Method for reducing collagenase-mediated abrasions in a subject having cellulite

Abstract

Methods for reducing collagenase-mediated bruising in a subject having cellulite, methods for treating cellulite in a subject, and collagenase-containing formulations are disclosed herein.

Description

Technical Field

[0001] Cross - reference to related applications This application claims priority to U.S. Provisional Application No. 63 / 378,424, filed on October 5, 2022, and U.S. Provisional Application No. 63 / 340,295, filed on May 10, 2022, the disclosures of each of which are hereby incorporated by reference in their entirety.

[0002] Technical Field Disclosed herein are methods for reducing collagenase - mediated bruising in a subject having cellulite, methods for treating cellulite in a subject, and collagenase - containing formulations.

Background Art

[0003] Background Collagenase is a protease that hydrolyzes collagen in its native triple - helix conformation under physiological conditions. QWO® (CCH - aaes), a combination of AUX - I and AUX - II (also referred to as collagenase I and collagenase II), is approved for the treatment of moderate to severe cellulite in the buttocks of adult women. QWO® effectively lyses the subcutaneous fibrous septa that are the cause of skin dimpling in women with cellulite at the injection site. QWO® exhibits an acceptable safety and immunogenicity profile, and injection - site bruising is the most common adverse event. Bruising associated with QWO® generally resolves within 21 days and before the next treatment session, but bruising is bothersome to participants due to the potential for skin discoloration as well as associated swelling and pain.

Summary of the Invention

[0004] Summary A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.42 mg per treatment session, thereby reducing collagenase-mediated bruising in the subject.

[0005] A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing collagenase-mediated bruising in the subject.

[0006] A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session at a depth of about 0.25 inches, thereby reducing collagenase-mediated bruising in the subject.

[0007] A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing collagenase-mediated bruising in the subject.

[0008] A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine and epinephrine, wherein a total dose of collagenase of about 0.42 mg per treatment session is administered, thereby reducing collagenase-mediated bruising in the subject.

[0009] A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.21 mg per treatment session, the collagenase having a concentration of about 0.12 mg / ml, thereby reducing collagenase-mediated bruising in the subject.

[0010] A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein, the method comprising orally administering tranexamic acid (TXA) to the subject per treatment session and subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg, thereby reducing collagenase-mediated bruising in the subject.

[0011] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.42 mg per treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0012] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0013] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject about 0.84 mg total dose of collagenase at a depth of about 0.25 inches per treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0014] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject about 0.84 mg total dose of collagenase having a concentration of about 0.09 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0015] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine and epinephrine, wherein about 0.42 mg total dose of collagenase is administered per treatment session, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0016] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein, the method comprising subcutaneously administering to a treatment area of the subject about 0.21 mg total dose of collagenase having a concentration of about 0.12 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0017] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein. The method includes orally administering tranexamic acid to the subject per treatment session and subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of the subject, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject.

[0018] A method for treating cellulite in a subject is also provided. The method includes subcutaneously administering a total dose of about 0.42 mg of collagenase per treatment session to the treatment area of the subject, thereby treating cellulite in the subject.

[0019] A method for treating cellulite in a subject is disclosed herein. The method includes subcutaneously administering a total dose of about 0.84 mg of collagenase per treatment session to the treatment area of the subject, the collagenase having a concentration of about 0.05 mg / ml, thereby treating cellulite in the subject.

[0020] A method for treating cellulite in a subject is disclosed herein. The method includes subcutaneously administering a total dose of about 0.84 mg of collagenase per treatment session to the treatment area of the subject at a depth of about 0.25 inches, the collagenase having a concentration of about 0.23 mg / ml, thereby treating cellulite in the subject.

[0021] A method for treating cellulite in a subject is disclosed herein. The method includes subcutaneously administering a total dose of about 0.84 mg of collagenase per treatment session to the treatment area of the subject, the collagenase having a concentration of about 0.09 mg / ml, thereby treating cellulite in the subject.

[0022] A method for treating cellulite in a subject is disclosed herein, the method comprising the step of subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine and epinephrine, wherein a total dose of about 0.42 mg of collagenase is administered per treatment session, thereby treating cellulite in the subject.

[0023] A method for treating cellulite in a subject is disclosed herein, the method comprising the step of subcutaneously administering to a treatment area of the subject a total dose of about 0.21 mg of collagenase per treatment session, the collagenase having a concentration of about 0.12 mg / ml, thereby treating cellulite in the subject.

[0024] Also provided herein is a formulation comprising collagenase, lidocaine and epinephrine.

Brief Description of the Drawings

[0025] Brief Description of the Drawings The Summary and the following Detailed Description will be better understood when read in conjunction with the accompanying Drawings. For purposes of illustrating the disclosed methods and formulations, exemplary embodiments of the methods and formulations are shown in the Drawings, but the methods and formulations are not limited to the specific embodiments disclosed. In the Drawings:

Figure 1

Figure 2

Figure 3

Figure 4A

Figure 4B

Figure 5

[0026] Detailed Description of Exemplary Embodiments The disclosed methods and formulations can be more readily understood by reference to the following detailed description, which forms a part of the present disclosure and is to be read in conjunction with the accompanying drawings. The disclosed methods and formulations are not limited to the specific methods and formulations described and / or shown herein, and the terms used herein are for the purpose of describing particular embodiments by way of example only and are not intended to limit the claimed methods and formulations.

[0027] Unless otherwise specifically stated, any description of a possible mechanism or form of action or reason for improvement is illustrative only, and the disclosed methods and formulations are not restricted by the accuracy or inaccuracy of any such suggested mechanism or form of action or reason for improvement.

[0028] When a numerical range is recited or established herein, the range includes its endpoints and all individual integers and rational numbers within the range, and each of the narrower ranges formed by all the various possible combinations of these endpoints as well as the internal integers and rational numbers is also included, to the extent that each of these narrower ranges is described as clearly as if it were specifically recited. When a numerical range is described herein as being greater than a recited value, nevertheless the range is finite and is bounded at its upper end by values that are operable within the context of the disclosure herein. When a numerical range is described herein as being less than a recited value, nevertheless the range is bounded at its lower end by a value that is not zero. When defining a range, it is not intended that the ranges of methods and formulations be limited to the specific values recited. All ranges are inclusive and combinable.

[0029] It is understood that specific features of the disclosed methods and formulations described herein in the context of separate aspects may also be provided in combination in a single aspect. Conversely, various features of the disclosed methods and formulations described herein in the context of a single aspect may also be provided separately or in any sub-combination.

[0030] As used herein, the singular forms "a", "an" and "the" include the plural.

[0031] Various terms relating to the described aspects are used throughout the specification and claims. Such terms are given their ordinary meaning in the relevant art unless otherwise indicated. Other specifically defined terms are intended in a manner consistent with the definitions provided herein.

[0032] When the term "about" is used in relation to a numerical range, the cutoff or specific value is used to indicate that the recited value can vary by up to about 15% from the recited value. Thus, the term "about" is used to encompass variations of ±15% or less, ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from a particular value. When a value is presented as an approximation, it is understood that the use of the antecedent "about" forms a separate aspect. A reference to a particular numerical value includes at least that particular value unless the context clearly dictates otherwise.

[0033] The term "comprising" is intended to include examples encompassed by the terms "consisting essentially of" and "consisting of"; similarly, the term "consisting essentially of" is intended to include examples encompassed by the term "consisting of".

[0034] As used herein, the term "subject" is intended to mean any animal, particularly a mammal. Thus, the methods are applicable to humans and non-human animals, but most preferably to humans. "Subject" and "patient" are used interchangeably herein.

[0035] As used herein, "administering" and like terms refer to the procedure by which collagenase or a composition / pharmaceutical formulation comprising collagenase is injected into a subject such that a target cell, tissue, or compartment of the subject's body is contacted with the collagenase.

[0036] As used herein, "collagenase-mediated bruising" refers to at least one of bruising, hematoma, bleeding, petechiae, and discoloration occurring at and / or around the injection site after administration of collagenase.

[0037] "Treat", "treatment", and similar terms include reducing and / or eliminating cellulite, reducing and / or eliminating the causes underlying cellulite, reducing and / or eliminating the potential for cellulite, and inducing and / or improving the overall cosmetic quality of the skin.

[0038] As used herein, "reducing bruising" and similar phrases refer to at least a 1-level improvement on the investigator assessment of the bruising severity scale (IABSS).

[0039] The term "treatment area" refers to the area of a subject having cellulite to which a specific total dose of collagenase is administered. Exemplary treatment areas include, for example, the left buttock, the right buttock, or both the left and right buttocks.

[0040] The term "treatment session" is synonymous with "treatment visit" and includes a single visit to a physician's office at which collagenase or a formulation / composition containing collagenase is administered or a single self-administration period if the subject is self-administering collagenase or a formulation / composition containing collagenase. A treatment session or visit can include treating a single treatment area or treating multiple treatment areas. For example, in the case of the buttocks, a treatment session can include administering a specific total dose of collagenase to each of the left and right buttocks. Alternatively, a treatment session or visit can include administering a specific total dose of collagenase to only the left buttock.

[0041] The term "total dosage" refers to the total amount of collagenase administered to the treatment area in a given treatment session. For example, in the examples disclosed herein, the total dosage per treatment session refers to the total amount of collagenase administered to a single buttock per treatment session.

[0042] Method for reducing collagenase-mediated bruising in a subject having cellulite A method for reducing collagenase-mediated bruising in a subject having cellulite is disclosed herein. The method can include subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.42 mg per treatment session, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase can be administered using a three-injection technique (as described herein). In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Thus, the method can include subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.42 mg at a depth of about 0.5 inches using a three-injection technique per treatment session, thereby reducing collagenase-mediated bruising 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 session, and the treatment sessions can be separated by about three weeks. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions can be separated by about 42 days. The bruising can be reduced compared to the level of bruising associated with the administration of QWO® according to its U.S. prescribing information (www_accessdata.fda.gov / drugsatfda_docs / label / 2020 / 761146s000lbl.pdf; see also Example 2 herein). In some embodiments, the bruising can be reduced compared to the level of bruising associated with the subcutaneous administration of a total dose of collagenase of about 0.84 mg at a depth of 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject. In some embodiments, the bruising can be reduced compared to the level of bruising associated with the subcutaneous administration of a total dose of collagenase of about 0.84 mg at a depth of 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject, and the collagenase has a concentration of about 0.23 mg / ml.

