WO2024168022A2

Movatterモバイル変換

Info

Publication number: WO2024168022A2
Application number: PCT/US2024/014780
Authority: WO
Inventors: John Downing; Bennet HARDING; Dileep JANAGAM
Original assignee: Tolmar Inc
Current assignee: Tolmar Inc
Priority date: 2023-02-10
Filing date: 2024-02-07
Publication date: 2024-08-15
Anticipated expiration: 2025-08-10
Also published as: WO2024168022A3

Abstract

Described herein are long-acting injectable compositions as well as methods of using such long-acting injectable compositions. The long-acting injectable compositions comprise degarelix (a decapeptide gonadotropin-releasing hormone (GnRH) receptor antagonist) or a pharmaceutically acceptable salt thereof. These compositions can also comprise a biodegradable polymer, and biocompatible organic solvent, optionally with one or more additives.

Description

Table 1 Description of Degarelix Citrate (DgC)-Drug Substance Powders

3. Degarelix mesylate preparation [00169] Degarelix mesylate was formed by dissolving the degarelix as degarelix acetate (purchased from the vendor) in water (~40 mg/mL concentration) and then adding the aqueous solution of strong acid methane sulfonic acid (1M) drop by drop. The molar ratio of degarelix free-base to methane sulfonic acid in the solution was varied as desired. The resulting solution was mixed for 10-15 mins and placed in the -80 °C freezer. The contents were lyophilized to remove the water (and freeze-drying may remove some amount of acetic acid). The dried powder was re-dissolved in water and freeze-dried again. The purity of the lyophilized degarelix mesylate powder was tested using HPLC analysis. 4. Preparation of Polymers [00170] Polymers were either purchased from Sigma Aldrich (RESOMER® polymers) or synthesized in-house. Poly(D.L-lactide-co-glycolide) (PLG) copolymers, poly-DL- lactide (PLA) polymers, or poly(D.L-lactide-co-caprolactone) (PDLCL or PLC) copolymers were produced using the following methods. The amounts of monomers (DL- lactide or glycolide or caprolactone) and initiator (e.g., glycolic acid, 1,6-Hexanediol, polyethylene glycol (PEG)) were selected to obtain a targeted initiator, monomer molar ratio, and weight average molecular weight for each investigated polymer. The monomer molar ratios and weight average molecular weights reported in individual examples are targeted values unless specified as actual or experimental. The polymerization vessel was lowered into a temperature-controlled oil bath. In the vessel, appropriate amounts of monomers (DL-lactide and/or glycolide and/or caprolactone, and initiator (glycolic acid or 1,6-Hexanediol) were added, the vessel contents were placed under a nitrogen atmosphere. The temperature of the vessel was increased until the reagents melted. A catalyst solution was made with appropriate amounts of stannous octoate and toluene and added to the vessel. The vessel was then heated to about 140-170°C under a nitrogen atmosphere for about 4-18 hours (depending on the polymer of interest) with constant stirring. Then, the vessel was evacuated to remove unreacted monomers, and the monomers were vacuum- distilled out of the polymerization mixture. The hot melt was then extruded into cooling pans. After cooling, the solid mass was broken up into smaller pieces. The polymer was purified as needed using the solvent/non-solvent-induced phase separation method. In the notation, PLC-H-P or PLA-H-P or PLG-H-P used here ‘H’ denotes if the polymer is acid terminated and ‘P’ denotes additional polymer purification step by solvent/non-solvent induced phase separation method. 5. Preparation of bulk polymer solutions and prefilled syringes (Syringe A) of polymer solution [00171] Polymer bulk solutions were prepared by weighing a known amount of each polymer material into individual Flack-Tek jars of the desired size. A known amount of NMP was added to each polymer, blanketed with nitrogen and the containers were placed on a horizontal jar mill or Turbula for mixing at room temperature. Jars were mixed until a visually clear homogenous polymer solution is obtained indicating the complete dissolution of the polymer in the solvent. Prefilled syringes (Syringe A) containing polymer delivery system were prepared by weighing the required amounts of polymer solution into 1.2 mL female polypropylene syringe barrel with Luer lock and plunger tip and capped with male polypropylene syringe cap. Filled syringes were then packaged in labeled foil pouches with a desiccant pack and the pouches were sealed. After filling the syringes with the formulations, the filled syringes were stored under refrigerated conditions (e.g., 2-8 ºC) or accelerated conditions (≥25°C see individual experiments). The syringes may be irradiated via e-beam irradiation (see individual experiment). 6. Preparation of DgA-DS or DgC-DS/organic-solvent bulk solution and prefilled syringes [00172] To produce the drug/solvent bulk solution comprising the API (DgA or DgC), the desired amount of the DgA-DS or DgC-DS was combined with the solvent NMP or DMSO or combination of solvents with or without a co-solvent or other additives, in the indicated amounts (see individual experiments below). The API and solvent were combined in a glass vial or jar and blanketed with nitrogen. Jars were mixed using the jar mill, Turbula, or shaker at room temperature until homogeneous. DgA or DgC are very soluble in NMP or DMSO and can load higher concentrations (at least 40% w/w). After dissolving the API in the organic solvent, the bulk solution was manually filled into syringes and capped with a tip cap. The syringes used for filling were either 1.2 mL male polypropylene syringes or 1 mL male cyclic olefin copolymer syringes. Filled syringes were then packaged in labeled foil pouches with a desiccant pack and the pouches were sealed. After filling the syringes with the formulations, the filled syringes were stored under refrigerated conditions (e.g., 2- 8 ºC) or accelerated conditions (≥25°C see individual experiments). The syringes may be irradiated via e-beam or gamma irradiation (see individual experiment) for terminal sterilization. 