SILVESTROL ANTIBODY-DRUG CONJUGATES AND METHODS OF USE

• US 20170348422A1
• Filed: 06/05/2017
• Published: 12/07/2017
• Est. Priority Date: 06/06/2016
• Status: Active Grant

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Pillow, Thomas, Polson, Andrew G., Zheng, Bing
• Granted Patent: US 10,639,378 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/572 cytotoxic response
• CPC Class Codes: A61K 2039/627 characterised by the linker
• CPC Class Codes: A61K 31/357 having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/541 Organic ions forming an ion pair complex with the pharmacologically or therapeutically active agent
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6855 the tumour determinant being from breast cancer cell
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07D 407/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: G01N 33/567 utilising isolate of tissue or organ as binding agent

Abstract

The invention relates generally to a silvestrol molecule activated with a leaving group. The invention further relates generally to an antibody-drug conjugate comprising an antibody conjugated by a linker to one or more silvestrol drug moieties and methods of treatment.

Claims

1. An antibody-drug conjugate compound comprising an antibody covalently attached through a linker to a silvestrol drug moiety, selected from Formulas Ia and Ib:

4. The antibody-drug conjugate compound according to claim 1, wherein Ab is a cysteine-engineered antibody.

5. The antibody-drug conjugate compound according to claim 4, wherein the cysteine-engineered antibody is a mutant selected from HC A118C, LC K149C, HC A140C, LC V205C, LC S121C, and HC L177C.

6. The antibody-drug conjugate compound according to claim 1 wherein L is a protease-cleavable, non-peptide linker having the formula:

-Str-PM-Y—

where Str is a stretcher unit covalently attached to the antibody;

PM is a peptidomimetic unit, and Y is a spacer unit covalently attached to the silvestrol drug moiety.

7. The antibody-drug conjugate compound according to claim 6, wherein Str has the formula:

8. The antibody-drug conjugate compound of claim 7 wherein R⁶ is (CH₂)₅.

9. The antibody-drug conjugate compound of claim 6 wherein PM has the formula:

10. The antibody-drug conjugate compound of claim 6 wherein Y comprises para-aminobenzyl or para-aminobenzyloxycarbonyl.

11. The antibody-drug conjugate compound of claim 9 having the formula:

12. The antibody-drug conjugate compound of claim 11 having the formula:

13. The antibody-drug conjugate compound of claim 11 having the formula:

14. The antibody-drug conjugate compound of claim 13 having the formula:

15. The antibody-drug conjugate compound of claim 14 selected from the formulas:

16. The antibody-drug conjugate compound according to claim 1 wherein L is a peptide linker having the formula:

-Str-Pep-Y—

where Str is a stretcher unit covalently attached to the antibody;

Pep is a peptide of two to twelve amino acid residues, and Y is a spacer unit covalently attached to the silvestrol drug moiety.

17. The antibody-drug conjugate compound according to claim 16, wherein Str has the formula:

18. The antibody-drug conjugate compound of claim 17 wherein R⁶ is (CH₂)₅ .

19. The antibody-drug conjugate compound of claim 16, wherein Pep comprises two to twelve amino acid residues independently selected from glycine, alanine, phenylalanine, lysine, arginine, valine, and citrulline.

20. The antibody-drug conjugate compound of claim 19, wherein Pep is selected from valine-citrulline, alanine-phenylalanine, and phenylalanine-lysine.

21. The antibody-drug conjugate compound of claim 16, wherein Y comprises para-aminobenzyl or para-aminobenzyloxycarbonyl.

22. The antibody-drug conjugate compound of claim 19 having the formula:

23. The antibody-drug conjugate compound of claim 22 having the formula:

24. The antibody-drug conjugate compound of claim 22 having the formula:

25. The antibody-drug conjugate compound of claim 24 having the formula:

26. The antibody-antibiotic conjugate of claim 22 having the formula:

27. The antibody-antibiotic conjugate of claim 26 having the formula:

28. The antibody-antibiotic conjugate of claim 22 having the formula:

29. The antibody-antibiotic conjugate of claim 28 having the formula:

30. The antibody-drug conjugate compound according to claim 1, wherein L comprises a disulfide group.

31. The antibody-drug conjugate compound according to claim 1, wherein p is 1, 2, 3, or 4.

32. The antibody-drug conjugate compound according to claim 1, comprising a mixture of the antibody-drug conjugate compounds, wherein the average drug loading per antibody in the mixture of antibody-drug conjugate compounds is about 2 to about 5.

