Injection of botulinum toxins into cervical latissimus for lower face and neck rejuvenation

Abstract

The present invention relates generally to the use of botulinum toxin for lower face and neck rejuvenation by injecting a high concentration of botulinum toxin into the latissimus laticus, more particularly into the latissimus laticus at the latissimus laticus belt and the mandibular contour. In particular, botulinum toxin is injected into the latissimus cervialis at the latissimus cervialis belt and the mandibular profile at a dose of 3 U to 5 U and a volume of 0.04 ml to 0.06 ml per injection site. Furthermore, the present invention relates to a method for rejuvenating the lower face and neck comprising injecting botulinum toxin into the latissimus at the latissimus belt and the mandibular profile at a dose of 3 U to 5 U and a volume of 0.04 ml to 0.06 ml per injection point.

Description

Technical Field

[0001] The present invention generally relates to the use of botulinum toxin for rejuvenation of the lower face and neck, by injecting a high concentration of botulinum toxin into the platysma muscle, more specifically into the platysma bands and the platysma muscle at the mandibular contour. Specifically, the botulinum toxin is injected into the platysma bands and the platysma muscle at the mandibular contour at a dose of 3U to 5U per injection site and a volume of 0.04 ml to 0.06 ml. In addition, the present invention relates to a method for rejuvenation of the lower face and neck, comprising injecting botulinum toxin into the platysma bands and the platysma muscle at the mandibular contour at a dose of 3U to 5U per injection site and a volume of 0.04 ml to 0.06 ml. Background Art

[0002] Restoring the volume of the mid-face and treating 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 thin 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 approximately 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 dilates the skin of the neck and pulls it downward. When viewed from the side, it acts in concert with the depressor anguli oris muscle (DAO) to lower the lower lip corner by pulling the outer corner of the mouth downward. 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 together 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] Various 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 muscular 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 botulinum toxin injection. 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 an active 150 kDa 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. Furthermore, the reduced downward pull of the injected platysma muscle 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 poses 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 low 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 low doses, the duration of action is shorter and more frequent treatments are required. Using low volumes may also result in fewer side effects due to reduced diffusion, but at the same time the toxin dose may be too low to produce an effective treatment.

[0009] To reduce unwanted side effects while achieving a high therapeutic effect, the impact of toxin dilution (concentration, volume) has been studied, but the results are controversial. On the one hand, it has been found that dilution increases the incidence of side effects observed in the treatment of glabellar wrinkles and 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). On the other hand, it has been found that a larger volume is beneficial for the treatment of dynamic forehead wrinkles, which requires fewer injections, thus reducing injection pain and increasing patient comfort and satisfaction (Hsu et al., Effect of volume and concentration on the diffusion of botulinum exotoxin A, Arch. Dermatol. 2004, 140(11): 1351 - 1354).

[0010] Regarding the 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., 2010, supra; 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 the platysma bands have been reported. OnabotulinumtoxinA with a total dose between 10 U and 100 U (see Brandt and Bellman, 1998, supra; Kane, 1999, supra; Brandt and Boker, 2003, supra; Tamura, 2012, supra) and up to 250 U (see Matarasso et al., 1999, supra) have been used for the successful treatment of the platysma bands. In addition, a total dose of 30 U to 60 U of ncobotulinumtoxin A (see Gubanova et al., 2013; supra; Prager et al., 2015, supra; also see the review by Sugrue et al., supra) has been described, and a total dose of 5 - 20 U per band of abobotulinumtoxinA (Dysport TM ) 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 U to 40+ U, depending on the number of injection sites (usually 3 to 5) and the dose used.

[0012] In addition, many different toxin dilutions are used in the art. For example, Matarasso et al., 1999 (supra) used 5 - 10 U of onabotulinumtoxin A in 0.1 - 0.2 mL per injection site Prager et al., 2015 (supra) injected 2.5 U of incobotulinumtoxin A in a volume of 0.0625 mL per injection site, and Gubanova et al., 2013 (supra), injected 5 U of incobotulinumtoxin A in a volume of 0.1 mL per injection site 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 per 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 platysma band treatment, overall neck tightening, mandibular contour reshaping (Brandt and Bellmann, 1998, see above; Tamura, 2012, see above) or lower face lift (Kane et al., 2010, see above) has been observed. Levy et al. focused specifically on 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 part of the platysma behind the nasolabial fold, along and below the mandible. Injecting botulinum toxin type A was found to release the tension on the mandibular contour by reducing the downward pressure of the platysma, thus making the elevator muscles dominant, resulting in skin lifting and mandibular contour reshaping.

