Antibody-drug conjugate lyophilised formulation

Abstract

Disclosed herein are surfactant free lyophilized formulations of antibody-drug conjugates (ADCs), such as anti-tissue factor ADCs, and reconstituted formulations, processes and uses thereof. The formulations are particularly suitable for an anti-TF ADC based on an auristatin derivative or other similarly hydrophobic drug. Typically, the excipients of the formulations comprise or consist of histidine, sucrose, trehalose, mannitol and glycine.

1 - 45 . (canceled) 
     
     
         46 . A lyophilized formulation of an anti-tissue factor (TF) antibody-drug conjugate (ADC), the lyophilized formulation obtainable or obtained by lyophilizing an aqueous formulation comprising said anti-TF ADC and pharmaceutically acceptable excipients, wherein:
 (a) the anti-TF antibody portion of the ADC comprises a variable heavy (VH) region comprising a CDR1 region having the amino acid sequence set forth in SEQ ID NO: 6, a CDR2 region having the amino acid sequence set forth in SEQ ID NO: 7, and a CDR3 region having the amino acid sequence set forth in SEQ ID NO: 8, and a variable light (VL) region comprising a CDR1 region having the amino acid sequence set forth in SEQ ID NO:46, a CDR2 region having the amino acid sequence set forth in SEQ ID NO: 47, and a CDR3 region having the amino acid sequence set forth in SEQ ID NO: 48, and   (b) the drug portion of the ADC comprises vcMMAE.   
     
     
         47 . The lyophilized formulation of  claim 46 , wherein the pharmaceutically acceptable excipients comprise:
 a) a buffer which limits pH shifts during the lyophilizing step so that pH is kept between about 5 and about 7,   b) at least one non-reducing sugar which forms an amorphous phase with the anti-TF ADC in solid state; and   c) at least one bulking agent.   
     
     
         48 . The lyophilized formulation of  claim 47 , wherein the aqueous formulation comprises a buffer selected from the group consisting of histidine, citrate, phosphate, carbonic acid, succinate, glycolate and a combination of any thereof. 
     
     
         49 . The lyophilized formulation of  claim 47 , wherein the aqueous formulation comprises the buffer at a concentration of about 20 to about 50 mM. 
     
     
         50 . The lyophilized formulation of  claim 47 , wherein the non-reducing sugar is selected from sucrose, trehalose and a combination thereof. 
     
     
         51 . The lyophilized formulation of  claim 47 , wherein the aqueous formulation comprises the non-reducing sugar at a concentration of about 10 to about 250 mM. 
     
     
         52 . The lyophilized formulation of  claim 47 , wherein the bulking agent is selected from mannitol and glycine. 
     
     
         53 . The lyophilized formulation of  claim 47 , wherein the aqueous formulation comprises the bulking agent at a concentration of about 50 mM to about 300 mM. 
     
     
         54 . The lyophilized formulation of  claim 46 , wherein the aqueous formulation comprises from about 5 g/L to about 30 g/L anti-TF ADC. 
     
     
         55 . The lyophilized formulation of  claim 46 , wherein the pH of the aqueous formulation is in a range from about 5.5 to 6.5. 
     
     
         56 . The lyophilized formulation of  claim 46 , comprising mannitol and sucrose, wherein the weight by weight ratio of mannitol to sucrose is between about 1:1 and about 30:1. 
     
     
         57 . The lyophilized formulation of  claim 46 , comprising mannitol and sucrose, wherein the weight by weight ratio of mannitol to anti-TF ADC is about 3:1 and the weight by weight ratio of mannitol to sucrose is about 1:1. 
     
     
         58 . The lyophilized formulation of  claim 46 , which is obtainable or obtained by lyophilizing an aqueous formulation comprising about 5 g/L to about 30 g/L anti-TF ADC and pharmaceutically acceptable excipients comprising:
 a. about 20 to about 50 mM histidine or citrate buffer having a pH of about 5 to about 7;   b. about 10 to about 250 mM sucrose or trehalose; and   c. about 50 mM to about 300 mM mannitol or glycine.   
     
     
         59 . The lyophilized formulation of  claim 46 , wherein the aqueous formulation comprises from about 9 to about 11 g/L anti-TF ADC. 
     
     
         60 . The lyophilized formulation of  claim 46 , wherein the antibody comprises a VH region comprising an amino acid sequence of SEQ ID NO: 5 and a VL region comprising an amino acid sequence of SEQ ID NO: 45. 
     
