Methods of reducing collagenase mediated bruises in subjects with cellulite

Abstract

Disclosed herein are methods of reducing collagenase mediated bruises in a subject having cellulite, methods of treating cellulite in a subject, and formulations containing collagenase.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 378,424, filed on October 5, 2022, and priority to U.S. Provisional Application No. 63 / 340,295, filed on May 10, 2022, the entire disclosures of which are incorporated herein by reference in their entirety. Technical Field

[0003] Disclosed herein are methods of reducing collagenase-mediated bruising in a subject having cellulite, methods of treating cellulite in a subject, and formulations containing collagenase. Background Art

[0004] Collagenase refers to a protease that hydrolyzes collagen in its native triple-helical conformation under physiological conditions. (CCH-aaes) is a combination of AUX-I and AUX-II (also known as collagenase I and collagenase II) that has been approved for the treatment of moderate to severe cellulite on the buttocks in adult women. It effectively dissolves the fibrous septa located under the dermis at the injection site, which is the root cause of skin pits in women with cellulite. It has been shown to have an acceptable safety profile and immunogenicity, with injection site bruising being the most common adverse reaction. The associated post-injection bruises typically resolved within 21 days, but were distressing to participants due to potential skin discoloration and associated risk of swelling and pain until the next treatment. Summary of the invention

[0005] Disclosed herein are methods of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising subcutaneously administering a total dose of about 0.42 mg of collagenase to the treatment area of ​​the subject per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0006] Disclosed herein is a method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0007] Disclosed herein is a method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising subcutaneously administering a total dose of about 0.84 mg of collagenase to a depth of about 0.25 inches per treatment session to a treatment area of ​​the subject, thereby reducing collagenase-mediated bruising in the subject.

[0008] Disclosed herein is a method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising subcutaneously administering to a treatment area of ​​the subject a total dose of approximately 0.84 mg of collagenase having a concentration of approximately 0.09 mg / ml per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0009] Disclosed herein are methods of reducing collagenase-mediated bruising in a subject having cellulite, the methods comprising subcutaneously administering to a treatment area of ​​the subject a composition comprising collagenase, lidocaine, and epinephrine, wherein a total dose of approximately 0.42 mg of collagenase is administered per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0010] Disclosed herein is a method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of approximately 0.21 mg of collagenase having a concentration of approximately 0.12 mg / ml per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0011] Disclosed herein is a method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising orally administering tranexamic acid (TXA) to the subject and subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of ​​the subject during each treatment period, thereby reducing collagenase-mediated bruising in the subject.

[0012] Disclosed herein is a method for reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising subcutaneously administering a total dose of about 0.42 mg of collagenase to the treatment area of ​​the subject during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0013] Disclosed herein are methods for reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0014] Disclosed herein is a method for reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising subcutaneously administering a total dose of about 0.84 mg of collagenase to a depth of about 0.25 inches per treatment session to the treatment area of ​​the subject, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0015] Disclosed herein are methods for reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.09 mg / ml during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0016] Disclosed herein are methods of reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to a treatment area of ​​the subject, wherein a total dose of approximately 0.42 mg of collagenase is administered during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0017] Disclosed herein are methods for reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.21 mg of collagenase having a concentration of about 0.12 mg / ml during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0018] Disclosed herein are methods of reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in a subject, the method comprising orally administering tranexamic acid to the subject and subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of ​​the subject during each treatment period, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated cellulite treatment in the subject.

[0019] Also provided herein is a method of treating cellulite in a subject, the method comprising subcutaneously administering a total dose of about 0.42 mg of collagenase to the treatment area of ​​the subject during each treatment session, thereby treating cellulite in the subject.

[0020] Disclosed herein are methods of treating cellulite in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml per treatment session, thereby treating cellulite in the subject.

[0021] Disclosed herein is a method for treating cellulite in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.23 mg / ml at a depth of about 0.25 inches per treatment session, thereby treating cellulite in the subject.

[0022] Disclosed herein are methods of treating cellulite in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.09 mg / ml per treatment session, thereby treating cellulite in the subject.

[0023] Disclosed herein is a method of treating cellulite in a subject, the method comprising subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to a treatment area of ​​the subject during each treatment session, wherein a total dose of about 0.42 mg of collagenase is administered during each treatment session, thereby treating cellulite in the subject.

[0024] Disclosed herein are methods of treating cellulite in a subject, the method comprising subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.21 mg of collagenase having a concentration of about 0.12 mg / ml per treatment session, thereby treating cellulite in the subject.

[0025] Also provided herein are formulations comprising collagenase, lidocaine, and epinephrine.

[0026] BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The Abstract and the following Detailed Description will be further understood when read in conjunction with the accompanying drawings. To illustrate the disclosed methods and formulations, exemplary embodiments of the methods and formulations are shown in the accompanying drawings; however, the methods and formulations are not limited to the specific embodiments disclosed herein. In the drawings:

[0028] Figure 1 An exemplary "triple injection technique" (also known as the "three equal parts" technique) is shown, in which the subcutaneous injection is administered in the form of three 0.1 ml aliquots (total injection volume of 0.3 ml), wherein the first aliquot is administered perpendicular to the skin (A) and the other two aliquots are administered above (B) or below (C) at an angle of 45° relative to the vertical axis. Figure 1 The injection depth in the is 0.5 inches, but a 0.25 inch injection depth can also be used. Each syringe contains 0.9 ml, which allows for 3 injections per syringe and a maximum of 12 injections per treatment area.

[0029] Figure 2 Shown is an exemplary Investigator Assessment of Bruising Severity Assessment Scale (IABSS) used to assess the severity of bruising and / or skin discoloration following subcutaneous administration of collagenase to the treatment area (in this example, the buttocks).

[0030] Figure 3 Shown is an exemplary grid injection pattern in which up to 30 injections may be delivered to the treatment area with each injection spaced approximately 3 cm apart.

[0031] Figure 4A and Figure 4B Shown are queues 1 to 5 ( Figure 4A ) and queue 6( Figure 4B ) in the APHRODITE-1 study.

[0032] Figure 5 Shown is the extent of bruising observed in patients from Cohort 1 at Visit 3 (Days 3-5 after treatment). Left buttock (experimental): received 0.42 mg of CCH-aaes at a concentration of 0.23 mg / mL using a three-injection technique. Right buttock (control): received 0.84 mg of CCH-aaes at a concentration of 0.23 mg / mL using a three-injection technique. DETAILED DESCRIPTION

[0033] The disclosed methods and formulations may be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure. It should be understood that the disclosed methods and formulations are not limited to the specific methods and formulations described and / or shown herein, and that the terminology used herein is for the purpose of illustrating specific embodiments only and is not intended to limit the claimed methods and formulations.

[0034] Unless expressly stated otherwise, any description of a possible mechanism or mode of action or reason for improvement is meant to be illustrative only, and the disclosed methods and formulations are not limited by the correctness or incorrectness of any such suggested mechanism or mode of action or reason for improvement.

[0035] In the case where a numerical range is quoted or established in this article, the range includes the endpoints of the value and all single integers and decimals in the range, and also includes each of the narrower ranges formed by all various possible combinations of these endpoints and internal integers and decimals in the range, to form a subgroup of the larger value group in the range, the extent of which is the same as the extent to which each narrower range is explicitly described. When the numerical range described herein is greater than the specified value, the range is still limited, and is limited at its upper end by a value that is operable in the context of the present invention as described herein. In the case where the numerical range described herein is less than the specified value, the lower limit of the range is limited by a non-zero value. When limiting the range, the scope of the method and preparation is not intended to be limited to the specific value described. All ranges are inclusive and combinable.

[0036] It should be understood that, for clarity, certain features of the disclosed methods and formulations described in the context of separate embodiments herein may also be provided in combination with a single embodiment. Conversely, for brevity, various features of the disclosed methods and formulations described in the context of a single embodiment may also be provided individually or in any sub-combination.

[0037] As used herein, the singular form "a," "an," and "the" include plural forms.

[0038] Various terms related to various aspects of the specification are used throughout the specification and claims. Unless otherwise indicated, such terms should be given their usual meanings in the art. Other explicitly defined terms should be interpreted in a manner consistent with the definitions provided herein.

[0039] When used to refer to a numerical range, a cutoff value, or a specific value, the term "about" is used to indicate that the referenced value may differ from the listed value by up to 15%. Therefore, the term "about" is used to include a specified value that differs by ±15% or less, ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less. When a value is expressed as an approximation, by using the antecedent "about", it is understood that the specific value forms another embodiment. Unless the context clearly states otherwise, a reference to a specific numerical value includes at least the specific value.

[0040] The term “comprising” is intended to include, but not necessarily be limited to, examples encompassed by the terms “consisting essentially of” and “consisting of; similarly, the term “consisting essentially of” is intended to include, but not necessarily be limited to, examples encompassed by the term “consisting of.”

[0041] The term "subject" as used herein is intended to mean any animal, particularly a mammal. Thus, these methods are applicable to both human and non-human animals, although most preferably to humans. "Subject" and "patient" are used interchangeably herein.

[0042] As used herein, "administering" and similar terms refer to a process by which the collagenase or a composition / pharmaceutical formulation comprising collagenase is injected into a subject so that target cells, tissues or parts of the subject's body are contacted with the collagenase.

[0043] As used herein, "collagenase-mediated bruising" refers to at least one of bruising, hematoma, hemorrhage, petechiae, and discoloration that occurs at and / or around the injection site following administration of collagenase.

[0044] "Treat," "treatment" and similar terms include reducing and / or eliminating cellulite, reducing and / or eliminating the underlying causes of cellulite, reducing and / or eliminating the potential for cellulite, and inducing and / or improving the overall cosmetic qualities of the skin.

[0045] As used herein, "reduced bruising" and similar phrases refer to an improvement of at least 1 level on the Investigator Assessment of Bruising Severity Scale (IABSS).

[0046] The term "treatment area" refers to an area on a subject having cellulite and to which a specified total dose of collagenase is administered. Exemplary treatment areas include, for example, the left buttocks, the right buttocks, or both buttocks.

[0047] The term "treatment session" is synonymous with "treatment visit" and includes a single visit to a physician's office where collagenase or a formulation / composition comprising collagenase is administered, or if the subject self-administers collagenase or a formulation / composition comprising collagenase, a single self-administration period. A treatment session or a treatment visit may involve treating a single treatment area or treating multiple treatment areas. For example, in the buttocks example, a treatment visit may involve administering a specified total dose of collagenase to both the left buttocks and the right buttocks. Alternatively, a treatment session or a treatment visit may involve administering a specified total dose of collagenase to only the left buttocks.

[0048] The term "total dose" refers to the total amount of collagenase applied to the treatment area in a given treatment session. For example, in the embodiments disclosed herein, the total dose per treatment session refers to the total dose of collagenase applied to a single buttock in each treatment session.

[0049] Methods of reducing collagenase-mediated bruising in subjects with cellulite

[0050] Disclosed herein is a method for reducing collagenase-mediated bruising in a subject having cellulite. The method may include administering a total dose of about 0.42 mg of collagenase subcutaneously to the treatment area of ​​the subject per treatment session, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase may be administered using a three-injection technique (as described herein). In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Thus, the method may include administering a total dose of about 0.42 mg of collagenase subcutaneously to the treatment area of ​​the subject per treatment session at a depth of about 0.5 inches using a three-injection technique, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase may have a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment, wherein the treatment sessions are separated by about 3 weeks. In some embodiments, the subject may receive more than one treatment, wherein the treatment sessions are separated by about 42 days. According to U.S. Prescribing Information (www_accessdata.fda.gov / drugsatfda_docs / label / 2020 / 761146s0001b1.pdf; see also Example 2 herein) for administration In some embodiments, bruising can be reduced compared to levels of bruising associated with subcutaneous administration of a collagenase totaling about 0.84 mg at a depth of 0.5 inches per treatment session to the subject's treatment area using a three-injection technique. In some embodiments, bruising can be reduced compared to levels of bruising associated with subcutaneous administration of a collagenase totaling about 0.84 mg at a depth of 0.5 inches per treatment session to the subject's treatment area using a three-injection technique, wherein the collagenase has a concentration of about 0.23 mg / ml.

[0051] A method for reducing collagenase-mediated bruising in a subject having cellulite may comprise administering a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at each treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Thus, a method for reducing collagenase-mediated bruising in a subject having cellulite may comprise administering a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.5 inches using a three-injection technique at each treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the subject may receive more than one treatment, wherein the treatment sessions are separated by about 3 weeks. U.S. Prescribing Information for Administration In some embodiments, the collagenase has a concentration of about 0.23 mg / ml and bruising is reduced compared to levels of bruising associated with bruising in patients with sham-treated subjects. In some embodiments, the collagenase has a concentration of about 0.23 mg / ml and bruising is reduced compared to a total dose of about 0.84 mg of collagenase administered subcutaneously to the treatment area of ​​the subject at about 0.5 inches per treatment session using a three-injection technique.

[0052] A method for reducing collagenase-mediated bruising in a subject having cellulite may comprise administering a total dose of about 0.84 mg of collagenase subcutaneously to a treatment area of ​​the subject at a depth of about 0.25 inches per treatment session, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. Thus, a method for reducing collagenase-mediated bruising in a subject having cellulite may comprise administering a total dose of about 0.84 mg of collagenase subcutaneously to a treatment area of ​​the subject at a depth of about 0.25 inches per treatment session, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase may have a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment, wherein the treatment sessions are separated by about 3 weeks. U.S. Prescribing Information for Administration In some embodiments, bruising can be reduced compared to the level of bruising associated with subcutaneous administration of a collagenase totaling about 0.84 mg at a depth of about 0.5 inches per treatment session to the subject's treatment area using a three-injection technique. In some embodiments, the collagenase has a concentration of about 0.23 mg / ml, and the degree of bruising can be reduced compared to the level of bruising associated with subcutaneous administration of a collagenase totaling about 0.84 mg at a depth of about 0.5 inches per treatment session to the subject's treatment area using a three-injection technique.