[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 mandibular portion of the platysma described by Levy et al. (supra), but also treated the anterior fibers of the platysma and the mentalis muscle. This treatment pattern was reported to result in remodeling of the mandibular contour and improvement of the lateral buccal lines and the horizontal lines along the inferior border of the mandible. Bravo et al. (Bravo et al., J. Clin. Aesthet. Dermatol. 2019, 12(11):32-34) described another technique for improving mandibular contour and definition, including a first injection point 2-cm from the mandibular margin and two additional injection points anterior and superior to the first injection point.

[0015] Despite the development of the prior art, there remains a need for an improved rejuvenation treatment of the lower face and neck with botulinum toxin that produces highly satisfactory cosmetic results without endangering the safety of the patient.

[0016] Object of the present invention

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

[0018] The present invention is based on the discovery that administering a concentrated botulinum toxin preparation in a specific dose and volume to the platysma bands and the platysma at the mandibular contour can effectively and safely rejuvenate the lower face and neck. The specific dosing and injection regimens used according to the present invention target the toxin to the target platysma bands and the platysma at the mandibular contour, thereby providing the desired cosmetic effect while preventing the toxin from migrating to non-target muscles. A low migration potential is very important for avoiding spread to important anatomical structures in the neck, where muscles such as the swallowing muscles, the larynx, and the neck flexors are very close. 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 and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four injection points, preferably two or three injection points, more preferably two injection points, in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma bands, and at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma muscle at each mandibular contour.

[0020] Preferably, the dose injected at each injection point in the platysma bands and in the platysma muscle at (each) mandibular contour is 4 U to 5 U. The volume at each injection point in the platysma bands and in the platysma muscle at the mandibular contour is preferably 0.04 ml to 0.055 ml or 0.040 ml to 0.055 ml, more preferably 0.045 ml to 0.055 ml, most preferably 0.050 ml. This means that a high concentration of botulinum toxin solution is used in the present invention.

[0021] The high concentration (or high dose / low volume) administration mode is particularly suitable for relatively thin muscles such as the platysma muscle, not only the platysma muscle of the neck but also the platysma muscle in the mandibular contour area. Botulinum toxin injection in the mandibular contour improves the rejuvenation effect by relaxing the platysma muscle along the mandibular contour, thereby counteracting the downward pull of the platysma muscle. Therefore, the levator muscles will be dominant, resulting in an additional lifting effect and mandibular contour remodeling.

[0022] In a second aspect, the present invention relates to a method for neck and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four injection points, preferably two or three injection points, more preferably two injection points, in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma bands, and at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma muscle at each mandibular contour.

[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. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Describes the diffusion of toluidine dye in porcine muscle after intramuscular injection. (A): 10 μl, 20 μl, or 40 μl of toluidine dye was injected at a 45° injection angle; (B) 10 μl, 20 μl, or 40 μl of toluidine dye was injected at a 90° injection angle.

[0025] Figure 2 Shows an exemplary injection pattern according to the present invention for improving the prominence of the platysma band and the sharpness of the mandibular contour (left and right sides of the face with the same injection pattern). Detailed Description

[0026] The high-concentration (or high-dose / low-volume) method used in the present invention can not only safely administer botulinum toxin to the platysma band and the platysma muscle at the mandibular contour, but also produce excellent therapeutic effects and the desired rejuvenation effects on the lower face and neck that persist.

[0027] The relatively low volume is believed to prevent the toxin from diffusing to adjacent muscles, thereby preventing complications and unwanted side effects. This is particularly important in the case of the platysma muscle because the platysma muscle is a rather thin muscle and is anatomically close to important anatomical structures such as the swallowing muscles, the larynx, and the neck flexors. Therefore, special attention needs to be paid to preventing complications associated with the treatment of the platysma muscle, especially the platysma band, with botulinum toxin. The small volume used according to the present invention reduces the risk of migration / diffusion to unintended muscles, thereby eliminating the main concern in using botulinum toxin in cosmetic indications.

