Use of botulinum toxin to reduce skin pore size and sebum production - Patents.com

Abstract

The present invention generally relates to the use of botulinum toxin to reduce pore size and / or sebum production in the skin using a specific dosing regimen and injection method. Specifically, botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 11 U / ml to 19 U / ml and a volume of 0.07 ml to 0.12 ml per injection site. The present invention further relates to a method for reducing pore size and / or sebum production in the skin using said specific dosing regimen and injection method.

Description

[Technical field]

[0001] The present invention generally relates to the use of botulinum toxin to reduce pore size and / or sebum production in skin using a specific dosing regimen and injection method. Specifically, botulinum toxin is administered by intradermal injection into a subject's skin at a concentration of 11 U / ml to 19 U / ml per injection site in a volume of 0.07 ml to 0.12 ml. Additionally, the present invention relates to a method for reducing pore size and / or sebum production in skin using said specific dosing regimen and injection method. [Background technology]

[0002] In recent years, holistic approaches using botulinum toxin have become increasingly popular, aiming not only to reduce wrinkles but also to improve skin quality. Also, due to the "selfie culture", there is a strong desire among people, especially young people, to have healthy looking skin. Good skin quality is perceived as healthy and undamaged skin, which is associated with a youthful and attractive appearance.

[0003] The primary mechanism of botulinum toxin is muscle paralysis by inhibiting the exocytosis of presynaptic acetylcholine, which is widely used in many cosmetic and therapeutic applications. More recently, intradermal injections of botulinum toxin have been shown to be effective in improving skin quality by reducing facial laxity, shortening mandibular length (resulting in lifted facial laxity), reducing sebum secretion, and reducing the number and density of facial pores. These effects may be due to paralyzing the arrector pili muscles (reducing pore size), inhibiting acetylcholine release (reducing sebum production), and affecting the superficial layers of the facial muscles (see, e.g., Park et al., Drugs Dermatol. 2021;20(1):49-54; Li et al., J. Dermatol. Sci. 2013;72:116-22).

[0004] Various techniques for intradermal injection of botulinum toxin are known in the art. One of them is the "mesobotox" or "microbotox" technique, in which highly diluted botulinum toxin (concentration less than 10 U / ml) is administered by injection. Small volumes (20 μL or less) are administered intradermally in at least 15 injections on the cheeks, at least 20 injections on the forehead, and at least 20-30 injections along the jawline / lower part of the face (Wu WTL. Facial rejuvenation without facelifts-personal strategies. Regional Conference, in Dermatol. Laser and Facial Cosm. Surg. 2002; September 13-15, 2002; paper presented at Hong Kong). In many cases, this results in more than 100 injection sites, which not only causes unnecessary pain for the patient, but also discomfort and time wasted for the injection provider.

[0005] Another technique described in the literature is the "SINB" technique (Superficial Injection Needling Botulinum) reported by Calvani et al. (Plast. Surg., 2019; 27(2): 156-161). It involves the injection of minute amounts of botulinum toxin into the skin using a needling technique consisting of multiple microdroplets with an electric device, rather than by a traditional syringe. The aim is to reduce sweat and sebaceous gland activity to improve skin texture. Compared to the previously described "mesobotox" or "microbotox" techniques, the resulting concentration is highly concentrated (>60 U / ml) rather than highly diluted, and therefore the reconstitution volume is different.

[0006] There are many publications regarding intradermal treatments that are aimed solely at improving enlarged pores and excessive sebum production, but the data on reconstitution volumes and dosages are different and even contradictory. Recommended reconstitution volumes range from 5 ml of saline per 100 U vial of botulinum toxin type A (BoNT / A) (resulting in a concentration of 20 U / ml) (see, e.g., Park et al., supra, and Shah, AR, J. Drugs Dermatol. 2008;7(9):847-850) to 3 ml per 300 U vial of BoNT / A (resulting in a concentration of 100 U / ml) (see, Rose and Goldberg (2012), Dermatol. Surg, 2012;39:443-448). Specifically, Park et al. (supra) administered 100 U of BoNT / A diluted in 5 mL of saline intradermally at 74 sites on the face (lateral face, anterior and medial cheeks, inferior margin of the mandible, and depressor anguli oris (DAO) muscle) using a 31G needle for a total dose of 92 U per treatment.

[0007] Min et al. (Aesthet. Surg. J., 2015;35(5):600-610) suggest using a smaller reconstitution volume of 2.5 ml per 100 U vial, resulting in a more concentrated solution containing 40 U / ml. Furthermore, Min et al. suggest using a larger injection volume (250 μL or even 500 μL per injection site) and distributing the injection sites over a standard 5 sites in the middle of the forehead. Other authors suggest injecting 100 μL of saline containing 2 U of BoNT / A per injection site, recommending injections 1 cm apart from each other (Shah, supra; Li et al., supra; Sayed et al., J. Dermatol. Treat. 2021,32(7):771-777).

[0008] Furthermore, WO 2004 / 016763A2 relates to the treatment of total glandular dysfunction with botulinum toxin, including reducing excess sebum production and pore size by intradermal injection of botulinum toxin. The specification discloses the treatment of excess sebum production by injecting BoNT / A and BoNT / B into the forehead, and BoNT / B into the skin of the brow ridge, forehead, and nose and nasolabial folds. However, the reconstitution volume, injection dose / volume, and injection mode for treating enlarged pores and excess sebum production are not clearly specified.

[0009] In the prior art, there are some drawbacks associated with treating enlarged skin pore size and excess sebum production. Using highly dilute approaches (higher reconstitution ratios are used, resulting in less concentrated solutions) results in less obvious and longer-lasting effects, resulting in more clinic visits for medical practitioners and less satisfaction for patients. On the other hand, using more concentrated doses does not seem to add efficacy (Shah, supra), but may require more injection sites, which may lead to more adverse events.

[0010] Furthermore, as described by Sapra et al. (J. Clin. Aesthet. Dermatol. 2017;10(2):34-44), undesirable effects may occur due to non-standardized injection schedules (i.e., patients are treated with treatment volumes based on individual needs that are not controlled, standardized, etc.) Thus, clear guidance is needed regarding reconstitution volumes, injection doses and volumes, and injection modes for treating enlarged skin pores and excess sebum production.

[0011] Furthermore, a major concern with the use of botulinum toxin A for cosmetic indications, apart from the lack of efficacy, which is always undesirable for both the injector and the patient, is the risk of migration / diffusion into unintended muscles. Thus, a dosing regimen and injection method that avoids unwanted diffusion of the toxin into the muscle layer located just beneath the treated dermis is highly desirable. Summary of the Invention [Problem to be solved by the invention]

[0012] Thus, there remains a need in the art for a treatment to reduce skin pore size and sebum production that overcomes the shortcomings of the prior art, is safe, and produces highly pleasing cosmetic results.