[0053] Figure 1 An exemplary three-injection technique is shown in . As shown, the subcutaneous injection can be divided into three 0.1 ml aliquots (total injection volume of 0.3 ml), where 1 aliquot is applied perpendicular to the skin (A) and the other 2 aliquots are applied above (B) or below (C) at 45 degrees from the vertical axis. Figure 1 The injection depth in the is 0.5 inches, but other injection depths, such as 0.25 inches, can also be used. Each syringe can hold 0.9 ml, and each syringe can be used for 3 injections, up to 12 injections per treatment area.

[0054] The volume of each aliquot of injection depends in part on the severity of the fat mass treated and the size of the treatment area. The total volume to be injected can be evenly distributed in each aliquot. For example, if 0.3 ml of a pharmaceutical preparation is to be injected, each of the three aliquots can contain 0.1 ml. The suitable volume of each aliquot includes, for example, about 0.05 ml, about 0.075 ml, about 0.1 ml, about 0.2 ml, about 0.3 ml, about 0.4 ml, about 0.5 ml, about 0.6 ml, about 0.7 ml, about 0.8 ml, about 0.9 ml or about 1.0 ml. In some embodiments, each aliquot has a volume of about 0.05 ml to about 0.5 ml.

[0055] A method of reducing collagenase-mediated bruises in a subject with cellulite may include administering a total dose of about 0.84 mg of collagenase subcutaneously to the subject's treatment area at each treatment stage, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing collagenase-mediated bruises in the subject. In some embodiments, the collagenase may be administered using a single aliquot injection. In some embodiments, the collagenase may be administered at a depth of about 0.25 inches. Therefore, a method of reducing collagenase-mediated bruises in a subject with cellulite may include administering a total dose of about 0.84 mg of collagenase subcutaneously to the subject's treatment area at each treatment stage, the collagenase having a concentration of about 0.09 mg / ml, and a single aliquot injection at a depth of about 0.25 inches, thereby reducing collagenase-mediated bruises in the subject. In some embodiments, administration includes injecting up to 30 collagenases into the treatment area in a single aliquot at each treatment stage. In some embodiments, administration includes injecting more than 30 collagenases into the treatment area in a single aliquot at each treatment stage. In some embodiments, a subject may receive more than one treatment, with treatment sessions separated by about 3 weeks. U.S. Prescribing Information for Administration In some embodiments, bruising can be reduced compared to the level of bruising associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase to the subject's treatment area per treatment session, wherein the collagenase has a concentration of about 0.23 mg / ml and is administered in a single aliquot injection to a depth of about 0.25 inches. In some embodiments, the administration comprises up to 12 single aliquot injections of collagenase to the treatment area per treatment session. In some embodiments, the administration comprises more than 12 single aliquot injections of collagenase to the treatment area per treatment session.

[0056] The number of injections administered via a single aliquot injection in a single treatment session depends in part on the severity of the cellulite and the size of the treatment area. In a single treatment session, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more injections may be administered to the treatment area.

[0057] When multiple injections are administered in a single treatment phase, the injections may be spaced evenly or approximately evenly. The intervals between injections depend in part on the amount injected and the size of the treatment area. The injection intervals may, for example, be about 0.25 cm, about 0.5 cm, about 0.75 cm, about 1.0 cm, about 1.25 cm, about 1.5 cm, about 1.75 cm, about 2.0 cm, about 2.25 cm, about 2.5 cm, about 2.75 cm, about 3.0 cm, about 3.25 cm, about 3.5 cm, about 3.75 cm, about 4.0 cm, or more than 4.0 cm. In some embodiments, the injection interval is about 2 cm to about 3 cm.

[0058] When multiple injections are administered in a single treatment session, the injections can be administered at random sites within the treatment area, as one or more patterns within the treatment area, or at random sites within the treatment area and as one or more dot patterns. In some embodiments, the injections are spaced in a grid pattern. Figure 3 An exemplary grid pattern is shown in . As used herein, the grid pattern refers to a repeatable injection pattern with injections evenly spaced. The grid pattern can be hexagonal, circular, square, rectangular, triangular, etc. In some embodiments, the injection intervals in the grid pattern are about 2 cm to about 3 cm. When collagenase or a pharmaceutical preparation / composition comprising collagenase is injected at random sites, in one or more patterns, or in one or more patterns at random locations, the injections can include injecting a single aliquot or multiple aliquots of the pharmaceutical preparation into each location within the treatment area.

[0059] Suitable volumes for each injection within the grid pattern include, for example, about 0.1 ml, about 0.2 ml, about 0.3 ml, about 0.4 ml, about 0.5 ml, about 0.6 ml, about 0.7 ml, about 0.8 ml, about 0.9 ml, or about 1.0 ml. In some embodiments, the volume of each injection is about 0.1 ml to 0.3 ml.

[0060] The collagenase in any method of reducing collagenase-mediated bruising disclosed herein can be a component of a composition / pharmaceutical formulation. In some embodiments, the collagenase can be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation may also comprise HCl to adjust the pH as needed. The pH value of the composition / pharmaceutical formulation may be about 8.0.

[0061] The method of reducing collagenase-mediated bruises in a subject with cellulite may include subcutaneously applying a composition comprising collagenase, lidocaine and epinephrine to the treatment area of ​​the subject, wherein each treatment stage uses a total dose of about 0.42 milligrams of collagenase, thereby reducing collagenase-mediated bruises in the subject. In some embodiments, the collagenase may be applied using a three-injection technique. In some embodiments, the collagenase may be applied at a depth of about 0.5 inches. Therefore, the method of reducing collagenase-mediated bruises in a subject with cellulite may include subcutaneously applying a composition comprising collagenase, lidocaine and epinephrine to the treatment area of ​​the subject, wherein each treatment stage uses a three-injection technique at a depth of about 0.5 inches to apply a total dose of about 0.42 milligrams of collagenase, thereby reducing collagenase-mediated bruises in the subject. The collagenase may have a concentration of about 0.23 mg / ml. The concentration of lidocaine may be about 5 mg / ml, about 10 mg / ml, about 15 mg / ml, or about 20 mg / ml. The concentration of epinephrine may be about 5 μg / ml or about 10 μg / ml. In some embodiments, the composition may include about 0.23 mg / ml collagenase, 2% lidocaine (20 mg / ml), and 1:200,000 epinephrine (5 μg / ml). In some embodiments, the composition may also include mannitol, sucrose, and tromethamine. Thus, the method may include subcutaneously administering to the treatment area of ​​the subject a composition including collagenase, lidocaine, epinephrine, mannitol, sucrose, and tromethamine, wherein a total dose of about 0.42 mg of collagenase is administered per treatment session, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the subject may receive more than one treatment session, wherein the treatment sessions are separated by about 3 weeks. U.S. Prescribing Information for Administration In some embodiments, bruising can be reduced compared to the level of bruising associated with subcutaneously administering a composition comprising collagenase to the treatment area of ​​the subject, wherein a total dose of about 0.84 mg of collagenase is administered at each treatment session using a three-injection technique at a depth of about 0.5 inches. In some embodiments, bruising can be reduced compared to the level of bruising associated with subcutaneously administering a composition comprising about 0.23 mg / ml of collagenase to the treatment area of ​​the subject, wherein the composition is administered at a depth of about 0.5 inches using a three-injection technique, and the total dose of collagenase administered at each treatment session is about 0.84 mg. In some embodiments, bruising is reduced compared to the level of bruising associated with subcutaneously administering a composition comprising collagenase, mannitol, sucrose, and tromethamine to the treatment area of ​​the subject, wherein the composition is administered at a depth of about 0.5 inches using a three-injection technique, and the total dose of collagenase administered at each treatment session is about 0.84 mg.

[0062] A method for reducing collagenase-mediated bruising in a subject having cellulite may include administering a total dose of about 0.21 mg of collagenase subcutaneously to the treatment area of ​​the subject during each treatment session, the collagenase having a concentration of about 0.12 mg / ml, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Thus, a method for reducing collagenase-mediated bruising in a subject having cellulite may include administering a total dose of about 0.21 mg of collagenase subcutaneously to the treatment area of ​​the subject during each treatment session, the collagenase having a concentration of about 0.12 mg / ml, at a depth of about 0.5 inches, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the subject may receive more than one treatment session, wherein the treatment sessions are separated by about 42 days. U.S. Prescribing Information for Administration In some embodiments, bruising can be reduced compared to the level of bruising associated with subcutaneous administration of a total dose of about 0.42 mg of collagenase at a depth of about 0.5 inches to the subject's treatment area using a three-injection technique, wherein the collagenase concentration is about 0.23 mg / ml and the subject receives more than one treatment session separated by about 42 days.

[0063] The method of reducing collagenase-mediated bruises in a subject with cellulite may include orally administering tranexamic acid to a subject, and applying a total dose of about 0.84 mg of collagenase subcutaneously to the subject's treatment area at each treatment stage, thereby reducing collagenase-mediated bruises in the subject. In some embodiments, the concentration of tranexamic acid is about 500 mg to about 1500 mg. In some embodiments, tranexamic acid has a concentration of about 1300 mg. Therefore, the method of reducing collagenase-mediated bruises in a subject with cellulite may include orally administering about 1300 mg of tranexamic acid to a subject, and applying a total dose of about 0.84 mg of collagenase subcutaneously to the subject's treatment area at each treatment stage, thereby reducing collagenase-mediated bruises in the subject. In some embodiments, collagenase can be administered using a three-injection technique. In some embodiments, collagenase can be administered at a depth of about 0.5 inches. The method of reducing collagenase-mediated bruises in a subject with cellulite can include orally administering about 1300 mg of tranexamic acid to the subject, and using a three-injection technique to subcutaneously administer a total dose of about 0.84 mg of collagenase to the subject's treatment area at a depth of about 0.5 inches, thereby reducing collagenase-mediated bruises in the subject. In some embodiments, the collagenase can have a concentration of about 0.23 mg / ml. In some embodiments, the subject can receive more than one treatment phase, wherein the treatment phase is spaced about 3 weeks apart. In some embodiments, in the first treatment phase, tranexamic acid is taken orally three times a day for five days. In some aspects, in the first treatment phase, the subject orally administers tranexamic acid three times a day on the day before the application of collagenase, the day of the application of collagenase, and the 2nd, 3rd, and 4th days after the application of collagenase. In some embodiments, in the second treatment phase, tranexamic acid is taken orally three times a day for five days. In certain aspects, in the second treatment period, the subject is orally administered tranexamic acid three times daily on the day before, on the day of, and on the 2nd, 3rd, and 4th day after the administration of collagenase. U.S. Prescribing Information for Administration In some embodiments, bruising can be reduced compared to levels of bruising associated with subcutaneous administration of collagenase using a three-injection technique at a depth of 0.5 inches to the treatment area of ​​the subject, with a total dose of about 0.84 mg of collagenase being administered subcutaneously.

[0064] The level of bruising can be assessed using the Investigator Assessment of Bruising Severity Scale (IABSS). Figure 2 An exemplary IABSS is shown in FIG. Figure 2As shown, the exemplary IABSS includes 5 images showing bruises in the treatment area (left buttocks and right buttocks) of one or more exemplary subjects, the treatment area in each of the 5 images of the exemplary subject corresponding to the location of the treatment area of ​​the subject to whom collagenase was administered, wherein each of the 5 images has a different bruise severity rating, including a numerical value and a description. The exemplary IABSS includes the following severity ratings:

[0065] 0-none or almost none: no bruising or almost no bruising was observed;

[0066] 1-Mild: Mild bruising, pink to light red covering about 25% of the buttocks;

[0067] 2-Moderate: Moderate bruising, dark red to purple covering about 50% of the buttocks;

[0068] 3- Severe: Severe bruising, dark purple covering approximately 75% of the buttocks; and

[0069] 4 - Very severe: Severe bruising, dark purple to deepest purple covering approximately 100% of the buttocks.

[0070] A reduction in the severity of bruising can include an improvement (i.e., a reduction) in at least one point of the IABSS. Suitable reductions include, for example, a severity rating of 4 to 3, a severity rating of 3 to 2, a severity rating of 2 to 1, a severity rating of 1 to 0, a severity rating of 4 to 2, a severity rating of 4 to 1, a severity rating of 4 to 0, etc.

[0071] Methods of reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject

[0072] Disclosed herein is a method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject. The method may include administering a total dose of about 0.42 mg of collagenase subcutaneously to the treatment area of ​​the subject during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Thus, the method may include administering a total dose of about 0.42 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.5 inches during each treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase has a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment session, wherein the treatment sessions are separated by about 3 weeks. In some embodiments, the subject may receive more than one treatment session, wherein the treatment sessions are separated by about 42 days. According to U.S. Prescribing Information for Administration In some embodiments, bruising and / or skin discoloration can be reduced compared to the levels of bruising and / or skin discoloration associated with subcutaneous administration of a collagenase to the treatment area of ​​the subject using a three-injection technique at a depth of 0.5 inches per treatment session, and bruising and / or skin discoloration can be reduced. In some embodiments, the collagenase has a concentration of about 0.23 mg / ml, and bruising and / or skin discoloration can be reduced compared to the levels of bruising and / or skin discoloration associated with subcutaneous administration of a collagenase to the treatment area of ​​the subject using a three-injection technique at a depth of 0.5 inches per treatment session, and bruising and / or skin discoloration can be reduced.

