RU2020143425A

RU2020143425A - ANTIBODY AND DRUG CONJUGATES AS A SPLICING MODULATOR AND METHODS FOR THEIR APPLICATION

Info

Publication number: RU2020143425A
Application number: RU2020143425A
Authority: RU
Inventors: Эрмира ПАЦОЛЛИ; Сильвия БУОНАМИЧИ; Тхиванка Самаракун; Судип ПРАДЖАПАТИ; Натан Фишкин; Джеймс Паласино; Майкл Силер; Пин ЧЖУ; Эндрю Кук; Питер Смит; Сян Лю; Шелби Эллери; Доминик Рейнолдс; Лихуа ЮЙ; Чжэньхуа У; Шоуюн Пэн; Николас Каландра; Меган Шинан; Юнхун Сяо
Original assignee: Эйсай Ар Энд Ди Менеджмент Ко., Лтд.
Priority date: 2018-06-01
Filing date: 2019-05-31
Publication date: 2022-07-11

Claims

1. Antibody drug conjugate of formula (I):

Ab-(LD) p (I)

where

Ab is an antibody or antigen-binding fragment that targets a neoplastic cell;

D is a splicing modulator;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

2. The antibody-drug conjugate of claim 1, wherein the antibody or antigen-binding fragment specifically targets a neoplastic cell derived from a hematologic malignancy or solid tumor.

3. Antibody-drug conjugate according to claim 2,

wherein the hematologic malignancy is selected from B-cell malignancy, leukemia, lymphoma and myeloma, optionally the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma; and/or

where the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer and cancer esophagus.

4. Conjugate antibodies and drugs according to any one of paragraphs. 1-3, wherein the antibody or antigen-binding fragment targets a cell expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4 and/or STEAP1 .

5. Conjugate antibodies and drugs according to any one of paragraphs. 1-4, where

(a) the antibody or antigen-binding fragment targets a cell expressing HER2 and contains three complementarity-determining regions of the heavy chain (HCDR1, HCDR2 and HCDR3) containing the amino acid sequences under SEQ ID NO:1 (HCDR1), SEQ ID NO:2 ( HCDR2) and SEQ ID NO:3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) containing the amino acid sequences under SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2) and SEQ ID NO:6 (LCDR3);

(b) the antibody or antigen-binding fragment targets a cell expressing CD138 and contains three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) containing the amino acid sequences of SEQ ID NO:7 (HCDR1), SEQ ID NO:8 ( HCDR2) and SEQ ID NO:9 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) containing the amino acid sequences under SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2) and SEQ ID NO:12 (LCDR3); or

(c) the antibody or antigen-binding fragment targets a cell expressing EPHA2 and contains three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) containing the amino acid sequences of SEQ ID NO:13 (HCDR1), SEQ ID NO:14 ( HCDR2) and SEQ ID NO:15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) containing the amino acid sequences under SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2) and SEQ ID NO:18 (LCDR3).

6. The antibody-drug conjugate of claim 5, wherein

(a) the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:20;

(b) the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:22; or

(c) the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:24.

7. The antibody drug conjugate of claim 5 or 6, wherein the antibody or antigen binding fragment comprises a human IgG1 or IgG2a heavy chain constant region, and/or wherein the antibody or antigen binding fragment comprises a human Ig kappa light chain constant region.

8. Conjugate antibodies and drugs according to any one of paragraphs. 5-7, where

(a) the antibody or antigen-binding fragment contains a heavy chain containing the amino acid sequence of SEQ ID NO:25 and a light chain containing the amino acid sequence of SEQ ID NO:26;

(b) the antibody or antigen-binding fragment comprises a heavy chain containing the amino acid sequence of SEQ ID NO:27 and a light chain containing the amino acid sequence of SEQ ID NO:28; or

(c) the antibody or antigen-binding fragment contains a heavy chain containing the amino acid sequence of SEQ ID NO:29 and a light chain containing the amino acid sequence of SEQ ID NO:30.

9. Conjugate antibodies and drugs according to any one of paragraphs. 1-8, where the linker is a cleavable linker.

10. The antibody drug conjugate of claim 9, wherein the linker comprises a cleavable peptide fragment or a cleavable glucuronide fragment.

11. The antibody-drug conjugate of claim 10, wherein the cleavable peptide fragment is cleaved by the action of an enzyme.

12. Conjugate antibodies and drugs according to any one of paragraphs. 9-11, where the cleavable peptide fragment or linker provides an amino acid unit.

13. The antibody-drug conjugate of claim 12, wherein the amino acid unit comprises valine-citrulline (Val-Cit), valine-alanine (Val-Ala), glutamine-valine-citrulline (Glu-Val-Cit), or alanine-alanine -asparagine (Ala-Ala-Asn).

14. The antibody drug conjugate of claim 10 wherein the cleavable glucuronide moiety is cleaved by glucuronidase.

15. Conjugate antibodies and drugs according to any one of paragraphs. 1-14, where the linker provides at least one spacer unit.

16. The antibody drug conjugate of claim 15, wherein the spacer or linker comprises a polyethylene glycol (PEG) moiety or an alkyl moiety.

17. The antibody drug conjugate of claim 16 wherein the PEG fragment comprises -(PEG) m - and m is an integer from 1 to 10, optionally m is 2.

18. The antibody drug conjugate of claim 16, wherein the alkyl moiety comprises -(CH2) n - and n is an integer from 1 to 10, optionally n is 2, 5 or 6.

19. Conjugate antibodies and drugs according to any one of paragraphs. 15-18, where the spacer unit is attached to the antibody or antigen-binding fragment via a maleimide (Mal) fragment ("Mal spacer unit").

20. The antibody-drug conjugate of claim 19, wherein the Mal spacer unit is connected to the antibody or antigen-binding fragment via a cysteine residue on the antibody or antigen-binding fragment.