[0013] It is an object of the present invention to provide an improved botulinum toxin treatment that is effective and safe for reducing pore size and / or sebum production in the skin. [Means for solving the problem]

[0014] The present invention provides a specific dosing regimen and injection method for administering botulinum toxin to reduce skin pore size and / or sebum production. The proposed dosing regimen, optionally in combination with the proposed injection method, results in a balanced and harmonious appearance of the skin in the upper two-thirds of the face, and improves patient satisfaction without increasing side effects. More specifically, the relatively high dose provides an extended duration of effect, thereby improving patient convenience and satisfaction, and the relatively high volume provides the constant diffusion required to achieve the desired therapeutic outcome.

[0015] In a first aspect, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 11 U / ml to 19 U / ml in a volume of 0.07 ml to 0.12 ml per injection site.

[0016] Preferably, the botulinum toxin is administered at a concentration of 16 U / ml to 18 U / ml in a volume of 0.08 ml to 0.115 ml per injection site. The injection regimen preferably includes intradermal injection into 7 to 15 injection sites, preferably 11 to 13 injection sites, across the forehead including the glabellar region, and 6 to 12 injection sites, preferably 9 injection sites per cheek, with the distance between injection sites being 0.9 cm to 1.5 cm on the cheek and 1.7 cm to 2.3 cm on the forehead, the forehead including the glabellar region.

[0017] In a second aspect, the present invention relates to a method for reducing skin pores and / or sebum production, comprising administering botulinum toxin to the skin of a subject by intradermal injection at a concentration of 11 U / ml to 19 U / ml in a volume of 0.07 ml to 0.12 ml per injection site.

[0018] Preferred embodiments are set forth in the accompanying dependent claims and the following detailed description, taken in conjunction with the examples provided therein and the accompanying drawings. [Brief description of the drawings]

[0019] [Figure 1] 1 illustrates an exemplary injection pattern according to the present invention for reducing pore size and / or sebum production in the skin. [Diagram 2] The locations where sebum measurements were taken are shown: circles: sebumeter measurements; solid squares on the left: sebutape measurements; dashed squares on the right: spare sebutape measurements. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] The use of specific dosing schedules, optionally in conjunction with specific injection methods, according to the present invention allows for safe and effective treatment with reliable efficacy and predictable results.The safety of treatment is improved by reducing the diffusion of toxins to neighboring muscles, thereby reducing the possibility of associated adverse events in the face, such as asymmetrical smile, eyebrow and / or ptosis.At the same time, the relatively large volume administered still allows sufficient diffusion to achieve the desired treatment outcome.

[0021] In addition, the relatively high dose allows the duration of effect to be extended, thus improving patient convenience and satisfaction.In addition, the specific dosing schedule and injection method used herein, including reducing injection sites and increasing the volume per injection site, reduces injection pain, thus further improving patient satisfaction.Therefore, the present invention allows for an effective and safe treatment to reduce (enlarged) skin pore size and (excessive) sebum production, thereby making the skin of the upper two-thirds of the face look balanced and harmonious, improving patient satisfaction.

[0022] As used herein, the term "skin pore size" refers to the area of ​​a pore (i.e., the area occupied by a pore in a region of interest, e.g., in mm 2 The affected area is expressed in mm (e.g., mm2) and, since pores are three-dimensional, the volume of the pore (i.e., the total volume of the skin depression due to the presence of the pore in the selected area) is expressed in mm 3 In the context of the present invention, it is therefore the area affected by pores and the volume of the pores that is reduced. On the other hand, the term "pore count" or "pore number" refers to the number of individual pores detected within a predetermined range, expressed as a number. The term "skin pore size" in the phrase "use of botulinum toxin for reducing skin pore size" preferably relates to the size of enlarged skin pores.

[0023] "Enlarged skin pore size" within the meaning of the present invention can be defined as a skin pore volume that is at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, and most preferably at least 40% or at least 50% larger than the corresponding skin pore volume measured in a given area, in particular the cheek and forehead area, of a normal (average) subject. In the case of the cheek, the skin pore volume of a normal subject, measured for skin pores, i.e. detectable skin pores with a diameter between 0.1 mm and 1.0 mm, is greater than the corresponding skin pore volume of a normal subject in an area of ​​183 mm2 of the cheek. 2 1.25mm per (13.5mm x 13.5mm) 3 If we assume that, the size of the enlarged pore is 183 mm2 of cheek area. 2 (13.5mm x 13.5mm) ≧1.50mm 3 , preferably cheek area 183mm 2 Per ≧1.56mm 3 , and more preferably, a cheek area of ​​183 mm 2 Per ≧1.63mm 3 Or ≧1.69mm 3 , most preferably, a cheek area of ​​183 mm 2 Per ≧1.75mm 3 Or ≧1.88mm 3 It can be defined as:

[0024] For the measurement of skin pores, it is preferred to use a camera for image acquisition and corresponding software for skin analysis, in particular the Antera 3D imaging system (Miravex Limited, Dublin, Ireland). The term "Antera 3D imaging system" as used herein is used synonymously with the term "Antera 3D camera" and is intended to refer to the camera and analysis software system for skin analysis of Miravex Ltd. (Dublin, Ireland). It is possible to apply various filters to define the cutoff for detecting skin features such as the volume and area of ​​skin pores. The small filter detects pores with a diameter between 0.1 mm and 0.5 mm, the medium filter detects pores with a diameter between 0.1 mm and 1.0 mm, and the large filter detects pores with a diameter between 0.1 mm and 1.5 mm. Within the scope of the present invention, it is preferred to measure using an Antera 3D camera equipped with a medium filter.

[0025] For the forehead, the pore volume of normal subject skin is defined herein as 183 mm 2 1.15mm per (13.5mm x 13.5mm) 3 Therefore, it is assumed that the size of enlarged (i.e., at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, and most preferably at least 40% or at least 50% larger) skin pores, i.e., detectable skin pores with diameters between 0.1 mm and 1.0 mm, is greater than that of enlarged (i.e., at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, and most preferably at least 40% or at least 50% larger) skin pores measured in an area of ​​183 mm2 for the forehead. 2 (13.5mm x 13.5mm) ≧1.38mm 3 , preferably forehead area 183mm 2 Per ≧1.44mm 3 , more preferably forehead area 183 mm 2 Per ≧1.50mm 3 Or ≧1.55mm 3 , most preferably forehead area 183 mm 2 Per ≧1.61mm 3 Or ≧1.73mm 3 It can be defined as:

[0026] Alternatively or in addition to the above definition regarding the volume of skin pores in a given area, the term "enlarged skin pore size" within the meaning of the present invention can also be defined as an area affected by pores (area occupied by pores in a given area) that is at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, most preferably at least 40% or at least 50% larger than the corresponding area affected by pores measured in a given area, in particular in the cheek and forehead area, of a normal (average) subject, although this definition is less preferred. In the case of the cheek, the area affected by pores in a normal subject, measured for skin pores, i.e. detectable skin pores with a diameter between 0.1 mm and 1.0 mm, is at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, most preferably at least 40% or at least 50% larger than the corresponding area affected by pores measured in a given area, in particular in the cheek and forehead area, of a normal (average) subject. 2 (13.5mm x 13.5mm) 36.6mm 2 If we assume that the area affected by the enlarged pores is 183 mm2 of the cheek. 2 (13.5mm x 13.5mm) ≧43.9mm 2 , preferably cheek area 183mm 2 Per ≧45.8mm 2 , and more preferably, a cheek area of ​​183 mm 2 Per ≧47.6mm 2 Or ≧49.4mm 2 , most preferably, a cheek area of ​​183 mm 2 Per ≧51.2mm 2 Or ≧54.9mm 2 It can be defined as:

[0027] In the case of the forehead, the area affected by the pores in a normal subject in this specification is 183 mm 2 (13.5mm x 13.5mm) 33.7mm 2Therefore, it is assumed that the size of enlarged (i.e., at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, and most preferably at least 40% or at least 50% larger) skin pores, i.e., detectable skin pores with diameters between 0.1 mm and 1.0 mm, is greater than that of enlarged (i.e., at least 20%, preferably at least 25%, more preferably at least 30% or at least 35%, and most preferably at least 40% or at least 50% larger) skin pores measured in an area of ​​183 mm2 for the forehead. 2 (13.5mm x 13.5mm) ≧40.4mm 2 , preferably forehead area 183mm 2 Per ≧42.1mm 2 , more preferably forehead area 183 mm 2 Per ≧43.8mm 2 Or ≧45.5mm 2 , most preferably forehead area 183 mm 2 Per ≧47.2mm 2 Or ≧50.6mm 2 It can also be defined as:

[0028] The term "skin pores" or "pores" as used herein is intended to refer to depressions in the skin surface, typically having a rounded shape. Skin pores are relatively large in size and are visible to the naked eye. Skin pores within the meaning of the present invention can be defined as having a size in the range of 250 μm to 1000 μm or more, and thus have a surface area of ​​0.05 to 0.8 mm, assuming they have a rounded shape. 2 Macroscopically, they often look like small cuvettes, giving the skin microrelief a "golf ball" appearance. The "skin pores" according to the present invention are not openings or holes from eccrine sweat glands, but are much smaller in size.

[0029] As used herein, the term "sebum production" refers to the result of the production of a certain degree (or amount) of sebum (e.g., μg / cm) on the skin. 2"Sebum production" refers to the production of sebum, which results in the production of sebum (expressed in units of sebum per sebum). Preferably, the term "sebum production" as used herein refers to "facial sebum production". As used herein, the term "sebum" refers to an oily, waxy substance produced in the sebaceous glands, usually comprising triglycerides, wax esters, squalene, and other substances.

[0030] Sebum supports skin health in several ways, for example, it helps skin hydration and protects skin from potentially harmful pathogens such as bacteria and fungi. However, overproduction or excessive production of sebum can lead to oily skin. People with oily skin may notice that their pores look enlarged and their skin looks greasy or shiny. Therefore, the term "sebum production" used in the present specification within the phrase "to reduce the size of skin pores and / or sebum production" can refer to "excessive sebum production".

[0031] Preferably, the term "excessive sebum production" as used herein refers to a sebum level measured by a Sebumeter (e.g., Sebumeter® SM 815, Courage+Khazaka) at normal room conditions (e.g., 20°C, 40-60% relative humidity) of >180 μg / cm on the forehead, T-zone and scalp. 2 , >100 μg / cm in hair 2 , >150 μg / cm on cheeks, eyelids, and temples 2 and >130 μg / cm at the corners of the mouth. 2 When calculating the absolute amount of sebum on the entire forehead, this μg / cm 2 In terms of unit values, a typical frame is 6cm x 13cm, so 78cm 2 This can be calculated by multiplying

[0032] It should further be noted that the use of botulinum toxin to reduce sebum production according to the present invention is preferably for purely cosmetic (aesthetic) use, for example for the treatment of healthy subjects suffering from increased or excessive sebum production, which is cosmetically undesirable but not related to a medical condition or disease. In other words, the use of botulinum toxin to reduce sebum production is a cosmetic use to improve the skin quality of a ("healthy") subject, and not for the treatment of a skin disease. In particular, the use of botulinum toxin to "reduce sebum production" according to the present invention preferably excludes medical conditions or diseases in which sebum is involved, in particular acne, sebaceous gland hyperplasia, sebaceous cysts (single, multiple), sebaceous adenomas (sebaceous epithelioma, adenoma, cytoma) and seborrheic dermatitis.

[0033] The term "forehead" as used herein refers to the area of ​​the face whose lower border is represented by the supraorbital rim, whose upper border is the hairline, and whose lateral border is represented by the temporal ridge. Furthermore, the term "cheeks" as used herein preferably refers to the mid-face area including the anterior and medial portions of the cheeks (nasolabial and buccal cheek subunits).

[0034] The term "subject" as used herein refers to a human being, particularly but not limited to, who is in need of cosmetic treatment, particularly on the face, more particularly on the upper 2 / 3 of the face. The term "subject" in the context of the present invention can be used interchangeably with the term "patient". Preferably, the term "subject" as used herein relates to a subject with enlarged pore size and / or excessive sebum production, as defined herein. Particularly preferably, the subject in the context of the present invention is a human being with enlarged pore size, which corresponds to a score of at least 3 on the SASSQ (Scientific Assessment Scale of Skin Quality) by the investigator.

[0035] Within the meaning of the present invention, a number with no decimal places shall be understood to include any number with one or more decimal places, with the general rounding rules applied to make the number with no decimal places. For example, the number 3 includes 2.5 or 2.50 (rounded up to 3) and 3.4 or 3.49 (rounded down to 3) and all numbers therebetween. The same applies to the end points of the range. That is, the range 3-5 can mean, for example, 2.5-3.4 to 4.5-5.4. When one or both, preferably both, of each number or range end points disclosed herein does not have a decimal place, it is also intended that they can be (and can be replaced by) a number with a decimal place that is zero, for example, 4 can be 4.0.

[0036] The term "botulinum toxin" as used herein includes, but is not limited to, a botulinum toxin of any serotype (BoNT / A-H), preferably serotypes A and B, with or without a complex protein. Thus, within the scope of the present invention, the term "botulinum toxin" includes a toxin complex containing a 150 kDa botulinum neurotoxin and a complex protein (e.g., a 500-900 kDa botulinum toxin complex) or a 150 kDa neurotoxin without a complex protein. In the context of the present invention, the botulinum toxin is preferably a 150 kDa neurotoxin, particularly serotype A, without a complex protein.

[0037] The term "comprising", like the terms "including" and "containing" and any variations thereof, such as "comprises", "includes" and "contains", is intended to mean a non-exclusive inclusion such that a process, method, product-by-process, composition or formulation that comprises, includes or contains an element or list of elements does not necessarily include only those elements but may include other elements not expressly listed in such process, method, product-by-process, composition or formulation. In addition, within the framework of the present invention, the terms "comprise", "comprising", "includes", "including", "contains", "containing" and any variations thereof are intended to be replaced with the terms "consists" or "consisting" or any variations thereof, which are each to be understood to mean the exclusive inclusion of the specified elements. Furthermore, as used in the context of the present invention, the terms "a" and "an" and "the" and similar references are to be construed to cover both the singular and the plural, and therefore may also refer to "at least one" or "one or more," unless otherwise specified herein or clearly contradicted by context.

[0038] In a first aspect, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 11 U / ml to 19 U / ml in a volume of 0.07 ml to 0.12 ml per injection site.

[0039] Botulinum toxin also contains 12U / ml~19U / ml, 13U / ml~19U / ml, 14U / ml~19U / ml, 15U / ml~19U / ml, 16U / ml~19U / ml, 17U / ml~19U / ml, 18U / ml~19U / ml, 11U / ml~18 U / ml, 12U / ml~18U / ml, 13U / ml~18U / ml, 14U / ml~18U / ml, 15U / ml~18U / ml, 16U / ml~18U / ml, 17U / ml~18U / ml, 11U / ml~17U / ml, 12U / ml~17U / ml, 13 It can also be administered at a concentration of 1U / ml to 17U / ml, 14U / ml to 17U / ml, 15U / ml to 17U / ml, 16U / ml to 17U / ml, 11U / ml to 16U / ml, 12U / ml to 16U / ml, 13U / ml to 16U / ml, 14U / ml to 16U / ml, 15U / ml to 16U / ml, 11U / ml to 15U / ml, 12U / ml to 15U / ml, 13U / ml to 15U / ml, 14U / ml to 15U / ml, 11U / ml to 14U / ml, 12U / ml to 14U / ml, or 13U / ml to 14U / ml.

[0040] Additionally, the botulinum toxin can be administered in a volume of 0.08ml to 0.12ml, 0.09ml to 0.12ml, 0.10ml to 0.12ml, 0.11ml to 0.12ml, 0.07ml to 0.11ml, 0.08ml to 0.11ml, 0.09ml to 0.11ml, 0.10ml to 0.11ml, 0.07ml to 0.10ml, 0.08ml to 0.10ml, 0.09ml to 0.10ml, 0.07ml to 0.09ml, or 0.08ml to 0.09ml per injection site.

[0041] Additionally, the botulinum toxin can be administered at a dose of 1.4U to 1.9U, 1.5U to 1.9U, 1.6U to 1.9U, 1.7U to 1.9U, 1.8U to 1.9U, 1.4U to 1.8U, 1.5U to 1.8U, 1.6U to 1.8U, 1.7U to 1.8U, 1.4U to 1.7U, or 1.5U to 1.7U, 1.5U to 1.6U, or 1.6U to 1.7U per injection site.

[0042] It is contemplated that within the framework of the present invention, the concentration ranges of botulinum toxin disclosed herein can be combined with the volumes, doses, or volumes and doses of botulinum toxin disclosed herein.

[0043] Preferably, the botulinum toxin is administered by intradermal injection at a concentration of 15 U / ml to 19 U / ml in a volume of 0.075 ml to 0.12 ml per injection site, more preferably at a concentration of 16 U / ml to 19 U / ml in a volume of 0.075 ml to 0.115 ml per injection site, and most preferably at a concentration of 16 U / ml to 18 U / ml in a volume of 0.08 ml to 0.115 ml per injection site.

[0044] With respect to the injection regimen, the botulinum toxin is generally administered to the subject via intradermal injection across the forehead and / or cheeks, the forehead including the glabellar region. Preferably, the botulinum toxin is administered via intradermal injection to the forehead and cheeks, the forehead including the glabellar region, at a total dose of 30 U to 75 U, preferably 35 U to 65 U, more preferably 40 U to 60 U, and most preferably 45 U to 55 U.

[0045] The botulinum toxin is administered by intradermal injection into multiple injection sites, for example, 12-75 injection sites, preferably 15-60 injection sites, more preferably 20-50 injection sites, and most preferably 25-40 injection sites, distributed across the forehead and both cheeks, including the glabellar region. Particularly preferred within the scope of the present invention, the botulinum toxin is administered by intradermal injection into 7-15 injection sites, preferably 11-13 injection sites, distributed across the forehead and 6-12 injection sites per cheek, preferably 9 injection sites, including the glabellar region.

[0046] With regard to the injection pattern, the injection sites are preferably arranged such that the distance between the injection sites is 0.9 cm to 1.5 cm, more preferably 1.0 cm to 1.5 cm or 1.1 cm to 1.5 cm, most preferably 1.2 to 1.4 cm on the cheeks and 1.7 cm to 2.3 cm, preferably 1.8 cm to 2.2 cm, most preferably 2 cm on the forehead, the forehead including the glabella region. The term "distance" within the meaning of the present invention refers to the distance between any given injection site and another injection site closest to said given injection site, unless otherwise specified. It is also contemplated in the present invention that the distance between any two adjacent injection sites does not exceed 1.6 cm to 2.2 cm on the cheeks and 2.4 cm to 3.0 cm on the forehead.

[0047] Moreover, according to a particularly preferred injection pattern, botulinum toxin is administered by intradermal injection into 9 injection sites per cheek and 7-15 injection sites across the forehead, preferably 10-12 or 11-13 injection sites, where the forehead includes the glabellar region, and the 9 injection sites per cheek are arranged on three horizontal lines and three vertical lines, said lines intersecting each other to define said 9 injection sites. Additionally, separately or in conjunction with said particularly preferred cheek injection pattern, it is particularly preferred that 4 of the forehead injection sites are located on a first horizontal line, 5 of the forehead injection sites are located on a second horizontal line below the first horizontal line, and optionally 1 injection site is located in the glabellar region.

[0048] Particularly preferably, the botulinum toxin is administered by injection into twelve injection sites distributed over the forehead, including the glabellar area, four of the injection sites located on a first horizontal line, five of the injection sites located on a second horizontal line below the first horizontal line, two of the injection sites located above the left and right orbital rims (preferably at least 2 cm above the orbital rims) and below the four and five injection sites, and one of the injection sites located midway between the glabellar areas.