[0073] The method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include subcutaneously applying a total dose of about 0.84 mg of collagenase to the subject's treatment area, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase may be applied using a three-injection technique. In some embodiments, the collagenase may be applied at a depth of about 0.5 inches. Therefore, the method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include subcutaneously applying a total dose of about 0.84 mg of collagenase to the subject's treatment area at a depth of about 0.5 inches, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the subject may receive more than one treatment phase, wherein the treatment phases are spaced about 3 weeks apart. According to U.S. Prescribing Information for Administration In some embodiments, the collagenase has a concentration of about 0.23 mg / ml, bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase per treatment session to the subject using a three-injection technique at a depth of about 0.5 inches.

[0074] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may comprise administering a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.25 inches using a three-injection technique per treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. Thus, a method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may comprise administering a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.25 inches using a three-injection technique per treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase has a concentration of about 0.23 mg / mL. In some embodiments, the subject may receive more than one treatment session, wherein the treatment sessions are separated by about 3 weeks. U.S. Prescribing Information for Administration In some embodiments, the collagenase has a concentration of approximately 0.23 mg / ml, and the degree of bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a collagenase totaling approximately 0.84 mg per treatment session to the subject's treatment area using a three-injection technique at a depth of approximately 0.5 inches. In some embodiments, the collagenase has a concentration of approximately 0.23 mg / ml, and the degree of bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a collagenase totaling approximately 0.84 mg per treatment session to the subject's treatment area using a three-injection technique at a depth of approximately 0.5 inches.

[0075] The volume of each aliquot of injection depends in part on the severity of the fat mass treated and the size of the treatment area. The total volume to be injected can be evenly distributed in each aliquot. For example, if 0.3 ml of a pharmaceutical preparation is to be injected, each of the three aliquots can contain 0.1 ml. The suitable volume of each aliquot includes, for example, about 0.05 ml, about 0.075 ml, about 0.1 ml, about 0.2 ml, about 0.3 ml, about 0.4 ml, about 0.5 ml, about 0.6 ml, about 0.7 ml, about 0.8 ml, about 0.9 ml or about 1.0 ml. In some embodiments, each aliquot has a volume of about 0.05 ml to about 0.5 ml.

[0076] The method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject per treatment phase, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase may be applied using a single aliquot injection technique. In some embodiments, the collagenase may be applied at a depth of about 0.25 inches. Therefore, the method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject per treatment phase using a single aliquot injection technique at a depth of about 0.25 inches, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the administration comprises up to 30 aliquot injections of collagenase to the treatment area per treatment session. In some embodiments, the administration comprises more than 30 aliquot injections of collagenase to the treatment area per treatment session. In some embodiments, the subject may receive more than a single treatment session, wherein the treatment sessions are spaced approximately 3 weeks apart. U.S. Prescribing Information for Administration In some embodiments, the collagenase has a concentration of approximately 0.23 mg / ml, and bruises and / or skin discoloration can be reduced compared to the level of bruises and / or skin discoloration associated with subcutaneous administration of a total dose of approximately 0.84 mg of collagenase to the treatment area of ​​the subject at a depth of approximately 0.25 inches per treatment session using a single aliquot injection technique. In some embodiments, the administration includes up to 12 single aliquot injections of collagenase to the treatment area per treatment session. In some embodiments, the administration includes more than 12 single aliquot injections of collagenase to the treatment area per treatment session.

[0077] The number of injections administered via a single aliquot injection in a single treatment session depends in part on the severity of the cellulite and the size of the treatment area. In a single treatment session, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more injections may be administered to the treatment area.

[0078] When multiple injections are administered in a single treatment course, the injections can be spaced evenly or approximately evenly. The intervals between injections depend in part on the volume injected and the size of the treatment area. The injection intervals can be, for example, about 0.25 cm, about 0.5 cm, about 0.75 cm, about 1.0 cm, about 1.25 cm, about 1.5 cm, about 1.75 cm, about 2.0 cm, about 2.25 cm, about 2.5 cm, about 2.75 cm, about 3.0 cm, about 3.25 cm, about 3.5 cm, about 3.75 cm, about 4.0 cm, or more than 4.0 cm. In some embodiments, the injection interval is about 2 cm to about 3 cm.

[0079] When multiple injections are administered in a single treatment session, the injections can be administered at random sites within the treatment area, as one or more patterns within the treatment area, or at random sites and as one or more dot patterns within the treatment area. In some embodiments, the injections are spaced apart in a grid pattern. In some embodiments, the injections in the grid pattern are spaced apart by about 2 centimeters to about 3 centimeters. When the collagenase or a pharmaceutical formulation / composition comprising the collagenase is injected at random sites, as one or more patterns, or at random sites and as one or more patterns, the injections can include injecting a single aliquot or multiple aliquots of the pharmaceutical formulation into each site within the treatment area.

[0080] Suitable volumes for each injection within the grid pattern include, for example, about 0.1 ml, about 0.2 ml, about 0.3 ml, about 0.4 ml, about 0.5 ml, about 0.6 ml, about 0.7 ml, about 0.8 ml, about 0.9 ml, or about 1.0 ml. In some embodiments, the volume of each injection is about 0.1 ml to 0.3 ml.

[0081] The collagenase in any of the methods for reducing bruises and / or skin discoloration disclosed herein can be a component of a composition / pharmaceutical formulation. In some embodiments, the collagenase can be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation may also comprise HCl to adjust the pH as needed. The pH of the composition / pharmaceutical formulation may be about 8.0.

[0082] A method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to a treatment area of ​​the subject, wherein a total dose of about 0.42 milligrams of collagenase is administered at each treatment stage, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Therefore, a method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to a treatment area of ​​the subject, wherein a total dose of about 0.42 milligrams of collagenase is administered at each treatment stage using a three-injection technique at a depth of about 0.5 inches, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. The concentration of collagenase is about 0.23 mg / ml. The concentration of lidocaine can be about 5 mg / ml, about 10 mg / ml, about 15 mg / ml or about 20 mg / ml. The concentration of epinephrine can be about 5 μg / ml or about 10 μg / ml. In some embodiments, the composition may include about 0.23 mg / ml collagenase, 2% lidocaine (20 mg / ml) and 1:200,000 epinephrine (5 μg / ml). In some embodiments, the composition also includes mannitol, sucrose and tromethamine. Therefore, the method can include subcutaneously administering a composition comprising collagenase, lidocaine, epinephrine, mannitol, sucrose and tromethamine to the treatment area of ​​the subject, wherein a total dose of about 0.42 mg of collagenase is administered at a depth of about 0.5 inches using a three-injection technique in each treatment phase, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated cellulite treatment of the subject. In some embodiments, a subject may receive more than one treatment period, wherein the treatment periods are separated by about 3 weeks. U.S. Prescribing Information for Administration In some embodiments, bruises and / or skin discoloration can be reduced compared to the level of bruises and / or skin discoloration associated with subcutaneous administration of a composition comprising collagenase to the treatment area of ​​the subject, wherein a total dose of about 0.84 mg of collagenase is administered at a depth of about 0.5 inches using a three-injection technique per treatment session. In some embodiments, bruises and / or skin discoloration can be reduced compared to the level of bruises and / or skin discoloration associated with subcutaneous administration of a composition comprising about 0.23 mg / ml of collagenase to the treatment area of ​​the subject, wherein a total dose of about 0.84 mg of collagenase is administered at a depth of about 0.5 inches using a three-injection technique per treatment session. In some embodiments, bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a composition comprising collagenase, mannitol, sucrose, and tromethamine to the treatment area of ​​a subject, wherein a total dose of about 0.84 mg of collagenase is administered per treatment session using a three-injection technique at a depth of about 0.5 inches.

[0083] A method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include administering a total dose of about 0.21 mg of collagenase subcutaneously to the subject's treatment area at a concentration of 0.12 mg / ml per treatment phase, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject. In some embodiments, the collagenase may be administered using a three-injection technique. In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Therefore, a method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include administering a total dose of about 0.21 mg of collagenase subcutaneously to the subject's treatment area at a depth of about 0.5 inches using a three-injection technique at a depth of about 0.5 inches, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject. In some embodiments, a subject may receive more than one treatment period, wherein the treatment periods are separated by approximately 42 days. U.S. Prescribing Information for Administration In some embodiments, bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with subcutaneously administering a total dose of 0.42 mg of collagenase to the subject's treatment area in each treatment session using a three-injection technique at a depth of about 0.5 inches. In some embodiments, bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with subcutaneously administering a total dose of 0.42 mg of collagenase to the subject's treatment area in each treatment session using a three-injection technique at a depth of about 0.5 inches, wherein the collagenase concentration is about 0.23 mg / ml, wherein the subject receives more than one treatment session, and the treatment sessions are separated by approximately 42 days.

[0084] The method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include orally administering tranexamic acid to the subject and subcutaneously administering a total dose of about 0.84 mg of collagenase to the subject's treatment area, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the concentration of tranexamic acid is about 500 mg to about 1500 mg. In some embodiments, the concentration of tranexamic acid is about 1300 mg. Therefore, the method of reducing bruises and / or skin discoloration associated with collagenase-mediated cellulite treatment of a subject may include orally administering about 1300 mg of tranexamic acid to the subject and subcutaneously administering a total dose of about 0.84 mg of collagenase to the subject's treatment area at each treatment stage, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated cellulite treatment of the subject. In some embodiments, collagenase can be administered using a three-injection technique. In some embodiments, collagenase can be administered at a depth of about 0.5 inches. A method of reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include orally administering about 1300 mg of tranexamic acid to a subject and subcutaneously administering a total dose of about 0.84 mg of collagenase to the subject's treatment area at a depth of about 0.5 inches using a three-injection technique in each treatment phase, thereby reducing bruises and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the concentration of collagenase may be about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment phase, wherein the treatment phases are spaced about 3 weeks apart. In some embodiments, in a first treatment phase, tranexamic acid is orally administered three times a day for five days. In some aspects, in a first treatment phase, tranexamic acid is orally administered to a subject three times a day on the day before collagenase is administered, on the day of collagenase administration, and on the 2nd, 3rd, and 4th day after collagenase administration. In some embodiments, in a second treatment phase, tranexamic acid is orally administered three times a day for five days. In some aspects, during the second treatment period, the subject is orally administered tranexamic acid three times daily on the day before the collagenase is administered, on the day of the collagenase administration, and on the 2nd, 3rd, and 4th day after the collagenase administration. U.S. Prescribing Information for Administration In some embodiments, bruising and / or skin discoloration is reduced compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase to the treatment area of ​​the subject at each treatment session using a three-injection technique at a depth of about 0.5 inches.

[0085] A reduction in the level of bruising and / or skin discoloration can include an improvement (i.e., a decrease) of at least one point on the IABSS. Suitable reductions include, for example, a severity rating of 4 to 3, a severity rating of 3 to 2, a severity rating of 2 to 1, a severity rating of 1 to 0, a severity rating of 4 to 2, a severity rating of 4 to 1, a severity rating of 4 to 0, etc.

[0086] Treatments for cellulite

[0087] Disclosed herein is a method for treating cellulite in a subject. The method for treating cellulite in a subject may include applying a total dose of about 0.42 mg of collagenase subcutaneously to the treatment area of ​​the subject in each treatment phase, thereby treating the cellulite in the subject. In some embodiments, the collagenase may be applied using a three-injection technique. In some embodiments, the collagenase may be applied at a depth of about 0.5 inches. Therefore, the method for treating cellulite in a subject may include applying a total dose of about 0.42 mg of collagenase subcutaneously to the treatment area of ​​the subject in each treatment phase using a three-injection technique at a depth of about 0.5 inches, thereby treating the cellulite in the subject. In some embodiments, the concentration of collagenase is about 0.23 mg / ml. In some embodiments, collagenase may be a component of a composition / pharmaceutical preparation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical preparation may also include HCl to adjust the pH value as needed. The pH value of the composition / pharmaceutical preparation may be about 8.0. In some embodiments, the subject may receive more than one treatment phase, wherein the treatment phases are spaced about 3 weeks apart. In some embodiments, a subject may receive more than one treatment period, wherein the treatment periods are separated by approximately 42 days.

[0088] The method for treating cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at each treatment stage, and the concentration of the collagenase is about 0.05 mg / ml, thereby treating the cellulite in the subject. In some embodiments, the collagenase can be applied using a three-injection technique. In some embodiments, the collagenase can be applied at a depth of about 0.5 inches. Therefore, the method for treating cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.5 inches using a three-injection technique at each treatment stage, and the concentration of the collagenase is about 0.05 mg / ml, thereby treating the cellulite in the subject. In some embodiments, the collagenase can be a component of a composition / pharmaceutical preparation comprising collagenase, mannitol, sucrose and tromethamine. The composition / pharmaceutical preparation may also include HCl to adjust the pH as needed. The pH value of the composition / pharmaceutical preparation may be about 8.0. In some embodiments, the subject may receive more than one treatment stage, wherein the treatment stages are spaced about 3 weeks apart.

[0089] The method for treating cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.25 inches per treatment stage, and the concentration of collagenase is about 0.23 mg / ml, thereby treating the cellulite in the subject. In some embodiments, the collagenase can be applied using a three-injection technique. Therefore, the method for treating cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.25 inches per treatment stage, and the concentration of collagenase is about 0.23 mg / ml, thereby treating the cellulite in the subject. In some embodiments, the collagenase can be a component of a composition / pharmaceutical preparation comprising collagenase, mannitol, sucrose and tromethamine. The composition / pharmaceutical preparation may also include HCl to adjust the pH as needed. The pH value of the composition / pharmaceutical preparation may be about 8.0. In some embodiments, the subject may receive more than one treatment stage, wherein the treatment stages are spaced about 3 weeks apart.

