Granted Patent B2

US 12,747,272 · App. 17/605,859 · Granted Sep 29, 2026

Inventors: Seth Carbonneau (Roxbury, MA); Marc Horst Peter Hild (Wellesley, MA); Andrei Golosov (Cambridge, MA); Nicole Renaud (Somerville, MA)

Assignee: Novartis AG

C07K14/4702A61K40/11A61K40/31A61K40/4211A61K40/4215C07K14/7051A61K2239/31A61K2239/38C07K2319/03C07K2319/95

Abstract

Claims (40)

1 . A fusion polypeptide comprising a degradation polypeptide and a heterologous polypeptide, wherein

the degradation polypeptide comprises the amino acid sequence of FQCEICGFSCR (SEQ ID NO: 1584) or FQCEICGASFR (SEQ ID NO: 1624).

2 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises

the amino acid sequence of FQCEICGASFRQKGNLLRHIKLH (SEQ ID NO: 1697).

3 . The fusion polypeptide of claim 1 , wherein:

the degradation polypeptide is between 15 and 90 amino acid residues in length, between 20 and 85 amino acid residues in length, between 25 and 80 amino acid residues in length, between 30 and 75 amino acid residues in length, between 35 and 70 amino acid residues in length, between 40 and 65 amino acid residues in length, between 45 and 65 amino acid residues in length, between 50 and 65 amino acid residues in length, or between 55 and 65 amino acid residues in length.

4 . The fusion polypeptide of claim 1 , wherein the heterologous polypeptide is a transmembrane polypeptide.

5 . The fusion polypeptide of claim 4 , wherein the transmembrane polypeptide is selected from the group consisting of CD62L, CCR1, CCR2, CCR5, CCR7, CCR10, CXCR2, CXCR3, CXCR4, CXCR6, CTLA4, PD1, BTLA, VISTA, CD137L, CD80, CD86, TIGIT, CD3, CD8, CD19, CD22, CD20, BCMA, and a chimeric antigen receptor (CAR).

6 . A pharmaceutical composition comprising the fusion polypeptide of claim 1 .

7 . The fusion polypeptide of claim 1 , wherein:

(i) in the presence of an immunomodulatory imide drug (IMiD), the expression level of the fusion polypeptide is decreased as compared to the expression level of the fusion polypeptide in the absence of the IMiD;

(ii) in the absence of an IMiD, the expression level of the fusion polypeptide is increased as compared to the expression level of an otherwise similar fusion polypeptide that comprises a degradation polypeptide comprising the amino acid sequence of FQCNQCGASFT (SEQ ID NO: 1561); or

(iii) in the presence of an IMiD, the expression level of the fusion polypeptide is decreased as compared to the expression level of an otherwise similar fusion polypeptide that comprises a degradation polypeptide comprising the amino acid sequence of FQCNQCGASFT (SEQ ID NO: 1561).

8 . The fusion polypeptide of claim 7 , wherein the IMiD is lenalidomide, pomalidomide, or thalidomide.

9 . The fusion polypeptide of claim 1 , wherein the heterologous polypeptide is a CAR comprising an antigen binding domain, a transmembrane domain, and an intracellular signaling domain.

10 . The fusion polypeptide of claim 9 , wherein the intracellular signaling domain comprises a costimulatory domain and a primary signaling domain, wherein:

(i) the degradation polypeptide is between the costimulatory domain and the primary signaling domain;

(ii) the fusion polypeptide comprises, from the N-terminus to the C-terminus, the antigen binding domain, the transmembrane domain, the costimulatory domain, the degradation polypeptide, and the primary signaling domain; or

(iii) the fusion polypeptide comprises, from the N-terminus to the C-terminus, the antigen binding domain, the transmembrane domain, a 4-1BB costimulatory domain, a first linker, the degradation polypeptide, a second linker, and a CD3-zeta stimulatory domain.

11 . The fusion polypeptide of claim 9 , wherein:

(i) the antigen binding domain binds an antigen selected from the group consisting of CD19; CD123; CD22; CD30; CD171; CS-1; C-type lectin-like molecule-1, CD33; epidermal growth factor receptor variant III (EGFRvIII); ganglioside G2 (GD2); ganglioside GD3; TNF receptor family member; B-cell maturation antigen; Tn antigen ((Tn Ag) or (GalNAca-Ser/Thr)); prostate-specific membrane antigen (PSMA); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Fms-Like Tyrosine Kinase 3 (FLT3); Tumor-associated glycoprotein 72 (TAG72); CD38; CD44v6; Carcinoembryonic antigen (CEA); Epithelial cell adhesion molecule (EPCAM); B7H3 (CD276); KIT (CD117); Interleukin-13 receptor subunit alpha-2; Mesothelin; Interleukin 11 receptor alpha (IL-11Ra); prostate stem cell antigen (PSCA); Protease Serine 21; vascular endothelial growth factor receptor 2 (VEGFR2); Lewis (Y) antigen; CD24; Platelet-derived growth factor receptor beta (PDGFR-beta); Stage-specific embryonic antigen-4 (SSEA-4); CD20; Folate receptor alpha; Receptor tyrosine-protein kinase ERBB2 (Her2/neu); Mucin 1, cell surface associated (MUC1); epidermal growth factor receptor (EGFR); neural cell adhesion molecule (NCAM); Prostase; prostatic acid phosphatase (PAP); elongation factor 2 mutated (ELF2M); Ephrin B2; fibroblast activation protein alpha (FAP); insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX); Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2); glycoprotein 100 (gp100); oncogene polypeptide consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl); tyrosinase; ephrin type-A receptor 2 (EphA2); Fucosyl GM1; sialyl Lewis adhesion molecule (sLe); ganglioside GM3; transglutaminase 5 (TGS5); high molecular weight-melanoma-associated antigen (HMWMAA); 0-acetyl-GD2 ganglioside (OAcGD2); Folate receptor beta; tumor endothelial marker 1 (TEM1/CD248); tumor endothelial marker 7-related (TEM7R); claudin 6 (CLDN6); thyroid stimulating hormone receptor (TSHR); G protein-coupled receptor class C group 5, member D (GPRC5D); chromosome X open reading frame 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK); Polysialic acid; placenta-specific 1 (PLAC1); hexasaccharide portion of globoH glycoceramide (GloboH); mammary gland differentiation antigen (NY-BR-1); uroplakin 2 (UPK2); Hepatitis A virus cellular receptor 1 (HAVCR1); adrenoceptor beta 3 (ADRB3); pannexin 3 (PANX3); G protein-coupled receptor 20 (GPR20); lymphocyte antigen 6 complex, locus K 9 (LY6K); Olfactory receptor 51E2 (OR51E2); TCR Gamma Alternate Reading Frame Protein (TARP); Wilms tumor protein (WT1); Cancer/testis antigen 1 (NY-ESO-1); Cancer/testis antigen 2 (LAGE-1a); Melanoma-associated antigen 1 (MAGE-A1); ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); X Antigen Family, Member 1A (XAGE1); angiopoietin-binding cell surface receptor 2 (Tie 2); melanoma cancer testis antigen-1 (MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); Fos-related antigen 1; tumor protein p53 (p53); p53 mutant; prostein; surviving; telomerase; prostate carcinoma tumor antigen-1, melanoma antigen recognized by T cells 1; Rat sarcoma (Ras) mutant; human Telomerase reverse transcriptase (hTERT); sarcoma translocation breakpoints; melanoma inhibitor of apoptosis (ML-IAP); ERG (transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene); N-Acetyl glucosaminyl-transferase V (NA17); paired box protein Pax-3 (PAX3); Androgen receptor; Cyclin B1; v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN); Ras Homolog Family Member C (RhoC); Tyrosinase-related protein 2 (TRP-2); Cytochrome P450 1B1 (CYP1B1); CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3); Paired box protein Pax-5 (PAX5); proacrosin binding protein sp32 (OY-TES1); lymphocyte-specific protein tyrosine kinase (LCK); A kinase anchor protein 4 (AKAP-4); synovial sarcoma, X breakpoint 2 (SSX2); Receptor for Advanced Glycation Endproducts (RAGE-1); renal ubiquitous 1 (RU1); renal ubiquitous 2 (RU2); legumain; human papilloma virus E6 (HPV E6); human papilloma virus E7 (HPV E7); intestinal carboxyl esterase; heat shock protein 70-2 mutated (mut hsp70-2); CD79a; CD79b; CD72; Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); Fc fragment of IgA receptor (FCAR or CD89); Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); CD300 molecule-like family member f (CD300LF); C-type lectin domain family 12 member A (CLEC12A); bone marrow stromal cell antigen 2 (BST2); EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2); lymphocyte antigen 75 (LY75); Glypican-3 (GPC3); Fc receptor-like 5 (FCRL5); BCMA; and immunoglobulin lambda-like polypeptide 1 (IGLL1);

(ii) the intracellular signaling domain comprises a primary signaling domain comprising a functional signaling domain derived from a protein selected from the group consisting of CD3-zeta, TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, CD278 (ICOS), FcεRI, DAP10, DAP12, and CD66d;

(iii) the intracellular signaling domain comprises a costimulatory domain comprising a functional signaling domain derived from a protein selected from the group consisting of MHC class I molecules, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, ICAM-1, 4-1BB (CD137), B7-H3, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83; or

(iv) the transmembrane domain comprises a transmembrane region of: the alpha, beta or zeta chain of the T-cell receptor, CD28, CD27, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154.

12 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises the amino acid sequence of HTGERPFQCEICGASFRQKGNLLRHIKLH (SEQ ID NO: 1699).

13 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises the amino acid sequence of FQCEICGFSCRQKGNLLRHIKLH (SEQ ID NO: 1698).

14 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises the amino acid sequence of HTGERPFQCEICGFSCRQKGNLLRHIKLH (SEQ ID NO: 1700).

15 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide further comprises the amino acid sequence of HKRSHTGERP (SEQ ID NO: 1694), HTGERP (SEQ ID NO: 1701), or GERP (SEQ ID NO: 1696).

16 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide further comprises the amino acid sequence of TGEKPFKCHLCN (SEQ ID NO: 1695).

17 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide further comprises the amino acid sequence of QKGNLLRHIKLH (SEQ ID NO: 1702).

18 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide further comprises the amino acid sequence of TASAEARHIKAEMG (SEQ ID NO: 11).

19 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises the amino acid sequence of SEQ ID NO: 2064 or SEQ ID NO: 2231.

20 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises the amino acid sequence of SEQ ID NO: 2140.

21 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide comprises the amino acid sequence of any of SEQ ID NOs.: 2137-2139, 2141, or 2142.

22 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide and the heterologous polypeptide are linked by a peptide bond.

23 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide and the heterologous polypeptide are linked by a bond other than a peptide bond.

24 . The fusion polypeptide of claim 1 , wherein the heterologous polypeptide is linked directly to the degradation polypeptide.

25 . The fusion polypeptide of claim 1 , wherein the heterologous polypeptide is linked indirectly to the degradation polypeptide.

26 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide and the heterologous polypeptide are operatively linked via a linker.

27 . The fusion polypeptide of claim 1 , wherein the degradation polypeptide is linked to the C-terminus or N-terminus of the heterologous polypeptide.

Assignments (4)

Continuity (2)

References Cited (400)