Precision Medicine by targeting rare human PCSK9 variants for cholesterol treatment

• US 9,040,052 B1
• Filed: 09/18/2014
• Issued: 05/26/2015
• Est. Priority Date: 12/17/2013
• Status: Active Grant

• Current Assignee: Kymab Limited
• Original Assignee: Kymab Limited
• Inventors: Clube, Jasper Rupert
• Primary Examiner(s): Wen, Sharon
• Application Number:
  US14/490,175
• Publication Number:
  US 20150166672A1
• Time in Patent Office: 250 Days
• Field of Search: None
• US Class Current: 424/146.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/62 comprising only variable region components
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material

Abstract

The invention relates to human targets of interest (TOI), anti-TOI ligands, kits compositions and method.

Claims

1. A method of reducing cholesterol level or maintaining previously reduced cholesterol level in a human in need thereof, the method comprising administering to said human an antibody or antibody fragment that specifically binds a proprotein convertase subtilisin/kexin type 9 (PCSK9) that comprises a C-terminal domain comprising a mutation selected from the group consisting of I474V and E670G in SEQ ID NO:

1,

wherein the antibody or antibody fragment comprises a VH domain derived from the recombination of a human VH segment, a human D gene segment and a human JH segment, the human VH segment encoding the framework 1 of SEQ ID NO;

40, and wherein said human comprises;

(i) a VH gene segment encoding the framework 1 of SEQ ID NO;

40, or the human expresses antibodies comprising VH domains derived from the recombination of a human VH segment, a human D gene segment and a human JH segment, the human VH segment encoding the framework 1 of SEQ ID NO;

40; and

(ii) a nucleotide sequence encoding said PCSK9 that comprises a C-terminal domain comprising said selected mutation in SEQ ID NO;

1.

2. The method of claim 1, wherein each said VH gene segment is VH3-23*04 of SEQ ID NO:

39.

3. The method of claim 1, wherein the antibody comprises a VH domain, wherein the VH domain comprises the framework 1 sequence of SEQ ID NO:

40.

4. The method of claim 1, comprising, before said administering, selecting a human comprising (i) said nucleotide sequence encoding said PCSK9 that comprises a C-terminal domain comprising a mutation I474V or E670G in SEQ ID NO:

1, and/or (ii) a PCSK9 variant protein encoded by the nucleotide sequence of SEQ ID NO;

30.

5. The method of claim 1, wherein the human has been determined to comprise (i) the nucleotide sequence encoding the PCSK9 C-terminal domain comprising a mutation I474V or E670G in SEQ ID NO:

1, and/or (ii) a PCSK9 variant protein encoded by the nucleotide sequence of SEQ ID NO;

30.

6. The method of claim 1, comprising the step of determining that the human comprises (i) the nucleotide sequence encoding the PCSK9 C-terminal domain comprising a mutation I474V or E670G in SEQ ID NO:

1, and/or (ii) a PCSK9 variant protein encoded by the nucleotide sequence of SEQ ID NO;

30, wherein the determining step is performed before administration of the antibody to the human.

7. The method of claim 6, wherein the step of determining comprises assaying a biological sample from the human for a nucleotide sequence encoding the PCSK9 that comprises the C-terminal domain comprising the mutation I474V or E670G in SEQ ID NO:

1.

8. The method of claim 7, wherein the assaying comprises contacting the biological sample with

a. at least one oligonucleotide probe comprising a sequence of at least 10 contiguous nucleotides that can specifically hybridize to and identify in the biological sample a nucleotide sequence encoding the PCSK9 that comprises the C-terminal domain comprising the mutation I474V or E670G in SEQ ID NO:

1, or that specifically hybridizes to an antisense of said sequence, wherein said nucleic acid hybridizes to at least one nucleotide present in said selected sequence which is not present in SEQ ID NO;

28 or hybridizes to an antisense sequence thereby forming a complex when at least one nucleotide sequence encoding the PCSK9 that comprises the C-terminal domain comprising the mutation I474V or E670G in SEQ ID NO;