21. The antibody drug conjugate of claim 19 or 20, wherein the linker provides a Mal spacer unit and a cleavable peptide fragment, where the cleavable peptide fragment provides an amino acid unit.

22. An antibody-drug conjugate according to any one of paragraphs. 19-21, where the Mal-spacer unit provides maleimidocaproyl (MC).

23. Conjugate antibodies and drugs according to any one of paragraphs. 15-22, where the cleavable fragment in the linker is directly connected to the splicing modulator, or where the spacer unit attaches the cleavable fragment in the linker to the splicing modulator.

24. The antibody-drug conjugate of claim 23, wherein cleavage of the conjugate releases the splicing modulator from the antibody or antigen-binding fragment and the linker.

25. The antibody-drug conjugate of claim 23 or 24, wherein the spacer unit that attaches the cleavable fragment in the linker to the splicing modulator is self-cleaving.

26. Conjugate antibodies and drugs according to any one of paragraphs. 23-25, wherein the spacer linking the cleavable moiety in the linker to the splicing modulator is p-aminobenzyloxycarbonyl (pABC) or p-aminobenzyl (pAB).

27. The antibody-drug conjugate of claim 26, wherein the pABC or pAB attaches the cleavable fragment in the linker to the splicing modulator.

28. Conjugate antibodies and drugs according to any one of paragraphs. 1-8, where the linker is a non-cleavable linker.

29. The antibody drug conjugate of claim 28 wherein the linker is maleimidocaproyl (MC), MC-(PEG) ₂ , Mal-Hex, Mal-Et, or Mal-Et-O-Et.

30. The antibody-drug conjugate of claim 28 or 29, wherein the linker or Mal-spacer unit comprises maleimidocaproyl (MC) and at least one additional spacer unit, MC-(PEG) ₂ and at least one additional spacer unit, Mal-Et-O-Et and at least one additional spacer unit or Mal-Et-O-Et and at least one additional spacer unit.

31. Conjugate antibodies and drugs according to any one of paragraphs. 1-30, where the splicing modulator provides a splicing modulator of formula (II):

(II), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is absent or selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkylcarboxylic acid groups, C ₁ -C ₆ alkylhydroxy groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups, C ₃ -C ₈ heterocyclyl groups, -OC(=O)-(C ₁ -C ₆ alkyl) and -CD ₃ groups;

R ³ is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkylcarboxylic acid groups, C ₁ -C ₆ alkylhydroxy groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups, C ₃ -C ₈ heterocyclyl groups and -OC(=O)-(C ₁ -C ₆ alkyl) groups; and

each of R ⁴ , R ⁵ and R ⁸ is independently selected from hydrogen, hydroxyl groups, -O-(C ₁ -C ₆ alkyl) groups, -OC(=O)-(C ₁ -C ₆ alkyl) groups and C ₁ -C ₆ alkyl groups;

each of R ⁶ and R ⁷ is independently selected from hydrogen, -OR ¹⁷ , -OC(=O)-R ¹⁷ , -OC(=O)-NR ¹⁵ R ¹⁶ , C ₁ -C ₆ alkyl groups and -NR ¹⁵ R ¹⁶ ;

each of R ¹⁵ and R ¹⁶ is independently selected from hydrogen, R ¹⁷ , -C(=O)-R ¹⁷ and -C(=O)-OR ¹⁷ ;

R ¹⁷ is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups; and

Z is selected from

,

and

;

where each of R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁵ , R ¹⁶ and R ¹⁷ is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, C ₁ -C ₆ alkyl groups, -O-(C ₁ -C ₆ alkyl), -NR ¹⁵ R ¹⁶ , C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkylhydroxy groups, C ₁ -C ₆ alkylalkoxy groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups,

where at least one of R ⁶ and R ⁷ is hydrogen.

32. The antibody-drug conjugate of claim 31, wherein the linker is covalently attached to a splicing modulator ("L-D") and L-D have the structure of formula (II-A):

(II-A), or a pharmaceutically acceptable salt thereof,

where Z' is selected from

,

and

; and

where all other variables are as defined for formula (II).

33. An antibody-drug conjugate according to any one of paragraphs. 1-30, where the splicing modulator is a splicing modulator of formula (IV):

(IV), or a pharmaceutically acceptable salt thereof, wherein:

R ¹ is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkylcarboxylic acid groups, C ₁ -C ₆ alkylhydroxy groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups, C ₃ -C ₈ heterocyclyl groups, -OC(=O)-(C ₁ -C ₆ alkyl) and -CD ₃ groups;

each of R ¹⁵ and R ¹⁶ is independently selected from hydrogen, R ¹⁷ , -C(=O)-R ¹⁷ and -C(=O)-OR ¹⁷ ; and

R ¹⁷ is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups;

where at least one of R ⁶ and R ⁷ is hydrogen.

34. The antibody drug conjugate of claim 33, wherein the linker is covalently attached to a splicing modulator ("L-D") and L-D has the structure of formula (IV-A):

(IV-A), or a pharmaceutically acceptable salt thereof.

35. An antibody-drug conjugate according to any one of paragraphs. 1-30, where the splicing modulator is a splicing modulator of formula (VI):

(VI)

or a pharmaceutically acceptable salt thereof, where:

each of R1 and R9 is independently selected from hydrogen, C1-C6 alkyl groups, C ₁ -C ₆ alkyl alkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkyl carboxylic acid groups, C ₁ -C ₆ alkyl hydroxy groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups, C ₃ -C ₈ heterocyclyl groups, -OC(=O)-(C ₁ -C ₆ alkyl) and -CD ₃ groups;

