Polymorphisms as predictors of treatment response and overall survival

Abstract

The present disclosure relates to methods for determining whether a subject with cancer, specifically metastatic colorectal cancer, will respond to treatment using bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or predicting overall survival. The disclosure also relates to methods for selecting a treatment for cancer in a subject and compounds used in said treatment for cancer in said subject. The disclosure also relates to kits that can be used for these methods.

Description

[Technical Field]

[0001] The present disclosure relates to methods for determining whether a subject with cancer, specifically metastatic colorectal cancer, will respond to treatment using bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or predicting overall survival. The disclosure also relates to methods for selecting a treatment for cancer in a subject and compounds used in said treatment for cancer in said subject. The disclosure also relates to kits that can be used for these methods. [Background technology]

[0002] Colorectal cancer (CRC) is the second most commonly diagnosed cancer in women and the third most commonly diagnosed in men. It is estimated that 20% of newly diagnosed patients have distant metastatic disease at the time of diagnosis.

[0003] Significant increases in average median overall survival have been major advances in the treatment of metastatic colorectal cancer (mCRC) over the past 15 years, although clinical outcomes for patients remain highly variable.

[0004] Bevacizumab, a recombinant humanized IgG1 monoclonal antibody, is the first antiangiogenic agent approved for the treatment of mCRC. The mechanism of action of bevacizumab involves binding to circulating vascular endothelial growth factor A (VEGF-A) and blocking its binding to its receptors (VEGFR-1 and VEGFR-2) on the surface of endothelial cells, resulting in the inhibition of tumor angiogenesis, growth, and metastasis. The combination of bevacizumab with fluoropyrimidine chemotherapy agents is widely used as a first-line therapy for mCRC.

[0005] However, the clinical outcomes of bevacizumab monotherapy and combination therapy treatment are highly variable. Like most monoclonal antibodies, bevacizumab exhibits complex and variable pharmacokinetic and pharmacodynamic characteristics. This variability in patient outcomes is associated with several factors, including gender, body weight, tumor burden, binding to their molecular targets, and concurrent chemotherapy.

[0006] Although bevacizumab is widely used in oncology, validated predictors of treatment outcome remain lacking. Several clinical studies in patients with mCRC have specifically noted associations between the levels of biomarkers (e.g., lactate dehydrogenase, ICAM, E-selectin, endothelial nitric oxide synthase) and SNPs in genes involved in the angiogenesis pathway and response to bevacizumab. However, no validated biomarkers are currently available to guide patient selection for treatment with bevacizumab-based therapeutic agents.

[0007] The pathogenesis of mCRC involves the accumulation of genetic and epigenetic modifications in pathways controlling proliferation, apoptosis, and angiogenesis. Genomic variants of RAS and BRAF are important for the prognosis and prediction of mCRC. KRAS genomic variants at either codon 12 or codon 13 are identifiable in 12–75% of CRCs, and these genomic variants are independently associated with poor prognosis. NRAS mutations are also associated with poor prognosis. Similarly, the most frequent BRAF activating mutation at codon 600 (V600E) is present in less than 10% of tumors and is a strong negative prognostic marker.

[0008] Single nucleotide polymorphisms (SNPs) in genes involved in VEGF-dependent and -independent angiogenesis pathways, as well as other major intracellular signaling pathways involved in the pathogenesis of mCRC, have been investigated. Specifically, the VEGF-A gene and the I gene, which have been shown to be associated with mCRC, have been investigated. Polymorphisms in the CAM-1 gene have been identified (see Zhang et al., Med. Sci. Monit., 2016, 22:569-579; Wether et al., Eur. J. Surg. Oncol., 2000, 26:657-662, each of which is incorporated herein by reference). The role of VEGF-A SNPs on the therapeutic efficacy of bevacizumab-based therapy has been investigated in many tumors. For example, the genotype distribution of the VEGF-A gene rs699947 polymorphism was analyzed in bevacizumab-based therapy for colorectal cancer. However, no significant correlation of the VEGF-A gene rs699947 polymorphism with response to therapy was determined (see Cui et al., Oncotarget, 2017, 8(62):105472-104478, each of which is incorporated herein by reference).

[0009] Many studies have focused on PFS and OS; however, these studies have been small and have produced mixed results. There have been no statistically significant findings linking polymorphisms in the VEGF-A and ICAM-1 genes to improved clinical response and / or prolonged overall survival when patients are treated with bevacizumab in combination with fluoropyrimidine chemotherapy.

[0010] There is a need to identify the subpopulation or subpopulations of mCRC patients who would benefit from bevacizumab treatment, thereby optimizing the management of their disease. Specifically, validated biomarkers to guide patient selection are needed for bevacizumab in combination with fluoropyrimidine chemotherapy. Biomarkers that have been demonstrated to be statistically significant for increased overall survival are needed for the combination of bevacizumab with fluoropyrimidine chemotherapy. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] Zhang et al., Med. Sci. Monit., 2016, 22:569-579 [Non-patent document 2] Wether et al., Eur. J. Surg. Oncol., 2000, Volume 26: 657-662. [Non-patent document 3] Cui et al., Oncotarget, 2017, Volume 8 (No. 62), pp. 105472-104478 [Non-patent document 4] "Guideline for similar biological medicinal products containing monoclonal antibodies regarding non-clinical and clinical issues" (https: / / www.ema.europa.eu / en / documents / scientific-guideline / guideline-similar-biological-medicinal-products-containing-monoclonal-antibodies-non-clinical_en.pdf) [Non-Patent Document 5] "Scientific Considerations for Demonstrating Bioequivalence to a Reference Product" (https: / / www.fda.gov / media / 82647 / download) [Non-patent document 6] Eisenhauer et al., European Journal of Cancer, Vol. 45, 2009, pp. 228-247 [Non-Patent Document 7] Bland et al., BMJ 2004, 328:1073 [Non-patent document 8] Cutsem et al., Ann Oncol., August 2016, Volume 27 (No. 8): pp. 1386-422 (https: / / www.ncbi.nlm.nih.gov / pubmed?term=27380959) [Non-Patent Document 9] NCCN Guidelines for Colorectal Cancer (https: / / www.nccn.org / professionals / physician_gls / default.aspx) [Non-Patent Document 10] DNA Sequencing by Capillary Electrophoresis, Applied Biosystems Chemistry Guide, 2nd Edition, https: / / tools.thermofisher.com / content / sfs / manuals / cms_041003.pdf Summary of the Invention

[0012] Representative features of the present invention are set forth in the following clauses, which may be effective alone or in any combination with one or more features disclosed in the text and / or drawings of this specification.

[0013] 1. A method for determining whether a subject with metastatic colorectal cancer (mCRC) will respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or for predicting the overall survival (OS) of said subject, comprising: (a) determining the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in a sample obtained from the subject, or identifying a subject identified as having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism; and (b) indicating that if at least one of the VEGF-A rs699947 polymorphism and the ICAM-1 rs1799969 polymorphism is present, the subject is likely to respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or is likely to have a longer overall survival when treated with bevacizumab and a fluoropyrimidine chemotherapeutic agent. The method comprising or consisting of the steps of:

[0014] 2. The method of clause 1, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0015] 3. The method of any one of clauses 1 or 2, wherein the presence or absence of said allele is determined by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0016] 4. The method of any one of clauses 1 to 3, wherein the fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0017] 5. The method of any one of clauses 1 to 3, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) or capecitabine / oxaliplatin (BEV-CapOX).

[0018] 6. The method of any one of clauses 1 to 5, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0019] 7. The method of any one of clauses 1 to 5, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0020] 8. The patient is in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelveth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, or eighteenth cycle. 8. The method of any one of clauses 1-7, having 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0021] 9. The method of any one of clauses 1 to 8, wherein the presence of the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism in the sample obtained from the subject, or identifying a subject identified as having the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism, indicates that the subject is also likely to respond to maintenance treatment and / or may also have a greater likelihood of overall survival with maintenance treatment.

[0022] 10. The method of any one of clauses 1-9, wherein the patient has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0023] 11. The method of any one of clauses 1 to 10, wherein there is no maintenance treatment.

[0024] 12. The method of any one of clauses 9 to 10, wherein said maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0025] 12A. Subjects having the VEGF-A rs699947 polymorphism have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months when treated with bevacizumab and a fluoropyrimidine chemotherapy agent; or 13. The method of any one of clauses 1-12, wherein a subject having the rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0026] 12B. Subjects having the ICAM-1 rs1799969 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months, or 13. The method of any one of clauses 1-12, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0027] 13. A method for selecting a treatment for metastatic colorectal cancer in a subject, comprising: (a) determining the presence or absence of the VEGF-A rs699947 mutation and / or the ICAM-1 rs1799969 mutation in a sample obtained from the subject, or identifying a subject identified as having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism; and (b) selecting the treatment comprising, consisting essentially of, or consisting of bevacizumab and a fluoropyrimidine chemotherapeutic agent when at least one of a VEGF-A rs699947 mutation and an ICAM-1 rs1799969 mutation is present. The method comprising, consisting essentially of, or consisting of the steps of:

[0028] 14. The method of clause 13, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0029] 15. The method of any one of clauses 13 or 14, wherein the presence or absence of the allele is determined by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0030] 16. The method of any one of clauses 13 to 15, wherein the fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0031] 17. The method of any one of clauses 13 to 15, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0032] 18. The method of any one of clauses 13 to 17, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0033] 19. The method of any one of clauses 13 to 17, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0034] 20. The method of any one of clauses 13-19, wherein the patient has received 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0035] 21. The presence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in the sample obtained from the subject or the VEGF-A 21. The method of any one of clauses 13 to 20, wherein identifying a subject identified as having the rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism indicates that said subject is also likely to respond to maintenance treatment and / or that maintenance treatment may also improve overall survival.

[0036] 22. The patient is on cycle 1, 2, 3, 4, 5, or 6 22. The method of any one of clauses 13-21, comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0037] 23. The method according to any one of clauses 13 to 20, wherein there is no maintenance treatment.

