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WO2025012403A1 - Traitement d'états d'hyperpigmentation par la toxine botulique - Google Patents

Traitement d'états d'hyperpigmentation par la toxine botulique Download PDF

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Publication number
WO2025012403A1
WO2025012403A1 PCT/EP2024/069733 EP2024069733W WO2025012403A1 WO 2025012403 A1 WO2025012403 A1 WO 2025012403A1 EP 2024069733 W EP2024069733 W EP 2024069733W WO 2025012403 A1 WO2025012403 A1 WO 2025012403A1
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WIPO (PCT)
Prior art keywords
botulinum toxin
injection
injection site
per injection
dose
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PCT/EP2024/069733
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English (en)
Inventor
Kay Marquardt
Thomas Hengl
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Merz Pharma GmbH and Co KGaA
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Merz Pharma GmbH and Co KGaA
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Priority to AU2024288687A priority Critical patent/AU2024288687A1/en
Publication of WO2025012403A1 publication Critical patent/WO2025012403A1/fr
Priority to IL324338A priority patent/IL324338A/en
Priority to MX2025013627A priority patent/MX2025013627A/es
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/4886—Metalloendopeptidases (3.4.24), e.g. collagenase
    • A61K38/4893—Botulinum neurotoxin (3.4.24.69)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders

Definitions

  • the present invention relates to the use of botulinum toxin for the treatment of hyperpigmentation conditions.
  • the use comprises the intradermal injection of botulinum toxin in the skin of a patient affected with the hyperpigmentation condition at a specified depth, volume and dose, thereby specifically targeting the epidermal and upper dermal layers.
  • the intradermal injection of botulinum toxin according to the present invention may be used in the treatment of melasma.
  • Hyperpigmentation is a common condition which occurs when patches of skin become darker in color than the surrounding skin due to the production of excess melanin.
  • Melanin is produced by specialized cells called melanocytes. These cells are located in the basal layer (stratum basale) of the epidermis, the deepest epidermal skin layer forming an interface (dermal-epidermal junction (DE J)) with the dermal layer below.
  • the synthesis of melanin begins with the amino acid tyrosine, which is converted into dihydroxyphenylalanine (DOPA) by the enzyme tyrosinase.
  • DOPA dihydroxyphenylalanine
  • DOPA is then converted into dopaquinone, which is the precursor for both eumelanin and pheomelanin, two types of melanin that contribute to skin color.
  • Eumelanin exhibits black color and is responsible for darker skin tones, while pheomelanin exhibits brown- red color and is responsible for lighter skin tones.
  • the relative amounts of eumelanin and pheomelanin produced by melanocytes determine skin color. Once melanin is produced, it is transferred from the melanocytes to neighboring skin cells called epidermal keratinocytes. Melanin is then distributed evenly throughout the skin, where it absorbs and scatters UV radiation from the sun, helping to protect the skin from damage.
  • hyperpigmentation varies widely depending on the underlying cause, geographic location, and other factors. For example, studies have shown that up to 90% of people may experience some form of hyperpigmentation during their lifetime, with varying degrees of seventy. This can include age spots, melasma, post-inflammatory hyperpigmentation, and other types of hyperpigmentation. For instance, melasma is estimated to affect up to 5-10% of people in the United States, with a higher prevalence among women, particularly those who are pregnant or taking oral contraceptives.
  • hyperpigmentation is not harmful to one’s health, it can be a significant cosmetic concern for many people and affect their self-esteem and quality of life.
  • treatment options including topical creams containing ingredients like hydroquinone, kojic acid and retinoids, chemical peels, laser therapy, cryotherapy, and other procedures.
  • these hyperpigmentation treatments may have several side effects, such as skin irritation, hypopigmentation, allergic reactions and sun sensitivity, and the effectiveness of these treatments is limited resulting in unsatisfactory outcomes.
  • botulinum toxin (botulinum neurotoxin or BoNT) has been suggested as an alternative treatment option for hyperpigmentation conditions.
  • Botulinum toxin is a neurotoxin from Clostridium botulinum which blocks the release of acetylcholine into the synaptic cleft, thereby preventing cholinergic neuromuscular innervation.
  • WO 2018/222652 A1 discloses methods for treating hyperpigmentation by administering a composition comprising a botulinum neurotoxin BoNT/DC.
  • BoNT/DC reduces melanin content in a human skin equivalent model and in human explant skin.
  • the examples further describe the intraepidermal treatment of solar lentigines by injecting 50 pl BoNT/DC toxin per injection point using a 30 G needle as well as the intraepidermal treatment of melasma by injecting 2 pl BoNT/DC toxin per injection point using a multineedle dermal injector system.
  • BoNT is locally administered by subcutaneous or intramuscular injection.
  • the treatment of hyperpigmentation requires more shallow injections, typically injection in the dermis (intradermal injections).
  • intradermal injections have been used in the past years in an increasing number of applications.
