Sacituzumab govitecan formulations and methods of manufacture

Abstract

The present disclosure relates to methods and processes for manufacture of sacituzumab govitecan (SG), as well as formulations comprising SG.

Description

SACITUZUMAB GOVITECAN FORMULATIONS AND METHODS OFMANUFACTURECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of U.S. Provisional Application No. 63 / 654,737, filed May 31, 2024, the entire contents of which is incorporated herein by reference.FIELD

[0002] The present disclosure relates to formulations and methods of manufacture of sacituzumab govitecan (“SG”).BACKGROUND

[0003] Sacituzumab govitecan (“SG”) is a first-in-class antibody-drug-conjugate (“ADC”) comprised of a humanized monoclonal antibody (“hRS7”) that binds to the cell-surface receptor, Trop 2, a payload (“SN-38”) that is a topoisomerase I inhibitor, and a linker (“CL2A”), that couples the antibody to the payload. SG is marketed under the name TRODELVY®. The first U.S. approval for TRODELVY® was in 2020. TRODELVY® is approved in the U.S. for (1) unresectable locally advanced or metastatic triple-negative breast cancer in patients who have previously received two or more prior systemic therapies, at least one of them for metastatic disease; and (2) unresectable locally advanced or metastatic HR-positive, HER2- (IHC 0, IHC 1+, or IHC 2+ / ISH) breast cancer in patients who have received endocrine-based therapy and at least two additional systemic therapies in the metastatic setting. Due to the high unmet need in these patient populations, TRODELVY’ s use has been on the rise. To meet the demand, more TRODELVY® had to be produced. Thus, disclosed herein are processes and methods for manufacturing SG that permit a larger pharmaceutical batch size while maintaining stability. Additionally, disclosed herein are stable liquid and lyophilized formulations of SG.SUMMARY

[0004] Disclosed herein are stable liquid pharmaceutical formulations comprising about 15 mg / mL to about 70 mg / mL SG; a surfactant; and a buffer, wherein the pH of the formulation is about 5.5 to about 6.8. In certain embodiments, the formulation comprises a bulking agent. In certain embodiments, the bulking agent is selected from the group consisting of sucrose, trehalose, and mannitol. In certain embodiments, the buffer is selected from the group consisting of MES, histidine, phosphate, and maleate. In certain embodiments, the buffer is selected from the group consisting of MES and histidine. In certain embodiments the buffer is MES. In certain embodiments, the formulation is stable at about -20°C after storage for at least about 30 days. In certain embodiments, the formulation is stable at about <-20°C after storage for at least about 30days. In certain embodiments, DAR is about 7.4 or greater after storage at about -20°C for at least about 30 days. In certain embodiments, % HMW species is stable for at least about 30 days at about -20°C. In certain embodiments, the formulation is stable irrespective of SG concentration. In certain embodiments, DAR is about 7.3 or greater after storage at about 5°C for about 60 hours. In certain embodiments, DAR is stable for at least about 30 days as both a liquid formulation at about -20°C and as a lyophilized formulation at about 40°C (DAR is greater than 7.38).

[0005] Disclosed herein are stable lyophilized pharmaceutical formulations comprising 10 mg / kg to about 70 mg / kg SG; a surfactant; and a buffer, wherein pH of the formulation is 5.5 to a 6.8. In certain embodiments, the formulation comprises a bulking agent. In certain embodiments, the bulking agent is selected from the group consisting of sucrose, trehalose, and mannitol. In certain embodiments, the bulking agent is sucrose. In certain embodiments, the buffer is selected from the group consisting of MES, histidine, phosphate, and maleate. In certain embodiments, the buffer is selected from the group consisting of MES and histidine. In certain embodiments, the buffer is MES. In certain embodiments, the formulation is stable at 40°C after storage for at least 30 days. In certain embodiments, DAR is 7.4 or greater after storage for at least 30 days. In certain embodiments, the formulation comprises 40 mg / mL SG. In certain embodiments, the formulation is stable for at least 30 days as both a pre-lyophilized formulation at -20°C and as a lyophilized formulation at 40°C.

[0006] In certain embodiments, disclosed herein is a stable liquid formulation comprising about 15 mg / mL - about 40 mg / mL; a buffer; and a surfactant, wherein pH of the formulation is about 6.0 to about 7.0. In certain embodiments, the formulation comprises a bulking agent. In certain embodiments, the formulation is suitable for large scale production. In certain embodiments, the formulation comprises about 10 mM to about 100 mM buffer. In certain embodiments, the formulation comprises about 10 mM tolOO mM bulking agent. In certain embodiments, the formulation comprises 0.01 to 0.05 % w / v surfactant. In certain embodiments, the buffer is MES. In certain embodiments, the bulking agent is trehalose. In certain embodiments, the formulation comprises 20 mg / mL to 25 mg / mL SG. In certain embodiments, the formulation comprises 25 mg / mL SG. In certain embodiments, the formulation comprises 20 mM to 100 mM MES.

[0007] In certain embodiments, disclosed herein is a method of manufacture of SG, comprising the step of subjecting a mixture comprising a crude ADC through at least two different purification filters. In certain embodiments, one of the purification filters is UF / DF. In certain embodiments, one of the purification filters is activated carbon or a carbon filter. In certain embodiments, the method comprises a conjugation step prior to subjecting the mixture through at least two purification filters, wherein the conjugation step comprises a step of contacting an antibody composition comprising greater than 10 g / L hRS7 with drug linker, forming a conjugation mixturehaving greater than 6 mg / mL hRS7. In certain embodiments, the antibody composition comprises greater than about 30 g / L hRS7, and the conjugation mixture has greater than about 15 mg / mL hRS7.

[0008] In certain embodiments, disclosed herein is a method of manufacture of SG, comprising a conjugation step, wherein the conjugation step comprises a step of contacting an antibody composition comprising greater than 10 g / L hRS7 with drug linker, forming a conjugation mixture comprising greater than 6 mg / mL hRS7.

[0009] In certain embodiments, disclosed herein is a pre-lyophilization liquid formulation comprising: 10 mg / mL - 40 mg / mL of an ADC of the structure of Formula I and a pH of 5.8 to6.5, wherein the formulation is stable.

[0010] In certain embodiments, disclosed herein is a stable pre-lyophilization liquid formulation comprising: 10 mg / mL - 40 mg / mL of an ADC of the structure of Formula I and a pH of 5.8 to6.5, wherein the formulation comprises < 1.0 pg / mg free drug-related impurities as measured by the methods disclosed herein.

[0011] In certain embodiments, disclosed herein is a stable pre-lyophilization liquid formulation comprising: 10 mg / mL - 40 mg / mL of an ADC of the structure of Formula I and a pH of 5.8 to6.5, wherein the formulation comprises < 1.0 pg / mg free SN-38 as measured by the methods disclosed herein.

[0012] In certain embodiments, disclosed herein is a stable pre-lyophilization liquid formulation comprising: 10 mg / mL - 40 mg / mL of an ADC of the structure of Formula I and a pH of 5.8 to6.5, wherein the formulation comprises < 3.0 % HMW species as measured by the methods disclosed herein.

[0013] In certain embodiments, disclosed herein is a stable pre-lyophilization liquid formulation comprising: 10 mg / mL - 40 mg / mL of an ADC of the structure of Formula I; a buffer suitable to maintain pH of the formulation from 5.8 to 6.5; wherein the formulation has a DAR of about 7.6 as measured by native LC / MS.

[0014] In certain embodiments, disclosed herein is a stable pre-lyophilization liquid formulation comprising: 10 mg / mL to 40 mg / mL of an ADC of the structure of Formula I; a buffer; a bulking agent; and a surfactant; wherein pH of the formulation is 5.8 to 6.0.

[0015] In certain embodiments, disclosed herein is a stable pre-lyophilization liquid formulation comprising: 15 mg / mL - 40 mg / mL of an ADC of the structure of Formula I; a buffer; a bulking agent; and a surfactant; wherein pH of the formulation is 5.8 to 6.5.

[0016] In certain embodiments, disclosed herein is a pre-lyophilization liquid formulation of sufficient stability to limit free SN-38 to < 1.0 pg / mg for 1 month at 2-8°C as measured by themethods disclosed herein, wherein the formulation comprises a buffer that maintains pH of the pre-lyophilization liquid formulation from 5.8 to 6.0.

[0017] In certain embodiments, disclosed herein is a pre-lyophilization liquid formulation comprising: < 1.0 pg / mg free SN-38; < 1.0 pg / mg free drug-related impurities; < 3.0% HMW species; and a buffer that maintains pH of the formulation from 5.8 to 6.0; wherein the formulation is stable.

[0018] In certain embodiments, disclosed herein is a batch or lot of 1000 vials, each comprising a lyophilized formulation of 180 mg to 200 mg SG and a pharmaceutically acceptable carrier, wherein median or average free SN-38 of the batch or lot is < 1.0 pg / mg. In certain embodiments, the confidence interval (two-sided) is 90% or 95%. In certain embodiments, the vials are obtained from or obtainable from a 10 kg pharmaceutical batch.

[0019] In certain embodiments, disclosed herein is a stable lyophilized formulation comprising: 180 mg to 200 mg SG; and a pharmaceutically acceptable carrier; wherein the stable lyophilized formulation is obtained from or obtainable from a pharmaceutical batch of 10 kg SG. In certain embodiments, the pharmaceutical batch comprises < 1.0 free drug-related impurities at release.

[0020] In certain embodiments, disclosed herein is a reconstituted pharmaceutical formulation suitable for intravenous administration comprising: 10 mg / mL SG; wherein pH of the formulation is 5.8 to 6.0, or about 5.8 to about 6.0.

[0021] In certain embodiments, disclosed herein is s stable lyophilized pharmaceutical composition comprising: 180 mg to 200 mg of an ADC of the structure of Formula I and a pharmaceutically acceptable carrier, wherein the composition comprises <1.0; <0.9 <0.8; <0.7; <0.6; or < 0.5 pg / mg free SN-38 as measured by the methods disclosed herein at 6 months at 5°C.

[0022] In certain embodiments, disclosed herein is a lyophilized formulation of about 180 mg to 200 mg SG, the lyophilized formulation obtained by or obtainable from lyophilizing an aqueous formulation comprising SG and pharmaceutically acceptable excipients, wherein said formulation comprises a buffer that maintains pH during lyophilization from 5.8 to 6.3.

[0023] In certain embodiments, disclosed herein is a lyophilized formulation of about 180 mg to 200 mg SG, the lyophilized formulation obtained by or obtainable from lyophilizing an aqueous formulation of said SG and pharmaceutically acceptable excipients, wherein the aqueous formulation comprises 15 mg / mL to 40 mg / mL SG and a buffer that maintains pH during lyophilization from 5.8 to 6.5.

[0024] In certain embodiments, disclosed herein is a stable lyophilized formulation comprising 180 mg to 200 mg SG obtained from a 10 kg pharmaceutical batch of SG, wherein free SN-38 is < 1.0 pg / mg at batch release.

[0025] In certain embodiments, disclosed herein is a method of manufacturing an ADC represented by the structure shown in Formula I, wherein when the method is used to manufacture 10 kg SG, the free SN-38 at batch release is < 1.0 pg / mg.

[0026] In certain embodiments, disclosed herein is a lyophilized pharmaceutical formulation of about 180 mg to 200 mg SG, wherein the lyophilized pharmaceutical formulation is stable for 12 months at 2-8°C at a residual moisture content of < 3.7%.

[0027] In certain embodiments, disclosed herein is a pharmaceutical batch comprising: 10 kg SG; and < 1.0 pg / mg free drug-related impurities, and wherein the pharmaceutical batch yield is > 97%. In certain embodiments, the pharmaceutical batch is produced by running a composition comprising SG through at least two different filtration methods.

[0028] In certain embodiments, disclosed herein are stable drug substance formulations comprising 10 mg / mL SG - 70 mg / mL SG; wherein the formulations are stable < - 35°C for 18 - 36 months.

[0029] In certain embodiments, disclosed herein are stable formulations comprises 50 g / L - 60 g / L hRS7, wherein the formulations are stable at 2-8°C for 3 years and at < -30°C for > 24 months.

[0030] In certain embodiments, a pre-lyophilization liquid formulation drug substance comprising an ADC of the structure of Formula I and a means for stabilizing said ADC in said liquid formulation drug substance is provided (Linking Table 1).Linking Table 1.

[0031] In certain embodiments, a reconstituted liquid formulation drug product comprising an ADC of the structure of Formula I and a means for stabilizing said liquid formulation drug product is provided (Linking Table 2).Linking Table 2.

[0032] In certain embodiments, disclosed herein is a drug product comprising a lyophilized ADC comprising the ADC of Formula I; a reconstituted ADC in a liquid formulation drug product, and a means for achieving in said liquid formulation drug product a desired concentration of SN- 38 and a desired stability of said ADC (Linking Table 3).Linking Table 3.

[0033] In certain embodiments, disclosed herein is an ADC comprising a monoclonal humanized antibody means that bind to Trop-2; a cytotoxic topoisomerase inhibitor means, and a means for linking said cytotoxic means to said antibody means, wherein said ADC is capable of a concentration of 40 mg / ml in a liquid formulation, a pH of 5.8, and a lyophilization cycle of less than 4 days (Linking Table 4).Linking Table 4.

[0034] In certain embodiments, disclosed herein is a reconstituted liquid formulation drug product comprising a monoclonal humanized antibody means that bind to Trop-2; a cytotoxic topoisomerase inhibitor means, and means for linking said cytotoxic means to said antibody means; surfactant means; buffer means; and bulking agent means; wherein said liquid formulation drug product is capable of a concentration of 40 mg / ml in a liquid formulation; a pH of 5.8; a lyophilization cycle less than 4 days; stability at -20°C after storage for at least about 30 days; a DAR of 7.4 or greater after storage at -20°C for at least about 30 days; % HMW species stability for at least about 30 days at -20°C; a DAR of 7.3 or greater after storage at 5°C for about 60 hours;a stable DAR for at least about 30 days as both a liquid formulation at -20°C and as a lyophilized formulation at 40°C (Linking Table 5).Linking Table 5.

[0035] In certain embodiments, disclosed herein are methods of treating a human patient in need of treatment for (1) unresectable locally advanced or metastatic triple-negative breast cancer wherein said patient had previously received two or more systemic therapies, at least one of them for metastatic disease; and / or unresectable locally advanced or metastatic, HR+, HER2- (IHC 0, IHC 1+, or IHC 2+ / ISH-) breast cancer wherein said patient has received endocrine based therapy and at least two additional systemic therapies in the metastatic setting comprising: administering to said patient an amount effective for said treatment of a reconstituted liquid formulation drug product comprising an ADC of Formula I and a means for stabilizing said liquid formulation drug product (Linking Table 6).Linking Table 6.

[0036] In certain embodiments, disclosed herein are methods of treating a human patient in need of treatment for unresectable locally advanced or metastatic TNBC who are previously untreated in the metastatic setting comprising an ADC of Formula I and a means for stabilizing said liquid formulation drug product (Linking Table 7). In certain embodiments, the human patient is PD-Ll-negative. In certain embodiments, the human patient is PD-L1 positive and received previous PD-[L]-1 inhibitor therapy in the curative setting.Linking Table 7.

[0037] In certain embodiments, disclosed herein is a drug substance comprising an ADC of Formula I produced by filtering the crude ADC through a carbon filter.

[0038] In certain embodiments, disclosed herein is a drug product comprising an ADC of Formula I produced by filling a vial with less than 20 mL; < 19 mL; < 18 mL; < 17 mL; < 16 mL; < 15 mL; < 14 mL; < 13 mL; < 12 mL; < 11 mL; < 10 mL; < 9 mL; or < 8 mL of drug substance comprising SG.

[0039] In certain embodiments, disclosed herein is a reconstituted liquid drug product comprising an ADC of Formula I produced by filling a vial with less than 20 mL; < 19 mL; < 18 mL; < 17 mL; < 16 mL; < 15 mL; < 14 mL; < 13 mL; < 12 mL; < 11 mL; < 10 mL; < 9 mL; or < 8 mL of drug substance comprising SG, wherein the reconstituted liquid drug product comprises 10 mg / mL SG.

[0040] In certain embodiments, disclosed herein is A method of manufacturing a composition comprising SG, wherein the method comprises: (a) pooling mAb, wherein the mAb is hRS7, at a concentration greater than 10 g / L in a buffer; (b) reducing the pooled mAb for at least about 4 hours to produce reduced mAb; (c) conjugating the reduced mAb to produce a crude antibodydrug conjugate (ADC), wherein the conjugation mixture comprises greater than 6 mg / mL reduced mAb and about 10 molar equivalents of drug-linker, wherein the drug-linker is CL2A-SN-38; and (d) purifying SG from the crude ADC using at least two different purification methods. In certain embodiments, the step (a) comprises pooling mAb at a concentration of 60 g / L or 50 g / L.

[0041] In certain embodiments, wherein step (a) comprises pooling mAb at a concentration greater than 10 g / L in a buffer comprising sodium acetate. In certain embodiments, the crude ADC comprises 15 mg / mL SG - 45 mg / mL SG. In certain embodiments, the at least two different purification methods comprise passing the crude ADC through at least two different purification filters. In certain embodiments, SG is purified from the crude ADC by activated carbon filtration or by carbon depth filtration. In certain embodiments, SG is purified from the crude ADC by ultrafiltration / diafiltration. In certain embodiments, disclosed herein is a composition comprising SG obtainable by any of the preceding embodiments. In certain embodiments, the compositioncomprises greater than 10 mg / mL SG, such as 25 mg / mL SG. In certain embodiments, the composition comprises < 2.0 pg / mg free SN-38; < 1.9 pg / mg free SN-38; < 1.8 pg / mg free SN- 38; < 1.7 pg / mg free SN-38; < 1.6 pg / mg free SN-38; < 1.5 pg / mg free SN-38; <1.4 pg / mg free SN-38; < 1.3 pg / mg free SN-38; < 1.2 pg / mg free SN-38; < 1.1 pg / mg free SN-38; < 1.0 pg / mg free SN-38; < 0.9 pg / mg free SN-38; < 0.8 pg / mg; < 0.7 pg / mg free SN-38; < 0.6 pg / mg free SN- 38; < 0.5 pg / mg free SN-38; < 0.4 pg / mg free SN-38; < 0.3 pg / mg free SN-38; < 0.2 pg / mg free SN-38; or < 0.1 pg / mg free SN-38. In certain embodiments, free SN-38 is measured at drug substance release.

[0042] In certain embodiments, disclosed herein are pre-lyophilization liquid pharmaceutical formulations comprising an ADC of the structure as shown in Formula I, greater than 10 mg / mL SG, a surfactant, and a buffer, and wherein: the DAR of the formulation is no lower than 7.35 when the formulation is stored for at least 30 days at a temperature of -20°C; the formulation comprises less than 10 pg / mg free SN-38 when the formulation is stored for at least 30 days at a temperature of -20°C; and / or the free drug-related impurities (FDRI) are no more than 1.0 pg / mg. In certain embodiments, the formulation comprises 20 mg / mL SG to 40 mg / mL SG, such as 25 mg / mL SG. In certain embodiments, the surfactant is polysorbate 80 and the formulation comprises 0.02 % w / v to 0.04 %w / v polysorbate 80, such as about 0.025%w / v polysorbate 80. In certain embodiments, the buffer is MES and the formulation comprises 41 mM to 82 mM MES, such as 51 mM MES. In certain embodiments, the formulation also comprises trehalose, and the formulation comprises 50 nM to 100 nM trehalose, such as 62.5 nM trehalose. In certain embodiments, the buffer maintains a pH of 5.8 to 6.5. In certain embodiments, disclosed herein is a kit comprising less than 20 ml, such as less than 10 ml of the pre-lyophilization liquid pharmaceutical formulation of any one of the foregoing formulations in a 3 OR vial or a 20R vial. In certain embodiments, the 30R vial comprises at least 8 ml of the pre-lyophilization liquid pharmaceutical formulation. In certain embodiments, disclosed here is a method comprising lyophilizing the pre-lyophilization liquid pharmaceutical formulation described in the foregoing or herein in a 30R vial and / or a 20R vial to produce a 30R vial or a 20R vial comprising a lyophilized pharmaceutical formulation, wherein the lyophilized pharmaceutical formulation comprises about 180 mg to about 200 mg SG. In certain embodiments, disclosed herein is a stable lyophilized pharmaceutical formulation in a 3 OR vial or a 20R vial obtainable by the foregoing methods. In certain embodiments, the stable lyophilized pharmaceutical formulation in a 30R vial or a 20R vial comprises a pharmaceutically acceptable carrier.BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG. 1 shows drug-antibody ratio (DAR) using size-exclusion ultraperformance liquid chromatography with ultraviolet detection (UV-SE-UPLC) of the crude ADC (in-process sample after conjugation of hRS7 with CL2A-SN-38) as a function of reduction time and reduction temperature during the manufacture of SG. The circle represents reduction at 10°C, the diamond represents reduction at 15°C, and the triangle represents reduction at 20°C.

[0044] FIG. 2 shows DAR (UV-SE-UPLC) and % HMW species (UV-SE-UPLC) species of the crude ADC (in-process sample after conjugation) as a function of tris(2-carboxyethyl) phosphine (TCEP), pH, and drug linker stoichiometry.

[0045] FIG. 3 shows DAR (UV-SE-UPLC) as a function of temperature of the high hRS7 concentration conjugation mixture (in-process sample).

[0046] FIGS. 4A-4B show effect of hRS7 concentration in the conjugation mixture on DAR (UV-SE-UPLC) and % HMW species (UV-SE-UPLC) of the crude ADC (in-process sample). FIG. 4A shows effect of hRS7 concentration in the conjugation mixture on DAR of the crude ADC (in-process sample). FIG. 4B shows effect of hRS7 concentration in the conjugation mixture on % HMW species of the crude ADC (in-process sample).

[0047] FIG. 5 shows an exemplary process of production of sacituzumab govitecan (SG). This process is described in Example 1.

[0048] FIGS. 6A-6D show % HMW species by size exclusion chromatography (SEC) at 25°C (FIG 6A); 5°C (FIG. 6B); -20°C (FIG. 6C); -80°C (FIG. 6D) and time points for the IX, 2X, and 2.5X SG drug substance formulations (Example 4).

[0049] FIGS. 7A-7D show DAR by SEC at 25°C (FIG 7A); 5°C (FIG. 7B); -20°C (FIG. 7C); - 80°C (FIG. 7D) and time points for the IX, 2X, and 2.5X SG drug substance formulations (Example 4).

[0050] FIGS. 8A-8E show % HMW species by SEC-DAR, % main peak, and % LMW by SEC at different temperatures and timepoints for the IX, 2X, and 2.5X SG drug product formulations (Example 4).

[0051] FIGS. 9A-9C show DAR by SEC-DAR at different temperatures and timepoints for the IX, 2X, and 2.5X SG drug product formulations (Example 4).

[0052] FIGS. 10A-10B show % acidic and % basic species by cation exchange chromatography (CEX) at different temperatures and timepoints for the IX, 2X, and 2.5X drug product formulations (Example 4).

[0053] FIG. 11 shows the preparation of the different formulations in the Buffer Study (Example 6).

[0054] FIGS. 12A-12B show % HMW species and % free SN-38 (366 nm) of liquid formulations of SG (Example 6) at a temperature of 18°C and at different timepoints (T=0 to T=50 hours).

[0055] FIGS. 13A-13B show % HMW species and % free SN-38 of liquid formulations of SG (Example 6) at a temperature of 5°C and at different timepoints.

[0056] FIGS. 14A-14B show % HMW species and % free SN-38 of liquid formulations of SG (Example 6) at -20°C and -80°C at timepoints of T=0 and T=30 days.

[0057] FIGS. 15A-15B show % free SN-38 by SEC and free SN-38 by RP-HPLC (reverse phase HPLC) of liquid formulations of SG (Example 6) at 5°C at T=0 to T=15 days.

[0058] FIG. 16 shows the preparation of the IX, 2X, 3X, and 4X drug substance formulations as further described in Example 5.

[0059] FIGS. 17A-17C show % HMW species at different timepoints or freeze thaw cycles for the IX, 2X, 3X, and 4X drug substance formulations (Example 5).

[0060] FIGS. 18A-18C show DAR at different timepoints or freeze thaw cycles for the IX, 2X, 3X, and 4X drug substance formulations (Example 5).

[0061] FIGS. 19A-19C show % free SN-38 at various timepoints or freeze thaw (F / T) cycles for the IX, 2X, 3X, and 4X drug substance formulations (Example 5).

[0062] FIGS. 20A-20B show % HMW species and % free SN-38 at 40°C from T=0 to about T=45 for drug product formulations (Example 6).

[0063] FIGS. 21A-21B show % HMW species (Example 6) of the drug product formulations comprising (2-(N-morpholino)ethanesulfonic acid) (MES) or histidine (HIS) as buffers at various timepoints at 40°C.

[0064] FIGS. 22A-22B show free SN-38 (pg / mL) (FIG. 22 A) and % free SN-38 (FIG. 22B) for the drug product lyophilized formulations (Example 6) at 40°C at various timepoints.

[0065] FIGS. 23A-23D show stability of liquid drug substance formulations of SG (Example 7) as measured by % HMW species (FIG. 23 A), monomer % (FIG. 23B), DAR (FIG. 23C), and % released SN-38 (at 366 nm) (FIG. 23D) at 5°C using SE-UPLC.

[0066] FIGS. 24A-24D show stability of liquid drug substance formulations of SG (Example 7) as measured by % HMW species (FIG. 24A), monomer % (FIG. 24B), DAR (FIG. 24C), and % released SN-38 (at 366 nm) (FIG. 24D) at 12°C using SE-UPLC.

[0067] FIGS. 25A-25D show the stability of liquid drug substance formulations (Example 7) as measured by % HMW species (FIG. 25A), monomer % (FIG. 25B), DAR (FIG. 25C), and % released SN-38 (at 366 nm) (FIG. 25D) at 18°C.

[0068] FIGS. 26A-26D show the stability of liquid drug substance formulations (Example 7) as measured by % HMW species (FIG. 26A), monomer % (FIG. 26B), DAR (FIG. 26C), and % released SN-38 (at 366 nm) (FIG. 26D) at 25°C.