[0028] Although a smaller volume is injected according to the present invention, the dose administered at each injection site is still high due to the use of high-concentration botulinum toxin, 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 increasing patient satisfaction. In addition, the platysma muscle and the platysma band along the mandibular border can be treated simultaneously in one treatment session to achieve complete relaxation of the platysma muscle, thereby rejuvenating the lower face and neck. Treating the platysma muscle in the mandibular contour region not only results in rejuvenation of the lower face but also is expected to reduce the development of residual bands and compensatory bands.

[0029] In general, the present invention allows for treating the platysma as an anatomical and cosmetic entity and achieving a balanced and harmonious appearance of the lower face and neck, improving patient satisfaction without increasing side effects. Additionally, it enables the use of longer treatment intervals, resulting in a lower immunogenic risk of neutralizing antibody formation (known to cause secondary treatment failure). As a result, patient convenience and satisfaction are enhanced. Thus, low-volume and high-dose botulinum toxin treatment (i.e., using a high concentration 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 and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of a subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four (e.g., 2, 3, or 4) injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected at a dose of 3U to 5U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma bands, and at a dose of 3U to 5U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma muscle at each mandibular contour.

[0031] As used herein, the term "rejuvenation" broadly refers to restoring a youthful appearance. Additionally, 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 region from subnasale to menton, including the mandibular contour (including jowls and marionette lines). As used herein, the term "neck" refers to the region extending downward from the lower part of the mandible (lower jawbone) to the clavicle and sternum.

[0033] As used herein, the term "platysma" has the same meaning as the term "platysma muscle", and thus these terms may be 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 part of the pectoralis muscle, and a part of the platysma extends over the mandibular border 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 muscle at the mandibular contour. Since this part 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 muscle" 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 beside 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 injection into the platysma muscle, i.e., an intramuscular injection. Injecting botulinum toxin into the platysmal band can relax and flatten the platysmal band, making the band less noticeable. 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, the "platysmal band" is different from necklace lines, i.e., horizontal bands or lines that run horizontally around the front of the neck.

[0035] As used herein, the term "mandibular contour" refers to the lower edge of the lateral face below the cheek defined by the mandible. The singular use of "mandibular contour" refers to the mandibular contours on both sides (left and right) of the face, unless otherwise stated or the context otherwise indicates. Additionally, as described above, the platysma muscle passes through the mandible, with some fibers inserting into the bone below the oblique line and other fibers extending up to about 4 cm above the mandibular margin, entering the cheek, and inserting into the skin and subcutaneous tissue. Thus, as used herein, the term "injection into the mandibular contour" or "injection at the mandibular contour" or similar terms are intended to mean an injection into the platysma muscle, and more specifically, are intended to mean an intramuscular injection into the portion of the platysma muscle behind the nasolabial fold, along or under the mandible. Without being bound by theory, it is believed that an injection into the mandibular contour results in the release of tension on the mandibular contour by reducing the downward pressure of the platysma muscle. Thus, the elevator muscles are dominant, resulting in skin lifting and remodeling of the mandibular contour.

[0036] As used herein, the term "subject" is not particularly limited and refers to a person who requires cosmetic treatment of the neck, lower face, or both the neck and lower face. In the context of the present invention, the term "subject" can be used interchangeably with the term "patient". The age of the subject is preferably at least 30 years old, more preferably at least 40 years old or at least 50 years old, and most preferably at least 60 years old.

[0037] For the purposes of the present invention, a numerical value without a fractional digit should be understood as including all numerical values with one or more fractional digits, applying the general rounding rules to give the numerical value without a fractional digit. 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 the present application, if they do not have a fractional digit, can be a numerical value with a zero fractional digit, e.g., 4 can be 4.0.