  • intradermal injections are more difficult to carry out than subcutaneous and intramuscular injection. This is especially true for the targeting of active substances to the epidermal and upper dermal layers of the skin. If a syringe with a normal injection needle is used for intradermal injection, this will give variable results in terms of application area and/or injection depths.
  • the object of the present invention is to provide an improved botulinum toxin treatment of hyperpigmentation conditions, which is efficacious, reliable and cost-effective.
  • the present invention is based on the finding that injection of low volumes of BoNT in the dermis in a depth-controlled manner allows to specifically address the layers of viable epidermis and upper dermal skin layers, more specifically the stratum basale and neighboring layers, in a safe, accurate and reliable manner. Since the addressed stratum basale contains target cells such as melanocytes, the BoNT injection of the present invention may be used to treat a variety of melanin-related afflictions, in particular hyperpigmentation conditions due to excess melanin production such as melasma. Also, the delivery of low volumes of BoNT specifically to viable epidermal and upper dermal layers allows for the use of reduced amounts of BoNT, resulting in significant cost savings.
  • the present invention relates to the use of botulinum toxin for the treatment of hyperpigmentation conditions, comprising injecting botulinum toxin in a patient’s skin that is affected from the hyperpigmentation condition, wherein the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the distance between injection sites is at least 5 mm, more preferably 5 mm to 20 mm, still more preferably 5 mm to 15 mm, yet still more preferably 10 mm to 20 mm, and most preferably 10 mm to 15 mm.
  • the hyperpigmentation condition to be treated is not particularly limited and may be a condition selected from the group consisting of melasma, freckles, solar lentigines, cafe-au-lait macules, post- inflammatory hyperpigmentation (PIH), moles and hyperpigmented nevi.
  • a particularly preferred condition to be treated is melasma.
  • the mentioned conditions are generally cosmetic in nature since they refer to aesthetic characteristics of the body that may cause a person to feel unhappy with their appearance but are non-threatening or do not cause substantial health problems that require therapeutic interventions.
  • the present invention relates to a method for the treatment of hyperpigmentation conditions, comprising injecting botulinum toxin in a patient’s skin that is affected from the hyperpigmentation condition, wherein the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the present invention relates to the use of botulinum toxin for the treatment of rosacea and a method for the treatment of rosacea, respectively.
  • the use and method comprise injecting botulinum toxin in a patient’s skin that is affected from rosacea, wherein the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is preferably of type A.
  • the botulinum toxin may be in a form that is free of complexing proteins or is in the form of a complex that contains complexing proteins.
  • the botulinum toxin is of type A and is in a form that is free of complexing proteins or, in an alternative preferred embodiment, is of type A and in the form of a complex that contains complexing proteins.
  • FIG. 2 shows the distribution of fluorescence labelled inactive BoNT in an ex vivo human skin model after injection using a 1.4 mm U-needle
  • E epidermis
  • D dermis
  • Scale bar 0.1 cm.
  • A picture showing an overlay of fluorescence microscopy (concentric structure of light to dark grey shades around injection site) and brightfield microscopy (grey).
  • B graph showing the BoNT distribution in terms of the BoNT signal (relative fluorescence intensity) as a function of skin depth.
  • A pictures showing an overlay of fluorescence microscopy (concentric structure of light to dark grey shades around injection site) and brightfield microscopy (grey).
  • B graphs showing the BoNT distribution in terms of the BoNT signal (relative fluorescence intensity) as a function of skin depth.
  • hyperpigmentation is intended to refer to skin conditions that occur as a result of increased production of melanin and is characterized by patches of skin that are darker in color than the surrounding skin.
  • epidermis has the meaning commonly used in the art, unless otherwise stated. It is the outermost of the three layers of the skin, the middle layer being the dermis and the innermost layer being the hypodermis.
  • the epidermis is composed of 4 or 5 layers, wherein these layers from outermost to innermost are: stratum corneum, stratum lucidum (only on the palms and soles), stratum granulosum, stratum spinosum, and stratum basale.
  • stratum basale contains keratinocytes, melanocytes and Merkel cells.
  • the thickness of the epidermis of human skin differs by region.
  • the stratum corneum generally has roughly a thickness of about 10 pm to about 40 pm, typically about 20 pm, and the viable epidermal layers may have a thickness of about 30 pm to about 150 pm, typically about 40 pm to about 100 pm.
  • viable epidermis refers to all epidermal layers including the stratum basale, except for the stratum corneum.
  • the thickness of the dermis varies by its location on the body. In the context of the present invention, the thickness is typically between 1500 pm and 4000 pm.
  • the dermis can be further differentiated between papillary dermis and reticular dermis.
  • the papillary dermis is the thin, upper layer that contains capillaries.
  • the thickness of the papillary dermis is typically about 300 pm to about 400 pm.
  • the reticular dermis is the lower layer that is much thicker than the papillary dermis and contains connective tissues and dense collagen bundles.
  • the term "dermal-epidermal junction”, “DEJ” or “dermoepidermal junction” refers to a structure at the interface between the epidermis and the dermis. It comprises a dense collagen network that creates undulations of the dermal-epidermal junction (DEJ).
