Antibodies and conjugates against prostaglandin F2 receptor inhibitors and their use

Abstract

This disclosure provides an antibody and a method for preparing and using the same, wherein the antibody binds to a PTGFRN on a cell.

Claims

1. a) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 1, 2, and 3, and light chain variable regions including CDR sequences, sequence numbers 7, 8, and 9, respectively; b) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 4, 5, and 6, and light chain variable regions including CDR sequences, sequence numbers 7, 8, and 9, respectively; c) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 4, 5, and 6, and light chain variable regions including CDR sequences, sequence numbers 7, 8, and 9, respectively; d) The heavy chain variable region of Sequence ID No. 189 and the light chain variable region of Sequence ID No. 190; e) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 18, 19, and 20, and light chain variable regions including CDR sequences, sequence numbers 24, 25, and 26, respectively; f) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 21, 22, and 23, and light chain variable regions including CDR sequences, sequence numbers 24, 25, and 26, respectively; g) Each of the following: a heavy chain variable region including a complementarity-determining region (CDR) sequence, sequence numbers 27, 28, and 29 or 30, and a light chain variable region including a CDR sequence, sequence numbers 31 or 32, and 33, and 34 or 35; h) The heavy chain variable region of Sequence ID No. 191 and the light chain variable region of Sequence ID No. 192; i) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 36, 37, and 38, and light chain variable regions including CDR sequences, sequence numbers 42, 43, and 44, respectively; j) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 39, 40, and 41, and light chain variable regions including CDR sequences, sequence numbers 42, 43, and 44, respectively; k) A heavy chain variable region comprising a complementarity-determining region (CDR) sequence, sequence numbers 45, and 46 or 47, and 48 or 49, respectively, and a light chain variable region comprising a CDR sequence, sequence numbers 50, and 51, and 52 or 53; l) The heavy chain variable region of sequence number 193 and 194; m) heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 54, 55, and 56, and light chain variable regions including CDR sequences, sequence numbers 60, 61, and 62, respectively; n) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 57, 58, and 59, and light chain variable regions including CDR sequences, sequence numbers 60, 61, and 62, respectively; o) Heavy chain variable regions including complementarity-determining region (CDR) sequences, sequence numbers 63, 64 or 65, and 66, respectively, and light chain variable regions including CDR sequences, sequence numbers 67, 68, and 69 or 70; p) The heavy chain variable region of Sequence ID No. 195 and the light chain variable region of Sequence ID No. 196; or one of the derivatives a) to p) An isolated antibody or antigen-binding fragment thereof comprising, wherein the derivative may contain 1 to 4 amino acid substitutions in at least one CDR, preferably the substitutions being a conserved amino acid sequence. The antibody or its derivative specifically binds to a human prostaglandin F2 receptor inhibitor (PTGFRN). An antibody or its antigen-binding fragment.

2. The antibody according to claim 1, which binds to cells expressing PTGFRN in vitro and / or in vivo.

3. An antibody that competes with the antibody described in claim 1 for binding to PTGFRN on cells.

4. A combination of antibodies according to any one of claims 1 to 3.

5. The antibody according to any one of claims 1 to 4, which is an isolated monoclonal antibody.

6. The antibody according to claim 5, wherein the monoclonal antibody is a human monoclonal antibody.

7. An antibody according to any one of claims 1 to 6, derived from human antibodies, human IgG, human IgG1, human IgG2, human IgG2a, human IgG2b, human IgG3, human IgG4, human IgM, human IgA, human IgA1, human IgA2, human IgD, human IgE, canine antibodies, canine IgGA, canine IgGGB, canine IgGC, canine IgGD, chicken antibodies, chicken IgA, chicken IgD, chicken IgE, chicken IgG, chicken IgM, chicken IgY, cat antibodies, goat antibodies, goat IgG, mouse antibodies, mouse IgG, pig antibodies, rat antibodies, llama antibodies, alpaca antibodies, shark antibodies, and camel antibodies.

8. A derivative of the antibody according to any one of claims 1 to 7, wherein F ab F ab2 , Fab' single chain antibody, F v Derivatives of antibodies, which may be selected from the group consisting of single-chain antibodies, monospecific antibodies, bispecific antibodies, trispecific antibodies, multispecific antibodies, polyvalent antibodies, chimeric antibodies, canine-human chimeric antibodies, canine-mouse chimeric antibodies, antibodies containing canine Fc, humanized antibodies, human antibodies, canine antibodies, CDR-grafted antibodies, shark antibodies, and nanobodies.