7. Preparation of Reconstituted Formulation for testing: [00173] Immediately before testing, “A” and “B” syringes were coupled and mixed by cycling the contents from one syringe to the other for 30-45 cycles. The mixed formulation was finally transferred to the male dosing syringe for delivery and testing. A safety needle of 20 g was used for delivering the formulation when needed. 8. Preparation methods for Single-Syringe Formulation [00174] Selected degarelix (acetate or citrate form) was dissolved in organic solvent (Ex: NMP or DMSO) at desired amounts and then mixed with the desired amount of liquid polymer solution (see section 5 for bulk polymer solution preparation) in a scintillation vial, and the vial was put on a horizontal roller mixer to roll for 24-48 h for mixing. After mixing, the single-syringe formulations (solution/suspension) were considered ready for testing. [00175] As an alternative to the above method of preparation, selected degarelix (acetate or citrate form) and liquid polymer solution (see section 7 for bulk polymer solution preparation) were weighed into a scintillation vial, and the vial was put on a horizontal roller mixer to roll for 24-48 h for mixing. After mixing, the single-syringe formulations (solution/suspension) were considered ready for testing. 9. Preparation of Single-Syringe Formulation with added stabilizers [00176] The stabilizer excipient can be selected from a list of acids which include polycarboxylic acids or their salts (Ex. citric acid), strong acids (Ex. methanesulfonic), hydrophobic acids or their salts (Ex. pamoic acid, palmitic acid, lauric acid, sodium dodecyl sulfate). Degarelix acetate powder, a stabilizer excipient(s) (anhydrous citric acid), and organic solvent (e.g., NMP or DMSO) were weighed into a scintillation vial, and the vial was put on a horizontal roller mixer to roll for 24-48 h for mixing. After mixing, desired amounts of polymer bulk solution (see section 5) were added to the drug solution vial, and the vial was put on a horizontal roller mixer and continued to mix for 24-48 h. After mixing, the single-syringe formulations were considered ready for testing. [00177] As an alternative preparation method to the above, desired amounts of DgA-DS, a stabilizer excipient(s) (e.g., anhydrous citric acid), and liquid polymer solution (see section 5 for bulk polymer solution preparation) were weighed into a scintillation vial, and the vial was put on a horizontal roller mixer to roll for 24-48 h for mixing. After mixing, the single- syringe formulations (solution/suspension) were considered ready for testing. 10. XRD Characterization of the DgA-DS solutions [00178] Samples were prepared and analysis was performed at ambient temperature using the Rigaku MiniFlex XRD instrument according to standard protocols. After a sequence of samples was run, the data was processed using PDXL2 software. Degarelix acetate did not show any crystallinity as an API powder, or at a concentration of 66 mg/mL in water. At higher concentrations, the peptide forms a gel. The gel was not crystalline. In theory, the gel exhibits peaks indicating a higher order structure attributed to the stacking of beta-sheets forming aggregates resulting in the gel formation. The addition of salts can aid the gelation to form at a lower concentration than without the salts. No gelation occurred in the presence of NMP solvent even when water was present. 11. Analysis of degarelix (assay) and Total related compounds in the formulations [00179] Degarelix/organic solvent solutions were prepared for HPLC analysis by dilution to volume with mobile phase A (0.1% trifluoroacetic acid water) and mixing thoroughly by vortex. Dilutions were performed as needed using volumetric glassware and mixed thoroughly by vortex mixing. A second dilution of 2 mL to 20 mL was performed with mobile phase A to obtain a working sample. [00180] For the HPLC test sample preparation of reconstituted/single-syringe formulations that contain polymer, syringe A and syringe B were coupled together and mixed for 45 cycles. The reconstituted product was dispensed from the final mixed single- syringe formulation content into a 50 mL volumetric flask and the weight was recorded. 2.0 mL of mobile phase B (0.1% trifluoroacetic acid acetonitrile) was added and mixed by swirling. 3.0 mL of mobile phase A was added and mixed by swirling. Samples were placed in a water bath at 55 ºC ± 2 ºC for 30 minutes and then cooled to room temperature. Samples were diluted to volume with mobile phase A and mixed thoroughly by vortex, followed by dilution to 20 mL with mobile phase A with thorough mixing. A second dilution of 2 mL to 20 mL was performed with mobile phase A to obtain a working sample. The sample solutions were filtered through a 0.45 um PTFE syringe filter into amber HPLC vials before collection into the HPLC vial. [00181] HPLC analysis for assay and related compounds was performed using an Agilent AdvanceBio Peptide 3.0 x 100 mm, 2.7 um column and an Agilent AdvanceBio Peptide Map Guard 3.0 x 5mm 2.7 um guard column at 30°C with a flow rate of 0.75 mL/min. The runtime was 15 minutes with a 10-µL injection for assay and related compounds for the solution gel depot formulation, and a 20-µL injection for assay and related compounds for the polymer depot formulation. The mobile phases were 0.1% trifluoroacetic acid water for mobile phase A and 0.1% trifluoroacetic acid acetonitrile for mobile phase B. Detection was performed with a diode array detector set at 220 nm. The method used a gradient. See Table 2 below for the gradient method for an example chromatogram.

12. Recovery of drug from different Degarelix/Organic solvent formulations [00182] Prefilled syringes of degarelix/organic solvent formulations were prepared as described in section 6. Recovery of drugs from different formulations was tested using the HPLC method (see section 11) and is listed in Table 3.

Table 3 Drug recovery testing from prefilled syringes of organic-solvent based formulations

(“F#” denotes “Formulation Number”) [00183] Table 3 Observations: All the formulations showed good recoveries (90.0- 110.0%), indicating the applicability of the method and the initial stability of the API in different solvent systems. 13. Stability of degarelix in organic solvents (Syringe B) [00184] Prefilled syringes of DgA-DS/NMP or DgA-DS/DMSO solutions were prepared as described in section 6. The samples were not terminally sterilized. The composition of the stability samples is shown in Table 4. The sealed foil pouches containing prefilled syringes were paced at accelerated storage conditions (25±2°C) for 6 months and tested for drug assay or recovery and related compounds (see section 11 for procedure) at each time pull (0, 1, 3 and 6 months). The results of this study are presented in Figure 1. No degradation of the drug was observed at 6 months, and a slight increase in percentage recoveries over time could be attributed to the solvent loss/absorption in prefilled syringes at accelerated conditions. Table 4 Compositions Of Formulations

[00185] As shown in Figure 1 degarelix acetate was found to be stable in NMP and DMSO (at studied concentrations) at 25°C for 6 months (accelerated conditions). There was no significant increase in the related compounds. At 6 months, the total related compounds were below 2%. The observed slight increase in percentage recoveries over time for the F1, F2, and F5 formulations on storage could be attributed to the solvent loss over time at accelerated storage conditions from the prefilled syringes or stopper absorption. No gelation was observed in the formulations F1, F2, and F5. F5 formulation was made at a higher concentration than F2, and both were found to be stable and did not show any gelation. This supports that higher concentration, such as 40% w/w, is also feasible. 14. Terminal sterilization feasibility of degarelix solutions via e-beam or Gamma irradiation. [00186] The prefilled syringes of degarelix/organic solvent were exposed to e-beam or gamma radiation at different doses and tested the drug concentrations or recoveries for pre and post-irradiation samples using HPLC (see section 11) and the results are presented in Table 5.

Table 5 Results Of Irradiation Feasibility Studies

(“F#” denotes “Formulation Number”) [00187] Table 5 Observations: These results showed that depending on the irradiation type, irradiation dose, concentration of the drug, salt form, and stabilizer, the degarelix solutions in an organic solvent are feasible for terminal sterilization by irradiation. [00188] F4-e25-2 formulation of degarelix citrate showed better stability upon e-beam irradiation compared to degarelix acetate formulation F1-e25-2. [00189] In general, 35% w/w drug solution samples (F1-e34, F2-e34, F1-g22) showed better stability upon irradiation compared to 40% w/w drug solution formulations (F6-e34, 5-e34, F6-g22). [00190] Formulations F1-e25, F2-e25, F3-e25, and F4-e25 were tested for total related compounds, no significant increase was observed post-irradiation of the samples via e- beam. 15. Stability of Irradiated samples in accelerated storage conditions [00191] Irradiated prefilled syringes of degarelix/organic solvent formulations were stored at different accelerated storage conditions (25°C) and tested for assay at 30 and 120 days using the HPLC method (see section 11). Results are presented in Table 6. Table 6 Accelerated Storage Stability Testing Of Irradiated Prefilled Syringes Of Degarelix Solution Formulations

[00192] Table 6 Observations: No pronounced degradation of the degarelix was observed in the irradiated samples over the studied period and a slight increase in percentage recoveries over time could be attributed to the solvent loss or absorption in prefilled syringes at accelerated storage conditions. 16. Compatibility of degarelix acetate with a Polymer solution or monomer solution [00193] A known amount of DgA-DS was combined with known amounts of selected polymer solutions or monomer solution and incubated at 50 °C and/or at room temperature, and tested for drug recovery at selected time points. See Table 7 and Table 8 for details. Table 7 Degarelix Acetate Polymer Or Monomer Solution Compatibility Testing

(“F#” denotes “Formulation Number”). NA indicates: Not Available Table 8 Degarelix Acetate Polymer Or Monomer Solution Compatibility Testing

(“F#” denotes “Formulation Number”) NA indicates: Not Available [00194] Table 7 Observations: F6 sample was a control sample that does not contain any polymer or monomers and was included in the study as a control to understand if the solvent causes any degradation. The F6 sample showed minimal signs of degradation (1% drop in assay after 24 h at 50°C), indicating that the drug is stable in NMP, and any degradation observed in other samples could be attributed to the presence of polymers/monomers. Bold text in Table 7 for Samples F8, F10 and F15 indicates that these samples exhibited the most promising drug stability (e.g., less degradation of drug over the tested time period). [00195] F16 and F17 are samples where DgA-DS was mixed with monomer solutions (Lactide/NMP or Glycolide/NMP), and it was observed that almost 90-100% of the drug was degraded by 7 days at room temperature, and 86-99% in 24 h at 50°C. These results indicated that the presence of monomers could induce significant degradation of the degarelix. These results indicate that purification of polymers to remove the residual or unreacted monomers will improve the stability of the API. [00196] Among F8-F14 samples that contain unpurified polymers, glycolic acid (GA) terminated PLC containing samples F8 and F10 have shown better compatibility with the drug compared to the PLG or PLC-PEG polymers. F8 and F10 samples showed ≤5% (after 24 h at 50°C) and ≤1.2% (after 7 days at room temperature) degradation, whereas other samples showed significantly higher degradation of the degarelix. [00197] F15 sample that contains purified acid-terminated PLG polymer also showed promise, 7.2% (after 24 h at 50°C) and 4.5% (after 7 days at room temperature) degradation. Although there are other samples such as F9, F11, and F14 that contain similar polymers (acid-terminated PLGs) with a different monomer composition, the higher stability of degarelix in F15 could be related to the use of purified polymer in F15 compared to the unpurified versions in F9, F11, and F14. As discussed earlier, the presence of residual monomers could be detrimental to the API stability, so purification of the polymer helped in improving the stability. [00198] Acid-terminated PLC polymer showed improved stability of degarelix compared to other polymers, even without purification. Based on these results, purified acid- terminated PLC polymer showed promising potential for use in formulating a single syringe formulation. [00199] Table 8 Observations: Bold text in Table 8 for Samples F20, F24, F28 and F32 indicates that these samples exhibited the most promising drug stability (e.g., less degradation of drug over the tested time period). Based on the results from Table 8, if the stability trends of degarelix in polymer solutions are to be ranked with respect to the polymer composition, the rank was PLC>PLA>PLG, and acid-terminated>ester-terminated polymer. These results confirmed earlier observations, that purified acid-terminated PLC polymers are more compatible with degarelix and showed promising potential as a polymer of choice for a single syringe formulation. In addition to purified acid-terminated PLC, purified acid- terminated PLA polymer also showed promising stability for a single syringe formulation. [00200] Degradation of DgA-DS was observed in presence of polymer solution. Therefore, additional studies were conducted where modified degarelix (DgC-DS instead of DgA-DS) or the use of stabilizer excipient were evaluated to further improve the stability of the degarelix with a goal of making a stable single syringe formulation. 17. LC-MS analysis of the single syringe formulations [00201] The working samples in the volumetric flask were obtained from HPLC sample preparation followed by filtration through a 0.45 µm syringe filter into an amber HPLC vial. The liquid chromatography analysis was performed using Agilent AdvanceBio Peptide 3.0 x 100 mm, 2.7 um column and an Agilent AdvanceBio Peptide Map Guard 3.0 x 5mm 2.7 um guard column at 30 °C. The flow rate was set at 0.35mL/min with 20µL injection and the gradient is listed below in Table 9. Mobile phase A was 0.1% trifluoroacetic acid water and mobile phase B was 0.1% trifluoroacetic acid acetonitrile. Both UV (at 220nm) and mass spec signals are collected. Table 9 Gradient Method For LC Run

[00202] The mass spec signals were collected using ESI resource at positive mode. The parameter settings of the ESI source are listed below in Table 10. Table 10 Esi Source Setting Of Mass Spectrometer

18. Improved stability of degarelix using DgC-DS or Citric acid stabilizer excipients [00203] Single-syringe formulations of DgA-DS and DgC-DS (different moles, see section 2 Table 1) were prepared as per the procedure in section 8. Single-syringe formulations of DgA-DS with added citric acid excipient were prepared as per the procedure in section 9. Single-syringe formulations of DgA-DS with and without stabilizer (different moles of anhydrous citric acid), and DgC-DS were tested for stability under accelerated storage conditions at 50°C using LC-MS. The m/z values of the molecular ion for the main and related compound peaks are summarized in Table 11. Table 11 Degradation And Impurities Of Single Syringe Formulations

(“F#” denotes “Formulation Number”) [00204] Table 11 Observations: This accelerated stability at 50°C evaluated the effect of i) citric acid excipient, and ii) degarelix citrate form over degarelix acetate form on the stability of degarelix in the polymer-based single-syringe formulations. [00205] If the total impurities (area sum%) of F37 vs. (F34, F35, F36, F45, F46, F47, F48), and F41 vs. (F38, F39, F40) is compared, DgA-DS containing formulations without citric acid F37 (15.0% impurities) and F41(15.2% impurities) generated significantly higher amount of impurities after only 14 days compared to the formulations that contained DgC- DS (<5% impurities after 21 days) or citric acid as an excipient (<8% after 18 days). Lower levels of acylated impurities were observed for citric acid-based formulations than formulations that do not contain any citric acid. [00206] Addition of citric acid as a stabilizer excipient to the drug-polymer formulation or using it as part of the alternate salt form that is prepared by co-lyophilization of DgA-DS with citric acid, both resulted in a similar improvement of drug stability on accelerated storage, and the lower impurity levels were observed. Further, this preliminary data suggested that stabilizers such as citric acid can be just added as an excipient to the formulation to improve the drug stability in presence of polymer, and the additional process of lyophilization to make DgC-DS form may be eliminated to simplify the manufacturing process. [00207] Citric acid was added as an excipient to the formulation at four different molar levels [0.5 (F48), 1 (F45), 2 (F46), and 3 (F47) moles to 1 mole of degarelix). [00208] F48 with 0.5 mol citric acid showed 7.8 % total impurities at day 18 vs. F37 with no citric acid showed 15.0% total impurities at day 14. F45 with 1 mol of citric acid showed 3.6% total impurities at day 18. Although 0.5 moles of citric acid helped in improving the stability of DgA-DS compared to no additive version, higher levels of citric acid showed much better improvement in drug stability. This set of results indicates that higher than 0.5 moles of citric acid to degarelix is needed to achieve better drug stability. The addition of 1:1 or molar excess of citric acid to the degarelix showed promising results. 19. Animal dosing [00209] Prefilled syringes of liquid polymer solutions (Syringe A) and degarelix/organic solvent solutions (Syringe B) were prepared as described in section 5 and section 6. The samples were not e-beamed. Degarelix release rates of these formulations were obtained using a rat model. Male rats were each injected with a single subcutaneous injection of the reconstituted formulations obtained by mixing Syringe A and Syringe B versions of different formulations. The composition of the formulations and the dosing details are listed in Table 12. [00210] At predetermined time points, rats were bled and plasma degarelix levels were determined using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Each data point is based on an average plasma degarelix concentration. Six rats were dosed for each group, with sparse blood sampling performed for early time points. FIRMAGON^® (degarelix for injection) was included in the study as a control. The control was injected on day 0 and Day 28 at 15 mg/kg to understand steady state and accumulation, whereas test groups received a single injection of 45 mg/kg. A 120 mg vial of FIRMAGON^® was reconstituted by adding 3 mL of sterile water for injection provided with the FIRMAGON^® kit, and the resulting reconstituted solution was at 40 mg/mL concentration of degarelix. Table 12 Non-Clinical Study Dosing Details