[0038] 24. The method of any one of clauses 21-22, wherein said maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0039] 24A. Subjects having the VEGF-A rs699947 polymorphism have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months when treated with bevacizumab and a fluoropyrimidine chemotherapy agent; or 25. The method of any one of clauses 13-24, wherein a subject having the rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0040] 24B. Subjects having the ICAM-1 rs1799969 polymorphism have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months when treated with bevacizumab and a fluoropyrimidine chemotherapy agent, or 25. The method of any one of clauses 13-24, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0041] 25. A method of treating metastatic colorectal cancer in a subject, comprising: (a) determining the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in a sample obtained from the subject, or identifying a subject identified as having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism; and (b) administering bevacizumab and a fluoropyrimidine chemotherapeutic agent when at least one of the VEGF-A rs699947 polymorphism and the ICAM-1 rs1799969 polymorphism is present; The method comprising, consisting essentially of, or consisting of the steps of:

[0042] 26. The method of clause 25, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0043] 27. The presence or absence of the allele can be determined by PCR, PCR-RFLP, direct sequencing, or T 27. The method of any one of clauses 25 or 26, wherein the gene is identified by aqMan, and / or next generation signaling.

[0044] 28. The method of any one of clauses 25 to 27, wherein the fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0045] 29. The method of any one of clauses 25 to 27, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0046] 30. The method of any one of clauses 25 to 29, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0047] 31. The method of any one of clauses 25 to 29, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0048] 32. The method of any one of clauses 25-31, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0049] 33. The presence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in the sample obtained from the subject or the VEGF-A 33. The method of any one of clauses 25 to 32, wherein identifying a subject identified as having the rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism indicates that said subject is also likely to respond to maintenance treatment and / or that maintenance treatment may also improve overall survival.

[0050] 34. The method of any one of clauses 25-33, wherein the patient has received 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0051] 35. The method according to any one of clauses 25 to 32, wherein there is no maintenance treatment.

[0052] 36. The method of any one of clauses 33-34, wherein said maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0053] 36A. Subjects having the VEGF-A rs699947 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months; or 37. The method of any one of clauses 25-36, wherein a subject having the rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0054] 36B. Subjects having the ICAM-1 rs1799969 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months, or 37. The method of any one of clauses 25-36, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0055] 37. A combination of bevacizumab and a fluoropyrimidine chemotherapy agent, (a) determining the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in a sample obtained from a subject, or Identifying subjects identified as having the rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism; and (b) administering bevacizumab and a fluoropyrimidine chemotherapeutic agent when at least one of the VEGF-A rs699947 polymorphism and the ICAM-1 rs1799969 polymorphism is present; said combination for use in a method for treating metastatic colorectal cancer in said subject, comprising:

[0056] 38. The method of clause 37, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0057] 39. The method of any one of clauses 37 or 38, wherein the presence or absence of the allele is determined by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0058] 40. The method of any one of clauses 37 to 39, wherein the fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0059] 41. The method of any one of clauses 37 to 39, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0060] 42. The bevacizumab is administered at a dose of 2.5 to 7.5 mg / kg once every two weeks, preferably 42. The method of any one of clauses 37 to 41, wherein said fluoropyrimidine chemotherapeutic agent is administered as an intravenous infusion at a dose of 5 mg / kg in combination with said fluoropyrimidine chemotherapeutic agent.

[0061] 43. The method of any one of clauses 37 to 41, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0062] 44. The method of any one of clauses 37 to 43, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0063] 45. The presence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in the sample obtained from the subject or the VEGF-A 45. The method of any one of clauses 37 to 44, wherein identifying a subject identified as having the rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism indicates that said subject is also likely to respond to maintenance treatment and / or that maintenance treatment may also improve overall survival.

[0064] 46. ​​The method of any one of clauses 37-45, wherein the patient has received 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0065] 47. The method according to any one of clauses 37 to 44, wherein there is no maintenance treatment.

[0066] 48. The method of any one of clauses 45-46, wherein the maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0067] 48A. Subjects having the VEGF-A rs699947 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months; or 49. The method of any one of clauses 37-48, wherein a subject having the rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0068] 48B. Subjects with the ICAM-1 rs1799969 polymorphism were treated with bevacizumab and or a subject having said ICAM-1 rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0069] 49. A method for treating metastatic colorectal cancer in a subject having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism, the method comprising administering bevacizumab and a fluoropyrimidine chemotherapeutic agent.

[0070] 50. The method of clause 49, wherein the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism can be identified using a sample obtained from the subject, and the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0071] 51. The method of any one of clauses 49 or 50, wherein the presence or absence of the allele is determined by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0072] 51A. The subject has the VEGF-A rs699947 polymorphism and / or ICAM-1 51. The method of any one of clauses 49 or 50, wherein the subject is identified as having the rs1799969 polymorphism.

[0073] 52. The method of any one of clauses 49-51A, wherein said fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0074] 53. The method of any one of clauses 49 to 51A, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0075] 54. The method of any one of clauses 49 to 53, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0076] 55. The method of any one of clauses 49 to 53, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0077] 56. The method of any one of clauses 49 to 55, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles. How to do it.

[0078] 57. The method of any one of clauses 49 to 56, wherein the presence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in the sample obtained from the subject, or identification of the subject as having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism, indicates that the subject is also likely to respond to maintenance treatment and / or that maintenance treatment may also improve overall survival.

[0079] 58. The method of any one of clauses 49-57, wherein the patient has received 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0080] 59. The method of any one of clauses 49 to 56, wherein there is no maintenance treatment.

[0081] 60. The method of any one of clauses 57-58, wherein said maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0082] 60A. Subjects having the VEGF-A rs699947 polymorphism have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months when treated with bevacizumab and a fluoropyrimidine chemotherapy agent; or 61. The method of any one of clauses 49-60, wherein a subject having the rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0083] 60B. Subjects having the ICAM-1 rs1799969 polymorphism treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months, or 61. The method of any one of clauses 49-60, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0084] 61. A combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent for use in a method for treating metastatic colorectal cancer in a subject having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism.

[0085] 62. The method of clause 61, wherein the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism can be identified using a sample obtained from the subject, and the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0086] 63. The method of any one of clauses 61 or 62, wherein the presence or absence of the allele is determined by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0087] 63A. The subject has the VEGF-A rs699947 polymorphism and / or ICAM-1 51. The method of any one of clauses 49 or 50, wherein the subject is identified as having the rs1799969 polymorphism.

[0088] 64. The method of any one of clauses 61-63A, wherein said fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0089] 65. The method of any one of clauses 61-63A, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0090] 66. The method of any one of clauses 61 to 65, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0091] 67. The method of any one of clauses 61 to 65, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0092] 68. The method of any one of clauses 61 to 67, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0093] 69. The method of any one of clauses 61 to 68, wherein the presence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in the sample obtained from the subject, or identification of the subject as having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism, indicates that the subject is also likely to respond to maintenance treatment and / or that maintenance treatment may also improve overall survival.

[0094] 70. The method of any one of clauses 61-69, wherein the patient has received 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0095] 71. The method according to any one of clauses 61 to 68, in which there is no maintenance treatment.

[0096] 72. The method of any one of clauses 69-70, wherein the maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0097] 72A. Subjects having the VEGF-A rs699947 polymorphism have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months when treated with bevacizumab and a fluoropyrimidine chemotherapy agent; or 73. The method of any one of clauses 61-72, wherein a subject having the rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0098] 72B. Subjects having the ICAM-1 rs1799969 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months, or 73. The method of any one of clauses 61-72, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0099] 73. A method of treating cancer in a subject with metastatic colorectal cancer (mCRC), comprising: administering bevacizumab and a fluoropyrimidine chemotherapeutic agent to the subject; and determining the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in a sample obtained from the subject, or Identifying subjects identified as having the rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism. The method comprising or consisting of the steps of:

[0100] 74. The method of clause 73, wherein the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism can be identified using a sample obtained from the subject, and the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0101] 75. The method of any one of clauses 73 or 74, wherein the presence or absence of the allele is determined by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0102] 75A. The subject has the VEGF-A rs699947 polymorphism and / or ICAM-1 any one of clauses 73 or 75, identified as having the rs1799969 polymorphism The method described below.

[0103] 75B. The method of any one of clauses 73-75A, wherein the fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0104] 76. The method of any one of clauses 73-75A, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0105] 77. The method of any one of clauses 73 to 76, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0106] 78. The method of any one of clauses 73 to 76, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0107] 79. The method of any one of clauses 73-78, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0108] 80. The method of any one of clauses 73-79, wherein the patient has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0109] 81. The method according to any one of clauses 73 to 79, in which there is no maintenance treatment.

[0110] 82. The method of any one of clauses 80-81, wherein said maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0111] 82A. Subjects having the VEGF-A rs699947 polymorphism treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months; or subjects having the VEGF-A rs699947 polymorphism treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33, 44, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, or 83. The method of any one of clauses 73-82, wherein the method results in an OS of 5 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0112] 82B. Subjects having the ICAM-1 rs1799969 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months; or 61. The method of any one of clauses 49-60, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months.

[0113] 83. A method for determining the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism in a subject, comprising: (a) collecting a whole blood sample; (b) separating the blood and isolating a serum sample; (c) extracting genomic DNA from blood leukocytes; (d) measuring the DNA concentration; (e) analyzing genomic variants by PCR and purifying the PCR products; (f) performing our DNA sequence analysis on the purified PCR products; or (g) adding a label that binds to the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism and detecting the bound label; The method comprising:

[0114] 84. A method for treating metastatic colorectal cancer in a subject, the method comprising administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject known to have the ICAM-1 rs1799969G / A polymorphism.

[0115] 85. A method for administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject with metastatic colorectal cancer, the method comprising determining the presence or absence of the ICAM-1 rs1799969G / A polymorphism in a sample obtained from the subject, and administering the combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to the subject found to have the ICAM-1 rs1799969G / A polymorphism.

[0116] 86. A method of treating metastatic colorectal cancer in a subject, the method comprising administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject known to have the VEGF-A rs699947A / A polymorphism.

[0117] 87. A method for administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject having metastatic colorectal cancer, the method comprising determining the presence or absence of the VEGF-A rs699947A / A polymorphism in a sample obtained from the subject, and administering the combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to the subject found to have the VEGF-A rs699947A / A polymorphism.

[0118] 88. The method of any one of clauses 84 to 87, wherein said combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent is administered as first-line therapy.

[0119] 89. The method of any one of clauses 84 to 88, wherein said identifying step comprises detecting said polymorphism by a method comprising one or more of PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next-generation sequencing.

[0120] 90. The method of any one of clauses 84 to 89, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

[0121] 91. The method of any one of clauses 84 to 90, wherein the fluoropyrimidine chemotherapeutic agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI) or 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX).

[0122] 92. The method of any one of clauses 84 to 90, wherein the fluoropyrimidine chemotherapeutic agent is capecitabine / irinotecan (BEV-CapIRI) and / or capecitabine / oxaliplatin (BEV-CapOX).