R3 is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkyl alkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkyl carboxylic acid groups, C ₁ -C ₆ alkyl hydroxy groups, C ₃ -C ₈ cycloalkyl groups , benzyl groups, C ₃ -C ₈ heterocyclyl groups and -OC(=O)-(C ₁ -C ₆ alkyl) groups;

each of R4, R5 and R8 is independently selected from hydrogen, hydroxyl groups, -O-(C ₁ -C ₆ alkyl) groups, -OC(=O)-(C ₁ -C ₆ alkyl) groups and C ₁ -C ₆ alkyl groups;

each of R6 and R7 is independently selected from hydrogen, -O-R17, -OC(=O)-R17, -OC(=O)-NR15R16, C ₁ -C ₆ alkyl groups, -NR15R16 and a linker;

R10 is selected from hydrogen, C ₁ -C ₆ alkyl groups, -C(=O)-(C ₁ -C ₆ alkyl) and -CD ₃ groups;

each of R15 and R16 is independently selected from hydrogen, R17, -C(=O)-R17 and -C(=O)-O-R17;

R17 is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups; and

a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

where each of R1, R3, R4, R5, R6, R7, R8, R9, R10, R15, R16 and R17 is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, C ₁ -C ₆ alkyl groups, -O-(C ₁ -C ₆ alkyl), -NR15R16, C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkyl hydroxy groups, C ₁ -C ₆ alkyl alkoxy groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups;

where at least one of R6 and R7 is hydrogen; and

where both R1 and R9 cannot be absent.

36. The antibody drug conjugate of claim 35, wherein the linker is covalently attached to a splicing modulator ("L-D") and L-D have the structure of formula (VI-A):

(VI-A), or a pharmaceutically acceptable salt thereof.

37. An antibody-drug conjugate according to any one of paragraphs. 1-30, where the splicing modulator is a splicing modulator of formula (VIII):

(VIII), or a pharmaceutically acceptable salt thereof, wherein:

R ³ is selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkylcarboxylic acid groups, C ₁ -C ₆ alkylhydroxy groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups, C ₃ -C ₈ heterocyclyl groups and -OC(=O)-(C ₁ -C ₆ alkyl) groups;

R ⁴ is selected from hydrogen, hydroxyl groups, -O-(C ₁ -C ₆ alkyl), -OC(=O)-(C ₁ -C ₆ alkyl) groups, and C ₁ -C ₆ alkyl groups; and

R ¹⁰ is selected from 3-10-membered carbocycles and 3-10-membered heterocycles, each of which is substituted with 0-3 R ^a , where each R ^a is independently selected from halogen atoms, C ₁ -C ₆ alkyl groups, -O-( C ₁ -C ₆ )alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylhydroxy groups, -S(=O) _w -(4-7-membered heterocycles), 4-7-membered carbocycles and 4-7 -membered heterocycles;

where each of R ¹ , R ³ , R ⁴ , R ¹⁰ , R ¹⁵ , R ¹⁶ and R ¹⁷ is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, C ₁ -C ₆ alkyl groups, groups -O -(C ₁ -C ₆ alkyl), -NR ¹⁵ R ¹⁶ , C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkylhydroxy groups, C ₁ -C ₆ alkylalkoxy groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups; and

where each R ^a is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, -NR ¹⁵ R ¹⁶ , C ₁ -C ₆ alkyl groups, -(C=O)-(C ₁ -C ₆ alkyl) groups , groups -(C=O)-(C ₁ -C ₆ alkyl)-(C ₃ -C ₁₀ heterocyclyl), -S(=O) _w -(C ₃ -C ₈ heterocyclyl groups) and groups C ₁ -C ₆ alkylcarboxylic acids each substituted with 0, 1 or 2 groups independently selected from halogen atoms, hydroxyl groups, -NR ¹⁵ R ¹⁶ and C ₁ -C ₃ alkyl groups; and

w is 0, 1, or 2.

38. The antibody drug conjugate of claim 37, wherein the linker is covalently attached to a splicing modulator ("L-D") and L-D has the structure of formula (VIII-A):

(VIII-A), or a pharmaceutically acceptable salt thereof.

39. Conjugate antibodies and drugs according to any one of paragraphs. 1-30, where the splicing modulator provides D4.

40. The antibody drug conjugate of claim 39 wherein the splicing modulator comprises D4 and the linker comprises MC-Val-Cit-pABC or MC-β-glucuronide.

41. Conjugate antibodies and drugs according to any one of paragraphs. 1-30, where the splicing modulator provides D12.

42. The antibody drug conjugate of claim 41 wherein the splicing modulator comprises D12 and the linker comprises MC-Val-Cit-pABC or MC-β-glucuronide.

43. An antibody-drug conjugate according to any one of paragraphs. 1-30, where the splicing modulator provides for D15.

44. The antibody drug conjugate of claim 43, wherein the splicing modulator comprises D4 and the linker comprises MC-Val-Ala-pAB.

45. An antibody-drug conjugate according to any one of paragraphs. 1-30, wherein the splicing modulator comprises a pladienolide D or a pladienolide derivative comprising D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12, or D25.

46. The antibody drug conjugate of claim 45, wherein the pladienolide D or derivative provides for D2.

47. The antibody drug conjugate of claim 45 wherein the pladienolide D or derivative provides D1.

48. The antibody drug conjugate of claim 45, wherein the pladienolide D or derivative provides for D4.

49. The antibody drug conjugate of claim 45, wherein the pladienolide D or derivative provides for D12.

50. The antibody drug conjugate of claim 45, wherein the pladienolide D or derivative is a zwitterionic pladienolide D or a derivative containing D22 or D25.

51. An antibody-drug conjugate according to any one of paragraphs. 1-30, wherein the splicing modulator comprises pladienolide B or a pladienolide derivative comprising D9, D18, D19, or D13.

52. An antibody-drug conjugate according to any one of paragraphs. 1-30, wherein the splicing modulator comprises an aryl pladienolide comprising D15, D14, D16, D17, D26, or D33.

53. The antibody drug conjugate of claim 52, wherein the aryl pladienolide provides for D15.

54. The antibody-drug conjugate of claim 52, wherein the aryl pladienolide is a zwitterionic aryl pladienolide containing D33.