[0038] As used herein, the term "botulinum toxin" is not particularly limited and includes botulinum toxins of any serotype (e.g., BoNT / A - H). For example, the botulinum toxin can 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 strain Hall. In addition, as used herein, the term "botulinum toxin" ("BT") and the synonymous term "botulinum neurotoxin" ("BoNT") refer to the pure botulinum neurotoxin and / or any complex thereof, i.e., any complex of the pure botulinum neurotoxin and a complexing protein (referred to as a "toxin complex"). Preferably, the botulinum toxin is a pure botulinum neurotoxin of serotype A.

[0039] As used herein, the term "pure botulinum neurotoxin" refers to a botulinum neurotoxin that does not contain a complexing protein (sometimes also referred to as the "neurotoxic component"), or more precisely, a botulinum neurotoxin that does not contain neurotoxin - associated proteins (NAPs). The pure botulinum neurotoxin is the (active) neurotoxic polypeptide that ultimately inhibits the release of acetylcholine. 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 can also be referred to herein as the "150 kDa neurotoxin", "botulinum neurotoxin (150 kD)" or "neurotoxic component". Preferably, the botulinum toxin is the (pure) botulinum neurotoxin contained in or is

[0040] As used herein, the term "toxin complex" refers to a high molecular weight complex of the neurotoxic component and a set of complexing proteins (NAP), such as the botulinum toxin type A complexes of 900 kDa, 500 kDa, and 300 kDa. The complexing proteins are non-toxic non-hemagglutinins (NTNHA), which are different hemagglutinins (HA) in serotype A - D strains. For example, the 900 kDa complex is contained in onabotulinumtoxin A ( Allergan, Inc., Irvine, CA, USA), and abobotulinumtoxin A ( Ipsen, Paris, France), (Ipsen / Galderma), and (Medytox) also contain the toxin complex as the active agent. Preferably, in addition to being contained in or being the pure botulinum neurotoxin, the botulinum toxin is the toxin complex contained in or or is or In another preferred embodiment, the botulinum toxin is not the toxin complex contained in or is not

[0041] The botulinum toxin can be a natural neurotoxin obtainable from 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 in 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. Botulinum toxins containing chemically modified amino acids are also included, such as one or more glycosylated, acetylated, or other modified amino acids. This may be beneficial for the uptake or stability of the toxin. Lipidation of the neurotoxic component is particularly preferred.

[0042] In the present invention, the dose is expressed in biological units because the botulinum toxin used can 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 (LD 50), i.e., the dose of toxin / neurotoxin that is capable of killing 50% of a group of mice. On this basis, as used herein, 1 unit (U) of toxin / neurotoxin is defined as one murine LD50 (1.0 LD50 = 1.0 U). The LD50 murine 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).

[0043] Another useful method for determining the biological activity (biopotency) of botulinum neurotoxin is the cell-based potency assay, which is 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 the murine intraperitoneal (i.p.) LD50 assay, since these values are calibrated using the LD50 reference standard.

[0044] 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. Therefore, within the framework of the present invention, the conversion rates provided below are used to establish incobotulinumtoxinA ("INCO"; botulinum toxin serotype A, without complexing proteins; 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 Type A; Mentor Worldwide LLC) relative potency. In this text, 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, in 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 using the above-mentioned MBA. Further 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-mentioned MBA.

[0045] In a preferred embodiment, a first aspect of the present invention relates to the use of botulinum toxin for neck and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma bands, and at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma muscle at each mandibular contour, 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 and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma bands, and at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma muscle at each mandibular contour, wherein 1 U corresponds to 1 LD50, and LD50 is the median lethal dose measured by 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 invention relates to the use of botulinum toxin for neck and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected into the platysma bands at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml, and into the platysma muscle at each mandibular contour at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml, where 1 U corresponds to 1 LD50, and LD50 is the median lethal dose measured by the mouse LD50 bioassay, provided that, if the botulinum toxin is incobotulinumtoxinA or onabotulinumtoxinA, 1 U of incobotulinumtoxinA and 1 U of onabotulinumtoxinA both equal 1 U, and if the botulinum toxin is abobotulinumtoxinA, 2.5 U of abobotulinumtoxinA equals 1 U.

