Targeting rare human PCSK9 variants for cholesterol treatment

• US 10,618,971 B2
• Filed: 09/29/2014
• Issued: 04/14/2020
• 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 X
• Application Number:
  US14/500,397
• Publication Number:
  US 20150166673A1
• Time in Patent Office: 2,024 Days
• Field of Search: None
• 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 comprising

administering to said human an alirocumab antibody or antibody fragment thereof that specifically binds

(i) a proprotein convertase subtilisin/kexin type 9 (PCSK9) amino acid sequence encoded by a PCSK9 nucleotide sequence selected from the group consisting of SEQ ID NOs:

29-37;

or

(ii) an amino acid sequence comprising at least the catalytic domain or C-terminal domain of a PCSK9 protein encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOs;

29-37;

wherein the antibody or antibody fragment comprises;

(iii) a VH domain derived from the recombination of a human VH gene segment, human D gene segments and a human JH segment, wherein the VH gene segment

(a) comprises a nucleotide sequence that comprises a single nucleotide polymorphism rs56069819;

or

(b) is VH3-23*04 of SEQ ID NO;

38; and

wherein said human comprises;

(iv) a VH domain derived from the recombination of a human VH gene segment, human D gene segments and a human JH segment, wherein the VH gene segment is said VH gene segment of (iii); and

(v) said nucleotide sequence selected from the group consisting of SEQ ID NOs;

29-37.

2. The method of claim 1, comprising, before said administering, selecting a human comprising said nucleotide sequence of (v) or a polypeptide encoded by said nucleotide sequence of (v) and wherein the human is the human of claim 1.

3. The method of claim 1, wherein the antibody or antibody fragment specifically binds a PCSK9 comprising the amino acid sequence of SEQ ID NO:

3.

4. The method of claim 1, wherein the antibody or antibody fragment binds a second polypeptide comprising (vi) a PCSK9 amino acid sequence encoded by a PCSK9 nucleotide sequence selected from the group consisting of SEQ ID NOs:

29-37;

or (vii) an amino acid sequence comprising at least the catalytic domain or C-terminal domain of a PCSK9 protein encoded by any of SEQ ID NOs;

29-37.

5. The method of claim 1, wherein the human has been determined to comprise the nucleotide sequence of (v).

6. The method of claim 1, wherein the human has been determined to comprise a polypeptide encoded by the nucleotide sequence of (v).

7. The method of claim 1, comprising the step of determining that the human comprises the nucleotide sequence of (v).

8. The method of claim 7, wherein the determining step is performed before administration of the antibody to the human.

9. The method of claim 1, comprising the step of determining that the human comprises a polypeptide encoded by the nucleotide sequence of (v).

10. The method of claim 9, wherein the determining step is performed before administration of the antibody to the human.

11. The method of claim 8, wherein the step of determining comprises assaying a biological sample from the human for a nucleotide sequence of (v).

12. The method of claim 11, 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 said nucleotide sequence selected from the group consisting of SEQ ID NOs:

29-37 or at least the catalytic domain- or C-terminal domain-encoding sequence thereof, 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 selected from the group consisting of SEQ ID NOs;

29-37 or at least the catalytic domain- or C-terminal domain-encoding sequence thereof is present; and

/or

b. at least one oligonucleotide probe comprising a sequence of at least 10 contiguous nucleotides of said nucleotide sequence selected from the group consisting of SEQ ID NOs;

29-37 or 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 a nucleotide sequence selected from the group consisting of SEQ ID NOs;

29-37 is present; and

detecting the presence or absence of the complex, wherein detecting the presence of the complex determines that the human comprises said nucleotide sequence.

13. The method of claim 11, 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.

14. The method of claim 11, wherein the assaying is performed in a multiplex format.

15. The method of claim 11, further comprising obtaining the biological sample from the human.

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

17. The method of claim 16, wherein the human is receiving or has received statin treatment or has reduced responsiveness to statin treatment.

18. The method of claim 1, wherein the human is further administered a statin.

19. The method of claim 18, wherein said antibody or antibody fragment and said statin are administered separately or simultaneously.

20. The method of claim 11, wherein said biological sample comprises serum, blood, feces, tissue, a cell, urine and/or saliva of said human.

21. The method of claim 1, wherein said human is indicated as heterozygous for said nucleotide sequence of (v).

22. The method of claim 21, wherein said human is further indicated as comprising the nucleotide sequence of SEQ ID NO:

28 and/or the catalytic domain- or C-terminal domain-encoding sequence thereof.

23. The method of claim 1, wherein said human is indicated as homozygous for said nucleotide sequence of (v).

24. 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.

25. The method of claim 1, wherein said method treats or prevents 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.

26. The method of claim 1, wherein the method comprises administering alirocumab to the human.

27. The method of claim 1, wherein the nucleotide sequence selected from the group of (i) or (ii) is the nucleotide sequence SEQ ID NO:

30, SEQ ID NO;

31 or SEQ ID NO;

37.

28. The method of claim 17, wherein the method comprises administering alircoumab to the human.

29. The method of claim 27, wherein the method comprises administering alircoumab to the human.