     
         61 . The lyophilized formulation of  claim 46 , wherein the average absolute number of drug moieties per antibody molecule is 1, 2, 3, 4, 5, 6, 7, or 8. 
     
     
         62 . The lyophilized formulation of  claim 46 , wherein the anti-TF antibody is a full-length antibody. 
     
     
         63 . The lyophilized formulation of  claim 46 , which is obtainable or obtained by lyophilizing an aqueous formulation comprising about 9 g/L to about 11 g/L anti-TF ADC and pharmaceutically acceptable excipients comprising: about 30 mM histidine buffer having a pH of about 5 to about 7; about 88 mM sucrose; and about 165 mM mannitol; wherein the antibody comprises a VH region comprising an amino acid sequence of SEQ ID NO: 5 and a VL region comprising an amino acid sequence of SEQ ID NO: 45. 
     
     
         64 . The lyophilized formulation of  claim 46 , wherein the anti-TF ADC is stable at 2-8° C. for pharmaceutical use for at least 6 months. 
     
     
         65 . The lyophilized formulation of  claim 46 , which lyophilized formulation contains less than 3.0 wt % moisture. 
     
     
         66 . The lyophilized formulation of  claim 46 , wherein the formulation is free of any inorganic salts. 
     
     
         67 . An aqueous solution suitable for preparing a lyophilized formulation of an anti-TF ADC, comprising:
 a. from about 7 to about 20 g/L anti-TF ADC of  claim 46 ;   b. about 28 to 34 mM histidine;   c. about 84 to about 146 mM sucrose;   d. about 158 to about 274 mannitol.   
     
     
         68 . A pharmaceutically acceptable liquid formulation obtained by reconstituting the lyophilized formulation of  claim 46  in a sterile aqueous diluent. 
     
     
         69 . The liquid formulation of  claim 68 , comprising about 5 g/L to about 30 g/L anti-TF ADC, about 20 to about 50 mM histidine having a pH of about 5 to about 7; about 10 to about 250 mM sucrose or trehalose; and about 50 mM to about 300 mM mannitol or glycine. 
     
     
         70 . A method for preparing a lyophilized formulation of  claim 46  comprising the steps of:
 a. cooling the aqueous solution at a rate of from 0.5° C./min to 1° C./min to a temperature of −40° C. or less; 
 b. holding isothermally for at least 120 min; 
 c. warming to between −20° C. and −15° C. at a rate of from 0.5° C./min to 3° C./min; 
 d. holding isothermally for at least 180 min; 
 e. applying vacuum using a pressure between 50 mTorr and 200 mTorr at a temperature between −30° C. and −10° C.; 
 f. increasing the temperature to between 35° C. and 50° C. at a rate of from 0.5° C./min and 1° C./min and 
 g. holding isothermally for at least 10 hours. 
 
     
     
         71 . A method of preparing an injectable solution of an anti-TF ADC, comprising the step of reconstituting the lyophilized formulation of  claim 46  in a sterile aqueous diluent. 
     
     
         72 . A lyophilized formulation of an anti-tissue factor (TF) antibody-drug conjugate (ADC), the lyophilized formulation obtainable or obtained by lyophilizing an aqueous formulation comprising said anti-TF ADC and pharmaceutically acceptable excipients, wherein:
 (a) the anti-TF antibody portion of the ADC comprises a variable heavy (VH) region comprising a CDR1 region having the amino acid sequence set forth in SEQ ID NO: 6, a CDR2 region having the amino acid sequence set forth in SEQ ID NO: 7, and a CDR3 region having the amino acid sequence set forth in SEQ ID NO: 8, and a variable light (VL) region comprising a CDR1 region having the amino acid sequence set forth in SEQ ID NO:46, a CDR2 region having the amino acid sequence set forth in SEQ ID NO: 47, and a CDR3 region having the amino acid sequence set forth in SEQ ID NO: 48, wherein the anti-TF antibody is an IgG antibody; and   (b) the drug portion of the ADC comprises vcMMAE.   
     
     
         73 . The lyophilized formulation of  claim 72 , wherein the pharmaceutically acceptable excipients comprise:
 a) a buffer which limits pH shifts during the lyophilizing step so that pH is kept between about 5 and about 7,   b) at least one non-reducing sugar which forms an amorphous phase with the anti-TF ADC in solid state; and   c) at least one bulking agent.   
     
     
         74 . The lyophilized formulation of  claim 72 , wherein the lyophilized formulation is free of surfactant.