any one of claims 1 to 9, for neck and lower face rejuvenation, wherein the botulinum toxin is also injected into the mandibular contour of the subject.

11. The use according to claim 10, wherein the botulinum toxin is injected into the mandibular contour at a concentration of at least 50 U / ml, preferably at least 100 U / ml, and more preferably at least 150 U / ml, per injection site.

12. The use according to claim 10 or 11, wherein the botulinum toxin is injected into the mandibular contour at a dose of at least 1 U, preferably 1 U to 4 U, per injection site.

13. Use according to any one of claims 10 to 12, wherein the botulinum toxin is injected at at least 2 injection points on each mandibular contour, preferably 2 to 6 injection points on each mandibular contour, more preferably 3 to 5 injection points on each mandibular contour, and most preferably 4 injection points on each mandibular contour.

14. Use according to any one of claims 10 to 13, wherein the distance between the injection points along the mandibular contour is about 0.5 cm to about 2 cm, preferably about 1.0 cm to about 1.5 cm, and more preferably about 1 cm.

15. Use according to any one of claims 10 to 14, wherein the botulinum toxin is injected in each mandibular contour in a total volume of 0.02 ml to 0.16 ml and / or a total dose of 4 U to 16 U.

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