[0069] FIGS. 27A-27D show the stability of the liquid drug substance formulations as measured by % HMW species (FIG. 27A), monomer % (FIG. 27B), DAR (FIG. 27C), and % released SN- 38 (at 366 nm) (FIG. 27D) at -20°C following storage for 30 days.

[0070] FIGS. 28A-28D show % HMW species, monomer % DAR and % released free SN-38 for the lyophilized drug product formulations (identified by sucrose concentration to protein concentration ratio) at various primary drying temperatures (Example 7).

[0071] FIG. 29A shows % HMW species results for lyophilized drug product formulations stored at 40°C for up to 30 days. FIG. 29B shows monomer % results for the lyophilized drug product formulations stored at 40°C for up to 30 days. FIG. 29C shows DAR results for the lyophilized drug product formulations stored at 40°C for up to 30 days. FIG. 29D shows % released SN-38 for the lyophilized drug product formulations stored at 40°C for up to 30 days (Example 7).

[0072] FIG. 30 shows free SN-38 concentration (pg / mL) measured with RP-HPLC method for the lyophilized drug product formulations (Example 7) stored at 40°C for up to 30 days.

[0073] FIG. 31 shows free-drug related impurity (FDRI) clearance by ultrafiltration / diafiltration (UF / DF) alone when SG is made using the methods and processes disclosed herein.

[0074] FIG. 32 is an SEC-UPLC chromatographic analysis of a crude antibody drug conjugate, CEX elution fraction, and the carbon filtered pool. The crude ADC in this example was made using the methods and processes disclosed herein.

[0075] FIG. 33 shows structures of CL2A-SN-38 and major CL2A-SN-38 related species.

[0076] FIGS. 34A-34C show at what part in the process confirmation studies were conducted as discussed in Example 10. FIG. 34A shows the end-to-end process for drug substance to drug products for this Example 10. FIG. 34B shows in-use studies of (2) surfactant concentrations. FIG. 34C shows lyophilization cycle assessment of (2) sucrose concentrations.

[0077] FIGS. 35A-35C show % HMW and free SN-38 at various timepoints from during the process of drug substance to drug product as discussed in Example 10.

[0078] FIGS. 36A-36B show stability of the drug substance formulations of Example 10 at the temperature of -80°C.

[0079] FIGS. 37A-37B show stability of the drug product formulations of Example 10 at the temperature of 5°C.

[0080] FIGS. 38A-38B show stability of the drug substance formulations of Example 10 under stressed conditions of 5°C.

[0081] FIGS. 39A-39B show stability of the drug product formulations of Example 10 under stressed conditions of 40°C.

[0082] FIGS. 40A-40C show the change in water content from the initial water content range at 5°C (FIG. 40A); 25°C (FIG. 40B); and 40°C (FIG. 40C) during the residual moisture content study of Example 11. In order from top to bottom, the depicted lines represent Range 3, Range 2, Range 1, and Control / Control-Schott.

[0083] FIGS. 41A-41C show % HMW species at 5°C (FIG. 41A); 25°C (FIG. 41B); and 40°C (FIG. 41C) across moisture ranges during the residual moisture content study of Example 11.

[0084] FIGS. 42A-42B show free SN-38 at 25°C (FIG. 42A); and 40°C (FIG. 42B) across moisture ranges during the residual moisture content study of Example 11. In general, the top line represented Range 3, whereas the bottom line represented Range 1.

[0085] FIGS. 43A-43C show % HMW species of drug substance produced by Process 1 (represented by circles) and Process 2 (represented by squares and represented by “manufacturing scale” which is at 10 kg) at various timepoints at 25°C (FIG. 43A) and 40°C (FIG. 43B). Top four lines represent Process 1, whereas bottom five lines represent Process 2 or Manf scale (2).

[0086] FIG. 44 compares the free drug related impurities in the drug substance produced by the process in Example 1 and produced by the process in Example 16.

[0087] FIGS. 45A-45C show the effect of pH on protein concentration as measured by Solo VPE at 5°C (FIG. 45A); 25°C (FIG. 45B); and 40°C (FIG. 45C).

[0088] FIG. 46 shows the absorbance spectrum of SG drug substance.

[0089] FIGS. 47A-47C show the effect of pH on turbidity at 5°C (FIG. 47A); 25°C (FIG. 47B); and 40°C (FIG. 47C).

[0090] FIGS. 48A-48C show the effect of pH on % HMW species of SG at 5°C (FIG. 48A); 25°C (FIG. 48B); and 40°C (FIG. 48C).

[0091] FIGS. 49A-49C show the effect of pH on DAR at 5°C (FIG. 49A); 25°C (FIG. 49B); and 40°C (FIG. 49C).

[0092] FIG. 50 shows an alternate process for the manufacture of SG as described in Example 16.

[0093] FIGS. 51A-51B show subvisible particles in drug substance formulations as shown in Table 32 by high accuracy liquid particle counter (HIAC) on stability.

[0094] FIGS. 52A-52B show subvisible particles in drug product formulations as shown in Table 32 by HIAC on stability.

[0095] FIG. 53A (target concentration in IV bag 1.1 mg / mL) and FIG. 53B (target concentration in IV bag 3.4 mg / mL) show SVP levels (subvisible particles) of formula 2 (40 mg / mL protein in 20 mM His, 190 mM sucrose, 0.04% PS80 in pH 5.8 before and after infusion.

[0096] FIG. 54A (target concentration in IV bag 1.1 mg / mL) and FIG. 54B (target concentration in IV bag 3.4 mg / mL) show SVP levels (subvisible particles) of formula 3 (40 mg / mL protein in 20 mM His, 190 mM sucrose, 0.01% PS80 in pH 5.8 before and after infusion.

[0097] FIGS. 55A-55B show results of additional study 2 (formula 2 vs. T1 formulation - Table 33B). FIG. 55A shows SVP by HIAC and FIG. 55B shows % free SN-38 by SEC. The F2 formulation was comparable to the T1 formulation with respect to both.

[0098] FIGS. 56A-56B show % HMW species by SEC after dilution with saline (FIG. 56A) and in use (FIG. 56B).

[0099] FIG. 57 compares the degradation rate (as measured by free SN-38) of the drug substance produced by the process in Example 1 compared with the drug substance produced by the process in Example 16 under the stressed conditions of 2-8°C. Lines with filled symbols correspond to Process 1 (Example 16) and lines with hallowed symbols correspond to Process 2 (Example 1). Each line corresponds to a lot.

[0100] FIGS. 58A-58C show the SEC / DAR degradation rate for drug substance made by the process in Example 16 and the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. Lines with filled symbols correspond to Process 1 (Example 16) and lines with hallowed symbols correspond to Process 2 (Example 1). Each line corresponds to a lot.

[0101] FIGS. 59A-59B show rCE (reduced capillary electrophoresis-SDS) degradation rate assessment for drug substance made by the process in Example 16 and for the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. Lines with filled symbols correspond to Process 1 (Example 16) and lines with hallowed symbols correspond to Process 2 (Example 1). Each line corresponds to a lot.

[0102] FIGS. 60A-60C show CEX degradation rate assessment for drug substance made by the process in Example 16 and for the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. Lines with filled symbols correspond to Process 1 (Example 16) and lines with hallowed symbols correspond to Process 2 (Example 1). Each line corresponds to a lot.

[0103] FIGS. 61A-61B show potency degradation rate assessment for drug substance made by the process in Example 16 and for the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. Lines with filled symbols correspond to Process 1 (Example 16) and lines with hallowed symbols correspond to Process 2 (Example 1). Each line corresponds to a lot.DETAILED DESCRIPTIONAnti-TR0P2 ADC: Sacituzumab govitecan C‘SG ”)

[0104] SG is a pharmaceutical composition comprising an antibody-drug conjugate (“ADC”) comprised of (1) a drug (“SN-38”), a topoisomerase 1 inhibitor that is an active metabolite of irinotecan; (2) a linker (“CL2A”); and a humanized monoclonal antibody (“hRS7 IgGk” or “sacituzumab”). CL2A couples SN-38 to hRS7, which binds to Trop-2. As used herein, “hRS7” or “hRS7 IgGk” refers to “sacituzumab.”

[0105] In certain embodiments, hRS7 is described, e.g., in W02003074566, Figures 3 and 4, incorporated by reference in its entirety.

[0106] In certain embodiments, SG is an antibody-drug conjugate (ADC) as represented by Formula I as shown below.(Formula I)

[0107] In certain embodiments, SG is represented by Formula I, and hRS7 in Formula I comprises heavy chain CDRs SEQ ID NO.: 13, SEQ ID NO.: 14, and SEQ ID NO.: 15; and light chain CDRs SEQ ID NO.: 16, SEQ ID NO.: 17, and SEQ ID NO.: 18 (as shown in Table 1).

[0108] In certain embodiments, SG is represented by Formula I, and hRS7 in Formula I comprises heavy chain as shown in SEQ ID NO.: 1 and light chain as shown in SEQ ID NO.: 2 (as shown in Table 1). In certain embodiments, the hRS7 antibody in SG comprises two heavy chains each having the sequence as shown in SEQ ID NO.: 1, and two light chains each having the sequence as shown in SEQ ID NO.: 2.

[0109] In certain embodiments, SG is represented by Formula I, and hRS7 in Formula I comprises the heavy chain variable region as shown in SEQ ID NO.: 19 and the light chain variable region as shown in SEQ ID NO.: 20.

[0110] In certain embodiments, the ADC comprises drug molecules linked to the antibody moieties in various stoichiometric molar ratios depending on the configuration of the antibody and, at least in part, the method used to effect configuration. In certain embodiments, the drugantibody ratio (“DAR”) is about 7.6. In certain embodiments, DAR is about 7.8. In certain embodiments, the DAR is about 8.0 The DAR is determined using the methods as disclosed herein.Table 1. Amino Acid Sequences of hRS7 antibodyDefinitions[OHl] The singular forms “a,” “an,” and “the” include the plural referents unless the context dictates otherwise.

[0112] As used herein, ranges and amounts can be expressed as “about” a particular value or range. About also includes the exact amount. Hence “about 5 pL” means “about 5 pL” and also “5 pL.” Unless otherwise stated, “about” includes an amount that would be expected to be to be within experimental error, i.e., within 5% of the stated value.

[0113] “Antibody-drug conjugate” or “ADC” refers to SG, i.e., (hRS7 or sacituzumab-CL2A- SN-38). As used herein, “crude ADC” refers to SG prior to purification. In certain embodiments, ADC as used herein refers to SG, i.e., the compound of Formula I.

[0114] “Batch” or “pharmaceutical batch” refers to pre-lyophilized drug substance. A batch size, for example, of 6.6 kg refers to 6.6 kg of SG. In certain embodiments, “batch” refers to a specific quantity of drug that is intended to have uniform character and quality. In certain embodiments, the “batch” is produced according to a single manufacturing order during the same cycle of manufacture. For example, a 10 kg batch of SG refers to the production of 10 kg SG via a single manufacturing run. The actual yield of SG may be different, i.e., may be 97% of 10 kg, for example.

[0115] “Bulking agent” as used herein, can refer to a cryoprotectant and / or a filler.

[0116] “Free drug-related impurities” (“FDRI”) as used herein, includes species such as, but not limited to, TCEP-CL2A-SN38; reduced CL2A-SN38; cyclic CL2A-SN38; CL2A-SN38 (drug linker); and CL2A-SN38 dimer. In certain embodiments, FDRI does not comprise free SN-38. In certain embodiments, FDRI is a process-related impurity, introduced / formed during conjugation. In certain embodiments, FDRI refers to SN-38-related impurities. In certain embodiments, the FDRI comprise about 20-40% free SN-38 by weight; about 25-35% by weight; or about 27% by weight free SN-38. In certain embodiments, conjugatable impurities contain a maleimide that may react with an antibody. FDRI comprises SN-38 which is not conjugated to the mAb but may either conjugatable or non-conjugatable. Not all conjugatable impurities are FDRI (and vice versa). In certain embodiments, FDRI is measured at using reverse phase HPLC.

[0117] “Free SN-38” or “free SN38” is a degradation product of SG’s drug linker and is a process and product related impurity. In certain embodiments, “free SN-38” is described in pg / mg units. This refers to pg free SN-38 per mg total protein. In certain embodiments, free SN-38 is a product related impurity. In certain embodiments, free SN-38 (pg / mg) refers to amount (pg) of free SN-38 per mg protein concentration, i.e., per mg mAb.

[0118] “Drug substance” as used herein, refers to a liquid formulation prior to its lyophilization. It may also be referred to as “pre-lyophilization liquid formulation” or “pre-lyophilization liquid pharmaceutical formulation” or versions thereof. It is to be understood that the drug substance may be frozen for storage, but may still be considered a “liquid formulation” as described herein.

[0119] “Drug product” as used herein, refers to a lyophilized formulation or a reconstituted formulation.

[0120] “High concentration conjugation mixture” or “high concentration mAh concentration mixture,” as used herein, refers to a conjugation mixture comprising mAh and drug-linker that has > about 6 mg / mL mAh; > 6 mg / mL mAh; > about 7 mg / mL mAb; > 7 mg / mL mAb; > about 8 mg / mL mAb; > 8 mg / mL mAb; > about 9 mg / mL mAb; > 9 mg / mL mAb; > than about 10 mg / mL mAb; or > 10 mg / mL mAb. In certain embodiments, “mAb” refers to (“hRS7 IgGk” or “sacituzumab) .”

[0121] “Large scale production” or “bulk production” as used herein refers to a production scale that is greater than or equal to about 6.5 kg. In certain embodiments, this refers to a production scale greater than 6.5 kg.

[0122] “Liquid formulation” or “pre-lyophilization liquid formulation” as used herein, refers to a pharmaceutical formulation intended to be lyophilized. It is to be understood that the liquid formulation may be stored in frozen conditions, and thus may be “frozen” but still referred to as “liquid formulation” or some variation thereof.

[0123] “Pharmaceutically acceptable” (such as the recitation of “pharmaceutically excipient”) refers to a material that is compatible with administration to a human subject, i.e., the material does not cause an undesirable biological effect. Examples of pharmaceutically acceptable excipients are described in the “Handbook of Pharmaceutical Excipients,” Rowe et al., Ed. (Pharmaceutical Press, 7thEd., 2012).

[0124] “Plurality” refers to “more than one.”

[0125] “Released SN-38” as used herein, is the same as “free SN-38.”

[0126] “Release” i.e., “at release” as used herein, refers to time 0 (T=0). For example, for a measurement to be taken at drug substance “release” refers to the measurement being taken immediately after the drug substance is produced.

[0127] “ Stable,” as used herein, refers to, in certain embodiments, in the context of a drug product as disclosed herein, refers to a drug product having sufficient stability for storage and use as a lyophilized, and subsequently reconstituted prior to use, pharmaceutical product. In certain embodiments, in the context of a drug substance as disclosed herein, “stable” refers to a drug substance having sufficient stability for storage and use to produce a lyophilized, and subsequently reconstituted prior to use, pharmaceutical product. In certain embodiments, “stable” as used herein, refers to a drug substance or a drug product as disclosed herein having sufficient stability to be suitable for pharmaceutical use as an intravenous infusion. In certain embodiments, “stable” refers to meeting the current acceptance standards for TRODELVY®.Impurities and Important Attributes of SG

[0128] In certain embodiments, compositions comprising hRS7 and / or SG have impurities.

[0129] For example, low molecular weight (LMW) species comprise clipped as well as incomplete molecules. Incomplete molecules are missing full chains [(e.g. heavy light (HL), heavy heavy (HH)] whereas clips contain truncated chains. Generally, they pose similar risks to biological activity and PK. Clipped species may be detected by rCE-SDS. High levels of the HL and HH impurities may cause loss of active molecules, resulting in reduced TROP2 binding.

[0130] As another example, compositions comprising hRS7 and / or SG may have aggregation. Protein aggregation refers to high molecular weight (HMW) species forming clusters of low solubility material that can turn into sub-visible or visible particles. Aggregates may have an impact on biological activity as well as immunogenicity. For example, forced degradation photo stress caused a maximum increase of HMW species (up to 25%), and resulted in reduced binding to TROP2 and loss of cytotoxicity.

[0131] In certain embodiments, drug linker hydrolysis results in loss of SN-38 and may impact both biological activity by changing DAR and safety by increase of free SN-38. Hydrolysis of drug linker may be monitored by the CEX / IdeS assay, where the Fab with hydrolyzed drug linker acquires negative charge and manifests in Fab acidic variants. In certain embodiments, for example, the relative abundance of Fab acidics under thermal stress at 40°C for 5 weeks increased to 87%; at low pH for 7 days increased to 84%; high pH for 7 days increased to 89%; while the drug product control had a Fab acidics peak area of 34%. Intact mass analysis confirmed Fab acidics are enriched in hydrolyzed drug linker.

[0132] In certain embodiments, the composition comprising SG comprises free SN-38. Free SN- 38 is measured because it can be easily hydrolyzed from the linker and affect biological activity by lowering the DAR and safety. It may be measured by SEC-HPLC. The degradation and release of SN-38 is part of the mechanism-of-action for SG, and the and half-life of SN-38 in serum / solution is 1 day. Free SN-38 may be inversely related to DAR. An increase of free SN-38 may be observed with a decrease in DAR. Loss of SN-38 impacts the biological activity.

[0133] SN-38 is a topoisomerase I inhibitor, hence untargeted free SN-38 release may cause systemic toxic effects. SG’s CL2A linker is moderately stable, resulting in moderate kinetics of SN-38 release (72 hours) and half-life of 1 day in serum. These features allow SG to be targeted to the tumor site and reduces untargeted effects. Release of SN-38 is the mechanism-of-action of SG, but premature release in the blood stream can potentially cause toxicity.

[0134] Another important quality attribute for SG is DAR. DAR is important to control because the amount of conjugated drug on the antibody is directly linked to the biological activity. It may be measured by SEC-UPLC and HIC-HPLC. DAR may be impacted by all the forced degradationconditions. In certain embodiments, for example, the DAR decreased to 6.8 at 22°C for 3 days (thermal stress). Under photo stress (8300 lux 1.1 W / m2 UV / Vis light, 5 - 10°C), a non-linear decay of DAR was observed, and it reduced to ~5 in 3 days. At high pH, 50% loss of DAR was observed in a 7-day period. Binding to TROP2 and cytotoxicity were impacted under light stress. DAR is very important for potency.

[0135] Free drug-related impurities (FDRI) is an impurity.

[0136] RP-HPLC may be used to quantify the amount of CL2A-SN-38 and related species. FDRI may include i) TCEP-CL2A-SN-38; ii) reduced CL2A-SN-38, iii) cyclic CL2A-SN-38 and iv) free CL2A-SN-38.Pharmaceutical batches, compositions, and formulations of SG

[0137] Disclosed herein are pharmaceutical batches, compositions, or formulations of SG. In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations of SG that comprise less than about 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, or 1 pg / mg free SN-38.

[0138] In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations of SGthat comprise less than about 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, or 1 pg / mg free drug-related impurities.

[0139] In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations of SG that comprise less than about 0.10 pg / mg, 0.9 pg / mg, 0.8 pg / mg, 0.7 pg / mg, 0.6 pg / mg, 0.5 pg / mg, 0.4 pg / mg, 0.3 pg / mg, 0.2 pg / mg, or 0.1 pg / mg free SN-38.

[0140] In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations of SG that comprise less than 0.10 pg / mg, 0.9 pg / mg, 0.8 pg / mg, 0.7 pg / mg, 0.6 pg / mg, 0.5 pg / mg, 0.4 pg / mg, 0.3 pg / mg, 0.2 pg / mg, or 0.1 pg / mg free SN-38.

[0141] In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations of SG that comprise less than about 0.10 pg / mg, 0.9 pg / mg, 0.8 pg / mg, 0.7 pg / mg, 0.6 pg / mg, 0.5 pg / mg, 0.4 pg / mg, 0.3 pg / mg, 0.2 pg / mg, or 0.1 pg / mg free drug related impurities.

[0142] In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations of SG that comprise less than 0.10 pg / mg, 0.9 pg / mg, 0.8 pg / mg, 0.7 pg / mg, 0.6 pg / mg, 0.5 pg / mg, 0.4 pg / mg, 0.3 pg / mg, 0.2 pg / mg, or 0.1 pg / mg free drug related impurities. Pharmaceutical Batches

[0143] In certain embodiments, disclosed herein is a pharmaceutical batch comprising SG and impurities, such as free-drug related impurities or free SN-38.

[0144] In certain embodiments, the pharmaceutical batches as disclosed herein are at the 6.0 kg;6.5 kg; 7 kg; 7.5 kg; 8 kg; 8.5 kg; 9 kg; 9.5 kg; 10 kg; 10.5 kg; 11 kg; 11.5 kg; 12 kg; 12.5 kg; 13kg; 13.5 kg or 14 kg scale. In certain embodiments, the yield of SG of the foregoing batches is 95%; 96%; 97%; 98%; 99%; 100%; 101%; 102%; 103%; or 104%.

[0145] In certain embodiments, free SN-38 and free drug-related impurities are measured at drug substance release (i.e., TO or time 0 or end of production time). In certain embodiments, these are measured at various timepoints as shown in Example 17 Table 46.

[0146] In certain embodiments, the pharmaceutical batch comprises SG and less than about 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg of free-drug related impurities. In certain embodiments, batch comprises free drug-related impurities is the LOQ (limit of quantification).

[0147] In certain embodiments, the pharmaceutical bath comprises 10 kg SG and less than 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg of free-drug related impurities. In certain embodiments, batch comprises free drug-related impurities is the LOQ (limit of quantification). In certain embodiments, this is measured at drug substance release. In certain embodiments, the pharmaceutical bath comprises greater than 6.5 kg SG and less than 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg of free-drug related impurities. In certain embodiments, batch comprises free drug-related impurities is the LOQ (limit of quantification). In certain embodiments, this is measured at drug substance release.

[0148] In certain embodiments, the pharmaceutical bath comprises greater than 6.5 kg SG and less than 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg of free SN-38. In certain embodiments, this is measured at drug substance release.

[0149] In certain embodiments, the pharmaceutical bath comprises 10 kg SG and less than 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg free SN-38. In certain embodiments, this is measured at drug substance release.

[0150] In certain embodiments, a pharmaceutical product comprising SG is produced from the pharmaceutical batch. In certain embodiments, the pharmaceutical product is produced after storage of the pharmaceutical batch. As such, the pharmaceutical batch is of sufficient stability to be stored prior to production of the pharmaceutical product. In certain embodiments, thepharmaceutical product is a drug substance. In certain embodiments, the pharmaceutical product is the drug product.

[0151] In certain embodiments, the pharmaceutical batch comprises or consists essentially of at least about 6.5 kg SG and less than about 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg free, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg free SN-38 and / or free drug-related impurities. In certain embodiments, free SN-38 is measured by RP-HPLC. In certain embodiments, the yield of SG is at least 97%.

[0152] In certain embodiments, disclosed herein is a pharmaceutical batch comprising at least about 6.6 kg SG and less than about 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg free SN-38 and / or free drug-related impurities.

[0153] In certain embodiments, disclosed herein is a pharmaceutical batch comprising at least about 6.7 kg SG and less than about 10 pg / mg, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or 0.1 pg / mg of free SN-38 and / or free drug-related impurities.

[0154] In certain embodiments, the batch comprises free drug-related impurities at the LOQ (limit of quantification).

[0155] In certain embodiments, disclosed herein is a pharmaceutical batch having less than about 2 pg / mg free drug related impurities, wherein the pharmaceutical batch comprises at least about 1 kg of SG. In certain embodiments, disclosed herein is a pharmaceutical batch having less than about 10 pg / mg free drug related impurities, wherein the pharmaceutical batch comprises at least about 2 kg SG; at least about 3 kg SG; at least about 4 kg SG; at least about 5 kg SG; or at least about 6 kg SG. In certain embodiments, disclosed herein is a pharmaceutical batch having less than about 10 pg / mg free drug related impurities, wherein the pharmaceutical batch comprises at least about 6.1 kg SG; at least about 6.2 kg SG; at least about 6.3 kg SG; at least about 6.4 kg SG; at least about 6.5 kg SG; at least about 6.6. kg SG; at least about 6.7 kg SG; at least about 6.8 kg SG; at least about 6.9 kg SG; or at least about 7.0 kg SG.

[0156] In certain embodiments, disclosed herein is a pharmaceutical batch having less than about 10 pg / mg free drug related impurities, wherein the pharmaceutical batch comprises at least about 1 kg of SG. In certain embodiments, disclosed herein is a pharmaceutical batch having less than about 10 pg / mg free drug -related impurities, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or less than about 0.1 pg / mg free drug-related impurities; wherein the pharmaceutical batch comprises at least about 2 kg SG; at least about 3 kg SG; at leastabout 4 kg SG; at least about 5 kg SG; or at least about 6 kg SG. In certain embodiments, disclosed herein is a pharmaceutical batch having less than about 10 pg / mg free drug-related impurities, 9 pg / mg, 8 pg / mg, 7 pg / mg, 6 pg / mg, 5 pg / mg, 4 pg / mg, 3 pg / mg, 2 pg / mg, 1 pg / mg, 0.9 pg / mg, 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; or less than about 0.1 pg / mg free drug related impurities, wherein the pharmaceutical batch comprises at least about 6.1 kg SG; at least about 6.2 kg SG; at least about 6.3 kg SG; at least about 6.4 kg SG; at least about 6.5 kg SG; at least about 6.6. kg SG; at least about 6.7 kg SG; at least about 6.8 kg SG; at least about 6.9 kg SG; or at least about 7.0 kg SG.

[0157] In certain embodiments, the pharmaceutical batch has less than about 10 pg / mg free drug related impurities; less than about 9 pg / mg free drug related impurities; less than about 8 pg / mg free drug related impurities; less than about 7 pg / mg free drug related impurities; less than about 6 pg / mg free drug related impurities; less than about 5 pg / mg free drug related impurities; less than about 4 pg / mg free drug related impurities; less than about 3 pg / mg free drug related impurities; less than about 2 pg / mg free drug related impurities; or less than about 1 pg / mg free drug related impurities. In certain embodiments, the free drug-related impurities is at the LOQ (limit of quantification).Pharmaceutical Compositions or Formulations

[0158] In certain embodiments, disclosed herein are pharmaceutical compositions or formulations comprising SG and < about 10 pg / mg free SN-38; < about 9 pg / mg free SN-38; < about 8 pg / mg free SN-38; < about 7 pg / mg free SN-38; < about 6 pg / mg free SN-38; < about 5 pg / mg free SN-38; < about 4 pg / mg free SN-38; < about 3 pg / mg free SN-38; < about 2 pg / mg free SN-38; or < about 1 pg / mg free SN-38.