[0049] The term "botulinum toxin" as used herein is not particularly limited and includes any serotype of botulinum toxin (BoNT / AH). For example, the botulinum toxin may be serotype A or B (BoNT / A, BoNT / B). Preferably, the botulinum toxin is of serotype A, more preferably of serotype A1 (BoNT / A1), most preferably BoNT / A1 produced by the Clostridium botulinum Hall strain. Furthermore, the term "botulinum toxin" ("BT") as used herein and the interchangeable term "botulinum neurotoxin" ("BoNT") are intended to refer to pure botulinum neurotoxin and / or any complex thereof, i.e., any complex of pure botulinum neurotoxin and complex protein (referred to as "toxin complex"). Preferably, the botulinum toxin is a single serotype A botulinum neurotoxin.

[0050] As used herein, "botulinum neurotoxin alone" refers to botulinum neurotoxin without the complex proteins (sometimes referred to as "neurotoxin component"), more precisely, without the neurotoxin associated protein (NAP). Botulinum neurotoxin alone is the (active) neurotoxic polypeptide that ultimately inhibits the release of acetylcholine. It is a two-chain protein composed of a light chain (LC; about 50 kDa) and a heavy chain (HC; about 100 kDa) linked by disulfide bonds. Thus, the active neurotoxic polypeptide is sometimes referred to herein as "150 kDa neurotoxin", "Clostridium botulinum neurotoxin (150 kD)" or "neurotoxin component". Preferably, the botulinum toxin is the (alone) botulinum neurotoxin contained in Xeomin® or is Xeomin®.

[0051] As used herein, the term "toxin complex" refers to a high molecular weight complex of a neurotoxin component and a series of complex proteins (NAPs), including the 900 kDa, 500 kDa and 300 kDa C. botulinum type A toxin complexes. The complex proteins are the non-toxin non-hemagglutinin component (NTNHA) and the hemagglutinin component (HA) which differs among strains of serotypes A-D. For example, the 900 kDa complex is contained in onabotulinumtoxinA (Botox® / Vistabel®, Allergan, Inc., Irvine, Calif., USA), and abobotulinumtoxinA (Dysport®, Azzalure®, Ipsen, Paris, France), Alluzience® (Ipsen / Galderma), and Innotox® (Medytox) also contain a toxin complex as an active agent. Preferably, the botulinum toxin is a botulinum neurotoxin alone as contained in Zeomine® or Zeomine®, or a toxin complex contained in Botox® or Dysport®, or is Botox® or Dysport®.

[0052] The botulinum toxin may be a naturally occurring neurotoxin available from the bacterium Clostridium botulinum, or any other botulinum toxin, such as a botulinum toxin available from recombinant techniques and alternative sources, including genetic or chemical modifications. Chimeric or genetically modified botulinum toxins, i.e., botulinum toxins containing mutations such as substitutions, deletions and insertions, are also encompassed by the terms "botulinum toxin", "neurotoxin component" and similar terms. Preferably, the mutations do not impair any of the biological activity of the botulinum toxin. However, it is also contemplated to use mutations that modulate the biological activity of the botulinum toxin. Also encompassed are botulinum toxins that contain chemically modified amino acids, e.g., one or more amino acids that have been glycosylated, acetylated or otherwise modified that may be beneficial to the uptake or stability of the toxin. Lipid modifications of the neurotoxin component are particularly preferred.

[0053] Within the scope of the present invention, doses are expressed in biological units, since the botulinum toxins used may contain, for example, various proportions of inactive toxins that do not contribute to efficacy but contribute to the total protein load. In the context of the present invention, the biological potency of a botulinum toxin is determined by the Mouse Bioassay (MBA). The MBA allows the determination of the mean lethal dose (LD ) after intraperitoneal injection of the toxin / neurotoxin in mice. 50), i.e., the dose of toxin / neurotoxin that can kill 50% of the group of mice, is determined. On this basis, one unit (U) of toxin / neurotoxin as used herein is defined as the LD50 in mice (1.0 LD50=1.0U). The LD50 bioassay in mice is the gold standard for botulinum toxins in various biological, chemical or immunological detection methods and is known to those skilled in the art (see, for example, Pearce, LB; Borodic, GE; First, ER; MacCallum, RD Measurement of botulinum toxin activity: Evaluation of the lethality assay. Toxicol. Appl. Pharmacol. 1994, 128, 69-77).

[0054] Other useful methods for determining the biological activity (biological potency) of botulinum neurotoxins are cell-based potency assays, e.g. as disclosed in WO 2009 / 114748, WO 2013 / 049508 or WO 2014 / 207109. Activity results obtained from such cell-based assays correspond to activity values ​​obtained in LD50 assays in intraperitoneal mice, since these values ​​have been calibrated using the LD50 as a reference standard.

[0055] Because manufacturers of commercially available botulinum toxin preparations use different LD50 tests, the potency units listed for the botulinum toxin preparations sold by each manufacturer are proprietary and cannot be simply compared. Thus, within the framework of the present invention, incobotulinumtoxinA ("INCO"; Xeomine®, Bocouture®; botulinum toxin serotype A free of complex proteins; Merz Pharmaceuticals GmbH), onabotulinumtoxinA ("ONA"; Botox®, Vistabel®; botulinum toxin serotype A complex; Allergan Inc.), abobotulinumtoxinA ("ABO"; Dysport®, Azzalure®; botulinum toxin serotype A complex; Medicis Pharmaceutical Corp., Galderma Lab.), rimabotulinumtoxinB ("RIM"; Myobloc®, NeuroBloc®; botulinum toxin serotype B complex; Solstice Neurosciences The following conversion ratios are used to compare the potencies of ONA and INCO ("TBD"; botulinum toxin serotype A; Mentor Worldwide LLC): As used herein, the conversion ratio of ONA and INCO is 1:1; the conversion ratio of ONA / INCO:ABO is 1:2.5; the conversion ratio of ONA / INCO:RIM is 1:50 and the conversion ratio of ONA / INCO:TBD is 1:1. Furthermore, preferably, in the context of the present invention, 1 U of INCO (Xeomine®) and 1 U of onabotulinumtoxinA ("ONA"; Botox®) are considered to correspond to a LD50 in mice (1.0 LD50) measured as described above, i.e. 1 U.

[0056] Generally, the botulinum toxin used within the scope of the present invention is in the form of a liquid composition. This liquid composition can be formulated using various techniques known in the art depending on the desired application. It can be provided as a ready-to-use liquid preparation or in the form of a lyophilized powder that is typically reconstituted with saline before use. Preferably, the botulinum toxin used within the scope of the present invention is in the form of an aqueous solution, more preferably a saline solution or a physiological saline solution, most preferably a phosphate buffered saline solution. This aqueous solution can further include one or more pharma- ceutically acceptable substances. Suitable pharma-ceutically acceptable substances include those well known in the art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania.