[0043] A method for reducing collagenase-mediated bruising in a subject having cellulite comprises subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per 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 can be administered using a three-injection technique. In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Accordingly, a method for reducing collagenase-mediated bruising in a subject having cellulite comprises subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg at a depth of about 0.5 inches using a three-injection technique per 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 can receive more than one treatment session, and the treatment sessions are separated by about three weeks. The bruising can be reduced as compared to the level of bruising associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising can be reduced as compared to the level of bruising associated with subcutaneous administration of a total dose of collagenase of about 0.84 mg at a depth of about 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.23 mg / ml.

[0044] A method for reducing collagenase-mediated bruising in a subject having cellulite can include administering subcutaneously to a treatment area of the subject a total dose of collagenase of about 0.84 mg at a depth of about 0.25 inches per treatment session, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the collagenase can be administered using a three-injection technique. Thus, a method for reducing collagenase-mediated bruising in a subject having cellulite can include administering subcutaneously to a treatment area of the subject a total dose of collagenase of about 0.84 mg at a depth of about 0.25 inches per treatment session using a three-injection technique, thereby reducing collagenase-mediated bruising 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 session, and the treatment sessions can be separated by about three weeks. The bruising can be reduced as compared to the level of bruising associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising can be reduced as compared to the level of bruising associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase at a depth of about 0.5 inches per treatment session to the treatment area of the subject using a three-injection technique. In some embodiments, the bruising can be reduced as compared to the level of bruising associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase at a depth of about 0.5 inches per treatment session to the treatment area of the subject using a three-injection technique, and the collagenase has a concentration of about 0.23 mg / ml.

[0045] An exemplary three-injection technique is illustrated in FIG. 1. As shown in the figure, the subcutaneous injection can be administered as three 0.1 mL aliquots (total injection volume of 0.3 mL), with one aliquot administered perpendicular to the skin (A) and the other two aliquots administered at an angle of 45 degrees up (B) or down (C) with respect to the vertical axis. The depth of injection in FIG. 1 is 0.5 inches, although other injection depths, such as 0.25 inches, can also be used. Each syringe can contain 0.9 mL, allowing for three injections per syringe and up to 12 injections per treatment area.

[0046] The volume of each aliquot to be injected depends in part on the severity of the cellulite to be treated and the size of the treatment area. The total volume to be injected can be evenly divided among the respective aliquots. For example, if a pharmaceutical preparation of 0.3 ml is injected, each of the three aliquots can contain 0.1 ml. Suitable volumes for each aliquot include, 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.

[0047] A method for reducing collagenase-mediated bruising in a subject having cellulite may include subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing collagenase-mediated bruising 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. Thus, a method for reducing collagenase-mediated bruising in a subject having cellulite may include subcutaneously administering to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches using a single aliquot injection per treatment session, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, the administration may include injecting a single aliquot of collagenase up to 30 per treatment session into the treatment area. In some embodiments, the administration may include injecting a single aliquot of collagenase more than 30 per treatment session into the treatment area. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions may be separated by about 3 weeks. The bruising may be reduced compared to the level of bruising associated with the administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising may be reduced compared to the level of bruising associated with the subcutaneous administration of a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches using a single aliquot injection per treatment session into the treatment area of the subject, the collagenase having a concentration of about 0.23 mg / ml. In some embodiments, the administration may include injecting a single aliquot of collagenase up to 12 per treatment session into the treatment area. In some embodiments, the administration may include injecting a single aliquot of collagenase more than 12 per treatment session into the treatment area.

[0048] The number of injections administered via a single aliquot injection in a single treatment session is, in part, based on the severity of cellulite and the size of the treatment area. 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 a single treatment session.

[0049] When multiple injections are administered in a single treatment session, the injections may be spaced evenly or approximately evenly. The spacing of the injections depends, in part, on the volume being injected and the size of the treatment area. For example, the injections may be spaced 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 apart. In some embodiments, the injections are spaced about 2 cm to about 3 cm apart.

[0050] When multiple injections are administered in a single treatment session, the injections may be administered at random sites within the treatment area or at random sites and as one or more patterns within the treatment area. In some embodiments, the injections are spaced in a grid pattern. An exemplary grid pattern is shown in FIG. 3. As used herein, a grid pattern refers to a reproducible injection pattern and the injections are evenly spaced. The grid pattern can be, by way of example, hexagonal, circular, square, rectangular, triangular. In some embodiments, the injections in the grid pattern are spaced about 2 cm to about 3 cm apart. When injecting collagenase or a pharmaceutical formulation / composition containing collagenase at random sites, as one or more patterns, or as one or more patterns at random sites, the injection may contain a single aliquot or multiple aliquots of the pharmaceutical formulation for each site within the treatment area.

[0051] 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, each injection has a volume of about 0.1 ml to 0.3 ml.

[0052] Collagenase in any of the methods disclosed herein for reducing collagenase-mediated bruising can be a component of a composition / pharmaceutical formulation. In some embodiments, collagenase can be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose and tromethamine. The composition / pharmaceutical formulation may further contain HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation may have a pH of about 8.0.

[0053] A method for reducing collagenase-mediated bruising in a subject having cellulite may include subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, 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 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 subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, wherein, using a three-injection technique at a depth of about 0.5 inches per treatment session, a total dose of about 0.42 mg of collagenase is administered, thereby reducing collagenase-mediated bruising in the subject. The collagenase may have a concentration of about 0.23 mg / ml. The lidocaine may have a concentration of about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, or about 20 mg / mL. The epinephrine may have a concentration of about 5 mcg / mL or about 10 mcg / mL. In some embodiments, the composition may comprise about 0.23 mg / ml of collagenase, 2% lidocaine (20 mg / ml), and 1:200,000 epinephrine (5 mgc / ml). In some embodiments, the composition further comprises mannitol, sucrose, and tromethamine. Thus, the method may include subcutaneously administering to a treatment area of the subject a composition comprising 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, and the treatment sessions are separated by about three weeks. The bruising may be reduced compared to the level of bruising associated with administration of QWO® according to its U.S. prescribing information.In some embodiments, bruising can be reduced as compared to the level of bruising associated with subcutaneous administration of a composition comprising collagenase to a treatment area of a subject, and 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 can be reduced as compared to the level of bruising associated with subcutaneous administration of a composition comprising about 0.23 mg / ml of collagenase to a treatment area of a subject, and 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 is reduced as compared to the level of bruising associated with subcutaneous administration of a composition comprising collagenase, mannitol, sucrose, and tromethamine to a treatment area of a subject, and 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.

[0054] A method for reducing collagenase-mediated bruising in a subject having cellulite comprises subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.21 mg per 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 can be administered using a three-injection technique. In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Thus, a method for reducing collagenase-mediated bruising in a subject having cellulite comprises subcutaneously administering to a treatment area of the subject a total dose of about 0.21 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per 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 subject can receive more than one treatment session, and the treatment sessions are separated by about 42 days. The bruising can be reduced as compared to the level of bruising associated with the administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising can be reduced as compared to the level of bruising associated with the subcutaneous administration of a total dose of about 0.42 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session to the treatment area of the subject. In some embodiments, the bruising can be reduced as compared to the level of bruising associated with the subcutaneous administration of a total dose of about 0.42 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session to the treatment area of the subject, the collagenase having a concentration of about 0.23 mg / ml, and the subject having received more than one treatment session separated by about 42 days.

[0055] A method for reducing collagenase-mediated bruising in a subject having cellulite may include orally administering tranexamic acid to the subject per treatment session, and subcutaneously administering about 0.84 mg total dose of collagenase to the treatment area of the subject, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, tranexamic acid has a concentration of about 500 mg to about 1500 mg. In some embodiments, tranexamic acid has a concentration of about 1300 mg. Therefore, a method for reducing collagenase-mediated bruising in a subject having cellulite may include orally administering about 1300 mg of tranexamic acid to the subject per treatment session, and subcutaneously administering about 0.84 mg total dose of collagenase to the treatment area of the subject, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, collagenase may be administered using a three-injection technique. In some embodiments, collagenase may be administered at a depth of about 0.5 inches. A method for reducing collagenase-mediated bruising in a subject having cellulite may include orally administering about 1300 mg of tranexamic acid to the subject per treatment session, and subcutaneously administering about 0.84 mg total dose of collagenase at a depth of about 0.5 inches using a three-injection technique to the treatment area of the subject, thereby reducing collagenase-mediated bruising in the subject. In some embodiments, collagenase may have a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions are separated by about three weeks. In some embodiments, tranexamic acid is orally administered three times a day for five days during the first treatment session. In some aspects, during the first treatment session, tranexamic acid is orally administered to the subject three times on the day before the administration of collagenase, three times on the day of the administration of collagenase, and on each of the second, third, and fourth days after the administration of collagenase. In some embodiments, tranexamic acid is orally administered three times a day for five days during the second treatment session.In some scenarios, during the second treatment session, tranexamic acid is orally administered to the subject three times on the day prior to the administration of collagenase, three times on the day of the administration of collagenase, and on each of the 2nd, 3rd, and 4th days after the administration of collagenase. Bruising can be reduced as compared to the level of bruising associated with the administration of QWO® according to its U.S. prescribing information. In some embodiments, collagenase-mediated bruising is reduced as compared to the level of collagenase-mediated bruising associated with subcutaneous administration of a total dose of about 0.84 mg of collagenase at a depth of 0.5 inches using a three-injection technique per treatment session to the treatment area of the subject.

[0056] The level of bruising can be evaluated using the Investigator Assessment of Bruising Severity Scale (IABSS). An exemplary IABSS is illustrated in FIG. 2. As shown in FIG. 2, the exemplary IABSS includes five images showing bruising in the treatment areas (left and right buttocks) of one or more exemplary subjects, and the treatment area in each of the five images of the exemplary subject corresponds to the location of the treatment area of the subject to whom collagenase is administered, and each of the five images has a different bruising severity rating including numerical values and descriptions. The exemplary IABSS includes the following severity ratings: 0 - None or almost none: No bruising is observed or almost no bruising is observed; 1 - Mild: Mild bruising of pink to bright red color covering approximately 25% of the buttock; 2 - Moderate: Moderate bruising of dark red to purple color covering approximately 50% of the buttock; 3 - Severe: Severe bruising of dark purple color covering approximately 75% of the buttock; and 4 - Very severe: Severe bruising of dark purple to darkest purple color covering approximately 100% of the buttock.