55. An antibody-drug conjugate according to any one of paragraphs. 1-30, where the splicing modulator provides D4, D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25, or D33.

56. An antibody-drug conjugate according to any one of paragraphs. 1-55 where p is from 1 to 10.

57. An antibody-drug conjugate according to any one of paragraphs. 1-56 where p is 4 or 8.

58. Antibody drug conjugate of formula (I):

Ab-(LD) p (I),

where

Ab is an antibody or antigen-binding fragment that targets a neoplastic cell;

D is D2;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

59. Antibody drug conjugate of formula (I):

Ab-(LD) p I),

where

Ab is an antibody or antigen-binding fragment that targets a neoplastic cell;

D is D1;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

60. Antibody drug conjugate of formula (I):

Ab-(LD) p (I),

where

Ab is an antibody or antigen-binding fragment that targets a neoplastic cell;

D is D4;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

61. Antibody drug conjugate of formula (I):

Ab-(LD) p (I),

where

Ab is an antibody or antigen-binding fragment that targets a neoplastic cell;

D is D12;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

62. Antibody-drug conjugate of formula (I):

Ab-(LD) p (I),

where

Ab is an antibody or antigen-binding fragment that targets a neoplastic cell;

D is D15;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

63. An antibody-drug conjugate having the structure

(Ab-ADL1-D1),

where Ab is an antibody or antigennegative fragment covalently linked to the maleimide group of ADL1-D1 through the sulfur atom of the thiol group on the antibody or antigennegative fragment.

64. An antibody-drug conjugate having the structure

(Ab-ADL1-D4),

where Ab is an antibody or antigennegative fragment covalently linked to the maleimide group of ADL1-D4 through the sulfur atom of the thiol group on the antibody or antigennegative fragment.

65. An antibody-drug conjugate having the structure

(Ab-ADL1-D12),

where Ab is an antibody or antigennegative fragment covalently linked to the maleimide group of ADL1-D12 through the sulfur atom of the thiol group on the antibody or antigennegative fragment.

66. An antibody-drug conjugate having the structure

(Ab-ADL5-D2),

where Ab is an antibody or antigennegative fragment covalently linked to the maleimide group of ADL5-D2 through the sulfur atom of the thiol group on the antibody or antigennegative fragment.

67. An antibody-drug conjugate having the structure

(Ab-ADL5-D15),

where Ab is an antibody or antigennegative fragment covalently linked to the maleimide group of ADL5-D15 through a thiol sulfur atom on the antibody or antigennegative fragment.

68. An antibody-drug conjugate having the structure

(Ab-ADL13-D4),

where Ab is an antibody or antigen-binding fragment covalently linked to the maleimide group of ADL13-D4 through a thiol sulfur atom on the antibody or antigen-binding fragment.

69. Conjugate antibodies and drugs according to any one of paragraphs. 58-68, wherein the antibody or antigen-binding fragment targets a cell expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4 and/or STEAP1 .

70. An antibody-drug conjugate according to any one of paragraphs. 58-69, where

71. An antibody-drug conjugate according to claim 70, where

72. The antibody drug conjugate of claim 70 or 71, wherein the antibody or antigen-binding fragment comprises a human IgG1 or IgG2a heavy chain constant region, and/or wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

73. An antibody-drug conjugate according to any one of paragraphs. 70-72, where

74. Antibody drug conjugate of formula (I):

Ab-(LD) p (I),

where

Ab is an anti-HER2 antibody or antigen binding fragment containing three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) containing the amino acid sequences under SEQ ID NO:1 (HCDR1), SEQ ID NO:2 (HCDR2) and SEQ ID NO :3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) containing the amino acid sequences under SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2) and SEQ ID NO:6 (LCDR3);

D is D2, D1, D4, D12 or D15;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

75. The antibody-drug conjugate of claim 74, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:20.

76. Antibody-drug conjugate of formula (I):

Ab-(LD) _p (I),

where

Ab is an anti-CD138 antibody or antigen binding fragment containing three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) containing the amino acid sequences under SEQ ID NO:7 (HCDR1), SEQ ID NO:8 (HCDR2) and SEQ ID NO :9 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) containing the amino acid sequences under SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2) and SEQ ID NO:12 (LCDR3);

D is D2, D1, D4, D12 or D15;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

77. The antibody-drug conjugate of claim 76, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:22.

78. Antibody-drug conjugate of formula (I):

Ab-(LD) _p (I),

where

Ab is an anti-EPHA2 antibody or antigen binding fragment containing three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) containing the amino acid sequences under SEQ ID NO:13 (HCDR1), SEQ ID NO:14 (HCDR2) and SEQ ID NO :15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) containing the amino acid sequences under SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2) and SEQ ID NO:18 (LCDR3);

D is D2, D1, D4, D12 or D15;

L is a linker that covalently attaches Ab to D; and

p is an integer from 1 to 15.

79. The antibody-drug conjugate of claim 78, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:24.

80. Conjugate antibodies and drugs according to any one of paragraphs. 58-62 or 69-79, where the linker is a cleavable linker.

81. The antibody-drug conjugate of claim 80 wherein the cleavable linker comprises MC-Val-Cit-pABC, MC-Val-Ala-pABC, MC-Val-Ala-pAB, MC-Glu-Val-Cit-pABC, or MC-Ala-Ala-Asn-pABC.

82. An antibody-drug conjugate according to any one of paragraphs. 58-62 or 69-79, where the linker is a non-cleavable linker.

83. The antibody-drug conjugate of claim 82, wherein the non-cleavable linker comprises Mal-Hex, Mal-Et, or Mal-Et-O-Et.

84. An antibody-drug conjugate according to any one of paragraphs. 58-83, where p is from 1 to 10.

85. An antibody-drug conjugate according to any one of paragraphs. 58-84 where p is 4 or 8.

86. A composition containing multiple copies of the antibody-drug conjugate according to any one of paragraphs. 1-85, where the average p value of the antibody-drug conjugates in the composition is from about 3.5 to about 5.5.