(“F#” denotes “Formulation Number”) [00211] In vivo degarelix release in rats for polymer-based formulations of degarelix at 28-days post injection is presented in Figure 2. Figure 3 shows a magnified version of day- 0 to day-3 levels to show the Cmax levels. As seen in Figures 2 and 3, all the polymer-based formulations (Groups F-TA5 to L-TA11) showed higher initial release (Cmax) than control (Group A-CA1) which could be attributed to the higher injection dose. Groups F-TA5 to L- TA11 were injected at 3 times higher doses (45 mg/kg) than control A-CA1 (15 mg/kg). [00212] Figures 2 and 3 show that, although polymer-based formulations were injected with a 3 times higher dose than the control, the initial Cmax numbers observed for polymer- based test articles were much less than the 3*Cmax of the control, which indicates that the polymer-based formulations may have resulted in a better encapsulation of the drug and lower release of the drug initially. [00213] In vivo degarelix release in rats for polymer-based formulations of degarelix at 63-days post injection is presented in Figure 4. Figure 5 shows a magnified version of the same data in Figure 4 to better illustrate the release of the different formulations. Figures 6 and 7 show that all of the formulations released drug that resulted in measurable levels of degarelix in vivo for greater than 60 days. [00214] In vivo degarelix release in rats for polymer-based formulations of degarelix at 119-days post injection is presented in Figure 6. Figure 7 shows a magnified version of the same data in Figure 6 to better illustrate the release of the different formulations. Figures 6 and 7 show that all of the formulations released drug that resulted in measurable levels of degarelix in vivo for greater than 90 days (at least 119 days). Also, the formulation L-TA11 (delarelix citrate) showed notably higher levels of drug in vivo compared to other formulations throughout the testing period. [00215] In vivo testosterone concentration levels (ng/mL) in rats for the polymer-based formulations of degarelix API in this example at 105-days post injection is presented in Figure 8. Together, the results show that all of the formulations had plasma concentration of greater than 1 ng/mL which is the minimum drug concentration to induce chemical castration. In addition, the testosterone level remained below the “historical testosterone baseline” level and testosterone suppression occurred by day 4. Lastly, a second dose of FIRMAGON^® (A-CA1) at Day 28 did not suppress testosterone concentration levels any more than the other formulations dosed once on Day 0. [00216] Table 13 shows the PK data for the experiment through Day 119 post-injection described above. The Cmax for all polymer-based formulations occurred earlier based on Tmax compared to Firmagon; this could lead to a faster reduction in testosterone levels. Table 13 PK Data

*Dosed once on Day 0 and once on Day 28. Dose normalized parameters are calculated off total dose. [00217] Various modifications of the above-described invention will be evident to those skilled in the art. It is intended that such modifications are included within the scope of the following claims.

Claims

CLAIMS 1. A composition, comprising: a therapeutically effective amount of degarelix or a pharmaceutically acceptable salt thereof; a biodegradable polymer; and a biocompatible solvent, wherein: the composition is formulated for subcutaneous injection into a subject; a single dose of the composition is about 2.5 mL or less or about 2.0 mL or less; and upon injection into the subject, the composition forms an in situ depot that releases the degarelix or pharmaceutically acceptable salt thereof over a time period of from about 1 month to about 6 months.

2. The composition of claim 1, wherein the in situ depot releases the degarelix or pharmaceutically acceptable salt thereof over a time period selected from the group consisting of: at least about 1 month, at least about 1.5 months, at least about 2 months, at least about 2.5 months, at least about 3 months, at least about 3.5 months, at least about 4 months, at least about 4.5 months, and at least about 5 months.

3. The composition of claim 1, wherein the pharmaceutically acceptable salt of the degarelix is selected from degarelix acetate, degarelix citrate, degarelix pamoate, degarelix palmitate, and degarelix mesylate.

4. The composition of claim 1, wherein the biodegradable polymer comprises monomer residues selected from the group consisting of lactide, glycolide, caprolactone, p- dioxanone, trimethylene carbonate, thylene oxide, propylene oxide, sebacic anhydride, diketene acetals/diols, and combinations thereof.

5. The composition of claim 4, wherein the biodegradable polymer comprises lactide and glycolide monomer residues.

6. The composition of claim 5, wherein a molar ratio of the lactide to glycolide monomer residues is from about 45:55 to about 99:1.

7. The composition of claim 7, wherein a molar ratio of the lactide to glycolide monomer residues is from about 50:50 to about 90:10.

8. The composition of 4, wherein the biodegradable polymer comprises: lactide monomer residues and monomer residues selected from the group consisting of ε- caprolactone, trimethylene carbonate, and combinations thereof.

9. The composition of claim 8, wherein a molar ratio of the lactide monomer residues to the ε-caprolactone and/or trimethylene carbonate monomer residues is from about 10:90 to about 90:10.

10. The composition of claim 9, wherein a molar ratio of the lactide monomer residues to the ε-caprolactone and/or trimethylene carbonate monomer residues is from about 25:75 to about 75:25.

11. The composition of claim 10, wherein a molar ratio of the lactide monomer residues to the ε-caprolactone and/or trimethylene carbonate monomer residues is about 75:25.

12. The composition of any one of claims above, wherein the biodegradable polymer comprises at least one carboxylic acid end group, at least one hydroxyl end group, or at least one hydroxy end group and is substantially free of terminal carboxy end groups.

13. The composition of any one of claims 8-11, wherein the biodegradable polymer is poly(D,L-lactide-co-glycolide) (PLG), poly(D,L-lactide) (PLA), or poly(D,L-lactide-co-ε- caprolactone) (PLC).

14. The composition of any one of claims 8-11, wherein the biodegradable polymer is poly(D,L-lactide-co-ε-caprolactone) (PLC).

15. The composition of claim 14, wherein the PLC comprises at least one carboxylic acid end group.

16. The composition of claim 15, wherein the PLC is a carboxylic acid-initiated PLC.

17. The composition of claim 16, wherein the carboxylic acid is glycolic acid or lactic acid.

18. The composition of claim 14, wherein the biodegradable polymer is acid-initiated 75:25 PLC.

19. The composition of any one of claims 5-7, wherein the biodegradable polymer is PLG.

20. The composition of claim 19, wherein the PLG comprises at least one hydroxyl end group.

21. The composition of 20, wherein the PLG is a core-diol-initiated PLG.

22. The composition of claim 21, wherein the core diol is 1,6-hexane-diol.

23. The composition of claim 19, wherein the PLG is a mono functional alcohol-initiated PLG.

24. The composition of claim 23, wherein the alcohol is dodecanol.

25. The composition of claim 19, wherein the biodegradable polymer is acid-initiated 50:50 PLG.

26. The composition of claim 22, wherein the biodegradable polymer is 1,6-hexane-diol- initiated 75:25 PLG.

27. The composition of claim 22, wherein the biodegradable polymer is 1,6-hexane-diol- initiated 85:15 PLG.

28. The composition of claim 13 wherein the biodegradable polymer is dodecanol- initiated PLA.

29. The composition of claim 1, wherein the biocompatible polymer has a weight average molecular weight of from about 5 kDa to about 60 kDa.

