Injectable compositions for reducing fat, comprising a cell-lysating compound in a gel, gel-forming solution, or gel-forming suspension.

Abstract

The present invention provides an injectable composition, comprising a cytolytic compound, preferably deoxycholic acid or its salt, more preferably DCA-Na, as a first component, and a pharma-ceutically acceptable excipient.The present invention also provides the use of the injectable composition for reducing or removing local fat in a subject in need of reducing or removing local fat, the injectable composition being subcutaneously injected into a subcutaneous injection site of the subject.The present invention also provides a method for reducing or removing local fat in a subject in need of reducing or removing local fat, comprising administering an effective amount of the injectable composition to the subject.Specifically, the injectable composition of the present invention can be in the form of a gel.

Description

[Technical Field]

[0001] The present invention relates to the preparation of an injectable composition. More specifically, the present invention relates to an injectable composition for reducing fat, comprising a cell-lytic compound in a gel, gel-forming solution, or gel-forming suspension, and to the use or method for reducing or removing local fat by administering the injectable composition of the present invention. Specifically, the injectable composition of the present invention may be in the form of a gel during or after injection. [Background technology]

[0002] Because submandibular fat and double chins typically resist diet or exercise, non-surgical fat removal injections using the active ingredient deoxycholic acid are becoming a new treatment option for reducing submandibular fat.

[0003] Deoxycholic acid (DCA) is a secondary bile acid that can emulsify and solubilize fats for digestion and absorption in the intestines. Its salt, sodium deoxycholate (DCA-Na), is an anionic surfactant commonly used to lyse cells. DCA is a TGR5 agonist (Takeda G protein-coupled receptor 5, GPBAR1), and activation of TGR5 has been shown to reduce obesity in animals fed a high-fat diet. DCA is predicted to lyse adipocytes, leading to fat reduction. However, cell lysis attracts inflammatory cells such as macrophages and monocytes to remove the destroyed adipocytes. Patients treated with deoxycholic acid often experience swelling, pain, numbness, redness, and induration at the time of treatment due to inflammation, and the interval between treatments is long (about one month), as histological evidence has shown that most post-treatment inflammation has resolved by this time. DCA-Na can form hydrogels when mixed with tris(hydroxymethyl)aminomethane (TRIS) buffer at low pH, or when mixed with the polymer and amino acid L-aspartic acid. Since the release of additional solutes on DCA-Na / TRIS hydrogels has been found to be sustained, it should be a suitable drug delivery and release platform. While some studies have shown that adding the amino acids L-lysine and L-arginine, rather than glycine and L-α-alanine, weakens their hydrogel formation, we have successfully constructed DCA-Na gel systems by mixing with basic amino acids such as L-lysine, L-arginine, and L-histidine, as well as / or organic acids such as acetic acid.

[0004] Studies have shown that after injection of deoxycholic acid solution, deoxycholic acid penetrates into adipose tissue exceeding 1 centimeter. Adipose tissue cells larger than 2 centimeters in diameter trigger an inflammatory response. When deoxycholic acid gel solution is injected into adipose tissue, only the adipocytes surrounding the deoxycholic acid gel are gradually destroyed during the 7-day sustained release of deoxycholic acid. The inflammatory response is limited to a thin layer of adipocytes less than 2 millimeters thick surrounding the deoxycholic acid gel. The total volume of inflammatory adipose tissue is less than 10% of that of conventional cell-lysing injections. Finally, cavities of a volume proportional to the amount of deoxycholic acid injected appear in the adipose tissue and disappear within 2-3 weeks.

[0005] Therefore, a sustained-release deoxycholic acid, or sodium deoxycholate (DCA-Na) gel, is desired at the injection site, constructed by mixing it with basic amino acids such as L-lysine, L-arginine, and L-histidine, and / or organic acids such as acetic acid, so that the cell lysis reaction can be limited to adipocytes infiltrated by deoxycholic acid around the gel surface. Furthermore, anti-inflammatory drugs or local anesthetics may be added to the injection during treatment to reduce inflammation and pain. The inventors also aimed to increase the concentration of DCA-Na so that cell lysis is more effective, allowing patients to complete treatment in fewer sessions. In summary, a mixture of DCA-Na, basic amino acids, and / or organic acids with anti-inflammatory drugs and / or local anesthetics should reduce or remove fat, effectively mitigate adverse effects, shorten the interval between treatments, and shorten the overall treatment process. The DCA-Na injection composition preferably forms a gel-like appearance between 5 and 120 minutes after mixing. [Overview of the project]

[0006] The present invention provides an injectable composition of a cell-lysating compound, preferably deoxycholic acid or a salt thereof, more preferably DCA-Na, in the form of a gel, gel-forming solution, or gel-forming suspension. This injectable composition can be used for local fat reduction or removal, has few side effects, and the treatment process is relatively short.