[0159] In certain embodiments, the pharmaceutical formulations comprise SG and < 1.0 pg / mg;< 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 04 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg or < 0.1 pg / mg free SN-38. In certain embodiments, this is measured at drug substance release, drug product release, or on stability (drug substance or drug product).

[0160] In certain embodiments, the pharmaceutical formulations comprise SG and < 1.0 pg / mg;< 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 04 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg or < 0.1 pg / mg free drug-related impurities. In certain embodiments, this is measured at drug substance release.

[0161] In certain embodiments, free SN-38 is measured by RP-HPLC. In certain embodiments, the pharmaceutical compositions or formulations are liquid compositions or formulations. In certain embodiments, the pharmaceutical compositions or formulations are lyophilized compositions or formulations. In certain embodiments, the pharmaceutical compositions or formulations are stable.

[0162] In certain embodiments, disclosed herein are pharmaceutical compositions or formulations comprising SG and less than about 10 pg / mg free drug related impurities. In certain embodiments, the pharmaceutical compositions or formulations are liquid compositions or formulations. In certain embodiments, the pharmaceutical compositions or formulations are lyophilized compositions or formulations. In certain embodiments, the pharmaceutical compositions or formulations are stable.Vials

[0163] In certain embodiments, disclosed herein is a vial comprising a stable lyophilized pharmaceutical formulation comprising about 180 mg to about 200 mg of SG (or about 180 mg or about 200 mg) and less than about 10 pg / mg free drug related impurities.

[0164] Each vial comprises about 180 mg to about 200 mg of SG, and each vial is obtained from, or obtainable from, a pharmaceutical batch.

[0165] In certain embodiments, disclosed herein is a vial comprising a stable lyophilized pharmaceutical formulation comprising about 180 mg to about 200 mg SG and < about 9 pg / mg free SN-38; < about 8 pg / ml free SN-38; < about 7 pg / mg free SN-38; < about 6 pg / mg free SN-38; < about 5 pg / mg free SN-38; < about 4 pg / mg free SN-38; < about 3 pg / mg free SN-38; < about 2 pg / mg free SN-38; < about 1 pg / mg free SN-38; < about 0.9 pg / mg free SN-38; < about 0.8 pg / mg free SN-38; < about 0.7 pg / mg free SN-38; < about 0.6 pg / mg free SN-38; < about 0.5 pg / mg free SN-38; < about 0.4 pg / mg free SN-38; < about 0.3 pg / mg free SN-38; < about 0.2 pg / mg free SN-38; or < about 0.1 pg / mg free SN-38. In certain embodiments, free SN-38 is measured at drug product release. In certain embodiments, free SN-38 is measured on stability for up to 36 months.

[0166] In certain embodiments, disclosed herein is a vial comprising a stable lyophilized pharmaceutical formulation comprising 180 mg to 200 mg SG and < 9 pg / mg free SN-38; < 8 pg / ml free SN-38; < 7 pg / mg free SN-38; < 6 pg / mg free SN-38; < 5 pg / mg free SN-38; < 4 pg / mg free SN-38; < 3 pg / mg free SN-38; < 2 pg / mg free SN-38; < 1 pg / mg free SN-38; < 0.9 pg / mg free SN-38; < 0.8 pg / mg free SN-38; < 0.7 pg / mg free SN-38; < 0.6 pg / mg free SN-38; < 0.5 pg / mg free SN-38; < 0.4 pg / mg free SN-38; < 0.3 pg / mg free SN-38; < 0.2 pg / mg free SN- 38; or < 0.1 pg / mg free SN-38. In certain embodiments, free SN-38 is measured at drug product release. In certain embodiments, free SN-38 is measured on stability for up to 36 months.

[0167] In certain embodiments, the vial is a 30R vial. In certain embodiments, the vial is a 20R vial. In certain embodiments, the vial is a 50R vial. In certain embodiments, the vial comprises less than about 20 mL of drug substance pre-lyophilization. In certain embodiments, the vial comprises less than about 10 mL of drug substance pre-lyophilization. In certain embodiments, the drug product disclosed herein is produced from less than 20 mL of drug substance as disclosedherein. In certain embodiments, the vial comprises about 8 mL of drug substance prelyophilization. In certain embodiments, the vial comprises sufficient volume of drug substance to produce about 180 mg to about 200 mg SG post-lyophilization. In certain embodiments, the vial comprises 8 mL of drug substance pre-lyophilization. In certain embodiments, the vial comprises sufficient volume of drug substance to produce 180 mg to 200 mg SG post-lyophilization.

[0168] In certain embodiments, disclosed herein is a stable lyophilized formulation, comprising about 180 mg to about 200 mg SG; and a pharmaceutically acceptable carrier; wherein the stable lyophilized formulation is obtained from a plurality of pharmaceutical batches wherein the plurality of batches have a median free SN-38, wherein the median free SN-38 is less than about 10 pg / mg; less than about 9 pg / mg; less than about 8 pg / mg; less than about 7 pg / mg; less than about 6 pg / mg; less than about 5 pg / mg; less than about 4 pg / mg; less than about 3 pg / mg; less than about 2 pg / mg; less than about 1 pg / mg; less than about 0.9 pg / mg; less than about 0.8 pg / mg; less than about 0.7 pg / mg; less than about 0.6 pg / mg; less than about 0.5 pg / mg; less than about 0.4 pg / mg; less than about 0.3 pg / mg; less than about 0.2 pg / mg; or less than about 0.1 pg / mg free SN-38.

[0169] In certain embodiments, disclosed herein is a stable lyophilized formulation, comprising about 180 mg to about 200 mg SG; and a pharmaceutically acceptable carrier; wherein the stable lyophilized formulation is obtained from a plurality of vials wherein the plurality of vials have a median free SN-38, wherein the median free SN-38 is < about 10 pg / mg; < about 9 pg / mg; < about 8 pg / mg; < about 7 pg / mg; < about 6 pg / mg; < about 5 pg / mg; < about 4 pg / mg; < about 3 pg / mg; < about 2 pg / mg; < about 1 pg / mg; < about 0.9 pg / mg; < about 0.8 pg / mg; < about 0.7 pg / mg; < about 0.6 pg / mg; < about 0.5 pg / mg; < about 0.4 pg / mg; < about 0.3 pg / mg; < about 0.2 pg / mg; or < about 0.1 pg / mg free SN-38.

[0170] In certain embodiments, disclosed herein is a plurality of vials, wherein each of the vials comprises a lyophilized formulation as disclosed herein, and wherein the plurality of vials are obtainable from, or obtained from, a 10 kg pharmaceutical batch of SG.

[0171] In certain embodiments, disclosed herein is a stable lyophilized formulation, comprising about 180 mg to about 200 mg SG; and a pharmaceutically acceptable carrier; wherein the stable lyophilized formulation is obtained from or obtainable from a pharmaceutical batch of about at least about 40 g; at least about 400 g (pilot scale); at least about 1 kg SG; at least about 2 kg SG; at least about 3 kg SG; at least about 4 kg SG; at least about 5 kg SG; or at least about 6 kg SG; wherein the batch has a median free SN-38, wherein the median free SN-38 is less than about 10 pg / mg; less than about 9 pg / mg; less than about 8 pg / mg; less than about 7 pg / mg; less than about 6 pg / mg; less than about 5 pg / mg; less than about 4 pg / mg; less than about 3 pg / mg; less than about 2 pg / mg; less than about 1 pg / mg; less than about 0.9 pg / mg; less than about 0.8pg / mg; less than about 0.7 pg / mg; less than about 0.6 pg / mg; less than about 0.5 pg / mg; less than about 0.4 pg / mg; less than about 0.3 pg / mg; less than about 0.2 pg / mg; or less than about 0.1 pg / mg free SN-38. In certain embodiments, the stable lyophilized formulation is obtained from or obtainable from a pharmaceutical batch of 10 kg SG; wherein the batch has a free SN-38 of < 0.6 pg / mg as measured at drug substance release.

[0172] In certain embodiments, disclosed herein is a stable lyophilized formulation, comprising 180 mg to 200 mg SG; and a pharmaceutically acceptable carrier; wherein the stable lyophilized formulation is obtained from or obtainable from a pharmaceutical batch of at least 40 g; at least 400 g (pilot scale); at least 1 kg SG; at least 2 kg SG; at least 3 kg SG; at least 4 kg SG; at least 5 kg SG; or at least 6 kg SG; wherein the batch has a median free SN-38, wherein the median free SN-38 is < 10 pg / mg; < 9 pg / mg; < 8 pg / mg; < 7 pg / mg; < 6 pg / mg; < 5 pg / mg; < 4 pg / mg; < 3 pg / mg; < 2 pg / mg; <1 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; or < 0.1 pg / mg free SN-38. In certain embodiments, the stable lyophilized formulation is obtained from or obtainable from a pharmaceutical batch of 10 kg SG. In certain embodiments, free SN-38 is measured at drug substance release and / or drug product release. In certain embodiments, free SN-38 is measured on stability for up to 36 months. For example, the lyophilized formulations may be reconstituted at various timepoints to measure free SN-38 at various timepoints.

[0173] In certain embodiments, disclosed herein is a plurality of pharmaceutical batches or vials, wherein the plurality of pharmaceutical batches or vials each comprise a lyophilized formulation comprising SG, wherein the plurality of batches or vials have a median free SN-38 of about less than or equal to about 10 pg / mg. In certain embodiments, the confidence interval is 95% that the test criteria (free SN-38) can remain within the acceptance criteria for shelf life.

[0174] In certain embodiments, disclosed herein is a batch or lot of 200 or more vials each comprising a lyophilized formulation comprising SG, wherein the median or average free SN-38 of the lyophilized formulations in the batch or lot is < 1.0 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; <0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; < 0.1 pg / mg free SN-38 at drug product release and / or on stability for up to 36 months. In certain embodiments, the vials are obtained from or obtainable from a pharmaceutical batch of 10 kg SG.

[0175] In certain embodiments, disclosed herein is a batch or lot of 500 or more vials each comprising a lyophilized formulation comprising SG, wherein the median or average free SN-38 of the lyophilized formulations in the batch or lot is < 1.0 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; <0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; < 0.1 pg / mg free SN-38 at drug product release and / or on stability for up to 36 months. In certain embodiments, the vials are obtained from or obtainable from a pharmaceutical batch of 10 kg SG.

[0176] In certain embodiments, disclosed herein is a batch or lot of 1000 or more vials each comprising a lyophilized formulation comprising SG, wherein the median or average free SN-38 of the lyophilized formulations in the batch or lot is < 1.0 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; <0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; < 0.1 pg / mg free SN-38 at drug product release and / or on stability for up to 36 months. In certain embodiments, the vials are obtained from or obtainable from a pharmaceutical batch of 10 kg SG.

[0177] In certain embodiments, disclosed herein is a batch or lot of 2000 each comprising a lyophilized formulation comprising SG, wherein the median or average free SN-38 of the lyophilized formulations in the batch or lot is < 1.0 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; <0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; < 0.1 pg / mg free SN- 38 at drug product release and / or on stability for up to 36 months. In certain embodiments, the vials are obtained from or obtainable from a pharmaceutical batch of 10 kg SG.

[0178] In certain embodiments, disclosed herein is a stable lyophilized formulation comprising about 180 mg to about 200 mg SG and free SN-38, and a pharmaceutically acceptable carrier, wherein the stable lyophilized formulation is obtained from a plurality of batches, pilot batches, or vials, wherein median free SN-38 of the batches, pilot batches, or vials is less than or equal to about 10 pg / mg. In certain embodiments, disclosed herein is a stable lyophilized formulation comprising about 180 mg to about 200 mg SG and free drug-related impurities, and a pharmaceutically acceptable carrier, wherein the stable lyophilized formulation is obtained from a plurality of batches, pilot batches, or vials, wherein free drug-related impurities is < to about 10 pg / mg; < about 9 pg / mg; < about 8 pg / mg; < about 7 pg / mg; < about 6 pg / mg; < about 5 pg / mg;< about 4 pg / mg; < about 3 pg / mg; < about 2 pg / mg; < about 1 pg / mg; < about 0.9 pg / mg; < about 0.8 pg / mg; < about 0.7 pg / mg; < about 0.6 pg / mg; or < 0.5 pg / mg. In certain embodiments, the vials are obtained from or obtainable from a pharmaceutical batch of 10 kg SG.

[0179] In certain embodiments, disclosed herein is a stable lyophilized formulation comprising 180 mg to 200 mg SG and free SN-38, and a pharmaceutically acceptable carrier, wherein the stable lyophilized formulation is obtained from a plurality of batches, pilot batches, or vials, wherein median free SN-38 of the batches, pilot batches, or vials is less than or equal to 10 pg / mg. In certain embodiments, disclosed herein is a stable lyophilized formulation comprising 180 mg to 200 mg SG and free drug-related impurities, and a pharmaceutically acceptable carrier, wherein the stable lyophilized formulation is obtained from a plurality of batches, pilot batches, or vials, wherein free drug-related impurities is < 10 pg / mg; < 9 pg / mg; < 8 pg / mg; < 7 pg / mg; < 6 pg / mg; < 5 pg / mg; < 4 pg / mg; < 3 pg / mg; < 2 pg / mg; < 1 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg;< 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; < 0.1 pg / mg. In certain embodiments, the vials are obtained from or obtainable from a pharmaceutical batch of10 kg SG. In certain embodiments, the free drug-related impurities are measured at drug substance release.DAR

[0180] In certain embodiments, disclosed herein are pharmaceutical batches, compositions, and formulations comprising SG and having a DAR, an average DAR, or a median DAR of about 6.7 to about 8.0 as measured by the methods disclosed herein. In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations comprising SG and having a DAR, an average DAR, or a median DAR of about 6.7 to about 7.8 as measured by the methods disclosed herein. In certain embodiments, disclosed herein is a pharmaceutical formulation comprising about 180 mg of SG and having a DAR, an average DAR, or a median DAR of about 6.7 to about 7.8 as measured by the methods disclosed herein. In certain embodiments, disclosed herein is a pharmaceutical formulation comprising about 200 mg of SG and having a DAR, an average DAR, or a median DAR of about 6.7 to about 7.8 as measured by the methods disclosed herein. In certain embodiments, the pharmaceutical batches, compositions, or formulations comprising SG have a DAR of about 6.7 to about 7.8 as measured by size-exclusion ultraperformance liquid chromatography (SE-UPLC) with ultraviolet (UV) detection at 280 nm and 366 nm as disclosed herein. In certain embodiments, disclosed herein are pharmaceutical batches, compositions, and formulations comprising SG and having a DAR, an average DAR, or a median DAR of 6.7 to 8.0 as measured by the methods disclosed herein. In certain embodiments, disclosed herein are pharmaceutical batches, compositions, or formulations comprising SG and having a DAR, an average DAR, or a median DAR of 6.7 to 7.8 as measured by the methods disclosed herein.

[0181] In certain embodiments, disclosed herein is a stable lyophilized formulation comprising about 180 mg to about 200 mg SG, and a pharmaceutically acceptable excipient, wherein the stable lyophilized formulation has a DAR of about 6.7 to about 7.8 as measured by SE-UPLC with UV detection at 280 nm and 366 nm as disclosed herein.

[0182] In certain embodiments, disclosed herein is a stable lyophilized formulation comprising 180 mg to 200 mg SG, and a pharmaceutically acceptable excipient, wherein the stable lyophilized formulation has a DAR of 6.7 to 7.8 as measured by SE-UPLC with UV detection at 280 nm and 366 nm as disclosed herein. In certain embodiments, the stable lyophilized formulation comprising 180 mg to 200 mg SG has a DAR of 7.4 to 7.8 at release, and on stability at intended storage conditions for 36 months.

[0183] In certain embodiments, disclosed herein is a pharmaceutical composition comprising SG; wherein at least about 50% of the SG molecules in the composition have a DAR of 8.Methods and Processes to Produce SG at Large-Scale

[0184] In certain embodiments, the methods and processes disclosed herein enable a greater amount of SG to be made per pharmaceutical batch while at least maintaining stability.

[0185] In certain embodiments, the methods disclosed herein enable manufacture of SG at a pharmaceutical batch size, for example, of greater than about 6.5 kg of SG while at least maintaining stability. In certain embodiments, the methods disclosed herein enable manufacture of SG at a pharmaceutical batch size of greater than, for example, about 6.6 kg of SG. In certain embodiments, the methods disclosed herein enable manufacture of SG at about 6 kg, about 7 kg, about 8 kg, about 9 kg, about 10 kg, about 11 kg, about 12 kg, about 13 kg, about 14 kg, or about 14 kg or greater of SG. In certain embodiments, the methods disclosed herein enable manufacture of SG at a pharmaceutical batch size of about 6.6 kg or greater. In certain embodiments, the methods disclosed herein enable manufacture of SG at a pharmaceutical batch size of about 10 kg. In certain embodiments, the methods disclosed herein enable manufacture of a pharmaceutical batch size of about 14 kg.

[0186] Certain aspects of the methods disclosed herein enable manufacture of more SG per pharmaceutical batch, such as increasing the concentration of the mAb, drug-linker, etc. used to make SG. For example, in certain embodiments, the concentration of the drug substance (prelyophilization) is increased (greater than 10 mg / mL). Increasing the concentration (i.e., greater than 10 mg / mL) of the drug substance enables manufacture of more SG per pharmaceutical batch. For example, when the drug substance has an SG concentration of 25 mg / mL (as opposed to 10 mg / mL), the amount of drug substance produced is 2.5X when placed in the same tank size.

[0187] As another example, in certain embodiments, the concentration of the mAb (i.e., hRS7) is increased (i.e., greater than about 6 mg / mL or greater than 6 mg / mL) in the conjugation mixture, which enables manufacture of more SG per pharmaceutical batch.

[0188] In certain embodiments, it is to be understood that one or more of the method or process steps disclosed herein can be omitted but still produce a drug substance or a drug product that is stable and has acceptable impurity levels. In certain embodiments, the methods and processes disclosed herein comprise multiple steps that enable scale up and minimize impurities. In certain embodiments, one or more of these steps may be omitted and the method may still enable scale up and minimize impurities. In certain embodiments, one or more of the method and process steps as disclosed herein may be eliminated and / or replaced, and still enable manufacture of a large pharmaceutical batch size, i.e., greater than about 6.5 kg of SG.

[0189] In certain embodiments, disclosed herein are methods of manufacturing a composition comprising SG. In certain embodiments, the methods comprise the step of pooling mAb (i.e., hRS7) at a concentration greater than about 10 g / L in a buffer such as PBS or sodium acetate. In certain embodiments, the pooled mAb has 50 to 60 g / L mAb in sodium acetate. In certainembodiments, the methods comprise the step of reducing the pooled Ab for at least about 4 hours. In certain embodiments, the methods comprise the step of conjugation, wherein the conjugation mixture comprises greater than about 6 mg / mL mAb (i.e., hRS7) and about 10 molar equivalents of drug-linker. In certain embodiments, the conjugation mixture comprises about 30 mg / mL mAb and about 10 molar equivalents of drug-linker. In certain embodiments, the methods comprise the step of producing a crude ADC. In certain embodiments, the crude ADC comprises free drug- related impurities and SG. In certain embodiments, the crude ADC comprises greater than about 30 pg / mg free drug-related impurities. In certain embodiments, the methods comprise the step of purifying the crude ADC using at least two different purification methods. In certain embodiments, the two different purification methods comprise carbon filtration and UF / DF. In certain embodiments, the methods comprise the step of purifying the crude ADC using carbon filtration or activated carbon. In certain embodiments, after the step of purifying the crude ADC using carbon filtration, but prior to the step of purifying using UF / DF, the crude ADC comprises less than about 0.10 pg / mg of free drug-related impurities. In certain embodiments, the methods comprise the step of purifying the crude ADC using UF / DF. In certain embodiments, the step of purifying the crude ADC using UF / DF comprises the step of using less than about 30 diafiltration volumes (DV). In certain embodiments, the methods comprise the step of purifying the crude ADC using CEX (cation exchange chromatography). In certain embodiments, the step of purifying the crude ADC using UF / DF comprises the step of using about 10 diafiltration volumes (DV). In certain embodiments, the step of purifying the crude ADC using UF / DF comprises the step of using about 9 diafiltration volumes (DV). In certain embodiments, using both carbon filtration and UF / DF to purify the crude ADC reduces the diafiltration volumes required to remove the free drug-related impurities. In certain embodiments, the activated carbon is CR40. For example, as shown in FIG. 31, about 46 diafiltration volumes may be required to clear the free drug-related impurities in the crude ADC using only UF / DF for purification. As another example, and as shown in FIG. 32, SEC-UPLC chromatographic analysis of the crude ADC, CEX elution fraction, and the carbon-filtered pool shows incomplete removal of free drug-related impurities by CEX and reduction of free drug related impurities by carbon filtration. In certain embodiments, the methods comprise the step of producing a drug substance formulation comprising about 25 mg / mL SG. In certain embodiments, the methods comprise the step of producing a drug substance formulation of about 20 mg / mL SG.Process Details mAb Preparation and Reduction

[0190] Pooled mAb solution or mAb input: In certain embodiments, after preparation, hRS7 is pooled from one or more lots (“pooled mAb solution” or “mAb input”). In certain embodiments,one or more filters are used to pool the mAb solution. Exemplary filters include, but are not limited to, a sterilizing grade PES (polyethersulfone) filter.

[0191] In certain embodiments, the pooled mAb solution comprises about 10 g / L hRS7 to about 100 g / L hRS7. In certain embodiments, the pooled mAb solution comprises greater than about 10 g / L hRS7. In certain embodiments, the pooled mAb solution comprises about 20 g / L hRS7 to about 90 g / L hRS7. In certain embodiments, the pooled mAb solution comprises about 25 g / L hRS7 to about 80 g / L hRS7. In certain embodiments, the pooled mAb solution comprises about 30 g / L hRS7 to about 70 g / L hRS7. In certain embodiments, the pooled mAb solution comprises about 40 g / L hRS7 to about 60 g / L hRS7. In certain embodiments, the pooled mAb solution comprises about 40 g / L hRS7, 50 g / L hRS7, 60 g / L hRS7, or 70 g / L hRS7. In certain embodiments, the pooled mAb solution comprises about 60 g / L hRS7. In certain embodiments, the pooled mAb solution / formulation comprises 60 g / L hRS7. In certain embodiments, the pooled mAb solution / formulation comprises 50 g / L hRS7.

[0192] In certain embodiments, the pooled mAb solution has a pH of about 5.0 to about 7.0. In certain embodiments, the pooled mAb solution has a pH of about 5.5 to about 6.5. In certain embodiments, the pooled mAb solution has a pH of about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0. In certain embodiments, the pooled mAb solution has a pH of about 5.7. In certain embodiments, the pooled mAb solution has a pH of 5.7.

[0193] In certain embodiments, the pooled mAb solution comprises about 60 g / L hRS7 and a buffer. In certain embodiments, the pooled mAb solution comprises about 60 g / L hRS7 and sodium acetate. In certain embodiments, the pooled mAb solution comprises about 60 g / L of hRS7 (mAb) and about 0.1 M sodium acetate at a pH of about 5.7.

[0194] In certain embodiments, the pooled mAb solution comprises EDTA. In certain embodiments, the pooled mAb solution comprises MES. In certain embodiments, the pooled mAb solution comprises sodium acetate. In certain embodiments, the pooled mAb solution comprises about 10 g / L hRS7 (mAb) and about 0.04 M PBS at a pH of about 7.4.

[0195] In certain embodiments, the pooled mAB solution comprising about 60 g / L of hRS7 is stable and is stored at 2-8°C.

[0196] Conditioned mAb solution: In certain embodiments, after pooling, but prior to reduction, the pooled mAb solution is conditioned with dilution buffer to achieve target pH and buffer matrix (“conditioned mAb solution”). In certain embodiments, the pooled mAb solution is conditioned in 0.2M MESZEDTA. In certain embodiments, the conditioned mAb solution target pH is about 6.3 to about 6.8. In certain embodiments, the conditioned mAb solution target pH is about 6.6 + 0.2. In certain embodiments, the conditioned mAb solution target pH is less than about 6.6. In certain embodiments, the conditioned mAb solution target pH is greater than about 6.6. In certainembodiments, the mAb solution is conditioned in another buffer matrix to achieve the target pH. In certain embodiments, the conditioned mAb solution does not comprise phosphate.

[0197] In certain embodiments, the excipients and pH of the conditioned mAb solution are dictated by the concentration of hRS7. For example, in certain embodiments, if the concentration of hRS7 is less than about 15 g / L, i.e., 10 g / L, then the solution may comprise a higher pH and / or phosphate. For example, if the hRS7 concentration is at about 10 g / L, then the conditioned mAb solution may comprise 0.1 M NaPO4 and 0.2 M EDTA at a pH of about 6.5.

[0198] Reduced mAb solution: In certain embodiments, the mAb solution or conditioned mAb solution is reduced, for example, to reduce interchain disulfide bonds that can be used as potential conjugation sites for the payload (“reduced mAb solution”). For example, if all four interchain disulfide bonds in the mAb are reduced, then (8) thiol groups are available for conjugating drug molecules. In certain embodiments, efficacy of the reduction process is determined by the subsequent DAR of the ADC. Time, temperature, pH, reaction time, and / or stoichiometry of the reduction process may be varied to determine the most efficacious reduction parameters to achieve the desired DAR.

[0199] In certain embodiments, the reduction reaction time is about 240 (4 hours) minutes to about 600 minutes. In certain embodiments, the reduction reaction time is 180 - 1140 minutes. In certain embodiments, the reduction reaction time is at least about 4 hours. As shown in FIG. 1, DAR is affected by reduction time.

[0200] In certain embodiments, the reduction time is about 180 minutes to about 360 minutes. In certain embodiments, the reduction time is about 180 minutes.

[0201] In certain embodiments, the reduction temperature is about 15°C to about 25°C. In certain embodiments, the reduction temperature is at least about 15°C. In certain embodiments, the reduction temperature is about 20°C + / - 2°C. As shown in FIG. 1, DAR is affected by temperature during the reduction process.