[0123] 93. The method of any one of clauses 84 to 92, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg, preferably at a dose of 5 mg / kg, once every two weeks.

[0124] 94. The method of any one of clauses 84 to 92, wherein said bevacizumab is administered as an intravenous infusion in combination with said fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks, preferably at a dose of 7.5 mg / kg in a cycle of every three weeks.

[0125] 95. The method of any one of clauses 84-94, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 treatment cycles.

[0126] 96. The method of any one of clauses 84-95, wherein the patient has had 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 maintenance treatment cycles.

[0127] 97. The method according to any one of clauses 84 to 95, in which there is no maintenance treatment.

[0128] 98. The method of clause 96, wherein the maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy.

[0129] 98A. Subjects with the VEGF-A rs699947 polymorphism treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 45 to 65 months, 45 to 60 months, or 50 to 55 months. or the method of any one of clauses 84 to 98, wherein a subject having the VEGF-A rs699947 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 44 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

[0130] 98B. Subjects having the ICAM-1 rs1799969 polymorphism who are treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months; or 99. The method of any one of clauses 84-98, wherein subjects having the rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months. [Brief explanation of the drawings]

[0131] Embodiments of the present invention will now be described with reference to the accompanying drawings.

[0132] [Figure 1] Figure 1 shows OS according to VEGF-A rs699947 polymorphism. Median OS was 18.1 months in C / C carriers compared with 52 months in A / A carriers (p=0.043).

[0133] [Figure 2] Figure 1 shows OS according to the ICAM-1 rs1799969 polymorphism. Median OS was 29.1 months in G / G carriers versus 48.7 months in G / A carriers (p=0.036).

[0134] [Figure 3] Figure 1 shows PFS according to the VEGF-A rs699947 polymorphism. The median PFS was 10.1 months in C / C carriers and 31.1 months in A / A carriers (P=0.006). DETAILED DESCRIPTION OF THE INVENTION

[0135] The following description examples illustrate various embodiments of the present disclosure in detail. Those skilled in the art will appreciate that there are numerous variations and modifications that fall within the scope of this disclosure. Therefore, the description of the disclosed embodiments should not be considered as limiting the scope of the present disclosure.

[0136] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. All patents, patent applications, patent application publications, and other publications referenced herein are incorporated by reference in their entirety unless otherwise indicated. In the event that there are multiple definitions for a term herein, those in this section prevail unless otherwise indicated.

[0137] "Bevacizumab" refers to the compound Avastin®, a recombinant humanized monoclonal IgG1 antibody that binds to and inhibits the biological activity of human vascular endothelial growth factor A (VEGF-A). The interaction of VEGF with its receptor results in endothelial cell proliferation and angiogenesis in in vitro models of angiogenesis. Bevacizumab binds to circulating VEGF-A and It blocks VEGFR-A from binding to its receptors (VEGFR-1 and VEGFR-2) on the surface of endothelial cells, resulting in the inhibition of tumor angiogenesis, growth, and metastasis. "Bevacizumab" encompasses all applicable anti-VEGF antibodies or anti-VEGF antibody fragments that meet the requirements necessary to obtain marketing authorization as identical or biosimilar products to the reference medicinal product / reference drug, bevacizumab (Avastin®), from regulatory authorities such as the European Medicines Agency (EMA) and the U.S. Food and Drug Administration (FDA). See the EMA's "Guideline for Monoclonal Antibody-Containing Follow-On Biological Medicinal Products Regarding Nonclinical and Clinical Issues" (https: / / www.ema.europa.eu / en / documents / scientific-guideline / guideline-similar-biological-medicinal-products-containing-monoclonal-antibodies-non-clinical_en.pdf). See the FDA's "Scientific Considerations for Demonstrating Bioequivalence to the Reference Product" (https: / / www.fda.gov / media / 82647 / download). Biosimilars have been approved as follow-on biologics to the reference drug / product bevacizumab (Avastin®). Examples include, but are not limited to, Mvasi (ABP-215) or Mvasi (bevacizumab-awwb) (Amgen) and Zirabev (Pfizer). Mvasi has been approved by the EMA and FDA as a biosimilar to the reference drug / product bevacizumab (Avastin®). Zirabev has been approved by the EMA as a biosimilar to the reference drug / product bevacizumab (Avastin®). These biosimilars bind to and inhibit the biological activity of human vascular endothelial growth factor A (VEGF-A). Biosimilars are approved according to the same standards of pharmaceutical quality, safety, and efficacy that apply to all biological drugs.

[0138] Examples of anti-cancer drugs include, but are not limited to, BCD-021 (Biocad, Russia), FKB238 (AstraZeneca / Kyowa Hakko Kirin Fujifilm Biologics, USA / Japan), BCD500 (BIOCND, Korea), Krabeva (Biocon, India*), BI695502 (Boehringer Ingelheim, Germany), CT-P16 (Celltrion, Korea), CHS-5217 (Cohilus, USA), DRZ_BZ (Dr. Reddy's Laboratories, India), Cizumab (Hetero (Lupin), India), Bevax (mAbxience, Spain / Argentina), ONS-1045 (Oncobiolo A number of biosimilars are also in development, such as Zyx Pharmaceuticals / Viropro, USA), PF-06439535 (Pfizer, USA), HD204 (Prestige Biopharmaceuticals, Singapore), Bevacirel (Reliance Life Sciences / Lupin, India), and SB8 (Samsung Bioepis (Biogen / Samsung) / Merck, Korea / USA), which would be biosimilars according to the present invention if the appropriate regulatory authorities, such as the European Medicines Agency (EMA) or the US Food and Drug Administration (FDA), approve these compounds as biosimilars to bevacizumab (Avastin®).

[0139] The bevacizumab treatment can be in the form of a monotherapy or a combination therapy with a chemotherapeutic agent, particularly in the form of a combination therapy with a fluoropyrimidine chemotherapeutic agent as described herein. For example, the bevacizumab treatment can be a first-line therapy consisting of a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent, followed by a maintenance therapy consisting of bevacizumab monotherapy.

[0140] As used herein, "metastatic colorectal cancer (mCRC)" refers to a subject with metastatic cancer of the colon or rectum. In mCRC, the cancer has spread from the colon or rectum to organs and tissues distal to the colon or rectum. Colorectal cancer most often spreads to the liver, but may also spread to the lungs, bone, brain, spinal cord, peritoneum, and other organs. It may also spread to other places, such as the lining of the abdominal cavity or distant lymph nodes.

[0141] As used herein, "VEGF-A" is a member of the VEGF family. Vascular endothelial growth factor A (VEGF-A) is a protein encoded by the VEGFA gene. VEGF-A binds with high affinity to its receptors VEGFR-1 and VEGFR-1, promoting angiogenic signaling. VEGF-A is considered a major activator of angiogenesis. It acts selectively on vascular endothelial cells and stimulates both normal and abnormal angiogenesis.

[0142] As used herein, "ICAM-1" is known to be a member of the immunoglobulin gene superfamily of adhesion molecules. Intercellular adhesion molecule 1 (ICAM-1) is a cell surface glycoprotein typically expressed on endothelial cells and immune system cells. This adhesion molecule binds to integrins of the CD11a / CD18 or CD11b / CD18 type. ICAM-1 expression is closely associated with cancer metastasis.

[0143] As used herein, "single nucleotide polymorphism (SNP)" refers to a single base substitution occurring at a specific position in the genome. For example, at a specific base position in the human genome, a C nucleotide may be found in most individuals, but in a small number of individuals, the position is occupied by A. This means that an SNP exists at this specific position, and the two possible nucleotide variations, C or A, can be said to be the alleles at this position. VEGF-A rs699947 polymorphism and ICAM-1 rs1799969 polymorphism are SNPs.

[0144] As used herein, "VEGF-A rs699947 polymorphism," "VEGF-A rs699947," "VEGF-A rs699947 polymorphism," and "VEGF-A rs699947 variant" refer to a single nucleotide polymorphism at position 2578 of the VEGF-A gene. A subject can be a carrier of the A / A allele or the C / C allele. "VEGF-A rs699947 allele A / A" refers to a subject who has the allele A / A at position 2578 of the VEGF-A gene, which is a SNP and is referred to as a VEGF-A polymorphism. "VEGF-A rs699947 allele C / C" refers to a subject who has the allele C / C at position 2578 of the VEGF-A gene, which is the wild-type form of the gene.

[0145] As used herein, "ICAM-1 rs1799969 polymorphism," "ICAM-1 rs1799969," "ICAM-1 rs1799969 gene polymorphism," and "ICAM-1 rs1799969 variant" refer to a single nucleotide polymorphism at position 241 of the ICAM-1 gene. A subject can be a carrier of the G / G allele or the G / A allele. "ICAM-1 rs1799969 allele G / A" refers to a subject who has the allele G / A at position 241 of the ICAM-1 gene, which is a SNP and is referred to as an ICAM-1 polymorphism. "ICAM-1 rs1799969 allele G / G" refers to a subject who has the allele G / G at position 241 of the ICAM-1 gene, which is the wild-type form of the gene.

[0146] As used herein, "fluoropyrimidine chemotherapeutic agents" refers to a group of substances used to treat cancer. Fluoropyrimidines are a type of antimetabolite, and examples include capecitabine, floxuridine, and fluorouracil (5-FU). In some fluoropyrimidine chemotherapeutic agents, the fluoropyrimidine is administered with oxaliplatin and / or irinotecan. Examples include 5-fluorouracil / leucovorin / irinotecan (FOLFIRI), 5-fluorouracil / leucovorin / oxaliplatin (FOLFOX), capecitabine / irinotecan (CapIRI) or capecitabine / oxaliplatin (CapOX), and 5-fluorouracil / leucovorin / oxaliplatin / irinotecan (FOLFOXIRI).

[0147] As used herein, "progression-free survival" or "PFS" refers to the length of time a patient lives with a disease, such as cancer, without the disease worsening during and after treatment. In clinical trials, measuring progression-free survival is one way to see how well a new treatment is working. OS is a different way to see how well a new treatment is working. Progression-free survival is not linked to OS. PFS cannot be used to predict OS outcomes, and there is no relationship between these two endpoints.