[0007] In one embodiment, the present invention provides an injectable composition for reducing fat, comprising a cell-lysing compound in a gel, gel-forming solution, or gel-forming suspension, the injectable composition, The first component is a cell-lytic compound, and Contains pharmaceutically acceptable excipients.

[0008] Preferably, the cell-lysating compound is deoxycholic acid or a salt thereof.

[0009] More preferably, the cell-lysating compound is DCA-Na, and the injectable composition further comprises a second component selected from one or more basic amino acids or organic acids.

[0010] In some embodiments, the concentration of DCA-Na is 7 to 51 mg / mL.

[0011] In some embodiments, the basic amino acid is L-lysine.

[0012] In one embodiment, the concentration of L-lysine is 11 to 145 mg / mL.

[0013] In another embodiment, the pH of the L-lysine before mixing is less than 8.0, and the pH of the injectable composition is 6.45 to 7.75.

[0014] In another embodiment, the injectable composition further comprises an anti-inflammatory agent as a third component.

[0015] Preferably, the anti-inflammatory drug is aspirin.

[0016] More preferably, the aspirin concentration is 14-100 mg / mL.

[0017] Preferably, the injectable composition further comprises a local anesthetic as a fourth component.

[0018] More preferably, the local anesthetic is lidocaine.

[0019] More preferably, the concentration of lidocaine is 2.5 - 6.5 mg / mL.

[0020] Preferably, the anti - inflammatory drug is dexamethasone sodium phosphate (DSP).

[0021] More preferably, the pH of the injectable composition is 6.45 - 7.40.

[0022] More preferably, the concentration of DSP is 1 mg / mL or less.

[0023] In some embodiments, the basic amino acid is L - histidine.

[0024] Preferably, the concentration of L - histidine is 1.4 - 11.5 mg / mL.

[0025] In some embodiments, the basic amino acid is L - arginine.

[0026] Preferably, the concentration of L - arginine is 115 - 143 mg / mL.

[0027] In some embodiments, the organic acid is acetic acid.

[0028] Preferably, the concentration of acetic acid is 46 - 143×10 -3 %.

[0029] In other embodiments, the injectable composition further contains physiological saline.

[0030] In other embodiments, the injectable composition is preferably in the form of a gel during and after injection.

[0031] In another embodiment, the present invention provides the use of the above-described injectable composition for reducing or removing local fat in subjects requiring reduction or removal of local fat, wherein the injectable composition is subcutaneously injected into the subcutaneous injection site of the subject.

[0032] In another embodiment, the subcutaneous injection site is local fat within the face, chin, arm, waist, abdomen, or thigh of the subject.

[0033] In another embodiment, the present invention provides the use of the above-mentioned injectable composition for the manufacture of a pharmaceutical for the reduction or removal of local fat.

[0034] In another embodiment, the present invention provides a method for reducing or removing local fat in a subject requiring reduction or removal of local fat, comprising administering an effective amount of the injectable composition to the subject, preferably by subcutaneous injection.

[0035] In another embodiment, the subject is a human.

[0036] In another embodiment, the injectable composition is administered, preferably by subcutaneous injection, to localized fat tissue in the face, chin, arms, waist, abdomen, or thighs of the target area.

[0037] The injectable composition of the present invention may also contain physiological saline and may be in the form of a gel during or after injection. [Brief explanation of the drawing]