[0057] Reduction in the level of bruising can include an improvement (i.e., decrease) of at least 1 point in the IABSS. Suitable reductions include, for example, a severity rating from 4 to 3, from 3 to 2, from 2 to 1, from 1 to 0, from 4 to 2, from 4 to 1, from 4 to 0, etc.

[0058] Method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject is disclosed herein. The method may include subcutaneously administering collagenase at a total dose of about 0.42 mg per treatment session to a treatment area of the subject, 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 subcutaneously administering collagenase at a total dose of about 0.42 mg per treatment session to a treatment area of the subject at a depth of about 0.5 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 have a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions may be separated by about three weeks. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions may be separated by about 42 days. The bruising and / or skin discoloration may be reduced as compared to the level of bruising and / or skin discoloration associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising and / or skin discoloration may be reduced as compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of collagenase at a total dose of about 0.84 mg at a depth of 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject. In some embodiments, the bruising and / or skin discoloration may be reduced as compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of collagenase at a total dose of about 0.84 mg at a depth of 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject, and the collagenase has a concentration of about 0.23 mg / ml.

[0059] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase can be administered using a three-injection technique. In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Thus, a method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions are separated by about three weeks. The bruising and / or skin discoloration can be reduced as compared to the level of bruising and / or skin discoloration associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising and / or skin discoloration can be reduced as 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 at a depth of about 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.23 mg / ml.

[0060] A method of reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering collagenase at a depth of about 0.25 inches per treatment session at a total dose of about 0.84 mg to a treatment area of the subject, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase can be administered using a three-injection technique. Thus, a method of reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering collagenase at a depth of about 0.25 inches per treatment session at a total dose of about 0.84 mg to a treatment area of the subject 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 can have a concentration of about 0.23 mg / ml. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions are separated by about three weeks. The bruising and / or skin discoloration can be reduced as compared to the level of bruising and / or skin discoloration associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising and / or skin discoloration can be reduced as 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 at a depth of about 0.5 inches per treatment session using a three-injection technique to a treatment area of the subject. In some embodiments, the bruising and / or skin discoloration can be reduced as 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 at a depth of about 0.5 inches per treatment session using a three-injection technique to a treatment area of the subject, and the collagenase has a concentration of about 0.23 mg / ml.

[0061] The volume of each aliquot to be injected depends, in part, on the severity of the cellulite to be treated and the size of the treatment area. The total volume to be injected can be evenly divided among the respective aliquots. For example, if 0.3 ml of a pharmaceutical preparation is injected, each of the three aliquots can contain 0.1 ml. Suitable volumes for each aliquot include, 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.

[0062] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, wherein the collagenase has a concentration of about 0.09 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of 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. Thus, a method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches using a single aliquot injection per treatment session, wherein the collagenase has a concentration of about 0.09 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the administration comprises from 1 to 30 single aliquot injections of collagenase into the treatment area per treatment session. In some embodiments, the administration comprises more than 30 single aliquot injections of collagenase into the treatment area per treatment session. In some embodiments, the subject can receive more than 1 treatment session, and the treatment sessions are separated by about 3 weeks. The bruising and / or skin discoloration can be reduced compared to the level of bruising and / or skin discoloration associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the 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 at a depth of about 0.25 inches using a single aliquot injection per treatment session to a treatment area of the subject, wherein the collagenase has a concentration of about 0.23 mg / ml. In some embodiments, the administration comprises from 1 to 12 single aliquot injections of collagenase into the treatment area per treatment session. In some embodiments, the administration comprises more than 12 single aliquot injections of collagenase into the treatment area per treatment session.

[0063] The number of injections administered by injection of a single aliquot in a single treatment session is based in part on the severity of cellulite and the size of the treatment area. 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 than 100 injections can be administered to the treatment area in a single treatment session.

[0064] When multiple injections are administered in a single treatment session, the injections can be spaced evenly or approximately evenly. The spacing of the injections depends in part on the injection volume and the size of the treatment area. For example, the injections can be separated by 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 injections are spaced about 2 cm to about 3 cm apart.

[0065] 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 patterns. In some embodiments, the injections are spaced in a grid pattern. In some embodiments, the injections in the grid pattern are spaced about 2 cm to about 3 cm apart. When injecting collagenase or a pharmaceutical formulation / composition containing collagenase at random sites, as one or more patterns or at random sites and as one or more patterns, the injection can include a single aliquot or multiple aliquots of the pharmaceutical formulation to each site within the treatment area.

[0066] Suitable volumes for each injection in 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, each injection has a volume of about 0.1 ml to 0.3 ml.

[0067] Collagenase in any of the methods disclosed herein for reducing bruising and / or skin discoloration can be a component of a composition / pharmaceutical formulation. In some embodiments, collagenase can be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose and tromethamine. The composition / pharmaceutical formulation may further comprise HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation can have a pH of about 8.0.

[0068] A method of reducing bruising and / or skin discoloration associated with cellulite collagenase-mediated treatment in a subject may include subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, wherein a total dose of about 0.42 mg of collagenase is administered per treatment session, thereby reducing bruising and / or skin discoloration associated with cellulite collagenase-mediated treatment 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 of reducing bruising and / or skin discoloration associated with cellulite collagenase-mediated treatment in a subject may include subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, 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 per treatment session, thereby reducing bruising and / or skin discoloration associated with cellulite collagenase-mediated treatment in the subject. The collagenase may have a concentration of about 0.23 mg / ml. The lidocaine may have a concentration of about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, or about 20 mg / mL. The epinephrine may have a concentration of about 5 mcg / mL or about 10 mcg / mL. In some embodiments, the composition may comprise about 0.23 mg / ml of collagenase, 2% lidocaine (20 mg / ml), and 1:200,000 epinephrine (5 mgc / ml). In some embodiments, the composition further comprises mannitol, sucrose, and tromethamine. Thus, the method may include subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, epinephrine, mannitol, sucrose, and tromethamine, 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 per treatment session, thereby reducing bruising and / or skin discoloration associated with cellulite collagenase-mediated treatment in the subject. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions are separated by about three weeks.Contusions and / or skin discoloration may be reduced compared to the level of contusions and / or skin discoloration associated with administration of QWO® according to its US prescribing information. In some embodiments, contusions and / or skin discoloration may be reduced compared to the level of contusions and / or skin discoloration associated with subcutaneous administration of a composition comprising collagenase to the treatment area of a subject, and 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, contusions and / or skin discoloration may be reduced compared to the level of contusions and / or skin discoloration associated with subcutaneous administration of a composition comprising about 0.23 mg / ml of collagenase to the treatment area of a subject, and 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, contusions and / or skin discoloration are reduced compared to the level of contusions and / or skin discoloration associated with subcutaneous administration of a composition comprising collagenase, mannitol, sucrose and tromethamine to the treatment area of a subject, and 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.

[0069] A method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.21 mg per treatment session, the collagenase having a concentration of about 0.12 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the collagenase can be administered using a three-injection technique. In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Thus, a method for reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject comprises subcutaneously administering to a treatment area of the subject a total dose of about 0.21 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session, the collagenase having a concentration of about 0.12 mg / ml, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions are separated by about 42 days. The bruising and / or skin discoloration can be reduced as compared to the level of bruising and / or skin discoloration associated with administration of QWO® according to its U.S. prescribing information. In some embodiments, the bruising and / or skin discoloration can be reduced as compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a total dose of about 0.42 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject. In some embodiments, the bruising and / or skin discoloration can be reduced as compared to the level of bruising and / or skin discoloration associated with subcutaneous administration of a total dose of about 0.42 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.23 mg / ml, and the subject having received more than one treatment session separated by about 42 days.

[0070] A method of reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in a subject may include orally administering tranexamic acid to the subject at a dose of about 1300 mg per treatment session, and subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of the subject, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, tranexamic acid has a concentration of about 500 mg to about 1500 mg. In some embodiments, tranexamic acid has a concentration of about 1300 mg. Thus, a method of reducing bruising 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 the subject per treatment session, and subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of the subject, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite 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. A method of reducing bruising 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 the subject, and subcutaneously administering a total dose of about 0.84 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session to the treatment area of the subject, thereby reducing bruising and / or skin discoloration associated with collagenase-mediated treatment of cellulite in the subject. In some embodiments, collagenase can have a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions are separated by about three weeks. In some embodiments, tranexamic acid is orally administered three times a day for five days during the first treatment session. In some aspects, during the first treatment session, tranexamic acid is orally administered to the subject three times on the day before collagenase administration, three times on the day of collagenase administration, and on the second, third, and fourth days after collagenase administration.In some embodiments, tranexamic acid is orally administered three times a day for 5 days during the second treatment session. In some aspects, during the second treatment session, tranexamic acid is orally administered to the subject three times on the day prior to the administration of collagenase, three times on the day of the administration of collagenase, and on each of the 2nd, 3rd, and 4th days after the administration of collagenase. Bruising and / or skin discoloration may be reduced compared to the level of bruising and / or skin discoloration associated with the administration of QWO® according to its U.S. prescribing information. In some embodiments, bruising and / or skin discoloration is reduced compared to the level of bruising and / or skin discoloration associated with the subcutaneous administration of collagenase at a total dose of about 0.84 mg at a depth of 0.5 inches using a three-injection technique per treatment session to the treatment area of the subject.

[0071] Reduction in the level of bruising and / or skin discoloration may include an improvement (i.e., decrease) of at least 1 point in the IABSS. Suitable reductions include, for example, a severity rating of 4 to 3, 3 to 2, 2 to 1, 1 to 0, 4 to 2, 4 to 1, 4 to 0, etc.