[0202] In certain embodiments, the conditioned mAb solution is reduced with about 5.5 equivalents of tris(2-carboxyethyl)phosphine (TCEP-HCL). In certain embodiments, the conditioned mAb solution is reduced with about 5.5 equivalents of TCEP in WFI. In certain embodiments, the conditioned mAb is reduced with 4-8 equivalents of TCEP in WFI. In certain embodiments, the TCEP concentration is reduced to lower the competitive reaction of TCEP and CL2A-SN-38 during the conjugation process.

[0203] In certain embodiments, the reduced mAb solution comprises about 10 g / L hRS7 to about 100 g / L hRS7. In certain embodiments, the reduced mAb solution comprises greater than about 10 g / L hRS7. In certain embodiments, the reduced mAb solution comprises about 20 g / L hRS7 to about 90 g / L hRS7. In certain embodiments, the reduced mAb solution comprises about 25 g / LhRS7 to about 80 g / L hRS7. In certain embodiments, the reduced mAb solution comprises about 30 g / L hRS7 to about 70 g / L hRS7. In certain embodiments, the reduced mAb solution comprises about 40 g / L hRS7 to about 60 g / L hRS7. In certain embodiments, the reduced mAb solution comprises about 40 g / L hRS7, 50 g / L hRS7, 60 g / L hRS7, or 70 g / L hRS7. In certain embodiments, the reduced mAb solution comprises about 60 g / L hRS7. In certain embodiments, the reduced mAb solution comprises 60 g / L hRS7. In certain embodiments, the reduced mAb solution comprises 50 g / L hRS7.Conjugation of mAb to drug-linker to produce ADC

[0204] In certain embodiments, the conjugation mixture (conjugating mAb with the drug-linker) comprises a concentration greater than about 6 mg / mL of mAb (hRS7). In certain embodiments, the conjugation mixture is a high concentration mAb conjugation mixture. In certain embodiments, the conjugation mixture (conjugating mAb with the drug-linker) comprises a concentration greater than 6 mg / mL of mAb (hRS7).

[0205] When a higher concentration of mAb is added to the conjugation mixture, this enables conjugation at a much higher concentration of mAb (and thus a higher concentration of druglinker is added), which enables production of a larger pharmaceutical batch of SG. For example, in certain embodiments, if the reduced mAb solution comprises about 60 g / L (60 mg / mL) mAb, then the mAb concentration in the conjugation mixture is about 30 g / L. In certain embodiments, the reduced mAb solution comprises a concentration of mAb that is greater than about 10 mg / mL mAb. In certain embodiments, the mAb concentration in the conjugation mixture is 30 g / L.

[0206] In certain embodiments, the mAb concentration in the conjugation mixture is greater than about 8 g / L. In certain embodiments, the mAb concentration in the conjugation mixture is between about 20 g / L and about 50 g / L. In certain embodiments, the mAb concentration in the conjugation mixture is about 10 g / L; about 15 g / L; about 20 g / L; about 25 g / L; about 35 g / L; about 40 g / L, about 45 g / L; or about 50 g / L. In certain embodiments, the mAb concentration in the conjugation mixture is about 30 g / L.

[0207] In certain embodiments, the mAb concentration in the conjugation mixture is greater than 8 g / L. In certain embodiments, the mAb concentration in the conjugation mixture is between 20 g / L and 50 g / L. In certain embodiments, the mAb concentration in the conjugation mixture is 10 g / L; 15 g / L; 20 g / L; 25 g / L; 35 g / L; 40 g / L, 45 g / L; or 50 g / L. In certain embodiments, the mAb concentration in the conjugation mixture is 30 g / L.

[0208] In certain embodiments, the pH of the conjugation mixture is about 6.0 to about 7.0 In certain embodiments, the pH of the conjugation mixture is about 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0. In certain embodiments, the pH of the conjugation mixture is about 6.3.

[0209] In certain embodiments, disclosed herein is a conjugation mixture suitable for producing a larger pharmaceutical batch of SG, comprising about 20 g / L to about 50 g / L hRS7 in the conjugation mixture. This concentration conjugation mixture is suitable for large scale SG production, as it maintains the appropriate DAR (FIG. 4 A) while limiting the generation of HMW species (FIG. 4B). In certain embodiments, the conjugation comprises about 30 g / L hRS7. In certain embodiments, the pH of the conjugation mixture is less than about 6.5. In certain embodiments, the pH of the conjugation mixture is about 6.3.

[0210] In certain embodiments, and as shown in FIG. 2, pH, concentration of TCEP, and drug linker stoichiometry may affect DAR and / or % HMW species of the crude ADC. In certain embodiments, increasing pH of the conjugation mixture increases % HMW species, but achieves a higher (and more favorable) DAR.

[0211] In certain embodiments, and as shown in FIG. 3, the conjugation mixture suitable for large scale pharmaceutical batch production as disclosed herein produces a drug substance with an average DAR of about 7.0-8.0 at a conjugation temperature of about 20°C-30°C.

[0212] In certain embodiments, the conjugation mixture does not comprise NEM (N-ethyl maleimide).

[0213] In certain embodiments, conjugation produces a crude ADC at a concentration of about 15-45 mg / mL SG. In certain embodiments, conjugation produces a crude ADC at a concentration of about 30 mg / mL SG.Carbon Depth Filtration

[0214] In certain embodiments, to enable production of larger pharmaceutical batches of SG, but to limit impurities such as free-drug related impurities associated with scale-up, activated carbon or carbon depth filtration is used to purify the ADC, i.e., to purify the crude ADC mixture after conjugation.

[0215] In certain embodiments, the crude ADC is filtered using activated carbon or carbon depth filtration. In certain embodiments, the crude ADC is filtered using activated carbon or carbon depth filtration after conjugation but prior to ultrafiltration / diafiltration. In certain embodiments, the crude ADC is purified using ultrafiltration / diafiltration prior to filtration using activated carbon or carbon depth filtration. In certain embodiments, carbon depth filtration is not used to purify the crude ADC. In certain embodiments, carbon depth filtration removes impurities, such as free-drug related impurities, created during the scale-up process. For example, the carbon filter removes and / or reduces unconjugated CL2A-SN-38 drug-linker. In certain embodiments, the carbon filter is a single stage depth filter. Exemplary carbon depth filtration process parameters are described in Example 1.UF / DF

[0216] In certain embodiments, after carbon depth filtration, the ADC (i.e., SG) is buffer exchanged via ultrafiltration / diafiltration prior to formulation. In certain embodiments, after UFDF, the filtered ADC is diluted and adjusted with a concentrated conditioning solution to achieve the target drug substance concentration. In certain embodiments, the target drug substance concentration is about 10 mg / mL; about 15 mg / mL; about 20 mg / mL; about 25 mg / mL; about 30 mg / mL; about 35 mg / mL; or about 40 mg / mL. In certain embodiments, the target drug substance concentration is 25 mg / mL. In certain embodiments, the target drug substance concentration is 10 mg / mL; 15 mg / mL; 20 mg / mL; 25 mg / mL; 30 mg / mL; 35 mg / mL; or 40 mg / mL. In certain embodiments, the target drug substance concentration is 25 mg / mL.Liquid Pharmaceutical Formulations Comprising SG (Drug Substance)

[0217] In certain embodiments, disclosed herein are liquid pharmaceutical formulations comprising SG. In certain embodiments, the liquid pharmaceutical formulations are prelyophilization (i.e., the liquid formulations are intended to be lyophilized) pharmaceutical formulations (i.e., “pre-lyophilization liquid formulations”). In certain embodiments, the liquid pharmaceutical formulation comprises about 10 mg / mL SG to about 70 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 10 mg / mL SG to 70 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises about 10 mg / mL SG to about 40 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 10 mg / mL SG to 40 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises greater than about 10 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises greater than 10 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises about 15 mg / mL to about 70 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 15 mg / mL to 70 mg / mL SG.In certain embodiments, the liquid pharmaceutical formulation comprises about 15 mg / mL SG to about 40 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 15 mg / mL SG to 40 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises about 10 mg / mL to about 25 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 10 mg / mL to 25 mg / mL SG.In certain embodiments, the liquid pharmaceutical formulation comprises about 15 mg / mL SG, about 20 mg / mL SG, about 25 mg / mL SG, about 30 mg / mL SG, about 35 mg / mL SG, and / or about 40 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises about 20 mg / mL to about 25 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises about 25 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 15 mg / mL SG, 20 mg / mL SG, 25 mg / mL SG, 30 mg / mL SG, 35 mg / mLSG, and / or 40 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 20 mg / mL to 25 mg / mL SG. In certain embodiments, the liquid pharmaceutical formulation comprises 25 mg / mL SG.

[0218] In certain embodiments, the pharmaceutical formulations disclosed herein are stable.

[0219] In certain embodiments, disclosed herein are stable, pre-lyophilization liquid pharmaceutical formulations comprising SG. In certain embodiments, the stable pre- lyophilization liquid pharmaceutical formulation comprises about 10 mg / mL SG to about 70 mg / mL SG. In certain embodiments, the stable pre-lyophilization liquid pharmaceutical formulation comprises about 10 mg / mL SG to about 40 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises greater than about 10 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises about 15 mg / mL SG to about 40 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises about 15 mg / mL SG, about 20 mg / mL SG, about 25 mg / mL SG, about 30 mg / mL SG, about 35 mg / mL SG, and / or about 40 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises about 25 mg / mL SG. In certain embodiments, the stable pre-lyophilization liquid pharmaceutical formulation comprises 10 mg / mL SG to 70 mg / mL SG. In certain embodiments, the stable pre-lyophilization liquid pharmaceutical formulation comprises 10 mg / mL SG to 40 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises greater than 10 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises 15 mg / mL SG to 40 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises 15 mg / mL SG, 20 mg / mL SG, 25 mg / mL SG, 30 mg / mL SG, 35 mg / mL SG, and / or 40 mg / mL SG. In certain embodiments, the stable, pre-lyophilization liquid pharmaceutical formulation comprises 25 mg / mL SG.

[0220] In certain embodiments, the stable, pre-lyophilization pharmaceutical formulation comprises about 10 mg / mL to about 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent (or filler). In certain embodiments, the stable pharmaceutical formulation comprises about 10 mg / mL to about 40 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the stable pharmaceutical formulation comprises about 15 mg / mL to about 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises about 15 mg / mL to about 40 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the stable pharmaceutical formulation does not comprise a surfactant. In certain embodiments, the stable pharmaceutical formulation does not comprise a bulking agent.

[0221] In certain embodiments, the stable, pre-lyophilization pharmaceutical formulation comprises 10 mg / mL to 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent (or filler). In certain embodiments, the stable pharmaceutical formulation comprises 10 mg / mL to 40 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the stable pharmaceutical formulation comprises 15 mg / mL to 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises 15 mg / mL to 40 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the stable pharmaceutical formulation does not comprise a surfactant. In certain embodiments, the stable pharmaceutical formulation does not comprise a bulking agent.

[0222] In certain embodiments, the stable pharmaceutical formulation comprises about 20 mg / mL to about 25 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises about 20 mg / mL to about 25 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the stable pharmaceutical formulation comprises about 25 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises about 25 mg / mL SG, a surfactant, and a buffer.

[0223] In certain embodiments, the stable pharmaceutical formulation comprises 20 mg / mL to 25 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises 20 mg / mL to 25 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the stable pharmaceutical formulation comprises 25 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises 25 mg / mL SG, a surfactant, and a buffer.

[0224] In certain embodiments, the pre-lyophilization pharmaceutical formulation comprises about 10 mg / mL to about 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the pharmaceutical formulation comprises about 10 mg / mL to about 40 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the pharmaceutical formulation comprises about 15 mg / mL to about 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the pharmaceutical formulation comprises about 15 mg / mL to about 40 mg / mL SG, a surfactant, and a buffer.

[0225] In certain embodiments, the pre-lyophilization pharmaceutical formulation comprises 10 mg / mL to 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the pharmaceutical formulation comprises 10 mg / mL to 40 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the pharmaceutical formulation comprises 15 mg / mL to 40 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the pharmaceutical formulation comprises 15 mg / mL to 40 mg / mL SG, a surfactant, and a buffer.

[0226] In certain embodiments, the pharmaceutical formulation comprises about 20 mg / mL to about 25 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the stable pharmaceutical formulation comprises about 20 mg / mL to about 25 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the pharmaceutical formulation comprises about 25 mg / mL SG, a surfactant, a buffer, and / or a bulking agent. In certain embodiments, the pharmaceutical formulation comprises about 25 mg / mL SG, a surfactant, and a buffer.

[0227] In certain embodiments, the pharmaceutical formulations as disclosed herein do not comprise a bulking agent. In certain embodiments, the pharmaceutical formulations as disclosed herein do not comprise a buffer. In certain embodiments, the pharmaceutical formulations as disclosed herein do not comprise a surfactant.

[0228] In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 15 mg / mL SG to about 70 mg / mL SG, a surfactant, a buffer, and a bulking agent. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 15 mg / mL to about 70 mg / mL SG, a surfactant, a buffer, and a bulking agent.

[0229] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 0.01 %w / v to about 0.1 %w / v surfactant. In certain embodiments, the pharmaceutical formulations disclosed herein comprise about 0.01 %w / v to about 0.025 %w / v surfactant. In certain embodiments, the formulations disclosed herein comprise about 0.01; 0.02; 0.025; 0.03; or 0.04 %w / v surfactant. In certain embodiments, the surfactant is polysorbate 80 (PS80). In certain embodiments, the surfactant is polysorbate 20 (PS20). In certain embodiments, the formulations disclosed herein comprise 0.01; 0.02; 0.025; 0.03; or 0.04 % w / v surfactant.

[0230] In certain embodiments, the buffer is selected from the group consisting of MES, succinate, citrate, histidine, phosphate, and maleate. In certain embodiments, the buffer is selected from MES and histidine. In certain embodiments, the buffer is MES. In certain embodiments, the buffer is histidine. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 10 mM to about 100 mM buffer. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise less than about 10 mM buffer or greater than about 100 mM buffer. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 10 mM buffer, about 20 mM buffer, about 30 mM buffer, about 40 mM buffer, about 50 mM buffer, about 60 mM buffer, about 70 mM buffer, about 80 mM buffer, about 90 mM buffer, or about 100 mM buffer. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 50 mM buffer. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 51 mM MES. In certain embodiments, the pharmaceutical formulations comprise more than one buffer. In certain embodiments, the pharmaceutical formulations comprise sufficient buffer to maintain pH shifts.

[0231] In certain embodiments, the bulking agent is selected from the group consisting of sucrose and trehalose (i.e., trehalose dihydrate). In certain embodiments, the bulking agent is sucrose. In certain embodiments, the bulking agent is trehalose. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25 mM to about 300 mM bulking agent. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM bulking agent. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 63 mM trehalose dihydrate.

[0232] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25 mg / mL SG, about 51 mM MES, about 63 mM trehalose dihydrate, and about 0.025% PS 80. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 25 mg / mL SG, about 51 mM MES, about 63 mM trehalose dihydrate, and about 0.025% PS 80. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 25 mg / mL SG, about 51 mM MES, about 63 mM trehalose dihydrate, and about 0.025% PS80.

[0233] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25 mg / mL SG, about 51.25 mM buffer, about 62.5 mM bulking agent, and about 0.025% surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 25 mg / mL SG, about 51.25 mM buffer, about 62.5 mM bulking agent, and about 0.025% surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 25 mg / mL SG, about 51.25 mM buffer, about 62.5 mM bulking agent, and about 0.025% surfactant, optionally wherein the pH is about 6.5.

[0234] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25 mg / mL SG, about 51.25 mM MES, about 62.5 mM trehalose dihydrate, and about 0.025% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 25 mg / mL SG, about 51.25 mM MES, about 62.5 mM trehalose dihydrate, and about 0.025% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 25 mg / mL SG, about 51.25 mM MES, about 62.5 mM trehalose dihydrate, and about 0.025% PS 80, optionally wherein the pH is about 6.5.

[0235] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 20 mg / mL SG, about 41 mM buffer, about 50 mM bulking agent, and about 0.02%surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 20 mg / mL SG, about 41 mM buffer, about 50 mM bulking agent, and about 0.02% surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 20 mg / mL SG, about 41 mM buffer, about 50 mM bulking agent, and about 0.02% surfactant, optionally wherein the pH is about 6.5.

[0236] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 20 mg / mL SG, about 41 mM MES, about 50 mM trehalose dihydrate, and about 0.02% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 20 mg / mL SG, about 41 mM MES, about 50 mM trehalose dihydrate, and about 0.02% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 20 mg / mL SG, about 41 mM MES, about 50 mM trehalose dihydrate, and about 0.02% PS 80, optionally wherein the pH is about 6.5.

[0237] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 30 mg / mL SG, about 61.5 mM buffer, about 75 mM bulking agent, and about 0.03% surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 30 mg / mL SG, about 61.5 mM buffer, about 75 mM bulking agent, and about 0.03% surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 30 mg / mL SG, about 61.5 mM buffer, about 75 mM bulking agent, and about 0.03% surfactant, optionally wherein the pH is about 6.5.

[0238] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 30 mg / mL SG, about 61.5 mM MES, about 75 mM trehalose dihydrate, and about 0.03% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 30 mg / mL SG, about 61.5 mM MES, about 75 mM trehalose dihydrate, and about 0.03% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 30 mg / mL SG, about 61.5 mM MES, about 75 mM trehalose dihydrate, and about 0.03% PS 80, optionally wherein the pH is about 6.5.

[0239] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM buffer, about 100 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM buffer, about 100 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM buffer, about 100 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.5.

[0240] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM MES, about 100 mM trehalose dihydrate, and about 0.04% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM MES, about 100 mM trehalose dihydrate, and about 0.04% PS 80, optionally wherein the pH is about 6.5. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM MES, about 100 mM trehalose dihydrate, and about 0.04% PS 80, optionally wherein the pH is about 6.5.

[0241] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 20 mg / mL to about 40 mg / mL SG, such as about 20 mg / mL to about 25 mg / mL, about 20 mg / mL to about 30 mg / mL, about 25 m / mL to about 40 m / mL, about 25 mg / mL to about 30 m / mL, or about 30 mg / mL to about 40 mg / mL.

[0242] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 20 mM to about 85 mM buffer, such as about 41 to about 82 nM, about 50 mM to about 82 mM, about 51 mM to about 82 mM, about 61 mM to about 82 mM, or about 62 mM to about 82 mM.

[0243] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 50 mM to about 200 mM bulking agent, such as about 50 nM to about 100 mM, about 50 mM to about 75 mM, about 62 mM to about 100 mM, about 63 mM to about 100 mM, about 100 mM to about 150 mM, or no more than 190 mM.

[0244] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 0.01%w / v surfactant to about 0.04%w / v surfactant, such as about 0.01%w / v to about 0.02%w / v, about 0.01%w / v to about 0.025%w / v, or about 0.01%w / v to about 0.03%w / v.

[0245] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM buffer, about 190 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM buffer, about 190 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM buffer, about 190 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.0.

[0246] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM buffer, about 190 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM buffer, about 190 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM buffer, about 190 mM bulking agent, and about 0.04% surfactant, optionally wherein the pH is about 6.0.

[0247] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0248] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0249] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0250] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certainembodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0251] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM phosphate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM phosphate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM phosphate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0252] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM phosphate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM phosphate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM phosphate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0253] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM maleate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM maleate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM maleate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0254] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 82 mM maleate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 82 mM maleate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 82 mM maleate, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0255] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8.

[0256] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8.

[0257] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.01% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.01% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM histidine, about 190 mM sucrose, and about 0.01% PS 80, optionally wherein the pH is about 5.8.

[0258] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM histidine, about 150 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM histidine, about 150 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM histidine, about 150 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 5.8. In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 20 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.02% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 20 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.02% PS 80, optionally wherein the pH is about 6.0. In certainembodiments, the pharmaceutical formulations as disclosed herein consist of about 20 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.02% PS 80, optionally wherein the pH is about 6.0.

[0259] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25.9 mg / mL SG, about 20 mM MES, about 126.4 mM sucrose, and about 0.026% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 25.9 mg / mL SG, about 20 mM MES, about 126.4 mM sucrose, and about 0.026% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 25.9 mg / mL SG, about 20 mM MES, about 126.4 mM sucrose, and about 0.026% PS 80, optionally wherein the pH is about 6.0.

[0260] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 25.9 mg / mL SG, about 20 mM MES, about 253.6 mM sucrose, and about 0.026% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 25.9 mg / mL SG, about 20 mM MES, about 253.6 mM sucrose, and about 0.026% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 25.9 mg / mL SG, about 20 mM MES, about 253.6 mM sucrose, and about 0.026% PS 80, optionally wherein the pH is about 6.0.

[0261] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM MES, about 100 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM MES, about 100 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM MES, about 100 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0262] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0263] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 40 mg / mL SG, about 20 mM MES, about 280 mM sucrose, and about 0.04% PS 80,optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 40 mg / mL SG, about 20 mM MES, about 280 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 40 mg / mL SG, about 20 mM MES, about 280 mM sucrose, and about 0.04% PS 80, optionally wherein the pH is about 6.0.

[0264] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 54.1 mg / mL SG, about 20 mM MES, about 126.4 mM sucrose, and about 0.054% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 54.1 mg / mL SG, about 20 mM MES, about 126.4 mM sucrose, and about 0.054% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 54.1 mg / mL SG, about 20 mM MES, about 126.4 mM sucrose, and about 0.054% PS 80, optionally wherein the pH is about 6.0.

[0265] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 54.1 mg / mL SG, about 20 mM MES, about 253.6 mM sucrose, and about 0.054% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 54.1 mg / mL SG, about 20 mM MES, about 253.6 mM sucrose, and about 0.054% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 54.1 mg / mL SG, about 20 mM MES, about 253.6 mM sucrose, and about 0.054% PS 80, optionally wherein the pH is about 6.0.

[0266] In certain embodiments, the pharmaceutical formulations as disclosed herein comprise about 60 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.06% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist essentially of about 60 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.06% PS 80, optionally wherein the pH is about 6.0. In certain embodiments, the pharmaceutical formulations as disclosed herein consist of about 60 mg / mL SG, about 20 mM MES, about 190 mM sucrose, and about 0.06% PS 80, optionally wherein the pH is about 6.0.

[0267] In certain embodiments, the pharmaceutical formulations disclosed herein have a pH of about 5.5 to about 7.0; about 6.0 to about 7.0; or about 6.2 to about 6.8. In certain embodiments, the pharmaceutical formulations disclosed herein of a pH of about 5.5; 5.6; 5.7; 5.8; 5.9; 6.0; 6.1; 6.2; 6.3; 6.4; 6.5; 6.6; 6.7; 6.8; 6.9; or 7.0. In certain embodiments, the pharmaceutical formulations disclosed herein have a pH of about 6.5. In certain embodiments, the pharmaceutical formulations have a pH of 6.0. In certain embodiments, the pharmaceutical formulations have a pH from about 5.8 to about 6.5. In certain embodiments, the pharmaceutical formulations have apH from about 6.0 to about 6.5. In certain embodiments, the pharmaceutical formulations have a buffer that maintains pH of the formulation at about 5.8 to about 6.5; at about 5.8 to about 6.0; at about 6.0 to about 6.5; at about 6.0, and / or at about 6.5. In certain embodiments, the pharmaceutical formulation comprises sufficient buffer to maintain pH of the formulation during lyophilization.

[0268] In certain embodiments, the pharmaceutical formulations disclosed herein have a pH of5.5 to 7.0; 6.0 to 7.0; or 6.2 to 6.8. In certain embodiments, the pharmaceutical formulations disclosed herein of a pH of 5.5; 5.6; 5.7; 5.8; 5.9; 6.0; 6.1; 6.2; 6.3; 6.4; 6.5; 6.6; 6.7; 6.8; 6.9; or 7.0. In certain embodiments, the pharmaceutical formulations disclosed herein have a pH of 6.5.In certain embodiments, the pharmaceutical formulations have a pH of 6.0. In certain embodiments, the pharmaceutical formulations have a pH from 5.8 to 6.5. In certain embodiments, the pharmaceutical formulations have a pH from 6.0 to 6.5. In certain embodiments, the pharmaceutical formulations have a buffer that maintains pH of the formulation at 5.8 to 6.5; at 5.8 to 6.0; at 6.0 to 6.5; at 6.0, and / or at 6.5. In certain embodiments, the pharmaceutical formulation comprises sufficient buffer to maintain pH of the formulation during lyophilization.

[0269] In certain embodiments, the pharmaceutical formulations as disclosed herein are stable. In certain embodiments, the pharmaceutical formulations disclosed herein have about 10 mg / mL to about 70 mg / mL SG and are stable. In certain embodiments, the pharmaceutical formulations disclosed herein have about 15 mg / mL to about 70 mg / mL SG and are stable.

[0270] In certain embodiments, the pharmaceutical formulations (i.e., the liquid or liquid prelyophilization formulations) disclosed herein are stable for at least about 30 days. In certain embodiments, the pharmaceutical formulations (i.e., the liquid or liquid pre-lyophilization formulations) disclosed herein are stable for at least 30 days in accelerated or stressed conditions. In certain embodiments, the pharmaceutical formulations (i.e., the liquid or liquid pre- lyophilization formulations) for at least 18 months at < -30°C or < -35°C. In certain embodiments, the pharmaceutical formulations (i.e., the liquid or liquid pre-lyophilization formulations) for 36 months at < -30°C or < -35°C. In certain embodiments, the pharmaceutical formulations maintain free SN-38 at < 1.0 pg / mg; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 0.4 pg / mg; < 0.3 pg / mg; < 0.2 pg / mg; or < 0.1 pg / mg for at least 18 months or for 36 months.

[0271] In certain embodiments, the pharmaceutical formulations maintain DAR at > 6.0; > 6.1;> 6.2; > 6.3; > 6.4; > 6.5; > 6.6; > 6.7; > 6.8; > 6.9; > 7.0; > 7.1; > 7.2; > 7.3; > 7.4; > 7.5; > 7.6;> 7.7; > 7.8; > 7.9; or > 8.0 for at least 18 months or for 36 months.