[0038] [Figure 1-1] The images show the appearance of a mixture of DCA-Na solution and L-lysine solution at (a) 100 mg / mL, (b) 200 mg / mL, (c) 300 mg / mL, (d) 400 mg / mL, or (e) 500 mg / mL concentrations. [Figure 1-2] The images show the appearance of a mixture of DCA-Na solution and L-lysine solution at (a) 100 mg / mL, (b) 200 mg / mL, (c) 300 mg / mL, (d) 400 mg / mL, or (e) 500 mg / mL concentrations. [Figure 2]The images show the appearance of DCA-Na solutions at various pH levels and mixtures of (a) 200 mg / mL or (b) 400 mg / mL L-lysine solutions. [Figure 3-1] The images show the appearance of a mixture of DCA-Na solution and LA at (a) 90 mg / mL, (b) 180 mg / mL, (c) 300 mg / mL, (d) 450 mg / mL, or (e) 600 mg / mL concentrations. [Figure 3-2] The images show the appearance of a mixture of DCA-Na solution and LA at (a) 90 mg / mL, (b) 180 mg / mL, (c) 300 mg / mL, (d) 450 mg / mL, or (e) 600 mg / mL concentrations. [Figure 4-1] This shows the appearance of a mixture of CA-Na solution and LA in lidocaine HCl solution at concentrations of (a) 90 mg / mL, (b) 180 mg / mL, (c) 300 mg / mL, (d) 450 mg / mL, or (e) 600 mg / mL. [Figure 4-2] This shows the appearance of a mixture of CA-Na solution and LA in lidocaine HCl solution at concentrations of (a) 90 mg / mL, (b) 180 mg / mL, (c) 300 mg / mL, (d) 450 mg / mL, or (e) 600 mg / mL. [Figure 5] These are photographs of adipose tissue collected from both sides (L: left side, R: right side) of two pigs, with (a) and (b) from the first pig and (c) and (d) from the second pig. [Figure 6] The images show the appearance of DCA-Na solutions at various pH levels and mixtures of (a) 200 mg / mL or (b) 400 mg / mL L-lysine / DSP solutions. [Figure 7-1] These are photographs of adipose tissue collected from both sides (L: left side, R: right side) of three pigs, with (a) and (b) from the first pig, (c) and (d) from the second pig, and (e) and (f) from the third pig. [Figure 7-2] These are photographs of adipose tissue collected from both sides (L: left side, R: right side) of three pigs, with (a) and (b) from the first pig, (c) and (d) from the second pig, and (e) and (f) from the third pig. [Figure 8-1] This shows the appearance of a mixture of DCA-Na solution and L-histidine solutions at concentrations of (a) 2.5 mg / mL, (b) 5 mg / mL, (c) 10 mg / mL, (d) 20 mg / mL, (e) 40 mg / mL, or (f) 50 mg / mL. [Figure 8-2] This shows the appearance of a mixture of DCA-Na solution and L-histidine solutions at concentrations of (a) 2.5 mg / mL, (b) 5 mg / mL, (c) 10 mg / mL, (d) 20 mg / mL, (e) 40 mg / mL, or (f) 50 mg / mL. [Figure 9] This is the appearance of a mixture of DCA-Na solution and 500 mg / mL L-arginine solution. [Figure 10-1] The images show the appearance of a mixture of DCA-Na solution and (a) 0.1%, (b) 0.2%, (c) 0.3%, (d) 0.4%, (e) 0.5%, or (f) 0.6% L-histidine solution. [Figure 10-2] The images show the appearance of a mixture of DCA-Na solution and (a) 0.1%, (b) 0.2%, (c) 0.3%, (d) 0.4%, (e) 0.5%, or (f) 0.6% L-histidine solution. [Figure 10-3] The images show the appearance of a mixture of DCA-Na solution and (a) 0.1%, (b) 0.2%, (c) 0.3%, (d) 0.4%, (e) 0.5%, or (f) 0.6% L-histidine solution. [Modes for carrying out the invention]

[0039] definition In the present invention, the following definitions are applicable.

[0040] In this invention, the articles "a" and "an" are used to refer to one or more (i.e., at least one) grammatical objects of the articles. For example, "an element" means one or more elements.

[0041] In this invention, the term "and / or" is used to mean either "and" or "or" unless otherwise indicated.

[0042] The term "effective amount" means an amount of the composition according to the present invention that is sufficient to achieve the desired effect or result in the given administration or use of the composition. The effective amount can be determined by methods known to those skilled in the art.

[0043] The "subject" is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cattle, pig, or a non-human primate, such as a monkey, chimpanzee, baboon, or rhesus macaque. The subject of the present invention is preferably a human.

[0044] The term "pharmaceutically acceptable excipients," as used herein, refers to compounds that are generally safe, non-toxic, and not biologically or otherwise undesirable, and are useful for preparing pharmaceutical compositions, and includes excipients acceptable for veterinary or human pharmaceutical use. The pharmaceutically acceptable excipients as used herein and in the claims include both one and more such excipients. Suitable excipients include solvents such as sterile water or water for injection; lubricants such as talc and magnesium stearate; wetting agents, emulsifiers and suspending agents; isotonic agents such as sodium chloride; acids such as hydrochloric acid; bases such as sodium hydroxide; buffers such as disodium hydrogen phosphate; and preservatives such as methyl benzoate, propyl hydroxybenzoate and benzyl alcohol.

[0045] The "cell-lysing compound" may also be a surfactant or a lipolytic compound. Suitable cell-lysing compounds include, but are not limited to, phosphatidylcholine, deoxycholic acid, or salts thereof. The cell-lysing compound of the present invention is preferably deoxycholic acid or a salt thereof, and more preferably DCA-Na.

[0046] Aspirin (acetylsalicylic acid) is a nonsteroidal anti-inflammatory drug (NSAID) used to relieve pain, fever, or inflammation, but it also suppresses normal platelet function. Its soluble salt, lysine aspirin (LA), can be administered intravenously or intramuscularly. After administration, lysine aspirin is converted to acetylsalicylic acid and metabolized to salicylic acid.

[0047] Dexamethasone is a glucocorticosteroid similar to a natural hormone produced by the adrenal glands. Dexamethasone relieves inflammation (swelling, fever, redness, and pain) and is used to treat certain types of arthritis, severe allergies, asthma, and certain types of cancer. Dexamethasone sodium phosphate (DSP) is a form of its sodium phosphate salt.