[0046] 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 a process, composition, or formulation that comprises, includes, or contains an 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 a process, composition, or formulation. Further, 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 will be understood to refer to the exclusive inclusion of the indicated elements.

[0047] The terms "a", "an", and "the" and similar references used in the context of the present invention should 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.

[0048] According to the first aspect of the invention, the botulinum toxin is injected into the platysma bands at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml, and (independently) into the platysma muscle at each mandibular contour (i.e., the mandibular contours on the left and right sides of the subject's face) at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml.

[0049] Preferably, botulinum toxin is injected into the platysma band at a dose of 3.0 U to 5.0 U (e.g., 3.1 U, 3.5 U, 4.0 U, 4.5 U, and 4.9 U) per injection site, more preferably at a dose of 4 U to 5 U, particularly 4.0 U to 5.0 U (e.g., 4.1 U, 4.5 U, and 4.9 U) per injection site. Further preferably, botulinum toxin is injected into the platysma band at a volume of 0.045 ml to 0.06 ml (preferably 0.045 ml to 0.060 ml) or 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml) per injection site, more preferably at a volume of 0.045 ml to 0.055 ml per injection site, and most preferably at a volume of 0.05 ml per injection site.

[0050] Preferably, botulinum toxin is injected into the platysma at the mandibular contour on the left and right sides (i.e., both sides) of the subject's face at a dose of 3.0 U to 5.0 U (e.g., 3.1 U, 3.5 U, 4.0 U, 4.5 U, and 4.9 U) per injection site, more preferably at a dose of 4 U to 5 U, particularly 4.0 U to 5.0 U (e.g., 4.1 U, 4.5 U, and 4.9 U) per injection site. Further preferably, botulinum toxin is injected into the platysma at the two mandibular contours at a volume of 0.045 ml to 0.06 ml (preferably 0.045 ml to 0.060 ml) or 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml) per injection site, more preferably at a volume of 0.045 ml to 0.055 ml per injection site, and most preferably at a volume of 0.05 ml per injection site.

[0051] It is contemplated in the present invention that the dose and volume administered at each injection site in the platysma band may be the same or different from point to point, as long as the dose and volume at each injection site are within the indicated ranges (e.g., 3 U to 5 U and 0.04 ml to 0.06 ml). Similarly, the dose and volume administered at each injection site in the platysma at the mandibular contours on both sides of the subject's face may be the same or different from point to point, as long as the dose or volume at each injection site is within the indicated ranges (e.g., 3 U to 5 U and 0.04 ml to 0.06 ml). In addition, according to the present invention, the dose and volume injected into the platysma band are independently selected from the dose and volume injected into both the left and right mandibular contours.

[0052] In the present invention, the dose and volume for each injection site in the platysma band are preferably the same for all injection sites. Similarly, it is preferred that the dose and volume for each injection site in the platysma at the mandibular contour on both sides of the patient's face are the same for all injection sites. More preferably, the dose and volume for each injection site in the platysma band and in the platysma at the mandibular contour (on the left and right sides of the patient's face) are the same.

[0053] In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml).In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml). In a preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.06 ml (preferably 0.040 ml to 0.060 ml).

[0054] In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3U to 5U (preferably 3.0U to 5.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 3U to 5U (preferably 3.0U to 5.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3U to 4U (preferably 3.0U to 4.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 3U to 5U (preferably 3.0U to 5.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4U to 5U (preferably 4.0U to 5.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 3U to 5U (preferably 3.0U to 5.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3U to 5U (preferably 3.0U to 5.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 3U to 4U (preferably 3.0U to 4.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3U to 4U (preferably 3.0U to 4.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 3U to 4U (preferably 3.0U to 4.0U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml).In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.04 ml to 0.055 ml (preferably 0.040 ml to 0.055 ml).

[0055] In yet another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml).In yet another preferred embodiment, botulinum toxin is injected into the platysma bands at a dose of 3 U to 5 U (preferably 3.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma bands at a dose of 3 U to 4 U (preferably 3.0 U to 4.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml). In yet another preferred embodiment, botulinum toxin is injected into the platysma bands at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 4 U to 5 U (preferably 4.0 U to 5.0 U) per injection site and a volume of 0.045 ml to 0.055 ml (most preferably 0.050 ml).