[0090] The method for treating cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at each treatment stage, and the concentration of the collagenase is about 0.09 mg / ml, thereby treating the cellulite in the subject. In some embodiments, the collagenase can be administered using a single aliquot injection. In some embodiments, the collagenase can be administered at a depth of about 0.25 inches. Therefore, the method for treating cellulite in a subject may include applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject at a depth of about 0.25 inches using a single aliquot injection at each treatment stage, and the concentration of the collagenase is about 0.09 mg / ml, thereby treating the cellulite in the subject. In some embodiments, the collagenase can be a component of a composition / pharmaceutical preparation comprising collagenase, mannitol, sucrose and tromethamine. The composition / pharmaceutical preparation may also include HCl to adjust the pH as needed. The pH value of the composition / pharmaceutical preparation may be about 8.0. In some embodiments, the subject may receive more than one treatment stage, wherein the treatment stages are separated by about 3 weeks.

[0091] The number of injections administered by a single aliquot injection in a treatment session depends in part on the severity of the cellulite and the size of the treatment area. In a treatment session, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more injections may be administered to the treatment area. In some embodiments, administration comprises up to 30 single aliquot injections of collagenase to the treatment area in each treatment session.

[0092] When multiple injections are administered in a single treatment phase, the injections can be spaced evenly or approximately evenly. The intervals between injections depend in part on the volume injected and the size of the treatment area. The injection intervals can be, for example, about 0.25 cm, about 0.5 cm, about 0.75 cm, about 1.0 cm, about 1.25 cm, about 1.5 cm, about 1.75 cm, about 2.0 cm, about 2.25 cm, about 2.5 cm, about 2.75 cm, about 3.0 cm, about 3.25 cm, about 3.5 cm, about 3.75 cm, about 4.0 cm, or more than 4.0 cm. In some embodiments, the injection intervals are about 2 cm to about 3 cm.

[0093] When multiple injections are administered in a single treatment session, the injections can be administered at random locations within the treatment area, as one or more patterns within the treatment area, or at random portions within the treatment area and as one or more patterns. In some embodiments, the injections are spaced apart in a grid pattern as described herein.

[0094] Suitable volumes for each injection within the grid pattern include, for example, about 0.1 ml, about 0.2 ml, about 0.3 ml, about 0.4 ml, about 0.5 ml, about 0.6 ml, about 0.7 ml, about 0.8 ml, about 0.9 ml, or about 1.0 ml. In some embodiments, the volume of each injection is about 0.1 ml to 0.3 ml.

[0095] A method for treating cellulite in a subject may include subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to a treatment area of ​​the subject, wherein a total dose of about 0.42 mg of collagenase is administered at each treatment stage, thereby treating cellulite in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. In some embodiments, the collagenase may be administered at a depth of about 0.5 inches. Thus, a method for treating cellulite in a subject may include subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to a treatment area of ​​the subject, wherein a total dose of about 0.42 mg of collagenase is administered at a depth of about 0.5 inches using a three-injection technique at each treatment stage, thereby treating cellulite in the subject. In some embodiments, the composition comprises 0.23 mg / ml of collagenase, 2% lidocaine, and 1:200,000 of epinephrine. In some embodiments, the subject may receive more than one treatment stage, wherein the treatment stages are separated by about 3 weeks. In some embodiments, the composition further comprises mannitol, sucrose, and tromethamine. Thus, a method of treating cellulite in an individual may include subcutaneously administering a composition comprising collagenase, lidocaine, epinephrine, mannitol, sucrose, and tromethamine to a treatment area of ​​the individual, wherein a three-injection technique is used at a depth of approximately 0.5 inches using a total dose of approximately 0.42 mg of collagenase per treatment session, thereby treating cellulite in the subject.

[0096] The method for treating cellulite in a subject may include applying a total dose of about 0.21 mg of collagenase subcutaneously to the treatment area of ​​the subject per treatment phase, and the concentration of collagenase is about 0.12 mg / ml, thereby treating cellulite in the subject. In some embodiments, collagenase can be applied using a three-injection technique. In some embodiments, collagenase can be applied at a depth of about 0.5 inches. Therefore, the method for treating cellulite in a subject may include using a three-injection technique at a depth of about 0.5 inches, and applying a total dose of about 0.21 mg of collagenase subcutaneously to the treatment area of ​​the subject per treatment phase, and the concentration of collagenase is about 0.12 mg / ml, thereby treating cellulite in the subject. In some embodiments, the subject may receive more than one treatment phase, wherein the treatment phases are spaced about 42 days apart. In some embodiments, collagenase may be a component of a composition / pharmaceutical preparation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical preparation may also include HCl to adjust the pH as needed. The pH value of the composition / pharmaceutical preparation may be about 8.0.

[0097] The method for treating cellulite in a subject may include having the subject orally take tranexamic acid, and applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject in each treatment phase, thereby treating the cellulite in the subject. In some embodiments, the concentration of tranexamic acid is about 500 mg to about 1500 mg. In some embodiments, the concentration of tranexamic acid is about 1300 mg. Therefore, the method for treating cellulite in a subject may include having the subject orally take about 1300 mg of tranexamic acid, and applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject in each treatment phase, thereby treating the cellulite in the subject. In some embodiments, the collagenase may be applied using a three-injection technique. In some embodiments, the collagenase may be applied at a depth of about 0.5 inches. The method for treating cellulite in a subject may include having the subject orally take about 1300 mg of tranexamic acid, and applying a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject in each treatment phase at a depth of about 0.5 inches using a three-injection technique, thereby treating the cellulite in the subject. In some embodiments, the concentration of collagenase is about 0.23 mg / ml. In some embodiments, the subject can receive more than one treatment phase, wherein the treatment phase is spaced about 3 weeks apart. In some embodiments, during the first treatment phase, tranexamic acid is orally administered three times a day for 5 days. In some aspects, during the first treatment phase, tranexamic acid is orally administered three times to the subject the day before collagenase is applied, tranexamic acid is orally administered three times to the subject on the day when collagenase is applied, and tranexamic acid is orally administered once to the subject on the 2nd day, 3rd day, and 4th day after collagenase is applied. In some embodiments, during the second treatment phase, tranexamic acid is orally administered three times a day for 5 days. In some aspects, during the second treatment phase, tranexamic acid is orally administered three times to the subject the day before collagenase is applied, tranexamic acid is orally administered three times to the subject on the day when collagenase is applied, and tranexamic acid is orally administered once to the subject on the 2nd day, 3rd day, and 4th day after collagenase is applied.

[0098] Treatment of cellulite may be assessed using a scale or measurement tool selected from the following: Hexsel Cellulite Severity Scale (Hexsel CSS), Hexsel Dimple Depth Rating Scale, Likert Scale, Dimple Analysis, Clinician-Reported Photo-Digital Cellulite Severity Scale (CR-PCSS), Patient-Reported Photo-Digital Cellulite Severity Scale (PR-PCSS), Investigator Global Aesthetic Improvement Scale (I-GAIS), Subject Global Aesthetic Improvement Scale (S-GAIS), Patient-Reported Cellulite Impact Scale (PR-CIS), PR-CIS Simplified, Patient Self-Rating Scale (SSRS), Subject Satisfaction with Cellulite Treatment Questionnaire (SSCT), Clinician Assessment of Cellulite Severity (Photographic or Other Images), Body-Q, and a validated photo-digital or other scale for clinical and / or patient use to assess cellulite severity, improvement, and / or patient satisfaction.

[0099] The Fitzpatrick Skin Scale is a 6-grade scale (I-VI) used to assess skin color and tendency to tan to classify skin type. Skin types range from Grade I: light white skin, blue / hazel eyes, blond / red hair, often tans, never tans, to Grade VI: dark brown or black skin, never tans, only tans. The Investigator (or designee) will determine the Fitzpatrick skin type of all subjects at Screening.

[0100] The Hexsel Cellulite Severity Scale (CSS) is a photographic numeric scale that assesses 5 key morphological features of cellulite: (A) the number of visible dimples, (B) the depth of the dimples, (C) the morphological appearance of changes in the skin surface, (D) the degree of skin looseness, softness, or sagging, and (E) a current classification scale based on the medical literature (Hexsel et al., 2009; Nürnberger and Müüller, 1978). Each of these features is assessed on a 4-point scale, ranging from a low of 0 to a high of 3 (as shown in Table 1).

[0101] Table 1. Hexsel Cellulite Severity Scale

[0102]

[0103] The Clinician-Reported Photo-Digital Cellulite Severity Scale (CR-PCSS) is a validated 5-point photo-digital scale developed for investigators to use to rate the severity of cellulite in the participant's treatment area (e.g., one or both buttocks) through a live assessment. Ratings range from 0 (none) to 4 (severe) with labels and descriptors to aid the investigator in making the assessment. This assessment should be performed with the participant in a standing position with the gluteal muscles relaxed.

[0104] The Patient-Reported Photo-Digital Cellulite Severity Scale (PR-PCSS) is a validated 5-level photo-digital scale developed specifically for patients to rate the severity of cellulite in a treatment area (e.g., one or both buttocks) by viewing photographic digital images of each buttock taken by a camera. Ratings range from 0 (none) to 4 (severe) and are accompanied by images, labels, and descriptors to aid patients in their assessment.

[0105] The Investigator Global Aesthetic Improvement Scale (I-GAIS) is a 7-point scale ranging from 3 (very improved) to -3 (very worse) (Table 2). The investigator, who is a physician, will use the I-GAIS to determine the degree of improvement in each treatment area by comparing cellulite in pretreatment (baseline) images of each treatment area on Day 1 with images taken at the follow-up visit. The I-GAIS assessment will be based on digital photographs and will be performed separately for the 2 treatment areas.

[0106] Table 2. Investigator Global Aesthetic Improvement Scale (I-GAIS)

[0107]

[0108]

[0109] Subject and Investigator Cellulite Assessments - The Investigator cellulite assessments are independent of the subject assessments. Therefore, all subject cellulite assessments must be completed before the Investigator begins the cellulite assessment. Subject assessments will be conducted with the subject alone and without the presence of study site personnel in the room. Investigators are instructed not to verbalize their ratings in the presence of the subject and vice versa.

[0110] The Subject Global Aesthetic Improvement Scale (S-GAIS) is a 7-point scale ranging from 3 (very improved) to -3 (very worse). Subjects will use the S-GAIS to determine the extent of improvement in each treated area by comparing cellulite in the Day 1 pre-treatment (baseline) image of each treated area to the image taken at the follow-up visit.

[0111] The Body-Q assessment of cellulite is a subset of questions in the Body-Q questionnaire that measures a patient's perception of weight loss and / or body contouring.

[0112] The Investigator Satisfaction with Medication Administration Scale (Likert Scale) is a 5-point scale that can be used to determine the investigator's satisfaction with CCH (investigator's satisfaction with CCH administration).

[0113] Ultrasound Assessment - Surface ultrasound images of the treatment area can be captured. Pre-defined parameters can be measured / assessed from the images.

[0114] 3-D Photography - The PRIMOS 3-D camera can be used as an exploratory tool to objectively quantify the skin surface roughness score. The device and its associated imaging technology offer a new way to capture changes in skin laxity and the resolution of skin dimples caused by cellulite.

[0115] Subjects can be digitally photographed using the provided Canfield camera system while standing in a consistent, standard relaxed position with the gluteal muscles relaxed.

[0116] Treatment of cellulite may include improvement of at least one point on any of the above scales. Taking the CR-PCSS as an example, treatment of cellulite may include a severity rating from 4 to 3, a severity rating from 3 to 2, a severity rating from 2 to 1, a severity rating from 1 to 0, a severity rating from 4 to 2, a severity rating from 4 to 1, a severity rating from 4 to 0, etc.

[0117] In any of the methods disclosed herein, the subject can receive multiple treatment sessions. The subject can receive, for example, one, two, three, four, five, six, seven, eight, nine, ten, or more treatment sessions. In some embodiments, the treatment sessions are spaced 3 weeks (21 days). In some embodiments, the treatment sessions are spaced 6 weeks (42 days).

[0118] The amount of collagenase injected using the triple injection technique refers to the total amount of collagenase in three injection aliquots. The amount of collagenase injected using the single injection technique refers to the amount of collagenase in that single injection. The amount of collagenase administered per injection depends in part on the size of the treatment area, the severity of the cellulite, the injection technique (e.g., triple injection technique or single injection), and the volume administered per injection. For any of the methods disclosed herein, suitable amounts of collagenase administered to the treatment area per injection include, for example, about 0.0001 mg, about 0.001 mg, about 0.01 mg, about 0.02 mg, about 0.03 mg, about 0.035 mg, about 0.04 mg, about 0.05 mg, about 0.06 mg, about 0.07 mg, about 0.08 mg, about 0.09 mg, about 0.10 mg, about 0.20 mg, about 0.30 mg, about 0.40 mg, about 0.50 mg, about 0.60 mg, about 0.70 mg, about 0.80 mg, about 0.90 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 2.5 mg, about 3.0 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, or more than 5.0 mg. The amount of collagenase administered per injection can be about 0.0001 mg to about 5.0 mg, about 0.001 mg to about 5.0 mg, about 0.01 mg to about 5.0 mg, about 0.05 mg to about 5.0 mg, about 0.1 mg to about 5.0 mg, about 0.5 mg to about 5.0 mg, about 1.0 mg to about 5 mg, about 2.5 mg to about 5.0 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, or about 0.01 mg to about 0.1 mg. In some embodiments, about 0.028 mg of collagenase is administered per injection. In some embodiments, about 0.035 mg of collagenase is administered per injection. In some embodiments, about 0.07 mg of collagenase is administered per injection.