[0048] Lidocaine (or lignocaine) is an aminoamide-type local anesthetic that can temporarily block the transmission of nerve impulses. Lidocaine typically begins to take effect within minutes of administration and lasts for 30 minutes to 3 hours. Lidocaine mixtures can also be applied directly to the skin or mucous membranes to numb the area. [Examples]

[0049] The present invention can be better understood by following the following examples. However, it will be readily apparent to those skilled in the art that the examples are not intended to limit the present invention as described in detail in the claims, but merely to illustrate the present invention. Unless otherwise indicated, the compositions of the present invention can be prepared using commercially available materials and by utilizing general techniques and procedures known to those skilled in the art.

[0050] DCA-Na solution DCA-Na (99%, Acros Organics, Geel, Belgium), NaOH, Na2HPO4 (Sigma-Aldrich, St. Louis, MO, USA), and NaCl (Honeywell, Charlotte, NC, USA) were added to 80 mL of sterile water for injection, and then the solution was made to 100 mL. Benzyl alcohol (Alfa Aesar, Ward Hill, MA, USA) was then added to the solution, and the pH was adjusted by adding additional sodium hydroxide / hydrochloric acid. The amounts and concentrations of each component are shown in Tables 1 and 2, and 5% and 1% solutions were prepared, respectively. The solutions were sterilized by autoclaving for 30 minutes.

[0051] [Table 1]

[0052] [Table 2]

[0053] In the following examples, injectable compositions were prepared by mixing DCA-Na solution with other components. Unless otherwise specified, the final concentration of DCA-Na in the resulting composition was required to be 70% or more of the initial solution (36.96 mg / mL or more for a 5% solution, and 7.39 mg / mL or more for a 1% solution). The appearance of the mixtures after mixing DCA-Na with other components was observed after standing at 25°C, 37°C, and 42°C for 20 minutes, 30 minutes, 45 minutes, 60 minutes, and 120 minutes. In addition, 200 μL of the mixture was added to 200 μL of 0.9% physiological saline, and the appearance of the mixtures was also observed after standing at 37°C for 20 minutes, 30 minutes, 45 minutes, 60 minutes, and 120 minutes. Photographs were taken and shown in the figure.

[0054] Example 1. Composition of DCA-Na and L-lysine To investigate whether the DCA-Na and L-lysine compositions form a gel after mixing, DCA-Na solutions were mixed with acidic L-lysine solutions (pH 5.0-5.2, Acros Organics) according to Table 3.

[0055] [Table 3-1]

[0056] [Table 3-2]

[0057] [Table 3-3]

[0058] Figure 1 shows that when lysine solution was added to DCA-Na solution, all groups formed a clear solution. Mixtures of DCA-Na and lysine with higher concentrations of lysine (Figure 1, 1c-1e) began to form a gel after about 30 minutes when placed at 25°C (remaining at the bottom of the bottle after inversion), but the DCA-Na and lysine mixtures placed at 42°C did not form a gel at all lysine concentrations in the tested 5% DCA-Na and lysine concentrations below 140 mg / mL in 1% DCA-Na. Mixtures of 5% DCA-Na and lysine added to 0.9% physiological saline formed a gel after about 60 minutes when the lysine concentration was greater than 83 mg / mL and after about 30 minutes when the lysine concentration was greater than 85 mg / mL (Figure 1, 1c-1e). A mixture of 1% DCA-Na and lysine added to 0.9% physiological saline formed a gel in approximately 60 minutes when the lysine concentration was greater than 45 mg / mL, in approximately 30 minutes when the lysine concentration was greater than 69 mg / mL, and in approximately 20 minutes when the lysine concentration was greater than 85 mg / mL (Figure 1, 1c-1e). These results indicate that higher concentrations of LA formed a gel in a shorter time. Therefore, it is suggested that the mixture of DCA-Na and lysine should be used as soon as possible after mixing.

[0059] In Example 1, when the final concentration of DCA-Na was 7.54 to 44.00 mg / mL and the final concentration of L-lysine was 45.45 to 142.86 mg / mL, the composition to which 0.9% physiological saline was added was able to form a gel.

[0060] To determine the optimal pH values ​​for DCA-Na solution and L-lysine solution that form a gel after mixing, DCA-Na solution was mixed with L-lysine at various pH levels according to Table 4.