9. A derivative of an antibody according to any one of claims 1 to 8, comprising a detectable label immobilely bound thereto, wherein the detectable label is fluorescein, DyLight, Cy3, Cy5, FITC, HiLyte Fluor 555, HiLyte Fluor 647, 5-carboxy-2,7-dichlorofluorescein, 5-carboxyfluorescein, 5-FAM, hydroxytryptamine, 5-hydroxytryptamine (5-HAT), 6-carboxyfluorescein (6-FAM), FITC, 6-carboxy-1,4-dichloro-2',7'-dichlorofluorescein (TET), 6-carboxy-1,4-dichloro-2',4',5',7'-tetrachlorofluorescein (HEX), 6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein (6-JOE), Alexa fluor, Alexa fluor 350, Alexa fluor 405, Alexa fluor 430, Alexa fluor 488, Alexa fluor 500, Alexa fluor 514, Alexa fluor 532, Alexa fluor 546, Alexa fluor 555, Alexa fluor 568, Alexa fluor 594, Alexa fluor 610, Alexa fluor 633, Alexa fluor 635, Alexa fluor 647, Alexa fluor 660, Alexa fluor 680, Alexa fluor 700, Alexa fluor 750, BODIPY fluorophore, BODIPY 492 / 515, BODIPY 493 / 503, BODIPY 500 / 510, BODIPY 505 / 515, BODIPY 530 / 550, BODIPY 542 / 563, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650-X, BODIPY 650 / 665-X, BODIPY 665 / 676, FL, FL ATP, FI-Ceramide, R6G SE, TMR, TMR-X Conjugate, TMR-X, SE, TR, TR ATP, TR-X SE, Rhodamine, Rhodamine 110, Rhodamine 123, Rhodamine B, Rhodamine B200, Rhodamine BB, Rhodamine BG, Rhodamine B Extra, 5-Carboxytetramethylrhodamine (5-TAMRA), 5GLD, 6-Carboxyrhodamine 6G, Lisamin, Lisaminrhodamine B, Faricidine, Phalloidin, Rhodamine Red, Rhod-2, 6-Carboxy-X-Rhodamine (ROX), Carboxy-X-Rhodamine (5-ROX), Sulforodamine B can A derivative of an antibody, which may be selected from the group consisting of C, sulforhodamine G Extra, 6-carboxytetramethylrhodamine (TAMRA), tetramethylrhodamine (TRITC), rhodamine WT, Texas Red, and Texas Red-X.

10. An antibody according to any one of claims 1 to 9, comprising an effector portion bound thereto, wherein the effector portion is a therapeutic agent, a cytotoxic reagent, abrin A chain, anthracycline, amantine, α-amantine, auristatin, monomethyl auristatin E, monomethyl auristatin F, calicheamycin, camptothecin, combretastatin, crotin, cryptophycin, crucin, drastatin, duocalmycin, DNA alkylating agent, DNA repair inhibitor, duocalmycin, engine, exatecan or derivative thereof, DX-8951, exotoxin A chain, deruxtecan, diphtheria A chain, enomycin Antibodies may be selected from the group consisting of: geldanamycin, hemiastalin, inhibitors of ataxia telangiectasia and Rad3-related kinase, bezocertib, indolino-benzodiazepine dimers, meitansin, meitansinoid DM1, meitansinoid DM4, ozogamicin, phenomycin, prazienolide, plant toxins, puromycin, pyrrolobenzodiazepine dimers, lysine A chain, splicostatin, taxanes, toxins, tubulcin, tumor-activated prodrugs, topoisomerase inhibitors, vinca alkaloids, radiochemicals, radioisotopes, iodine-131, and yttrium-90.

11. The antibody according to claim 10, wherein the cytotoxic agent is duocalmycin.

12. The antibody according to claim 10 or 11, wherein a non-cleavable or cleavable linker is positioned between the antibody and the effector portion, and the cleavable linker releases the effector portion into or within the cell.

13. An isolated polynucleotide encoding an antibody according to any one of claims 1 to 12, or a combination comprising at least one polynucleotide having at least about 90% identity with any of these.

14. An expression vector comprising one or more polynucleotides as described in claim 13.

15. A host cell comprising the isolated polynucleotide described in claim 13 and / or the expression vector described in claim 14.

16. A composition comprising at least one antibody or derivative according to any one of claims 1 to 12, at least one isolated polynucleotide according to claim 13, or at least one expression vector according to claim 14, and / or at least one host cell according to claim 15, or a combination thereof, and a pharmaceutically acceptable carrier.

17. A method for detecting PTGFRN on cells and / or tissues, comprising contacting a test biological sample with an antibody or derivative described in any one of claims 1 to 13, and detecting the antibody bound to the biological sample or its components.

18. The method according to claim 17, further comprising comparing the amount of binding to the test biological sample or its components with the amount of binding to a control biological sample or its components, wherein the increase in binding to the test biological sample or its components compared with the control biological sample or its components indicates the presence of cells expressing PTGFRN in the test biological sample.

19. The method according to claim 17 or 18, wherein the test biological sample is mammalian cells, tissue, or biological fluid, and the biological fluid may be selected from the group consisting of blood, urine, plasma, serum, cerebrospinal fluid, saliva, and exosomes.

20. The method according to any one of claims 17 to 19, wherein the method is in vivo or in vitro.

21. An in vivo method for detecting tumor cells and / or diagnosing cancer, comprising administering at least one antibody, combination or derivative according to any one of claims 1 to 13, and detecting at least one of the antibodies bound to the tumor cells.