33. A pharmaceutical composition comprising the antibody-drug conjugate compound according to claim 1 and a pharmaceutically acceptable diluent, carrier or excipient.

34. A method of treating cancer comprising administering to a patient a therapeutically effective amount of the pharmaceutical composition of claim 33.

35. The method of claim 34 wherein the patient is administered a chemotherapeutic agent, in combination with the pharmaceutical composition comprising the antibody-drug conjugate compound.

36. An article of manufacture comprising a pharmaceutical composition of claim 33, a container, and a package insert or label indicating that the pharmaceutical composition can be used to treat cancer.

53. A method of making an antibody-drug conjugate compound according to claim 1, the method comprising reacting an antibody with a silvestrol-linker intermediate compound of Formula II of claim 37.

2. The antibody-drug conjugate compound of claiml wherein the antibody which binds to one or more tumor-associated antigens or cell-surface receptors is selected from (1)-(53):

(1) BMPR1B (bone morphogenetic protein receptor-type IB);

(2) E16 (LAT1, SLC7A5);

(3) STEAP1 (six transmembrane epithelial antigen of prostate);

(4) MUC16 (0772P, CA125);

(5) MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin);

(6) Napi2b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34 (sodium phosphate), member 2, type II sodium-dependent phosphate transporter 3b);

(7) Sema 5b (F1110372, KIAA1445, Mm.42015, SEMA5B, SEMAG, Semaphorin 5b Hlog, sema domain, seven thrombospondin repeats (type 1 and type 1-like), transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 5B);

(8) PSCA hlg (2700050C12Rik, C530008O16Rik, RIKEN cDNA 2700050C12, RIKEN cDNA 2700050C12 gene);

(9) ETBR (Endothelin type B receptor);

(10) MSG783 (RNF124, hypothetical protein F1120315);

(11) STEAP2 (HGNC_8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostate cancer associated gene 1, prostate cancer associated protein 1, six transmembrane epithelial antigen of prostate 2, six transmembrane prostate protein);

(12) TrpM4 (BR22450, F1120041, TRPM4, TRPM4B, transient receptor potential cation channel, subfamily M, member

     4);

(13) CRIPTO (CR, CR1, CRGF, CRIPTO, TDGF1, teratocarcinoma-derived growth factor);

(14) CD21 (CR2 (Complement receptor

     2) or C3DR (C3d/Epstein Barr virus receptor) or Hs

     73792);

(15) CD79b (CD79B, CD7913, IGb (immunoglobulin-associated beta), B29);

(16) FcRH2 (IFGP4, IRTA4, SPAP1A (SH2 domain containing phosphatase anchor protein 1a), SPAP1B, SPAP1C);

(17) HER2;

(18) NCA;

(19) MDP;

(20) IL20Rα

;

(21) Brevican;

(22) EphB2R;

(23) ASLG659;

(24) PSCA;

(25) GEDA;

(26) BAFF-R (B cell -activating factor receptor, BLyS receptor 3, BR3);

(27) CD22 (B-cell receptor CD22-B isoform);

(28) CD79a (CD79A, CD79α

, immunoglobulin-associated alpha);

(29) CXCR5 (Burkitt'"'"'s lymphoma receptor

     1);

(30) HLA-DOB (Beta subunit of MEW class II molecule (Ia antigen));

(31) P2X5 (Purinergic receptor P2X ligand-gated ion channel

     5);

(32) CD72 (B-cell differentiation antigen CD72, Lyb-2);

(33) LY64 (Lymphocyte antigen 64 (RP105), type I membrane protein of the leucine rich repeat (LRR) family);

(34) FcRH1 (Fc receptor-like protein

     1);

(35) FcRH5 (IRTA2, Immunoglobulin superfamily receptor translocation associated

     2);

(36) TENB2 (putative transmembrane proteoglycan);

(37) PMEL17 (silver homolog;

SILV;

D12S53E;

PMEL17;

SI;

SIL);

(38) TMEFF1 (transmembrane protein with EGF-like and two follistatin-like domains 1;

Tomoregulin-1);

(39) GDNF-Ra1 (GDNIF family receptor alpha 1;

GFRA1;

GDNFR;

GDNFRA;

RETL1;

TRNR1;

RET1L;

GDNFR-alphal;

GFR-ALPHA-1);

(40) Ly6E (lymphocyte antigen 6 complex, locus E;

Ly67,RIG-E,SCA-2,TSA-1);

(41) TMEM46 (shisa homolog 2 (Xenopus laevis);

SHISA2);

(42) Ly6G6D (lymphocyte antigen 6 complex, locus G6D;