[0061] [Table 4-1]

[0062] [Table 4-2]

[0063] [Table 5]

[0064] Figure 2 shows that when lysine solution was added to DCA-Na solution, all groups formed a clear solution. In 5% and 1% DCA-Na solutions mixed with 200 mg / mL L-lysine solution with a pH in the range of 4.0 to 10.0, the pH values ​​of the mixed solutions were in the range of 7.21 to 9.97 and 6.71 to 9.92, respectively. In 5% and 1% DCA-Na solutions mixed with 400 mg / mL L-lysine solution with a pH in the range of 5.0 to 10.0, the pH values ​​were in the range of 7.45 to 9.92 and 6.96 to 9.89 (Table 5). In the 200 mg / mL L-lysine test, 5% DCA-Na and L-lysine added to 0.9% physiological saline formed a gel in approximately 60 minutes at pH 4.0, while a mixture of 1% DCA-Na and L-lysine added to 0.9% physiological saline formed a gel in approximately 30 minutes at pH 4.0 and approximately 45 minutes at pH 5.0 (Figure 2, 2a). In the 400 mg / mL L-lysine test, a mixture of 5% DCA-Na and L-lysine added to 0.9% physiological saline formed a gel in approximately 45 minutes at pH 5.0 and 6.0 and approximately 60 minutes at pH 7.0, while a mixture of 1% DCA-Na and L-lysine added to 0.9% physiological saline formed a gel in approximately 30 minutes at pH 5.0 and approximately 45 minutes at pH 6.0 (Figure 2, 2b). Therefore, the preferred pH of the L-lysine solution before mixing is less than 8.0, preferably 5.0 to 7.0, and more preferably around pH 5.0 to 6.0. It has been suggested that the pH decreases as the concentration of L-lysine decreases.

[0065] A composition to which 0.9% physiological saline has been added can form a gel when the final pH of the composition is between 7.02 and 7.70.

[0066] Example 2. Composition of DCA-Na and lysine aspirin To test whether a gel could be formed by mixing DCA-Na solution with a lysine-containing NSAID, DCA-Na solution was mixed with LA (Lyacety, 0.9g / bottle, equivalent to 0.5g aspirin, China Chemical & Pharmaceutical Co., Ltd., Taipei, China) according to Table 6.

[0067] [Table 6-1]

[0068] [Table 6-2]

[0069] [Table 6-3]

[0070] Figure 3 shows that when the LA solution was added to the DCA-Na solution, all groups formed a clear solution. Mixtures of DCA-Na and LA with higher concentrations began to form a gel in about 20 minutes when left at 25°C (Figure 3, 3d, 3e), while mixtures left at 37 or 42°C took longer to form a gel but formed a suspension (or precipitate) in a shorter time (Figure 3, 3b-3e). When added to 0.9% physiological saline, mixtures formed a gel in about 60 minutes when the LA concentration was 50 mg / mL or higher, and in about 30 minutes when the LA concentration exceeded 69 mg / mL (Figure 3, 3b-3e). Higher LA concentrations resulted in faster gel formation. Therefore, it is suggested that mixtures of DCA-Na and LA should be used as soon as possible after mixing.

[0071] In Example 2, when the final concentration of DCA-Na was 7.54 to 48.00 mg / mL, with a maximum of 50.29 mg / mL, and the final concentration of LA was 25.71 to 171.43 mg / mL, the composition with 0.9% physiological saline added was able to form a gel, where the final concentrations of lysine and aspirin were approximately 11.40 to 76.81 mg / mL and 14.31 to 94.62 mg / mL, respectively.

[0072] Example 3. Lysine aspirin composition with DCA-Na and lidocaine HCl added. To test whether the DCA-Na solution and the LA dissolved in the local anesthetic lidocaine HCl formed a gel after mixing, the DCA-Na solution was mixed with LA in lidocaine HCl (5 mL / bottle, Lita Pharmacy CO., Ltd., Taichung City, China) according to Table 7.

[0073] [Table 7-1]

[0074] [Table 7-2]

[0075] [Table 7-3]

[0076] Figure 4 shows that when LA in lidocaine HCl solution was added to DCA-Na solution, all groups formed a clear solution. Mixtures of DCA-Na and LA in lidocaine HCl with a high concentration of LA began to form a gel in about 30 minutes when left at 25°C (Figure 4, 4e), while mixtures left at 37 or 42°C took longer to form a gel but formed a suspension or precipitate in a shorter time (Figure 4, 4a-4e). In the case of 5% DCA-Na solution, mixtures of DCA-Na and LA in lidocaine HCl added to 0.9% physiological saline formed a gel in about 60 minutes when the LA concentration exceeded 70 mg / mL, about 45 minutes when the LA concentration exceeded 134 mg / mL, and about 30 minutes when the LA concentration exceeded 170 mg / mL (Figure 4, 4c-4e). The concentration of lidocaine HCl to be mixed with 5% DCA-Na solution was acceptable up to 6 mg / mL. In the case of a 1% DCA-Na solution, a mixture of DCA-Na and LA + lidocaine HCl added to 0.9% physiological saline formed a gel in approximately 120 minutes when the LA concentration exceeded 40 mg / mL, approximately 60 minutes when the LA concentration exceeded 67 mg / mL, and approximately 45 minutes when the LA concentration exceeded 85 mg / mL (Figure 4, 4b-4e). The optimal concentration of lidocaine HCl to be mixed with the 1% DCA-Na solution was approximately 3 mg / mL. These results indicate that precipitation is likely to occur when a high concentration of lidocaine is added to a low concentration of DCA-Na.