30. The composition of claim 1, wherein the biocompatible polymer has a weight average molecular weight of from about 5 kDa to about 35 kDa.

31. The composition of claim 1, wherein the biocompatible polymer comprises less than about 0.5 wt%, or less than about 0.4 wt%, or less than about 0.3 wt%, or less than about 0.2 wt% or less than about 0.1 wt% unreacted lactide monomer.

32. The composition of claim 1, wherein the amount of biodegradable polymer in the composition is from about 10% to about 60% by weight of the composition.

33. The composition of claim 1, wherein the amount of biocompatible solvent in the composition is from about 10% to about 85% by weight of the composition.

34. The composition of claim 1, wherein the therapeutically effective amount of degarelix or pharmaceutically acceptable salt in the composition is from about 10% to about 40% by weight of the composition.

35. The composition of claim 1, wherein the therapeutically effective amount of degarelix or pharmaceutically acceptable salt thereof in the composition is from about 8 wt% to about 39 wt% degarelix free base equivalent.

36. The composition of claim 1, wherein the therapeutically effective amount of degarelix or pharmaceutically acceptable salt thereof is from about 40 mg – 500 mg.

37. The composition of claim 1, wherein the therapeutically effective amount of degarelix or pharmaceutically acceptable salt thereof is from about 80 mg – 500 mg.

38. The composition of claim 1, wherein the therapeutically effective amount of degarelix or pharmaceutically acceptable salt thereof is from about 120 mg – 500 mg.

39. The composition of claim 1, wherein the biocompatible solvent is selected from the group consisting of: N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO), dimethylacetamide (DMA), benzyl benzoate (BnBzO), polyethylene glycol 15 hydroxystearate, methyl ethyl ketone, methyl lactate, benzyl alcohol, propylene carbonate (PC), triacetin, tributyl citrate, acetyl tributyl citrate, acetyl triethyl citrate, triethyl citrate, diethylene glycol monomethyl ether, ethyl acetate, N-ethyl-2-pyrrolidone, glycofurol and combinations thereof.

40. The composition of claim 39, wherein the biocompatible solvent is NMP or DMSO.

41. The composition of claim 1, further comprising one or more additives selected from the group consisting of polysorbate 20, polysorbate 80, poloxamer 188, sorbitan trioleate, lecithin (e.g., soya or egg), polyethylene glycol (PEG), PEG 300, 2-pyrrolidone, alpha- tocopherol, Vitamin E TPGS, sucrose cocoate, sucrose stearate, sucrose laurate, proline, arginine, sodium metabisulfite butylated hydroxyanisole, butylated hydroxyquinone, butylhydroxyanisol, hydroxycoumarin, butylated hydroxytoluene, cephalm, ethyl gallate, propyl gallate, octyl gallate, lauryl gallate, propyl-hydroxybenzoate, trihydroxybutylrophenone, vitamin E, lecithin, ethanolamine, ZnCl2, MgCl2, CaCl2, DL- methioninecitric acid, dimethylphenol, dibutylphenol, ethylenediaminotetraacetic acid (EDTA), ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), ascorbic acid, , nitrilotriacetic acid, n-hydroxyethylethylenediaminetriacetic acid (HEDTA), mercaptoethanol, and combinations thereof.

42. The composition of claim 1, wherein a single dose of the composition is about 2.5 mL or less, about 2.4 mL or less, about 2.3 mL or less, about 2.2 mL or less, about 2.1 mL or less, about 2.0 mL or less, about 1.9 mL or less, about 1.8 mL or less, about 1.7 mL or less, about 1.6 mL or less, about 1.5 mL or less, about 1.4 mL or less, about 1.3 mL or less, about 1.2 mL or less, about 1.1 mL or less, about 1 mL or less, about 0.75 mL or less, about 0.5 mL or less, or about 0.375 mL or less.

43. The composition of claim 1, wherein the degarelix or pharmaceutically acceptable salt thereof is lyophilized, and wherein the biodegradable polymer is dissolved in the biocompatible solvent.

44. The composition of claim 1, wherein the degarelix or pharmaceutically acceptable salt thereof is dissolved in a first biocompatible solvent, and wherein the biodegradable polymer is dissolved in a second biocompatible solvent.

45. The composition of claim 44, wherein the first and second biocompatible solvent are the same.

46. The composition of any one of claims 1-45, wherein the composition has been terminally sterilized or sterile filtered.

47. The composition of claim 46, wherein the composition has been terminally sterilized by e-beam.

48. A pharmaceutical composition, comprising: (a) about 10-30 wt% of degarelix acetate; (b) about 20-80 wt% of N-methyl-2-pyrrolidone (NMP); and (c) about 10-50 wt% of 75:25 poly(lactide-co-ε-caprolactone) (PLC) copolymer having at least one carboxylic acid end group and having a weight average molecular weight of about 5-30 kDa.

49. The pharmaceutical composition of claim 48, wherein the composition comprises: (a) about 16 wt% of degarelix acetate; (b) about 57 wt% of N-methyl-2-pyrrolidone (NMP); and (c) about 27 wt% of 75:25 poly(lactide-co-ε-caprolactone) (PLC) copolymer having at least one carboxylic acid end group and having a weight average molecular weight of about 17 kDa.

50. The pharmaceutical composition of claim 48, wherein the composition comprises: (a) about 16 wt% of degarelix acetate; (b) about 57 wt% of NMP; and (c) about 27 wt% of 75:25 PLC copolymer having at least one carboxylic acid end group and having a weight average molecular weight of about 8 kDa.