87. The composition of claim 86, wherein the average p value of the antibody drug conjugates in the composition is approximately 4.

88. Composition containing multiple copies of the conjugate of antibodies and drugs according to any one of paragraphs. 1-85, where the average p value of the antibody-drug conjugates in the composition is from about 7 to about 9.

89. The composition of claim 88, wherein the average p value of the antibody drug conjugates in the composition is approximately 8.

90. A method of treating a subject who has or is suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89.

91. The method of claim 90 wherein the neoplastic disorder is a hematologic malignancy or a solid tumor.

92. The method according to p. 91,

wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma, optionally the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma; and/or

93. The method according to any one of paragraphs. 90-92, wherein the subject has one or more neoplastic cells that express the target antigen, wherein the target antigen is HER2, CD138, or EPHA2.

94. The method according to p. 93,

where one or more neoplastic cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma or salivary duct carcinomas expressing HER2, optionally the lung cancer being lung adenocarcinoma and/or the uterine cancer being endometrial serous carcinoma of the uterus;

where one or more neoplastic cells are derived from multiple myeloma expressing CD138; or

wherein one or more neoplastic cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

95. A method of reducing or suppressing tumor growth in a subject who has a neoplastic disorder or suspected of having it, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89.

96. The method of claim 95, wherein the tumor contains one or more neoplastic cells that express the target antigen, wherein the target antigen is HER2, CD138, or EPHA2.

97. The method of claim 96, wherein one or more neoplastic cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma or salivary duct carcinomas expressing HER2, optionally the lung cancer being lung adenocarcinoma and/or the uterine cancer being endometrial serous carcinoma of the uterus;

98. The use of an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89 in the treatment of a neoplastic disorder, wherein the neoplastic disorder is characterized by having one or more neoplastic cells that express a target antigen, wherein the target antigen is HER2, CD138, or EPHA2.

99. Application according to paragraph 98,

where the neoplastic disorder is breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma or salivary duct carcinoma expressing HER2, optionally the lung cancer being lung adenocarcinoma and/or the uterine cancer being endometrial serous carcinoma of the uterus;

where the neoplastic disorder is multiple myeloma expressing CD138; or

where the neoplastic disorder is breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer or esophageal cancer expressing EPHA2.

100. The use of an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89 in a method of making a medicament for the treatment of a neoplastic disorder, wherein the neoplastic disorder is characterized by having one or more neoplastic cells that express a target antigen, wherein the target antigen is HER2, CD138, or EPHA2.

101. Use according to claim 100, where the neoplastic disorder is breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma or salivary duct carcinoma expressing HER2, optionally the lung cancer being lung adenocarcinoma and/or the uterine cancer being endometrial serous carcinoma of the uterus;

where the neoplastic disorder is multiple myeloma expressing CD138; or

102. A pharmaceutical composition containing an antibody-drug conjugate according to any one of paragraphs. 1-85 or a composition according to any one of paragraphs. 86-89 and a pharmaceutically acceptable carrier.

103. A method for producing an antibody-drug conjugate according to any one of paragraphs. 1-85, comprising reacting the antibody or antigen-binding fragment with a linker attached to the splicing modulator under conditions that allow conjugation.

104. Compound of formula (III):

(III), or a pharmaceutically acceptable salt thereof, wherein:

R ² is absent or is a linker;

each of R ⁶ and R ⁷ is independently selected from hydrogen, -OR ¹⁷ , -OC(=O)-R ¹⁷ , -OC(=O)-NR ¹⁵ R ¹⁶ , C ₁ -C ₆ alkyl groups, -NR ¹⁵ R ¹⁶ and linker;

Z'' selected from

,

and

;

where each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁵ , R ¹⁶ and R ¹⁷ is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups , C ₁ -C ₆ alkyl groups, -O-(C ₁ -C ₆ alkyl), -NR ¹⁵ R ¹⁶ , C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkylhydroxy groups, C ₁ -C ₆ alkylalkoxy groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups;

where at least one of R ⁶ and R ⁷ is hydrogen; and

where, if R ² is a linker, then neither R ⁶ nor R ⁷ is a linker, and if R ⁶ or R ⁷ is a linker, then R ² is absent.

105. Compound of formula (V):

(V), or a pharmaceutically acceptable salt thereof, wherein:

R ² is absent or is a linker;

where each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ¹⁵ , R ¹⁶ , and R ¹⁷ is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, C ₁ -C ₆ alkyl groups, groups -O-(C ₁ -C ₆ alkyl), -NR ¹⁵ R ¹⁶ , C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkylhydroxy groups, C ₁ -C ₆ alkylalkoxy groups , benzyl groups and C ₃ -C ₈ heterocyclyl groups;

where at least one of R ⁶ and R ⁷ is hydrogen; and

106. Compound of formula (VII):

(VII), or a pharmaceutically acceptable salt thereof, wherein:

each of R ¹ and R ⁹ is independently absent or selected from hydrogen, C ₁ -C ₆ alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylamino groups, C ₁ -C ₆ alkylcarboxylic acid groups, C ₁ -C ₆ alkylhydroxy groups, C ₃ -C ₈ cycloalkyl groups, benzyl groups, C ₃ -C ₈ heterocyclyl groups, -OC(=O)-(C ₁ -C ₆ alkyl) and -CD ₃ groups;

R ² is absent or is a linker;

R ¹⁰ is selected from hydrogen, C ₁ -C ₆ alkyl groups, -C(=O)-(C ₁ -C ₆ alkyl) and -CD ₃ groups;

a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

where each of R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹⁵ , R ¹⁶ , and R ¹⁷ is independently substituted with 0-3 groups, independently selected from halogen atoms, hydroxyl groups, C ₁ -C ₆ alkyl groups, -O-(C ₁ -C ₆ alkyl), -NR ¹⁵ R ¹⁶ groups, C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkyl hydroxy groups, C ₁ -C ₆ alkylalkoxy groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups;

where at least one of R ⁶ and R ⁷ is hydrogen;

where, if R ² is a linker, then neither R ⁶ nor R ⁷ is a linker, and if R ⁶ or R ⁷ is a linker, then R ² is absent; and

where both R ¹ and R ⁹ cannot be absent.

107. Compound of formula (IX):

(IX), or a pharmaceutically acceptable salt thereof, wherein:

R ² is a linker;

R ⁴ is selected from hydrogen, hydroxyl groups, -O-(C ₁ -C ₆ alkyl), -OC(=O)-(C ₁ -C ₆ alkyl) groups, and C ₁ -C ₆ alkyl groups;

R ¹⁰ is selected from 3-10 membered carbocycles and 3-10 membered heterocycles, each of which is substituted with 0-3 R ^a , where each R ^a is independently selected from halogen atoms, C ₁ -C ₆ alkyl groups, -O-( C ₁ -C ₆ )alkyl groups, C ₁ -C ₆ alkylalkoxy groups, C ₁ -C ₆ alkylhydroxy groups, -S(=O) _w -(4-7-membered heterocycles), 4-7-membered carbocycles and 4-7 -membered heterocycles;

where each of R ¹ , R ² , R ³ , R ⁴ , R ¹⁰ , R ¹⁵ , R ¹⁶ and R ¹⁷ is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, C ₁ -C ₆ alkyl groups, -O-(C ₁ -C ₆ alkyl), -NR ¹⁵ R ¹⁶ , C ₃ -C ₈ cycloalkyl groups, C ₁ -C ₆ alkylhydroxy groups, C ₁ -C ₆ alkylalkoxy groups, benzyl groups and C ₃ -C ₈ heterocyclyl groups groups; and

where each R ^a is independently substituted with 0-3 groups independently selected from halogen atoms, hydroxyl groups, -NR ¹⁵ R ¹⁶ , C ₁ -C ₆ alkyl groups, -(C=O)-(C ₁ -C ₆ alkyl) groups , groups -(C=O)-(C ₁ -C ₆ alkyl)-(C ₃ -C ₁₀ heterocyclyl), -S(=O) _w -(C ₃ -C ₈ heterocyclyl groups and C ₁ -C ₆ groups alkylcarboxylic acid each substituted with 0, 1 or 2 groups independently selected from halogen atoms, hydroxyl groups, -NR ¹⁵ R ¹⁶ and C ₁ -C ₃ alkyl groups.

108. Compound selected from:

(a) compounds of formula

(D4) and its pharmaceutically acceptable salts;

(b) compounds of the formula

(D8) and its pharmaceutically acceptable salts;

(c) compounds of the formula

(D9) and its pharmaceutically acceptable salts;

(d) compounds of the formula

(D10) and its pharmaceutically acceptable salts;

(e) compounds of the formula

(D13) and its pharmaceutically acceptable salts;

(f) compounds of formula

(D18) and its pharmaceutically acceptable salts;

(g) compounds of the formula

(D19) and its pharmaceutically acceptable salts;

(h) compounds of the formula

(D20) and its pharmaceutically acceptable salts;

(i) compounds of the formula

(D21) and its pharmaceutically acceptable salts;

(j) compounds of the formula

(D22) and its pharmaceutically acceptable salts;

(k) compounds of formula

(D25) and its pharmaceutically acceptable salts;

(l) compounds of the formula

(D33) and its pharmaceutically acceptable salts.

109. A pharmaceutical composition containing a compound according to claim 108 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

110. A method of treating a subject who has or is suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of a compound of claim 108 or a pharmaceutical composition of claim 109.

111. A method of reducing or inhibiting tumor growth in a subject who has or is suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of a compound of claim 108 or a pharmaceutical composition of claim 109.

112. A method of inducing the formation of at least one neoantigen, including bringing the neoplastic cell into contact with an effective amount of a splicing modulator, an antibody-drug conjugate according to any one of paragraphs. 1-85 or a composition according to any one of paragraphs. 86-89, whereby the production of at least one neoantigen is induced.

113. The method according to p. 112, where at least one neoantigen contains the amino acid sequence under any of SEQ ID NO: 37-65.

114. The method of claim 112 or 113, wherein the neoplastic cell is derived from a hematologic malignancy or solid tumor.

115. The method according to item 114,

116. a method of inducing the formation of at least one neoantigen and / or the development of a T-cell response in a subject who has a neoplastic disorder or a suspicion of its presence, including administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89.

117. A method of treating a subject who has a neoplastic disorder or suspected of having it, comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89, wherein administration of the splicing modulator, antibody-drug conjugate or composition induces the formation of at least one neoantigen and/or the development of a T cell response.

118. The method according to p. 116 or 117, where at least one neoantigen contains the amino acid sequence under any of SEQ ID NO: 37-65.

119. The method according to any one of paragraphs. 116-118, further comprising administering at least one additional therapy.

120. The method according to any one of paragraphs. 116-119, wherein the amount of splicing modulator, antibody drug conjugate, composition, and/or at least one additional therapeutic agent administered is reduced due to induction of at least one neoantigen and/or development of a T-cell response compared to the standard dosage of the modulator splicing, antibody-drug conjugate, composition and/or at least one additional therapy.

121. The method according to any one of paragraphs. 116-120, where the amount of splicing modulator, antibody-drug conjugate, composition and/or at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% of the standard dosage of the splicing modulator, antibody drug conjugate, composition, and/or at least one additional therapy.

122. The method according to any one of paragraphs. 116-121, where the frequency of administration of the splicing modulator, antibody-drug conjugate, composition and/or at least one additional therapy is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard schedule for administering the splicing modulator, antibody-drug conjugate, composition, and/or at least one additional therapy.

123. The method according to any one of paragraphs. 116-122, wherein the administered amount and/or dosage of the splicing modulator, antibody drug conjugate, composition, and/or at least one additional therapy result in lower systemic toxicity and/or improved tolerability.

124. The method according to any one of paragraphs. 119-123, wherein administration of the splicing modulator, antibody-drug conjugate or composition is initiated prior to administration, after administration, or concurrently with administration of at least one additional therapy.

125. The method according to any one of paragraphs. 116-124, wherein administration of the splicing modulator, antibody drug conjugate, composition, and/or at least one additional therapy is repeated at least once after the initial administration.

126. The method of claim 125, wherein the amount of splicing modulator, antibody drug conjugate, composition, and/or at least one additional therapy used for repeated administration is reduced compared to the amount used for initial administration.

127. The method according to any one of paragraphs. 119-126, wherein at least one additional therapy comprises administering a checkpoint inhibitor.

128. The method according to p. 127, where the subject is characterized by intolerance, resistance or insufficient susceptibility to the checkpoint inhibitor when administered alone.

129. The method of claim 127 or 128, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3, and/or KIR, and/or where the checkpoint inhibitor provides for an inhibitor of the antigen pathway 4, associated with cytotoxic T-lymphocytes (CTLA4), an inhibitor of the programmed cell death protein 1 (PD1) pathway, or a CTLA4 inhibitor and a PD1 inhibitor.

130. The method according to p. 129,

where the CTLA4 inhibitor is an antibody to CTLA4, optionally the antibody to CTLA4 is ipilimumab;

where the PD1 inhibitor is an antibody to PD1, optionally the antibody to PD1 is nivolumab; and/or

where the PD1 inhibitor is an anti-PDL1 antibody, optionally the anti-PDL1 antibody is atezolizumab.

131. The method according to any one of paragraphs. 119-126, where at least one additional therapy involves the introduction of a neoantigenic vaccine.

132. The method of claim 131, wherein the neoantigenic vaccine comprises at least one neoantigenic peptide or neoantigenic mRNA.

133. The method of claim 132, wherein the length of at least one neoantigenic peptide is in the range of about 10 to about 50, or about 10 to about 35 amino acids.

134. The method of claim 132 or 133, wherein the at least one neoantigenic peptide is in the range of about 15 to about 25 amino acids in length.

135. The method according to any one of paragraphs. 132-134, where at least one neoantigenic peptide or neoantigenic mRNA contains or encodes one or more than one neoantigenic sequence.

136. The method of claim 135, wherein the neoantigenic sequence is a subject-specific neoantigenic sequence.

137. The method of claim 135, wherein the neoantigenic sequence is a universal neoantigenic sequence.

138. The method according to any one of paragraphs. 135-137, where the neoantigenic sequence contains the amino acid sequence under any of SEQ ID NOs: 37-65.

139. The method according to any one of paragraphs. 135-137, where the neoantigenic sequence contains the amino acid sequence under any of SEQ ID NOs: 66-93 or SEQ ID NOs: 74-77, or the antigenic portion under any of SEQ ID NOs: 66-93 or SEQ ID NOs: 74-77 .

140. The method of claim 138 or 139, wherein the neoantigenic sequence and/or antigenic portion has been identified by sequencing at least one neoantigen that has been induced in the subject by administering an effective amount of a splicing modulator, antibody drug conjugate or composition.

141. The method of claim 140, wherein the neoantigenic sequence and/or antigenic portion is capable of binding to at least one HLA allele expressed in the subject.

142. The method of claim 138 or 139, wherein the neoantigenic sequence and/or antigenic portion is a universal neoantigenic vaccine.

143. The method according to p. 142, where the neoantigenic sequence and / or antigenic part is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from a neoplastic disorder.

144. The method of claim 142 or 143, wherein the neoantigenic sequence and/or antigenic portion is capable of eliciting a T-cell response to a tumor present in at least 1%, at least 5%, or at least 10% of a population of affected subjects. neoplastic disorder.

145. The method according to any one of paragraphs. 142-144, wherein at least one neoantigenic peptide or neoantigenic mRNA contains a neoantigenic sequence whose formation is induced by contacting a neoplastic cell with an effective amount of a splicing modulator, antibody drug conjugate or composition.

146. The method according to any one of paragraphs. 131-145, wherein the neoantigenic vaccine comprises at least one neoantigenic peptide or neoantigenic mRNA and a pharmaceutically acceptable carrier.

147. The method of claim 146 wherein at least one neoantigenic peptide or neoantigenic mRNA is linked to a pharmaceutically acceptable carrier.

148. The method of claim 146 or 147, wherein the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, fisurella hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, toxoid, or attenuated toxoid derivative, cytokine, and chemokine.

149. The method according to any one of paragraphs. 146-148, wherein the neoantigenic peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

150. The method according to any one of paragraphs. 146-148, wherein the neoantigenic peptide and the pharmaceutically acceptable carrier are expressed as a fusion protein.

151. The method according to any one of paragraphs. 131-145, where the neoantigenic vaccine contains at least one neoantigenic peptide or neoantigenic mRNA and a pharmaceutically acceptable diluent.

152. The method according to any one of paragraphs. 131-145, where the neoantigenic vaccine contains at least one neoantigenic peptide or neoantigenic mRNA and a pharmaceutically acceptable adjuvant.

153. The method according to any one of paragraphs. 131-152, where the neoantigenic mRNA is encapsulated with an encapsulating agent.

154. The method of claim 153 wherein the encapsulating agent is a liposome or nanoparticle.

155. The method according to any one of paragraphs. 119-126, wherein at least one additional therapeutic agent involves the administration of a cytokine or cytokine analog.

156. The method according to p. 155, where the subject is characterized by intolerance, resistance or insufficient susceptibility to a cytokine or cytokine analogue when administered separately.

157. The method of claim 155 or 156, wherein the cytokine or cytokine analog provides an enhancer for T cells.

158. The method according to any one of paragraphs. 155-157, wherein the cytokine or cytokine analog is IL-2, IL-10, IL-12, IL-15, IFNγ and/or TNFα.