  • the DEJ measures about 80 nm and assures several role within the skin, namely, ensuring a good cohesion between the epidermis and the dermis to maintain skin integrity, providing a physical boundary layer between fibroblasts and keratinocytes.
  • upper dermis or “upper dermal layer(s)”, as used herein, refer to the layers of the dermis that are closer to the epidermis than to the hypodermis. These layers usually include the papillary layer and a limited part of the reticular layer of the dermis.
  • injection site means the location where the injection device is placed on the skin and botulinum toxin is injected into the dermis via one or more needles.
  • the injection site is the site on the skin where the single needle enters the skin, and which may also be referred to herein as "injection point". If the injection device has multiple needles, i.e. , two, three, four or more, the injection site is the location on the skin where the multiple needles enter the skin.
  • the term "distance between injection sites”, as used herein, refers to the distance between an injection site and another adjacent injection site. If the injection device used for injection of botulinum toxin is a syringe with a single needle, the distance between injection sites is the distance between the site on the skin where the single needle enters the skin in a first injection and the site on the skin where the single needle enters the skin in a second adjacent injection. If the injection device has multiple needles, i.e., two, three, four or more, the distance between injection sites is the shortest distance between any one of the multiple needles of the injection device entering the skin in a first injection and any one of the multiple needles of the injection device entering the skin in a second adjacent injection.
  • injection depth refers to the depth of injection as measured from the skin surface to the end of needle tip entering the skin to reach the dermis.
  • volume per injection site and dose per injection site refer to the volume and toxin dose (in Units) of the botulinum toxin injected at one injection site. If an injection device is used that has multiple needles, i.e., two, three, four or more, the terms “volume per injection site” and “dose per injection site” refer to the volume and dose of botulinum toxin injected at one site through all needles, not only through a single needle of the multi-needle injection device.
  • injection device refers to any device, such as a syringe, which is suitable for injecting botulinum toxin into the dermis.
  • the injection device may be a conventional syringe equipped with a single hollow needle or, preferably, a microneedle device.
  • a "microneedle device”, as used herein, refers to device having multiple needles (also referred to as “microneedles"), e.g., two, three, four, five or more needles.
  • the needles are generally hollow and in fluid contact with a reservoir of the device filled with the botulinum toxin solution to be injected.
  • the needles may be made of different materials such as stainless steel or silicone.
  • a microneedle device that is suitable for use with the present invention is, for example, the MicronJetTM 600 silicon microneedle device available from NanoPass Technologies Ltd.
  • subject is not particularly limited but generally refers to a human receiving therapeutic or cosmetic treatment.
  • the subject is preferably a human receiving a cosmetic treatment.
  • subject may be interchangeably used herein with the term "patient”.
  • an effective amount means the amount of botulinum toxin which is generally sufficient to affect a desired change in the subject. For example, where the desired effect is a reduction in melanin, an effective amount of botulinum toxin is that amount which causes at least a substantial reduction of melanin, and without resulting in significant toxicity.
  • numerical values without decimal places shall be understood to include all numerical values with one or more decimal places that, applying common rules of rounding, give the numerical value without decimal places.
  • the numerical value 3 includes 2.5 or 2.50 (which are rounded up to 3) and 3.4 or 3.49 (which are rounded off to 3), and all numerical values in between.
  • endpoints of ranges the same applies, i.e. , a range of 3 to 5 may mean, e.g., 2.5-3.4 to 4.5-5.4.
  • the present invention relates to the use of botulinum toxin for the treatment of hyperpigmentation conditions, comprising injecting botulinum toxin in a patient’s skin that is affected from the hyperpigmentation condition, wherein the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the injection depth is 0.45 mm to 0.90 mm, more preferably 0.50 mm to 0.80 mm, still more preferably 0.55 mm to 0.70 mm, and most preferably 0.60 mm. Due to the administration by injection no botulinum toxin is applied onto the skin and the selected injection depth results in no or only minimal backflow of botulinum toxin from the injection channel. Further, in accordance with the present invention the injection depth is reached in a controlled and reliable manner. For example, at least 80%, preferably at least 85%, more preferably at least 90%, yet more preferably at least 95% of the injections in accordance with the present invention have an injection depth of X ⁇ 50 pm, with X being 450 pm to 950 pm.
  • At least 80%, more preferably at least 85%, still more preferably at least 90%, and yet more preferably at least 95% of the injections in accordance with the present invention have an injection depth of X ⁇ 50 pm, wherein X is 550 pm to 750 pm.
  • at least 80%, more preferably at least 85%, still more preferably at least 90%, and yet more preferably at least 95% of the injections in accordance with the present invention have an injection depth of X ⁇ 50 pm, wherein X is 600 pm to 650 pm.
  • the volume per injection site is preferably 15 pl to 30 pl or 10 pl to 25 pl, more preferably 15 pl to 25 pl, and most preferably 20 pl to 25 pl.