51. A pharmaceutical composition, comprising: (a) a first container comprising 30-70 wt% of 75:25 PLC copolymer having at least one carboxylic acid end group and having a weight average molecular weight of 5-30 kDa, and 30-70 wt% of N-methyl-2- pyrrolidone (NMP); and (b) a second container comprising 20-40 wt% of degarelix acetate and 60-80 wt% NMP.

52. The pharmaceutical composition of claim 51, wherein: (a) the first container comprises 50 wt% of 75:25 PLC copolymer having at least one carboxylic acid end group and having a weight average molecular weight of 17 kDa, and 50 wt% of N-methyl-2-pyrrolidone (NMP); and (b) the second container comprises 35 wt% of degarelix acetate and 65 wt% NMP.

53. The pharmaceutical composition of claim 51, wherein: (a) the first container comprises 70 wt% of 75:25 PLC copolymer having at least one carboxylic acid end group and having a weight average molecular weight of 8 kDa, and 30 wt% of NMP; and (b) the second container comprising 35 wt% of degarelix acetate and 65 wt% NMP.

54. The pharmaceutical composition of any one of claims 48-53, wherein unreacted lactide and caprolactone monomers in the PLC copolymer are less than about 1.0 wt%.

55. The pharmaceutical composition of any one of claims 48-53, wherein unreacted lactide and caprolactone monomers in the PLC copolymer are less than about 0.5 wt%, than about 0.4 wt %, less than about 0.3 wt %, less than about 0.2 wt% and less than about 0.1 wt%.

56. A pharmaceutical composition, comprising: (a) about 10-30 wt% of degarelix acetate; (b) about 20-80 wt% of NMP; and (c) about 10-50 wt% of poly(D,L-lactide) (PLA) polymer having at least one carboxylic acid end group and having a weight average molecular weight of about 5-30 kDa.

57. The pharmaceutical composition of claim 56, wherein the composition comprises: (a) about 16 wt% of degarelix acetate; (b) about 57 wt% of NMP; and (c) about 27 wt% of poly(D,L-lactide) (PLA) polymer having at least one carboxylic acid end group and having a weight average molecular weight of about 23 kDa.

58. A pharmaceutical composition, comprising: (a) a first container comprising 30-70 wt% of PLA polymer having at least one carboxylic acid end group and having a weight average molecular weight of 5-30 kDa, and 30-70 wt% of NMP; and (b) a second container comprising 20-40 wt% of degarelix acetate and 60-80 wt% NMP.

59. The pharmaceutical composition of claim 58, wherein: (a) the first container comprising 50 wt% of PLA polymer having at least one carboxylic acid end group and having a weight average molecular weight of 23 kDa, and 50 wt% of NMP; and (b) the second container comprising 35 wt% of degarelix acetate and 65 wt% NMP.

60. The pharmaceutical composition of any one of claims 56-59, wherein unreacted lactide monomers in the PLA polymer are less than about 1.0 wt%.

61. The pharmaceutical composition of any one of claims 56-59, wherein unreacted lactide monomers in the PLA polymer are less than about 0.5 wt%, than about 0.4 wt %, less than about 0.3 wt %, less than about 0.2 wt% and less than about 0.1 wt%.

62. A pharmaceutical composition, comprising: (a) about 10-30 wt% of degarelix acetate; (b) about 20-80 wt% of NMP; and (c) about 10-50 wt% of 75:25 poly(D,L-lactide-co-glycolide) (PLG) copolymer having at least one carboxylic acid end group and having a weight average molecular weight of about 5-30 kDa.

63. The pharmaceutical composition of claim 62, wherein the composition comprises: (a) about 16 wt% of degarelix acetate; (b) about 57 wt% of NMP; and (c) about 27 wt% of 75:25 poly(D,L-lactide-co-glycolide) (PLG) copolymer having at least one carboxylic acid end group and having a weight average molecular weight of about 17 kDa.

64. A pharmaceutical composition, comprising: (a) a first container comprising 30-70 wt% of 75:25 PLG copolymer having at least one carboxylic acid end group and having a weight average molecular weight of 5-30 kDa, and 30-70 wt% of NMP; and (b) a second container comprising 20-40 wt% of degarelix acetate and 60-80 wt% NMP.

65. The pharmaceutical composition of claim 64 wherein: (a) the first container comprises 50 wt% of 75:25 PLG copolymer having at least one carboxylic acid end group and having a weight average molecular weight of 17 kDa, and 50 wt% of NMP; and (b) the second container comprises 35 wt% of degarelix acetate and 65 wt% NMP.

66. The pharmaceutical composition of any one of claims 62-65, wherein unreacted lactide and glycolide monomers in the PLG copolymer are less than about 1.0 wt%.

67. The pharmaceutical composition of any one of claims 62-65, wherein unreacted lactide and glycolide monomers in the PLG copolymer are less than about 0.5 wt%, than about 0.4 wt %, less than about 0.3 wt %, less than about 0.2 wt% and less than about 0.1 wt%.

68. A pharmaceutical composition, comprising: (a) about 10-30 wt% of degarelix acetate; (b) about 20-80 wt% of NMP; and (c) about 10-50 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of about 5-30 kDa.

69. The pharmaceutical composition of claim 68, wherein the composition comprises: (a) about 16 wt% of degarelix acetate; (b) about 57 wt% of NMP; and (c) about 27 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of about 19 kDa.

70. A pharmaceutical composition, comprising: (a) a first container comprising 30-70 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of 5-30 kDa, and 30-70 wt% of NMP; and (b) a second container comprising 20-40 wt% of degarelix acetate and 60-80 wt% NMP.

71. The pharmaceutical composition of claim 70, wherein: (a) the first container comprises 50 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of 19 kDa, and 50 wt% of NMP; and (b) the second container comprises 35 wt% of degarelix acetate and 65 wt% NMP.

72. A pharmaceutical composition, comprising: (a) about 10-30 wt% of degarelix acetate; (b) about 10-40 wt% of NMP; (c) about 10-40 wt% of DMSO; and (d) about 10-50 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of about 5-30 kDa.

73. The pharmaceutical composition of claim 72, wherein the composition comprises: (a) about 16 wt% of degarelix acetate; (b) about 30 wt% of NMP; (c) about 27 wt% of DMSO; and (d) about 27 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of about 19 kDa.

74. A pharmaceutical composition, comprising: (a) a first container comprising 30-70 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of 5-30 kDa, and 30-70 wt% of NMP; and (b) a second container comprising 20-40 wt% of degarelix acetate and 60-80 wt% DMSO.

75. The pharmaceutical composition of claim 74, wherein: (a) the first container comprises 50 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of 19 kDa, and 50 wt% of NMP; and (b) the second container comprises 35 wt% of degarelix acetate and 65 wt% DMSO.

76. A pharmaceutical composition, comprising: (a) about 10-30wt% of degarelix citrate; (b) about 20-80 wt% of NMP; and (c) about 10-50 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of about 5-30 kDa.

77. The pharmaceutical composition of claim 76, wherein the composition comprises: (a) about 16 wt% of degarelix citrate; (b) about 57 wt% of NMP; and (c) about 27 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of about 19 kDa.

78. A pharmaceutical composition, comprising: (a) a first container comprising 30-70 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of 19 kDa, and 50 wt% of NMP; and (b) a second container comprising 20-40 wt% of degarelix citrate and 60-80 wt% NMP.

79. The pharmaceutical composition of claim 78, wherein: (a) the first container comprises 50 wt% of 75:25 PLG copolymer having at least one hydroxyl end group and having a weight average molecular weight of 19 kDa, and 50 wt% of NMP; and (b) the second container comprises 35 wt% of degarelix citrate and 65 wt% NMP.

80. A method of treating prostate cancer in a subject, comprising subcutaneously administering the composition of any one of claims 1-79 to the subject.

81. The method of claim 80, wherein the prostate cancer is advanced prostate cancer.

82. The method of claim 80 or 81, wherein the dose amount of degarelix or pharmaceutically acceptable salt in the composition is administered in a dose amount of about 40 mg to about 500 mg.

83. A method of reducing serum testosterone levels in a subject to 50 ng/dL or less, comprising subcutaneously administering the composition of any one of claims 1-79 to the subject.

84. The method of claim 83, wherein serum testosterone levels are reduced to 20 ng/dL or less.

85. The method of claim 83, wherein serum testosterone levels are reduced to 10 ng/dL or less.

86. A method of suppressing ovarian function in a subject with hormone-receptor positive breast cancer, comprising subcutaneously administering the composition of any one of claims 1-79 to the subject.

87. The method of claim 86, wherein the hormone receptor positive breast cancer is estrogen receptor (ER) positive breast cancer.

88. The method of claim 87, wherein the subject’s estradiol (E2) production level is suppressed to a level less than about 20 pg/mL to about less than about 2 pg/mL.

89. The method of claim 86, wherein the subject’s follicle stimulating hormone (FSH) level is suppressed to a level less than about 40 IU/L.

90. The method of claim 86, wherein the subject’s luteinizing hormone (LH) level is suppressed to a level less than about 4 IU/L.

91. A method of treating central precocious puberty (CPP) in a subject comprising subcutaneously administering the composition of any one of claims 1-79 to the subject.

92. The method of claim 91, wherein the subject’s CPP blood serum LH concentration level is reduced to a pre-pubertal concentration level of less than about 4 IU/L.

93. The method of any one of claims 80-92, wherein the composition is administered about once every 1 month, about once every 2 months, about once every 3 months, about once every 4 months, about once every 5 months, or about once every six months.

94. The method of any one of claims 80-92, wherein the composition is administered to the subject about once every three months.

95. The method of any one of claims 80-92, wherein the composition is administered as a loading dose followed by a maintenance dose of the composition about 1 month, 2 months or 3 months after the loading dose has been administered.

96. The method of claim 95, when a loading dose is administered, the maintenance dose is administered every 1 month, every 2 months or every 3 months thereafter.

97. A prefilled syringe system for administration of the composition of claim 1, comprising a single syringe containing the composition of any one of claims 1-79.

98. The prefilled syringe system of claim 97, wherein the syringe is a mixing syringe.

99. The prefilled syringe system of claim 97, wherein the syringe is a dual-chambered syringe, and the composition of claim 1 is contained within one chamber of the syringe.

100. The prefilled syringe system of claim 97, wherein the syringe is a dual-chambered syringe, and wherein the biodegradable polymer and biocompatible solvent are contained in one chamber of the syringe and the degarelix or pharmaceutically acceptable salt is contained in the second chamber of the syringe.

101. The prefilled syringe system of claim 100, wherein the degarelix or pharmaceutically acceptable salt is lyophilized.

102. The prefilled syringe system of claim 100, wherein the degarelix or pharmaceutically acceptable salt is dissolved or suspended in the biocompatible solvent.

103. A prefilled syringe system for administration of the composition of any one of claims 1-79, comprising a first syringe containing the biodegradable polymer and the biocompatible solvent, and a second syringe containing the degarelix.

104. The prefilled syringe system of claim 103, wherein the first syringe and the second syringe are coupled together.

105. The prefilled syringe system of claim 103, wherein the degarelix or pharmaceutically acceptable salt is lyophilized.

106. The prefilled syringe system of claim 103, wherein the degarelix or pharmaceutically acceptable salt is dissolved or suspended in the biocompatible solvent.

107. A kit comprising the prefilled syringe system of any one of claims 97-106 and instructions.

108. A product comprising the composition of any one of claims 1-79 for use in a method of treating prostate cancer or CPP.

109. A product comprising the composition of any one of claims 1-79 for use in a method of reducing serum testosterone levels to 50 ng/dL or less.

110. A product comprising the composition of any one of claims 1-79 for use in a method of suppressing ovarian function in a subject with hormone-receptor positive breast cancer.