[0057] In particular, the aqueous solution or composition of the botulinum toxin may contain other carriers or non-toxic, non-therapeutic, non-immunogenic stabilizers and the like. Thus, the aqueous composition of the botulinum toxin 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. In one embodiment, it may be free of amino acids. In one embodiment, it may be free of stabilizing peptides (e.g., consisting of 5-50 amino acids, 10-40 amino acids or 15-30 amino acids). In one embodiment, suitable non-protein stabilizers are those disclosed in WO 2005 / 007185 or WO 2006 / 020208. The botulinum toxin composition may also contain a non-ionic or ionic surfactant, 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 Pat. No. 8,398,998 B2.

[0058] Preferably, the botulinum toxin used within the scope of the present invention is in the form of an aqueous solution comprising sodium chloride (NaCl), more preferably in the form of a physiological saline solution (i.e. a solution containing sodium chloride at a physiological concentration, e.g. about 9 g / l NaCl), the aqueous botulinum toxin solution (i) is free of other excipients (except NaCl), (ii) is free of human serum albumin (HSA) and a sugar, in particular a monosaccharide or disaccharide, (iii) is free of human serum albumin (HSA) and lactose, (iv) is free of human serum albumin (HSA) and sucrose, (v) is free of monosaccharides and / or disaccharides (e.g. lactose and / or sucrose), (vi) is free of a buffering agent, (vii) is free of a single amino acid, (viii) is free of human serum albumin (HSA), sodium chloride and lactose, or is free of HSA, sodium chloride and sucrose, or (ix) is free of HSA and sodium chloride, or any combination of (i) to (ix).

[0059] In a preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 14 U / ml to 18 U / ml in a volume of 0.08 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 14 U / ml to 18 U / ml in a volume of 0.09 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 14 U / ml to 18 U / ml in a volume of 0.09 ml to 0.10 ml per injection site.

[0060] In a further preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 19 U / ml in a volume of 0.08 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 19 U / ml in a volume of 0.09 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 19 U / ml in a volume of 0.09 ml to 0.10 ml per injection site.

[0061] In yet another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 18 U / ml in a volume of 0.08 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 18 U / ml in a volume of 0.09 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 18 U / ml in a volume of 0.09 ml to 0.10 ml per injection site.

[0062] In yet another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 18 U / ml in a volume of 0.08 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 18 U / ml in a volume of 0.09 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 18 U / ml in a volume of 0.09 ml to 0.10 ml per injection site.

[0063] In yet another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 17 U / ml in a volume of 0.08 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 17 U / ml in a volume of 0.09 ml to 0.11 ml per injection site. In another preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 17 U / ml in a volume of 0.09 ml to 0.10 ml per injection site.

[0064] In a particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 14 U / ml to 18 U / ml, in a volume of 0.08 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 14 U / ml to 18 U / ml, in a volume of 0.09 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 14 U / ml to 18 U / ml in a volume of 0.09 ml to 0.10 ml per injection site at a dose of 1.4 U to 1.8 U, preferably 1.5 U to 1.8 U, more preferably 1.6 to 1.8 U.

[0065] In a further particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 19 U / ml, in a volume of 0.08 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 19 U / ml, in a volume of 0.09 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 19 U / ml in a volume of 0.09 ml to 0.10 ml per injection site at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U.

[0066] In yet another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 18 U / ml, in a volume of 0.08 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 18 U / ml, in a volume of 0.09 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 15 U / ml to 18 U / ml in a volume of 0.09 ml to 0.10 ml per injection site at a dose of 1.4 U to 1.8 U, preferably 1.5 U to 1.8 U, more preferably 1.6 to 1.8 U.

[0067] In yet another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 18 U / ml, in a volume of 0.08 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 18 U / ml, in a volume of 0.09 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 18 U / ml in a volume of 0.09 ml to 0.10 ml per injection site at a dose of 1.4 U to 1.8 U, preferably 1.5 U to 1.8 U, more preferably 1.6 to 1.8 U.

[0068] In yet another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 17 U / ml, in a volume of 0.08 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 17 U / ml, in a volume of 0.09 ml to 0.11 ml per injection site, at a dose of 1.4 U to 1.9 U, preferably 1.5 U to 1.9 U, more preferably 1.5 to 1.8 U. In another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16 U / ml to 17 U / ml in a volume of 0.09 ml to 0.10 ml per injection site at a dose of 1.4 U to 1.7 U, preferably 1.5 U to 1.7 U, more preferably 1.6 to 1.7 U.

[0069] In yet another particularly preferred embodiment, the present invention relates to the use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered by intradermal injection into the skin of a subject at a concentration of 16.67 U / ml, in a volume of 0.10 ml per injection site, at a dose of 1.67 U.

[0070] Further, it is contemplated herein, particularly with respect to the preferred and particularly preferred embodiments described in the preceding eleven paragraphs, that the botulinum toxin is preferably administered by intradermal injection to the subject's entire forehead and / or cheeks, preferably to the entire cheeks and forehead, the forehead including the glabellar area. Furthermore, when the botulinum toxin is administered by intradermal injection to (in) the entire cheeks and forehead (the forehead including the glabellar area), the botulinum toxin is administered in a total dose of preferably 30 U to 75 U or 35 U to 65 U, more preferably 40 U to 60 U, and most preferably 45 U to 55 U.

[0071] In a second aspect, the present invention relates to a method for reducing skin pores and / or sebum production, comprising administering botulinum toxin to the skin of a subject by intradermal injection at a concentration of 11 U / ml to 19 U / ml in a volume of 0.07 ml to 0.12 ml per injection site.

[0072] This treatment method may be used purely for cosmetic purposes, ie, to improve certain skin texture characteristics with the goal of enhancing the overall appearance of the skin.

[0073] The method according to the second aspect of the invention is closely related to the use according to the first aspect of the invention, and therefore all definitions, explanations, advantages etc. given herein with respect to the use according to the first aspect apply equally to the method according to the second aspect.

[0074] The method according to the second aspect of the invention is closely related to the use according to the first aspect of the invention, and therefore all definitions, explanations and advantages given herein for the use according to the first aspect apply equally to the method according to the second aspect. EXAMPLES

[0075] Example 1 Dosage regimens and injection methods according to the present invention A particularly preferred treatment regimen (i.e., dosing regimen and injection method) for reducing enlarged pore size and / or excess sebum production in the skin involves reconstituting a 100U vial (e.g., NT 201 (Xeomine®)) with 6ml of saline (0.9% w / v NaCl solution), resulting in a concentration of 16.67U / ml. Alternatively, a 50U vial can be reconstituted with 3ml of saline.

[0076] The reconstituted 16.67 U / ml botulinum toxin formulation is then injected intradermally into 30 injection sites distributed across the subject's forehead and both cheeks as follows: 12 injection sites on the forehead (100 μl / 1.67 U each) and 9 injection sites per cheek (18 total; 100 μl / 1.67 U each). The injection pattern is shown in FIG. 1.

[0077] As can be seen from this figure, the distance between injection sites is approximately 2 cm for the forehead and approximately 1-1.5 cm for the cheek area (depending on the facial features, surface area and size of the patient's head). Any suitable injection device can be used for administration of the botulinum toxin solution by intradermal injection, such as a 0.5-1.0 ml syringe fitted with a 30G-33G needle (e.g., a 32G or 33G needle).

[0078] The described combination of reconstitution volume, injection dose / volume, and injection methodology effectively treats enlarged pores and excess sebum production in the upper two-thirds of the face while improving patient satisfaction without increasing side effects.

[0079] Example 2 A study to investigate the efficacy and safety of botulinum toxin in treating enlarged pores and excess sebum production The following describes a typical study to investigate the efficacy and safety of botulinum toxin in treating enlarged pores and excess sebum production on the face. In total, for example, 30 subjects (male or female) are enrolled in a randomized, double-blind, placebo-controlled study. The main inclusion criteria are enlarged pores (e.g., investigator-reported SASSQ (Scientific Assessment Scale of Skin Quality) score of at least high, e.g., ≥ 3), and oily skin on the forehead (e.g., > 180 μg / cm as measured by sebumetry). 2 ) and age, for example, 18-50 or 21-39 years. In particular, patients who have received botulinum toxin treatment and other facial cosmetic treatments (e.g., laser or ultrasound treatments) within the past 12 months will not be included in the study.

[0080] A total of 30 subjects are randomized and randomly assigned to two different treatment groups, e.g., 2:1 (botulinum toxin group: placebo group). All subjects receive intradermal injections of botulinum toxin (e.g., NT 201, incobotulinumtoxinA) or placebo in saline (0.9% sodium chloride) solution in the upper face (forehead and cheek area), e.g., using the dosing schedule and injection method described in Example 1. The placebo is a placebo solution (0.9% sodium chloride) containing only the excipients of the saline solution of the botulinum toxin used (i.e., sucrose and human serum albumin in the case of NT 201).

[0081] All randomized subjects are then followed for a period of, for example, 90 days (day of injection is considered as day 1). To assess the reduction of pore size (e.g., pore volume in this study) and sebum production, efficacy variables are measured at baseline (day 1) and at regular follow-up visits, for example, at weeks 1, 4, 8, and 12. In addition, safety variables are used to evaluate safety, including adverse events (AEs), such as adverse events of special interest (AESIs) and serious adverse events (SAEs). Specific efficacy variables suitable for evaluating the efficacy of botulinum toxin to reduce pore size and sebum production are described below.

[0082] Pore ​​volume. Pore volume is the total volume of the skin depression caused by the presence of pores in the selected area in mm 3 The change in pore volume (primary outcome measure) from baseline to a specific time point, e.g., 4 weeks, is measured in the left and right cheek areas and forehead using an image capture camera and corresponding skin analysis software. In particular, an Antera 3D (Miravex Limited, Dublin, Ireland) camera and corresponding skin analysis software can be used for this purpose (see, e.g., Messaraa et al., Antera 3D capabilities for pore measurements, Skin Res. Technol. 2018;24(4):606-613). The camera is placed on the skin of the cheek and forehead area without pressing too hard, and measurements are taken in triplicate using a medium filter. The cheek is measured from the central 900 mm 2 (30×30mm 2 ) range is analyzed. The forehead is the entire area, that is, 3136 mm 2 (56×56mm 2 In addition to the pore volume, the following pore parameters can be evaluated and analyzed: total number of pores (the number of individual pores detected within a predefined range, expressed as a numerical value) and the area affected by pores (the area occupied by pores within the region of interest, expressed as mm 2 (expressed as

[0083] A 13.5 mm × 13.5 mm (183 mm) sample was taken of the central cheek of a subject with normal skin. 2 ) and the pore volume measured with the Antera 3D medium filter was approximately 1.25 mm 3 (and the total number of pores is about 54). The same area (183 mm 2 ) pore volume is 1.25 mm 3 Assuming that the value exceeds this by at least 20%, the size of the enlarged pore (pore volume) is, for example, 183 mm on the cheek. 2 Range ≧1.50mm 3 It can also be defined as follows. The cheek evaluation range defined above is 900 mm 2 Considering that the baseline pore volume in the cheek area of ​​subjects with enlarged pores was approximately 7.4 mm2, as measured with the Antera 3D medium filter. 3 It is estimated that.

[0084] Sebum production. The lipid layer on the skin surface is composed of epidermal lipids produced by the sebaceous glands. The amount of sebum on the skin surface is an important factor in the assessment of skin condition and gives information about the number and activity of sebaceous glands in the examined area. In this study, the sebum level on the forehead [μg / cm 2 The time course and change from baseline in sebumetry is measured using a Sebumeter (e.g., Sebumeter® SM 815, Courage+Khazaka). For this purpose, a measurement area on the midline of the forehead (see FIG. 2), e.g. 2 Sebum was collected from the skin surface using a translucent plastic strip and the value was expressed in μg / cm 2 The midline of the forehead is defined as approximately 2 cm above the subject's eyebrows. Excessive sebum production on the forehead ("oily skin") is defined as a value >180 μg / cm 2 It is defined as being.

[0085] Sebum Composition. Changes in sebum composition on the forehead are measured using mass spectrometry methods known in the art. The following types of lipids are analyzed: triglycerides, squalene, wax esters, free fatty acids, and cholesterol. To perform the sebum composition analysis, sebum samples are collected from the skin surface using sebum absorbent tape (SebuFix, Courage+Khazaka) from the left or right side of the forehead, defined as mid forehead above the midpupillary line. In case of any technical error in applying the tape to the forehead or transferring it to the container, rendering it invalid, an additional tape can be collected from the opposite side of the mid forehead above the midpupillary line (see Figure 2 for sebum measurement location). Sebu tape is applied to the forehead for 10 minutes and is removed approximately 4 cm from the forehead. 2 Sebum is collected from the area of ​​100 mm Hg. The sebum tape is transferred to a sterile container and optionally stored at least −20° C. (optimally −80° C.) prior to lipidomic analysis using mass spectroscopy.

[0086] Skin oiliness. In addition, the oiliness of the skin is determined using the SOS (oily skin scale). The SOS is a validated 6-point scale that correlates with Sebumeter measurements (Baumann et al., A Validated Questionnaire for Quantifying Skin Oiliness, J. Cosmet. Dermatol. Sci. Appl., 2014, 4, 78-84). The SOS is assessed by a questionnaire to accurately determine skin oiliness and can be used to screen and recruit patients for the study. The change in the SOS over time in this study will be evaluated by the subjects.