  • the dose per injection site is preferably 0.50 U to 10.0 U, more preferably 1 .0 U to 8.0 U, still more preferably 1 .5 U to 6 U, and yet more preferably 2.0 U to 4.0 U.
  • the distance between injection sites is preferably at least 5 mm and more preferably 5 mm to 20 mm, e.g., 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 11 mm, 12 mm, 13 mm, 14 mm, 15 mm, 16 mm, 17 mm, 18 mm, 19 mm, and 20 mm. Still more preferably, the distance between injection sites is 5 mm to 15 mm, yet still more preferably 10 mm to 20 mm, and most preferably 10 mm to 15 mm.
  • the injection density i.e. , the number of injections per area treated, is typically 1 to 16 injections per 4 cm 2 . Preferably, the number of injections per area treated is 1 .5 to 8 injections per 4 cm 2 , more preferably 2 to 4 injections per 4 cm 2 .
  • the hyperpigmentation conditions to be treated are preferably selected from melasma, freckles, solar lentigines, cafe-au- lait macules, post-inflammatory hyperpigmentation (PIH), moles and hyperpigmented nevi.
  • a preferred hyperpigmentation condition to be treated herein is melasma.
  • hyperpigmentation conditions are cosmetic conditions, i.e., conditions that affect the appearance of a subject rather than the subject’s health and does not require any therapeutic treatment.
  • cosmetic conditions refer to aesthetic characteristics of the body, they may cause a person to feel unhappy with their appearance and affect a person's self-esteem and quality of life.
  • individuals may choose to seek cosmetic treatments to improve their appearance and boost their confidence.
  • botulinum toxin for the treatment of hyperpigmentation conditions of the present invention is for purely cosmetic purposes, i.e., botulinum toxin is used for improving or enhancing physical appearance rather than for therapeutic reasons, i.e., to treat or prevent a medical condition.
  • melasma refers to an acquired hypermelanosis that occurs in sun-exposed areas. It is a type of hyperpigmentation that is characterized by dark patches on the face, particularly on the forehead, cheeks, upper lip, and occasionally on the neck, and rarely on the forearms. It is more common in women and is often associated with hormonal changes, such as those that occur during pregnancy or with the use of birth control pills.
  • Freckles also known as “ephelides”
  • epides refers to small, flat, dark (typically brown) spots that are often hereditary and typically appear on the face, chest, and arms. Freckles are more common in people with fair skin. They are caused by an increase in melanin production in response to sun exposure.
  • solar lentigines also known as “age spots” or “liver spots”
  • age spots or “liver spots”
  • brown spots that appear on the skin as a result of aging and exposure to the sun. They are most common in areas of the skin that are frequently exposed to the sun, such as the face, hands, and arms.
  • cafe-au-lait macules mean a type of skin pigmentation condition characterized by flat, light to dark brown spots on the skin that are oval or irregularly shaped. These macules can be present at birth or develop later in childhood.
  • PIH postinflam matory hyperpigmentation
  • PIH postinflam matory hyperpigmentation
  • skin has been irritated or injured. It is characterized by the appearance of darkened patches or spots on the skin that are typically brown, black, or red-brown in color. In response to the irritation or injury of the skin, the body produces excess melanin, which leads to the darkening of the affected area.
  • Common causes of PIH include acne, eczema, psoriasis, insect bites, burns, cuts, and surgical scars. While PIH is not a serious medical condition and does not pose any health risks, it is often of significant cosmetic concern for affected individuals.
  • moles refers to pigmented areas of the skin that are usually round or oval in shape and can vary in size, color, and texture. Moles are usually brown or black and can be flat or raised. Moles can appear at any age, but most people develop them during childhood or adolescence. Specific examples of moles include congenital moles and acquired moles (usually caused by sun exposure).
  • hyperpigmented nevi relates to pigmented areas of skin that can be present at birth or acquired later in life. Examples include congenital melanocytic nevus, acquired melanocytic nevus, halo nevus, spitz nevus, blue nevus, Becker's nevus, Hori's nevus.
  • the term "therapeutic disorder”, as used herein, refers to a disorder or disease that affects the subject’s health and which is treated by therapy.
  • the term “therapy” relates to the treatment of a disorder or disease to restore health by curing the disease or to a prophylactic treatment aimed at maintaining health by preventing ill effects that would otherwise arise or to the alleviation of the symptoms of pain and suffering.
  • the term “therapeutic” or “therapy” relates to restoring the human body from a pathological to its original (healthy) condition, or to prevent pathology, whereas cosmetic treatment takes as its starting point a normal state in general or a healthy in the narrower sense.
  • the botulinum toxin used within the present invention is not particularly limited and includes botulinum toxin of any serotype (BoNT/A-H) in a form that is free of complexing proteins or in the form of a complex that contains complexing proteins.
  • the botulinum toxin is of serotype A or B (BoNT/A, BoNT/B), with serotype A (BoNTA) being particularly preferred.
  • the botulinum toxin is of serotype A, still more preferably of serotype A1 (BoNT/A1 ), and most preferably BoNT/A1 produced by Clostridium botulinum Hall strain.