[0119] The amount of collagenase administered in a single treatment session depends in part on the size of the treatment area, the severity of the cellulite, the injection technique, the amount administered per injection, and the desired number of treatment sessions. For any of the disclosed methods, suitable amounts of collagenase administered to the treatment area in a single treatment session include, for example, about 0.01 mg, about 0.02 mg, about 0.03 mg, about 0.04 mg, about 0.05 mg, about 0.06 mg, about 0.07 mg, about 0.08 mg, about 0.09 mg, about 0.10 mg, about 0.20 mg, about 0.21 mg, about 0.30 mg, about 0.40 mg, about 0.42 mg, about 0.50 mg, about 0.60 mg, about 0.70 mg, about 0.80 mg, about 0.90 mg, about 100 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 5 0.80 mg, about 0.84 mg, about 0.90 mg, about 1.0 mg, about 2.0 mg, about 3.0 mg, about 4.0 mg, about 5.0 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10.0 mg, about 11.0 mg, about 12.0 mg, about 13.0 mg, about 14.0 mg, about 15.0 mg, about 16.0 mg, about 17.0 mg, about 18.0 mg, about 19.0 mg, about 20.0 mg or more. The amount of collagenase administered in a single treatment session can be from about 0.01 mg to about 20.0 mg, from about 0.01 mg to about 15.0 mg, from about 0.01 mg to about 10.0 mg, from about 0.01 mg to about 5.0 mg, from about 0.01 mg to about 1.0 mg, from about 0.01 mg to about 0.5 mg, from about 0.01 mg to about 0.05 mg, from about 0.05 mg to about 20 mg, from about 0.1 mg to about 20 mg, from about 0.5 mg to about 20 mg, from about 1.0 mg to about 20 mg, from about 2.5 mg to about 20 mg, from about 5.0 mg to about 20 mg, or from about 10 mg to about 20 mg. In some embodiments, about 0.21 mg to about 0.63 mg of collagenase is administered to the treatment area in a single treatment session. In some embodiments, about 0.27 mg to about 0.55 mg of collagenase is administered to the treatment area in a single treatment session. In some embodiments, about 0.21 mg of collagenase is applied to the treatment area in a single treatment session. In some embodiments, about 0.42 mg of collagenase is applied to the treatment area in a single treatment session. In some embodiments, about 0.84 mg of collagenase is applied to the treatment area in a single treatment session.

[0120] “Collagenase” means any of the following: (a) collagenase (including mutants) having activity as specified in EC 3.4.24.3 (https: / / www.brenda-enzymes.org / enzyme.php?ecno=3.4.24.3 (accessed on July 3, 2019); (b) Clostridium histolyticum (also known as Hathewaya histolytica); (c) CCH (as defined above); (d) a collagenase having at least 50% sequence alignment with collagenase I (also known as class I collagenase) as determined by BLAST; (e) a collagenase having at least 50% sequence alignment with collagenase II (also known as class || collagenase) as determined by BLAST; (f) a collagenase produced by fermentation of other organisms (i.e., non-Clostridium histolytica), such as mammalian, crustacean, fungal, bacterial or microbial origin; (g) a collagenase obtained by recombinant technology; (h) a collagenase having a molecular weight between about 65 kDa and about 130 kDa; (i) a collagenase designated as collagenase I (col I) or collagenase II (col II); (j) a mixture of collagenases I and II; (k) a collagenase from strain JCM 1403 (ATCC 19401) or a derivative thereof; (1) a collagenase from ATCC 21000 strain or a derivative thereof; (m) a collagenase from strain ATCC 69334 or a derivative thereof; (n) a collagenase from Clostridium perfringens; (o) a collagenase from Vibrio alginolyticus; (p) a collagenase from Streptomyces; (q) a collagenase from Pseudomonas; (r) a collagenase from Achromobacter, (s) a collagenase described by Worthington Biochem (www.worthington-biochem.com; “Product Highlights”); (t) a collagenase described by Sigma-Aldrich (www.sigma-aldrich.com); (u) a collagenase having one or more of the following characteristics:

[0121] ·V max (min -1 ) is about 0.08 to 7.70 (SRC assay; such as the assay described in International Publication No. WO2020 / 058755), or about 0.3 to 30.5 (GPA assay; such as the assay described in International Publication No. WO2020 / 058755);

[0122] ·K M , about 4.1 to 410 nanomolar (SRC assay), or, about 0.03 to 3.1 mM (GPA assay);

[0123] ·K cat (sec -1) is about 1.1 to about 107 (SRC determination), or, about 93 to 9179 (GPA determination);

[0124] 1 / K cat , microseconds of about 376 to 37222 (SRC determination), or, about 4 to 428 (GPA determination); or

[0125] ·K cat / K M (mM -1 sec -1 ), about 5140 to 508814 (SRC determination), or, about 60 to 5,934 (GPA determination);

[0126] (v) collagenase described by Nordmark Arzneimittel GmbH & Co. KG.; (w) collagenase from strain 004; (x) Biosimilars of the collagenase component of Biosimilars; (y) Biosimilars of collagenase components or or (z) an equivalent or mixture of any of the above. Non-limiting examples of collagenases that can be used in the present disclosure are described in U.S. Pat. No. 7,811,560, U.S. Pat. No. 9,757,435, U.S. Pat. No. 9,744,138, and International Publication No. WO2012 / 125948.

[0127] In some embodiments, the collagenase may include collagenase I. Suitable collagenase I include, for example, an amino acid sequence having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 1. In some aspects, collagenase I comprises the amino acid sequence of SEQ ID NO: 1.

[0128] In some embodiments, the collagenase may include collagenase II. Suitable collagenase II includes, for example, collagenase II comprising an amino acid sequence having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 2. In some aspects, collagenase II comprises the amino acid sequence of SEQ ID NO: 2.

[0129] Table 3. Sequence

[0130]

[0131]

[0132] In some embodiments, the collagenase may include a mixture of collagenase I and collagenase II. The collagenase may include, for example, a mixture of collagenase I and collagenase II, wherein collagenase I comprises an amino acid sequence having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 1, and collagenase II comprises an amino acid sequence having 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 2. In some aspects, the collagenase comprises a mixture of collagenase I and collagenase II, wherein collagenase I comprises the amino acid sequence of SEQ ID NO: 1, and collagenase II comprises the amino acid sequence of SEQ ID NO: 2. Suitable mixtures of collagenase I and collagenase II include, for example, a collagenase I: collagenase II mass ratio of 0.1:1, 0.25:1, 0.5:1, 0.75:1, 1:1, 1.1:1, 1.25:1, 1.5:1, 1.75:1, 2:1, 1:0.1, 1:0.25, 1:0.5; 1:0.75, 1:1.1, 1:1.25, 1:1.5, 1:1.75 or 1:2. Each of collagenase I and collagenase II may have a purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% as determined by, for example, reverse phase HPLC.

[0133] In some embodiments, the collagenase may include Clostridium histolyticum collagenase (CCH). As used herein, "CCH" refers to Clostridium histolyticum collagenase, which contains a mixture of collagenase I (SEQ ID NO: 1) and collagenase II (SEQ ID NO: 2) in a mass ratio of about 1: 1. CCH is obtained by fermentation of Clostridium histolyticum (also known as Hathewaya histolytica).

[0134] In some embodiments, collagenase I and collagenase II may have the following properties:

[0135] Collagenase I (SRC microplate assay)

[0136] οV max : Approximately 0.08 to 7.70 min -1

[0137] οK M : Approximately 4.1 to 410 nanomoles

[0138] οK cat : About 1.1 to 107 sec -1

[0139] ο1 / K cat : Approximately 376 to 37,222 microseconds

[0140] οK cat / K M : Approximately 5,140 to 508,814 mM -1 sec -1

[0141] Collagenase II (GPA microplate assay)

[0142] οV max :About 0.3About 30.5min -1

[0143] οK M : About 0.03 to 3.1mM

[0144] οK cat : Approximately 93 to 9,179 seconds -1

[0145] ο1 / K cat : Approximately 4 to 428 microseconds

[0146] οK cat / K M : Approximately 60 to 5,934 mM -1 sec -1

[0147] In some embodiments, collagenase I and collagenase II may have the following characteristics:

[0148] Collagenase I (SRC assay):

[0149] οV max : About 3.8min -1

[0150] οK M : Approximately 2.07x10 -4 mM

[0151] οK cat : About 53 seconds -1

[0152] ο1 / K cat : Approximately 18,799 microseconds

[0153] οk cat / K M : Approximately 256,977mM -1 sec -1

[0154] Collagenase II (GPA assay):

[0155] οV max : About 15.4min -1

[0156] οK M : About 1.6mM

[0157] οK cat : Approximately 4,636 seconds -1

[0158] ο1 / K cat : About 216 microseconds

[0159] οk cat / K M :About 2,997mM -1 sec -1

[0160] Preparations containing collagenase

[0161] Disclosed herein are formulations comprising: collagenase; lidocaine; and epinephrine.

[0162] Any of the collagenases described above can be used in the formulation. In some embodiments, the collagenase can include collagenase I. In some embodiments, the collagenase can include collagenase II. In some embodiments, the collagenase can include a mixture of collagenase I and collagenase II. In some aspects, the collagenase includes a mixture of collagenase I and collagenase II, wherein collagenase I comprises the amino acid sequence of SEQ ID NO: 1 and collagenase II comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the collagenase can include Clostridium histolyticum collagenase (CCH).

[0163] The concentration of collagenase can be about 0.23 mg / ml. The concentration of lidocaine can be about 5 mg / ml, about 10 mg / ml, about 15 mg / ml, or about 20 mg / ml. The concentration of epinephrine can be about 5 μg / ml or about 10 μg / ml. In some embodiments, the formulation can include about 0.23 mg / ml collagenase, about 2% lidocaine, and about 1:200,000 epinephrine.

[0164] The formulation may further comprise mannitol, sucrose and tromethamine. Thus, in some embodiments, the formulation comprises collagenase, lidocaine, epinephrine mannitol, sucrose and tromethamine.

[0165] Example

[0166] The following examples are provided to further describe some of the embodiments disclosed herein. These examples are intended to illustrate, but not to limit, the embodiments disclosed herein.

[0167] Example 1 - APHRODITE-1 Study

[0168] background

[0169] Clostridium histolyticum collagenase-aaes for injection (CCH-aaes; Endo Aesthetics LLC) is indicated for the treatment of moderate to severe cellulite in adult women on the buttocks. CCH-aaes injections are believed to initiate enzymatic excision and remodeling These include lysis of mature collagen-rich septa, resulting in the characteristic dimples of cellulite, rapid formation of new collagen, and some degree of fat lobule reorganization (formation of lobules of larger and more uniform size). In a pooled analysis of two randomized phase 3 trials of identical design in women with gluteal cellulite who received CCH-aaes (84%) or placebo (21%), the most common adverse event reported was injection site bruising. During these trials, 1 / 2-inch, three-part technique (also known as the three-injection technique, as described in the literature) was used. Figure 1 0.84 mg of CCH-aaes was injected into each treatment area (shown). Although injection site bruising following CCH-aaes administration typically resolves within 14 to 21 days, bruising is bothersome to some women and may be associated with subsequent skin discoloration. Therefore, a trial was designed to identify potential interventions that may reduce bruising and its associated consequences.

[0170] Purpose

[0171] Using a self-controlled study design, we evaluated the effects of several interventions for preventing or reducing buttock bruising in women with moderate to severe cellulite after CCH-AAES injections.

[0172] method

[0173] Study design and interventions

[0174] APHRODITE-1 (A Phase 2, open-label, self-controlled study of different interventions to reduce bruising following CCH treatment) is designed to enroll up to 150 women in the first phase of the trial. The trial is designed to evaluate different CCH-aaes doses, different CCH-aaes concentrations, different injection techniques (including injection depth), and diluent additives to prevent or reduce bruising at the injection site. The study is designed to add new intervention cohorts in later phases of the study as needed, while considering other diluent additives, alternative doses, and injection timing variables, including:

[0175] - Different CCH-aaes doses or different concentrations;

[0176] - Different injection techniques (including injection depth);

[0177] - contains a diluent additive; and / or

[0178] - Added new intervention queue.

[0179] Using an interactive response technology system, up to 30 participants (in a 1:1:1:1:1 ratio) were assigned to each of the eight cohorts, as shown in Table 4:

[0180] Table 4. Study intervention cohorts

[0181]

[0182]

[0183]

[0184] The patient population includes:

[0185] - Up to 180 women aged 18-60;

[0186] -Body mass index (BM) 18 to <32 kg / m 2 ;

[0187] - Moderate to severe cellulite on both buttocks (CR-PCSS grade 3 or 4);

[0188] -Hexsel CSS total score ≤ 12; and

[0189] - Fitzpatrick skin types I-IV.

[0190] Table 5 provides a summary of the demographic and baseline characteristics of those subjects who participated in the study.

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198] Injection site bruising (study and control sides) and other safety and efficacy parameters were monitored throughout the study. The trial included 4 follow-up visits after each CCH-aaes treatment period (days 1-2, 3-5, 6-9, and 10-14 after treatment) to assess the severity of injection site bruising and document the severity using standardized digital photographs ( Figure 4A and Figure 4B ).

[0199] Interim and ongoing assessments

[0200] At the time of submission of this application, study enrollment was complete but not all follow-up visits had occurred. Data generated from follow-up visits completed up to that date were assessed interim. Bruising severity will / has been determined by field assessment using the Investigator Assessment of the 5-point Bruising Severity Assessment Scale (IABSS), with each buttock assessed individually for comparison purposes ( Figure 2 The primary efficacy endpoint will be the proportion of participants who have / were IAB SS scores that were at least 1 grade lower in the left hip (study treatment) compared to the right hip (control treatment) 3-5 days after the first CCH-aaes injection at the Series 1 endpoint (second visit). Secondary efficacy endpoints include:

[0201] - the proportion of participants whose left hip IABSS score was at least 1 grade lower than the right hip at visit;

[0202] - Proportion of participants who improved ≥1 grade from baseline (Day 1) on the Investigator Global Aesthetic Improvement Scale (I-GAIS) by visit (I-GAIS uses a 7-point scale ranging from +3 ("very much improved") to -3 ("very much worse").