[0072] Treatment method for cellulite A method of treating cellulite in a subject is disclosed herein. The method of treating cellulite in a subject can include the step of subcutaneously administering collagenase at a total dose of about 0.42 mg per treatment session to a treatment area of the subject, thereby treating cellulite in the subject. In some embodiments, the collagenase can be administered using a three - injection technique. In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Thus, the method of treating cellulite in a subject can include the step of subcutaneously administering collagenase at a total dose of about 0.42 mg per treatment session at a depth of about 0.5 inches using a three - injection technique to a treatment area of the subject, thereby treating cellulite in the subject. In some embodiments, the collagenase has a concentration of about 0.23 mg / ml. In some embodiments, the collagenase can be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation can further include HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation can have a pH of about 8.0. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions are separated by about three weeks. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions are separated by about 42 days.

[0073] A method of treating cellulite in a subject may include administering subcutaneously to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.05 mg / ml, 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 of treating cellulite in a subject may include administering subcutaneously to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session, the collagenase having a concentration of about 0.05 mg / ml, thereby treating cellulite in the subject. In some embodiments, the collagenase may be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation may further include HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation may have a pH of about 8.0. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions may be separated by about three weeks.

[0074] A method of treating cellulite in a subject may include administering subcutaneously to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches per treatment session, the collagenase having a concentration of about 0.23 mg / ml, thereby treating cellulite in the subject. In some embodiments, the collagenase may be administered using a three-injection technique. Thus, a method of treating cellulite in a subject may include administering subcutaneously to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches per treatment session using a three-injection technique, the collagenase having a concentration of about 0.23 mg / ml, thereby treating cellulite in the subject. In some embodiments, the collagenase may be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation may further include HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation may have a pH of about 8.0. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions may be separated by about three weeks.

[0075] A method of treating cellulite in a subject can include administering subcutaneously to a treatment area of the subject a total dose of collagenase of about 0.84 mg per treatment session, the collagenase having a concentration of about 0.09 mg / ml, thereby treating 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. Thus, a method of treating cellulite in a subject can include administering subcutaneously to a treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches using a single aliquot injection per treatment session, the collagenase having a concentration of about 0.09 mg / ml, thereby treating cellulite in the subject. In some embodiments, the collagenase can be a component of a composition / pharmaceutical formulation comprising collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation can further include HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation can have a pH of about 8.0. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions can be separated by about 3 weeks.

[0076] The number of injections administered by a single aliquot injection in a single treatment session is based in part on the severity of the cellulite and the size of the treatment area. 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 than 100 injections can be administered to the treatment area in a single treatment session. In some embodiments, the administration includes up to 30 single aliquot injections of collagenase to the treatment area per treatment session.

[0077] When multiple injections are administered in a single treatment session, the injections can be spaced evenly or approximately evenly apart. The spacing between the injections depends in part on the volume being injected and the size of the treatment area. For example, the injections can be spaced apart by 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 injections are spaced about 2 cm to about 3 cm apart.

[0078] 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 patterns. In some embodiments, the injections are spaced in a grid pattern as described herein.

[0079] Suitable volumes for each injection in 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, each injection has a volume of about 0.1 ml to 0.3 ml.

[0080] A method for treating cellulite in a subject may include the step of subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, wherein a total dose of collagenase of about 0.42 mg is administered per treatment session, 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 the step of subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, 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 per treatment session, thereby treating cellulite in the subject. In some embodiments, the composition comprises 0.23 mg / ml of collagenase, 2% lidocaine, and 1:200,000 epinephrine. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions are separated by about three weeks. In some embodiments, the composition further comprises mannitol, sucrose, and tromethamine. Thus, a method for treating cellulite in a subject may include the step of subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, epinephrine, mannitol, sucrose, and tromethamine, 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 per treatment session, thereby treating cellulite in the subject.

[0081] A method of treating cellulite in a subject can include the step of subcutaneously administering collagenase at a total dosage of about 0.21 mg per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.12 mg / ml, thereby treating cellulite in the subject. In some embodiments, the collagenase can be administered using a three-injection technique. In some embodiments, the collagenase can be administered at a depth of about 0.5 inches. Thus, a method of treating cellulite in a subject can include the step of subcutaneously administering collagenase at a total dosage of about 0.21 mg at a depth of about 0.5 inches using a three-injection technique per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.12 mg / ml, thereby treating cellulite in the subject. In some embodiments, the subject can receive more than one treatment session, and the treatment sessions are separated by about 42 days. In some embodiments, the collagenase can be a component of a composition / pharmaceutical formulation that includes collagenase, mannitol, sucrose, and tromethamine. The composition / pharmaceutical formulation can further include HCl for adjusting the pH if necessary. The composition / pharmaceutical formulation can have a pH of about 8.0.

[0082] A method for treating 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 treatment area of the subject per treatment session, thereby treating cellulite in the subject. In some embodiments, tranexamic acid has a concentration of about 500 mg to about 1500 mg. In some embodiments, tranexamic acid has a concentration of about 1300 mg. Thus, a method for treating cellulite in a subject may include orally administering about 1300 mg of tranexamic acid to the subject per treatment session and subcutaneously administering a total dose of about 0.84 mg of collagenase to the treatment area of the subject, thereby treating cellulite in the subject. In some embodiments, collagenase may be administered using a three - injection technique. In some embodiments, collagenase may be administered at a depth of about 0.5 inches. A method for treating cellulite in a subject may include orally administering about 1300 mg of tranexamic acid to the subject per treatment session and subcutaneously administering a total dose of about 0.84 mg of collagenase at a depth of about 0.5 inches using a three - injection technique to the treatment area of the subject, thereby treating cellulite in the subject. In some embodiments, collagenase may have a concentration of about 0.23 mg / ml. In some embodiments, the subject may receive more than one treatment session, and the treatment sessions are separated by about three weeks. In some embodiments, tranexamic acid is orally administered three times a day for five days during the first treatment session. In some aspects, during the first treatment session, tranexamic acid is orally administered to the subject three times on the day before the administration of collagenase, three times on the day of the administration of collagenase, and on each of the second, third, and fourth days after the administration of collagenase. In some embodiments, tranexamic acid is orally administered three times a day for five days during the second treatment session. In some aspects, during the second treatment session, tranexamic acid is orally administered to the subject three times on the day before the administration of collagenase, three times on the day of the administration of collagenase, and on each of the second, third, and fourth days after the administration of collagenase.

[0083] The treatment of cellulite can be established by scales or measurement tools selected from the Hexsel Cellulite Severity Scale (Hexsel CSS), Hexsel Depression Depth Score, Recart Scale, Dimple Analysis, Clinician-Reported Photographic Numerical Cellulite Severity Scale (CR-PCSS), Patient-Reported Photographic Numerical 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), Abbreviated PR-CIS (PR-CIS Abbreviated), Subject Self-Rating Scale (SSRS), Subject Satisfaction with Cellulite Treatment (SSCT), Clinician Evaluation of Cellulite Severity (photographs or other images), Body-Q, and validated photographic numerical or other scales used by clinicians and / or patients to evaluate cellulite severity, improvement, and / or patient satisfaction.

[0084] The Fitzpatrick skin scale is a 6-level scale (levels I - VI) for the evaluation of skin color and tendency to sunburn for classifying skin types. Skin types range from level I: thin white skin, blue / light brown eyes, blond / red hair, always burns, never tans, to level VI: dark brown or black skin, does not burn, always tans darkly. The investigator (or designated human) determines the Fitzpatrick skin type for all subjects at the time of screening.

[0085] The Hexsel Cellulite Severity Scale (CSS) is a photographic numerical scale used to evaluate five major morphological cellulite characteristics: (A) number of visible depressions, (B) depth of depressions, (C) morphological appearance of skin surface changes, (D) skin laxity, looseness or sagging, and (E) current classification scale based on medical literature (Hexsel et al., 2009; Nuernberger and Mueller, 1978). Each of these characteristics is evaluated on a 4-point scale from low of 0 to high of 3 (shown in Table 1).

Table 1

[0086] The Clinician-Reported Photographic Numerical Cellulite Severity Scale (CR-PCSS) is a validated 5-level photographic numerical scale specifically developed for and used by investigators to assess the severity of cellulite in the treatment area (e.g., one or both buttocks) of participants by live evaluation. The ratings range from 0 (none) to 4 (severe) with labels and descriptors to assist the investigator in the evaluation. This evaluation should be made with the participant standing in a relaxed gluteal muscle position.

[0087] The Patient-Reported Photographic Numerical Cellulite Severity Scale (PR-PCSS) is a validated 5-level photographic numerical scale specifically developed for and used by patients to assess the severity of the patient's cellulite in the treatment area (e.g., one or both buttocks) by viewing each digital image of the patient's buttocks captured by photography. The ratings range from 0 (none) to 4 (severe) with images, labels and descriptors to assist the patient in the evaluation.

[0088] The Investigator Global Aesthetic Improvement Scale (I-GAIS) is a 7-level scale ranging from 3 (very large improvement) to -3 (very large worsening) (Table 2). Investigator physicians use the I-GAIS to determine the degree of improvement in each treatment area by comparing cellulite from the pre-treatment (baseline) image on day 1 of treatment to the images taken at subsequent visits for each treatment area. The I-GAIS evaluation is based on digital photographs and is performed separately for each of the two treatment areas. [Table 2]

[0089] Cellulite Evaluation of Subjects and Investigators - The cellulite evaluation of the investigator is independent of the evaluation of the subject. Therefore, the cellulite evaluation of all subjects must be completed before the cellulite evaluation of the investigator is initiated. The subject evaluation is made with the subject alone and in the absence of test site personnel in the room. The investigator is instructed not to verbalize the investigator rating in the presence of the subject and vice versa.

[0090] The Subject Global Aesthetic Improvement Scale (S-GAIS) is a 7-level scale ranging from 3 (very large improvement) to -3 (very large deterioration). The subject uses the S-GAIS to determine the degree of improvement in each treatment area by comparing cellulite from the pre-treatment (baseline) image on day 1 of each treatment area to the images taken at subsequent visits.

[0091] The Body-Q assessment of cellulite is a subset of questions from the Body-Q questionnaire developed to measure patient understanding of weight loss and / or creation of body contours.

[0092] Investigator satisfaction using the Administration scale (Likert scale) is a 5-point scale that can be used to determine investigator satisfaction by CCH (Investigator Satisfaction by CCH Administration).

[0093] Ultrasound Evaluation - Surface ultrasound images of the treatment area can be captured. Predetermined parameters can be measured / evaluated from the images.