[0148] As used herein, "likely to respond to treatment" refers to an increased probability of responding to treatment using a combination of bevacizumab and a fluoropyrimidine chemotherapy agent compared to a subject or group of subjects who do not possess the relevant polymorphism. For example, subjects with the VEGF-A rs699947 polymorphism are more likely to respond to treatment using bevacizumab and a fluoropyrimidine chemotherapy agent, increasing the probability of responding to treatment compared to a subject or group of subjects with wild-type VEGF-A rs699947. For example, subjects with the ICAM-1 rs1799969 polymorphism are more likely to respond to treatment using bevacizumab and a fluoropyrimidine chemotherapy agent, increasing the probability of responding to treatment compared to a subject or group of subjects with wild-type ICAM-1 rs1799969. A subject is responsive to treatment if the subject's life expectancy is extended beyond the life expectancy predicted if no treatment is administered. A subject is responding to a treatment if the subject has an increased overall survival compared to the overall survival if no treatment is administered. RECIST criteria can be used to determine whether a patient is responding to treatment and / or clinical evaluation.

[0149] As used herein, "overall survival" or "OS" refers to the time measured from the start of treatment to death, or from the end of treatment to death, or from a change in treatment to death. In clinical trials, measuring overall survival is one way to see how well a new treatment is working. Overall survival refers to the probability of survival at a specific time point between the start of treatment and death, for example, as represented by a Kaplan-Meier plot. Survival analysis was performed using Cox regression and Kaplan-Meier plot, which are the most widely used statistical tools for survival data analysis. Additionally, the log-rank test was used to check the null hypothesis and confirm significance. These statistical tools are well known and widely used by those skilled in the art.

[0150] As used herein, " RECIST " refers to " Response Evaluation Criteria in Solid Tumors ", which is a published set of rules that defines when cancer patients show favorable response during treatment.This response is evaluated according to the RECIST criteria version 1.1, published by Eisenhauer et al., European Journal of Cancer, Vol. 45, 2009, pp. 228-247, which is incorporated herein by reference.RECIST criteria is a standard method for those skilled in the art.

[0151] As used herein, "ECOG performance status" is a standard measure for measuring the extent to which a disease affects a patient's ability to perform daily activities (known to physicians and researchers as the patient's performance status). The ECOG performance status scale describes a patient's level of function with respect to their ability to care for themselves, their daily activities, and their physical abilities (walking, working, etc.). The scoring scale is a way to define the patient population being studied in a clinical trial so that the definition can be uniformly reproduced among physicians enrolling patients. The scale was developed by the Eastern Cooperative Oncology Group (ECOG), now part of the ECOG-ACRIN Cancer Research Group, and published in 1982. The scale is shown below:

[0152] [Table 1]

[0153] As used herein, "sample" refers to any sample capable of providing genomic DNA for analysis to determine the presence or absence of the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism. These samples are well known in the art and include, but are not limited to, whole blood, serum, peripheral blood leukocytes, or saliva.

[0154] As used herein, "associated with prolonged OS" or "associated with prolonged OS" means that the polymorphism is found in patients who are more likely to respond to treatment with bevacizumab and a fluoropyrimidine-based chemotherapy agent and / or who experience longer overall survival with the treatment than patients with a different polymorphism. For example, patients with the ICAM-1 rs1799969 polymorphism (G / A allele) are more likely to respond to treatment with bevacizumab and a fluoropyrimidine-based chemotherapy agent than patients with the wild-type ICAM-1 rs1799969 (G / G allele). For example, patients with the VEGF-A rs699947 polymorphism (A / A allele) are more likely to respond to treatment with bevacizumab and a fluoropyrimidine-based chemotherapy agent than patients with the wild-type ICAM-1 rs1799969 (C / C allele). The association is statistically significant.

[0155] As used herein, "statistical significance," "statistically significant," "significant," or "significantly" refers to the possibility that the relationship between two or more variables is caused by something other than chance. Statistical hypothesis testing is used to determine whether the results of a data set are statistically significant. This test provides a p-value, which represents the likelihood that the results can be explained by chance. A p-value of 5% or less (p<0.05) is considered statistically significant. For example, the VEGF-A rs699947 polymorphism (allele A / A) is significantly associated with prolonged OS compared to wild-type VEGF-A rs699947 (allele C / C) (p=0.043, i.e., less than 5%). The ICAM-1 rs1799969 polymorphism (allele G / A) is significantly associated with prolonged OS compared to wild-type ICAM-1 rs1799969 (allele G / G) (p=0.036, i.e., less than 5%).

[0156] As used herein, "log-rank test" refers to a hypothesis test for comparing the survival distributions of two samples. The log-rank test is used to compare the time to an event (from first treatment to death). It is widely used in clinical trials to demonstrate the efficacy of new treatments compared to control treatments, where a measure of response (such as time to response) is measured. This test is well known to those skilled in the art and is described in Bland et al., BMJ 2004, 328:1073, which is incorporated herein by reference. The log-rank test is more sophisticated than other hypothesis tests, such as ANOVA, which only perform a simple analysis of raw data, and can account for non-normality in the study population.

[0157] As used herein, "Cox regression" refers to a method for examining the effect of several variables on the occurrence of a specific event. In the context of outcomes such as death, this is known as Cox regression for survival analysis. To address any limitations in trial feasibility at the time of recruitment, we calculated the effect size that the study could capture for a given number of patients. With an 80% detection value and an α set at 0.05, we detected at least a 1.5-year difference in median PFS and OS between different polymorphisms for 46 patients with a 60-month follow-up period and 1-year recruitment period.

[0158] The term "subject" refers to mammals, such as humans, domestic animals, such as cats or dogs, livestock, including but not limited to, cows, horses, goats, sheep, and pigs, wild animals (wild or zoo), laboratory animals, such as mice, rats, rabbits, goats, sheep, pigs, dogs, and cats, birds, such as chickens, turkeys, and songbirds. The subject may be a child, such as an adolescent, or an adult. Preferably, the subject is an adult.

[0159] The term "treatment" refers to any treatment of a disease in a subject, e.g., a mammal, particularly a human, and includes (i) preventing the disease and / or reducing the risk of the disease in a subject who may be predisposed to the disease but has not yet been diagnosed with the condition, thereby constituting prophylactic treatment for the disease, and includes (ii) inhibiting the onset and / or slowing the rate of onset of the disease, e.g., arresting its development, (iii) alleviating the disease, e.g., causing regression of the disease, or (iv) alleviating the symptoms mediated by the disease or the symptoms of the disease. Treatment of subjects who have previously undergone and / or are currently undergoing and / or are about to undergo treatment for cancer is contemplated herein.

[0160] The term "therapeutically effective amount" refers to an amount of a compound, e.g., bevacizumab and / or a fluoropyrimidine chemotherapeutic agent, that is sufficient to effect treatment when administered to a subject in need of such treatment. The therapeutically effective amount varies depending on the subject and disease being treated, the subject's weight and age, the severity of the disease state, the method of administration, etc., and can be easily determined by one skilled in the art.

[0161] The term "consisting essentially of" means that other specified components may be present, i.e., components that do not materially affect the basic characteristics of the compound or composition.

[0162] When a range of values ​​is stated, it is understood that the upper and lower limits of that range, and each intervening value between those limits, are encompassed within the embodiment.

[0163] We investigated the role of SNPs in the angiogenic genes VEGF-A and ICAM-1, as well as KRAS, NRAS, and BRAF genes, to predict clinical outcomes and tumor response in patients with mCRC treated with bevacizumab in combination with fluoropyrimidine chemotherapy. Genes and polymorphisms known to regulate proliferation, apoptosis, and VEGF-induced or non-VEGF-induced angiogenesis were selected.

[0164] The present invention relates to the VEGF-A rs699947 polymorphism (allele A / A) and ICAM-1 This study is based on the discovery that the SNP rs1799969 polymorphism (allele G / A) is a predictor of the likelihood that patients with mCRC will respond to the combination of bevacizumab and a fluoropyrimidine chemotherapy agent. The VEGF-A rs699947 polymorphism is significantly associated with prolonged OS compared to wild-type VEGF-A rs699947 (p=0.043). The ICAM-1 rs1799969 polymorphism is significantly associated with prolonged OS compared to ICAM-1 rs1799969 allele G / G (p=0.036).

[0165] The statistically significant finding that subjects carrying the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism have a prolonged OS compared to subjects who do not carry these polymorphisms (i.e., carry the wild-type alleles) identifies a new patient population that will derive long-term benefit from treatment with bevacizumab in combination with a fluoropyrimidine chemotherapeutic agent.

[0166] This finding means that those subjects carrying the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism may respond and / or have a prolonged overall survival (OS) when treated with bevacizumab and a fluoropyrimidine chemotherapy agent.

[0167] This discovery allows physicians to select treatments for mCRC patients based on the presence or absence of the SNPs VEGF-A rs699947 and ICAM-1 rs1799969 in the genomic DNA of the patients. This provides financial benefits to insurance companies or healthcare systems such as the NHS. For example, bevacizumab is not approved in the UK due to its lack of cost-effectiveness, as only a small number of patients benefit from adding it to chemotherapy. However, this study shows that approximately 27% of patients with the VEGF-A rs699947 and / or ICAM-1 rs1799969 polymorphisms would benefit from adding bevacizumab to chemotherapy.

[0168] This discovery also allows physicians to administer bevacizumab in combination with fluoropyrimidine chemotherapy agents to treat mCRC based on the presence or absence of the SNPs VEGF-A rs699947 polymorphism and ICAM-1 rs1799969 polymorphism in a subject's genomic DNA.

[0169] The present invention provides methods for determining whether a subject having mCRC will respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or for predicting the overall survival (OS) of the subject, the methods comprising the steps of: (a) determining the presence or absence of the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism in a sample obtained from the subject, or identifying a subject identified as having the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism; and (b) indicating that the subject is likely to respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or to have a longer overall survival when at least one of the VEGF-A rs699947 polymorphism and ICAM-1 rs1799969 polymorphism is present. In such an example, the subject has the polymorphisms VEGF-A rs699947 polymorphism and ICAM-1 rs1799969 polymorphism.

[0170] For example, subjects with the VEGF-A rs699947 polymorphism treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 40 to 65 months, 45 to 60 months, or 50 to 55 months. For example, subjects with the VEGF-A rs699947 polymorphism treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 months, 34 months, 35 months, 36 months, 37 months, or 58 months. 3 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months of OS.

[0171] For example, subjects having the ICAM-1 rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months. For example, subjects having the ICAM-1 rs1799969 polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, or 63 months.