[0056] In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 4 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 3 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 5 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 4 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 4 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 5 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma band at a dose of 5 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 3 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml).In another preferred embodiment, botulinum toxin is injected into the platysma bands at a dose of 5 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 4 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml). In another preferred embodiment, botulinum toxin is injected into the platysma bands at a dose of 5 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml), and into the platysma at each mandibular contour at a dose of 5 U per injection site and a volume of 0.045 ml to 0.055 ml (preferably 0.050 ml).

[0057] Furthermore, according to the present invention, botulinum toxin is preferably injected at a concentration of 50 U / ml to 125 U / ml, more preferably 60 U / ml to 120 U / ml, still more preferably 70 U / ml to 110 U / ml, and most preferably 80 U / ml to 100 U / ml. In particular, in the above "one preferred embodiment", "another preferred embodiment", and "yet another preferred embodiment", botulinum toxin is preferably injected at a concentration of 50 U / ml to 125 U / ml, more preferably 60 U / ml to 120 U / ml, still more preferably 70 U / ml to 110 U / ml, and most preferably 80 U / ml to 100 U / ml. A person skilled in the art can easily select a suitable concentration according to the dose to be injected, taking into account that for practical reasons and to avoid inaccurate dosing, the injection volume should not be too low.

[0058] Regarding the injection protocol, botulinum toxin is injected into two to four platysma bands. Preferably, botulinum toxin is injected into the two left and right medial bands or the two left and right lateral bands. More preferably, botulinum toxin is injected into each of the four platysma bands of the subject, namely the left medial band, the right medial band, the left lateral band, and the right lateral band.

[0059] Furthermore, in the context of the present invention, botulinum toxin is injected at four or five injection sites in each platysma band. Preferably, botulinum toxin is injected at five injection sites in each platysma band. More preferably, botulinum toxin is injected into two or four, preferably four, platysma bands, and at four or five injection sites in each platysma band. Most preferably, botulinum toxin is injected into four platysma bands, and at five injection sites in each platysma band.

[0060] In addition, botulinum toxin is injected at two to four injection sites, preferably three injection sites, and more preferably two injection sites in the platysma at each mandibular contour.

[0061] According to the present invention, it is particularly preferred to inject botulinum toxin into four platysma bands of a subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at two injection points in the platysma muscle at each mandibular contour and at five injection points in each platysma band. Further particularly preferably, the botulinum toxin is injected into four platysma bands of a subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at two injection points in the platysma muscle at each mandibular contour and at five injection points in each platysma band, wherein the botulinum toxin is injected at a dose of 3 U to 5 U (preferably 3.0 to 5.0 U) per injection point and in a volume of 0.04 ml to 0.06 ml (preferably 0.04 ml to 0.06 ml), or wherein the botulinum toxin is injected using any of the above dosing regimens.

[0062] The injection points along the platysma bands are preferably spaced at a distance of about 0.5 cm to about 2.0 cm, preferably about 0.5 cm to about 1.5 cm, more preferably about 1.0 cm. This spacing refers to the distance between adjacent injection points on the platysma band.

[0063] The injection points along each mandibular contour are preferably spaced at a distance of about 0.5 cm to about 3.0 cm or about 1.0 cm to about 3.0 cm, more preferably at a distance of about 1.0 cm to about 2.5 cm, still more preferably at a distance of about 1.5 cm to about 2.5 cm, most preferably at a distance of about 2.0 cm. Preferably, botulinum toxin is administered at two to four, preferably three, most preferably two injection points on each mandibular contour on each side of the patient's face, more specifically, generally along each mandible and below each jawbone (not before the line connecting from the nasolabial fold down to the mandible).