[0094] 3-D Photography - The PRIMOS 3-D camera can be utilized as an investigational tool to objectively quantify skin surface roughness scores. This equipment and related imaging technology present a novel method for capturing changes in skin relaxation and the dissipation of skin depressions due to cellulite.

[0095] Digital photography can be performed using a supplied canfield camera system with the subject standing upright in a consistent, standard, relaxed posture and with the gluteal muscles relaxed.

[0096] Treatment of cellulite can include at least a 1-point improvement in any of the above metrics. Using CR-PCSS as an example, treatment of cellulite can include a severity rating change from 4 to 3, from 3 to 2, from 2 to 1, from 1 to 0, from 4 to 2, from 4 to 1, from 4 to 0, etc.

[0097] In any of the methods disclosed herein, a subject can receive a plurality of treatment sessions. For example, a subject can receive 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more treatment sessions. In some embodiments, treatment sessions are separated by 3 weeks (21 days). In some embodiments, treatment sessions are separated by 6 weeks (42 days).

[0098] The amount of collagenase injected using the three - injection technique refers to the amount of collagenase in each of the combined three injection aliquots. The amount of collagenase injected using the single - aliquot technique refers to the amount of collagenase in the single aliquot. 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., three - injection technique or single - aliquot injection), and the volume administered per injection. Suitable amounts of collagenase administered to the treatment area per injection for any of the methods disclosed herein 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.

[0099] The amount of collagenase administered in a single treatment session depends in part on the size of the treatment area, the severity of cellulite, the injection technique, the volume administered per injection, and the expected number of treatment sessions. Suitable amounts of collagenase to be administered to the treatment area in a single treatment session for any of the disclosed methods 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 0.84 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 than 20.0 mg. 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 administered to the treatment area in a single treatment session. In some embodiments, about 0.42 mg of collagenase is administered to the treatment area in a single treatment session. In some embodiments, about 0.84 mg of collagenase is administered to the treatment area in a single treatment session.

[0100] "Collagenase" means any of the following: (a) collagenase having the activity defined in EC 3.4.24.3 (www_brenda-enzymes.org / enzyme.php?ecno=3.4.24.3 (accessed on April 27, 2023)) (including variants); (b) collagenase produced by fermentation of Clostridium histolyticum (also known as Hathewaya histolytica); (c) CCH (described herein); (d) collagenase having at least 50% sequence alignment with collagenase I (also referred to as class I collagenase) when determined by BLAST; (e) collagenase having at least 50% sequence alignment with collagenase II (also referred to as class II collagenase) when determined by BLAST; (f) collagenase produced by fermentation of other supply organisms (i.e., non-Clostridium histolyticum), such as mammalian, crustacean, fungal, bacterial or microbial collagenase; (g) collagenase obtained by recombinant technology; (h) collagenase having a molecular mass of about 65 kDa to about 130 kDa; (i) collagenase designated as collagenase I (col I) or collagenase II (col II); (j) a mixture of collagenase I and II; (k) collagenase or its derivative derived from strain JCM 1403 (ATCC 19401); (l) collagenase or its derivative derived from strain ATCC 21000; (m) collagenase or its derivative derived from ATCC 69334; (n) collagenase derived from Clostridium perfringens; (o) collagenase derived from Vibrio alginolyticus; (p) collagenase derived from the genus Streptomyces; (q) collagenase derived from the genus Pseudomonas; (r) collagenase derived from Achromobacter iophagus; (s) collagenase described in Worthington Biochemical Corp. (www_Worthington-biochem; "Product Highlights"); (t) collagenase described in Sigma-Aldrich (www_sigma-aldrich); (u) the following characteristics: ● Approximately 0.08 - 7.70 (SRC assay; described in International Publication No. WO2020 / 058755) or approximately 0.3 - 30.5 (GPA assay; described in International Publication No. WO2020 / 058755) of V max (min -1 ); ● Approximately 4.1 - 410 nM (SRC assay) or approximately 0.03 - 3.1 mM (GPA assay) of K M ; ● Approximately 1.1 - 107 (SRC assay) or approximately 93 - 9,179 (GPA assay) of K cat (sec -1 ); ● Approximately 376 - 37,222 (SRC assay) or approximately 4 - 428 (GPA assay) of 1 / K cat (microsecond); or ● Approximately 5,140 - 508,814 (SRC assay) or approximately 60 - 5,934 (GPA assay) of K cat / K M (mM -1 sec -1 ) and having one or more of a collagenase; (v) Collagenase described in Nordmark Arzneimittel GmbH & Co. KG; (w) Collagenase derived from Stock 004; (x) Biosimilar of the collagenase component of QWO® or biosimilar of QWO®; (y) Biosimilar of the collagenase component of XIAFLEX® or biosimilar of XIAFLEX®; or (z) Any equivalent or mixture of the foregoing. Non-limiting examples of collagenases that may be used in the disclosure herein are described in U.S. Patent No. 7,811,560, U.S. Patent No. 9,757,435, U.S. Patent No. 9,744,138, and International Publication No. WO2012 / 125948.

[0101] In some embodiments, the collagenase may include collagenase I. Suitable collagenase I includes, for example, a collagenase I comprising 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, the collagenase I includes the amino acid sequence of SEQ ID NO:1.

[0102] In some embodiments, the collagenase may include collagenase II. Suitable collagenase II includes, for example, a collagenase II comprising 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:2. In some aspects, the collagenase II includes the amino acid sequence of SEQ ID NO:2. [Table 3]

[0103] In some embodiments, the collagenase can include a mixture of collagenase I and collagenase II. The collagenase can include collagenase I having an amino acid sequence with 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO:1 and collagenase II having an amino acid sequence with 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SEQ ID NO:2. In some aspects, the collagenase includes a mixture of collagenase I having the amino acid sequence of SEQ ID NO:1 and collagenase II having the amino acid sequence of SEQ ID NO:2. Suitable mixtures of collagenase I and collagenase II can 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 can have a purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% when measured, for example, by reverse phase HPLC.

[0104] In some embodiments, the collagenase can include collagenase Clostridium histolyticum (CCH). As used herein, "CCH" refers to collagenase Clostridium histolyticum that includes a mixture of collagenase I (SEQ ID NO:1) and collagenase II (SEQ ID NO:2) at a mass ratio of about 1:1. CCH is obtained by fermentation of Clostridium histolyticum (also known as Hathewaya histolytica).

[0105] In some embodiments, collagenase I and collagenase II have the following characteristics: Collagenase I (SRC microplate assay) 〇V max: Approximately 0.08 - 7.70 min -1 OK M : Approximately 4.1 - 410 nM OK cat : Approximately 1.1 - 107 sec -1 〇1 / K cat : Approximately 376 - 37,222 microseconds OK cat / K M : Approximately 5,140 - 508,814 mM -1 sec -1 Collagenase II (GPA microplate assay) 〇V max : Approximately 0.3 - 30.5 min -1 OK M : Approximately 0.03 - 3.1 mM OK cat : Approximately 93 - 9,179 sec -1 〇1 / K cat : Approximately 4 - 428 microseconds OK cat / K M : Approximately 60 - 5,934 mM -1 sec -1 may have.

[0106] In some embodiments, Collagenase I and Collagenase II have the following characteristics: Collagenase I (SRC assay): 〇V max : Approximately 3.8 min -1 OK M : Approximately 2.07x10 -4 mM OK cat : Approximately 53 sec -1 〇1 / K cat : Approximately 18,799 microseconds 〇k cat / K M : Approximately 256,977 mM -1 sec -1 Collagenase II (GPA assay): 〇V max : about 15.4 min -1 〇K M : about 1.6 mM 〇K cat : about 4,636 sec -1 〇1 / K cat : about 216 microseconds 〇k cat / K M : about 2,997 mM -1 sec -1 may have.

[0107] Collagenase-containing preparation Preparations containing collagenase; lidocaine and epinephrine are disclosed herein.

[0108] Any of the above-mentioned collagenases may be used in the preparation. In some embodiments, the collagenase may include collagenase I. In some embodiments, the collagenase may include collagenase II. In some embodiments, the collagenase may include a mixture of collagenase I and collagenase II. In some aspects, the collagenase includes a mixture of collagenase I having the amino acid sequence of SEQ ID NO: 1 and collagenase II having the amino acid sequence of SEQ ID NO: 2. In some embodiments, the collagenase may include collagenase Clostridium histolyticum (CCH).

[0109] The collagenase may have a concentration of about 0.23 mg / ml. Lidocaine may have a concentration of about 5 mg / mL, about 10 mg / mL, about 15 mg / mL or about 20 mg / mL. Epinephrine may have a concentration of about 5 mcg / mL or about 10 mcg / mL. In some embodiments, the preparation may include about 0.23 mg / ml of collagenase, about 2% lidocaine and about 1:200,000 epinephrine.

[0110] The formulation may further contain mannitol, sucrose and tromethamine. Thus, in some embodiments, the formulation comprises collagenase, lidocaine, epinephrine, mannitol, sucrose and tromethamine.

Example

[0111] Example The following examples are provided to further illustrate some of the aspects disclosed herein. The examples are intended to illustrate, without limitation, the disclosed aspects.

[0112] Example 1 - APHRODITE-1 Trial Background Collagenase Clostridium histolyticum-aaes (CCH-aaes; QWO®, Endo Aesthetics LLC) for injection is indicated for the treatment of moderate to severe cellulite in the buttocks of adult women. CCH-aaes injection is thought to initiate Enzymatic Subcision and Remodeling® which involves the lysis of mature collagen-rich septa that cause the characteristic dimpling of cellulite, rapid onset of neocollagenesis and some degree of adipose lobule reorganization (into smaller and more uniform-sized lobules). Injection site bruising was the most common adverse event reported in a pooled analysis of two identically designed, randomized Phase 3 trials in women with cellulite in the buttocks treated with CCH-aaes (84%) or placebo (21%). During these trials, 0.84 mg of CCH-aaes was administered per treatment area using a 1 / 2-inch 3-aliquot technique (also referred to as the 3-injection technique illustrated in Figure 1). Injection site bruising after CCH-aaes administration generally resolves within 14 to 21 days, but the bruising is bothersome to some women and may be associated with subsequent skin discoloration. Therefore, a study was designed to identify a potent intervention that could reduce bruising and its associated consequences.

[0113] Objective To evaluate the effects of several interventions on the prevention or reduction of bruising in the buttocks of women with moderate to severe cellulite after treatment with CCH-aaes injection, using a self-controlled trial design.