[0203] Each buttock was monitored throughout the study for adverse events, including injection site reactions.

[0204] in conclusion

[0205] APHRODITE-1 was designed to determine whether different interventions (i.e., dose, concentration, injection technique, and additional diluent additives) could prevent or reduce injection site bruising following CCH-aaes treatment of gluteal cellulite. The trial has the flexibility to add cohorts so that additional interventions can be tested over time.

[0206] The data provided below are special extracts of the data collected as of the time of submission of this application (before completing all subsequent visits).Table 6 shows the summary of the IABSS responders of the first level at the 2nd visit (1-2 days after treatment), the 3rd visit (3-5 days after treatment) and the 4th visit (6-9 days after treatment) of queues 1-6.For queues 7a and 7b, the equivalent visits (visit 2 and visit 7; visit 3 and visit 8; visit 4 and visit 9) of each buttock are listed.Although the data of the 90th day ± 7 days were collected, before and after the 4th visit, the degree of bruising on the study side (left buttock) and the control side (right buttock) after preliminary treatment began to subside, so that any visual difference of bruising between the two buttocks can be ignored.More precisely, before and after the 4th visit of queues 1-6 and the 4th / 9th visit (the corresponding visit of each buttock after initial treatment) of queues 7a and 7b, bruising began to subside. Therefore, only data for Visits 2, 3, and 4 (for Cohorts 1-6) and Visit 2 vs. Visit 7 / Visit 7 vs. Visit 2, Visit 3 vs. Visit 8 / Visit 8 vs. Visit 3, and Visit 4 vs. Visit 9 / Visit 9 vs. Visit 4 (for Cohorts 7a and 7b) are presented herein. As shown in Table 6, at Visit 3, more than 90% of subjects in Cohort 1 and more than 63% of subjects in Cohort 6 showed at least a 1-point improvement in IABSS. The percentage of subjects showing at least a 1-point improvement in IABSS at Visit 4 was lower than that at Visit 3 because, as described above, bruising had resolved on both the study and control sides by Visit 4.

[0207] Figure 5 Shown are the levels of bruising observed in patients in cohort 1 at visit 3 (days 3-5 after treatment). The left buttocks were injected with 0.42 mg of CCH-aaes at a concentration of 0.23 mg / mL using a three-injection technique, and the right buttocks (control) were injected with 0.84 mg of CCH-aaes at a concentration of 0.23 mg / mL using a three-injection technique. Figure 5 As shown, at least a 1-point improvement in IABSS was observed.

[0208] Table 6. Summary of Level 1 IABSS Respondents by Interview

[0209]

[0210]

[0211]

[0212]

[0213] Example U.S. Prescribing Information ( Label)

[0214] The following are approved by the U.S. Food and Drug Administration (FDA) A copy of the labeling that provides the U.S. prescribing information. The labeling was obtained from www_accessdata.fda.gov / drugsatfda_docs / label / 2020 / 761146s0001b1.pdf on May 10, 2023.

[0215]

[0216] Full Prescribing Information

[0217] 1 Indications and uses

[0218] QWO is indicated for the treatment of moderate to severe cellulite on the buttocks of adult women.

[0219] 2 Dosage and administration method

[0220] 2.1 Dosage

[0221] QWO is administered by subcutaneous injection at a dose of 0.84 mg per treatment area.

[0222] The treatment area was defined as a single buttock, with each treatment area receiving a maximum of 12 injections of QWO, 0.3 mL per injection (maximum total of 3.6 mL).

[0223] A treatment visit can include up to 2 treatment areas.

[0224] Treatment should be repeated every 21 days for a total of 3 treatment visits per treatment.

[0225] 2.2 Reconstitution of lyophilized powder

[0226] Before reconstitution, remove the vial from the refrigerator and allow it to sit at room temperature for at least 15 minutes. Inspect the vial containing QWO. The lyophilized powder cake should be white and intact with no signs of corrosion. The diluent should be a colorless solution with no particulate matter.

[0227] After removing the flip cap from the vial, use aseptic technique to wipe the rubber stopper and surrounding surfaces of the vial containing QWO and diluent with a sterile alcohol wipe (no other preservatives should be used).

[0228] Use only the provided diluent to reconstitute QWO.

[0229] Using an appropriately sized syringe and needle (not provided), withdraw the amount of diluent provided according to the number of injection sites (see Table 1).

[0230] Table 1: QWO reconfiguration instructions

[0231]

[0232] Slowly pour the diluent into the side of the vial containing the lyophilized powder of QWO. Do not invert the vial or shake the solution. Slowly swirl the solution to ensure that all the lyophilized powder has dissolved.

[0233] The reconstituted QWO solution in the vial may be stored at room temperature (20°C to 25°C / 68°F to 77°F) for up to 8 hours or refrigerated at 2°C to 8°C (36°F to 46°F) for up to 72 hours prior to administration. If the reconstituted QWO solution in the vial is refrigerated, bring the solution to room temperature for approximately 15 minutes before use.

[0234] The reconstituted QWO solution should be clear, colorless, and free of particulate matter. Visually inspect the solution for particulate matter or discoloration prior to administration. Do not inject if the reconstituted QWO solution is not clear, colorless, and substantially free of particulate matter.

[0235] Discard the syringe and needle used for reconstitution and the vial of diluent.

[0236] After reconstitution, the QWO solution in the vial can be used for only one injection session and for one patient only.

[0237] 2.3 Administration

[0238] Preparation of syringe for injection

[0239] Use a 1 mL syringe with a removable needle (not provided) and dispense 0.9 mL of reconstituted solution per syringe. Refer to Table 2 to prepare the appropriate number of syringes needed based on the number of injection sites. Once the syringes are ready, draw the remaining solution from the needle into the syringe and then replace the needle with a 30-gauge 1 / 2-inch needle. Use the reconstituted solution in the 1 mL syringe immediately. Do not store the reconstituted solution in the 1 mL syringe.

[0240] Table 2: QWO preparation instructions

[0241]

[0242] Injection technique

[0243] With the patient in a standing position, mark the injection site. With the patient in a prone position, inject QWO subcutaneously. Each QWO injection should be divided into three 0.1 mL aliquots (for a total injection volume of 0.3 mL) and injected at locations A, B, and C as shown in the figure below. The injection depth should be 0.5 inches (corresponding to the length of the needle) without applying downward pressure.

[0244]

[0245] Remove the needle completely from the skin and move to the next identified injection site. Each treatment area may receive up to 12 injections. After treatment, the patient should remain in the prone position for at least 5 minutes.

[0246] After you have completed your administration, do not store, pool, or use any vials or syringes containing unused reconstituted solution. Discard any unused portion.

[0247] 3 Dosage forms and specifications

[0248] For injection: 0.92 mg or 1.84 mg of Clostridium histolyticum collagenase, lyophilized powder (in the form of white cake), in single-dose vials.

[0249] 4 Contraindications

[0250] QWO is contraindicated in the following situations:

[0251] Patients with a history of hypersensitivity to collagenase or any of the excipients [see Warnings and Precautions (5.1)].

[0252] Infection at the injection site.

[0253] 5 Warnings and Precautions

[0254] 5.1 Hypersensitivity reactions

[0255] Serious hypersensitivity reactions, including anaphylaxis, have been reported with the use of C. histolytica collagenase. If such a reaction occurs, further injections of QWO should be discontinued and appropriate medical treatment instituted immediately.

[0256] 5.2 Injection Site Bruising

[0257] In clinical trials, 84% of subjects treated with QWO experienced injection site bruising [see Adverse Reactions (6.1)]. Subjects with coagulopathy or taking anticoagulant or antiplatelet medications (except those taking aspirin ≤ 150 mg daily) were excluded from Trials 1 and 2.

[0258] QWO should be used with caution in patients with bleeding abnormalities or in patients currently receiving antiplatelet therapy (except for patients taking aspirin ≤150 mg / day) or anticoagulant therapy.

[0259] 5.3 Alternatives to collagenase products

[0260] QWO should not be used as a substitute for other injectable collagenase products. QWO is not intended for the treatment of Peyronie's disease or Dupuytren's contracture.

[0261] 6 Adverse Reactions

[0262] The following adverse reactions for QWO for injection are described in more detail elsewhere in the labeling:

[0263] Hypersensitivity Reactions [see Contraindications (4) and Warnings and Precautions (5.1)].

[0264] Injection site bruising [see Warnings and Precautions (5.2)].

[0265] 6.1 Clinical Trial Experience

[0266] Because clinical trials are conducted under a wide variety of conditions, adverse reaction rates observed in the clinical trials of a drug cannot be directly compared to rates in the clinical trials of another drug and may not reflect the rates in clinical practice.

[0267] In two identically designed double-blind, placebo-controlled clinical trials (Trial 1 and Trial 2), 424 female subjects with cellulite on the buttocks were treated with QWO and 419 female subjects with cellulite were treated with placebo. The enrolled subjects were aged between 18 and 78 years, had moderate to severe cellulite (grade 3 or 4 on a scale of 0 to 4), and did not have excessive skin laxity. The majority of the subjects were white (78%) or African American (18%). In a separate open-label extension trial (Trial 3), subjects completed up to 3 treatment visits, separated by 21 days, and were followed up for up to 6 months after the last treatment visit.

[0268] Table 3 shows the incidence of adverse reactions that were reported by ≥1% of subjects receiving QWO and were more frequent than those receiving placebo in Trials 1 and 2 through Day 71. In general, adverse reactions did not last more than 21 days.

[0269] Table 3: Adverse Reactions Occurring in ≥1% of Subjects Throughout 71 Days in Trials 1 and 2

[0270]

[0271]

[0272] Merger Terms:

[0273] Bruising - injection site bruising, injection site hematoma, injection site hemorrhage (refers to the verbatim term injection site ecchymosis)

[0274] Pain – injection site pain, injection site discomfort, and injection site numbness

[0275] Swelling at the injection site, edema at the injection site, or hardening at the injection site

[0276] Discoloration - discoloration at the injection site

[0277] Nodules - injection site lumps and injection site nodules

[0278] A total of 479 participants from Trials 1 and 2 have completed a 6-month observation phase in the ongoing open-label safety extension (Trial 3). No long-term safety signals have been identified.

[0279] 6.2 Immunogenicity

[0280] As with all therapeutic proteins, there is the potential for immunogenicity. Improved detection of antibody production is dependent on the sensitivity and specificity of the assay. In addition, the observed incidence of antibody (including comprehensive antibodies) positivity in an assay may be influenced by multiple factors, including assay methodology, specimen handling, timing of specimen collection, concomitant medications, and underlying disease. For these reasons, comparison of antibody incidence in the studies described below with that in other studies or with other products, including other Clostridium histolyticum collagenase products, may be misleading.

[0281] By Day 22, anti-AUX-I antibodies and anti-AUX-II antibodies were found in approximately 53% (203 / 383) and 26% (101 / 383) of subjects, respectively, who completed the first treatment visit of QWO at the recommended dose in Trial 1 and Trial 2. The majority (>96%) of subjects developed antibodies to AUX-I and AUX-II after the second and third treatment visits. Antibody titers indicated that the antibodies were able to be maintained for up to 360 days after receiving the first recommended dose. By Day 71, approximately 68% and approximately 83% of subjects developed antibodies to AUX-I and AUX-II, respectively, which could be classified as neutralizing.

[0282] Antibodies to AUX-I and AUX-II, including those classified as neutralizing, were not associated with changes in clinical response or injection site adverse reactions.

[0283] 6.3 Post-marketing experience

[0284] Because adverse reactions are reported voluntarily from a population of uncertain size, it is not always possible to reliably estimate their frequency or establish a causal relationship to drug exposure. The following adverse reactions have been reported during post-approval use of collagenase products:

[0285] Immune System Disorders: Severe hypersensitivity reactions, including anaphylaxis [see Warnings and Precautions (5.1)].

[0286] 8 Use by Specific Groups

[0287] 8.1 Pregnancy

[0288] Risk Summary

[0289] There are no available data to evaluate the risk of drug-related major birth defects, miscarriage, or adverse maternal and fetal outcomes with the use of C. histolytica collagenase in pregnant women. Following subcutaneous administration, systemic concentrations of QWO are below the bioassay quantitation limit [see Clinical Pharmacology (12.3)].

[0290] In animal reproduction studies, no evidence of fetal harm was observed when Clostridium histolyticum collagenase was administered intravenously to pregnant rats during the period of organogenesis at doses up to 0.13 mg / rat (43 × the human equivalent dose [HED] on a mg / kg basis).

[0291] The estimated background risk of major birth defects and miscarriage in the specified population is unknown. In the U.S. general population, the estimated background risk of major birth defects and miscarriage in clinically recognized pregnancies is 2-4% and 15-20%, respectively.

[0292] 8.2 Breastfeeding

[0293] Risk Summary

[0294] There are no data regarding the effects of the presence of C. histolyticum collagenase, C. histolyticum, in human milk on the breastfed infant or on milk production. Following subcutaneous administration, systemic concentrations of QWO are below the bioassay limit of quantitation [see Clinical Pharmacology (12.3)]. The developmental and health benefits of breastfeeding should be considered alongside the mother's clinical need for C. histolyticum collagenase and any potential adverse effects on the breastfed child from C. histolyticum or the underlying maternal condition.

[0295] 8.4 Pediatric Use

[0296] The safety and effectiveness of QWO in pediatric patients have not been established.

[0297] 8.5 Elderly Use

[0298] In two double-blind, placebo-controlled clinical trials in subjects with cellulite (Trial 1 and Trial 2), 24 of the 424 subjects (5.7%) treated with QWO were 65 years of age or older. No overall differences in the safety of QWO were observed between elderly and younger patients.

[0299] 11Description

[0300] Clostridium histolyticum collagenase-aaes is a combination of bacterial collagenases AUX-I and AUX-II in a mass ratio of approximately 1:1, which are isolated and purified from the fermentation of Clostridium histolyticum bacteria.

[0301] Collagenase AUX-I is a single polypeptide chain consisting of approximately 1000 amino acids. Its observed molecular weight is 114 kilodaltons (kDa). It belongs to the class I Clostridium histolyticum collagenase.

[0302] Collagenase AUX-II is a single polypeptide chain consisting of approximately 1000 amino acids. Its observed molecular weight is 113 kilodaltons (kDa). It belongs to the class II Clostridium histolyticum collagenase.

[0303] QWO (Clostridium histolyticum collagenase) for injection is supplied as a sterile, preservative-free lyophilized powder (in the form of a white cake) in single-dose vials for subcutaneous use after reconstitution with QWO diluent.

[0304] Each single-dose vial of QWO 0.92-mg contains 0.92 mg of collagenase C. histolyticum-aaes and mannitol (37.7 mg), sucrose (18.9 mg), tromethamine (1.1 mg), and hydrochloric acid as needed to adjust the pH. Reconstitution with 4 mL of the supplied QWO diluent results in a solution with a pH of approximately 8.0 containing 0.23 mg / mL of C. histolyticum collagenase-aaes.

[0305] Each QWO 1.84-mg single-dose vial contains 1.84 mg of collagenase C. histolyticum-aaes and mannitol (75.4 mg), sucrose (37.8 mg), tromethamine (2.2 mg), and hydrochloric acid as needed to adjust pH. Reconstitution with 8 mL of the supplied QWO diluent results in a solution with a pH of approximately 8.0 and containing 0.23 mg / mL of C. histolyticum collagenase-aaes.

[0306] QWO Diluent is a sterile, preservative-free, colorless solution in single-dose vials containing 4 mL or 8 mL of 0.03% calcium chloride dihydrate and 0.6% sodium chloride and Water for Injection, USP.

[0307] 12 Clinical Pharmacology

[0308] 12.1 Mechanism of Action

[0309] Collagenase is a protease that hydrolyzes collagen in its native triple helical conformation under physiological conditions. The exact mechanism of action for the treatment of moderate to severe cellulite is unknown.

[0310] 12.2 Pharmacodynamics

[0311] The pharmacodynamics of QWO is still unclear.

[0312] 12.3 Pharmacokinetics

[0313] The pharmacokinetics of Clostridium histolyticum collagenase were evaluated in 140 female subjects with cellulite in four clinical trials. Plasma concentrations of Clostridium histolyticum collagenase type I (AUX-I) and Clostridium histolyticum collagenase type II were below the limit of quantification at 5 ng / mL and 25 ng / mL, respectively, in all subjects who received a single dose (QWO) of up to 3.36 mg in up to 4 treatment areas (up to 0.84 mg per treatment area).

[0314] 13 Nonclinical Toxicology

[0315] 13.1 Carcinogenesis, Mutagenesis, Impairment of Fertility

[0316] Long-term animal studies have not been conducted to evaluate the carcinogenic potential of Clostridium histolyticum collagenase.

[0317] Purified Clostridium histolyticum collagenase is not mutagenic to Salmonella typhimurium (Ames test) and is not clastogenic in both the in vivo mouse micronucleus test and the in vitro chromosome aberration test in human lymphocytes.

[0318] When administered intravenously to rats at doses up to 0.13 mg / rat (43×HED on a mg / kg basis), Clostridium histolyticum collagenase did not impair fertility and early embryonic development.

[0319] 14Clinical Research

[0320] Two randomized, multicenter, double-blind, placebo-controlled trials of identical design were conducted, Trial 1 and Trial 2, to evaluate the safety and efficacy of QWO in the treatment of cellulite in adult women. Eligible subjects had moderate (3) to severe (4) severity of cellulite on both buttocks, as assessed on a 5-point scale (0 = none; 4 = severe) by two subjects, using the patient-reported photo-digital cellulite severity scale (PR-PCSS), and by the investigator, using the clinician-reported photo-digital cellulite severity scale (CR-PCSS).

[0321] QWO was administered 0.84 mg per buttock as 12 subcutaneous injections (three 0.1 mL aliquots per injection, 0.3 mL per injection) for a total dose of 1.68 mg and a total volume of 7.2 mL (3.6 mL per buttock) per treatment visit. Three treatment visits were performed every 21 days.

[0322] In Trial 1 and Trial 2, the primary efficacy endpoint was the proportion of grade 2 multicomponent responders at day 71 after randomization. A grade 2 multicomponent responder was defined as an improvement of at least 2 grades from baseline in cellulite severity on the CR-PCSS and PR-PCSS of the target buttocks.

[0323] Patient satisfaction with the appearance of cellulite was assessed using a patient-reported outcome scale ranging from 0 (extremely dissatisfied) to 6 (extremely satisfied).

[0324] The mean age was 47 years and the mean BMI was 31 kg / m2. All subjects were female, and the majority were white (78%). At baseline, 61% of subjects had an investigator-reported cellulite severity (CR-PCSS) score of moderate, and 39% had a severe cellulite severity score.

[0325] Reductions in cellulite severity were observed more frequently in the QWO group compared with the placebo group, as measured by the investigator (CR-PCSS) and patient (PR-PCSS) scales at Day 71 (Table 4).

[0326] Table 4: Subject / Investigator 2-Grade Responder Analysis at Day 71

[0327]

[0328]

[0329] Adj Trt Diff = adjusted treatment difference; CI = confidence interval

[0330] Non-responder imputation was used to handle missing data. The adjusted treatment difference was the weighted average of the treatment differences in percent response across analysis centers using Cochran-Mantel-Haenszel (CMH) weights and associated CIs.

[0331] In both Trials 1 and 2, patient-reported measures of satisfaction with the appearance of cellulite showed greater improvements in the QWO group than in the placebo group.

[0332] 16 How to supply / Storage and handling

[0333] How to Supply

[0334] QWO (Collagenase Clostridium histolyticum-aaes) for injection is a sterile, preservative-free, lyophilized powder (in the form of a white cake) for subcutaneous administration in a single-dose vial.

[0335]

[0336] Storage and processing

[0337] Refrigerate QWO and diluents for vials of QWO at 2°C to 8°C (36°F to 46°F). Refrigeration is not required.

[0338] 17 Patient Counseling Information

[0339] Inform patients to read the FDA-approved patient labeling (Patient Information)

[0340] Hypersensitivity reactions

[0341] Inform patients to seek medical attention immediately if they experience any of the above symptoms of a serious hypersensitivity reaction [see Warnings and Precautions (5.1)].

[0342] Bruising at the injection site

[0343] Inform patients that injection site bruising may occur after taking QWO [see Warnings and Precautions (5.2)].

[0344] Manufacturer:

[0345] Endo Global Aesthetics Limited

[0346] Dublin, Ireland

[0347] US License Number: 2136

[0348] Distributor:

[0349] Endo Aesthetics LLC

[0350] Malvern, Pennsylvania 19355

[0351]

[0352]

[0353] This patient information has been approved by the U.S. Food and Drug Administration (FDA) Release Date: July 2020

[0354] Those skilled in the art will appreciate that various changes and modifications may be made to the preferred embodiments disclosed herein, and these changes and modifications may not depart from the spirit of the invention. Therefore, the appended claims are intended to cover all equivalent variations that fall within the spirit and scope of the invention.

[0355] The disclosures of each patent, patent application, and publication cited or described in this document are incorporated herein by reference in their entirety.

[0356] Implementation

[0357] The following list of embodiments is intended to supplement, not replace or supersede, what was described previously.

[0358] Embodiment 1. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0359] A total dose of about 0.42 mg of collagenase is subcutaneously administered to the treatment area of ​​the subject per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0360] Embodiment 2. The method according to embodiment 1, wherein the collagenase is administered using a three-injection technique.

[0361] Embodiment 3. The method of embodiment 1 or 2, wherein the collagenase is applied at a depth of about 0.5 inches.

[0362] Embodiment 4. A method according to any one of embodiments 1-3, wherein the bruising is reduced compared to the level of bruising associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase to the treatment area of ​​the subject using a three-injection technique at a depth of about 0.5 inches per treatment session.

[0363] Embodiment 5. The method of any one of embodiments 1-4, wherein the collagenase has a concentration of approximately 0.23 mg / ml.

[0364] Embodiment 6. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0365] A total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml is subcutaneously administered to the treatment area of ​​the subject during each treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0366] Embodiment 7. The method according to embodiment 6, wherein the collagenase is administered using a three-injection technique.

[0367] Embodiment 8. The method of embodiment 6 or 7, wherein the collagenase is applied at a depth of about 0.5 inches.

[0368] Embodiment 9. The method of any one of embodiments 6-8, wherein the bruising is reduced compared to the level of bruising associated with subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of ​​the subject using a three-injection technique at a depth of about 0.5 inches per treatment session, wherein the collagenase concentration is about 0.23 mg / ml.

[0369] Embodiment 10. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0370] A total dose of about 0.84 mg of collagenase is subcutaneously administered to the treatment area of ​​the subject at a depth of about 0.25 inches per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0371] Embodiment 11. The method according to embodiment 10, wherein the collagenase is administered using a three-injection technique.

[0372] Embodiment 12. The method of embodiment 10 or 11, wherein the bruising is reduced compared to the level of bruising associated with administering a total dose of about 0.84 mg of collagenase subcutaneously to the treatment area of ​​the subject using a three-injection technique at a depth of about 0.5 inches per treatment session.

[0373] Embodiment 13. The method of any one of embodiments 10-12, wherein the collagenase has a concentration of approximately 0.23 mg / ml.

[0374] Embodiment 14. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0375] A total dose of about 0.84 mg of collagenase having a concentration of about 0.09 mg / ml is subcutaneously administered to the treatment area of ​​the subject during each treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0376] Embodiment 15. The method of embodiment 14, wherein the collagenase is administered using a single aliquot injection.

[0377] Embodiment 16. The method of embodiment 14 or 15, wherein the collagenase is applied at a depth of about 0.25 inches.

[0378] Embodiment 17. The method of any one of embodiments 14-16, wherein the administration comprises up to 30 single aliquot injections of collagenase per treatment period.

[0379] Embodiment 18. The method of any one of embodiments 14-17, wherein the collagenase has a concentration of approximately 0.23 mg / mL and bruising is reduced compared to the level of bruising associated with subcutaneous administration of a total dose of approximately 0.84 mg of collagenase to the treatment area of ​​the subject using a single aliquot injection technique at a depth of approximately 0.25 inches per treatment session.

[0380] Embodiment 19. A method according to any one of embodiments 14-18, wherein said administration comprises up to 12 single aliquot injections of collagenase per treatment period.

[0381] Embodiment 20. The method of any one of embodiments 14-19, wherein the injections are spaced about 2 cm to about 3 cm apart.

[0382] Embodiment 21. A method according to embodiment 20, wherein the injections are spaced apart in a grid pattern.

[0383] Embodiment 22. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0384] A composition comprising collagenase, lidocaine, and epinephrine is subcutaneously administered to the treatment area of ​​the subject, wherein a total dose of approximately 0.42 mg of collagenase is administered using a three-injection technique per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0385] Embodiment 23. A method according to embodiment 22, wherein the collagenase is administered using a three-injection technique.

[0386] Embodiment 24. The method of embodiment 22 or 23, wherein the collagenase is applied at a depth of about 0.5 inches.

[0387] Embodiment 25. The method of any one of embodiments 22-24, wherein the bruising is reduced compared to the level of bruising associated with subcutaneous administration of a collagenase to the treatment area of ​​the subject, wherein a total dose of approximately 0.84 mg of collagenase is administered at a depth of approximately 0.5 inches using a three-injection technique in each treatment session.

[0388] Embodiment 26. The method of any one of embodiments 22-25, wherein the collagenase has a concentration of approximately 0.23 mg / ml.

[0389] Embodiment 27. A method according to any of the preceding embodiments, wherein the subject receives multiple treatment stages.

[0390] Embodiment 28. A method according to embodiment 27, wherein the treatment phases are spaced 3 weeks apart.

[0391] Embodiment 29. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0392] A total dose of about 0.21 mg of collagenase having a concentration of about 0.12 mg / mL is subcutaneously administered to the treatment area of ​​the subject during each treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0393] Embodiment 30. The method of embodiment 29, wherein the collagenase is administered using a three-injection technique.

[0394] Embodiment 31. The method of embodiment 29 or 30, wherein the collagenase is applied at a depth of about 0.5 inches.

[0395] Embodiment 32. The method of any one of embodiments 29-31, wherein the bruising is reduced compared to the level of bruising associated with subcutaneously administering a total dose of about 0.42 mg of collagenase to the treatment area of ​​the subject during each treatment period, wherein the collagenase has a concentration of about 0.23 mg / ml and is injected using a three-injection technique at a depth of about 0.5 inches.

[0396] Embodiment 33. A method according to any one of embodiments 29-32, wherein the subject receives more than one treatment period, and the treatment periods are separated by approximately 42 days.

[0397] Embodiment 34. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising:

[0398] Tranexamic acid is orally administered to the subject during each treatment period, and a total dose of approximately 0.84 mg of collagenase is subcutaneously administered to the subject's treatment area, thereby reducing collagenase-mediated bruising in the subject.

[0399] Embodiment 35. A method according to embodiment 34, wherein the collagenase is administered using a three-injection technique.

[0400] Embodiment 36. The method of embodiment 34 or 35, wherein the collagenase is applied at a depth of about 0.5 inches.

[0401] Embodiment 37. The method of any one of Embodiments 34-36, comprising administering to the subject about 500 mg to about 1500 mg of tranexamic acid.

[0402] Embodiment 38. The method of embodiment 37, comprising administering approximately 1300 mg of tranexamic acid to the subject.

[0403] Embodiment 39. The method according to any of the preceding embodiments, wherein the collagenase is administered as a composition comprising collagenase, mannitol, sucrose and tromethamine.

[0404] Embodiment 40. The method according to any of the preceding embodiments, wherein the assessment of bruise grade is an investigator-assessed bruise severity scale (IABSS).

[0405] Embodiment 41. A method of treating cellulite in a subject, the method comprising:

[0406] a) treating cellulite in the subject by subcutaneously administering a total dose of about 0.42 mg of collagenase to the treatment area of ​​the subject per treatment session;

[0407] b) subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml per treatment session, thereby treating cellulite in the subject;

[0408] c) subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.23 mg / ml to a depth of about 0.25 inches per treatment session, thereby treating cellulite in the subject;

[0409] d) subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.09 mg / ml per treatment session, thereby treating cellulite in the subject;

[0410] e) subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to the treatment area of ​​the subject, wherein a total amount of about 0.42 mg of collagenase is administered per treatment session, thereby treating cellulite in the subject;

[0411] f) subcutaneously administering to the treatment area of ​​the subject a total amount of about 0.21 mg of collagenase having a concentration of about 0.12 mg / ml per treatment session, thereby treating cellulite in the subject; or

[0412] g) treating cellulite in the subject by orally administering tranexamic acid to the subject and subcutaneously administering a total amount of approximately 0.84 mg of collagenase to the subject's treatment area during each treatment period.

[0413] Embodiment 42. A method according to embodiment 41, wherein the collagenase is administered using a three-injection technique.

[0414] Embodiment 43. A method according to embodiment 41, wherein the collagenase is administered using a single aliquot injection.

[0415] Embodiment 44. The method of embodiment 42 or 43, wherein the collagenase is applied at a depth of about 0.25 inches or about 0.5 inches.

[0416] Embodiment 45. The method of any one of embodiments 41-44, wherein the tranexamic acid in step g) has a concentration of about 500 mg to about 1500 mg.

[0417] Embodiment 46. The method of embodiment 45, wherein the tranexamic acid in step g) has a concentration of about 1300 mg.

[0418] Embodiment 47. The method of any one of embodiments 41-46, wherein the collagenase is administered as a composition comprising collagenase, mannitol, sucrose, and tromethamine.

[0419] Embodiment 48. The method of embodiment 41, wherein in part a) the collagenase has a concentration of about 0.23 mg / ml.

[0420] Embodiment 49. The method of embodiment 41, wherein the injections in part d) are spaced about 2 cm to about 3 cm apart.

[0421] Embodiment 50. A method according to embodiment 49, wherein the injections are spaced apart in a grid pattern.

[0422] Embodiment 51. A method according to any one of embodiments 41-50, wherein the subject receives multiple treatment stages.

[0423] Embodiment 52. A method according to embodiment 51, wherein the treatment phases are spaced 3 weeks apart.

[0424] Embodiment 53. The method of embodiment 51, wherein the treatment phases are approximately 42 days apart.

[0425] Embodiment 54. A method according to any of embodiments 41-53, wherein the treatment can be assessed by a scale or measurement tool selected from the following: Hexsel Cellulite Severity Grading Scale (HexselCSS), Hexsel Dimple Depth Rating Scale, Likert Scale, Dimple Analysis, Clinician-Reported Photo-Digital Cellulite Severity Scale (CR-PCSS), Patient-Reported Photo-Digital Cellulite Severity Scale (PR-PCSS), Investigator Global Aesthetic Improvement Scale (I-GAIS), Subject Global Aesthetic Improvement Scale (S-GAIS), Patient-Reported Cellulite Impact Scale (PR-CIS), PR-CIS Simplified, Patient Self-Rating Scale (SSRS), Subject Satisfaction with Cellulite Treatment Questionnaire (SSCT), Clinician Assessment of Cellulite Severity (Photographic or Other Images), Body-Q, and a validated image-digital or other clinical and / or patient-used scale for evaluating cellulite severity, improvement, and / or patient satisfaction.

[0426] Embodiment 55. A method according to any one of the preceding embodiments, wherein the collagenase comprises collagenase I, collagenase II, or a mixture of collagenase I and collagenase II.

[0427] Embodiment 56. A method according to embodiment 55, wherein the collagenase comprises a mixture of collagenase I and collagenase II.

[0428] Embodiment 57. A method according to embodiment 56, wherein the collagenase I comprises the amino acid sequence of SEQ ID NO: 1, and the collagenase II comprises the amino acid sequence of SEQ ID NO: 2.

[0429] Embodiment 58. The method according to any of the preceding embodiments, wherein the collagenase is Clostridium histolyticum collagenase (CCH).

[0430] Embodiment 59. A method according to any of the preceding embodiments, wherein the treatment area is the left buttock, the right buttock, or both the left buttock and the right buttock.

[0431] Embodiment 60. A formulation comprising:

[0432] A collagenase;

[0433] Lidocaine; and

[0434] Adrenaline.

[0435] Embodiment 61. The formulation according to embodiment 60, comprising:

[0436] About 0.23 mg / mL of collagenase;

[0437] approximately 2% lidocaine; and

[0438] About 1:200,000 of adrenaline.

[0439] Embodiment 62. The preparation of embodiment 60 or 61, wherein the collagenase comprises collagenase I, collagenase II, or a mixture of collagenase I and collagenase II.

[0440] Embodiment 63. A formulation according to embodiment 62, wherein the collagenase comprises a mixture of collagenase I and collagenase II.

[0441] Embodiment 64. The formulation of embodiment 63, wherein the collagenase I comprises the amino acid sequence of SEQ ID NO: 1, and the collagenase II comprises the amino acid sequence of SEQ ID NO: 2.

[0442] Embodiment 65. The formulation of any one of embodiments 60-64, wherein the collagenase is Clostridium histolyticum collagenase (CCH).

Claims

1. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising: A total dose of about 0.42 mg of collagenase is subcutaneously administered to the treatment area of ​​the subject per treatment session, thereby reducing collagenase-mediated bruising in the subject.

2. The method of claim 1, wherein the collagenase is administered using a three-injection technique.

3. The method of claim 1 or 2, wherein the collagenase is administered at a depth of about 0.5 inches.

4. The method of any one of claims 1-3, wherein the collagenase has a concentration of about 0.23 mg / ml.

5. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising: A total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml is subcutaneously administered to the treatment area of ​​the subject during each treatment session, thereby reducing collagenase-mediated bruising in the subject.

6. The method of claim 5, wherein the collagenase is administered using a three-injection technique.

7. The method of claim 5 or 6, wherein the collagenase is administered at a depth of about 0.5 inches.

8. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising: A total dose of about 0.84 mg of collagenase is subcutaneously administered to the treatment area of ​​the subject at a depth of about 0.25 inches per treatment session, thereby reducing collagenase-mediated bruising in the subject.

9. The method of claim 8, wherein the collagenase is administered using a three-injection technique.

10. The method of claim 8 or 9, wherein the collagenase has a concentration of about 0.23 mg / ml.

11. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising: A total dose of about 0.84 mg of collagenase having a concentration of about 0.09 mg / ml is subcutaneously administered to the treatment area of ​​the subject during each treatment session, thereby reducing collagenase-mediated bruising in the subject.

12. The method of claim 11, wherein the collagenase is administered using a single aliquot injection.

13. The method of claim 11 or 12, wherein the collagenase is administered at a depth of about 0.25 inches.

14. The method of any one of claims 11-13, wherein the administration comprises up to 30 single aliquot injections of collagenase per treatment period.

15. The method of any one of claims 11-13, wherein said administering comprises up to 12 single aliquot injections of collagenase per treatment period.

16. The method of any one of claims 11-15, wherein the injections are spaced about 2 cm to about 3 cm apart.

17. The method of any one of claims 11-16, wherein the injections are spaced apart in a grid pattern.

18. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising: A composition comprising collagenase, lidocaine, and epinephrine is subcutaneously administered to the treatment area of ​​the subject, wherein a total dose of approximately 0.42 mg of collagenase is administered using a three-injection technique per treatment session, thereby reducing collagenase-mediated bruising in the subject.

19. The method of claim 18, wherein the collagenase is administered using a three-injection technique.

20. The method of claim 18 or 19, wherein the collagenase is administered at a depth of about 0.5 inches.

21. The method of any one of claims 18-20, wherein the collagenase has a concentration of about 0.23 mg / ml.

22. The method according to any one of the preceding claims, wherein the subject receives multiple treatment sessions.

23. The method of claim 22, wherein the treatment phases are spaced 3 weeks apart.

24. A method of reducing collagenase-mediated bruising in a subject having cellulite, the method comprising: A total dose of about 0.21 mg of collagenase having a concentration of about 0.12 mg / mL is subcutaneously administered to the treatment area of ​​the subject during each treatment session, thereby reducing collagenase-mediated bruising in the subject.

25. The method of claim 24, wherein the collagenase is administered using a three-injection technique.

26. The method of claim 24 or 25, wherein the collagenase is administered at a depth of about 0.5 inches.

27. The method of any one of claims 24-26, wherein the subject receives more than one treatment period, the treatment periods being approximately 42 days apart.

28. The method of any one of the preceding claims, wherein the collagenase is administered as a composition comprising collagenase, mannitol, sucrose and tromethamine.

29. The method according to any one of the preceding claims, wherein the assessment of bruise grade is an investigator assessed bruise severity scale (IABSS).

30. A method of treating cellulite in a subject, the method comprising: a) treating cellulite in the subject by subcutaneously administering a total dose of about 0.42 mg of collagenase to the treatment area of ​​the subject per treatment session; b) subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.05 mg / ml per treatment session, thereby treating cellulite in the subject; c) subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.23 mg / ml to a depth of about 0.25 inches per treatment session, thereby treating cellulite in the subject; d) subcutaneously administering to the treatment area of ​​the subject a total dose of about 0.84 mg of collagenase having a concentration of about 0.09 mg / ml per treatment session, thereby treating cellulite in the subject; e) subcutaneously administering a composition comprising collagenase, lidocaine, and epinephrine to the treatment area of ​​the subject, wherein a total amount of about 0.42 mg of collagenase is administered per treatment session, thereby treating cellulite in the subject; f) subcutaneously administering a total amount of about 0.21 mg of collagenase having a concentration of about 0.12 mg / ml to the treatment area of ​​the subject per treatment session, thereby treating cellulite in the subject.

31. The method of claim 30, wherein the collagenase is administered using a three-injection technique.

32. The method of claim 30, wherein the collagenase is administered using a single aliquot injection.

33. The method of any one of claims 30-32, wherein the collagenase is applied at a depth of about 0.25 inches or about 0.5 inches.

34. The method of any one of claims 30-33, wherein the collagenase is administered as a composition comprising collagenase, mannitol, sucrose and tromethamine.

35. The method of claim 30, wherein in part a) the collagenase has a concentration of about 0.23 mg / ml.

36. The method of claim 30, wherein the injections in portion d) are spaced about 2 cm to about 3 cm apart.

37. The method of claim 36, wherein the injections are spaced apart in a grid pattern.

38. The method of any one of claims 30-37, wherein the subject receives multiple treatment sessions.

39. The method of claim 38, wherein the treatment phases are spaced 3 weeks apart.

40. The method of claim 38, wherein the treatment phases are approximately 42 days apart.

41. The method of any one of claims 30-40, wherein the treatment can be assessed by a scale or measurement tool selected from the group consisting of: Hexsel Cellulite Severity Grading Scale (HexselCSS), Hexsel Dimple Depth Rating Scale, Likert Scale, Dimple Analysis, Clinician-Reported Photo-Digital Cellulite Severity Scale (CR-PCSS), Patient-Reported Photo-Digital Cellulite Severity Scale (PR-PCSS), Investigator Global Aesthetic Improvement Scale (I-GAIS), Subject Global Aesthetic Improvement Scale (S-GAIS), Patient-Reported Cellulite Impact Scale (PR-CIS), PR-CIS Simplified, Patient Self-Rating Scale (SSRS), Subject Satisfaction with Cellulite Treatment Questionnaire (SSCT), Clinician Assessment of Cellulite Severity (Photographic or Other Images), Body-Q, and a validated image-digital or other clinical and / or patient-used scale for assessing cellulite severity, improvement, and / or patient satisfaction.

42. The method of any of the preceding claims, wherein the collagenase comprises collagenase I, collagenase II, or a mixture of collagenase I and collagenase II.

43. The method of claim 42, wherein the collagenase comprises a mixture of collagenase I and collagenase II.

44. The method of claim 43, wherein the collagenase I comprises the amino acid sequence of SEQ ID NO: 1, and the collagenase II comprises the amino acid sequence of SEQ ID NO:

2.

45. The method of any one of the preceding claims, wherein the collagenase is Clostridium histolyticum collagenase (CCH).

46. ​​The method of any of the preceding claims, wherein the treatment area is the left buttock, the right buttock, or both the left buttock and the right buttock.

47. A preparation comprising: A collagenase; Lidocaine; as well as Adrenaline.

48. The formulation of claim 47, comprising: About 0.23 mg / mL of collagenase; About 2% lidocaine; as well as About 1:200,000 of adrenaline.

49. The preparation of claim 47 or 48, wherein the collagenase comprises collagenase I, collagenase II, or a mixture of collagenase I and collagenase II.

50. The preparation of claim 49, wherein the collagenase comprises a mixture of collagenase I and collagenase II.

51. The preparation of claim 50, wherein the collagenase I comprises the amino acid sequence of SEQ ID NO: 1 and the collagenase II comprises the amino acid sequence of SEQ ID NO:

2.

52. The preparation of any one of claims 47-51, wherein the collagenase is Clostridium histolyticum collagenase (CCH).

Images