  • the botulinum toxin is of type A and in a form that is free of complexing proteins or is in the form of a complex that contains complexing proteins and, more preferably, is botulinum toxin of type A in a form that is free of complexing proteins.
  • BT botulinum toxin
  • BoNT botulinum neurotoxin
  • the (active) neurotoxic polypeptide that ultimately inhibits acetylcholine release also referred to herein as the "pure botulinum neurotoxin", “neurotoxic component”, “150 kDa neurotoxin” or “Clostridium botulinum neurotoxin (150 kD)" and a toxin form that is a complex containing complexing proteins (i.e., a complex of the neurotoxic component and complexing proteins).
  • the botulinum toxin complex is a high-molecular complex of the neurotoxic component and a set of complexing proteins (NAPs), including the 900 kDa, 500 kDa, and 300 kDa C. botulinum type A toxin complexes.
  • the complexing proteins are nontoxic nonhaemagglutinin (NTNHA) and, in strains of serotype A-D, different haemagglutinins (HAs).
  • NTNHA nontoxic nonhaemagglutinin
  • HAs haemagglutinins
  • the 900 kDa complex is included in onabotulinumtoxin A (Botox®A/istabel®, Allergan, Inc., Irvine, CA, USA), and also abobotulinumtoxin A (Dysport®, Azzalure®, Ipsen, Paris, France), Alluzience® (Ipsen/Galderma) and Innotox® (Medytox) contain a toxin complex as active agent.
  • the botulinum toxin is the pure botulinum neurotoxin that is contained in Xeomin® or is Xeomin®, or is the toxin complex contained in Botox® or Dysport® or is Botox® or Dysport®.
  • the botulinum toxin may be a natural neurotoxin obtainable from the bacteria Clostridium botulinum or any other botulinum toxin such as a botulinum toxin obtainable from alternative sources, including recombinant technologies and genetic or chemical modification. Chimeric or genetically modified botulinum toxins, i.e., botulinum toxins containing mutations including substitutions, deletions and insertions, are also encompassed by the terms "botulinum toxin", "neurotoxic component” and the like.
  • the mutation does not compromise any of the biological activities of botulinum toxin.
  • botulinum toxin containing chemically modified amino acids, for example one or more amino acids which are glycosylated, acetylated or otherwise modified, which may be beneficial to the uptake or stability of the toxin. Particularly preferred is the lipidation of the neurotoxic component.
  • the dose is expressed in biological units because the used botulinum toxin may contain, for example, variable percentages of inactive toxin that contribute to the overall protein load without contributing to efficacy.
  • the biological potency of botulinum toxin is determined using the mouse bioassay (MBA).
  • the MBA determines the mean lethal dose (LD50) of toxin/neurotoxin after intraperitoneal injection in mice, i.e. , the dose of toxin/neurotoxin capable of killing 50% of a group of mice.
  • the LD50 mouse bioassay is the gold standard among various biological, chemical or immunological detection and activity determination methods 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).
  • Another useful method for determining the biological activity (biological potency) of a botulinum neurotoxin is a cell-based potency assay as it is disclosed, for example, in W02009/114748, WO 2013/049508 or WO 2014/207109.
  • the activity results obtained with such cell-based assays correspond to the activity values obtained in the mouse i.p. LD50 assay because the values are calibrated using the LD50 reference standard.
  • 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.
  • 1 U of INCO (Xeomin®) and 1 U of onabotulinumtoxinA (“ONA"; Botox®) shall be deemed to correspond to one mouse LD50 (1.0 LD50), or 1 U, measured as described above.
  • the botulinum toxin used within the present invention is in the form of a liquid composition.
  • the liquid composition can be formulated by various techniques dependent on the desired application, as known in the art. It may be provided as a ready-to-use liquid formulation or in the form of a lyophilized powder that is to be reconstituted, typically in physiological saline, prior to use.
  • the botulinum toxin used within 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 physiological saline solution.
  • the aqueous solution may additionally comprise one or more pharmaceutically acceptable substances. Suitable pharmaceutically acceptable substances comprise those well known in the art, see, e.g., Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania.
  • the aqueous botulinum toxin solution or composition may include other carriers or non-toxic, non-therapeutic, non-immunogenic stabilizers and the like.
  • the aqueous botulinum toxin composition may contain glycerol, protein stabilizers such as human serum albumin (HSA) or non-protein stabilizers such as polyvinyl pyrrolidone (PVP), hyaluronic acid or free amino acids, e.g., methionine or histidine.
  • HSA human serum albumin
  • PVP polyvinyl pyrrolidone
  • hyaluronic acid hyaluronic acid
  • free amino acids e.g., methionine or histidine.
  • Suitable non-proteinaceous stabilizers are disclosed in WO 2005/007185 or WO 2006/020208.
  • the botulinum toxin composition can also include non-ionic or ionic surfactant, e.g., polysorbate or poloxamer.
  • a suitable formulation for HSA-stabilized formulation comprising a botulinum toxin according to the present invention is, for example, disclosed in US 8,398,998 B2.
  • the botulinum toxin used within the present invention is in the form of an aqueous solution comprising sodium chloride (NaCI), preferably in the form of a physiological saline solution (i.e.
  • a solution including sodium chloride in physiological concentration e.g., about 9 g/l NaCI
  • a solution including sodium chloride in physiological concentration which further comprises one or more of the following (i) to (ix): (i) no other excipient (except NaCI), (ii) human serum albumin (HSA) and a sugar, in particular a monosaccharide or a disaccharide, (iii) human serum albumin (HSA) and lactose, (iv) human serum albumin (HSA) and sucrose, (v) a monosaccharide and/or a disaccharide (e.g.
  • the botulinum toxin is in the form of an aqueous formulation comprising botulinum toxin, sodium chloride and human serum albumin, or an aqueous formulation comprising botulinum toxin, sodium chloride, human serum albumin and lactose, or an aqueous formulation comprising botulinum toxin, sodium chloride, human serum albumin and sucrose.
  • Another particularly preferred aqueous formulation comprises botulinum toxin, sodium chloride, human serum albumin and histidine.
  • the botulinum toxin can be as defined herein above.
  • the present invention relates to a method for the treatment of hyperpigmentation disorders, comprising injecting botulinum toxin in a patient’s skin that is affected from the hyperpigmentation condition, wherein the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the hyperpigmentation conditions are not particularly limited and include the hyperpigmentation conditions as defined above.
  • 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.
  • all definitions, explanations and advantages given herein with respect to the use according to the first aspect equally apply to the method according to the second aspect.
  • the injection scheme and dosing regimen used in connection with the method according to the second aspect of the present invention is as disclosed herein above in relation to the use according to the first aspect of the present invention.
  • the botulinum toxin is preferably of type A.
  • the botulinum toxin may be in a form that is free of complexing proteins or is in the form of a complex that contains complexing proteins.
  • the botulinum toxin is of type A and is in a form that is free of complexing proteins or, in an alternative preferred embodiment, is of type A and in the form of a complex that contains complexing proteins.
  • the present invention relates to the use of botulinum toxin for the treatment of rosacea and a method for the treatment of rosacea, respectively.
  • the use and method comprise injecting botulinum toxin in a patient’s skin that is affected from rosacea, wherein the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • rosacea relates to a chronic skin condition that causes redness, flushing, and visible blood vessels in the face, especially the cheeks, chin, nose and mid forehead. It is not a life-threatening condition but of cosmetic concern and can cause enormous social and psychological problems. In other words, rosacea is considered to be a cosmetic disorder or state, rather than a therapeutic disorder. The mechanism of action of botulinum toxin in the treatment of rosacea is not yet fully understood.
  • Rosacea is considered to be a cosmetic condition, i.e. , a condition that affects the appearance of a subject rather than the subject’s health, as explained above in relation to the hyperpigmentation conditions.
  • the use and method according to the third and fourth aspects of the present invention are closely related to the use according to the first aspect of the present invention and the method according to the second aspect of the present invention, respectively, and differ from these only in that the condition to be treated is rosacea rather than a hyperpigmentation disorder.
  • all definitions, explanations and advantages given herein with respect to the use according to the first and second aspects equally apply to the use according to the third aspect and the method of the fourth aspect of the present invention.
  • any disclosure regarding the injection scheme and dosing regimen set out above in relation to the first and second aspect of the present invention is to be understood to also represent a disclosure for the third and fourth aspect of the present invention.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 15 pl to 25 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 ,l to 25
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, and at a dose of 0.25 U to 12.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 15 pl to 25 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 jil to 25 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, and at a dose of 0.50 U to 10.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 1.0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, and at a dose of 1.0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, and at a dose of 1 .0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, and at a dose of 1.0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 25 pl per injection site, and at a dose of 1 .0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, and at a dose of 1.0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, and at a dose of 1 .0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, and at a dose of 1 .0 U to 6.0 U per injection site.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 0.25 U to 12.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 0.50 U to 10.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 1 .0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 1 .0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 1.0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 1.0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 1.0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 1.0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 1.0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 1.0 U to 6.0 U per injection site, and at an injection distance of 5 mm to 20 mm, preferably 10 mm to 20 mm, and more preferably 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 0.25 U to 1.0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 0.25 U to 1 .0 U per injection site, and at an injection distance of 5 mm to 10 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 1.0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 1 .0 U to 6.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 1 .0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 1 .0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 1 .0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 1.0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 1 .0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 1 .0 U to 8.0 U, preferably 2.0 U to 7.0 U, more preferably 3.0 U to 6.0 U, per injection site, and at an injection distance of 10 mm to 20 mm or 10 mm to 15 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 10 pl to 30 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 30 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1.00 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 15 pl to 25 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.40 mm to 1 .00 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the botulinum toxin is intradermally injected at an injection depth of 0.50 mm to 0.80 mm, at a volume of 20 pl to 25 pl per injection site, at a dose of 8.0 U to 12.0 U per injection site, and at an injection distance of 15 mm to 20 mm.
