Huwentoxin-IV variants and methods of use

• US 9,102,751 B2
• Filed: 03/15/2013
• Issued: 08/11/2015
• Est. Priority Date: 05/18/2012
• Status: Active Grant

• Current Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Original Assignee: Janssen Biotech, Inc. (Johnson & Johnson)
• Inventors: Flinspach, Mack, Hunter, Michael, Liu, Yi, Neff, Robert, Wickenden, Alan, Gibbs, Alan
• Primary Examiner(s): Cordero Garcia, Marcela M
• Assistant Examiner(s): Coffa, Sergio
• Application Number:
  US13/833,555
• Publication Number:
  US 20130310324A1
• Time in Patent Office: 879 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 23/00 Anaesthetics
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: C07K 14/43518 from spiders

Abstract

The present invention relates to Huwentoxin-IV variants, polynucleotides encoding them, and methods of making and using the foregoing.

Claims

1. An isolated non-natural Huwentoxin-IV variant comprising the sequence X₁CX₂X₃X₄FX₅X₆CX₇X₈X₉X₁₀X₁₁X₁₂CCX₁₃X₁₄ X₁₅X₁₆X₁₇X₁₈CX₁₉X₂₀X₂₁X₂₂X₂₃ X₂₄CKX₂₅X₂₆IX₂₇X₂₈ (SEQ ID NO:

357);

wherein

i) X₁ is K, R, H, D, Y, F, N, Q, S, T, G, L, I, P or E;

ii) X₂ is R, F, W, N, S or L;

iii) X₃ is R, H, D, Y, N, Q, L, I, P or E;

iv) X₄ is Y, V or I;

v) X₅ is R, W, Q, S or K;

vi) X₆ is R, E, Y, F, V or A;

vii) X₇ is K, R, E, Y, F, S, V or N;

viii) X₈ is P or V;

ix) X₉ is R, F, Q, V or S;

x) X₁₀ is H, D, Y, W, Q, S, T, G, A, V, L, I, P or N;

xi) X₁₁ is D, P or W;

xii) X₁₂ is K, R, D, E, Y, W, N, T, A, L or Q;

xiii) X₁₃ is R, Y, Q, S, T, G, L, I, P or K;

xiv) X₁₄ is K, R, Y, F, N, Q, G, A, V, L, I, P or S;

xv) X₁₅ is R, H, D, Y, W, N, Q, T, V, I, P or S;

xvi) X₁₆ is R, H, D, F, W, N, Q, S, T, G, A, L or K;

xvii) X₁₇ is K, R, Y, F, W, P or L;

xviii) X₁₈ is K, R, T, A, L or V;

xix) X₁₉ is S or I;

xx) X₂₀ is K, W, G, A, I, R or D;

xxi) X₂₁ is Y, W, A, H or K;

xxii) X₂₂ is T or V;

xxiii) X₂₃ is K, H, W, N, G, A, L or R;

xxiv) X₂₄ is W or K;

xxv) X₂₅ is W, T, I or Y;

xxvi) X₂₆ is K, R, Y, F, S, T, G, A, V, L, I or Q;

xxvii) X₂₇ is K, R, H, F, W, V, L, I, G or deleted; and

xxviii) X₂₈ is R, H, Y, F, W, N, G, V, P, K or deleted, and

the non-natural Huwentoxin-IV variant has an IC₅₀ value of about 300×

10<sup>−

9</sup> M or less for human Nav1.7 (SEQ ID NO;

     263), with the proviso that the non-natural Huwentoxin-IV variant is not a polypeptide comprising the sequence shown in SEQ ID NO;

1.