1 is present; and

/or

b. at least one oligonucleotide probe comprising a sequence of at least 10 contiguous nucleotides of a nucleotide sequence encoding the PCSK9 that comprises the C-terminal domain comprising the mutation I474V or E670G in SEQ ID NO;

1 comprising an antisense sequence of said contiguous nucleotides, wherein said sequence of contiguous nucleotides comprises at least one nucleotide present in said selected sequence which is not present in SEQ ID NO;

28 thereby forming a complex when the nucleotide sequence encoding the PCSK9 that comprises a C-terminal domain comprising the mutation I474V or E670G in SEQ ID NO;

1 is present; and

detecting the presence or absence of the complex, wherein detecting the presence of the complex determines that the human comprises the PCSK9 that comprises the C-terminal domain comprising the mutation I474V or E670G in SEQ ID NO;

1.

9. The method of claim 8, wherein the assaying comprises nucleic acid amplification and optionally one or more methods selected from sequencing, next generation sequencing, nucleic acid hybridization, and allele-specific amplification and/or wherein the assaying is performed in a multiplex format.

10. The method of claim 1, wherein said human is or has been further determined to be substantially resistant to statin treatment.

11. The method of claim 1, wherein the human is receiving or has received statin treatment or has reduced responsiveness to statin treatment.

12. The method of claim 5, wherein said human is or has been further determined to be substantially resistant to statin treatment.

13. The method of claim 5, wherein the human is receiving or has received statin treatment or has reduced responsiveness to statin treatment.

14. The method of claim 1, wherein said antibody or antibody fragment is administered separately or simultaneously with a statin treatment.

15. The method of claim 8, wherein said antibody or antibody fragment is administered separately or simultaneously with a statin treatment.

16. The method of claim 1, wherein said human is indicated as heterozygous for a nucleotide sequence encoding the PCSK9 C-terminal domain comprising a mutation E670G, optionally, wherein said human is further indicated as comprising the nucleotide sequence of SEQ ID NO:

28, or said human is indicated as homozygous for a nucleotide sequence encoding the PCSK9 C-terminal domain comprising a mutation I474V or E670G in SEQ ID NO;

1.

17. The method of claim 1, wherein said human has been diagnosed with at least one condition selected from a lipid disorder, hyperlipoproteinemia, hyperlipidemia, dyslipidemia, hypercholesterolemia, a heart attack, a stroke, coronary heart disease, atherosclerosis, peripheral vascular disease, claudication, and high blood pressure.

18. The method of claim 1, wherein said antibody or antibody fragment treats or reduces the risk of in said human at least one condition selected from a lipid disorder, hyperlipoproteinemia, hyperlipidemia, dyslipidemia, hypercholesterolemia, a heart attack, a stroke, coronary heart disease, atherosclerosis, peripheral vascular disease, claudication, and high blood pressure.

19. The method of claim 15, wherein said human has been diagnosed with at least one condition selected from a lipid disorder, hyperlipoproteinemia, hyperlipidemia, dyslipidemia, hypercholesterolemia, a heart attack, a stroke, coronary heart disease, atherosclerosis, peripheral vascular disease, claudication, and high blood pressure.

20. The method of claim 15, wherein said antibody or antibody fragment treats or reduces the risk of in said human at least one condition selected from a lipid disorder, hyperlipoproteinemia, hyperlipidemia, dyslipidemia, hypercholesterolemia, a heart attack, a stroke, coronary heart disease, atherosclerosis, peripheral vascular disease, claudication, and high blood pressure.

21. The method of claim 1, wherein the nucleotide sequence encoding the PCSK9 C-terminal domain comprising a mutation I474V or E670G is SEQ ID NO:

30.

22. The method of claim 1, wherein the nucleotide sequence is SEQ ID NO:

30.

23. The method of claim 1, wherein the step of administering is performed by intravenous or subcutaneous administration.

24. The method of claim 15, wherein the step of administering is performed by intravenous or subcutaneous administration.

25. The method of claim 1, wherein the antibody or antibody fragment is comprised in an injectable preparation.

26. The method of claim 1, wherein the antibody is a human antibody.