[0172] The present invention further demonstrates that if the VEGF-A rs699947 polymorphism and the ICAM-1 rs1799969 polymorphism are absent in a subject's sample, the subject is less likely to respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or less likely to have a prolonged overall survival. In such an example, the subject has wild-type genotypes of VEGF-A rs699947 (allele C / C) and ICAM-1 rs1799969 (allele G / G).

[0173] Identification of the polymorphism The sample is obtained from a subject diagnosed with mCRC (i.e., metastatic cancer of the colon or rectum). In a preferred embodiment, the subject is an adult.

[0174] The sample may be whole blood, serum, peripheral blood leukocytes, or saliva. Other samples from which genomic DNA can be extracted are known to those skilled in the art.

[0175] The presence or absence of the allele can be determined by standard methods known in the art for detecting polymorphisms, such as PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation signaling.

[0176] A common method for identifying polymorphisms is to (a) collecting a whole blood sample; (b) separating the blood and isolating a serum sample; (c) extracting genomic DNA from blood leukocytes; (d) measuring DNA concentration; (e) analyzing genomic variants by PCR and purifying the PCR products; (f) Perform our DNA sequence analysis on the purified PCR products.

[0177] In an alternative embodiment, a method for identifying the polymorphisms is to add a label that binds to the VEGF-A rs699947 polymorphism and / or a label that binds to the ICAM-1 rs1799969 polymorphism and detect one or both of the bound labels.

[0178] In an alternative embodiment, the method for identifying the polymorphisms is to add a label that binds to said VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism and detect the bound label.

[0179] In some embodiments of the invention, the subject has already been identified as having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism.

[0180] Bevacizumab and fluoropyrimidine chemotherapy agents The bevacizumab and fluoropyrimidine chemotherapy treatment can be a first-line therapy for mCRC. The bevacizumab and fluoropyrimidine chemotherapy can be administered as part of a standard chemotherapy regimen known in the art.

[0181] The fluoropyrimidine chemotherapy agent may be selected from the group consisting of 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI), 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX), capecitabine / irinotecan (BEV-CapIRI), capecitabine / oxaliplatin (BEV-CapOX), or 5-fluorouracil / leucovorin / oxaliplatin / irinotecan (FOLFOXIRI). Each fluoropyrimidine chemotherapy agent option in combination with bevacizumab is considered clinically equivalent to the other options in both the United States (NCCN) and Europe (ESMO). See Cutsem et al., Ann Oncol. 2016 Aug 27(8):1386-422 (https: / / www.ncbi.nlm.nih.gov / pubmed?term=27380959) and the NCCN Colorectal Cancer Guidelines (https: / / www.nccn.org / professionals / physician_gls / default.aspx).

[0182] The bevacizumab and fluoropyrimidine chemotherapeutic agent may be administered by any suitable mode of administration known to those skilled in the art. For example, the mode of administration may be parenteral, such as intramuscular, subcutaneous, and intravenous administration. This parenteral administration may be by bolus administration or by infusion over a period of time.

[0183] The bevacizumab may be administered at a dose of 2 to 12.5 mg / kg, 4 to 10 mg / kg, or 5 to 7.5 mg / kg of body weight, or at a dose of 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 5.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 7 mg / kg, 7.5 mg / kg, 8 mg / kg, 8.5 mg / kg, 9 mg / kg, 9.5 mg / kg, 10 mg / kg, 10.5 mg / kg, 11 mg / kg, 11.5 mg / kg, 12 mg / kg, or 12.5 mg / kg, and may be administered every week, every two weeks, every three weeks, or every four weeks. This treatment cycle may be repeated 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more times, i.e., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more times. There may be 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more treatment cycles of bevacizumab and a fluoropyrimidine chemotherapy agent.

[0184] In some embodiments, the bevacizumab is administered as an intravenous infusion at a dose of 5 mg / kg every two weeks in combination with the fluoropyrimidine chemotherapeutic agent. In some embodiments, the bevacizumab is administered as an intravenous infusion at a dose of 5 mg / kg every two weeks in combination with BEV-FOLFIRI or BEV-FOLFOX.

[0185] In some embodiments, the bevacizumab is administered as an intravenous infusion in combination with the fluoropyrimidine chemotherapeutic agent at a dose of 7.5 mg / kg every three weeks in a three-week cycle. It will be administered as an intravenous infusion in combination with BEV-CapIRI or BEV-CapOX in a 3-weekly cycle at a dose of 1000 mg / kg.

[0186] Intravenous infusion of bevacizumab or a bevacizumab biosimilar at a dose of 7.5 mg / kg every 3 weeks is considered clinically equivalent to intravenous infusion of 5 mg / kg every 2 weeks in combination with the fluoropyrimidine chemotherapy agent.

[0187] Intravenous administration of bevacizumab and fluoropyrimidine chemotherapy agents allows for rapid infusion into the systemic circulation, where the intravenously administered bevacizumab and fluoropyrimidine chemotherapy agents are transported throughout the body via the bloodstream. Intravenous administration allows for fast absorption times and flexibility in drug administration. Administration can be performed as an intravenous infusion over minutes to hours. Specifically, administration can be performed as an intravenous infusion over 30 to 90 minutes. A portable pump can be used to administer continuous, slow-flow infusion of bevacizumab and fluoropyrimidine chemotherapy agents, allowing for ongoing intravenous infusion.

[0188] The first-line therapy may be followed by maintenance therapy, which may be selected from the list of bevacizumab-mFOLFOX, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont (fluorouracil in combination with leucovorin or calcium folinate), bevacizumab-capecitabine, and / or bevacizumab monotherapy.

[0189] For example, after 6 cycles of FOLFOX / FOLFIRI or 4 cycles of CapeOx / CapIri, patients may continue on maintenance therapy based on response. The choice of this maintenance therapy needs to be individualized and depends on the following factors: 1. Tolerability and side effects 2. Socio-economic and logistical issues 3. Extent of disease, histology, grade, number of metastases, etc. Take into consideration:

[0190] For the most part, there is no clear evidence for the best treatment duration and regimen. 1. Bevacizumab monotherapy 2. Bevacizumab in combination with fluoropyrimidine alone 3. Continuation of initial bevacizumab and fluoropyrimidine chemotherapy and the fluoropyrimidine chemotherapy agent is 5-fluorouracil / leucovorin / irinotecan (BEV-FOLFIRI), 5-fluorouracil / leucovorin / oxaliplatin (BEV-FOLFOX), capecitabine / irinotecan (BEV-CapIRI), capecitabine / oxaliplatin (BEV-CapOX), or 5-fluorouracil / leucovorin / oxaliplatin / irinotecan (FOLFOXIRI).

[0191] For example, a subject who receives initial treatment consisting of two weeks of bevacizumab and a fluoropyrimidine chemotherapy agent may receive bevacizumab (7.5 mg / kg) every three weeks as maintenance treatment. As part of this maintenance dose, the bevacizumab may be administered at a dose of 2 to 12.5 mg / kg, 4 to 10 mg / kg, or 5 to 7.5 mg / kg of body weight, or at a dose of 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 5.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 7 mg / kg, 7.5 mg / kg, 8 mg / kg, 8.5 mg / kg, 9 mg / kg, 9.5 mg / kg, 10 mg / kg, 10.5 mg / kg, 11 mg / kg, 11.5 mg / kg, 12 mg / kg, or 12.5 mg / kg, and may be administered every week, every two weeks, every three weeks, or every four weeks. This maintenance treatment cycle may be repeated 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more times, i.e., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more times. There may be 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more maintenance treatment cycles of bevacizumab alone, with a fluoropyrimidine alone, or with a fluoropyrimidine-based chemotherapy agent.

[0192] The present invention provides methods for selecting a therapy for mCRC in a subject, the methods comprising: (a) determining the presence or absence of the VEGF-A rs699947 polymorphism (allele A / A) and / or the ICAM-1 rs1799969 polymorphism (allele G / A) in a sample obtained from the subject; and (b) selecting a therapy consisting of bevacizumab and a fluoropyrimidine chemotherapeutic agent when at least one of the VEGF-A rs699947 polymorphism (allele A / A) and the ICAM-1 rs1799969 polymorphism (allele G / A) is present.

[0193] The present invention further provides a method for selecting a treatment for mCRC in a subject, the method comprising the steps of (a) determining the presence or absence of the VEGF-A rs699947 polymorphism (allele A / A) and / or ICAM-1 rs1799969 polymorphism (allele G / A) in a sample obtained from the subject, and (b) selecting a treatment consisting of a fluoropyrimidine chemotherapeutic agent alone if the VEGF-A rs699947 polymorphism (allele A / A) and the ICAM-1 rs1799969 polymorphism (allele G / A) are absent in the subject's sample. In such an example, the subject has wild-type genotypes of VEGF-A rs699947 (allele C / C) and ICAM-1 rs1799969 (allele G / G).

[0194] The present invention provides a method of treating mCRC in a subject, comprising: (a) detecting the VEGF-A rs699947 polymorphism (allele A / A) and / or ICAM-1 in a sample obtained from the subject; The method further comprises the steps of (a) determining the presence or absence of the rs1799969 polymorphism (allele G / A); and (b) administering bevacizumab and a fluoropyrimidine chemotherapeutic agent if at least one of the VEGF-A rs699947 polymorphism (allele A / A) and the ICAM-1 rs1799969 polymorphism (allele G / A) is present.

[0195] The present invention provides a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent for use in a method for treating mCRC in a subject, the method comprising the steps of: (a) determining the presence or absence of the VEGF-A rs699947 polymorphism (allele A / A) and / or ICAM-1 rs1799969 polymorphism (allele G / A) in a sample obtained from the subject; and (b) administering bevacizumab and a fluoropyrimidine chemotherapeutic agent if at least one of the VEGF-A rs699947 polymorphism (allele A / A) and ICAM-1 rs1799969 polymorphism (allele G / A) is present.

[0196] The present invention also provides a method for treating mCRC in a subject having the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism, the method comprising administering bevacizumab and a fluoropyrimidine chemotherapeutic agent.

[0197] The present invention further includes a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent, said combination being used in a method for treating mCRC in a subject carrying the VEGF-A rs699947 polymorphism and / or the ICAM-1 rs1799969 polymorphism.

[0198] References to bevacizumab include references to Avastin® and apply equally to biosimilars of the reference drug / reference product bevacizumab (Avastin®) that have been granted marketing authorization by regulatory authorities such as the European Medicines Agency (EMA), the US Food and Drug Administration (FDA), etc. Accordingly, references to the combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent also apply to biosimilars of the reference drug / reference product bevacizumab (Avastin®) and a fluoropyrimidine chemotherapeutic agent.