[0077] In Example 3, when the final concentration of DCA-Na was 8.12–44.90 mg / mL and the final concentration of LA was 41.54–179.83 mg / mL, the composition to which 0.9% physiological saline was added was able to form a gel. Here, the final concentrations of lysine and aspirin were approximately 18.61–80.61 mg / mL and 22.93–99.22 mg / mL, respectively, and the final concentration of lidocaine was 2.99–6.99 mg / mL.

[0078] Example 4. Effects of lysine aspirin with DCA-Na and lidocaine HCl in porcine tissue. Two SPF Landrace male pigs, approximately 5-6 months old, were anesthetized by intramuscular injection of 0.04 mg / kg atropine. 10-15 minutes later, 6 mg / kg Zoletil 50 and 2.2 mg / kg Rompun were administered intramuscularly. 1.5 mL of lidocaine HCl was added to the LA and mixed until dissolved. 0.35 mL of the lidocaine HCl / LA solution was added to 2 mL of 1% or 5% DCA-Na solution and mixed until dissolved. The pigs were injected with 0.9% saline, 1% or 5% DCA-Na solution, with or without the addition of lidocaine HCl / LA, at different time points according to Table 8. The area of ​​each injection site was 16 cm². 2 The composition was injected at a depth of 1.0 cm in the center of each site. 55 injection sites were administered to each side of the pig (110 sites in total per pig). After slaughter (day 0), adipose tissue samples were taken and cut in half from the center. Photographs of the sections were recorded and shown in Figure 5.

[0079] [Table 8]

[0080] As shown in Figure 5, seven days after injection, the injection sites for DCA-Na solution alone were slightly harder and considerably more swollen than the injection sites for DCA-Na solution containing lidocaine HCl / LA. Figure 5 shows that cell lysis occurred at the injection site when DCA-Na solution was injected alone (groups 4-7). On the other hand, when DCA-Na solution was injected together with lidocaine HCl / LA, cell lysis occurred at the bottom of the adipose tissue (groups 8-15). This may suggest that only DCA-Na solution tends to diffuse into the adipose tissue; however, when DCA-Na solution is mixed with lidocaine HCl / LA, a gel is formed that can be deposited and diffused at the bottom of the adipose tissue, which is consistent with a decrease in hardness on palpation. Cell lysis and / or inflammation were observable at the injection sites for DCA-Na solution alone for at least 21-28 days, but were difficult to observe at the injection sites for DCA-Na solution containing lidocaine HCl / LA after 21 days.

[0081] A composition containing lidocaine HCl, DCA-Na, and lysine aspirin can effectively reduce fat with fewer side effects such as inflammation.

[0082] Example 5. Composition of L-lysine containing DCA-Na and DSP To test whether a gel could be formed by mixing DCA-Na solution with lysine and another anti-inflammatory drug, DSP (Tai Yu Chemical & Pharmaceutical Co., Ltd., Hsinchu County, China), and to determine the optimal pH value for gel formation, DCA-Na solution was mixed with L-lysine / DSP at different pH values ​​according to Table 9. The requirement was that the final concentration of DSP be 1 mg / mL or less.

[0083] [Table 9-1]

[0084] [Table 9-2]

[0085] [Table 10]

[0086] Figure 6 shows that when the L-lysine / DSP solution was added to the DCA-Na solution, all groups formed a clear solution. The pH values ​​of the mixed solutions ranged from 6.87 to 7.43 and 6.48 to 7.28 for the 5% and 1% DCA-Na solutions mixed with a 200 mg / mL L-lysine solution with a pH in the range of 4.0 to 7.0, and from 7.11 to 7.54 and 6.75 to 7.34 for the 5% and 1% DCA-Na solutions mixed with a 400 mg / mL L-lysine solution with a pH in the range of 4.0 to 7.0 (Table 10). In the 200 mg / mL L-lysine test, a mixture of 5% DCA-Na and L-lysine / DSP added to 0.9% physiological saline formed a gel in approximately 45 minutes at pH 4.0, while a mixture of 1% DCA-Na and L-lysine / DSP added to 0.9% physiological saline formed a gel in approximately 20 minutes at pH 4.0 (Figure 6, 6a). In the 400 mg / mL L-lysine test, a mixture of 5% DCA-Na and L-lysine / DSP added to 0.9% physiological saline formed a gel in approximately 30 minutes at pH 4.0 and 5.0, and in approximately 45 minutes at pH 6.0, while a mixture of 1% DCA-Na and L-lysine / DSP added to 0.9% physiological saline formed a gel in approximately 20 minutes at pH 4.0, and in approximately 30 minutes at pH 5.0 and 6.0 (Figure 6, 6b). This suggests that the DCA-Na solution can form a gel after being mixed with the L-lysine DSP solution, and that the time required for gel formation is reduced as the concentration of L-lysine increases. The preferred pH for the L-lysine / DSP solution is approximately pH 4.0–6.0.

[0087] In Example 5, when the final concentration of DCA-Na was 8.123 or 40.615 mg / mL, the final concentration of lysine was 46.154 or 92.308 mg / mL, and the final concentration of DSP was 0.999 mg / mL, the composition with 0.9% physiological saline added could form a gel. The composition with 0.9% physiological saline added could form a gel when the final pH of the composition was 6.48 to 7.38.

[0088] Example 6. Effects of DCA-Na and lysine containing DSP in porcine tissue. Three male pigs weighing at least 100 kg were anesthetized by intramuscular injection of 0.02 mg / kg atropine and inhalation of 3% isoflurane mixed with oxygen (O2) and 30-70% nitrous oxide (N2O). 0.5 mL of L-lysine / DSP solution (pH 6.0) was added to 1 mL of 1% or 5% DCA-Na solution and mixed until dissolved. The pigs were injected at different time points according to Table 11, with 0.9% saline, 1% or 5% DCA-Na solution, and L-lysine / DSP solution. Each injection site had an area of ​​9 cm², and the composition was injected to a depth of 0.5 cm at the center of each site. 54 injection sites were performed on each side of the pigs (total 108 sites / pig). On day 0, the animals were anesthetized by intramuscular injection of 0.02 mg / kg atropine and 6 mg / kg Zoletil 50. Adipose tissue samples were collected and cut in half down the middle. Photographs of the tissue sections were recorded and are shown in Figure 7.

[0089] [Table 11-1]

[0090] [Table 11-2]

[0091] Figure 7 shows that cell lysis occurred at the injection site when DCA-Na solution was injected at a shallow depth with lysine or lysine / DSP. Increasing the concentration or amount of DCA-Na resulted in a stronger cell lysis or inflammation, as evidenced by the observation of more widespread redness (Groups 2-5). The cell lysis or inflammation was considerably reduced, as evidenced by the reduced redness observed 7-14 days after injection. Furthermore, increasing the concentration of DSP reduced the degree and area of ​​redness at the injection site (Groups 6-9), suggesting that the addition of DSP, which has anti-inflammatory properties, can effectively reduce inflammation at the injection site.

[0092] Lysine compositions containing DCA-Na and DSP can effectively reduce fat while minimizing side effects such as inflammation and redness.

[0093] Example 7. Composition of DCA-Na and basic amino acids To test whether DCA-Na solution and other basic cationic amino acids form a gel after mixing, DCA-Na solution was mixed with acidic L-histidine (pH 5.0-5.2, Sigma-Aldrich) solution or L-arginine (pH 5.0-5.2, Sigma-Aldrich) solution, respectively, according to Tables 12 and 13.

[0094] Example 7.1. Composition of DCA-Na and L-histidine

[0095] [Table 12-1]

[0096] [Table 12-2]

[0097] [Table 12-3]

[0098] L-histidine solutions with concentrations higher than 2.86 mg / mL precipitated after being added to 1% DCA-Na solution (Figure 8, 8c-8f), and L-histidine solutions with concentrations higher than 11.43 mg / mL precipitated after being added to 5% DCA-Na solution (Figure 8, 8e-8f). Mixtures of DCA-Na and L-histidine with high concentrations of L-histidine began to form a gel in about 20 minutes when placed at 25°C, and mixtures placed at 37°C or 42°C formed a suspension (or precipitate) in a short time (Figure 8, 8b-8f). When 1% DCA-Na and L-histidine were added to 0.9% physiological saline, a gel formed in about 20 minutes when the L-histidine concentration was 1.43 mg / mL or higher (Figure 8, 8b-8e). When a mixture of 5% DCA-Na and L-histidine was added to 0.9% physiological saline, a gel formed in approximately 20 minutes when the L-histidine concentration was 2.86 mg / mL or higher (Figure 8, 8c-8e).

[0099] In Example 7.1, when the final concentration of DCA-Na was 7.54 to 48.00 mg / mL and the final concentration of L-histidine was 1.43 to 11.43 mg / mL, the composition to which 0.9% physiological saline was added can form a gel.

[0100] Example 7.2. Composition of DCA-Na and L-arginine

[0101] [Table 13]

[0102] Figure 9 shows that when 500 mg / mL of L-arginine solution was added to the DCA-Na solution, all groups formed a clear solution. However, only groups 1-10 formed a gel after being placed at 25°C and added to 0.9% physiological saline for approximately 60 minutes. All mixtures of DCA-Na and L-arginine placed at 37°C or 42°C did not form a gel at any of the test times.

[0103] In Example 7.2, when the final concentration of DCA-Na was 7.54 or 8.12 mg / mL and the final concentration of L-arginine was 115.38 or 142.86 mg / mL, the composition to which 0.9% physiological saline was added can form a gel.