159. The method according to any one of paragraphs. 119-126, wherein at least one additional therapy involves the administration of engineered T cells that target the tumor.

160. The method according to any one of paragraphs. 116-159, further comprising detecting one or more neoantigens and/or a T cell response in the subject after administration of the splicing modulator, antibody drug conjugate or composition.

161. The method of claim 160, further comprising continuing to administer the splicing modulator, antibody drug conjugate or composition if one or more neoantigens and/or a T cell response is detected.

162. The method of claim 160 or 161, further comprising continuing to administer the splicing modulator, antibody-drug conjugate, or composition at a reduced frequency and/or at a reduced dosage if one or more neoantigens and/or a T cell response is detected.

163. The method according to any one of paragraphs. 160-162, wherein the detection of one or more neoantigens and/or a T cell response in a subject is indicative of efficacy of treatment with a splicing modulator, antibody drug conjugate or composition.

164. The method according to any one of paragraphs. 116-163, wherein the subject has a non-synonymous mutation load of approximately 150 mutations or less.

165. The method according to any one of paragraphs. 116-164, where the neoplastic disorder is a hematological malignancy or a solid tumor.

166. The method according to item 165,

167. A method of treating a subject who has or is suspected of having a neoplastic disorder, comprising:

(a) administering to the subject an effective amount of the splicing modulator, antibody-drug conjugate of any one of claims. 1-85 or compositions according to any one of paragraphs. 86-89, wherein administration of the splicing modulator, antibody-drug conjugate or composition induces the formation of at least one neoantigen and/or the development of a T cell response;

(b) detecting one or more neoantigens and/or a T cell response in the subject following administration of the splicing modulator, antibody drug conjugate or composition; and

(c) continuing to administer the splicing modulator, antibody drug conjugate or composition if one or more neoantigens and/or a T cell response is detected.

168. The method of claim 167, wherein the detection of one or more neoantigens and/or a T cell response in the subject indicates efficacy of treatment with the splicing modulator, antibody drug conjugate or composition.

169. The method according to p. 167 or 168, where one or more neoantigens contain the amino acid sequence under any of SEQ ID NO: 37-65.

170. A method of treating a subject who has or is suspected of having a neoplastic disorder, comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89 and at least one additional therapy.

171. The method of claim 170, wherein the at least one additional therapy comprises at least one, at least two, at least three, at least four, or at least five additional therapies.

172. The method of claim 170 or 171, wherein administration of the splicing modulator, antibody-drug conjugate or composition induces the formation of at least one neoantigen and/or the development of a T cell response.

173. A method of treating a subject who has a neoplastic disorder or suspicion of its presence, including:

(a) administering to the subject an effective amount of the splicing modulator, antibody-drug conjugate of any one of claims. 1-85 or compositions according to any one of paragraphs. 86-89;

(b) detection of one or more neoantigens in the subject after administration of the splicing modulator, antibody-drug conjugate or composition;

(c) comparing one or more neoantigens to a panel of universal neoantigens; and

(d) administering to the subject a universal neoantigen vaccine comprising at least one universal neoantigen present in the subject.

174. The method of claim 173, wherein the universal neoantigen vaccine is administered alone or in combination with at least one additional therapy.

175. The method of claim 174, wherein the at least one additional therapy comprises at least one, at least two, at least three, at least four, or at least five additional therapies.

176. The method of claim 174 or 175, wherein at least one additional therapeutic agent comprises administering a checkpoint inhibitor.

177. A method for identifying at least one neoantigen, including:

(a) bringing the neoplastic cell into contact with an effective amount of splicing modulator, antibody-drug conjugate according to any one of paragraphs. 1-85 or compositions according to any one of paragraphs. 86-89;

(b) detecting at least one alternatively spliced mRNA transcript after contacting the neoplastic cell with a splicing modulator, antibody drug conjugate or composition;

(c) predicting the translation of at least one alternatively spliced mRNA transcript into at least one peptide; and

(d) comparing at least one peptide to a reference proteome, where at least one neoantigen is identified if at least one peptide does not match any peptides in the reference proteome.

178. A method for producing a neoantigenic vaccine, including:

(a) identifying at least one neoantigen using the method of claim 177; and

(b) formulating at least one neoantigen together with a pharmaceutically acceptable carrier, diluent or adjuvant.

179. The method according to p. 178, where at least one neoantigen contains the amino acid sequence under any of SEQ ID NO: 37-65.

180. The method according to p. 178, where at least one neoantigen contains an amino acid sequence under any of SEQ ID NO: 66-93 or SEQ ID NO: 74-77, or an antigenic part under any of SEQ ID NO: 66-93 or SEQ ID NO: 74-77.

181. The method according to any one of paragraphs. 178-180, wherein the length of at least one neoantigen and/or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

182. The method according to any one of paragraphs. 178-181, wherein at least one neoantigen and/or antigenic portion is associated with a pharmaceutically acceptable carrier.

183. The method according to any one of paragraphs. 178-182, wherein the pharmaceutically acceptable carrier is selected from peptide, serum albumin, fisurella hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, toxoid or attenuated toxoid derivative, cytokine, and chemokine.

184. Neoantigenic vaccine containing at least one neoantigenic peptide, where at least one neoantigenic peptide contains a modified or new neoantigenic sequence, the formation of which is induced by bringing a neoplastic cell into contact with an effective amount of a splicing modulator, an antibody-drug conjugate according to any one of paragraphs . 1-85 or compositions according to any one of paragraphs. 86-89.

185. Neoantigenic vaccine containing at least one neoantigenic mRNA, where at least one neoantigenic mRNA encodes a modified or new neoantigenic sequence, the formation of which is induced by bringing a neoplastic cell into contact with an effective amount of a splicing modulator, an antibody-drug conjugate according to any one of paragraphs . 1-85 or compositions according to any one of paragraphs. 86-89.