Ly6-D, MEGT1);

(43) LGRS (leucine-rich repeat-containing Ci protein-coupled receptor 5;

GPR49, GPR67);

(44) RET (ret proto-oncogene;

MEN2A;

HSCR1;

MEN2B;

MTC1;

PTC;

CDHF12;

Hs.168114;

RET51;

RET-ELE1);

(45) LY6K (lymphocyte antigen 6 complex, locus K;

LY6K;

HSJ001348;

FLJ35226);

(46) GPR19 (G protein-coupled receptor 19;

Mm.4787);

(47) GPR54 (KISS1 receptor;

KISS1R;

GPR54;

HOT7T175;

AXOR12);

(48) ASPHD1 (aspartate beta-hydroxylase domain containing 1;

LOC253982);

(49) Tyrosinase (TYR;

OCAIA;

OCA1A;

tyrosinase;

SHEP3);

(50) TMEM118 (ring finger protein, transmembrane 2;

RNFT2;

FLJ14627);

(51) GPR172A (G protein-coupled receptor 172A;

GPCR41;

FLJ11856;

D15Ertd747e);

(52) CD33; and

(53) CLL-1.

3. The antibody-drug conjugate compound according to claim 2, wherein Ab is selected from anti-HER2 4D5, anti-CD22, anti-CD33, anti- Ly6E, anti-Napi3b, anti-HER2 7C2, and anti-CLL-1.

37. A silvestrol-linker intermediate compound of Formula II:

38. The silvestrol-linker intermediate compound of claim 37 wherein L-X is selected from —

CH₂CH₂—

X, —

CH₂CR₂—

X, —

C(O)NRCH₂—

X, —

CH₂O—

X, —

CH₂N(R)—

X, —

N(R)—

X, —

N(R)(C₁-C₁₂ alkylene)-X, —

N(R)(C₂-C₈ alkenylene)-X, —

N(R)(C₂-C₈ alkynylene)-X, and —

N(R)(CH₂CH₂O)_n—

X;

n is 1 to 6;

R is independently selected from H, C₁-C₁₂ alkyl, and C₆-C₂₀ aryl;

or two R form a C₃-C₇ carbocyclic ring; and

alkylene, alkenylene, alkynylene, alkyl, and aryl are optionally substituted with one or more groups selected from F, Cl, Br, N(CH₃)₂, NO₂, and OCH₃.

39. The silvestrol-linker intermediate compound of claim 38, wherein alkylene is selected from —

CH₂CH₂—

, —

CH₂CH₂CH₂—

, —

CH(CH₃)CH₂—

, and —

C(CH₃)₂CH₂—

.

40. The silvestrol-linker intermediate compound of claim 37 wherein X is selected from:

41. The silvestrol-linker intermediate compound of claim 37 wherein R^a is —

OCH₃.

42. The silvestrol-linker intermediate compound of claim 37 wherein R¹ is —

OCH₃ and R² is L-X.

43. The silvestrol-linker intermediate compound of claim 37 wherein R¹ is L-X and R² is —

CH(OH)CH₂OH.

44. The silvestrol-linker intermediate compound of claim 37 wherein L is a protease-cleavable, non-peptide linker having the formula:

-Str-PM-Y—

where Str is a stretcher unit covalently attached to X;

PM is a peptidomimetic unit, and Y is a spacer unit covalently attached to the silvestrol drug moiety.

45. The silvestrol-linker intermediate compound of claim 44 wherein Str is (CH₂)₅.

46. The silvestrol-linker intermediate compound of claim 44 wherein PM has the formula:

47. The silvestrol-linker intermediate compound of claim 46 wherein R⁷ and R⁸ together form cyclobutyl.

48. The silvestrol-linker intermediate compound of claim 44 wherein Y comprises para-aminobenzyl or para-aminobenzyloxycarbonyl.

49. The silvestrol-linker intermediate compound of claim 37 wherein L is a peptide linker having the formula:

-Str-Pep-Y—

where Str is a stretcher unit covalently attached to the antibody;

Pep is a peptide of two to twelve amino acid residues, and Y is a spacer unit covalently attached to the silvestrol drug moiety.

50. The silvestrol-linker intermediate compound of claim 49 wherein Str is (CH₂)₅.

51. The silvestrol-linker intermediate compound of claim 49 wherein Pep comprises two amino acid residues independently selected from glycine, alanine, phenylalanine, lysine, arginine, valine, and citrulline.

52. The silvestrol-linker intermediate compound of claim 37 selected from: