Pseudotyped viral particles, compositions comprising the same, and uses thereof

Abstract

Provided herein are variant VSV-G polypeptides comprising one or more mutations as compared to wild-type VSV-G polypeptides, compositions comprising the variant VSV-G polypeptides, and methods of using the same.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480047724.3 (22) Application Date 2024.06.11 (30) Priority Data 63 / 507,550 2023.06.12 US (85) PCT International Application Entering National Phase Date 2026.01.16 (86) PCT International Application Application Data PCT / US2024 / 033426 2024.06.11 (87) PCT International Application Publication Data WO2024 / 258863 EN 2024.12.19 (71) Applicant Interus Biotherapeutics Company Address Pennsylvania, USA (72) Inventors S.J. Doolittle R.M. Russell (74) Patent Agency Beijing Liu Shen Law Firm 11105 Patent Attorney Qin Jian (51) Int.Cl. C07K 14 / 145 (2006.01) C12N 15 / 86 (2006.01) A61P 35 / 00 (2006.01) (54) Invention Title: Pseudotyped Viral Particles, Compositions Containing Pseudotyped Viral Particles and Use Thereof (57) Abstract: This document provides a variant VSV-G polypeptide containing one or more mutations compared to the wild-type VSV-G polypeptide, a composition containing the variant VSV-G polypeptide, and methods of using the present invention. Claims: 5 pages Description: 141 pages Sequence Listing (Electronic Publication) Drawings: 37 pages CN 121794286 A 2026.04.03 CN 1 21 79 42 86 A

Claims

1. A variant VSV-G polypeptide comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 182, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof in SEQ ID NO: 2, provided that the mutation is not an I182E mutation, an I182A mutation or an I182D mutation.

2. The variant VSV-G polypeptide according to claim 1, wherein the mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 182, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof in SEQ ID NO: 2 comprises a substitution to a charged amino acid.

3. The variant VSV-G polypeptide according to claim 2, wherein the charged amino acids correspond to the positions H8, N9, Q10, K47, K50, A51, S179, N180, I182, S183, M184, Y209, I347, T350, T352, E353 or R354 of SEQ ID NO:

2.

4. The variant VSV-G polypeptide according to claim 2, wherein the substitution of the charged amino acid is D, E, K or R.

5. The variant VSV-G polypeptide according to claim 2, wherein the mutation at position H8 toward the charged amino acid is H8D, H8E, H8K, or H8R.

6. The variant VSV-G polypeptide of claim 2, wherein the mutation at position N9 toward the charged amino acid is N9D, N9E, N9K, or N9R.

7. The variant VSV-G polypeptide according to claim 2, wherein the mutation at position Q10 toward the charged amino acid is Q10D or Q10E.

8. The variant VSV-G polypeptide according to claim 2, wherein the mutation at position K47 toward the charged amino acid is K47D or K47E.

9. The variant VSV-G polypeptide of claim 2, wherein the mutation at position K50 toward the charged amino acid is K50D or K50E.

10. The variant VSV-G polypeptide of claim 2, wherein the mutation at position A51 toward the charged amino acid is A51D, A51E, A51K, or A51R.

11. The variant VSV-G polypeptide according to claim 2, wherein the mutation at position S179 toward the charged amino acid is S179D, S179E, S179K or S179R.

12. The variant VSV-G polypeptide of claim 2, wherein the mutation at position N180 toward the charged amino acid is N180D, N180E, or N180K.

13. The variant VSV-G polypeptide of claim 2, wherein the mutation at position I182 toward the charged amino acid is I182D, I182K, or I182R.

14. The variant VSV-G polypeptide of claim 2, wherein the mutation at position S183 toward the charged amino acid is S183D, S183E, S183K or S183R.

15. The variant VSV-G polypeptide of claim 2, wherein the mutation at position M184 toward the charged amino acid is M184D, M184E, M184K, or M184R.

16. The variant VSV-G polypeptide of claim 2, wherein the mutation at position Y209 toward the charged amino acid is Y209D, Y209E, Y209K or Y209R.

17. The variant VSV-G polypeptide of claim 2, wherein the mutation at position I347 toward the charged amino acid is I347D, I347E, I347K, or I347R.

18. The variant VSV-G polypeptide of claim 2, wherein the mutation at position T350 toward the charged amino acid is T350D, T350E, T350K, or T350R.

19. The variant VSV-G polypeptide of claim 2, wherein the mutation at position T352 toward the charged amino acid is T352D, T352E, T352K or T352R.

20. The variant VSV-G polypeptide of claim 2, wherein the mutation at position E353 toward the charged amino acid is E353K or E353R.

21. The variant VSV-G polypeptide of claim 2, wherein the mutation at position R354 toward the charged amino acid is R354D or R354E.

22. The variant VSV-G polypeptide of claim 1, wherein the mutation is a substitution for alanine (A), glycine (G), glutamine (Q), or a combination thereof.

23. A variant VSV-G polypeptide comprising a variant amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5, the variant amino acid sequence comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof in SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO:

5.

24. The variant VSV-G polypeptide of claim 23, wherein the variant VSV-G polypeptide comprises a variant of the amino acid sequence of SEQ ID NO:3, the variant comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof in SEQ ID NO:

3.

25. The variant VSV-G polypeptide of claim 23, wherein the variant VSV-G polypeptide comprises a variant of the amino acid sequence of SEQ ID NO:4, the variant comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof of SEQ ID NO:

4.

26. The variant VSV-G polypeptide of claim 23, wherein the variant VSV-G polypeptide comprises a variant of the amino acid sequence of SEQ ID NO:5, the variant comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof in SEQ ID NO:

5.

27. The variant VSV-G polypeptide according to any one of claims 23 to 26, wherein the mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 182, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof comprises a substitution to a charged amino acid selected from the group consisting of H8, N9, Q10, K47, K50, A51, S179, N180, S183, M184, Y209, I347, T350, T352, E353 or R354.

28. A variant VSV-G polypeptide comprising a variant of the amino acid sequence of SEQ ID NO: 22 or SEQ ID NO: 23, the variant comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 353, 354 or combinations thereof in SEQ ID NO: 22 or SEQ ID NO:

23.

29. The variant VSV-G polypeptide of claim 28, wherein the variant VSV-G polypeptide comprises a variant of the amino acid sequence of SEQ ID NO:22, the variant comprising a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 353, 354 or combinations thereof of SEQ ID NO:

22.

30. The variant VSV-G polypeptide of claim 28, wherein the variant VSV-G polypeptide comprises the amino acid sequence of SEQ ID NO:23, the amino acid sequence comprising mutations corresponding to mutations at positions 8, 9, 10, 47, 50, 51, 179, 180, 183, 184, 209, 347, 350, 353, 354 or combinations thereof in SEQ ID NO:

23.

31. The variant VSV-G polypeptide according to any one of claims 28 to 30, wherein the mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 182, 183, 184, 209, 347, 350, 352, 353, 354 or combinations thereof comprises a substitution to a charged amino acid selected from the group consisting of H8, N9, Q10, K47, K50, A51, S179, N180, S183, M184, Y209, I347, T350, E353 or R354.

32. A variant VSV-G polypeptide, wherein the variant amino acid sequence has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence of SEQ ID NO: 2, and further comprises a mutation corresponding to a mutation at positions 8, 9, 10, 47, 50, 51, 179, 180, 182, 183, 184, 209, 347, 350, 352, 353, 354, or any combination thereof, compared to SEQ ID NO:

2.

33. A nucleic acid molecule encoding a variant VSV-G polypeptide according to any one of claims 1 to 32.

34. A viral particle comprising a polypeptide according to any one of claims 1 to 32.

35. The viral particle of claim 36, wherein the viral particle further comprises a targeting portion.

36. The viral particle according to claim 34 or 35, wherein the viral particle is a pseudotyped lentivirus.

37. The viral particle according to any one of claims 34 to 36, wherein the viral particle further comprises a nucleic acid molecule encoding a heterologous molecule of interest.

38. The viral particle of claim 37, wherein the heterologous molecule of interest is a chimeric antigen receptor ("CAR").

39. The viral particle according to any one of claims 34 to 38, wherein the targeting portion binds to: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCRα, TCRβ, TCRγ, TCRδ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7 or CXCR3, a glycosylated CD43 epitope expressed on acute leukemia or lymphoma but not on hematopoietic progenitor cells, a glycosylated CD43 epitope expressed on non-hematopoietic cancers, kinase-anchored protein 4 (AKAP-4), adrenaline receptor β3 (ADRB3), AFP, anaplastic lymphoma kinase (ALK), androgen receptor, angiopoietin-binding cell surface receptor 2 (TIP). 2) Anti-desmosome 1 (Dsgl) autoantibody, anti-desmosome 3 (Dsg3) autoantibody, B7H3 (CD276), biotin, bone marrow stromal cell antigen 2 (BST2), BST1 / CD157; cancer / testis antigen 1 (NY-ESO-1), cancer / testis antigen 2 (LAGE-la), carbonic anhydrase IX (CAIX), carcinoembryonic antigen (CEA), CCCTC binding factor (zinc finger protein)-like (BORIS or imprinted site regulator sibling factor), CCR4, CD5, CD19, CD20, CD22, CD24, CD30, CD32 (FCG) R2A), CD33, CD34, CD38, CD44v6, CD72, CD79a, CD79b, CD97, CD99, CD123, CD171, CD179a, CD179b-IGLll, CD200R, CD276 / B7H3, CD300 molecular-like family member f (CD300LF), CDH1-CD324, CDH6, CDH17, CDH19, chromosome X open reading frame 61 (CXORF61), sealing protein 6 (CLDN6), sealing protein 18.2 (CLD18A2 or CLDN18A.2), CMV pp65, C-MYC epitope tag, Cripto, CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24), CSF2RA (GM-CSFR-α), C-type lectin domain family 12 member A (CLEC12A), C-type lectin-like molecule-1 (CLL-1 or CLECL1), cyclin B1, cytochrome P450 IB 1 (CYP1B)1) DLL3, EBV-EBNA3c, EGF-bke module 2 containing mucin-like hormone receptor-like receptor (EMR2), Elongation factor 2 mutant (ELF2M), liver glycoside B2, liver glycoside A receptor 2 (EphA2), epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRviii), epidermal cell adhesion molecule (EPCAM), ERG, ETS translocation variant gene 6 located on chromosome 12p (ETV6-AML), Fc fragment of IgA receptor (FCAR or CD89), Fc receptor-like 5 (FCRL5), fibroblast activating protein α (FAP), FITC, Fms-like tyrosine kinase 3 (FLT3), folate receptor α (FRa or FR1), folate receptor β (FRb), follicle-stimulating hormone receptor (FSHR) Fos-related antigen 1, fucose-GM1, G protein-coupled receptor class C5 member D (GPRC5D), G protein-coupled receptor 20 (GPR20), GAD, ganglioside G2 (GD2), ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(11)Cer), ganglioside GM3 (aNeu5Ac(2-3)bDClalp(1-4)bDGlcp(11)Cer), GD3, GFRα4, glycoprotein 100 (gplOO), phosphatidylinositol proteoglycan-3 (GPC3), gonadotropin receptor (CGHR or GR), GpA33, GpNMB, GPRC5D, guanylate cyclase C (GCC), mutant heat shock protein 70-2 (mut HSP70-2), Hepatitis A Virus Receptor 1 (HAVCR1), the hexasaccharide moiety of globoH glycosylceramide (GloboH), high molecular weight melanoma-associated antigen (HMWMAA), HIV1 envelope glycoprotein, HLA, HLA-DOA, HLA-A, HLA-A2, HLA-B, HLA-C, HLA-DM, HLA-DOB, HLA-DP, HLA-DQ, HLA-DR, HLA-G, HTLV1-Tax, Human Papillomavirus E6 (HPV E6), Human Papillomavirus E7 (HPV E7), Human Telomerase Reverse Transcriptase (hTERT), IgE, IL13Ra2, IL1 lRa, immunoglobulin λ-like polypeptide 1 (IGLL1), influenza A hemagglutinin (HA), insulin-like growth factor 1 receptor (IGF-I receptor), interleukin 11 receptor α (IL-11Ra), interleukin 13 receptor subunit α-2 (IL-13Ra2 or CD213A2), intestinal carboxylesterase, KIT (CD117), KSHVK8.1, KSHV-gH, LAMP1, pod protein, leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), leukocyte-associated immunoglobulin-like receptor 1 (LAIR1), luteinizing hormone receptor (LHR), Lewis (Y) antigen, Lews Ag, Livl, locus K 9 (LY6K), low-conductivity chloride channels, lymphocyte antigen 6 complex, lymphocyte antigen 75 (LY75), lymphocyte-specific protein tyrosine kinase (LCK), breast differentiation antigen (NY-BR-1), T-cell recognized melanoma antigen 1 (MelanA or MARTI), melanoma-associated antigen 1 (MAGE-A1), melanoma cancer testis antigen-1 (MAD-CT-1), melanoma cancer testis antigen-2 (MAD-CT-2), melanoma cell apoptosis inhibitor (ML-IAP), mesothelin, MPL, cell surface-associated mucin 1 (MUC1), N-acetylglucosamine transferase V (NA17), adhesion protein-4, neural cell adhesion molecule (NCAM), NK G2D, NYBR1, o-acetyl-GD2 ganglioside (OAcGD2), olfactory receptor 51E2 (OR51E2), oncogene fusion protein composed of breakpoint cluster region (BCR) and Abelson murine leukemia virus oncogene homolog 1 (Abl) (bcr-abl), P53 mutant, pairing box protein Pax-3 (PAX3), pairing box protein Pax-5 (PAX5), pan-connector protein 3 (PANX3), PDL1, P-glycoprotein, placenta-specific 1 (PLAC1), platelet-derived growth factor receptor β (PDGFR-β), polysialic acid, proapocrine-binding protein sp32 (OY-TES1), prostate enzyme, prostate cancer tumor antigen-1 (PCT) Al or galactohemagglutinin 8), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), prostate acid phosphatase (PAP), Prostein, serine protease 21 (Testisin or PRSS21), proteasome macropain subunit βType 9 (LMP2), PTK7, RasG12V, Ras homolog family member C (RhoC), Rat sarcoma (Ras) mutant, receptor for advanced glycation end products (RAGE-1), receptor tyrosine kinase-like orphan receptor 1 (ROR1), receptor tyrosine protein kinase ERBB2 or Her-22 / neu, renal universal protein 1 (RU1), renal universal protein 2 (RU2), sarcoma translocation breakpoint, serine 2 (TMPRSS2) ETS fusion gene, sialic acid Lewis adhesion molecule (sLe), SLAMF4, SLAMF6, Slea (CA19.9 or sialic acid Lewis antigen), spermin 17 (SPA17), T cell recognized squamous cell carcinoma antigen 3 (SART3), stage-specific embryonic antigen-4 (SSEA-4), STEAP1, survivin, synovial sarcoma X breakpoint 2 (SSX2), TCR γ-substituted reading frame protein (TARP), TCR-β1 chain, TCR-β2 chain, TCR-δ chain, TCR-γ chain, TCRγ-δ, telomerase, TGFβR2, antigen recognized by TNT antibody, thyroid-stimulating hormone receptor (TSHR), Timol / HVCR1, tissue factor 1 (TF1), Tn ag, Tn antigen ((Tn) (Ag) or (GalNAca-Ser / Thr)), TNF receptor family member B cell maturation (BCMA), transglutaminase 5 (TGS5), transmembrane protease, TROP2, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7 associated (TEM7R), tumor protein p53 (p53), tumor-associated glycoprotein 72 (TAG72), tyrosinase, tyrosinase-associated protein 2 (TRP-2), urolytic protein 2 (UPK2), vascular endothelial growth factor receptor 2 (VEGFR2), V-myc avian myeloma virus oncogene neuroblastoma-derived homolog (MYCN), Wilms tumor protein (WT1) or X antigen family member 1A (XAGE1).

40. The viral particle of claim 39, wherein the targeting portion binds to CD7.

41. The viral particle of claim 40, wherein the targeting portion binding to CD7 comprises a polypeptide, the polypeptide comprising a sequence having at least 90% sequence identity with SEQ ID NO: 52, at least 95% sequence identity with SEQ ID NO: 52, at least 99% sequence identity with SEQ ID NO: 52, or a sequence as shown in SEQ ID NO:

52.

42. The viral particle according to any one of claims 34 to 41, wherein the targeting portion is attached to the viral surface by means of: IgG Fc stem; The envelope glycoprotein G or H of viruses of the Paramyxoviridae family, such as measlesviruses, such as measlesviruses, or Hennipaviruses, such as Nipah virus, cedar virus, or Hendra virus. Glycoproteins of viruses in the Rhabdoviridae family, such as vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), vesicular stomatitis virus (VSDV), parainfluenza virus, fall armyworm rhabdovirus isolate Sf G, Drosophila sigmavirus 10A, Wuhan insect virus 7, perch virus, or carp spring viremia virus; Glycoproteins of viruses in the Filoviridae family, such as Ebola virus; or Glycoproteins of viruses in the Arenaviridae family, such as Machupo virus.

43. A method for delivering a heterologous molecule to a target cell, the method comprising contacting the cell with a viral vector, the viral vector comprising: a) The variant VSV-G protein according to any one of claims 1 to 32; b) The targeting portion that binds to the target cells; and c) Nucleic acid molecules that encode the heterologous molecule.

44. A method of delivering a heterologous molecule to target cells in a subject, the method comprising administering a viral vector to the subject, the viral vector comprising: a) The variant VSV-G polypeptide according to any one of claims 1 to 32; b) The targeting portion that binds to the target cells; and c) Nucleic acid molecules that encode the heterologous molecule.

45. The method according to any one of claims 43 to 44, wherein the heterologous molecule is siRNA, shRNA, non-coding RNA, peptide, polypeptide, protein, viral payload, viral genome, chimeric antigen receptor ("CAR"), or a combination thereof.

46. ​​A method of treating cancer in a subject, the method comprising administering a viral vector to the subject, the viral vector comprising: a) The variant VSV-G polypeptide according to any one of claims 1 to 32; b) The targeting portion that binds to the target cells; and c) Nucleic acid molecules that encode the heterologous molecule.

47. The method of claim 46, wherein the heterologous molecule is a chimeric antigen receptor.

48. The method according to any one of claims 46 to 47, wherein the cancer is carcinoma, blastoma, sarcoma, leukemia, malignant lymphoma, benign tumor, malignant tumor, sarcoma, carcinoma, melanoma, breast cancer, prostate cancer, ovarian cancer, cervical cancer, skin cancer, pancreatic cancer, colorectal cancer, kidney cancer, liver cancer, brain cancer, lymphoma, leukemia, lung cancer, thyroid cancer, B-cell related cancer, or T-cell related cancer.