[0087] SASSQ by subjects and investigator. SASSQ (Scientific Assessment Scale for Skin Quality) is a visual, standardized, scientific assessment tool for measuring skin quality in humans. In this study, the SASSQ changes over time for pore size, wrinkles, skin roughness, erythema, and blemishes are assessed by subjects and investigators to determine the change from baseline. The five-level image of the scale indicates the visible severity of these changes in the aging process. The response options for this scale are 0 - no change, 1 - mild change, 2 - moderate change, 3 - severe change, and 4 - very severe change. SASSQ divides the face into 11 face-clusters (forehead, glabella, right eye, left eye, left cheek, nose, right cheek, perioral area, left jawline area, chin area, and right jawline area). The face-clusters used in this study can be, for example, forehead, glabella, left cheek, and right cheek.

[0088] GAIS by investigator and subject. The GAIS (global aesthetic improvement scale) is a balanced 7-point Likert scale (range -3 to +3) commonly used in aesthetic medicine to rate the overall aesthetic improvement of appearance after treatment. The overall assessment of aesthetic improvement in pore appearance (size and volume) and sebum production compared to the pre-injection state recorded on baseline photographs will be assessed by the investigator and subject individually using the GAIS, starting at, for example, week 1.

[0089] FACE-Q. The FACE-Q® is a patient-reported outcome (PRO) assessment that can be used to measure the patient-perspective of the outcome of facial aesthetic procedures and products. Each FACE-Q® scale consists of a series of items (or questions) that evaluate a concept of interest. Patients answer each item on a 4-point scale and are asked to answer with their own face / facial features in mind. Responses on the scale are summed and converted to a score from 0 to 100. For most FACE-Q® scales, higher scores indicate better outcomes. The following FACE-Q questionnaires may be used in the study: FACE-Q Skin Satisfaction (appearance of facial skin), FACE-Q Facial Appearance Satisfaction (overall facial appearance), FACE-Q Ageing Assessment (how respondents feel about the apparent age of their face), FACE-Q Ageing Assessment Visual Analogue Scale (rating of perceived age compared to chronological age), FACE-Q Appearance-related Psychological Distress (distress about appearance) and FACE-Q Impact of Treatment on Early Life (how respondents felt after undergoing cosmetic treatment).

[0090] The primary outcome measure was the change in pore size, more specifically pore volume, from baseline. Another important outcome measure was the reduction in sebum production compared to baseline. Statistical analysis of the data showed that pore volume and sebum production were significantly reduced.

Claims

1. 1. Use of a botulinum toxin for reducing pore size and / or sebum production in the skin, wherein the botulinum toxin is administered to the skin of a subject by intradermal injection at a concentration of 11 U / ml to 19 U / ml in a volume of 0.07 ml to 0.12 ml per injection site.

2. 2. The use of claim 1, wherein the botulinum toxin is administered at a concentration of 13 U / ml to 19 U / ml, 14 U / ml to 19 U / ml, 15 U / ml to 19 U / ml, 16 U / ml to 19 U / ml, 14 U / ml to 18 U / ml, 15 U / ml to 18 U / ml or 16 U / ml to 18 U / ml.

3. 3. The use according to claim 1 or 2, wherein the botulinum toxin is administered in a volume of 0.08ml to 0.12ml, 0.08ml to 0.11ml, 0.09ml to 0.12ml, 0.09ml to 0.11ml or 0.10ml to 0.11ml per injection site.

4. 4. The use according to any one of claims 1 to 3, wherein the botulinum toxin is administered in a dose of 1.4U to 1.9U, 1.5U to 1.9U, 1.6U to 1.9U, 1.5U to 1.8U or 1.6U to 1.8U per injection site.

5. The use according to claim 1 or 2, wherein the botulinum toxin is administered by intradermal injection at a concentration of 15 U / ml to 19 U / ml in a volume of 0.075 ml to 0.12 ml per injection site.

6. 6. The use according to any one of claims 1 to 5, wherein the botulinum toxin is administered by intradermal injection across the forehead and / or both cheeks of the subject, the forehead including the glabellar area.

7. 7. The use according to any one of claims 1 to 6, wherein the botulinum toxin is administered by intradermal injection to the forehead and both cheeks in a total dose of 30U to 75U, preferably 35U to 65U, more preferably 40U to 60U, most preferably 45U to 55U, wherein the forehead includes the glabellar area.

8. 8. The use according to any one of claims 1 to 7, wherein the botulinum toxin is administered by intradermal injection into 12 to 75 injection sites, preferably 15 to 60 injection sites, more preferably 20 to 50 injection sites, and most preferably 25 to 40 injection sites, distributed over the forehead and both cheeks, the forehead including the glabellar area.

9. 9. The use according to any one of claims 1 to 8, wherein the botulinum toxin is administered by intradermal injection into 7 to 15 injection sites, preferably 11 to 13 injection sites, across the forehead and 6 to 12 injection sites, preferably 9 injection sites per cheek, the forehead including the glabellar area.

10. 10. The use of claim 8 or 9, wherein the botulinum toxin is administered by intradermal injection into 9 injection sites per cheek and 7 to 15 injection sites across the forehead, preferably 11 to 13 injection sites, wherein the forehead includes the glabellar area, and wherein the 9 injection sites per cheek are arranged on three horizontal and three vertical lines, said lines intersecting each other to define the 9 injection sites.

11. 11. The use according to any one of claims 8 to 10, wherein four of the injection sites on the forehead are located on a first horizontal line, five of the injection sites on the forehead are located on a second horizontal line that is below the first horizontal line, and optionally one injection site is located in the glabellar area.

12. 12. The use of any one of claims 8 to 11, wherein the botulinum toxin is administered by injection into 12 injection sites distributed across the forehead, the forehead including the glabellar area, four of the injection sites are located on a first horizontal line, five of the injection sites are located on a second horizontal line below the first horizontal line, two of the injection sites are located above the left and right orbital rims and below the four and five injection sites, and one of the injection sites is located midway between the glabellar areas.

13. The use according to any one of claims 1 to 12, wherein the shortest distance between the injection sites is between 0.9cm and 1.5cm on the cheek and between 1.7cm and 2.3cm on the forehead, the forehead including the glabellar area.

14. The use according to any one of claims 1 to 12, wherein the botulinum toxin is in the form of a toxin complex of a 150 kDa botulinum neurotoxin and a complexing protein or a 150 kDa neurotoxin without a complexing protein, and / or the botulinum toxin is of serotype A or B.

15. 15. A method for reducing skin pores and / or sebum production, said method comprising administering botulinum toxin to said skin of a subject by intradermal injection at a concentration of 11 U / ml to 19 U / ml in a volume of 0.07 ml to 0.12 ml per injection site, preferably said method being defined as described in any one of claims 2 to 14.

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