[0199] Also provided as part of the present invention is a method of treating cancer in a subject with metastatic colorectal cancer (mCRC), comprising or consisting of the steps of administering to the subject bevacizumab and a fluoropyrimidine chemotherapeutic agent, and determining the presence or absence of the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism in a sample obtained from the subject, or identifying a subject identified as having the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism.

[0200] For subjects with the VEGF-A rs699947 polymorphism, when treated with bevacizumab and a fluoropyrimidine chemotherapy agent, the subject's outcome is an OS of 33 to 71 months, 33.3 to 70.7 months, 40 to 65 months, 45 to 60 months, or 50 to 55 months. For subjects with the VEGF-A rs699947 polymorphism, when treated with bevacizumab and a fluoropyrimidine chemotherapy agent, the subject's outcome is an OS of 33 months, 34 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months. When subjects with the ICAM-1 rs1799969 polymorphism are treated with bevacizumab and a fluoropyrimidine chemotherapy agent, the outcomes for those subjects are OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months. For subjects with the ICAM-1 rs1799969 polymorphism, when treated with bevacizumab and a fluoropyrimidine chemotherapy agent, the outcomes for the subjects are OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, and 63 months.

[0201] Also provided as part of the present invention is a method of treating metastatic colorectal cancer in a subject, the method comprising administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject known to have the ICAM-1 rs1799969 polymorphism.

[0202] Also provided as part of the present invention is a method for administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject with metastatic colorectal cancer, the method comprising identifying the presence or absence of the ICAM-1 rs1799969 polymorphism in a sample obtained from the subject, and administering the combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to the subject having the ICAM-1 rs1799969 polymorphism.

[0203] Also provided as part of the present invention is a method of treating metastatic colorectal cancer in a subject, the method comprising administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject known to have the VEGF-A rs699947 polymorphism.

[0204] Also provided as part of the present invention is a method for administering a combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to a subject with metastatic colorectal cancer, the method comprising identifying the presence or absence of the VEGF-A rs699947 polymorphism in a sample obtained from the subject, and administering the combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent to the subject having the VEGF-A rs699947 polymorphism.

[0205] In the methods described herein, the combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent is administered as a first line or second line therapy.

[0206] In the methods described herein, the identifying step comprises detecting the polymorphism by a method comprising one or more of PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next generation sequencing.

[0207] Bevacizumab and biosimilars specifically recognize and bind to vascular VEGF-A, inhibiting the formation of new blood vessel cells (angiogenesis), thereby treating mCRC. Biosimilars are approved according to the same standards of pharmaceutical quality, safety, and efficacy that apply to all biological drugs. Biosimilars are clinically equivalent to the reference drug / reference product, bevacizumab (Avastin®), without clinically meaningful differences. Therefore, subjects with the VEGF-A rs699947 polymorphism and / or ICAM-1 rs1799969 polymorphism respond similarly to treatments containing a biosimilar of the reference drug / reference product, bevacizumab (Avastin®), as they do to treatments containing the reference drug / reference product, bevacizumab (Avastin®).

[0208] Bevacizumab biosimilars are known in the art and examples include, but are not limited to, Mvasi (ABP215) or (bevacizumab-awwb) (Amgen) and Zirabev (Pfizer).

[0209] Mvasi (ABP215) is a recombinant IgG1 humanized monoclonal antibody produced in Chinese hamster ovary cells. Mvasi (ABP215) has the same primary structure as bevacizumab. Mvasi (ABP215) binds to vascular endothelial growth factor (VEGF), thereby inhibiting VEGF from binding to its receptors on the surface of endothelial cells. Neutralization of VEGF biological activity results in regression of tumor angiogenesis, normalizing remaining tumor angiogenesis, and inhibiting its formation.

[0210] Zirabev is a recombinant IgG1 humanized monoclonal antibody produced in Chinese hamster ovary cells. Zirabev binds to vascular endothelial growth factor (VEGF), thereby inhibiting VEGF binding to its receptors on the surface of endothelial cells. Neutralization of VEGF biological activity results in regression of tumor angiogenesis, normalization of remaining tumor angiogenesis, and inhibition of new tumor blood vessel formation, thereby suppressing tumor growth.

[0211] Examples of anti-cancer drugs include, but are not limited to, BCD-021 (Biocad, Russia), FKB238 (AstraZeneca / Kyowa Hakko Kirin Fujifilm Biologics, USA / Japan), BCD500 (BIOCND, Korea), Krabeva (Biocon, India), BI695502 (Boehringer Ingelheim, Germany), CT-P16 (Celltrion, Korea), CHS-5217 (Cohilus, USA), DRZ_BZ (Dr. Reddy's Laboratories, India), Cizumab (Hetero (Lupin), India), Bevax (mAbxience, Spain / Argentina), ONS-1045 (Oncobiolo A number of bevacizumab biosimilars are also in development, such as Zyx Pharmaceuticals / Viropro, USA), PF-06439535 (Pfizer, USA), HD204 (Prestige Biopharmaceuticals, Singapore), Bevacirel (Reliance Life Sciences / Lupin, India), and SB8 (Samsung Bioepis (Biogen / Samsung) / Merck, Korea / USA), which would be bevacizumab biosimilars in accordance with the present invention if the appropriate regulatory authorities, such as the European Medicines Agency (EMA) or the US Food and Drug Administration (FDA), approve these compounds as bevacizumab biosimilars.

[0212] Based on the fact that these biosimilars of the reference drug / product bevacizumab (Avastin®) share high structural and functional similarity to bevacizumab, the data provided herein can be extrapolated to biosimilars of the reference drug / product bevacizumab (Avastin®). Specifically, the biosimilars bind to human vascular endothelial growth factor (VEGF-A) and inhibit its biological activity. The interaction of VEGF with its receptors results in endothelial cell proliferation and angiogenesis in in vitro models of angiogenesis. The biosimilars bind to circulating VEGF-A and prevent it from binding to its receptors (VEGFR-1 and VEGFR-2) on the surface of endothelial cells, resulting in the inhibition of tumor angiogenesis, growth, and metastasis.

[0213] Biosimilars are highly similar to the reference drug / reference product, bevacizumab (Avastin®), in terms of structure, biological activity and efficacy, safety, and immunogenicity profile. These biosimilars must be clinically equivalent to the reference drug / reference product, bevacizumab (Avastin®), and there may be no clinically meaningful differences between the biosimilar and the reference drug / reference product, bevacizumab (Avastin®). Specifically, there may be no clinically meaningful differences between the biosimilar and bevacizumab (reference drug / reference product) in terms of safety, quality, and efficacy.

[0214] In some embodiments, the bevacizumab is the reference drug / product bevacizumab (Avastin®).

[0215] In some embodiments, the bevacizumab is a biosimilar to the reference drug / reference product bevacizumab (Avastin®). In some embodiments, the bevacizumab is a biosimilar to the reference drug / reference product bevacizumab (Avastin®) selected from the list consisting of Mvasi (ABP215) (Amgen) and Zirabev (Pfizer). These biosimilars are clinically equivalent to the reference drug / reference product bevacizumab (Avastin®) with no clinically meaningful differences.

[0216] In some embodiments, the bevacizumab may be approved by an appropriate regulatory authority, such as the European Medicines Agency (EMA), the U.S. Food and Drug Administration (FDA), as a biosimilar to the reference drug / product bevacizumab (Avastin®), in the form of BCD-021 (Biocad, Russia), FKB238 (AstraZeneca / Kyowa Hakko Kirin Fujifilm Biologics, U.S. / Japan), BCD500 (BIOCND, South Korea), Krabeva (Biocon, India), BI695502 (Boehringer Ingelheim, Germany), CT-P16 (Celltrion, South Korea), CHS-5217 (Cohilus, U.S.), DRZ_BZ (Dr. Ready's and biosimilars of bevacizumab (Avastin®), a reference drug / product selected from the list consisting of: Be ... [Example]

[0217] Patients and Treatment A confidential, real-world observational study was conducted at the Department of Oncology, Patras University Hospital, Greece, between July 2012 and July 2018. Patients were aged 18 years or older, had an ECOG activity status of 0-2, and had histopathologically confirmed mCRC. All patients received first-line treatment with bevacizumab in combination with fluoropyrimidine chemotherapy.

[0218] Bevacizumab (Avastin®) was administered intravenously at a dose of 5 mg / kg every 2 weeks in combination with 5-fluorouracil / leucovorin / irinotecan or oxaliplatin (BEV-FOLFIRI or BEV-FOLFOX, respectively) or at a dose of 7.5 mg / kg every 3 weeks in combination with capecitabine / irinotecan or oxaliplatin (BEV-CapIRI or BEV-CapOX, respectively) in 3-weekly cycles. Treatment was initially administered for 6 cycles (BEV-FOLFIRI, BEV-FOLFOX) or 4 cycles (BEV-CapIRI, BEV-CapOX), and responders continued bevacizumab-based maintenance therapy. Radiographic evaluations were performed every 8 to 12 weeks or as clinically indicated. Responses were assessed according to RECIST (Revised Edition).

[0219] Maintenance treatment was determined by the treating physician and was selected from bevacizumab-mFOLFOX, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, and bevacizumab monotherapy. Progression-free survival (PFS) and overall survival (OS) were the endpoints. The study was conducted in accordance with the Declaration of Helsinki and the International Council for Harmonisation (ICH) Good Clinical Practice. Approval was obtained from the Institutional Review Board. Signed informed consent was obtained from all patients before study enrollment.

[0220] Pharmacogenetic analysis Whole blood samples were collected at baseline for analysis of the VEGF-A SNPs (rs2010963, rs1570360, rs699947) and ICAM-1 SNPs (rs5498, rs1799969). Blood samples were collected in serum separator tubes and allowed to clot for 30 minutes. Serum was removed after centrifugation at 1000 × g for 20 minutes and stored in aliquots at −20°C or below until analysis. Genomic DNA was extracted from patients' peripheral blood leukocytes using the Gentra Puregene Blood Kit (QIAGEN) (see DNA Sequencing by Capillary Electrophoresis, Applied Biosystems Chemistry Guide, 2nd Edition, https: / / tools.thermofisher.com / content / sfs / manuals / cms_041003.pdf, incorporated herein by reference). DNA concentration was determined by measuring the optical density at 260 nm using a UV-visible spectrophotometer (NanoDrop 2000, Thermo Fisher Scientific). DNA purity, expressed as the ratio of optical densities at 260 nm to 280 nm, ranged from 1.7 to 1.9. Genomic variants of VEGF-α (rs699947, rs1570360, and rs2010963) and ICAM-1 (rs1799969, rs5498) were analyzed by polymerase chain reaction (PCR) according to the KAPA2G Fast HotStart protocol (Kapa Biosystems, Massachusetts, USA). PCR products were separated on a 1% (wt / vol) agarose gel, stained with Midori Green, and purified using a PCR fragment DNA purification kit (Dongsheng Biotech, DNA purity 1.7 to 1.9). After purification, samples were subjected to direct DNA sequence analysis on an ABI Prim 3130xl DNA Analyzer (Applied Biosystems) using the Big Dye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, CA, USA) according to the manufacturer's instructions.