[0114] Methods Trial design and interventions APHRODITE-1 (a phase 2 open-label self-controlled trial of different interventions to reduce bruising after CCH-aaes treatment) planned to enroll up to 150 women during this first stage of the trial. The trial was designed to evaluate different CCH-aaes doses, different CCH-aaes concentrations, different injection techniques (e.g., injection depth), and diluent additives in efforts to prevent or reduce injection site bruising. At later stages of the trial, if desired, variables such as the concentration of other diluent additives, alternative doses, and injection timing, e.g.: - Different CCH-aaes doses or different concentrations; - Different injection techniques (e.g., injection depth); - Inclusion of diluent additives; and / or - Addition of a new intervention cohort were considered, and the trial was designed to allow for the addition of new intervention cohorts.

[0115] Up to 30 participants were assigned to each of 8 cohorts (in a 1:1:1:1:1:1 ratio) using an interactive response technology system shown in Table 4:

Table 4-1

Table 4-2

Table 4-3

[0116] The patient population was: - Up to 180 women aged 18 to 60 years; - 18 to <32 kg / m 2 body mass index (BMI); - Moderate to severe cellulite on both hips (CR-PCSS rating of 3 or 4); - Hexsel CSS total score of ≦12; and - Fitzpatrick skin types I to IV were included.

[0117] Table 5 provides a summary of the demographics and baseline characteristics for these subjects enrolled in the study.

Table 5-1

Table 5-2

Table 5-3

Table 5-4

[0118] Throughout the study, injection site bruising (investigational and control sites) as well as other safety and efficacy parameters were monitored. The study included four follow-up visits after each CCH-aaes treatment session (1 to 2 days, 3 to 5 days, 6 to 9 days, and 10 to 14 days post-treatment) to assess injection site bruising severity and document severity using standardized digital photographs (Figures 4A and 4B).

[0119] Ongoing (ad hoc) assessments for specific issues At the time of this application, registration for the trial was complete, but all follow-up visits had not been conducted. An assessment was made for specific issues regarding the data generated from the follow-up visits completed up to that date. The severity of bruising was determined by live evaluation using the 5-point Investigator Assessment of Bruise Severity Scale (IABSS), and each buttock was evaluated individually to enable comparison (Figure 2). The first efficacy endpoint was the proportion of participants in which the left buttock (investigational treatment) had an IABSS score that was at least 1 level lower than the right buttock (control treatment) 3 to 5 days after the first CCH-aaes treatment (second follow-up visit) of the post-injection series 1. The second efficacy endpoint was: - The proportion of participants in which the left buttock of the participant had an IABSS score that was at least 1 level lower than the right buttock at the visit; - The proportion of participants who had an improvement of ≥1 level from baseline (day 1) in the Investigator Global Aesthetic Improvement Scale (I-GAIS) at the visit (I-GAIS uses a 7-point scale ranging from +3 (“very much improved”) to -3 (“very much worsened”)); including.

[0120] Adverse events, including injection site reactions, were monitored at each buttock site throughout the trial.

[0121] Conclusion APHRODITE-1 was designed to determine whether different interventions (i.e., dose, concentration, injection technique, and additional diluent additives) can prevent or reduce injection site bruising after CCH-aaes treatment for cellulite in the buttocks. This trial was created with flexibility to add cohorts, so additional interventions can be tested over time.

[0122] The data provided below is an extraction for specific issues of the data collected up to the time of this application (before completion of all follow-up visits). Table 6 shows a summary of level 1 IABSS responders at visit 2 (days 1 - 2 after treatment), visit 3 (days 3 - 5 after treatment), and visit 4 (days 6 - 9 after treatment) for cohorts 1 - 6. For cohorts 7a and 7b, comparisons of equivalent visits for each buttock are listed (visit 2 and visit 7; visit 3 and visit 8; visit 4 and visit 9). The data was collected up to 90 days ± 7 days, but the level of post-treatment bruising began to be investigated for both the study side (left buttock) and the control side (right buttock around visit 4, making any visual differences in bruising between the two buttocks negligible. More precisely, bruising began around visit 4 for cohorts 1 - 6 and around visit 4 / 9 for cohorts 7a and 7b (the corresponding post-treatment visit for each buttock). Therefore, only data from visits 2, 3, and 4 (for cohorts 1 - 6) and visits 2 vs 7 / 7 vs 2, 3 vs 8 / 8 vs 3, and 4 vs 9 / 9 vs 4 (for cohorts 7a and 7b) are provided in this specification. As shown in Table 6, at visit 3, over 90% of the subjects in cohort 1 showed at least a 1-point improvement in IABSS, and over 63% of the 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 for visit 3, so bruising was drawn for both the study side and the control side at visit 4 as described above.

[0123] Figure 5 shows the level of bruising observed in a patient from cohort 1 at visit 3 (days 3 - 5 after treatment). The left buttock received 0.42 mg of CCH-aaes at a concentration of 0.23 mg / ml using a three-injection technique, and the right buttock (control) received 0.84 mg of CCH-aaes at a concentration of 0.23 mg / ml using a three-injection technique. As shown in Figure 5, at least a 1-point improvement was observed in IABSS.

Table 6-1

Table 6-2

[0124] Example 2 - QWO (Registered Trademark) US Prescription Information (QWO (Registered Trademark) Label) A copy of the FDA-approved label for QWO is provided below, which provides US prescription information. This label was obtained from www_accessdata.fda.gov / drugsatfda_docs / label / 2020 / 761146s000lbl.pdf on May 10, 2023.