[0064] According to a preferred embodiment of the present invention, the injection pattern includes five injection points in each of the four platysma bands (a total of 20 injections) and two injection points in each mandibular contour on each side of the patient's face (a total of 4 injections), wherein the injection points along each platysma band are spaced at a distance of about 0.5 cm to about 2.0 cm, preferably about 0.5 cm to about 1.5 cm, more preferably about 1.0 cm and the injection points along each mandibular contour are spaced at a distance of about 1.5 cm to about 3.0 cm, preferably about 1.5 cm to about 2.5 cm, more preferably about 2.0 cm, or wherein the injection points along each platysma band are spaced at a distance of about 0.5 cm to about 1.5 cm, and the injection points along each mandibular contour are spaced at a distance of about 1.5 cm to about 2.5 cm.

[0065] For the use according to the first aspect of the invention, botulinum toxin can be administered in successive 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 one 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 successive administrations of botulinum neurotoxin can vary between about 5 and 10 months, particularly between about 6 and 9 months or between about 7 and 8 months. The longer duration of effect achieved by the present invention makes possible an extended treatment interval.

[0066] The extended duration of effect allows for a reduced frequency of administration of botulinum toxin. For example, botulinum neurotoxin can be repeatedly administered, wherein the time interval between the first and second administrations, or between the second or further administration 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.

[0067] Generally, the botulinum toxin used in the present invention is 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 formulation 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.

[0068] Specifically, the aqueous botulinum toxin solution or composition may contain other carriers or non-toxic, non-therapeutic, non-immunogenic stabilizers, etc. Thus, the aqueous botulinum toxin composition may 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 may be free of amino acids. In one embodiment, it may 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 may also contain non-ionic or ionic surfactants such as polysorbate or poloxamer. Suitable formulations for HSA-stabilized formulations containing botulinum toxin according to the present invention are disclosed, for example, in US 8,398,998 B2.

[0069] Preferably, the botulinum toxin used in the present invention is present 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 concentrations of sodium chloride, such as 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).

[0070] In a second aspect, the present invention relates to a method for rejuvenating the neck and lower face by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at three to four injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma bands, and at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml in the platysma muscle at each mandibular contour.

[0071] This treatment method is purely for aesthetic purposes, namely for the rejuvenation of the neck and 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. There is no particular limitation on the syringe, 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).

[0072] 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 preferred injection pattern is shown in which, according to this pattern, five injections are made at intervals of approximately 1 cm in the left and right medial bands and the left and right lateral bands, for a total of 20 injections into the platysma bands. In addition, two injections are made in the left mandibular contour and the right mandibular contour, with a distance of approximately 2 cm between the injection points, i.e., a total of four injections. According to this exemplary injection pattern, an improvement in the prominence of the platysma bands and the sharpness of the mandibular contour is obtained.

[0073] The method according to the second aspect of the present invention is closely related to the use according to the first aspect of the present invention. Therefore, all the definitions, explanations and advantages given herein regarding the use according to the first aspect also apply to the method according to the second aspect. In particular, the injection protocol and the administration protocol used in combination with the method according to the second aspect of the present invention are as disclosed above regarding the use according to the first aspect of the present invention.

[0074] Embodiment

[0075] Example 1

[0076] Intramuscular injection of dye into porcine muscle

[0077] 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.

[0078] Figure 1 Photographs of the muscle sections and a scale 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, which show the transverse dye diffusion in a top view.

[0079] From Figure 1It can be seen that diffusion depends on the injection volume and occurs along the muscle fibers (even with perpendicular injection). The lateral diffusion of 10 μl and 40 μl of toluidine dye solution at a 90° injection angle was 10.4 ± 2.2 mm and 23.3 ± 5.4 mm, respectively.

[0080] Although these results were obtained using porcine muscle in an ex vivo preclinical model, these results suggest that injecting high-concentration botulinum toxin at injection sites with a wide interval (e.g., 2 cm distance) in the platysma muscle provides sufficient distribution to achieve the desired therapeutic effect, even when only a small volume of botulinum toxin is injected.

[0081] Example 2

[0082] The injection protocol and dosing regimen of the present invention

[0083] A botulinum toxin solution is administered using the high-concentration (or "low-volume - high-dose") method according to the present invention. An exemplary dosing regimen according to the present invention is shown in Table 1.

[0084] Table 1. Dosing regimen

[0085]

[0086] 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 muscle (i.e., in the platysma muscle band and the platysma muscle at the mandibular contour).