[0221] The mutation status of KRAS, NRAS (exons 2, 3, and 4), and BRAF (exons 11 and 15) was determined in genomic DNA extracted from the patient's formalin-fixed, paraffin-embedded tissue samples using the QIAmp DNA FFPE Tissue Kit (Qiagen / MagCore Genomic DNA FFPE One-Step Kit, RBC Biosciences). Targeted realignment (Ion AmpliSeq Panel, Thermo Fisher Scientific) was used to detect mutations in exons 2, 3, and 4 of KRAS and NRAS, and exons 11 and 15 of BRAF. The next-generation sequencing platform Ion Proton (Thermo Fisher Scientific) was used for sequencing. The detection limit of this method was 2–5% (mutant / wild-type allele).

[0222] statistics All categorical data, including treatment modalities and polymorphisms, were tabulated and presented as frequencies and totals (see Table 1). Statistical significance was set at p<0.05.

[0223] We used Kaplan-Meier estimates for survival analysis. All losses to follow-up were censored at the most recent available time point. Median OS and PFS estimates are presented with 95% confidence intervals. The log-rank test was used to compare OS and PFS distributions for different polymorphisms, and the effects of age and sex were investigated in multiple analyses using Cox regression. Regarding sample size calculations, due to study availability limitations at the time of recruitment, we calculated the effect size that the study could capture for a given number of patients. With an 80% detection value and an α set at 0.05, we could detect at least a 1.5-year difference in median PFS and OS between different polymorphisms for 46 patients with a 60-month follow-up period and 1-year recruitment period. Kaplan-Meier graphs were generated using GraphPad Prism version 7 for Windows (GraphPad Software, La Jolla, CA, USA). Analyses were performed using SAS version 9.4, SAS Institute, Inc., Cary, North Carolina, 2015, SAS / IML® 14.1 User's Guide.

[0224] result Patient population A total of 46 patients with mCRC were enrolled in this study. Overall, patients had a mean age of 64.5 years (range, 31–86 years) and were predominantly male (28 / 46, 61%). The most commonly used initial treatment was BEV-FOLFOX (46%), and the most common maintenance treatment was bevacizumab monotherapy (30%) (Table 1). The mean number of metastatic sites was 2 (range, 1–5).

[0225] Genotype The majority of patients had wild-type variants of VEGF-A SNPs rs2010963 (58.7%), rs1570360 (67.4%), and rs699947 (74%). Similarly, wild-type ICAM-1 gene variant rs1799969 was predominant (78.3%), but 50% of patients were heterozygous for rs5498. Fifty percent of patients had tumors with wild-type KRAS, and 61.2% had tumors with NRAS mutations. The majority of patients (81.8%) had tumors with wild-type BRAF.

[0226] Genotype frequencies are summarized in Table 1.

[0227] Table 1: Baseline patient characteristics, treatment regimens used, and genotype frequencies [Table 2]

[0228] Table 1 (continued) [Table 3] Note: Bevacizumab-mFOLFOX6 (5 mg / kg bevacizumab, 85 mg / m² oxaliplatin, 400 mg / m² folinic acid, 400 mg / m² fluorouracil bolus, 2400 mg / m² fluorouracil over 46 hours every 2 weeks), bevacizumab-FOLFIRI (5 mg / kg bevacizumab, 180 mg / m² irinotecan, 400 mg / m² folinic acid, 400 mg / m² fluorouracil bolus, 2400 mg / m² fluorouracil over 46 hours every 2 weeks). bevacizumab-CapOX (7.5 mg / kg bevacizumab on day 1, 130 mg / m² oxaliplatin on day 1, and 1000 mg / m² capecitabine every 12 hours from days 1 to 14 every 3 weeks), bevacizumab-CapIRI (7.5 mg / kg bevacizumab on day 1, 250 mg / m² irinotecan on day 1, and 1000 mg / m² capecitabine every 12 hours from days 1 to 14 every 3 weeks), bevacizumab-De Gramont (5 mg / kg bevacizumab every 2 weeks, 200 mg / m² folinic acid on days 1 and 2, 400 mg / m² fluorouracil bolus on days 1 and 2, 2400 mg / m² fluorouracil over 22 hours on days 1 and 2), bevacizumab-capecitabine (7.5 mg / kg bevacizumab on day 1, 1000 mg / m² capecitabine every 12 hours on days 1-14 every 3 weeks), bevacizumab monotherapy (5 mg / kg every 2 weeks or 7.5 mg / kg every 3 weeks).

[0229] Clinical response After a median follow-up of 60 months, 88.1% of patients had progressive disease and 58.0% had died.

[0230] The median PFS and OS for all patients were 9 and 18 months, respectively. Regarding response, 1 patient achieved a complete response (CR), 34.8% of patients achieved a partial response (PR), 54.4% of patients achieved stable disease, and 8.7% of patients achieved progressive disease (PD), all at the time of first imaging evaluation.

[0231] Table 2. Polymorphism analysis of response assessment by RECIST 1.1 and DCR [Table 4]

[0232] Table 2 (continued) [Table 5] Note: DCR: disease control rate, CR: complete response, PR: partial response, SD: stable disease, PD: progressive disease.

[0233] The impact of genetic polymorphisms on treatment efficacy The presence of any polymorphisms in the VEGF-A, ICAM-1, KRAS, and NRAS genes did not affect DCR (Table 2). In contrast, BRAF mutation status was significantly (p = 0.026) associated with resistance to treatment, as 50% of patients with BRAF-mutant tumors had progressive disease (PD) at the time of first imaging evaluation. On the other hand, disease control was achieved in 100% of patients with tumors harboring wild-type BRAF.

[0234] Patients homozygous for VEGF-A rs699947A / A had a significantly (p=0.006) prolonged PFS of 32.6 months compared with 8.1 months in patients with the wild-type C / C variant (Table 3 and Figure 3). VEGF-A rs1570360 and rs2010963, ICAM-1 rs5498 and rs1799969, KRAS gene variants, and NRA No other statistical significance was found between S gene variants and BRAF gene variants and PFS (Table 3).

[0235] Unexpectedly, VEGF-A In patients with rs699947A / A, OS was significantly prolonged to 59.4 months (p=0.043) (Table 3 and Figure 1). Therefore, patients with VEGF-A rs699947 A / A allele are significantly associated with prolonged OS and are more likely to respond to treatment with bevacizumab and fluoropyrimidine chemotherapy.

[0236] Patients heterozygous for VEGF-A rs1570360G / A had longer OS compared with carriers of the G / G or A / A variants (43.2, 27.8, and 39.2 months, respectively), although the difference did not reach statistical significance (p=0.074). None of the variants of VEGF-A rs2010963 had an association with OS (p=0.811).

[0237] Unexpectedly, patients with the ICAM-1 rs1799969 G / A allele had a median OS of 48.7 months compared with 29.1 months for patients with the G / G allele (p = 0.036) (Table 3 and Figure 2). Thus, patients with the ICAM-1 rs1799969 G / A allele were significantly associated with prolonged OS and were more likely to respond to treatment with bevacizumab and fluoropyrimidine-based chemotherapy.

[0238] ICAM-1 rs5498 was not significantly associated with OS (p=0.159). Also, the median OS of patients with wild-type BRAF was significantly longer than that of patients with mutant BRAF (16.7 months vs. 6.8 months, p=0.027). KRAS and NRAS were not significantly associated with OS (p-values ​​of 0.511 and 0.374, respectively, Table 3). Age and gender did not appear to have any statistically significant association with DCR, PFS, or OS in patients with mCRC treated with bevacizumab and fluoropyrimidine chemotherapy.

[0239] Table 3. Polymorphism analysis for PFS and OS [Table 6]

[0240] Table 3 (continued) [Table 7] Note: PFS: progression-free survival; OS: overall survival.

[0241] Consideration Our results demonstrate a statistically significant association between specific polymorphisms in VEGF-dependent and non-VEGF-dependent genes with improved clinical outcomes in patients with mCRC treated with bevacizumab and fluoropyrimidine chemotherapy. Patients homozygous for VEGF-A rs699947A / A had significantly higher rates of VEGF-A mutations compared with carriers of the wild-type C / C variant. Patients with mCRC who were treated with bevacizumab in combination with a fluoropyrimidine chemotherapy agent had significantly prolonged PFS and OS (32.6 vs. 8.1 months PFS and 59.4 vs. 16.9 months OS, respectively). This is the first time that a statistically significant association has been observed between VEGF-A rs699947A / A and OS in patients with mCRC treated with bevacizumab in combination with a fluoropyrimidine chemotherapy agent. This provides a validated biomarker that can optimize the management of mCRC by selecting appropriate treatment. One of the major benefits of this study is the 60-month (i.e., 5-year) follow-up period, which will provide better long-term survival data.

[0242] Additionally, carriers of VEGF-A rs1570360G / A and A / A variants tended to have longer OS, at 43.2 months and 39.2 months, respectively, compared with 27.8 months for the corresponding G / G variant.

[0243] It has been reported that no correlation was found between VEGF-A polymorphisms (including rs699947) and OS in patients with mCRC treated with bevacizumab (see Cui et al., Oncotarget, 2017, Vol. 8 (62), pp. 105472-104478, incorporated herein by reference). The discrepancy between our results and those of Cui et al. may be due to differences in study design and / or statistical analysis. Cui et al. also noted that their results were limited by a relatively small sample size and a single ethnic affiliation.

[0244] Genotyping of 173 mCRC patients treated with bevacizumab in combination with FOLFIRI or CapIRI showed that rs1570360G / G was significantly associated with decreased OS compared with G / A.

[0245] In our study, OS was significantly associated with the ICAM-1 rs1799969G / A variant compared with the G / G variant (48.7 months versus 29.1 months, respectively, p=0.036). This is the first time that a significant association has been observed between the ICAM-1 gene and OS in mCRC patients treated with bevacizumab in combination with fluoropyrimidine chemotherapy agents.