  • the distribution of fluorescence labelled inactive BoNT was determined using an ex vivo human skin model.
  • the inactive BoNT (triple mutation in amino acid sequence of the light chain) was labeled with Alexa Fluor 647 and solved in PBS puffer (pH 7.4) at a concentration of 0.58 mg/mL.
  • the skin model consisted of ex vivo abdominal human skin. The hypodermis of the skin was nearly completely removed. The trimmed skin was clamped under tension and incubated at 37°C. The model was kept hydrated by a cloth soaked in PBS puffer (pH 7.4), which was in contact to the reticular dermis.
  • the MicronJet600 resulted in a BoNT concentration halo of around the depth of 1.0 mm, with a maximum BoNT concentration at a shallow injection depth.
  • the U-needle shows the highest BoNT concentration in a depth of around 1 .4 mm (see FIG. 2) and addresses therefore the mid-dermal region.
  • the standard G30/ needle resulted in a low injection accuracy with random injection depths (see FIG. 3; two exemplary injections are shown). In other words, the injections using a standard G30/ needle address the whole dermal layer randomly.
  • the Micron Jet600 injection bolus is surprisingly redirected to the upper skin layers, i.e. , the upper dermal region including the papillary region (thin top layer of the dermis) as well as the stratum basale (the deepest epidermal layer).
  • the target cells e.g., the melanocytes involved in pigmentation, which are located in the dermoepidermal junction (the interface between the basal epidermis and the papillary dermis).
  • the BoNT distribution was consistent in all injections with MicronJet600, thereby providing high injection accuracy and reliability.
  • the specific Micronjet targeting effect is limited to a lower injection volume of, e.g., about 0.05 mL. More specifically, a blue dye Toluidine blue was used as a surrogate for a BoNT solution and injected at different volumes into the dermis of the previously described ex vivo human skin model using the MicronJet600. Immediately after injection, the vertical spread of the dye was then determined by gross cryo-sectioning of the human skin model. Using an injection volume of 0.0125 ml, the maximal vertical spread was found to be 1 .5 mm. With a volume of 0.025 ml, the maximal vertical spread increased to 1.9 mm.
  • a blue dye Toluidine blue was used as a surrogate for a BoNT solution and injected at different volumes into the dermis of the previously described ex vivo human skin model using the MicronJet600.
  • the vertical spread of the dye was then determined by gross cryo-sectioning of the human skin model. Using an injection volume of 0.0125 m
  • the observed maximal vertical spread was 2.0 mm, which means that the whole dermis (max. dermal vertical dimension/length was in the particular case around 2 mm) is filled with the solution from the upper dermis to the lower dermis.
  • the botulinum toxin solution is administered using the exemplary dosing regimens according to the present invention as shown in Table 1. Table 1. Dosing regimens
  • the injection dose increases with the injection distance. For example, if the dose per injection point is 0.25 U to 2 U, the injection distance may be for example 5 mm. If the dose per injection point is 2 U to 8 U, the injection distance may be for example 10 mm, and if the dose per injection point is 4 U to 16 U, the injection distance may be for example 20 mm.
  • the injected botulinum toxin is typically a 0.9% saline solution.
  • a microneedle device is used to deliver the botulinum toxin to the upper dermis.
  • a syringe equipped with a Micron Jet600 microneedle device can be used. This device can be used for multiple injections without the need of exchanging the device after each injection.
  • the botulinum toxin is preferably injected using an injection scheme that includes multiple injections.
  • the number of injections depends on the size of the area to be treated (e.g., melasma area).
  • the injections are arranged in a grid-like pattern, e.g., in a square grid.
  • the lateral distribution of BoNT is about 5.4 ⁇ 0.3 mm after 4 hours. Therefore, depending on the dose per injection site, a suitable distance between injection sites is about 5 mm or higher.

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Abstract

La présente invention concerne l'utilisation de toxine botulique pour le traitement d'états d'hyperpigmentation. L'utilisation comprend l'injection intradermique de toxine botulique dans la peau d'un patient atteint de l'état d'hyperpigmentation à une profondeur, un volume et une dose spécifiés, ciblant ainsi spécifiquement les couches dermiques et épidermiques supérieures. Par exemple, l'injection intradermique de toxine botulique selon la présente invention peut être utilisée dans le traitement du mélasma.