[0272] In certain embodiments, and as shown, for example, in FIGS. 27A-27D, the pharmaceutical formulations disclosed herein are stable for about at least about 30 days at atemperature of -20°C. In certain embodiments, the pharmaceutical formulations disclosed herein have a stable DAR for at least about 30 days at a temperature of -20°C. In certain embodiments, the pharmaceutical formulations as disclosed herein have a which DAR is about 7.4 or greater after storage at -20°C for at least about 30 days. In certain embodiments, the pharmaceutical formulations as disclosed herein have a % free SN-38 of about 2.2 or less after storage at -20°C for at least about 30 days. In certain embodiments, the pharmaceutical formulations as disclosed herein are stable for at least about 30 days at a temperature of -20°C irrespective of the concentration of SG in the formulation. In certain embodiments, the pharmaceutical formulations as disclosed herein have a stable DAR irrespective of the concentration of SG in the formulation.

[0273] In certain embodiments, and as shown, for example, in FIGS. 23A-23D, the pharmaceutical formulations (i.e., the liquid or liquid pre-lyophilization formulations) as disclosed herein are stable for at least about 30 to 60 hours at 5°C, 12°C, 18°C and 25°C. In certain embodiments, the pharmaceutical formulations as disclosed herein have a DAR which is about 7.3 or greater after storage for at least about 30 to 60 hours at 5°C.

[0274] In certain embodiments, the pre-lyophilization liquid formulations as disclosed herein have < 2.0%; < 2.1%; < 2.2%; < 2.3%; < 2.4%; < 2.5%; < 2.6%; < 2.7%; < 2.8%; < 2.9%; < 3.0%;< 3.1%; < 3.2%; < 3.3%; < 3.4%; or < 3.5% HMW species at release. In certain embodiments, the pre-lyophilization liquid formulations have < 3.0% HMW species at release. In certain embodiments, the pre-lyophilization liquid formulations have < 3.2% HMW species on stability for 18 to 36 months.

[0275] In certain embodiments, the pre-lyophilization liquid formulations as disclosed herein at release have < about 1.0 pg / mg; < about 0.9 pg / mg; < about 0.8 pg / mg; < about 0.7 pg / mg; < about 0.6 pg / mg; < about 0.5 pg / mg; < about 0.4 pg / mg; < about 0.3 pg / mg; < about 0.2 pg / mg;< about 0.1 pg / mg FDRI at release of the drug substances as disclosed herein. In certain embodiments, the pre-lyophilization liquid formulations have FDRI at the LOQ.

[0276] In certain embodiments, the pre-lyophilization liquid formulations as disclosed herein at release have 1.0 pg / mg; 0.9 pg / mg; 0.8 pg / mg; 0.7 pg / mg; 0.6 pg / mg; 0.5 pg / mg; 0.4 pg / mg; 0.3 pg / mg; 0.2 pg / mg; 0.1 pg / mg FDRI at release.

[0277] In certain embodiments, the pre-lyophilization liquid formulations as disclosed herein at release have a FDRI of less than 0.05 pg / mL, which is the limit of quantification (LOQ).

[0278] In certain embodiments, the pre-lyophilization liquid formulations has disclosed herein have a viscosity of about 1.1 to about 1.4 at 20°C. In certain embodiments, SG concentration may be increased while maintaining viscosity in this range.

[0279] The pre-lyophilization liquid formulations as disclosed herein are intended to be lyophilized. The lyophilized drug product is subsequently intended to be reconstituted to a targetSG concentration of 10 mg / mL. Stability must be maintained during the process of converting the pre-lyophilization liquid formulation (drug substance) to the lyophilized formulation (drug product), the formulation must maintain stability. Many factors may affect stability. An amount (volume) of pre-lyophilization liquid formulation is filled into each vial. The volume of liquid formulation itself filled into each vial may affect stability. Additionally, the choice of stopper on the vial may affect stability, i.e., as it may affect head space in the vial. Furthermore, free-thaw cycles during the lyophilization process may affect stability.

[0280] In certain embodiments, the pre-lyophilization liquid formulation is stable for 18 to 36 months at < -35°C. In certain embodiments, the pre-lyophilization formulation is stable for 36 months at < -35°C.Lyophilized or Reconstituted Pharmaceutical Formulations Comprising SG (Drug Product)

[0281] In certain embodiments, disclosed herein are lyophilized pharmaceutical formulations comprising (when reconstituted) about 10 mg / mL to about 70 mg / mL SG. In certain embodiments, disclosed herein are lyophilized pharmaceutical formulations comprising about 10 mg / mL to about 70 mg / mL SG. In certain embodiments, the lyophilized pharmaceutical formulations comprise about 10 mg / mL to about 70 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the lyophilized pharmaceutical formulations disclosed herein comprise about 40 mg / mL SG (when reconstituted). In certain embodiments, the lyophilized pharmaceutical formulations disclosed herein comprise about 10 mg / mL SG (when reconstituted).

[0282] In certain embodiments, disclosed herein are lyophilized pharmaceutical formulations comprising (when reconstituted) 10 mg / mL to 70 mg / mL SG. In certain embodiments, disclosed herein are lyophilized pharmaceutical formulations comprising 10 mg / mL to 70 mg / mL SG. In certain embodiments, the lyophilized pharmaceutical formulations comprise 10 mg / mL to 70 mg / mL SG, a surfactant, and a buffer. In certain embodiments, the lyophilized pharmaceutical formulations disclosed herein comprise 40 mg / mL SG (when reconstituted). In certain embodiments, the lyophilized pharmaceutical formulations disclosed herein comprise 10 mg / mL SG (when reconstituted).

[0283] In certain embodiments, the lyophilized pharmaceutical formulations comprise a bulking agent. In certain embodiments, the bulking agent is selected from the group consisting of sucrose, trehalose, and mannitol. In certain embodiments, the bulking agent is sucrose. In certain embodiments, the bulking agent is trehalose. In certain embodiments, the buffer is selected from the group consisting of MES, histidine, phosphate, and maleate. In certain embodiments, the buffer is selected from the group consisting of MES and histidine. In certain embodiments, the buffer is MES. In certain embodiments, the buffer is histidine. In certain embodiments, the buffer is not succinate. In certain embodiments, the buffer is not citrate. In certain embodiments, thebuffer is not succinate or citrate. In certain embodiments, the buffer is a combination of two or more buffers.

[0284] In certain embodiments, the lyophilized formulations as disclosed herein are stable at intended, accelerated, and stressed conditions. In certain embodiments, the lyophilized formulations are stable for the shelf life of the drug product. In certain embodiments, the lyophilized formulation is stable for at least 36 months at 2-8°C.

[0285] In certain embodiments, and as shown in FIGS. 29A-29D, the lyophilized pharmaceutical formulations as disclosed herein are stable for at least about 30 days at a temperature of 40°C (accelerated or stressed conditions). In certain embodiments, the DAR of the lyophilized pharmaceutical formulations as disclosed herein is about 7.4 or greater after storage at 40°C for at least about 30 days. In certain embodiments, % free SN-38 released from the lyophilized formulations as disclosed herein is less than about 3.5 after storage for at least about 30 days at 40°C.

[0286] In certain embodiments, the stability of the lyophilized pharmaceutical formulations as disclosed herein are dependent on the ratio of bulking agent (i.e., sucrose) concentration to protein (i.e., SG) concentration. For example, in certain embodiments, the % HMW species increases during storage, and the extent of the increase depends on the ratio of bulking agent concentration to SG concentration.

[0287] In certain embodiments, the pharmaceutical formulations disclosed herein are stable for at least about 30 days both as a pre-lyophilized liquid pharmaceutical formulation at -20°C and as a lyophilized pharmaceutical formulation at 40°C. For example, in certain embodiments, if a particular pharmaceutical formulation is stable as a pre-lyophilized formulation, that same formulation is stable as a lyophilized formulation. Stability as both a pre-lyophilized liquid formulation and as a lyophilized formulation is important for pharmaceutical products that are, for example, configured for intravenous use.

[0288] In certain embodiments, the lyophilized formulations disclosed herein comprise about 180 mg to about 200 mg SG. In certain embodiments, the lyophilized formulations comprise about 180 mg SG. In certain embodiments, the lyophilized formulations comprise about 200 mg SG. In certain embodiments, the lyophilized formulations disclosed herein are configured for bulk production. In certain embodiments, the lyophilized formulations disclosed herein are stored in 30R vials.

[0289] In certain embodiments, the lyophilized formulations as disclosed herein have < 2.0%; < 2.1%; < 2.2%; < 2.3%; < 2.4%; < 2.5%; < 2.6%; < 2.7%; < 2.8%; < 2.9%; < 3.0%; < 3.1%; < 3.2%; < 3.3%; < 3.4%; < 3.5%; < 3.6%; < 3.7%; < 3.8%; < 3.9%; or < 4.0% HMW species at release and / or during intended, accelerated and / or stressed storage conditions and / or during theshelf life. In certain embodiments, the lyophilized formulations have < 2.0%; < 2.1%; < 2.2%; < 2.3%; < 2.4%; < 2.5%; < 2.6%; < 2.7%; < 2.8%; < 2.9%; < 3.0%; < 3.1%; < 3.2%; < 3.3%; < 3.4%; < 3.5%; < 3.6%; < 3.7%; < 3.8%; < 3.9%; or < 4.0% HMW species for the intended shelf life of the drug product, i.e., 24 or 36 months at 2-8°C. In certain embodiments, the lyophilized formulations have < 3.5% HMW species at release. In certain embodiments, the lyophilized formulations have < 4.4 % HMW species on stability.

[0290] In certain embodiments, the lyophilized formulations as disclosed herein have < 1.0 pg / mg free SN-38; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 0.4 pg / mg; < 0.3 pg / mg; or < 0.2 pg / mg free SN-38 at release and / or during intended, accelerated, and / or stressed storage conditions. In certain embodiments, the lyophilized formulations as disclosed herein have < 1.0 pg / mg free SN-38; < 0.9 pg / mg; < 0.8 pg / mg; < 0.7 pg / mg; < 0.6 pg / mg; < 0.5 pg / mg; < 0.4 pg / mg; < 0.3 pg / mg; or < 0.2 pg / mg free SN-38 for the intended shelf life of the drug product, i.e., 24 or 36 months at 2-8°C.

[0291] In certain embodiments, the lyophilized formulations as disclosed herein have about 1.0 pg / mg free SN-38; about 0.9 pg / mg; about 0.8 pg / mg; about 0.7 pg / mg; about 0.6 pg / mg; about 0.5 pg / mg; about 0.6 pg / mg; about 0.5 pg / mg; about 0.4 pg / mg; about 0.3 pg / mg; or about 0.2 pg / mg free SN-38 at release and / or during intended, accelerated, and / or stressed conditions. In certain embodiments, the lyophilized formulations as disclosed herein have about 1.0 pg / mg free SN-38; about 0.9 pg / mg; about 0.8 pg / mg; about 0.7 pg / mg; about 0.6 pg / mg; about 0.5 pg / mg; about 0.6 pg / mg; about 0.5 pg / mg; about 0.4 pg / mg; about 0.3 pg / mg; or about 0.2 pg / mg free SN-38 for the intended shelf life of the drug product, i.e., 24 or 36 months at 2-8°C.

[0292] In certain embodiments, the lyophilized formulations as disclosed herein have a moisture or water content. In certain embodiments, the moisture or water content is < 1 % w / w. In certain embodiments, the moisture or water content is about <1% w / w at release or TO.

[0293] In certain embodiments, the lyophilized formulations as disclosed herein have a moisture or water content. In certain embodiments, the moisture or water content is about 1 % w / w. In certain embodiments, the moisture or water content is about 1% w / w at release or TO.

[0294] In certain embodiments, the lyophilized formulations disclosed herein are obtainable or obtained by lyophilizing an aqueous formulation comprising SG, wherein the aqueous formulation comprises a buffer that maintains pH during lyophilization at about 5.8 to about 6.0; or at 5.8 or 6.0. These lyophilized formulations are stable.

[0295] In certain embodiments, the lyophilized formulations disclosed herein are obtainable from or obtained by lyophilizing an aqueous formulation of SG, wherein the aqueous formulation comprises about 15 mg / mL to about 70 mg / mL SG. These lyophilized formulations are stable.

[0296] In certain embodiments, the lyophilized formulations disclosed herein are obtainable from or obtained by lyophilizing an aqueous formulation of SG, wherein the aqueous formulation comprises 15 mg / mL to 70 mg / mL SG. These lyophilized formulations are stable.

[0297] In certain embodiments, disclosed herein are stable lyophilized formulations of SG, wherein the formulation is made by lyophilizing an aqueous solution comprising 15 mg / mL to 70 mg / mL SG. In certain embodiments, the lyophilized formulations are obtainable from or obtained by lyophilizing a pre-lyophilization liquid formulation comprising 10 mg / mL - 40 mg / mL SG. In certain embodiments, the lyophilized formulations are obtainable from or obtained by lyophilizing a pre-lyophilized liquid formulation comprising 25 mg / mL SG.

[0298] Disclosed herein are methods and process suitable for producing SG in bulk. For example, in certain embodiments, vial size is reduced from 50R to 30R to enable production of more vials per run. In certain embodiments, the pre-lyophilization liquid formulation has a higher concentration of SG. This permits a lower volume (“fill volume”) of drug substance being added to the vial, which in turn permits a smaller vial size. A smaller vial size permits more vial loading capacity per run. In certain embodiments, for example, to have about 180 mg to about 200 mg of SG per vial, if a starting formulation of 10 mg / mL of SG drug substance is used, the target fill volume of each vial is about 20 mL (for each vial to have about 180 mg to about 200 mg of SG). If, however, a starting formulation of 25 mg / mL of SG drug substance is used, the target fill volume for each vial is about 8 mL (for each vial to have about 180 mg to about 200 mg of SG). Lower target fill volumes shorten the lyophilization cycle, thus enabling faster, more efficient production. It is to be understood, however, that increased protein concentration (i.e., increased SG) in the drug substance may pose increased stability issues.

[0299] In certain embodiments, the pharmaceutical batches, formulations, and compositions disclosed herein are produced by running the batches, formulations, and compositions through at least two different filtration methods. In certain embodiments, the pharmaceutical batches, formulations, and compositions disclosed herein are produced by running the batches, formulations, and compositions through two different filtration methods, one being absorptive filtration and the other one being non-ab sorptive filtration.

[0300] In certain embodiments, disclosed herein are pharmaceutical formulations comprising SG and having a DAR of about 6.7 to about 8.0. In certain embodiments, the pharmaceutical formulations have a DAR of about 6.7 to about 7.8. In certain embodiments, the pharmaceutical formulations comprising SG have a DAR of about 6.7 to about 7.8 as measured by size-exclusion ultraperformance liquid chromatography (SE-UPLC) with ultraviolet (UV) detection at 280 nm and 366 nm as disclosed herein.

[0301] Accurate measurement of DAR for a therapeutic biologic is very important for its therapeutic efficacy and tolerability. mAb formulations

[0302] In certain embodiments, disclosed herein are mAb formulations. In certain embodiments, the mAb formulations are stable. These mAb formulations may be used to produce the drug substance as disclosed herein. These mAb formulations may comprise about 10 g / L - 60 g / L hRS7. In these formulations, the mAb is hRS7. In certain embodiments, the mAb formulations comprise 50 mg / mL to 60 mg / mL hRS7 and are stable. In certain embodiments, the mAb formulations comprise 50 mg / mL to 60 mg / mL hRS7 and sodium acetate. In certain embodiments, the hRS7 formulations comprise about 0.01M sodium acetate and have a pH of about 5.7. In certain embodiments, the hRS7 formulations are stable and comprise 60 g / L hRS7 and sodium acetate. In certain embodiments, the hRS7 formulations are stable and comprise 50 g / L hRS7 and sodium acetate.

[0303] In certain embodiments, the mAb formulations are stable for 24-36 months at 2-8°C. In certain embodiments, the mAb formulations are stable for 36 months at < -30°C.

[0304] In certain embodiments, the mAb formulations as disclosed herein have < 1.0%; < 1.1%; < 1.2%; < 1.3%; < 1.4%; < 1.5%; < 1.6%; < 1.7%; < 1.8%; < 1.9%; < 2.0%; < 2.1%; < 2.2%; < 2.3%; < 2.4%; < 2.5%; < 2.6%; < 2.7%; < 2.8%; < 2.9%; < 3.0%; < 3.1%; < 3.2%; < 3.3%; < 3.4%; or < 3.5% HMW species at mAb release. In certain embodiments, the mAb formulations have <1.2% HMW species at mAb release. In certain embodiments, the mAb formulations have <1.5% HMW species on stability.

[0305] In certain embodiments, the mAb (i.e., hRS7) formulation is stable while stored at -30°C for at least three years. In certain embodiments, the mAb (i.e., hRS7) formulation is stable while stored at 2-8°C for up to three years.

[0306] Storage

[0307] In certain embodiments, and as shown in Table 47, the storage for the mAb, drug substance, and drug product are as follows:

[0308] Table 47.Methods

[0309] In certain embodiments, disclosed herein are methods to improve the stability of SG. Exemplary methods include formulating SG at a pH of 5.8 to 6.0 rather than at 6.5, thereby improving the stability of SG. In certain embodiments, disclosed herein are methods to improve the stability of SG drug substance and / or SG drug product comprising adjusting the pH of the drug substance and / or the drug product to 5.8 to 6.0; thereby improving the stability.

[0310] In certain embodiments, disclosed herein are methods of improving the stability of a high concentration hRS7 formulation, comprising formulating hRS7 with sodium acetate, thereby improving the stability of a high concentration hRS7 formulation.

[0311] In certain embodiments, disclosed herein are methods of reducing free drug-related impurities in SG, such as SG drug substance, comprising filtering crude ADC through an activated carbon filter, thereby reducing free drug-related impurities.

[0312] In certain embodiments, disclosed herein are methods of reducing % HMW species in an SG drug substance or an SG drug product, comprising conjugating SG at a higher concentration of hRS7. In certain embodiments, the method comprises using an hRS7 concentration of greater than 6 mg / mL hRS7 in the conjugation mixture.

[0313] In certain embodiments, disclosed herein are methods of improving the stability of SG drug substance formulations comprising greater than 10 mg / mL SG, the methods comprising protecting the drug substance from light.

[0314] In certain embodiments, disclosed herein are methods of treating unresectable locally advanced or metastatic triple-negative breast cancer in a human patient who has received two or more prior systemic therapies, at least one of them for metastatic disease, the method comprising administering to the patient in need thereof an SG drug product obtained from, or obtainable from, an SG drug substance comprising about 15 mg / mL to about 40 mg / mL SG.

[0315] In certain embodiments, disclosed herein are methods of treating unresectable locally advanced or metastatic hormone receptor (HR)-positive, HER2-negative (IHC 0, IHC 1+, or IHC 2+ / ISH-) breast cancer in a human patient who has received endocrine-based therapy and at least two additional systemic therapies in the metastatic setting comprising administering to the patient in need thereof an SG drug product obtained from, or obtainable from, an SG drug substance comprising about 15 mg / mL to about 40 mg / mL SG.EXAMPLESEXAMPLE 1Method of Manufacture of SG Drug SubstanceSummary

[0316] Example 1 describes the process that was used to manufacture the drug substance for sacituzumab govitecan (SG) (“Process 2”). A summary of the process is shown in FIG. 5. The process comprises reduction of the mAb (hRS7 IgGl K), conjugation of the mAb (hRS7) with the drug linker (CL2A-SN38) to produce a crude ADC, carbon depth filtration of the crude ADC, ultrafiltration / diafiltration (UF / DF) of the carbon-filtered ADC, and formulation of the filtered ADC to produce a drug substance comprising 25 mg / mL SG. The mAb and conjugate are protected from light during the manufacturing process.

[0317] The process in this Example was a continuous process with minimal to no hold times between process steps. hRS7 IgGlK and CL2A-SN38 were purified prior to conjugation. The conjugation procedure involved generation of free thiols on hRS7 IgGlK and subsequent mal eimide conjugation with CL2A-SN-38 (conjugation via a thiol-mal eimide bond). The conjugation process yielded a crude conjugated product (“crude ADC”) at a concentration of about 30 mg / ml. The crude conjugated product was purified using carbon depth filtration to remove process-related impurities including excess drug-linker. The purified conjugated product (carbon- filtered ADC) was then buffer exchanged via ultrafiltration / diafiltration (UF / DF) prior to formulation. The buffer-exchanged product was diluted with concentrated conditioning additives to reach the final drug substance protein concentration (25 mg / mL) and buffer composition (about 51 mM MES, about 62.5 mM trehalose, about 0.025% PS80, pH 6.5). The product was processed with an aseptic filtration and packaged to afford the final drug substance. The mAb (hRS7) and the ADC were protected from light. mAb Preparation and Reduction

[0318] Reduction was performed on hRS7 IgGlk (“mAb”) pooled from one or more lots and filtered through a sterilizing grade PES filter. After pooling, but prior to reduction, the mAb solution was conditioned with dilution buffer to achieve a target pH and buffer matrix. The mAb solution was then reduced with 5.5 equivalents of tris(2-carboxyethyl)phosphine (TCEP-HCL) for 240 min at 15-25 °C. mAb Input (mAb pooled solution)

[0319] The mAb input was a solution (pooled from multiple lots) of 50 mg / mL or 60 mg / mL mAb in 10 mM sodium acetate at a pH of 5.7, as shown in Tables 2A and 2B. The mAb input solution was filtered through a sterilization graded polyethersulfone (PES) filter (0.2 pM PES). The mAb input tank temperature target was 20°C, and the temperature range was 15°C-25°C.mAb pH Adjustment (mAb Conditioning)

[0320] Prior to reduction, the mAb input solution was conditioned with a dilution buffer (“mAb Conditioning Solution”) to achieve a target pH and buffer matrix. The mAb Conditioning Solution, as shown in Table 2A, (55 mM MES, 11 mM EDTA, pH 6.6) was added to the tank at a volume of 14.0 mL / g mAb. The resulting mAb solution was mixed for at least 15 minutes with the mAb Conditioning Solution. The pH after this step (mAb conditioning) was pH 6.3 - 6.7.

[0321] As an alternate approach to this step, the mAb Conditioning Solution as shown in Table 2B (71.7 mM MES, 14.3 EDTA, pH 6.5) was added to the tank at a volume of 14.0 mL / g mAb. The pH after this step (mAb conditioning) was pH 6.3 - 6.7. mAb Solution Reduction

[0322] The mAb solution was thereafter reduced with a reduction solution, as shown in Table 2A. (“mAb Reduction Reaction Solution”) [(5.5 molar equivalents of tris(2- carboxyethyl)phosphine (TCEP-HCL) (0.04 M TCEP in WFI)] for 240 min at 15-25 °C. The pH was adjusted to the target pH of 6.3. The pH for end of the reduction was pH 6.2-6.5. The mAb solution at the end of the reduction step was referred to as “Reduced mAb Pool.”

[0323] As an alternate approach to this step, the mAb solution was thereafter reduced with a reduction solution, as shown in Table 2B (“mAb Reduction Reaction Solution”) for 240 min at 15-25°C. The pH was adjusted to the target of 6.2. The pH for the end of reduction was 6.0-6.4. The post reduction titration solution was eliminated from this approach.Table 2A - mAb Solutions Prior to ConjugationTable 2B - Other mAb Solutions Prior to Conjugation

[0324] As shown in FIG. 1, the minimum time and temperature required to reach a robust reduction was determined. The minimum temperature was determined to be 15°C, and the minimum time was determined to be 4 hours. Temperatures below 15°C were determined to impact DAR at a target reduction time of 4 hours. Times below 4 hours outside of the target temperature were found to adversely affect DAR. Process criteria was determined to specify minimum of 4 hours for reduction and tank temperature at 20°C.Titration

[0325] Post-reduction, the Reduced mAb Pool was titrated with a “Post Reduction Titration Solution” (250 mM MES, pH 6.5).Conjugation Reaction (Conjugation of hRS7 to CL2A-SN38 drug-linker)

[0326] The conjugation was performed on the Reduced mAb Pool. The Reduced mAb Pool had a concentration of 30 mg / mL, and 10 molar equivalents of CL2A-SN-38 (DL) were added at a pH of 6.3. The DL was weighed and dissolved in dimethyl sulfoxide (DMSO) to prepare a 50 mM DL solution. The DL solution was added over ~5 minutes to ensure solution temperature did not exceed 25°C or 26°C as the addition was exothermic. A supplementary addition of DMSO was made to rinse the addition line and to adjust the total DMSO concentration of the reaction to 5% (w / v). The conjugation reaction was allowed to proceed at room temperature for a target of 20 minutes. Conjugation mixing time start was defined after DMSO rinse was added. Conjugation mixing end time was defined as the start of post-conjugation sampling. After conjugation was completed, temperature was reduced to 15°C. Carbon depth filtration may continue prior to reaching target temperature.Conjugation Process Parameters

[0327] The conjugation reaction operation temperature was 20°C, with a target range of 17°C- 25°C. The CL2A-SN-38 volume was 1.35 mL / g mAb, with a target range of 1.32-1.38 mL / g mAb. The CL2A-SN-38 solution was 50 mM CL2A-SN-38 in DMSO. The conjugation mixing time was 20 minutes, with a target range of 10-30 minutes. The operating temperature postconjugation was 15°C, with a target range of 4°C-25°C. The product of this step was a “crude conjugate,” or a “crude ADC”. After conjugation, the crude conjugate was held at 15°C for 12 hours.

[0328] As an alternate approach to the conjugation process parameters delineated above, the conjugation reaction operation temperature was 20°C, with a target range of 14-26°C. The CL2A- SN-38 volume was 1.35 mL / g mAb, with a target range of 1.32-1.38 mL / g mAb. The CL2A-SN- 38 solution was 50-75 mM CL2A-SN-38 in DMSO. The conjugation mixing time was 20 minutes, with a target range of 8-60 minutes. The operating temperature post-conjugation was15°C, with a target range of 4°C-25°C. The product of this step was a “crude conjugate,” or a “crude ADC”. After conjugation, the crude conjugate was held at 15°C for 12 hours.Quench

[0329] Quenching after conjugation was not required.Carbon Depth Filtration

[0330] Carbon depth filtration was the first purification step following the conjugation. The crude conjugate was passed through a single stage depth filter, which removed excess, unconjugated CL2A-SN-38 linker. In an alternate approach, the crude conjugate was passed through two stages of CR40 depth filters. The pool was collected after filtration through a sterilizing grade filter. The free drug-related impurities were measured pre-carbon depth filtration in the crude conjugate, and again post-carbon-depth filtration.

[0331] The carbon depth filter was processed in the following manner:

[0332] Step 1 : Pre-Use Carbon Depth Filter WFI (filter wetting solution) Flush: The carbon filter was flushed with a filter wetting solution. The flush volume target was 200 L / m2, with a target range of > 100 L / m2. The flux (LMH) target was 600. The depth filter inlet pressure (psi) target range was < 30.

[0333] Step 2: Pre-Use Carbon Depth Filter Equilibrium: The carbon filter was equilibrated with a filter equilibration solution: 20 mM histidine, pH 5.7. The equilibrium volume target range was > 20 L / m2. The flux (LMH) target was 600. The depth filter inlet pressure (psi) target range was < 30.

[0334] Sept 3 : Product Carbon Depth Filtration: The target range for loading density (of diluted, crude antibody-drug conjugate) was < 2250 g / m2or <5000 g / m2(total allowable was < 10,000 g / m2) and flux (LMH) target was 100. The depth filter inlet pressure (psi) target range was < 30. Thereafter, the pool was collected after filtration through a sterilizing grade filter. The target range for loading density (of the pool) was < 5000 g / m2.

[0335] Step 4: Product Recovery Chase: The sterilizing grade filter was flushed with 20 mM histidine, pH 5.7. The target flush volume range was < 30 L / m2. The flux (LMH) target was 100. The depth filter inlet pressure (psi) target range was < 30.

[0336] Step 5 Pool Hold Short Term: The pool tank was held at a target temperature of 15-25 °C for < 12 hours (hold time was determined from the isolation of the pool tank to the start of the ultrafiltration / diafiltration UF1 phase). Pool Hold Long Term: The pool tank was held at a target temperature of 2-8°C for < 48 hours (hold time was determined from the isolation of the pool tank to the start of the ultrafiltration / diafiltration UF1 phase).UF / DF

[0337] Ultrafiltration / Diafiltration (UF / DF) was a tangential flow filtration process used to both concentrate and buffer exchange the carbon filtered pool. In this step, the carbon-filtered ADC is retained by a membrane filter (Pall Corporation, Omega™ membrane) while smaller species, specifically process related impurities, pass through the membrane pores, and are removed. The specific tangential flow filtration (TFF) cassettes used in this unit operation were 50kDa cutoff modified PES membranes.

[0338] The load material was carbon-filtered ADC, diluted to 20 mg / mL. The operating temperature was 2 FC (range 4-26°C). The membrane was loaded at 800 g / m2(target range 500- 1100 g / m2).

[0339] The sanitization solution was 0.5 M sodium hydroxide, the equilibration / diafiltration / product recovery flush solutions were 25 mM MES pH 6.3. For ultrafiltration 1, the feed flow rate (LLM - liters per minute per total square meters of membrane area) was 4.5, and the target concentration was 20 g / L (range 12 g / L - 27 g / L). For diafiltration, the flow rate (LMM) was 4.5, and the number of diafiltration volumes used was 9. For ultrafiltration 2, the flow rate (LMM) was 4.5, and the target concentration was 50 g / L.Conditioning and Drug Substance Filtration

[0340] The UFDF pool was conditioned and formulated into the drug substance with a target SG concentration of 25 mg / mL. The recovered UFDF pool was brought to 5°C and was diluted and then adjusted with a concentrated conditioning solution to achieve the target final drug substance composition. The volumes of diluent and conditioning solution required were determined based on the expected final bulk volume at 25 g / L. After dilution, the bulk material was conditioned to reach the target ADC and excipient concentrations with 4X conditioning buffer (129 mM MES, 250 mM trehalose, 0.1% PS80 at pH 6.5). After conditioning, the formulated drug substance was filtered by a 0.22 pm filter into polycarbonate bottles in preparation for freezing and storage. The drug substance filled bottles were protected from light and subjected to a low temperature freezing process and then transferred to long-term frozen storage conditions.

[0341] As shown in Table 26, carbon filtration can remove free drug-related impurities across multiple scales. This table shows the FDRI pre- and post-carbon filtration.Table 26.EXAMPLE 2Quantitation of Free SN-38 in SG (DS and DP) by Reverse Phase HPLC

[0342] Free SN-38 can be quantified as described below.

[0343] Protein-related content in the SG sample was removed with a protein precipitation reagent and unconjugated SN-38 (a.k.a. free SN-38) was extracted while SN-38 conjugated to antibody was removed along with the antibody. SN-38 was separated on a reverse phase HPLC column [(Waters™ X-Select CSH Cl 8 3.5 pm, 3.0 x 100 mm column (P / N: 186005262)] isostatically using an acidified mobile phase to ensure detection of a single isomer. The flowrate was 0.600 mL / min. Mobile Phase A (MPA) was 0.2% trifluoroacetic acid (TFA) in water. Mobile Phase B (MPB) was 100% acetonitrile. The injection volume was 5 pL and autosampler temperature was 5 + / - 3 °C.

[0344] SN-38 was quantitated against a SN-38 standard using fluorescence detection (FLD). FLD settings were excitation at 373 nm, emission at 540 nm. The run time was 17 minutes.

[0345] The gradient program was as described in Table 3.Table 3.EXAMPLE 3UV-Size Exclusion UPLC Analysis of Purity and Drug to Antibody Ratio (DAR) on SG Drug Substance and Drug Product

[0346] This analytical method to determine purity and DAR was based on the observed resolution of protein peaks using size-exclusion ultraperformance liquid chromatography (SE- UPLC) with ultraviolet (UV) detection at 280 and 366 nm. This method was also used for relative quantification of the high molecular weight (HMW) peak group. The UPLC system usedwas Waters ACQUITY™ H-Class bio-inert UPLC System. A 5mm path length flow cell was required for analysis (e.g., Waters ACQUITY™ PDA 5 mm titanium flow cell, P / N 205000613).Standard and Sample Preparation

[0347] Reference Standard Preparation. One vial of SG reference standard (RS) material was reconstituted with 20 mL of purified water. The storage of reconstituted 10 mg / mL SG RS at ambient temperature was limited to a maximum of 6 hours. The reconstituted RS was used within 72 hours when stored at 5+ / -3°C. An aliquot to be tested is transferred into an HPLC vial.

[0348] Test Sample (TS) Preparation (Drug Product). One vial of SG was reconstituted with 20 mL of purified water. An aliquot to be tested was transferred into an HPLC vial.

[0349] Test Sample (TS) Preparation (Bulk Drug Substance)

[0350] A vial of bulk drug substance material was thawed at 5+ / - 3°C until analysis. At the time of analysis, an aliquot to be tested as transferred into an HPLC vial. The storage of thawed TS at ambient conditions was limited to a maximum of 6 hours. The TS was used within 72 hours after initial thaw, when stored at 5 ± 3°C. Chromatography conditions are shown in Table 4.Table 4.Column installation, flush, and equilibration

[0351] The new column was flushed with purified water for a minimum of 65 minutes using the instrument settings in Table 4. Following the purified water flush, the column was equilibrated with buffered mobile phase for a minimum of 145 minutes. An exemplary injection sequence is shown in Table 5.Table 5.EXAMPLE 4SG Drug Substance Formulations and Drug Product Formulations

[0352] Various formulations of the SG drug substance were prepared and tested on stability. These formulations were tested to be stable for time appropriate for their intended use, i.e., sufficient stability to be stored prior to the lyophilization step. These formulations were intended to be of sufficient stability to produce a drug product that ultimately would be administered via intravenous infusion. These formulations were lyophilized to produce a drug product having about 180 mg SG to about 200 mg SG.

[0353] Table 6 shows exemplary formulations of SG drug substance. These are liquid and prelyophilization formulations.Table 6.

[0354] In this example, (3) drug substance formulations (IX, 2X, and 2.5X) were tested on stability.

[0355] These formulations were generated by conjugating hRS7 mAb to yield hRS7-CL2A-SN- 38 (SG) ADC at 2.5X strength and further diluting to 2X and IX formulations with water. Prior to conjugation, the source mAb was 10 mg / mL hRS7 in PBS, reprocessed into 60 mg / mL hRS7 in 10 mM acetate, pH 5.5. The SG drug substance was produced by conjugating hRS7 mAb to yield 40 g of ADC material, purified by ultrafiltration-diafiltration (UF / DF), then formulated to 2.5X strength.Stability Study Plans

[0356] Stability studies were conducted for SG drug substance (pre-lyophilization), drug product (lyophilized), and saline (reconstituted). Table 7 describes the stability study plan for the IX, 2X, and 2.5X drug substance formulations.Table 7.

[0357] In Table 7, the number “1” refers to “1” vial. (12) vials were tested for each drug substance formulation (IX, 2X, and 2.5X).

[0358] The drug product (lyophilized formulation) was generated for IX, 2X, and 2.5X formulations by filling 200 mg of SG per vial using differing amounts of drug substance formulation [(IX, 20 mL in 50R vials), (2X, 10 mL in 30R vials), and (2.5X, 8 mL in 30R vials)]. Lyophilization was performed for all three formulations simultaneously in one run, using the cycle parameters (PD: -10°C / 150 mTorr) and as shown in Table 8. The total lyophilization cycle time was about 5 days. Once lyophilized, each of the vials had the same amount of SG (a different volume was used to achieve the same amount SG), i.e., about 180 mg or about 200 mg of SG. “Lyo” refers to “lyophilization.”Table 8.

[0359] The lyophilized drug product was tested for stability as indicated in Table 9. The drug product was stored for the time indicated, but a small amount was reconstituted immediately prior to stability testing.Table 9.

[0360] Numbers refer to the number of vials tested. (22) vials were tested for each formulation (IX, 2X, and 2.5X).

[0361] Reconstituted drug product (saline) was tested for stability. The lyophilized samples were reconstituted in saline, stored for up to three days, and tested on stability as shown in Table 10.Table 10.

[0362] Numbers refer to the number of vials tested. (4) vials were tested for each formulation (IX, 2X, and 2.5X).Results of Stability Testing

[0363] Table 11 shows pH at different temperatures and time points for the IX, 2X, and 2.5X drug substance formulations. Unless otherwise stated, under the “Time” column, the numbers refer to months.Table 11.Table 12 shows protein concentration (SG) at different temperatures and time points for the IX, 2X, and 2.5X drug substance formulations. Unless otherwise stated, under the “Time” column, the numbers refer to months.Table 12.

[0364] Table 13 shows purity / aggregation at different temperatures and time points for the IX, 2X, and 2.5X drug substance formulations. FIGS. 6A-6D show % HMW species by SEC [as described in Example 2] at different temperatures and time points for the IX, 2X, and 2.5X drug substance formulations.Table 13.SEC

[0365] Table 14 shows DAR at different temperatures and timepoints for the IX, 2X, and 2.5X drug substance formulations. FIGS. 7A-7D show DAR by SEC (as described in Example 3) at different temperatures and time points for the IX, 2X, and 2.5X drug substance formulations.Table 14.

[0366] Table 15 shows pH at different temperatures and timepoints for the IX, 2X, and 2.5X drug product (lyophilized) formulations.Table 15.

[0367] Table 16 shows purity / aggregation at different temperatures and time points for the IX, 2X, and 2.5X drug product formulations. FIGS. 8A-8E show % HMW species by SEC-DAR, % main peak, and % LMW species by SEC at different temperatures and timepoints for the IX, 2X, and 2.5X drug product formulations. FIG. 8 A shows % HMW species by SEC-DAR at 25°C for the IX, 2X, and 2.5X drug product formulations. FIG. 8B shows % HMW species by SEC-DAR at 5°C for the IX, 2X, and 2.5X drug product formulations. FIG. 8C shows % HMW species by SEC-DAR at 40°C for the IX, 2X, and 2.5X drug product formulations. FIG. 8D shows % main peak by SEC at 25°C and 40°C for the IX, 2X, and 2.5X drug product formulations. FIG. 8E shows % LMW species by SEC at 25°C and 40°C for the IX, 2X, and 2.5X drug product formulations.Table 16.

[0368] Table 17 shows DAR at different temperatures and time points for the IX, 2X, and 2.5X drug product formulations. FIGS. 9A-9C show DAR by SEC-DAR at different temperatures and timepoints for the IX, 2X, and 2.5X drug product formulations.Table 17.

[0369] Table 19 shows % acidic species at different temperatures and time points for the IX, 2X, and 2.5X drug product formulations. FIGS. 10A-10B show % acidic and % basic species by CEX at different temperatures and timepoints for the IX, 2X, and 2.5 drug product formulations.Table 19.EXAMPLE 5

[0370] The drug substance formulations in Example 5 were prepared as shown in FIG. 16. The following was the Stability Study Plan for this Example: (1) Liquid Stability: 5°C for 1, 2, 3, and 7 days and (2) Liquid Stability: 25°C for 1, 2, and 3 days.

[0371] To produce the various formulations, lyophilized drug product in a 50R vial was reconstituted. Samples were stored as liquid formulations and not lyophilized formulations.

[0372] Table 20 shows the drug substance formulations that were tested in this Example.Table 20.

[0373] The drug substance formulations were prepared by reconstituting 50R vials of lyophilized drug product with WFI [4.7 - 20 mL, depending on final drug substance formulation],

[0374] FIGS. 17A-17B show the % HMW species at various timepoints for the IX, 2X, 3X, and 4X drug substance formulations at 5°C (FIG. 17A) and at 25°C (FIG. 17B). FIG. 17C shows % HMW species at various freeze-thaw cycles (at -80°C and 5°C) for the IX, 2X, 3X, and 4X drug substance formulations. FIGS. 18A-18C show DAR at various timepoints or freeze thaw cycles for the IX, 2X, 3X, and 4X drug substance formulations. FIGS. 19A-19C show % free SN-38 at various timepoints or freeze thaw cycles for the IX, 2X, 3X, and 4X drug substance formulations. As shown in FIG. 19 A, from TO to T15 (0 days to 15 days), the least change in DAR occurred in the formulation with the highest SG concentration, the 4X formulation. Higher formulation concentration correlates with increased aggregation, but also decreased drug-linker cleavage (more stable DAR, less % free SN-38 increase).EXAMPLE 6Buffer Study

[0375] Six buffers were evaluated on liquid (drug substance) and lyophilized (drug product) SG stability. The following was fixed: 40 mg / mL SG; 190 mM sucrose (bulking agent or filler); 0.04% PS80, at pH 6.0. The formulations are described in Table 21. Note that the formulation numbers in Table 21 refer to this Example 6 only. Buffer concentrations were 20mM or 82 mM.The samples were prepared as shown in FIG. 11. Formulations Fl to F12 were prepared by reconstituting lyophilized drug product with 10 mL water, pooling, filtering, dialysis, and reformulating with different buffers. F13 was prepared by reconstituting lyophilized drug product with 4.7 mL water, pooling, and lyophilizing. The source material was TRODELVY® drug product (lyophilized) in a 50R vial.Table 21.

[0376] Formulations F3-F6 (having succinate or citrate buffers) crashed out during the buffer exchange / re-formulation and were thus excluded from the stability studies.

[0377] FIGS. 12A and 12B show % HMW species and % free SN-38 (366 nm) at a temperature of 18°C and at different timepoints (T=0 to T=50 hours) of the different pre-lyophilization drug substance formulations.

[0378] As shown in FIG. 12A, Lots MOI 1 and MOI 1-N (10 mg / mL SG; 21 mM MES; 25 mM trehalose; and 0.01 w / v % PS80), Fl, and F2 had less % HMW species at TO, and the percentage either decreased over time or remained steady. As shown in FIG. 12B, Fl 2, Fl 1, and F10 had less % free SN-38 at TO, and the percentage increased less rapidly over time. Compared to the other formulations, Fl and F13 had the highest % free SN-38 at T=0, and the percentage increased more rapidly over time. Regardless, the % free SN-38 in these formulations did not increase as rapidly as did the % free SN-38 in Lots MOI 1 and MOI 1-N.

[0379] FIGS. 13A and 13B show % HMW species and % free SN-38 of the different prelyophilization drug substance formulations at a temperature of 5°C and at different timepoints. As shown in FIG. 13 A, Fl, and F2 had less % HMW species at TO, and the percentage decreased over time. As shown in FIG. 13B, F12, Fl 1, and F10 had less % free SN-38 at TO, and the percentage increased less rapidly over time.

[0380] FIGS. 14A and 14B show % HMW species and % free SN-38 at -20°C and -80°C of the different pre-lyophilization drug substance formulations at timepoints of T=0 and T=30 days. For each formulation in these figures, the first bar represents TO; the second bar represents T30 at - 20°C; and the third bar represents T30 at -80°C. As shown in FIGS. 14A and 14B, all formulations were stable at both temperarures for about 30 days.

[0381] FIGS. 15A and 15B show % free SN-38 by SEC and free SN-38 by RP-HPLC as described herein of the different drug substance formulations at 5°C at T=0 to T=15 days. In FIG. 15 A, the cirle represents Fl; square represents F2; triangle represents F7; upside-down triangle represents F8; diamond represents F9; larger circle represents F10; larger square represents Fl 1; larger triangle represents F12; and larger upside down triangle represents F13. In FIG. 15B, the circle represents Fl; square represents F2; triangle represents F7; “x” represents F8; and the star represents F13.

[0382] As shown in FIGS. 15A and 15B, Fl and F13 had the highest free SN-38, and F8 had the lowest.

[0383] Lyophilized drug product was tested for stability (formulations were reconsituted immediately prior to stability testing). FIGS. 20A and 20B show % HMW species and % free SN-38 at 40°C from T=0 to about T=45 for the drug product formulations. F14 (control) had the following formulation: 10 mg / mL SG; 22 mM MES; 22 mM trehalose dihydrate; 0.01% PS80; pH 6.5. Referring to FIG. 20A, at T 40 - 50: lowest to highest % HMW species: (bottom line to top line - F2; Fl; F8; F7; F9; Fl 1; F10; F14 (control); F13; and F12 (top line). Referring to FIG. 20B, the bottom line at T 40 - T 50 represents Fl l and the the line right on top of this line represents F10. The two lines at top at T 40 - T 50 represent F 13 and F14 (control - next-to-top line).

[0384] As shown in FIG. 20A, drug product formulations with MES or histidine exhibited 2.5- 5-fold slower aggregation than F14 (control).

[0385] FIGS. 21A-21B show % HMW species of the drug product formulations with MES or histidine at various timepoints at 40°C. Referring to FIG. 21 A, at TO - T 10, referring from bottom line to top line: F2, Fl; F7; F8; F14 (control); and F13. Referring to FIG. 21B, at TO - T 10, referring from bottom line to top line: F2; Fl; F7; and F8.

[0386] Table 22 shows the slope analysis from T=0 to T=30 of the graphs in FIGS. 21A-21B.As shown in Table 22, F14 had the largest slope (and thus the greatest aggregation over time).Table 22.

[0387] FIGS. 22A-22B show free SN-38 (pg / mL) (FIG. 22 A) and % free SN-38 (FIG. 22B) at 40°C for the drug product lyophilized formulations at various timepoints. Referring to FIG. 22A, at TO, from bottom line to top line: 50R control; F2 (square); F7 (diamond); Fl; and F13.EXAMPLE 7Sucrose-Protein StudyStudy Design

[0388] Eleven formulations (Table 23) were generated to investigate the effects of SG (10 mg / mL to 70 mg / mL) and sucrose (0 to 280 mM) concentration on drug substance (liquid) stability. Nine of those formulations (F01 to F09) were used to investigate the effects of SG concentration and sucrose concentration on drug product (lyophilized) stability.Table 23.

[0389] The stability study plan is described in Table 24.Table 24.

[0390] To produce the drug substance formulations used in this study, lyophilized drug product across several vials having about 200 mg SG each was reconstituted with 10 mL of purified water to produce about 20 mg / mL SG. During reconstitution, the vials were pooled and stored on ice until the final pool (about 1.7 L) was ready for buffer exchange with about 10L of 20 mM MES at pH 5.75 and concentrated to 69.9 mg / mL over about 6 hours. Appropriate volumes of SG, sucrose, and MES stock solutions were then mixed to generate each formulation (Table 23) and subsequently filtered through 0.22pm membranes. After filtration, aliquots of each formulation were frozen at -20°C for stability studies. To create the lyophilized drug product formulations for this study, after filtration, the remaining material of each of the 9 formulations was filled into 5 sets of 50-mL centrifuge tubes and frozen at -80°C. The samples were lyophilized in 20R vials. Thus, each pre-lyophilized formulation was the same as each lyophilized formulation (in terms of SG concentration and excipient concentration). Immediately prior to conducting the stability study, however, a portion of the lyophilized formulation was reconstituted to test stability.Stability Results - Liquid Drug Substance Formulations

[0391] DAR and released (free) SN-38 (SE-UPLC run time extended and late eluting peaks integrated to capture free SN-38) was measured by SE-UPLC. Free SN-38 was measured by RP- UPLC.

[0392] FIGS. 23A-26D show the stability of liquid drug substance formulations as measured by HMW species %, monomer %, DAR, and % released (free) SN-38 (at 366 nm) at 5°C, 12°C, 18°C, and 25°C using SE-UPLC. The formulation labelled “S16M011” was a liquid drug substance formulation of 10 mg / mL SG; 25 mM trehalose; 20.5 mM MES; and 0.01% w / v PS80.

[0393] FIGS. 23A-23D show the stability of liquid drug substance formulations as measured by HMW species % (FIG. 23A), monomer % (FIG. 23B), DAR (FIG. 23C), and % released (free) SN-38 (at 366 nm) (FIG. 23D) at 5°C. Referring to FIG. 23 A, the top line (right-sided arrow) represents S16M011. The line below S16M011 was FOO. With respect to FIG. 23C, the line with the lowest starting DAR represents S16M011. With respect to FIG. 23D, S16M011 had the highest % released SN-38 at TO. HMW species % and monomer % were relatively constant duringthe storage period at 5°C. DAR value decreased slightly and % released SN-38 (measured by the relative peak area of SN-38 Peak 1 and Peak 2 of all SN-38 peaks at 366 nm) was in parallel with the reference (reconstituted S16M011 at 10 mg / mL). Change in DAR and % released SN-38 was not dependent on protein and sucrose concentrations.

[0394] FIGS. 24A-24D show the stability of liquid drug substance formulations as measured by HMW species % (FIG. 24 A), monomer % (FIG. 24B), DAR (FIG. 24C), and % released SN-38 (at 366 nm) (FIG. 24D) at 12°C. Referring to FIG. 24A, S16M011 had the highest % HMW species (top line). FOO had the second-highest % HMW species (line just below top line). Fl (bottom line) consistently had the lowest % HMW species. As shown in FIGS. 24C-24D, S16M011 (line represented by right-sided arrow) had the lowest DAR and highest % released SN- 38 at TO.

[0395] At 12°C, HMW species % and monomer % were relatively constant during the storage period and observed to be SG-concentration dependent. DAR and % released SN-38 did not appear to be dependent on the concentration of SG in the formulation.

[0396] FIGS. 25A-25D show the stability of liquid drug substance formulations as measured by HMW species % (FIG. 25 A), monomer % (FIG. 25B), DAR (FIG. 25C), and % released SN-38 (at 366 nm) (FIG. 25D) at 18°C. At 18°C, HMW species % and monomer % content of the formulations having an SG concentration of 60 mg / mL or above were relatively constant during the storage period. For formulations having an SG concentration of about 54 mg / mL or lower, a decreasing trend in HMW species % and an increasing trend in monomer % were observed. The DAR value for all the sample formulations decreased at a rate slightly less than the reference (S16M011) and the rate of increase of % released SN-38 for all formulations was slightly less than that of the reference (S16M011). Referring to FIG. 25A, S16M011 had the highest % HMW species (top line); FOO (next highest); and F9 (next highest). Referring to FIGS. 25C-25D, S16M011 had the lowest DAR and highest released free SN-38 at TO (represented by right-sided arrow).

[0397] FIGS. 26A-26D show the stability of liquid drug substance formulations as measured by HMW species % (FIG. 26 A), monomer % (FIG. 26B), DAR (FIG. 26C), and % released SN-38 (at 366 nm) (FIG. 26D) at 25°C. At 25°C, for all sucrose-containing formulations, HMW species % showed a decreasing trend during storage, and monomer % showed a slight increasing trend. HMW species % and monomer % appeared to be SG-concentration dependent. DAR for all formulations decreased at a rate comparable to that of SI6M011. The rate of % released SN-38 for all the formulations was less than that of S16M011. Change in DAR and % released SN-38 did not appear to be dependent on SG concentration and / or sucrose concentration. Referring to FIGS. 26A and 26C, the formulations represented by circle (FOO); right-sided arrow (S16M011);and left-sided arrow (S16M011) had the worst % HMW species and worst DAR. At all storage temperatures, F0 (without sucrose) had the highest HMW species % and lowest monomer %. Change in DAR did was not SG concentration dependent.

[0398] FIGS. 27A-27D shows the stability of the liquid drug substance formulations as measured by HMW species % (FIG. 27 A), monomer % (FIG. 27B), DAR (FIG. 27C), and % released SN-38 (at 366 nm) (FIG. 27D) at -20°C following storage for 30 days.Stability Results - Lyophilized Drug Product Formulations

[0399] Table 25 shows the lyophilization cycles with various primary drying temperatures employed for each set of samples.Table 25

[0400] FIGS. 28A-28D show % HMW species (FIG. 28 A), monomer % (FIG. 28B), % DAR (FIG. 28C) and % released free SN-38 (FIG. 28D) of the lyophilized drug product formulations (identified by sucrose concentration to protein concentration ratio) at various primary drying temperatures. The samples were reconstituted immediately prior to stability testing. A higher sucrose concentration to SG concentration ratio was correlated with a lower % HMW species content and a higher monomer % content. The primary drying temperatures at -15°C to 0°C had comparable impact on HMW and monomer content, however, primary drying temperature at 5°C resulted in a higher level of aggregation and lower level of monomer content than the other temperatures. Neither the sucrose to SG concentration ratio nor the primary drying temperature had a significant impact on DAR. A slight overall decreasing trend was observed for releasedSN-38 at a higher ratio of sucrose concentration to SG concentration. With respect to FIG. 28 A, F8 had the lowest % HMW species. With respect to FIG. 28C, Fl had the highest DAR.