[0125] Highlights of Prescription Information These highlights do not include all the information necessary to use QWO safely and effectively. Refer to the full prescribing information for QWO. QWO for Injection, for Subcutaneous Use TM (Collagenase Clostridium histolyticum-aaes) First US Approval: 2020 Indications and Usage QWO is a combination of bacterial collagenases indicated for the treatment of moderate to severe cellulite in the buttocks of adult women. (1) Dosage and Administration ● The treatment area is defined as a single buttock receiving up to 12 injections of up to 0.3 mL each of QWO (up to a total of 3.6 mL). (2.1) ● Treatment visits can consist of up to 2 treatment areas. Treatment visits should be repeated every 21 days between 3 treatment visits. (2.1) ● Reconstitute the QWO lyophilized powder with the supplied diluent before use. (2.2) ● Inject 0.84 mg of QWO per treatment area as 12 subcutaneous injections (administered as 0.3 mL injections given as 3 x 0.1 mL aliquots per injection). (2.3) Dosage Form and Strength For injection: Lyophilized powder in single-dose vials of 0.92 mg or 1.84 mg. (3) Contraindications ● History of hypersensitivity to any collagenase or any of the components in the formulation. (4) ● Infection at the injection site. (4) Warnings and Precautions ● Hypersensitivity Reactions: Severe hypersensitivity reactions, such as anaphylaxis, can occur with collagenase clostridium histolyticum. If a severe hypersensitivity reaction occurs, initiate appropriate treatment. (5.1) ● Injection Site Bruising: Bruising occurs frequently after QWO administration. Use with caution in patients with bleeding disorders or who are currently being treated with antiplatelet agents (except those receiving ≤150 mg of aspirin daily) or anticoagulant therapy. (5.2) ● Substitution: QWO should not be substituted for other injectable collagenase products. QWO is not indicated for the treatment of Peyronie's disease or Dupuytren's contracture. (5.3) Adverse Reactions The most common adverse reactions (≥1%) were injection site related (bruising, pain, nodules, pruritus, erythema, rash, swelling, and excitation). (6.1) To report a suspected adverse reaction, contact Endo Pharmaceuticals Inc. at 1-800-462-3636 or the FDA at 1-800-FDA-1088 or 1732259873033_0 contact the FDA at 1732259873033. See 17 for Patient Counseling Information and FDA-approved Patient Labeling Revision: 07 / 2020 Sufficient Prescription Information: Contents * 1 Indications and Usage 2 Dosage and Administration 2.1 Dosage 2.2 Reconstitution of Lyophilized Powder 2.3 Administration 3 Dosage Form and Strength 4 Contraindications 5 Warnings and Precautions 5.1 Hypersensitivity Reactions 5.2 Injection Site Bruising 5.3 Substitution of Collagenase Products 6 Adverse Reactions 6.1 Clinical Trial Experience 6.2 Immunogenicity 6.3 Post - marketing Experience 8 Use in Specific Populations 8.1 Pregnancy 8.2 Lactation 8.4 Pediatric Use 8.5 Geriatric Use 11 Description 12 Clinical Pharmacology 12.1 Mechanism of Action 12.2 Pharmacodynamics 12.3 Pharmacokinetics 13 Non - clinical Toxicology 13.1 Carcinogenesis, Mutagenesis, Impairment of Fertility 14 Clinical Trials 16 How to Supply / Store and Handle 17 Patient Counseling Information * Sections or sub - sections omitted from sufficient prescribing information are not listed. Sufficient Prescribing Information 1 Indications and Usage QWO is indicated for the treatment of moderate to severe cellulite in the buttocks of adult women. 2 Dosage and Administration 2.1 Dosage QWO is administered by subcutaneous injection at a dose of 0.84 mg per treatment area. ● The treatment area is defined as a single buttock that receives up to 12 injections (up to a total of 3.6 mL) of 0.3 mL each of QWO. ● The treatment visit can consist of up to 2 treatment areas. Treatment should be repeated every 21 days during 3 treatment visits. 2.2 Reconstitution of Lyophilized Powder Before reconstitution, remove the vial from the freezer and allow it to stand at room temperature for at least 15 minutes. Inspect the vial containing QWO. The cake of lyophilized powder should be white and intact and show no signs of corrosion. The diluent should be a colorless solution free of particulate matter. After removing the flip-off cap(s) from the vial(s), using aseptic technique, wipe the rubber stopper and the surrounding surface of the vial(s) containing QWO and the diluent with sterile alcohol (no other bactericidal agent should be used). Use only the supplied diluent for reconstitution of QWO. Using a syringe and needle of appropriate size (not supplied), withdraw the amount of diluent supplied based on the number of injection sites (see Table 1). [Table 7] Slowly inject the diluent onto the surface of the vial containing the lyophilized powder of QWO. Do not invert the vial or shake the solution. Slowly rotate the solution to ensure that all of the lyophilized powder is dissolved. The reconstituted QWO solution in the vial can be maintained at room temperature (20°C to 25°C / 68°F to 77°F) for up to 8 hours or cooled at 2°C to 8°C (36°F to 46°F) for up to 72 hours before administration. If the reconstituted QWO solution in the vial is cooled, allow this solution to return to room temperature for approximately 15 minutes before use. The reconstituted QWO solution should be colorless, clear, and free of particulate matter. Visually inspect the solution for particulate matter or discoloration before administration. If the reconstituted QWO is not a colorless, clear solution essentially free of particulate matter, do not inject it. Discard the syringe(s) and needle(s) used for reconstitution and the diluent vial(s). After reconstitution, the QWO solution in the vial should be used for only one injection session and for only one patient. 2.3 Administration Preparation of Syringe for Injection Using a 1 mL syringe with a removable needle (not supplied), withdraw 0.9 mL of the reconstituted solution into each syringe. Refer to Table 2 for the appropriate number of syringes required based on the number of injection sites. After preparing the syringe, withdraw the solution remaining in the needle into the barrel of the syringe, then replace the needle with a 30-gauge 1 / 2-inch needle. The reconstituted solution prepared in the 1 mL syringe should be administered immediately. The reconstituted solution should not be stored in the 1 mL syringe. [Table 8] Injection Technique While the patient is standing, mark the injection sites. Inject QWO subcutaneously while the patient is in a bent-over position. Each injection of QWO should be administered as three 0.1 mL aliquots (total injection volume of 0.3 mL) at locations A, B, and C as shown in the figure below. Without downward pressure, the depth of the injection should be 0.5 inches (corresponding to the length of the needle). [Table 9] Needle Tip Position A: Position the needle at a 90° angle perpendicular to the skin surface at the injection site and slowly push the syringe plunger to inject one 0.1 mL aliquot. Needle Tip Position B: Withdraw the needle slightly (not so far as to come out of the injection site), reposition it at approximately 45° (not greater than 45°), and inject one 0.1 mL aliquot (towards the head). Needle Tip Position C: Withdraw the needle slightly (not so far as to come out of the injection site), reposition it at approximately 45° (not greater than 45°), and inject one 0.1 mL aliquot (towards the foot). Completely withdraw the needle from the skin and move to the next identified injection site. Each treatment area can receive up to 12 injections. After treatment, the patient should remain bent over for at least 5 minutes. After administration, do not store, pool, or use any vials or syringes containing the reconstituted solution that was not used. Discard any unused portions. 3 Dosage Forms and Strengths For injection: 0.92 mg or 1.84 mg of collagenase Clostridium histolyticum - aaes (seen as a white mass) as a lyophilized powder in a single - dose vial. 4 Contraindications QWO is contraindicated in: ● Patients with a history of hypersensitivity to either collagenase or an excipient [see Warnings and Precautions (5.1)] ● The presence of an infection at the injection site as indicated below. 5 Warnings and Precautions 5.1 Hypersensitivity Reactions Serious hypersensitivity reactions, such as anaphylaxis, have been reported with the use of collagenase Clostridium histolyticum. If such a reaction occurs, further injections of QWO should be discontinued and appropriate medical treatment should be initiated immediately. 5.2 Injection Site Bruising In clinical trials, 84% of subjects treated with QWO experienced injection site bruising [see Adverse Reactions (6.1)]. Subjects with coagulation disorders or those using anticoagulants or anti - platelet medications (except those taking ≤150 mg of aspirin daily) were excluded from participants in Trials 1 and 2. QWO should be used with caution in patients with bleeding disorders or those currently being treated with anti - platelet (except those taking ≤150 mg of aspirin daily) or anticoagulant therapy. 5.3 Substitution of Collagenase Products QWO should not be substituted with other injectable collagenase products. QWO is not intended for the treatment of Peyronie's disease or Dupuytren's contracture. 6 Adverse Reactions The following adverse reactions to QWO for injection: ● Hypersensitivity [see Contraindications (4) and Warnings and Precautions (5.1)] ● Bruising at the injection site (see Warnings and Precautions (5.2)) is discussed in more detail in other sections of the label. 6.1 Clinical Trial Experience Clinical trials are conducted under widely varying conditions, and the rates of adverse reactions 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 observed in clinical practice. In two double-blind, placebo-controlled clinical trials of the same design (Trials 1 and 2), 424 female subjects with cellulite on the buttocks received QWO and 419 female subjects with cellulite received placebo. Enrolled subjects were adults 18 to 78 years of age with moderate to severe cellulite (graded 3 or 4 on a scale of 0 to 4) and no excessive skin laxity. Most were white (78%) or African American (18%). Subjects were separated 21 days and completed up to 3 treatment visits and were followed for up to 6 months after the last treatment visit in another open-label extension trial (Trial 3). Table 3 shows the incidence of adverse reactions reported in ≥1% of subjects who received QWO at a higher frequency than subjects who received placebo in Trials 1 and 2 through Day 71. Generally, adverse reactions had a duration of less than 21 days.

Table 10

Table 11

Table 12

[0126] One of ordinary skill in the art will understand that many changes and modifications can be made to the preferred embodiments disclosed herein and that such changes and modifications can be made without departing from the spirit of the invention. Accordingly, the appended claims are intended to cover all such equivalent variations as fall within the true spirit and scope of the invention.

[0127] The disclosure of each patent, patent application, and publication cited or described in this document is hereby incorporated by reference in its entirety.