2. The isolated non-natural Huwentoxin-IV variant of claim 1, wherein the variant comprises the sequence X₁CX₂X₃X₄FX₅X₆CX₇X₈X₉X₁₀X₁₁X₁₂CCX₁₃X₁₄X₁₅X₁₆X₁₇X₁₈CX₁₉X₂₀X₂₁TX₂₂ WCKYX₂₃IX₂₄X_{25 (SEQ ID NO:}

359);

wherein

i) X₁ is K, R, H, D, Y, F, N, Q, S, T, G, L, I, P or E;

ii) X₂ is R, F, W, N, S or L;

iii) X₃ is R, H, D, Y, N, Q, L, I, P or E;

iv) X₄ is Y, V or I;

v) X₅ is R, W, Q, S or K;

vi) X₆ is R, E, Y, F, V or A;

vii) X₇ is K, R, E, Y, F, S, V or N;

viii) X₈ is P or V;

ix) X₉ is R, F, Q, V or S;

x) X₁₀ is H, D, Y, W, Q, S, T, G, A, V, L, I, P or N;

xi) X₁₁ is D, P or W;

xii) X₁₂ is K, R, D, E, Y, W, N, T, A, L or Q;

xiii) X₁₃ is R, Y, Q, S, T, G, L, I, P or K;

xiv) X₁₄ is K, R, Y, F, N, Q, G, A, V, L, I, P or S;

xv) X₁₅ is R, H, D, Y, W, N, Q, T, V, I, P or S;

xvi) X₁₆ is R, H, D, F, W, N, Q, S, T, G, A, L or K;

xvii) X₁₇ is K, R, Y, F, W, P or L;

xviii) X₁₈ is K, R, T, A, L or V;

xix) X₁₉ is S or I;

xx) X₂₀ is K, W, G, A, I, D or R;

xxi) X₂₁ is Y, W, A, H or K;

xxii) X₂₂ is K, H, W, N, G, A, L or R;

xxiii) X₂₃ is K, R, Y, F, S, T, G, A, V, L, I or Q;

xxiv) X₂₄ is K, R, H, F, W, V, L, I, G or deleted; and

xxv) X₂₅ is R, H, Y, F, W, N, G, V, P, K or deleted.

3. The isolated non-natural Huwentoxin-IV variant of claim 1, wherein the non-natural Huwentoxin-IV variant has IC₅₀ of less than about 160×

10<sup>−

9</sup> for human Nav1.7.

4. The isolated non-natural Huwentoxin-IV variant of claim 3, wherein the non-natural Huwentoxin-IV variant comprises the polypeptide sequence of SEQ ID NOs:

3-222, 241-253 or 227-280.

5. The isolated non-natural Huwentoxin-IV variant of claim 1, wherein the non-natural Huwentoxin-IV variant selectively inhibits Nav1.7 and has a ration of IC₅₀ for Nav1.2 to IC₅₀ for Nav1.7 (Nav1.2)/IC₅₀ (Nav1.7)) equal or over about 5.0.

7. A pharmaceutical composition comprising the isolated non-natural Huwentoxin-IV variant of claim 1 and a pharmaceutically acceptable excipient.

9. The isolated non-natural Huwentoxin-IV variant of claim 5, wherein the non-natural Huwentoxin-IV variant comprises the polypeptide sequence of SEQ ID NOs:

5, 7, 12, 13, 16, 21, 25, 45, 46, 48, 55, 57, 58, 60, 51, 72, 74, 76, 78, 82, 83, 96, 109, 111, 113, 122, 127, 131, 134, 137, 141, 142, 149, 164, 165, 172, 175, 177, 178, 180, 182, 188, 189, 192, 198, 202, 204, 213, 215, 219, 223-240 or 277.

10. An isolated fusion protein comprising the non-natural Huwentoxin-IV variant of claim 1 fused with a second poypeptide.

6. An isolated non-natural Huwentoxin-IV variant comprising the polypeptide sequence shown in SEQ ID NOs:

3-253 or 277-355.

8. A method of treating pain in a subject, comprising administering to the subject an effective amount of the Huwentoxin-IV variant of claim 6 to treat the pain.