[0087] Regarding the injection protocol, it is preferred to inject botulinum toxin using the injection protocol as Figure 2 shown. It can be seen that the injection protocol includes five injections at approximately 1 cm intervals in each of the four platysma muscle bands (the outer and inner bands on each side; a total of 20 injections in the platysma muscle bands), and two injections at approximately 2 cm intervals in the platysma muscle at the left and right mandibular contours (4 injections in the mandibular contours; a total of 24 injections in the platysma muscle bands and mandibular contours).

[0088] In the present invention, the injection protocol may also be modified to include only four injections in the four platysma bands, spaced approximately 1-2 cm apart (i.e., a total of 16 injections in the platysma bands), or more preferably, five injections in the left and right lateral bands, spaced approximately 1 cm apart, and four injections in the left and right medial bands (i.e., a total of 18 injections in the platysma bands), or four injections in the left and right lateral bands, spaced approximately 1-2 cm apart, and five injections in the left and right medial bands, spaced approximately 1 cm apart (i.e., a total of 18 injections in the platysma bands). Additionally, the above-modified injection protocols may be further modified to include three or four injections, spaced approximately 1-2 cm apart, preferably 1.5 cm apart, in the platysma of the left and right mandibular contours, instead of two injections (i.e., a total of 6 or 8 injections in the mandibular contours).

Claims

1. Use of botulinum toxin for neck and lower face rejuvenation by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of a subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected into four to five injection points in each platysma band and into two to four injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected into the platysma bands at a dose of 3U to 5U per injection point and a volume of 0.04 ml to 0.06 ml per injection point, and into the platysma muscle at each mandibular contour at a dose of 3U to 5U per injection point and a volume of 0.04 ml to 0.06 ml per injection point.

2. The use according to claim 1, wherein the botulinum toxin is injected into the platysma bands at a dose of 4U to 5U per injection point.

3. The use according to claim 1 or 2, wherein the botulinum toxin is injected into the platysma bands at a volume of 0.045 ml to 0.055 ml per injection point.

4. The use according to any one of claims 1 to 3, wherein the botulinum toxin is injected into the platysma muscle at each mandibular contour at a dose of 4U to 5U per injection point.

5. The use according to any one of claims 1 to 4, wherein the botulinum toxin is injected into the platysma muscle at each mandibular contour at a volume of 0.045 ml to 0.055 ml per injection point.

6. The use according to any one of claims 1 to 5, wherein the botulinum toxin is injected at a concentration of 50U / ml to 125U / ml.

7. The use according to claim 6, wherein the botulinum toxin is injected at a concentration of 60U / ml to 120U / ml or 70U / ml to 110U / ml.

8. The use according to any one of claims 1 to 7, wherein the botulinum toxin is injected into four platysma bands of the subject.

9. The use according to any one of claims 1 to 8, wherein the botulinum toxin is injected into five injection points in each platysma band.

10. The use according to any one of claims 1 to 9, wherein the botulinum toxin is injected into two injection points in the platysma muscle at each mandibular contour.

11. The use according to any one of claims 1 to 10, wherein the distance between the injection points along each platysma band is about 0.5 cm to about 2.0 cm.

12. The use according to claim 11, wherein the distance between the injection points along each platysma band is about 0.5 cm to about 1.5 cm.

13. The use according to any one of claims 1 to 12, wherein the distance between the injection points along the platysma muscle at each mandibular contour is about 0.5 cm to about 3.0 cm.

14. The use according to claim 13, wherein the distance between the injection points along the platysma muscle at each mandibular contour is about 1.5 cm to about 2.5 cm.

15. A method for rejuvenating the neck and lower face by injecting botulinum toxin into the platysma muscle, wherein the botulinum toxin is injected into two to four platysma bands of the subject and into the platysma muscle at the mandibular contour on both sides of the subject's face, wherein the botulinum toxin is injected at four to five injection points in each platysma band and at two to four injection points in the platysma muscle at each mandibular contour, and wherein the botulinum toxin is injected into the platysma bands at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml, and into the platysma muscle at each mandibular contour at a dose of 3 U to 5 U per injection point and a volume of 0.04 ml to 0.06 ml.

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