Crystalline antibody formulations

Patent metadata

Field	Value
Publication number	US-2017198059-A1
Application number	US-201515326341-A
Country	US
Kind code	A1
Filing date	Jul 13, 2015
Priority date	Jul 14, 2014
Publication date	Jul 13, 2017
Grant date	—

Abstract

Described herein are anti-PCSK9 antibody crystals, methods of making such antibody crystals and formulations comprising the antibody crystals.

First claim

1 . A crystal of an anti-PCSK9 IgG antibody comprising a light chain complementarity determining region (CDR) of the CDRL1 sequence in SEQ ID NO:9, a CDRL2 of the CDRL2 sequence in SEQ ID NO:9, and a CDRL3 of the CDRL3 sequence in SEQ ID NO:9, and a heavy chain complementarity determining region (CDR) of the CDRH1 sequence in SEQ ID NO:5, a CDRH2 of the CDRH2 sequence in SEQ ID NO:5, and a CDRH3 of the CDRH3 sequence in SEQ ID NO:5.

2 . The crystal of claim 1 , wherein the anti-PCSK9 IgG antibody comprises a light chain variable region that is at least 90% identical to that of SEQ ID NO:9 or SEQ ID NO:11 and a heavy chain variable region that is at least 90% identical to that of SEQ ID NO:5 or SEQ ID NO:7.

3 . The crystal of claim 2 , wherein the anti-PCSK9 IgG antibody comprises a light chain variable region having the amino acid sequence set forth in SEQ ID NO:9 or SEQ ID NO:11 and a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 or SEQ ID NO:7.

4 . The crystal of claim 1 , wherein the crystal has a length of about 5 μM to about 50 μM.

5 . The crystal of claim 1 wherein the crystal has a rod or needle shape.

6 . The crystal of any of claim 1 , wherein the crystal comprises a salt selected from the group consisting of sodium di-hydrogen phosphate, di-potassium hydrogen phosphate, sodium chloride, ammonium sulfate, potassium sodium tartrate tetrahydrate, sodium citrate dihydrate, sodium acetate trihydrate, di-ammonium hydrogen phosphate, potassium sodium tartrate, calcium acetate, cacodylate, CHES, CAPS, Tris, lithium sulfate, sodium phosphate, potassium phosphate, sodium sulfate.

7 . A method of making a crystal of an anti-PCSK-9 antibody comprising combining a solution of the antibody with a crystallization reagent comprising a salt selected from the group consisting of sodium di-hydrogen phosphate, di-potassium hydrogen phosphate, sodium chloride, ammonium sulfate, potassium sodium tartrate tetrahydrate, sodium citrate dihydrate, sodium acetate trihydrate, di-ammonium hydrogen phosphate, potassium sodium tartrate, calcium acetate, cacodylate, CHES, CAPS, Tris, lithium sulfate, sodium phosphate, potassium phosphate, and sodium sulfate in a crystallization buffer, wherein the anti-PCSK-9 antibody is selected from the group consisting of an antibody comprising:
(a) a light chain variable region having the amino acid sequence set forth in SEQ ID NO:9 or SEQ ID NO:11 and a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 or SEQ ID NO:7; (b) a light chain variable region that is at least 90% identical to that of SEQ ID NO:9 or SEQ ID NO:11 and a heavy chain variable region that is at least 90% identical to that of SEQ ID NO:5 or SEQ ID NO:7; (c) a light chain complementarity determining region (CDR) of the CDRL1 sequence in SEQ ID NO:9, a CDRL2 of the CDRL2 sequence in SEQ ID NO:9, and a CDRL3 of the CDRL3 sequence in SEQ ID NO:9, and a heavy chain complementarity determining region (CDR) of the CDRH1 sequence in SEQ ID NO:5, a CDRH2 of the CDRH2 sequence in SEQ ID NO:5, and a CDRH3 of the CDRH3 sequence in SEQ ID NO:5; and (d) a light chain complementarity determining region of the CDRL1 sequence of SEQ ID NO:24, the CDRL2 sequence of SEQ ID NO:25, the CDRL3 sequence of SEQ ID NO:26, and a heavy chain complementarity determining region (CDR) of the CDRH1 sequence of SEQ ID NO:20 or SEQ ID NO:21, the CDRH2 sequence of SEQ ID NO:22, and the CDRH3 sequence of SEQ ID NO:23.

8 . The method of claim 7 , wherein the concentration of salt in the crystallization buffer is from about 0.1M to about 10M.

9 . The method of claim 7 , further comprising removing at least a portion of the crystallization buffer after crystals have formed.

10 . The method of claim 9 , wherein the portion of crystallization buffer is removed by centrifugation.

11 . The method of claim 10 , wherein the crystals are placed in a solution containing an organic additive.

12 . The method of claim 11 , further comprising the addition of an excipient to the solution.

13 . The method of claim 7 , further comprising drying crystals that have formed.

14 . The method of claim 13 , wherein the crystals are dried by exposure to air, or by exposure to a vacuum, or by exposure to nitrogen gas.

15 . An antibody crystal produced by the method of claim 7 .

16 . A crystalline formulation of the antibody of claim 1 .

17 . A method of lowering serum LDL cholesterol or treating a disorder associated with increased levels of serum LDL cholesterol in a mammalian subject in need thereof comprising administering the crystal of claim 1 or the crystalline formulation of claim 16 in an amount effective to lower serum LDL cholesterol levels in the subject as compared to a predose serum LDL cholesterol level.

18 . The method of claim 7 , wherein the crystallization buffer further comprises polyethylene glycol (PEG).

19 . The method of claim 18 , wherein the PEG has a molecular weight of about 400 kDa to about 20,000 kDa.

20 . The method of claim 19 , wherein the PEG is present at a concentration of about 0.1% to about 50%.

21 . The crystalline formulation of claim 16 , wherein the crystal comprises a salt selected from the group consisting of sodium dihydrogen phosphate, di-potassium hydrogen phosphate, sodium chloride, ammonium sulfate, potassium sodium tartrate tetrahydrate, tacsimate, sodium citrate dihydrate, sodium acetate trihydrate, di-ammonium tartrate, sodium malonate, acetate, calcium acetate, cacodylate, CHES, lithium sulfate, magnesium chloride, zinc acetate, cesium chloride, ammonium phosphate, sodium phosphate, potassium phosphate, sodium fluoride, potassium iodide, sodium idodide, ammonium iodide, sodium thiocyanate, potassium thiocyanate, sodium formate, potassium formate, and ammonium formate.

22 . The crystalline formulation of claim 16 , wherein the crystals have a length of about 5 μm to about 50 μm and a morphology selected from the group consisting of rod shapes and needle shapes, or a mixture thereof.

23 . The crystalline formulation of claim 16 , comprising a crystal of any one of claims 1 - 6 .

24 . The crystalline formulation of claim 16 , comprising at least 20% PEG3350 precipitant.

25 . The crystalline formulation of claim 16 , wherein the osmolality of the formulation ranges from about 180 to about 420 mOsm/kg.

26 . A container comprising at least 100 to about 450 mg or more of the crystals of claim 6 for reconstitution in a volume of 0.5-2 mL.

27 . A container comprising the crystalline formulation of claim 16 at a concentration of at least 100 mg/ml.

Assignees

• Amgen Inc

Inventors

• Khalaf Nazer
• Mandapati Saraswathi
• Patel Reena J

Classifications

• A61K39/39591

  Stabilisation, fragmentation · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title

• C07K16/40 Primary

  against enzymes · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title