[0401] FIG. 29A shows % HMW species results for lyophilized drug product formulations stored at 40°C for up to about 30 days. % HMW species increased during storage, and the extent of increase was dependent on the ratio of sucrose concentration to SG concentration. For example, with respect to formulation 5, the increase of % HMW species in the lyophilized formulation after storage for about 30 days was 0.86%.

[0402] FIG. 29B shows monomer % results for the lyophilized drug product formulations stored at 40°C for up to about 30 days. Monomer % decreased during the storage period, and the extent of decrease was dependent on the ratio of sucrose concentration to SG concentration. For example, with respect to formulation 5, the decrease during the storage period of 30 days was 0.82%, which was much less than the rate of decrease of the reference.

[0403] FIG. 29C shows DAR results for the lyophilized drug product formulations stored at 40°C for up to about 30 days. No trend in DAR was observed.

[0404] FIG. 29D shows % released SN-38 for the lyophilized drug product formulations stored at 40°C for up to about 30 days.

[0405] FIG. 30 shows free SN-38 concentration measured with RP-HPLC method for the lyophilized drug product formulations stored at 40°C for up to about 30 days. The samples were reconstituted for the stability measurement at 30 days, and the SG concentration was about 20 mg / mL after reconstitution, and the data was normalized to an SG concentration of 10 mg / mL.EXAMPLE 8Measurement of free drug-related impurities.

[0406] This Example describes an analytical procedure to quantify free SN-38, drug linker CL2A-SN-38, and related species present in SG in-process samples using reverse phase HPLC and fluorescence detection (FLD).

[0407] Post conjugation, free CL2A-SN-38 was a minor free species in the retentate during the UF / DF. The major free species detected were CL2A-SN-38 with its reactive mal eimide either conjugated with TCEP, reduced, or reacted with primary amine of lysine from the linker to form a cyclic degradation product as shown in FIG. 33.

[0408] For CL2A-SN-38 quantitation, the fluorescence response was assumed to be comparable between CL2A-SN-38 and related species since the fluorescence signal arises from the SN-38 moiety. Accordingly, all species (besides SN-38) were quantitated against the CL2A-SN-38 standard.

[0409] Free SN-38 and its carboxylate were also present in-process samples as well as bulk drug substance upon final formulation. SN-38 has a substantially higher response by FLD whenhydrolyzed from the linker and was quantitated against an SN-38 standard. This method utilized a hold period to convert the carboxylate form of SN-38 into the lactone form of SN-38. SN-38 was quantitated against the lactone form of SN-38 in the calibration curve.

[0410] All peaks were integrated in the chromatogram, starting from free SN-38. TCEP-CL2A- SN-38, reduced CL2A-SN-38, cyclized CL2A-SN-38, and CL2A-SN-38 were identified and quantitated.EXAMPLE 9Impact of pH of hRS7-SN-38 at Protein Concentration of 20 mg / mL

[0411] In this study, five formulations containing 20 mg / mL SG in 20 mM MES were prepared over a target pH range from 5.25 to 6.50. Study samples were stored at 5°C, 25°C, or 40°C and periodically evaluated by appearance, protein concentration, turbidity, SE-UPLC, reducing CE- SDS, and CEX-HPLC methods after storage for 1 day and up to 30 days.

[0412] The results from the study show that less intensity of yellow color was observed at pH 6.0 or lower. Visible particles were also observed earlier at lower pH conditions after storage at 40°C than at other temperatures. The result for OD600 was also pH-dependent and lower pH showed more increment during storage. The SE-UPLC results showed that HMW species is pH- dependent and increased at a faster rate at a lower pH during storage. No pH-dependent results were observed for the LMW species.

[0413] DAR was pH-dependent, and the results showed that improved solution stability profile for SG by DAR was achieved by lowering the formulation pH from 6.5 to 6.0. A further decrease in pH from pH 6.0 to 5.3 did not improve DAR at a comparable magnitude as that from pH 6.5 to 6.0. A similar pH-dependent trend was observed for Fab main species measured by CEX-HPLC. Lower pH reduced the degree of yellow color in the SG solution. Under stress conditions, turbidity and particle formation were observed to worsen at lower pH; however, the differences between pH 6.5 and 6.0 were minimal.

[0414] Collectively, the data from this study illustrated that a formulation pH close to 6.0 was an optimal pH and provided an appreciable improvement in SG stability compared with the stability of the pH 6.5 formulation (i.e., a formulation having 10 mg / mL SG; 22 mM MES; 25 mM trehalose dihydrate; 0.01% PS80 at pH 6.5).

[0415] Specifically, five formulations containing 20 mg / mL SG in 20 mM MES were prepared over a pH range from 5.25 to 6.50 (Table 27). Study samples were stored at 2-8°C (i.e., 5°C), 22- 28°C (i.e., 25°C), and 40°C and periodically evaluated over a period of 30 days.Table 27. Target Sample MatrixTable 28. Stability ScheduleStudy Preparation

[0416] Target formulation buffer recipes were determined by titrating 20 mM solutions of MES and MES sodium salt over a broad pH range, as shown in Table 27. SG was sourced from drug product vials, each containing 200 mg of lyophilized material. For each of the five formulations, four vials were reconstituted using 5 mL of the target formulation buffer to achieve a concentration of about 40 mg / mL. Each set of four vials was then pooled and transferred to a dialysis cartridge for buffer exchange into the target formulation. All bulk solutions of about 20 mL were dialyzed with 2L of 20 mM MES at a time with a minimum of 3 changes over 41 hours.

[0417] Upon reaching the target formulation pH, the solutions were each adjusted to a concentration of 20 mg / mL SG with their corresponding target formulation buffer (Table 29A). The final concentrations and pH were recorded for each solution and are shown in Table 29B.Samples were then aseptically filled, under BSC into ready -to-use sterile glass vials, capped with stoppers (Table 29C), and crimped with metal seals. After filling, the vials were organized into boxes with lids to protect from light and placed at each storage condition.Table 29A.Table 29B.Table 29C.

[0418] FIGS. 45A-45C show protein concentration by Solo VPE over time.

[0419] Protein concentration was determined using the variable pathlength Solo VPE slopebased method (no sample dilution, absorbance at 280 nm, and an extinction coefficient of 1.6). As shown in FIGS. 45A-45C, there were no significant changes observed in protein concentrationacross formulations and over time and different temperatures 5°C (FIG. 45A); 25°C (FIG. 45B); 40°C (FIG. 45C).Turbidity by Plate ReaderThe UV-Vis spectra for SG showed absorbance of various structural components and yellow color (assumed to be associated with absorbances through 450 nm). To avoid potential interference at lower wavelengths, solution turbidity was measured at 600 nm with a plate reader. Optical density was determined by averaging two readings of two neat 200 pL replicates for each sample solution (blanked against corresponding buffer matrix) in a UV-transparent 96- well plate (Thermo, P / N 8404). OD 600 nm values ranged between 0.003 and 0.526 across all study samples.

[0420] FIG. 46 shows the absorbance spectrum of SG drug substance.

[0421] FIGS. 47A-C show the effect of pH on turbidity at 5°C (FIG. 47A); 25°C (FIG. 47B); and 40°C (FIG. 47C). The initial turbidity values were comparable across all formulations. Turbidity increased as the storage duration increased.

[0422] At 5 °C, changes in turbidity were not pH-dependent for up to 7 days. After storage for 14 and 30 days, a trend of higher turbidity was observed with decreasing pH.

[0423] At 25°C, turbidity increased relatively faster than at 5°C. After storage for 1 day and longer, a trend was again observed with higher turbidity correlating to decreased pH.

[0424] At 40°C, the turbidity increased much faster than at 25°C.Size Variant Distribution and D AR, by SE-UPLC

[0425] The size variant distribution and DAR of test samples was determined by size exclusion ultra-performance liquid chromatography (SE-UPLC). Both data sets were derived from single injections of each test sample. All samples were centrifuged to remove particles from the solution. Samples were injected neat and at a fixed volume to load on the column.HMW species - The effect of formulation pH on % HMW of SG is shown in FIG. 48A (5°C); FIG. 48B (25°C); and FIG. 48C (40°C).

[0426] Initial HMW species were in the range of 3.30% to 3.70% and found to be higher with increasing pH. Over storage at 5°C, 25°C, and 40°C, a pH-dependent decreasing trend for HMW species was observed for formulations with pH ranging from 5.29 to 5.95 (the lower the pH, the faster the decreasing trend). At pH 6.56, there is no significant decreasing trend over storage at 5°C, 25°C, or 40°C. Specific numerical values are shown in Table 30 below.Table 30.DAR by SE-UPLC

[0427] DAR ratios were interpolated from a linear formula based on the ratio of total integrated areas (366nm: 280nm) for each sample. This information was collected and leveraged from the same sample inj ections used for size variant analysis. The effect of formulation pH on DAR values is shown in FIGS. 49A (5°C); FIG. 49B (25°C); and FIG. 49C (40°C). Initial DAR values were very similar across formulations (7.19-7.20). As shown in FIG. 49A, referring to the left-side of the graph (i.e., at the lowest pH); the top line represents Day 0; second from the top represents Day 7; 3rdfrom the top represents Day 14; forth from the top represents Day 2; and bottom represents Day 30. As shown in 52B, referring to the left-side of the graph (i.e., at the lowest pH); the top line represents Day 0; second from the top represents Day 1; third from the top represents Day 2; fourth from the top represents Day 5; fifth from the top represents Day 7, and the bottom represents Day 14. Referring to FIG. 49C, and specifically to the left-side of the graph (i.e., at the lowest pH); the top line represents Day 0; second from the top represents Day 1; third from the top represents Day 2; and bottom represents Day 5.

[0428] During storage at 5°C, 25°C, and 40°C, DAR values at pH 6.0 or lower decreased slightly slower than those at pH 6.5.

[0429] Specific numerical values are shown in Table 31 below.Table 31.*DAR calculations were derived by the following formula:DAR = ( 14.53 x R) + 0.008, where R = (Protein Area 366nm / Protein Area 280nm) rounded to three significant digits.

[0430] In this study, the stability of SG at pH in the range from 5.29 to 6.56 at approximately 0.25-unit intervals were evaluated with 20 mM MES. The results show that pH 6.0-6.5 improves clarity and minimizes particle formation and optical density at OD600, while pH 5.3 -6.0 decreases HMW species of SE-HPLC and reduce release of SN-38 as measured by DAR.The data from this study indicated that pH 6.0 was an optimal pH and may provide an improvement in SG stability in compared with an SG DP with a pH of 6.5.EXAMPLE 10

[0431] Based on the foregoing studies, the following four formulations (as shown in Table 32) were selected for further development. It is to be understood that the formula identifiers, i.e., “Fl” and the like as used in this Example only refer to the formulations in this Example 10.The tables and Figures in this Example 10 refer to “Tl” and “T2 ” This refers to the upstream process by which SG was made. For example, referring to FIGS. 55A-B; and FIG. 56B, T2 in Fl refers to SG made by the T2 process as identified in Table 33A, but formulated as 10 mg / mL in the Fl formulation. This is reference to the upstream production process for SG, as shown in Table 33A.Table 32.Table 33A.

[0432] FIGS. 34A-34C show at what part in the process confirmation studies were conducted. FIG. 34A shows end-to-end process stability; FIG. 34B shows in-use studies; and FIG. 34C shows lyophilization cycle assessment.

[0433] FIGS. 35A-35B show % BMW species (FIG. 35 A) and free SN-38 (FIG. 35B) at various timepoints during the end-to-end process conversion of drug substance to drug product. Referring to FIG. 35 A, Referring to FIGS. 35 A and 35B, for each formulation, the first bar (from left to right) represents post-formulation; the second bar represents post freeze / thaw; the third represents post 24 hr 5 °C hold; the fourth represents pre-lyophilization, and the fifth represents Lyo TO. Fl had a lower starting % HMW species due to differences in UF / DF processing conditions and a smaller scale.

[0434] FIGS. 51 A and 5 IB show subvisible particles in drug substance formulations as shown in Table 32 by HIAC on stability. Referring to FIGS. 51A-51B, for each formulation, each bar represents, from left to right: first bar represents TO; second bar represents 1 month at 5°C; and third bar represents 3 months at - 80°C. FIG. 51 A shows subvisible particles per mL at > 10 pm particles / mL and FIG. 5 IB shows subvisible particles per mL at > 25 pm particles / mL.

[0435] FIGS. 52A and 52B show subvisible particles in drug product formulations as shown in Table 32 by HIAC on stability. Referring to FIGS. 52A-52B, for each formulation, each bar represents, from left to right: TO; 3 months at 5°C; 3 months at 25°C; and 3 months at 40°C. FIG. 52A shows subvisible particles per mL at > 10 pm parti cles / mL and FIG. 52B shows subvisible particles per mL at > 25 pm particles / mL.

[0436] Additional studies were conducted to determine IV bag compatibility. 20 mL of 0.9% sodium chloride injection, USP, was added to each vial containing 180 mg to 200 mg SG. The resulting concentration of SG was lO mg / mL. SG was diluted to 1.1 - 3.4 mg / mL for IV infusion. The first additional study identified the appropriate amount of PS80 (0.01% vs 0.04%) in formulations 2 and 3 of Table 32. The second additional study compared formulation 2 with the following T1 formulation: 10 mg / mL SG; 22 mM MES; 25 mM trehalose dihydrate; 0.01% PS80 at pH 6.5.

[0437] FIGS. 53 A (target concentration in IV bag 1.1 mg / mL; PS800.0011%) and53B (target concentration in IV bag 3.4 mg / mL; PS80 0.04%0.0034%) show SVP levels(subvisible particles) of formula 2 (40 mg / mL protein in 20 mM His, 190 mM sucrose, 0.04% PS80 in pH 5.8 before and after infusion. For each SVP level, the bars represent, from left to right: TO; 5C overnight; before infusion and after infusion.

[0438] FIGS. 54A (target concentration in IV bag 1.1 mg / mL; PS800.0003%) andFIG. 54B (target concentration in IV bag 3.4 mg / mL; PS80 0.01%0.0009%) show SVP levels(subvisible particles) of formula 3 (40 mg / mL protein in 20 mM His, 190 mM sucrose, 0.01% PS80 in pH 5.8 before and after infusion. There were minimal differences in SVP post-infusion regardless of PS80 levels. The critical micelle concentration (CMC) of PS80 is 0.0017%. For each SVP level, the bars represent, from left to right: TO; 5°C overnight; before infusion and after infusion.

[0439] FIGS. 55A-55B show results of additional study 2 (formula 2 vs. T1 formulation - Table 33B). FIG. 55A shows SVP by HIAC and FIG. 55B shows % SN-38 by SEC. The F2 formulation was comparable to the T1.0 formulation with respect to both.

[0440] FIGS. 56A-56B show % HMW species by SEC after dilution with saline (FIG. 56A) and in use (FIG. 56B). Referring to FIGS. 56A-56B, the top line (from left to right) represents T1 in LOF; T2 in F2; and T2 in LOF.Table 33B.

[0441] Table 34 shows that primary drying times were improved using F2 formulations compare to T1.0 formulation. There was no significant benefit to drying time at 150 mM sucrose (5% solids) vs. 190 mM sucrose (6% solids). For both formulations, there was acceptable moisture (< 1%); acceptable reconstitution time (< 1 min.); and acceptable appearance.Table 34.

[0442] FIGS. 36A-36B show stability of the drug substance formulations at an intended storage temperature of -80°C. FIG. 36A shows % HMW species as a function of time, and FIG. 36B shows free SN-38 as a function of time. As shown, there were only minor differences in the stability of all four drug substance formulations at the intended storage temperature. “T1.0” as used in the figures are different lots of the following drug substance formulation: 10 mg / mL SG; 25 mM trehalose; 0.01 (% w / v) PS80; pH 6.5. There were only minor differences between Fl -F4 formulations and the T1 formulation.

[0443] FIGS. 37A-B show stability of the drug product formulations at an intended storage temperature of 5°C. FIG. 37A shows % HMW species as a function of time, and FIG. 37B shows free SN-38 as a function of time. As shown, there were only minor differences in the stability of all four drug product formulations at the intended storage temperature. “T1.0” as used in the figures are different lots of the following drug product formulation: 10 mg / mL SG; 25 mM Trehalose; 0.01 (% w / v) PS80; pH 6.5.

[0444] FIGS. 38A-38B show stability of the drug substance formulations under stressed conditions of 5°C. As shown, there were only minor differences in the stability of all four drugsubstance formulations under stressed conditions. “T1.0” as used in the figures are different lots of the following drug substance formulation: 10 mg / mL SG; 25 mM Trehalose; 0.01 (% w / v) PS80; pH 6.5.

[0445] FIGS. 39A-39B show stability of the drug product formulations under stressed conditions of 40°C. As shown, there were only minor differences in the stability of all four drug product formulations under stressed conditions. “T1.0” as used in the figures are different lots of the following drug product formulation: 10 mg / mL SG; 25 mM Trehalose; 0.01 (% w / v) PS80; pH 6.5.

[0446] Under both intended and stressed conditions, the F1-F4 formulations had same or better stability than the T1 formulations.

[0447] As disclosed herein, higher sugar content (i.e., sucrose) led to better stability than lower sucrose concentrations; however, generally all formulations Fl -F4 were stable under intended and stressed conditions for both drug substance and drug product formulations.

[0448] There were no major differences observed in any product quality attribute between all four formulations at the intended, accelerated, and stressed stability temperatures for up to 3 months.

[0449] Additionally, higher sucrose concentration (190 mM) showed reduced % HMW species compared to the lower sucrose concentration (150 mM) in the lyophilized drug product on stability.

[0450] Additionally, the PS80 concentration post-dilution in IV bag would be less than the CMC concentration of PS80 and thus there is a potential risk for increase in sub-visible particles and / or protein loss due to adsorption to surfaces. There was no benefit with the lyo cycle time decrease with the formulation having 150 nM sucrose. Prior studies indicate better stability at higher sucrose concentration.EXAMPLE 11.Residual Moisture Study

[0451] This example evaluated the impact of residual moisture content on the product quality of SG drug product produced according to Example 1 (Process 2). This DP was lyophilized as a 25 mg / mL ADC formulated in 51 mM MES, 62.5 mM trehalose, 0.025% w / v PS80 at pH 6.5. The drug product was reconstituted with sterile water for injection to provide 10 mg / mL SG in 20.4 mM MES, 25 mM trehalose, 0.01% w / v PS80 at pH 6.5.

[0452] In this example, stability data was presented for samples stored over a 12-month period at an intended storage temperature of 5°C+ / - 3°C; over 6 months at an accelerated storage temperature of 25°C+ / - 2°C (60% relative humidity+ / - 5%); and over 3 months at a stressedstorage temperature of 40°C + / - 2°C (75% relative humidity + / - 5%). The study configuration is shown in Table 35.

[0453] Based on the 12-month stability data at the intended storage of 2-8°C, SG drug product quality was minimally impacted with residual moisture content up to 3.7%. At an accelerated storage of up to 6 months, product quality was also minimally impacted with a residual moisture content up to 3.7%. At stressed storage, significant change in product quality (%HMW, free SN- 38, and % acidic species) was observed especially for samples with moisture content of > 3 % (Range 3) and was correlated with increasing water content and lower Tg values. Both Type 1 vial types (Schott and Ompi) showed comparable product quality on stability at intended long term; accelerated; and stressed temperatures.

[0454] SG drug substance (at 10 kg scale) was used as the source material for the drug product. Three different water levels were spiked into the lyophilized DP vials to give three ranges of water content i.e., Range 1 (1-2%), Range 2 (2-3%) and Range 3 (> 3%) along with two controls (no water spiked) in two different vial types at the target moisture content (<1%). For lyophilized DP at various % water content, physicochemical stability was determined at intended storage temperature of 5°C for 36 months, at an accelerated temperature of 25°C / 60% RH over a period of 12 months, and at 40°C / 75% relative humidity for 6 months.Table 35. Study ConfigurationDP vial for these ranges are 30R Qni i (see materials table for additional details)

[0455] Two 2L PC bottles of 1.6 L SG drug substance were pulled from -80°C frozen storage and thawed for approximately 72 hours at 5°C. Once thawed, the SG drug substance was homogenized by inverting the bottle 10 times. Thereafter, the drug substance was pooled into a 10L PC bottle and filtered using a Corning bottle-top 0.22 pm PES filter in a LAF hood. Samplepreparation was performed in a LAF hood where a total of two hundred and seventy-four 3 OR glass vials (50 Schott and 224 Ompi) were filled with 8 mL of the filtered DS while keeping the vials and DS on ice. The vials were partially stoppered and loaded into the lyophilizer at shelf set point of 5°C and lyophilized using a LyoStar 3 development lyophilizer under the conditions specified in Table 36. Lyophilized vials were stoppered, crimp sealed, labeled by vial type (Schott or Ompi) and stored at 5°C.Table 36. Conditions for Residual Moisture Study - Drug Product LyophilizationPreparation of Water-Spiked DP Vials for Stability

[0456] DP vials were removed from 5°C storage and equilibrated to room temperature for 30 minutes. A 18G needle was used to vent the vial to prevent backpressure during spiking. Spiking was performed by placing the DP vial horizontally on the balance and injecting water onto the side of the vial close to the neck using a 50 pL Hamilton syringe affixed with a 25G needle. Water was added to the neck of the vial until water weight matched target range, see Table 37 for details. This step was performed for Range 1-3 samples only. Each DP vial was held horizontally to ensure water did not come into contact with the lyo cake as the stopper was sealed with parafilm. All samples were incubated at 25°C for 48 hours while maintaining the vials in a horizontal orientation. Once incubation was complete, vials were stored upright at 5°C until stability set down (TO) was initiated.Preparation of Water-Spiked DP Vials for Stability Table 37.Water Content

[0457] Lyophilized cakes at all timepoints were assessed for % water content by Karl Fisher. FIGS. 40A-40C show the change in water content from the initial water content range at intended, accelerated, and stressed storage conditions.

[0458] As shown in FIG. 40A, at intended long-term storage at 5°C, minimal changes in water content were observed after 12 months storage except for Range 2 at 6-months and except for Range 3 at 6-months and 12-months where there was higher water content level measured. This was most likely due to inherent variability in each vial due to manual spiking of water level in the vials. As shown in FIG. 40B, at the accelerated storage condition of 25°C, a similar increase in water content was observed for all ranges. Ompi and Schott controls increased by 0.8% and 0.7% respectively; Range 1 increased by 0.8%; Range 2 increased by 0.7%; and Range 3 increased by 0.8% after 6 months storage. As shown in FIG. 40C, after 3 months storage at stressed 40°C temperature, a similar increase in water content was observed for all ranges. Both Ompi and Schott controls increased by 0.6%, Range 1 increased by 0.5%, Range 2 increased by 0.5% and Range 3 increased by 0.4%. pH and Protein Concentration

[0459] Select time points were measured for pH and concentration. The overall protein concentration results were within 10% of the initial measured protein concentration at all timepoints and storage conditions across all moisture ranges. The pH results were within 0.1 pH units of the initial pH at all timepoints and storage conditions across all moisture ranges. Refer to Table 38 for individual data sets.Table 38.TiiuepointDetermination of Purity and D AR by SE-UPLC

[0460] The percent relative areas for main peak and high molecular weight species were monitored as well as drug antibody ratios using size exclusion chromatography. As shown in FIG. 41A, at intended storage at 5°C, all the samples across all moisture ranges demonstrated comparable increase in % HMW species (0.3-0.5%) with corresponding decrease in main peak after 12-months of storage. As shown in FIG. 41B, after 6-months at accelerated 25°C storage, an increase in % HMW species was observed in the control samples of 1.9, and the water-spiked samples in Ranges 1-3 increased in range of 1.3 - 1.6%. As shown in FIG. 41C, after 3-months of storage at stressed 40°C, controls and Range 1-2 samples showed % HMW species increase in the range of 3.3 - 3.8%, and Range 3 showed a higher increase in % HMW species of 4.6%. A higher increase in % HMW species at 40°C in Range 3 (>3%) samples as compared to other residual moisture levels (controls, Range 1 and Range 2) samples was directly correlated with the lower Tg onset (50.8°C) measured for Range 3 samples.

[0461] The drug antibody ratio (DAR) results were in the range of 7.2 to 7.5 (within assay variability) for all samples at intended storage (5°C); accelerated storage (25°C), and stressed storage (40°C) conditions. Table 39 shows the data sets.Table 39.Free SN-38

[0462] The amount of the unconjugated drug SN-38 was measured by reversed phase high performance liquid chromatography (RP-HPLC) after separation from the antibody conjugated to SN-38 through a precipitation process. The results were first presented in pg / mL. The ratio of free SN-38 to protein concentration was then used to report the results in pg / mg. As shown in FIG. 42A, at intended storage of 5°C (up to 12-months) and accelerated storage of 25°C (up to 6 months), minimal changes were seen across different moisture content levels after 12 months storage. As shown in FIG. 42B, after 3 months storage at stressed conditions (40°C), an increase of 0.3-0.4 pg / mg was measured in samples with water content <3% (controls, Range 1, Range 2) and 0.7 pg / mg increase for Range 3 sample. A higher increase in free SN-38 at 40°C in Range 3 (>3%) samples as compared to other residual moisture levels (controls, Range 1 and Range 2) samples can be directly correlated with the lower Tg onset (50.8°C) measured for Range 3 samples. Table 40 shows the data sets.Table 40.Potency

[0463] Binding was a cell-based bioassay that measured for biological activity through the cell binding of SG conjugates to cell surface receptors. The binding was measured to be in the range from 91-108%..

[0464] This study evaluated the impact of increasing residual moisture content in SG (produced by the process described in Example 1) lyophilized DP cakes on the ADC critical quality attributes. The moisture (water content) ranges evaluated in this study are Control (<1%), Range 1 (1-2%), Range 2 (2-3%), and Range 3 (> 3%).

[0465] Based on the 12-months’ stability data at intended storage of 2-8°C, product quality was minimally impacted with residual moisture content up to 3.7%. At accelerated storage up to 6- months, similar change in product quality (% HMW species and water content) was measured with residual moisture content up to 3.7%. At stressed storage, significant change in product quality (% HMW species, free SN-38 and % acidic species) was observed especially for samples with moisture content of > 3% (Range 3) and was correlated with increasing water content and lower Tg values. Both the Type 1 vial types (Schott and Ompi) showed comparable product quality on stability at intended long-term, accelerated, and stressed temperatures.EXAMPLE 12.