[0104] These results revealed that although L-lysine, L-histidine, and L-arginine belong to the basic amino acid group, they require different concentrations to form a gel. For example, only high concentrations of L-arginine and low concentrations of DCA-Na formed a gel, and this took longer than with L-lysine and L-histidine. On the other hand, low concentrations of L-histidine were sufficient to form a gel. In terms of forming a gel composition with DCA-Na, lysine may be the most effective, followed by histidine, and arginine the least effective.

[0105] Example 8. Composition of DCA-Na and organic acid The inventors have shown that the pH value of a solution mixed with DCA-Na solution affects its gel-forming ability. To test whether DCA-Na solution and organic acid form a gel after mixing, DCA-Na solution was mixed with diluted acetic acid (Scharlau, Barcelona, ​​Spain) according to Table 14.

[0106] [Table 14-1]

[0107] [Table 14-2]

[0108] [Table 14-3]

[0109] 57.14 × 10 -3Acetic acid solutions with a concentration higher than % precipitated after being added to a 1% DCA-Na solution (8.80 mg / mL or less), and 45.45×10 -3 Acetic acid solutions with a concentration higher than % precipitated after being added to a 1% DCA-Na solution (9.60 mg / mL or more) (10b - 10f in Figure 10). 171.43×10 -3 Acetic acid solutions with a concentration higher than % precipitated after being added to a 5% DCA-Na solution (37.7 mg / mL or less), and 100.00×10 -3 Acetic acid solutions with a concentration higher than % precipitated after being added to a 5% DCA-Na solution (44.00 mg / mL or more) (10d - 10f in Figure 10). When the mixture of DCA-Na and acetic acid containing a higher concentration of acetic acid was placed at 25°C, it began to form a gel around 20 minutes. On the other hand, the mixtures placed at 37°C or 42°C formed a suspension (or precipitate) in a short time (10b - 10f in Figure 10). The mixture of 1% DCA-Na and acetic acid added to 0.9% physiological saline formed a gel around 20 minutes at an acetic acid concentration of 46.15×10 -3 % or higher (10b - 10e in Figure 10). The mixture of 5% DCA-Na and acetic acid added to 0.9% physiological saline formed a gel around 20 minutes when the acetic acid concentration was 92.30×10 -3 % or higher (10d - 10e in Figure 10).

[0110] In Example 8, when the final concentration of DCA-Na was 7.54 - 40.62 mg / mL and the final concentration of acetic acid was 46.15×10 -3 % - 142.86×10 -3 %, the composition added with 0.9% physiological saline could form a gel.

[0111] The present invention demonstrates that cell-lysing compounds, particularly deoxycholic acid or its DCA-Na salt, can form sustained-release gels, gel-forming solutions, or gel-forming suspensions after being mixed with low-pH amino acids (or cationic ions) or organic acids. Formulations of DCA-Na gels may include additional non-inflammatory agents, such as lysine aspirin or dexamethasone sodium phosphate, and the local anesthetic lidocaine, to reduce local inflammation. The present invention provides compositions of sustained-release cell-lysing compounds, such as deoxycholic acid or its salts, in gels or gel-forming solutions (or suspensions) for fat reduction, and with the addition of anti-inflammatory and / or local anesthetics for non-surgical reduction or removal of local fat, thereby reducing inflammation or other adverse effects, shortening the intervals between treatments, and shortening the overall treatment process. The injectable compositions of the present invention may optionally contain saline and may be in the form of a gel during or after injection.

Claims

1. An injectable composition for reducing fat, comprising a cell-lytic compound in a gel, gel-forming solution, or gel-forming suspension, The first component is the cell-lysating compound, where the cell-lysating compound is DCA-Na, where the concentration of DCA-Na is 7 to 51 mg / mL, and Pharmacologically acceptable excipients, and (i), (ii), and (iii): (i) L-lysine at a concentration of 11 to 145 mg / mL (ii) L-histidine at a concentration of 1.4 to 11.5 mg / mL, (iii) Concentration is 46-143 × 10 －３ % acetic acid, An injectable composition comprising a second component comprising at least one selected from the following.

2. The third component further contains an anti-inflammatory agent, where the anti-inflammatory agent is aspirin. The injectable composition according to claim 1, wherein the concentration of aspirin is 14 to 100 mg / mL.

3. The injectable composition according to claim 1, further comprising a local anesthetic as a fourth component, wherein the local anesthetic is lidocaine.

4. The injectable composition according to claim 2, wherein the anti-inflammatory drug is dexamethasone sodium phosphate (DSP).

5. An injectable composition according to any one of claims 1 to 4, for use in a method for reducing or removing local fat in a subject requiring reduction or removal of local fat, wherein the injectable composition is subcutaneously injected into the subcutaneous injection site of the subject.

6. The injectable composition according to claim 5, wherein the subcutaneous injection site is local fat in the face, chin, arm, waist, abdomen, or thigh of the subject.

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