[0128] Aspects The following list of aspects is intended to complement, not replace or supersede, the previous description. Aspect 1. A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising: subcutaneously administering to a treatment area of the subject a total dose of collagenase of about 0.42 mg per treatment session, thereby reducing collagenase-mediated bruising in the subject. Aspect 2. The method of Aspect 1, wherein the collagenase is administered using a three-injection technique. Aspect 3. The method of Aspect 1 or 2, wherein the collagenase is administered at a depth of about 0.5 inches. Aspect 4. The method according to any one of Aspects 1 to 3, wherein the bruising is reduced as compared to the level of bruising associated with subcutaneous administration of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session for a total dose of about 0.84 mg to the treatment area of the subject. Aspect 5. The method according to any one of Aspects 1 to 4, wherein the collagenase has a concentration of about 0.23 mg / ml. Aspect 6. A method for reducing collagenase-mediated bruising in a subject having cellulite, 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. Aspect 7. The method according to Aspect 6, wherein the collagenase is administered using a three-injection technique. Aspect 8. The method according to Aspect 6 or 7, wherein the collagenase is administered at a depth of about 0.5 inches. Aspect 9. The method according to any one of Aspects 6 to 8, wherein the bruising is reduced as compared to the level of bruising associated with subcutaneous administration of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session for a total dose of about 0.84 mg to the treatment area of the subject, and the collagenase has a concentration of about 0.23 mg / ml. Aspect 10. A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising: subcutaneously administering to the treatment area of the subject a total dose of about 0.84 mg of collagenase at a depth of about 0.25 inches per treatment session, thereby reducing collagenase-mediated bruising in the subject. Aspect 11. The method according to Aspect 10, wherein the collagenase is administered using a three-injection technique. Aspect 12. The method according to Aspect 10 or 11, wherein the bruising is reduced as compared to the level of bruising associated with subcutaneous administration of collagenase at a depth of about 0.5 inches using a three-injection technique per treatment session for a total dose of about 0.84 mg to the treatment area of the subject. Aspect 13. The method according to any one of Aspects 10 to 12, wherein the collagenase has a concentration of about 0.23 mg / ml. Aspect 14. A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising: subcutaneously administering to a treatment area of the subject a total dose of about 0.84 mg of collagenase per treatment session, the collagenase having a concentration of about 0.09 mg / ml, thereby reducing collagenase-mediated bruising in the subject. Aspect 15. The method according to Aspect 14, wherein the collagenase is administered using a single aliquot injection. Aspect 16. The method according to Aspect 14 or 15, wherein the collagenase is administered at a depth of about 0.25 inches. Aspect 17. The method according to any one of Aspects 14 to 16, wherein the administering step comprises injecting a single aliquot of collagenase up to 30 per treatment session. Aspect 18. 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 at a depth of about 0.25 inches using a single aliquot injection per treatment session to a treatment area of the subject, and the collagenase has a concentration of about 0.23 mg / ml. The method according to any one of Aspects 14 to 17. Aspect 19. The method according to any one of Aspects 14 to 18, wherein the administering step comprises injecting a single aliquot of collagenase up to 12 per treatment session. Aspect 20. The method according to any one of Aspects 14 to 19, wherein the injections are spaced about 2 cm to about 3 cm apart. Aspect 21. The method according to Aspect 20, wherein the injections are spaced in a grid pattern. Aspect 22. A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine and epinephrine, wherein a total dose of about 0.42 mg of collagenase is administered per treatment session, thereby reducing collagenase-mediated bruising in the subject. Aspect 23. The method according to aspect 22, wherein the collagenase is administered using a three - injection technique. Aspect 24. The method according to aspect 22 or 23, wherein the collagenase is administered at a depth of about 0.5 inches. Aspect 25. The bruising is reduced compared to the level of bruising associated with subcutaneous administration of a composition containing collagenase to the treatment area of the subject, and a total dose of about 0.84 mg of collagenase per treatment session is administered at a depth of about 0.5 inches using a three - injection technique. The method according to any one of aspects 22 - 24. Aspect 26. The method according to any one of aspects 22 - 25, wherein the collagenase has a concentration of about 0.23 mg / ml. Aspect 27. The method according to any of the preceding aspects, wherein the subject undergoes a plurality of treatment sessions. Aspect 28. The method according to aspect 27, wherein the treatment sessions are separated by three weeks. Aspect 29. A method for reducing collagenase - mediated bruising in a subject having cellulite, comprising: subcutaneously administering to the treatment area of the subject a total dose of about 0.21 mg of collagenase per treatment session, wherein the collagenase has a concentration of about 0.12 mg / ml, thereby reducing collagenase - mediated bruising in the subject. Aspect 30. The method according to aspect 29, wherein the collagenase is administered using a three - injection technique. Aspect 31. The method according to aspect 29 or 30, wherein the collagenase is administered at a depth of about 0.5 inches. Aspect 32. The bruising is 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 using a three - injection technique per treatment session to the treatment area of the subject, and the collagenase has a concentration of about 0.23 mg / ml. The method according to any one of aspects 29 - 31. Aspect 33. The method according to any one of aspects 29 - 32, wherein the subject undergoes more than one treatment session separated by about 42 days. Aspect 34. A method for reducing collagenase - mediated bruising in a subject having cellulite, comprising: A method comprising, per treatment session, orally administering tranexamic acid to a subject and subcutaneously administering a total dose of about 0.84 mg of collagenase to a treatment area of the subject, thereby reducing collagenase-mediated bruising in the subject. Aspect 35. The method according to aspect 34, wherein the collagenase is administered using a three-injection technique. Aspect 36. The method according to aspect 34 or 35, wherein the collagenase is administered at a depth of about 0.5 inches. Aspect 37. The method according to any one of aspects 34 to 36, comprising administering about 500 mg to about 1500 mg of tranexamic acid to the subject. Aspect 38. The method according to aspect 37, comprising administering about 1300 mg of tranexamic acid to the subject. Aspect 39. The method according to any of the preceding aspects, wherein the collagenase is administered in a composition comprising collagenase, mannitol, sucrose and tromethamine. Aspect 40. The method according to any of the preceding aspects, wherein the level of bruising is evaluated using the Investigator Assessment of Bruising Severity Scale (IABSS). Aspect 41. a) Subcutaneously administering a total dose of about 0.42 mg of collagenase per treatment session to a treatment area of the subject, thereby treating cellulite in the subject; b) Subcutaneously administering a total dose of about 0.84 mg of collagenase per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.05 mg / ml, thereby treating cellulite in the subject; c) Subcutaneously administering a total dose of about 0.84 mg of collagenase per treatment session to a treatment area of the subject at a depth of about 0.25 inches, the collagenase having a concentration of about 0.23 mg / ml, thereby treating cellulite in the subject; d) Subcutaneously administering a total dose of about 0.84 mg of collagenase per treatment session to a treatment area of the subject, the collagenase having a concentration of about 0.09 mg / ml, thereby treating cellulite in the subject; e) administering subcutaneously to the treatment area of the subject a composition comprising collagenase, lidocaine and epinephrine, wherein a total dose of collagenase of about 0.42 mg per treatment session is administered, whereby cellulite is treated in the subject; f) administering subcutaneously to the treatment area of the subject a total dose of collagenase of about 0.21 mg per treatment session, the collagenase having a concentration of about 0.12 mg / ml, whereby cellulite is treated in the subject; or g) orally administering tranexamic acid to the subject per treatment session and subcutaneously administering to the treatment area of the subject a total dose of about 0.84 mg of collagenase, whereby cellulite is treated in the subject A method for treating cellulite in a subject, comprising: Aspect 42. The method according to aspect 41, wherein the collagenase is administered using a three - injection technique. Aspect 43. The method according to aspect 41, wherein the collagenase is administered using a single - aliquot injection. Aspect 44. The method according to aspect 42 or 43, wherein the collagenase is administered at a depth of about 0.25 inches or about 0.5 inches. Aspect 45. The method according to any one of aspects 41 - 44, wherein in step g), the tranexamic acid has a concentration of about 500 mg to about 1500 mg. Aspect 46. The method according to aspect 45, wherein in step g), the tranexamic acid has a concentration of about 1300 mg. Aspect 47. The method according to any one of aspects 41 - 46, wherein the collagenase is administered in a composition comprising collagenase, mannitol, sucrose and tromethamine. Aspect 48. The method according to aspect 41, wherein the collagenase in part a) has a concentration of about 0.23 mg / ml. Aspect 49. The method according to aspect 41, wherein the injections in part d) are spaced about 2 cm to about 3 cm apart. Aspect 50. The method according to aspect 49, wherein the injections are spaced in a grid pattern. Aspect 51. The method according to any one of aspects 41 - 50, wherein the subject undergoes a plurality of treatment sessions. Aspect 52. The method according to aspect 51, wherein the treatment sessions are separated by 3 weeks. Aspect 53. The method according to aspect 51, wherein the treatment sessions are separated by about 42 days. Aspect 54. The treatment is established by a scale or measurement tool selected from the Hexsel cellulite severity scale (Hexsel CSS), Hexsel depression depth score, recurrence scale, dimple analysis, clinician-reported photographic numerical cellulite severity scale (CR-PCSS), patient-reported photographic numerical 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), abbreviated PR-CIS, subject self-assessment scale (SSRS), subject satisfaction with cellulite treatment (SSCT), clinician assessment of cellulite severity (photograph or other image), Body-Q, and validated photographic numerical or other scales used by clinicians and / or patients to evaluate cellulite severity, improvement, and / or patient satisfaction. The method according to any one of aspects 41 to 53. Aspect 55. The method according to any of the preceding aspects, wherein the collagenase comprises collagenase I, collagenase II, or a mixture of collagenase I and collagenase II. Aspect 56. The method according to aspect 55, wherein the collagenase comprises a mixture of collagenase I and collagenase II. Aspect 57. The method according to aspect 56, 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. Aspect 58. The method according to any of the preceding aspects, wherein the collagenase is collagenase Clostridium histolyticum (CCH). Aspect 59. The method according to any of the preceding aspects, wherein the treatment area is the left buttock, the right buttock, or both the left and right buttocks. Aspect 60. Collagenase; Lidocaine; and Epinephrine A pharmaceutical composition comprising. Aspect 61. About 0.23 mg / ml of collagenase; About 2% lidocaine; and Epinephrine at about 1:200,000 The formulation according to embodiment 60, comprising Embodiment 62. The formulation according to embodiment 60 or 61, wherein the collagenase comprises collagenase I, collagenase II, or a mixture of collagenase I and collagenase II. Embodiment 63. The formulation according to embodiment 62, wherein the collagenase comprises a mixture of collagenase I and collagenase II. Embodiment 64. The formulation according to embodiment 63, 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. Embodiment 65. The formulation according to any one of embodiments 60 to 64, wherein the collagenase is collagenase Clostridium histolyticum (CCH).

Claims

**Claim 1** A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising subcutaneously administering a total dose of collagenase of about 0.42 mg per treatment session to a treatment area of the subject, thereby reducing collagenase-mediated bruising in the subject. **Claim 2** The method according to claim 1, wherein the collagenase is administered using a three-injection technique. **Claim 3** The method according to claim 1 or 2, wherein the collagenase is administered at a depth of about 0.5 inches. **Claim 4** The method according to any one of claims 1 to 3, wherein the collagenase has a concentration of about 0.23 mg / ml. **Claim 5** A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising subcutaneously administering a total dose of collagenase of about 0.84 mg per treatment session to a treatment area of the subject, wherein the collagenase has a concentration of about 0.05 mg / ml, thereby reducing collagenase-mediated bruising in the subject. **Claim 6** The method according to claim 5, wherein the collagenase is administered using a three-injection technique. **Claim 7** The method according to claim 5 or 6, wherein the collagenase is administered at a depth of about 0.5 inches. **Claim 8** A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising subcutaneously administering a total dose of collagenase of about 0.84 mg at 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. **Claim 9** The method according to claim 8, wherein the collagenase is administered using a three-injection technique. **Claim 10** The method according to claim 8 or 9, wherein the collagenase has a concentration of about 0.23 mg / ml. **Claim 11** A method for reducing collagenase-mediated bruising in a subject having cellulite, comprising subcutaneously administering a total dose of collagenase of about 0.84 mg per treatment session to a treatment area of the subject, wherein the collagenase has a concentration of about 0.09 mg / ml, thereby reducing collagenase-mediated bruising in the subject. **Claim 12** The method according to claim 11, wherein the collagenase is administered using a single-aliquot injection. **Claim 13** The method according to claim 11 or 12, wherein the collagenase is administered at a depth of about 0.25 inches. **Claim 14** The method according to any one of claims 11 to 13, wherein the administration step comprises injection of a single aliquot of collagenase up to 30 per treatment session.

15. The method according to any one of claims 11 to 13, wherein the administration step comprises injection of a single aliquot of collagenase up to 12 per treatment session.

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

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

18. A method for reducing collagenase-mediated bruising in a subject having cellulite, the method comprising the step of subcutaneously administering to a treatment area of the subject a composition comprising collagenase, lidocaine, and epinephrine, wherein: A method in which a total dose of about 0.42 mg of collagenase is administered per treatment session, thereby reducing collagenase-mediated bruising in the subject.

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

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

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

22. The method according to any of the preceding claims, wherein the subject undergoes a plurality of treatment sessions.

23. The method according to claim 22, wherein the treatment sessions are spaced 3 weeks apart.

24. A method for 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 about 0.21 mg of collagenase per treatment session, wherein the collagenase has a concentration of about 0.12 mg / ml, thereby reducing collagenase-mediated bruising in the subject.

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

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

27. The method according to any one of claims 24 to 26, wherein the subject undergoes more than one treatment session spaced about 42 days apart.

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

29. The method according to any of the preceding claims, wherein the level of bruising is evaluated using the Investigator's Assessment of Bruising Severity Scale (IABSS).

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

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

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

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

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

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

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

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

38. The method according to any one of claims 30 to 37, wherein the subject undergoes a plurality of treatment sessions.

39. The method according to claim 38, wherein the treatment sessions are separated by 3 weeks.

40. The method according to claim 38, wherein the treatment sessions are separated by about 42 days.

41. The treatment is established by a scale or measurement tool selected from the Hexsel cellulite severity scale (Hexsel CSS), Hexsel depression depth score, Ricart scale, pit analysis, clinician-reported photographic numerical cellulite severity scale (CR-PCSS), patient-reported photographic numerical 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), abbreviated PR-CIS, subject self-assessment scale (SSRS), subject satisfaction with cellulite treatment (SSCT), clinician assessment of cellulite severity (photograph or other image), Body-Q, and validated photographic numerical or other scales used by clinicians and / or patients to evaluate cellulite severity, improvement, and / or patient satisfaction, the method according to any one of claims 30 to 40.

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

44. The method according to claim 43, 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.

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

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

47. Collagenase; Lidocaine; and Epinephrine A pharmaceutical formulation comprising.

48. About 0.23 mg / ml of collagenase; About 2% lidocaine; and About 1:200,000 epinephrine The pharmaceutical formulation according to claim 47, comprising.

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

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

51. The preparation according to claim 50, 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.

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

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