[0466] Improved % HMW species stability trends for Process 2 (Example 1) drug product at accelerated and stressed conditions.SG drug product was produced according to Process 1 (Example 16) and Process 2 (Example 1). Drug product made by Process 1 (Example 16) is identified in the figure by circles, and drug product made by Process 2 (Example 1) is identified in the figure by squares. The arrows identify the line that refers to drug product made by Process 2 at manufacturing scale (10 kg).As shown in FIGS. 43A-43B, under accelerated (FIG. 43 A) and stressed (FIG. 43B) conditions, drug product made by Process 2 (Example 1) shows improved HMW stability.EXAMPLE 13.FDRI

[0467] FDRI was measured at release (TO) for drug substance as produced by Example 1 (Process 2) and as produced by Example 16 (Process 1).

[0468] As shown in FIG. 44, the drug substance produced by Example 1 comprised less FRDI than the drug substance produced by Example 16 (Process 1).EXAMPLE 14.StabilityStability of mAb, drug substance, and drug product at various time points and temperatures were assessed. Tables 41A-B show stability of mAb at 50 g / L.Table 41A. Reformulation of mAb in Example 16 (Process 1) to 50 mg / mL in sodium acetate - StabilityTable 4 IB. mAb (hRS7) formulated as in Example 1 - Stability

[0469] Table 42 shows stability of the drug substance (25 mg / mL SG) obtained from a batch size of 10 kg. The drug substance was produced according to Example 1.Table 42.

[0470] Table 43 shows stability of drug product (180 mg - 200 mg SG) at various time points and temperatures. The drug product was produced according to Example 1. The drug product was produced from a 10 kg batch.Table 43.EXAMPLE 15.Viscosity

[0471] Viscosity of IX, 2X, and 4X drug substance formulations are shown in Table 44 below.The drug substance was obtained from a batch size of 10 kg.Table 44.

[0472] Viscosity of various SG drug substances formulations are shown below in Table 45.Table 45. Response Surface Design.EXAMPLE 16Another Method of Manufacture of SG Drug Substance

[0473] This Example describes the manufacture of SG according to another manufacturing process, Process 1. An alternate process for the manufacture of SG is shown in FIG. 50.

[0474] A high-level process overview is described below.

[0475] Thawing, dispensing, and preparation of solutions: hRS7 IgGk bulk solution, stored at 2- 8°C, was brought to room temperature and dispensed. CL2A-SN-38, stored at -20°C, was brought to room temperature before dispensing.

[0476] Pooling and reduction: Multiple batches of hRS7 IgGk bulk solution were pooled, with 0.2 M ethylene diamene tetraacetic acid (EDTA) and 0.04 phosphate-buffered saline (PBS) added to the solution before reduction with 0.04 TCEP at room temperature.

[0477] pH adjustment: A solution of sodium phosphate was used to adjust the pH of the solution to pH 6.5.

[0478] Conjugation: A first aliquot of DMSO was added to the conjugation tank while stirring. Subsequently, the solution of CL2A-SN-38 in DMSO was added within 5 minutes, followed by a second aliquot of DMSO. The solution is stirred for 20 + / - 10 minutes.

[0479] Quenching: A 10 mg / mL NEM solution was added, and the solution was stirred for 10- 60 minutes, with a target of 35 minutes.

[0480] Purification, buffer exchange, and concentration: Purification consisted of the following steps: (1) First UF: SG was concentrated to approximately 10 mg / mL. (2) Diafiltration, 30 diavolumes of 0.025 MES (buffer exchange) were used to remove free SN-38; free CL2A-SN-38, reagents and solvents (DMSO, TCEP, NEM, Phosphate, NaCl). (3) Second UF: SG was concentrated to approximately 20 mg / mL.

[0481] Formulation: The target weight required for a final protein concentration of 10 mg / mL was determined based on the concentration of DS measured by UV spectrophotometry. While stirring at 80-100 rpm, solutions of 0.25 M trehalose; 1% w / v polysorbate 80, 0.022 M MES, and pH 6.5 were prepared.

[0482] Bottling and freezing: The final solution was transferred (through a pre-flushed 0.22 pm filter) from the tank to the bottles, which were stored at -80°C + / - 10°C.

[0483] The target drug substance formulation produced by this process was 10 mg / mL SG; 22 mM MES; 22 mM trehalose; 0.01% PS80; pH 6.5. The vial was filled with 20 mL of the drug substance, and the material was lyophilized. Each vial had 180 mg - 200 mg SG. The drug product was made by reconstitution. After reconstitution, the drug product has the same formulation as the drug substance (after reconstitution, SG is 10 mg / mL).EXAMPLE 17Drug Substance Testing In-Process Strategy.

[0484] The Example 17 describes the in-process testing strategy for a drug substance according to Example 1.Table 46.EXAMPLE 18.Stability

[0485] This Example compares the stability of the drug substance made by the process in Example 16 (3 lots) and the drug substance (3 lots) made by the process in Example 1. The process in Example 16 was at 3.3 kg scale, and the process in Example 1 was at 10 kg scale.

[0486] FIGS. 57-61B show the comparisons in stability. As shown in the figures, the filled-in shapes (triangle, diamond, and circle) refer to the three different lots of the drug substance made by Example 16. As shown in the figures, the empty shapes (triangle, diamond, and circle) refer to the three different lots of the drug substance made by Example 1.

[0487] FIG. 57 compares the degradation rate (as measured by free SN-38) of the drug substance produced by the process in Example 1 compared with the degradation rate of the drug substance produced by the process in Example 16 under the stressed conditions of 2-8°C.

[0488] Table 47 shows the slopes of the (6) lots tested.Table 47.

[0489] FIGS. 58A-58C show the SEC / DAR degradation rate for drug substance made by the process in Example 16 and the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. The process in Example 16 was at 3.3 kg scale, and the process in Example 1 was at 10 kg scale. FIG. 58 A shows % HMW species as a function of time, FIG. 58B shows % main peak as a function of time, and FIG. 58C shows DAR as a function of time. The numerical data is shown in Table 48.Table 48.

[0490] FIGS. 59A-59B show rCE (reduced capillary electrophoresis-SDS) degradation rate assessment for drug substance made by the process in Example 16 and for the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. FIG. 59A shows % LC (light chain) + % HC (heavy chain) as a function of time and FIG. 59B shows %HL (heavy-light dimer) as a function of time. The numerical data is shown in Table 49.Table 49.

[0491] FIGS. 60A-60C show CEX degradation rate assessment for drug substance made by the process in Example 16 and for the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. FIG. 60A shows % acidic species as a function of time, FIG. 60B shows % fab main as a function of time; and FIG. 60C shows % Fab basic as a function of time. The numerical data is shown in Table 50A-C.Table 50A.Table 50B.Table 50C.

[0492] FIGS. 61 A-B show potency degradation rate assessment for drug substance made by the process in Example 16 and for the drug substance made by the process in Example 1 under stressed conditions of 2-8°C. FIG. 61 A shows % potency-cell binding as a function of time and FIG. 61B shows % potency-cytotoxicity as a function of time.

[0493] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

Claims

CLAIMSWhat is claimed is:

1. A stable liquid pharmaceutical formulation comprising: about 15 mg / mL to about 70 mg / mL sacituzumab govitecan (SG); a surfactant; and a buffer, wherein pH of the formulation is about 5.5 to about 6.8.

2. The stable liquid pharmaceutical formulation of claim 1, comprising a bulking agent.

3. The stable liquid pharmaceutical formulation of claim 2, wherein the bulking agent is selected from the group consisting of sucrose, trehalose, and mannitol.

4. The stable liquid pharmaceutical formulation of claim 2 or 3, wherein the bulking agent is sucrose.

5. The stable liquid pharmaceutical formulation of any one of claims 1-4, wherein the buffer is selected from the group consisting of MES, histidine, phosphate, and maleate.

6. The stable liquid pharmaceutical formulation of any one of claims 1-5, wherein the buffer is selected from the group consisting of MES and histidine.

7. The stable liquid pharmaceutical formulation of any one of claims 1-6, wherein the buffer is MES.

8. The stable liquid pharmaceutical formulation of any one of claims 1-6, wherein the formulation is stable at -20°C after storage for at least about 30 days.

9. The stable liquid pharmaceutical formulation of any one of claims 1-8, wherein DAR is about 7.4 or greater after storage at -20°C for at least about 30 days.

10. The stable liquid pharmaceutical formulation of any one of claims 1-9, wherein % free SN-38 is less than about 2.2 after storage for at least about 30 days at -20°C.

11. The stable liquid pharmaceutical formulation of any one of claims 1-10, wherein % free SN-38 is stable for at least about 30 days at -20°C.

12. The stable liquid pharmaceutical formulation of any one of claims 1-11, wherein % HMW species is stable for at least about 30 days at -20°C.

13. The stable liquid pharmaceutical formulation of any one of claims 1-12, wherein the formulation is stable irrespective of SG concentration.

14. The stable liquid pharmaceutical formulation of any one of claims 1-13, wherein DAR is about 7.3 or greater after storage at 5°C for about 60 hours.

15. The stable liquid pharmaceutical formulation of any one of claims 1-14, wherein DAR is stable for at least about 30 days as both a liquid formulation at -20°C and as a lyophilized formulation at 40°C (greater than 7.38).

16. A stable lyophilized pharmaceutical formulation comprising: about 10 mg / mL to about 70 mg / mL SG; a surfactant; and a buffer, wherein pH of the formulation is about 5.5 to about 6.8.

17. The stable lyophilized pharmaceutical formulation of claim 16, comprising a bulking agent.

18. The stable lyophilized pharmaceutical formulation of claim 17, wherein the bulking agent is selected from the group consisting of sucrose, trehalose, and mannitol.

19. The stable lyophilized pharmaceutical formulation of claim 17 or 18, wherein the bulking agent is sucrose.

20. The stable lyophilized pharmaceutical formulation of any one of claims 16-19, wherein the buffer is selected from the group consisting of MES, histidine, phosphate, and maleate.

21. The stable lyophilized pharmaceutical formulation of any one of claims 16-20, wherein the buffer is selected from the group consisting of MES and histidine.

22. The stable lyophilized pharmaceutical formulation of any one of claims 16-21, wherein the buffer is MES.

23. The stable lyophilized pharmaceutical formulation of any one of claims 16-22, wherein the formulation is stable at 40°C after storage for at least about 30 days.

24. The stable lyophilized pharmaceutical formulation of any one of claims 16-23, wherein DAR is about 7.4 or greater after storage at 40°C for at least about 30 days.

25. The stable lyophilized pharmaceutical formulation of any one of claims 16-24, wherein % free SN-38 is less than about 3.5 after storage for at least about 30 days at 40°C.

26. The stable lyophilized pharmaceutical formulation of any one of claims 16-25, comprising about 40 mg / mL SG.

27. The stable lyophilized pharmaceutical formulation of any one of claims 16-26, wherein the formulation is stable for at least about 30 days as both a pre-lyophilized formulation at -20°C and as a lyophilized formulation at 40°C.

28. A stable liquid pharmaceutical formulation comprising: about 15 mg / mL - 40 mg / mL SG; a buffer; and a surfactant; wherein pH of the formulation is about 6.0 to about 7.0.

29. The stable liquid pharmaceutical formulation of claim 28, comprising a bulking agent.

30. The stable liquid formulation of claim 28 or 29, wherein the formulation is suitable for large scale production.

31. The stable liquid formulation of any one of claims 28-30, comprising about 10 nM to about 100 nM buffer.

32. The stable liquid formulation of any one of claims 28-31, comprising about 10 nM to about 100 nM bulking agent.

33. The stable liquid formulation of any one of claims 28-32, comprising about 0.01 to about 0.05 % w / v surfactant.

34. The stable liquid formulation of any one of claims 28-33, wherein the buffer is MES.

35. The stable liquid formulation of any one of claims 29-34, wherein the bulking agent is trehalose.

36. The stable liquid formulation of any one of claims 28-35, comprising about 20 mg / mL to about 25 mg / mL SG.

37. The stable liquid formulation of any one of claims 28-36, comprising about 25 mg / mL SG.

38. The stable liquid formulation of any one of claims 28-37, comprising about 10 nM to about 100 nM MES.

39. The stable liquid formulation of any one of claims 28-38, wherein the surfactant is PS80.

40. A pharmaceutical batch comprising sacituzumab govitecan (SG) and about 1.0 pg / mg free drug related impurities or less, wherein the pharmaceutical batch comprises at least about 6.5 kg of SG.

41. The pharmaceutical batch of claim 40, wherein the batch is produced by running a formulation comprising SG through at least two different filtration methods.

42. The pharmaceutical batch of claim 40 or 41, wherein the batch is produced by contacting a solution comprising a drug-linker with a solution comprising an antibody, wherein the concentration of the solution comprising the antibody is greater than about 10 g / L antibody.

43. A pharmaceutical batch consisting essentially of sacituzumab govitecan (SG) and free- drug related impurities resulting from contacting a solution comprising drug-linker with a solution comprising an antibody, wherein the concentration of the solution comprising the antibody is greater than about 10 g / L.

44. A stable lyophilized formulation comprising about 180 mg to about 200 mg sacituzumab govitecan (SG), wherein the formulation comprises less than about 2 pg / mg of free drug-related impurities.

45. A stable lyophilized pharmaceutical formulation comprising about 180 mg to about 200 mg sacituzumab govitecan (SG); and a pharmaceutically acceptable carrier, wherein the formulation is obtained from a plurality of pilot batches wherein median free SN-38 of the plurality of pilot batches is less than or equal to about 2 pg / mg.

46. A method of manufacture of sacituzumab govitecan (SG), comprising the step of subjecting a mixture comprising a crude ADC through at least two different purification filters.

47. The method of claim 46, wherein one of the at least two different purification filters is UF / DF.

48. The method of claim 46 or 47, wherein one of the at least two different purification filters is activated carbon.

49. The method of any one of claims 46-48, comprising a conjugation step prior to the step of subjecting the mixture through at least two different purification filters, wherein the conjugation step comprises a step of contacting an antibody composition comprising greater than 10 g / L hRS7 with drug linker, forming a conjugation mixture having greater than 6 mg / mL hRS7.

50. The method of claim 49, wherein the antibody composition is greater than about 30 g / L hRS7, and wherein the conjugation mixture has greater than about 15 mg / mL hRS7.

51. A method of manufacture of sacituzumab govitecan (SG), comprising a conjugation step, wherein the conjugation step comprises a step of contacting an antibody composition comprising greater than 10 g / L hRS7 with drug linker, forming a conjugation mixture greater than 6 mg / mL hRS7.

52. A pre-lyophilization liquid formulation comprising: a. 10 mg / mL to 40 mg / mL sacituzumab govitecan (SG); and b. a pH of 5.8 to 6.5, wherein the formulation is stable.

53. A stable pre-lyophilization liquid formulation comprising: a. 10 mg / mL to 40 mg / mL of sacituzumab govitecan (SG); and b. a pH of 5.8 to 6.5, wherein the formulation comprises < 1.0 pg / mg free SN-38.

54. A stable pre-lyophilization liquid formulation comprising: a. 10 mg / mL to 40 mg / mL sacituzumab govitecan (SG); and b. a pH of 5.8 to 6.5, wherein the formulation comprises < 3.0% high molecular weight species as measured by SE- UPLC.

55. A stable pre-lyophilization liquid formulation comprising:a. 10 mg / mL to 40 mg / mL SG; and b. a pH of 5.8 to 6.5; wherein the formulation has a DAR of 7.6 as measured by native LC / MS.

56. A stable pre-lyophilization liquid formulation comprising: a. 10 mg / mL to 40 mg / mL sacituzumab govitecan (SG); b. a buffer; c. a bulking agent; and d. a surfactant; wherein pH of the formulation is 5.8 to 6.0.

57. A stable pre-lyophilization liquid formulation of sacituzumab govitecan (SG), wherein free SN-38 is < 1.0 pg / mg for 12 months at -< 30°C.

58. A stable pre-lyophilization liquid formulation comprising: a. 15 mg / mL to 40 mg / mL SG; b. a buffer; c. a bulking agent; and d. a surfactant, wherein pH of the formulation is 5.8 to 6.5.

59. A pre-lyophilization liquid formulation of sufficient stability to limit free SN-38 to 1.0 < pg / mg for 1 month at 2-8°C, wherein the formulation has a pH of 5.8 to 6.0.

60. A pre-lyophilization liquid formulation comprising: a. < 1.0 pg / mg free SN-38; b. < 1.0 pg / mg free drug-related impurities; c. < 3.0 % HMW species; and d. a pH of 5.8 to 6.0, wherein the formulation is stable.

61. The formulation of any one of claims 52-60, wherein when the formulation is lyophilized and reconstituted, the formulation has < 1.0 pg / mg free SN-38.

62. A vial comprising less than 20 mL of the formulation of any one of claims 52-61, wherein the vial comprises 180 mg to 200 mg SG after the formulation is lyophilized.

63. A vial comprising 8 mL of the formulation of any one of claims 52-61, wherein the vial comprises 180 mg to 200 mg SG after lyophilization.

64. The vial of claim 61 or 62, wherein the vial is a 30R vial.

65. A batch or lot of 1000 vials, each vial comprising a lyophilized formulation of 180 mg to 200 mg SG and a pharmaceutically acceptable carrier, wherein median or average free SN-38 of the batch or lot is < 1.0 pg / mg.

66. The batch or lot of claim 65, wherein the vials are obtainable from a 10 kg pharmaceutical batch.

67. A stable lyophilized formulation comprising: a. 180 mg to 200 mg SG; and b. a pharmaceutically acceptable carrier, wherein the stable lyophilized formulation is obtained from or obtainable from a pharmaceutical batch of 10 kg SG, wherein the pharmaceutical batch comprises < 1.0 pg / mg free drug-related impurities at drug substance release.

68. A reconstituted pharmaceutical formulation suitable for intravenous administration comprising 10 mg / mL sacituzumab govitecan (SG), wherein pH of the formulation is 5.8 to 6.0.

69. A lyophilized formulation of SG, wherein the formulation is made by lyophilizing an aqueous composition comprising: a. 10 mg / mL to 70 mg / mL of sacituzumab govitecan (SG); b. a buffer for maintaining pH at 5.8 to 6.0; and c. a surfactant, wherein the formulation is stable.

70. A stable lyophilized pharmaceutical composition comprising 180 mg to 200 mg sacituzumab govitecan (SG), and a pharmaceutically acceptable carrier, wherein the composition comprises < 0.5 pg / mL free SN-38 at 6 months at 5°C.

71. A stable lyophilized formulation comprising 180 mg to 200 mg sacituzumab govitecan (SG), wherein the lyophilized formulation is obtainable from a pharmaceutical batch comprising 10 kg SG, wherein the pharmaceutical batch comprises < 1.0 pg / mg free SN-38 at drug substance release.

72. A lyophilized formulation of 180 mg to 200 mg sacituzumab govitecan (SG), the lyophilized formulation obtainable or obtained by lyophilizing an aqueous formulation said SG and pharmaceutically acceptable excipients, wherein the aqueous formulation comprises a buffer that maintains pH during lyophilization from 5.8 to 6.3.

73. A lyophilized formulation of 180 mg to 200 mg sacituzumab govitecan (SG), the lyophilized formulation obtainable or obtained by lyophilizing an aqueous formulation of said SG and pharmaceutically acceptable excipients, wherein the aqueous formulation comprises 15 mg / mL to 40 mg / mL SG and wherein the aqueous formulation comprises a buffer that maintains shifts in pH during lyophilization from 5.8 to 6.5.

74. A stable lyophilized formulation comprising 180 mg to 200 mg sacituzumab govitecan (SG) obtained from a 10 kg pharmaceutical batch of SG.

75. A lyophilized pharmaceutical formulation of 180 mg to 200 mg sacituzumab govitecan (SG), wherein the lyophilized pharmaceutical formulation is stable for 12 months at 2-8°C at a residual moisture content of < 3.7%.

76. A pharmaceutical batch comprising 10 kg sacituzumab govitecan (SG), wherein the pharmaceutical batch yield is at least about 97% and wherein the batch comprises < 1.0 pg / mg free drug-related impurities.

77. The pharmaceutical batch of claim 76, wherein the batch is produced by running a composition comprising SG through at least two different filtration methods.

78. The pharmaceutical batch of claim 76 or 77, wherein the pharmaceutical batch is produced by contacting a solution comprising CL2A-SN-38 with a solution comprising hRS7, wherein the concentration of the solution comprising hRS7 is greater than 10 g / L hRS7.

79. A stable antibody formulation comprising: a. 50 g / L hRS7; and b. a buffer, wherein the formulation stable at 2-8°C for 36 months.

80. A stable antibody formulation comprising: a. 50 g / L hRS7; and b. a buffer, wherein the formulation stable at < -30°C for 36 months.

81. A method of manufacturing a composition comprising sacituzumab govitecan (SG), wherein the method comprises: a. pooling mAb (i.e., hRS7) at a concentration greater than about 10 g / L in a buffer; b. reducing the pooled mAb for at least about 4 hours to produce reduced mAb; c. conjugating the reduced mAb with a conjugation mixture to produce a crude antibody-drug conjugate (ADC), wherein the conjugation mixture comprises greater than about 6 mg / mL reduced mAb and about 10 molar equivalents of drug-linker; and d. purifying SG from the crude ADC using at least two different purification methods.

82. The method of claim 81, wherein step (a) comprises pooling mAb at a concentration of 50 g / L or 60 g / L.

83. The method of claim 81 or 82, wherein step (a) comprises pooling mAb at a concentration greater than 10 g / L in a buffer comprising sodium acetate.

84. The method of any one of claims 81-83, wherein the conjugation mixture comprises 30 g / L reduced mAb.

85. The method of any one of claims 81-84, wherein the conjugation produces a crude ADC comprising SG at a concentration of 15-45 mg / mL SG.

86. The method of any one of claims 81-85, wherein SG is purified from the crude ADC using activated carbon or carbon depth filtration.

87. The method of any one of claims 81-85, wherein SG is purified from the crude ADC using ultrafiltration / diafiltration.

88. A composition comprising sacituzumab govitecan (SG) obtainable by the method of any one of claims 81-87.

89. The composition of claim 88, wherein the composition comprises greater than 10 mg / mL SG, such as about 25 mg / mL SG.

90. A pre-lyophilization liquid pharmaceutical formulation comprising sacituzumab govitecan (SG), wherein the formulation comprises greater than 10 mg / mL SG, a surfactant, and a buffer, and wherein:(i) the DAR of the formulation is no lower than 7.35 when the formulation is stored for at least about 30 days at a temperature of -20°C; and / or(ii) the formulation comprises less than about 10 pg / mg free SN-38 when the formulation is stored for at least about 30 days at a temperature of -20°C.

91. The pre-lyophilization liquid pharmaceutical formulation of claim 90, wherein the formulation comprises about 20 mg / mL SG to about 40 mg / mL SG, such as about 25 mg / mL SG.

92. The pre-lyophilization liquid pharmaceutical formulation of claim 90 or 91, wherein the surfactant is polysorbate 80 and the formulation comprises about 0.02 %w / v to about 0.04 %w / v polysorbate 80, such as about 0.025%w / v polysorbate 80.

93. The pre-lyophilization liquid pharmaceutical formulation of any one of claims 90-92, wherein the buffer is MES and the formulation comprises about 41 mM to about 82 mM MES, such as about 51 mM MES. ,94. The pre-lyophilization liquid pharmaceutical formulation of any one of claims 90-93, wherein the formulation also comprises trehalose, and the formulation comprises about 50 mM to about 100 mM trehalose, such as about 62.5 mM trehalose.

95. A kit comprising less than 20 ml, such as 8 ml of the pre-lyophilization liquid pharmaceutical formulation of any one of claims 90-94 in a 20R vial.

96. A pre-lyophilization liquid formulation drug substance comprising: an ADC of the structure of Formula I and a means for stabilizing said ADC in said liquid formulation drug substance.

97. A reconstituted liquid formulation drug product comprising an ADC of Formula I:, and means for stabilizing said liquid formulation drug product.

98. A drug product comprising: a lyophilized ADC comprising an ADC of Formula I:, and reconstituted ADC in a liquid formulation drug product, and means for achieving in said liquid formulation drug product a desired concentration of SN-38 and a desired stability of said ADC.

99. An ADC comprising: a. monoclonal humanized antibody means that bind to Trop-2 Trophoblast cellsurface antigen-2; b. cytotoxic topoisomerase inhibitor means, andc. means for linking said cytotoxic means to said antibody means, wherein said ADC is capable of a concentration of 40 mg / ml in a liquid formulation, a pH of 5.8, and a lyophilization cycle less than 4 days.

100. A reconstituted liquid formulation drug product comprising: a. monoclonal humanized antibody means that bind to Trop-2 Trophoblast cellsurface antigen-2; b. cytotoxic topoisomerase inhibitor means; c. means for linking said cytotoxic means to said antibody means; d. surfactant means; e. buffer means; and f. bulking agent means, wherein said liquid formulation drug product is capable of: i. a concentration of 40 mg / ml in a liquid formulation; ii. a pH of 5.8; iii. a lyophilization cycle less than 4 days; iv. stability at -20°C after storage for at least about 30 days; v. a DAR of 7.4 or greater after storage at -20°C for at least about 30 days; vi. % HMW species stability for at least about 30 days at -20°C; vii. a DAR of 7.3 or greater after storage at 5°C for about 60 hours; and / or viii. a stable DAR for at least about 30 days as both a liquid formulation at - 20°C and as a lyophilized formulation at 40°C.

101. A drug substance comprising an ADC for Formula I:I l lproduced by purifying the ADC through a carbon filter.

102. A drug product comprising an ADC of Formula I:produced by purifying the ADC through a carbon filter.

103. A method of treating a human patient in need of treatment for: a. unresectable locally advanced or metastatic triple-negative breast cancer, wherein said patient has previously received two or more prior systemic therapies, at least one of them for metastatic disease; and / orb. unresectable locally advanced or metastatic HR-positive, HER2- (IHC 0, IHC 1+, or IHC 2+ / ISH-) breast cancer, wherein said patient has received endocrine-based therapy and at least two additional systemic therapies in the metastatic setting, wherein said method comprises administering to said patient an amount effective for said treatment a reconstituted liquid formulation drug product comprising an ADC of Formula I:means for stabilizing said liquid formulation drug product.

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