PCT/EP2024/069733 2023-07-12 2024-07-11 Traitement d'états d'hyperpigmentation par la toxine botulique Pending WO2025012403A1 (fr)

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AU2024288687A AU2024288687A1 (en) 2023-07-12 2024-07-11 Treatment of hyperpigmentation conditions by botulinum toxin
IL324338A IL324338A (en) 2023-07-12 2025-10-30 Treatment of hyperpigmentation conditions using botulinum toxin
MX2025013627A MX2025013627A (es) 2023-07-12 2025-11-13 Tratamiento de condiciones de hiperpigmentacion mediante toxina botulinica

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EP23185114.8 2023-07-12

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005007185A2 (fr) 2003-07-22 2005-01-27 Biotecon Therapeutics Gmbh Formulation destinee a un agent pharmaceutique proteique ne contenant pas d'albumine serique humaine ajoutee (hsa)
WO2006020208A2 (fr) 2004-07-26 2006-02-23 Merz Pharma Gmbh & Co. Kgaa Composition therapeutique avec neurotoxine botulique
WO2009114748A1 (fr) 2008-03-14 2009-09-17 Allergan, Inc. Essais d’activité de sérotype a de toxine du botulisme à base immunitaire
US8398998B2 (en) 1999-06-07 2013-03-19 Merz Pharma Gmbh & Co. Kgaa Therapeutic composition comprising a botulinum neurotoxin
WO2013049508A1 (fr) 2011-09-29 2013-04-04 WHITEMARSH, Regina Clare Meyer Compositions et procédés permettant d'effectuer des tests de toxigénicité
WO2014207109A1 (fr) 2013-06-28 2014-12-31 Merz Pharma Gmbh & Co. Kgaa Moyens et procédés pour déterminer l'activité biologique de polypeptides de neurotoxine dans des cellules
WO2018222652A1 (fr) 2017-05-31 2018-12-06 Allergan, Inc. Neurotoxine botulique pour le traitement de troubles associés à l'hyperactivité des mélanocytes et/ou à la mélanine en excès

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8398998B2 (en) 1999-06-07 2013-03-19 Merz Pharma Gmbh & Co. Kgaa Therapeutic composition comprising a botulinum neurotoxin
WO2005007185A2 (fr) 2003-07-22 2005-01-27 Biotecon Therapeutics Gmbh Formulation destinee a un agent pharmaceutique proteique ne contenant pas d'albumine serique humaine ajoutee (hsa)
WO2006020208A2 (fr) 2004-07-26 2006-02-23 Merz Pharma Gmbh & Co. Kgaa Composition therapeutique avec neurotoxine botulique
WO2009114748A1 (fr) 2008-03-14 2009-09-17 Allergan, Inc. Essais d’activité de sérotype a de toxine du botulisme à base immunitaire
WO2013049508A1 (fr) 2011-09-29 2013-04-04 WHITEMARSH, Regina Clare Meyer Compositions et procédés permettant d'effectuer des tests de toxigénicité
WO2014207109A1 (fr) 2013-06-28 2014-12-31 Merz Pharma Gmbh & Co. Kgaa Moyens et procédés pour déterminer l'activité biologique de polypeptides de neurotoxine dans des cellules
WO2018222652A1 (fr) 2017-05-31 2018-12-06 Allergan, Inc. Neurotoxine botulique pour le traitement de troubles associés à l'hyperactivité des mélanocytes et/ou à la mélanine en excès

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
"Remington's Pharmaceutical Sciences", MACK PUBLISHING COMPANY
DALLAGNOL SOARES ANDRESSA ET AL: "Use of botulinum toxin type A associated with nonthermal microneedling for treating erythematoustelangiectatic rosacea: A case report", JOURNAL OF CLINICAL IMAGES AND MEDICAL CASE REPORTS, vol. 3, no. 7, 18 July 2022 (2022-07-18), XP093110662, ISSN: 2766-7820, DOI: 10.52768/2766-7820/1955 *
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., vol. 128, 1994, pages 69 - 77, XP024880303, DOI: 10.1006/taap.1994.1181
SHIM DOO HEE ET AL: "Development of Botulinum Toxin A-Coated Microneedles for Treating Palmar Hyperhidrosis", vol. 16, no. 12, 2 December 2019 (2019-12-02), US, pages 4913 - 4919, XP093085894, ISSN: 1543-8384, Retrieved from the Internet <URL:http://pubs.acs.org/doi/pdf/10.1021/acs.molpharmaceut.9b00794> DOI: 10.1021/acs.molpharmaceut.9b00794 *
SUKSANTILAP SONGSAK ET AL: "The Efficacy of Intradermal Injection of Botulinum Toxin A In The Treatment of Dermal and Mixed Type, Melasma", HE 4TH NATIONAL ACADEMIC CONFERENCE AND RESEARCH PRESENTATION GRADUATE SCHOOL CONFERENCE 2022 IHAPPINESS: SUSTAINABLE HAPPINESS AND GOOD QUALITY OF LIFE IN THE DIGITAL AGEAT: SUAN SUNANDHA RAJABHAT UNIVERSITY. 31 AUGUST 2022, 1 January 2022 (2022-01-01), XP093112625 *
VACHIRAMON VASANOP ET AL: "Incobotulinum Toxin Type A for Treatment of Ultraviolet-B-Induced Hyperpigmentation: A Prospective, Randomized, Controlled Trial", TOXINS, vol. 14, no. 6, 17 June 2022 (2022-06-17), CH, pages 417, XP093112626, ISSN: 2072-6651, DOI: 10.3390/toxins14060417 *

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