22. The method according to claim 21, wherein at least one of the antibodies or derivatives comprises at least one detectable label.

23. A method according to claim 22, comprising at least two antibodies and / or derivatives, each containing at least one detectable label, wherein the detectable labels of each antibody and / or derivative are the same or different.

24. The method according to any one of claims 21 to 23, comprising imaging a tumor for cancer-targeted treatment.

25. The method according to claim 22, further comprising treating the cancer.

26. The method according to claim 25, further comprising isolating the cancer cells or tissue and determining whether the cancer tissue overexpresses PTGFRN compared to non-cancerous cells, wherein PTGFRN expression may be determined by measuring the expression of the PTGFRN protein and / or RNA encoding PTGFRN.

27. A method for treating cancer, comprising administering to a mammal an antibody, combination, or derivative according to at least one of claims 1 to 13.

28. The method according to claim 27, wherein at least one of the antibodies or derivatives includes an effector portion.

29. The method according to claim 22, comprising at least two antibodies or derivatives, each containing at least one effector portion, wherein the effector portions of each antibody or derivative are the same or different.

30. The method according to any one of claims 27 to 29, further comprising isolating the cancer cells and determining whether the cancer tissue overexpresses PTGFRN compared to non-cancerous cells, wherein PTGFRN expression may be determined by measuring the expression of the PTGFRN protein and / or the RNA encoding PTGFRN.

31. A method for detecting, diagnosing and treating cancer, comprising imaging a tumor containing an antibody, combination, or derivative according to any one of claims 1 to 13, conjugated to the tumor, and targeting the tumor with a cancer treatment.

32. The method according to claim 27, wherein the antibody or derivative thereof includes a detectable label and / or effector portion.

33. The method according to claim 31 or 32, further comprising isolating the cancer cells and determining whether the cancer tissue overexpresses PTGFRN compared to non-cancerous cells, wherein PTGFRN expression may be determined by measuring the expression of the PTGFRN protein and / or the RNA encoding PTGFRN.

34. A method for treating, preventing and / or improving cancer in a mammal, comprising administering to the mammal at least one pharmaceutical composition in an effective dose of at least one antibody and / or derivative described in at least one of claims 1 to 13.

35. At least one antibody in at least one of the pharmaceutical compositions comprises a cytotoxic effector portion bound thereto, wherein the effector portion is a therapeutic agent, a cytotoxic reagent, abrin A chain, anthracycline, amantine, α-amantine, auristatin, monomethyl auristatin E, monomethyl auristatin F, calicheamycin, camptothecin, combretastatin, crotin, cryptophycin, crucin, drastatin, duocalmycin, DNA alkylating agent, DNA repair inhibitor, duocalmycin, engine, exatecan or its derivatives, DX-8951, exotoxin A chain, deruxtecan, diphtheria A chain, enomay The method according to claim 34, which may be selected from the group consisting of syn, geldanamycin, hemiastalin, inhibitors of ataxia telangiectasia and Rad3-related kinase, bezocertib, indolino-benzodiazepine dimers, meitansin, meitansinoid DM1, meitansinoid DM4, ozogamicin, phenomycin, prazienolide, plant toxins, puromycin, pyrrolobenzodiazepine dimers, lysine A chain, splicostatin, taxanes, toxins, tubulsin, tumor-activated prodrugs, topoisomerase inhibitors, vinca alkaloids, radiochemicals, radioisotopes, iodine-131 and yttrium-90.

36. The method according to claim 35, wherein the cytotoxic agent is duocalmycin.

37. The method according to claim 35 or 36, wherein a non-cleavable or cleavable linker is positioned between the antibody and the effector portion, and the cleavable linker releases the effector portion into or within the cell.

38. The method according to any one of claims 34 to 37, wherein the antibody is administered as an antibody-drug conjugate.

39. The method according to any one of claims 34 to 38, comprising administering at least two antibodies to the mammal, wherein at least one antibody is a naked antibody that does not contain a cytotoxic effector moiety bound thereto, and at least one antibody contains a cytotoxic effector moiety bound thereto.

40. The method according to any one of claims 34 to 38, wherein multiple doses are administered to the animal and / or the antibody is administered at a dose of about 1 to 50 mg / kg.

41. The method according to any one of claims 34 to 39, further comprising isolating the cancer cells and determining whether the cancer tissue overexpresses PTGFRN compared to non-cancerous cells, wherein the PTGFRN expression may be determined by measuring the expression of the PTGFRN protein and / or the RNA encoding PTGFRN.

42. The method according to any one of claims 34 to 41, wherein the mammal is a human.

43. The method according to any one of claims 34 to 42, wherein the cancer is selected from the group consisting of head and / or neck cancer, squamous cell carcinoma, epidermoid carcinoma, medulloblastoma, mesothelioma, and hematological cancer.

44. A kit for detecting the expression of PTGFRN in or on cells, tissues, or biological fluids, comprising an antibody or derivative according to any one of claims 1 to 13 and instructions for use.

45. The kit according to claim 41, wherein the antibody or derivative is present in a lyophilized form.

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