Lyophilized anti-tissue factor antibody-drug conjugates
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. 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 lyophilized formulation is free of surfactant,
(b) 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
(c) the drug portion of the ADC comprises vcMMAE.
2. The lyophilized formulation of claim 1 , 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.
3. The lyophilized formulation of claim 1 , 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.
4. The lyophilized formulation of claim 2 , wherein the aqueous formulation comprises the buffer at a concentration of about 20 to about 50 mM.
5. The lyophilized formulation of claim 2 , wherein the non-reducing sugar is selected from sucrose, trehalose and a combination thereof.
6. The lyophilized formulation of claim 2 , wherein the aqueous formulation comprises the non-reducing sugar at a concentration of about 10 to about 250 mM.
7. The lyophilized formulation of claim 2 , wherein the bulking agent is selected from mannitol and glycine.
8. The lyophilized formulation of claim 2 , wherein the aqueous formulation comprises the bulking agent at a concentration of about 50 mM to about 300 mM.
9. The lyophilized formulation of claim 1 , wherein the aqueous formulation comprises from about 5 g/L to about 30 g/L anti-TF ADC.
10. The lyophilized formulation of claim 1 , wherein the pH of the aqueous formulation is in a range from about 5.5 to 6.5.
11. The lyophilized formulation of claim 1 , comprising mannitol and sucrose, wherein the weight by weight ratio of mannitol to sucrose is between about 1:1 and about 30:1.
12. The lyophilized formulation of claim 1 , 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.
13. The lyophilized formulation of claim 1 , 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.
14. The lyophilized formulation of claim 1 , wherein the aqueous formulation comprises from about 9 to about 11 g/L anti-TF ADC.
15. The lyophilized formulation of claim 1 , 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.
16. The lyophilized formulation of claim 1 , wherein the average absolute number of drug moieties per antibody molecule is 1, 2, 3, 4, 5, 6, 7, or 8.
17. The lyophilized formulation of claim 1 , wherein the anti-TF antibody is a full-length antibody.
18. The lyophilized formulation of claim 1 , 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.
19. The lyophilized formulation of claim 1 , wherein the anti-TF ADC is stable at 2-8° C. for pharmaceutical use for at least 6 months.
20. The lyophilized formulation of claim 19 , wherein the formulation is stable when it has less than 3.0% aggregates when stored at 5° C. for at least 6 months.
21. The lyophilized formulation of claim 19 wherein the stability is determined by SEC analysis according to Example 10.
22. The lyophilized formulation of claim 1 , which lyophilized formulation contains less than 3.0 wt. % moisture.
23. The lyophilized formulation of claim 1 , wherein the formulation is free of any inorganic salts.
24. 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 1 ;
b. about 28 to 34 mM histidine;
c. about 84 to about 146 mM sucrose;
d. about 158 to about 274 mannitol.
25. A pharmaceutically acceptable liquid formulation obtained by reconstituting the lyophilized formulation of claim 1 in a sterile aqueous diluent.
26. The liquid formulation of claim 25 , 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.
27. A method for preparing a lyophilized formulation of claim 1 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.
28. A method of preparing an injectable solution of an anti-TF ADC, comprising the step of reconstituting the lyophilized formulation of claim 1 in a sterile aqueous diluent.