[0246] Our results show that mCRC carriers of the ICAM-1 rs1799969G / A allele have a 19.6-month survival benefit compared with carriers of the wild-type G / G allele when treated with bevacizumab in combination with fluoropyrimidine chemotherapy, providing a validated biomarker that can optimize the management of mCRC by selecting appropriate treatment.

[0247] It is also important to clarify that this study was a practical one, and therefore the chemotherapy combination was not uniform for all patients. This is the clinical treatment of patients in hospitals. Identifying highly significant SNPs may subsequently have an impact on delaying disease progression, thus increasing patient survival and quality of life.

[0248] In summary, we have identified polymorphisms in key angiogenesis and intracellular signaling pathways, VEGF-A rs699947A / A and ICAM-1 rs1799969G / A, as predictors of response to first-line bevacizumab and fluoropyrimidine chemotherapy in mCRC.

[0249] As used in this specification and claims, the terms "comprises" and "comprising" and their variations mean that the specified features, steps, or integers are included. The term is not to be interpreted to exclude the presence of other features, steps or components.

[0250] The features disclosed in the foregoing description or the following claims or accompanying drawings, presented in a particular form or in terms of means for performing the disclosed functions, or suitable methods or processes for obtaining the disclosed results, may be used alone or in any combination of such features to realize the invention in its various forms.

[0251] While certain example embodiments of the present invention have been described, the appended claims are not intended to be limited solely to these embodiments. The claims should be interpreted literally, purportedly, and / or encompass equivalents.

Claims

1. 1. A method for obtaining data for determining whether a subject with metastatic colorectal cancer (mCRC) will respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or for predicting overall survival (OS) of said subject, comprising: (a) determining the presence or absence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism in a sample obtained from the subject; and (b) indicating that the subject is likely to respond to treatment with bevacizumab and a fluoropyrimidine chemotherapeutic agent and / or is likely to have a longer overall survival if treated with bevacizumab and a fluoropyrimidine chemotherapeutic agent if at least one of the ICAM-1 rs1799969G / A allele polymorphism and the VEGF-A rs699947A / A allele polymorphism is present. comprising or consisting of the steps of The fluoropyrimidine chemotherapeutic agent is (1) 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI); (2) 5-fluorouracil, leucovorin, and oxaliplatin (FOLFOX); (3) capecitabine and irinotecan (CapIRI); and (4) Capecitabine and oxaliplatin (CapOX) The method is selected from the group consisting of:

2. 1. A method for obtaining data for selecting a treatment for metastatic colorectal cancer in a subject, comprising: (a) determining the presence or absence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism in a sample obtained from the subject; and (b) obtaining data for selecting the treatment method comprising or consisting of bevacizumab and a fluoropyrimidine chemotherapeutic agent when at least one of the ICAM-1 rs1799969G / A allele polymorphism and the VEGF-A rs699947A / A allele polymorphism is present; comprising or consisting of the steps of The fluoropyrimidine chemotherapeutic agent is (1) 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI); (2) 5-fluorouracil, leucovorin, and oxaliplatin (FOLFOX); (3) capecitabine and irinotecan (CapIRI); and (4) Capecitabine and oxaliplatin (CapOX) The method is selected from the group consisting of:

3. A combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent, (a) determining the presence or absence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism in a sample obtained from a subject, or identifying a subject identified as having the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism; and (b) administering bevacizumab and a fluoropyrimidine chemotherapeutic agent when at least one of the ICAM-1 rs1799969G / A allele polymorphism and the VEGF-A rs699947A / A allele polymorphism is present. wherein the method comprises administering to the subject a therapeutically effective amount of a compound selected from the group consisting of: The fluoropyrimidine chemotherapeutic agent is (1) 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI); (2) 5-fluorouracil, leucovorin, and oxaliplatin (FOLFOX); (3) capecitabine and irinotecan (CapIRI); and (4) Capecitabine and oxaliplatin (CapOX) The combination is selected from the group consisting of:

4. 1. A combination of bevacizumab and a fluoropyrimidine chemotherapeutic agent for use in a method for treating metastatic colorectal cancer in a subject with an ICAM-1 rs1799969G / A allele polymorphism and / or a VEGF-A rs699947A / A allele polymorphism, comprising: The fluoropyrimidine chemotherapeutic agent is (1) 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI); (2) 5-fluorouracil, leucovorin, and oxaliplatin (FOLFOX); (3) capecitabine and irinotecan (CapIRI); and (4) Capecitabine and oxaliplatin (CapOX) The combination is selected from the group consisting of:

5. 3. The method of claim 1 or 2, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

6. The method of any one of claims 1 to 2 and 5, wherein the presence or absence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism is identified by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next-generation sequencing.

7. The method according to any one of claims 1 to 2 and 5 to 6, wherein the treatment using bevacizumab and a fluoropyrimidine chemotherapeutic agent is a treatment in which the bevacizumab is administered as an intravenous infusion in combination with the fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg once every two weeks.

8. 7. The method according to any one of claims 1 to 2 and 5 to 6, wherein the treatment using bevacizumab and a fluoropyrimidine chemotherapeutic agent is a treatment in which the bevacizumab is administered as an intravenous infusion in combination with the fluoropyrimidine chemotherapeutic agent at a dose of 5 to 10 mg / kg once every three weeks in a cycle of every three weeks.

9. 9. The method of any one of claims 1 to 2 and 5 to 8, wherein the treatment is administered for 1 cycle, 2 cycles, 3 cycles, 4 cycles, 5 cycles, 6 cycles, 7 cycles, 8 cycles, 9 cycles, 10 cycles, 11 cycles, 12 cycles, 13 cycles, 14 cycles, 15 cycles, 16 cycles, 17 cycles, 18 cycles, 19 cycles, 20 cycles, 21 cycles, 22 cycles, 23 cycles, 24 cycles, 25 cycles, 26 cycles, 27 cycles, 28 cycles, 29 cycles, or 30 cycles.

10. 10. The method of any one of claims 1 to 2 and 5 to 9, wherein the presence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism in a sample obtained from the subject indicates that the subject is also likely to respond to maintenance treatment and / or may also have a longer overall survival with maintenance treatment, The method, wherein the maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, or bevacizumab monotherapy.

11. 11. The method of claim 10, wherein the maintenance treatment is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 cycles.

12. subjects having the ICAM-1 rs1799969G / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months, or 12. The method of any one of claims 1-2 and 5-11, wherein a subject having the rs1799969G / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, or 63 months.

13. subjects having the VEGF-A rs699947A / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 40 to 65 months, 45 to 60 months, or 50 to 55 months; or 13. The method of any one of claims 1-2 and 5-12, wherein a subject having the rs699947A / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 34 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

14. The combination of claim 3, wherein the sample is whole blood, serum, peripheral blood leukocytes, or saliva.

15. The combination according to any one of claims 3 to 4 and 14, wherein the presence or absence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism is identified by PCR, PCR-RFLP, direct sequencing, TaqMan, and / or next-generation sequencing.

16. The combination according to any one of claims 3 to 4 and 14 to 15, wherein the treatment using bevacizumab and a fluoropyrimidine chemotherapeutic agent is a treatment in which the bevacizumab is administered as an intravenous infusion in combination with the fluoropyrimidine chemotherapeutic agent at a dose of 2.5 to 7.5 mg / kg once every two weeks.

17. 16. The combination according to any one of claims 3 to 4 and 14 to 15, wherein the treatment using bevacizumab and a fluoropyrimidine chemotherapeutic agent is a treatment in which the bevacizumab is administered as an intravenous infusion in combination with the fluoropyrimidine chemotherapeutic agent in a cycle of every 3 weeks at a dose of 5 to 10 mg / kg once every 3 weeks.

18. 18. The combination according to any one of claims 3 to 4 and 14 to 17, wherein the treatment is administered for 1 cycle, 2 cycles, 3 cycles, 4 cycles, 5 cycles, 6 cycles, 7 cycles, 8 cycles, 9 cycles, 10 cycles, 11 cycles, 12 cycles, 13 cycles, 14 cycles, 15 cycles, 16 cycles, 17 cycles, 18 cycles, 19 cycles, 20 cycles, 21 cycles, 22 cycles, 23 cycles, 24 cycles, 25 cycles, 26 cycles, 27 cycles, 28 cycles, 29 cycles, or 30 cycles.

19. 19. A combination according to any one of claims 3 to 4 and 14 to 18, wherein the presence of the ICAM-1 rs1799969G / A allele polymorphism and / or the VEGF-A rs699947A / A allele polymorphism in a sample obtained from the subject indicates that the subject is also likely to respond to maintenance treatment and / or may also have a longer overall survival with maintenance treatment, A combination wherein said maintenance treatment is bevacizumab-mFOLFOX6, bevacizumab-FOLFIRI, bevacizumab-CapIRI, bevacizumab-De Gramont, bevacizumab-capecitabine, or bevacizumab monotherapy.

20. 20. The combination of claim 19, wherein the maintenance treatment is maintenance treatment administered for 1 cycle, 2 cycles, 3 cycles, 4 cycles, 5 cycles, 6 cycles, 7 cycles, 8 cycles, 9 cycles, 10 cycles, 11 cycles, 12 cycles, 13 cycles, 14 cycles, 15 cycles, 16 cycles, 17 cycles, 18 cycles, 19 cycles, 20 cycles, 21 cycles, 22 cycles, 23 cycles, 24 cycles, 25 cycles, 26 cycles, 27 cycles, 28 cycles, 29 cycles, or 30 cycles.

21. subjects having the ICAM-1 rs1799969G / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 34 to 63 months, 34.5 to 62.8 months, 40 to 60 months, 45 to 55 months, or 50 to 55 months, or 21. The combination of claims 3-4 and 14-20, wherein a subject having the rs1799969G / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 34 months, 34 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, or 63 months.

22. subjects having the VEGF-A rs699947A / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent have an OS of 33 to 71 months, 33.3 to 70.7 months, 40 to 65 months, 45 to 60 months, or 50 to 55 months; or 22. The combination of claims 3-4 and 14-21, wherein a subject having the rs699947A / A allele polymorphism when treated with bevacizumab and a fluoropyrimidine chemotherapy agent will have an OS of 33 months, 34 months, 35 months, 36 months, 37 months, 38 months, 39 months, 40 months, 41 months, 42 months, 43 months, 44 months, 45 months, 46 months, 47 months, 48 ​​months, 49 months, 50 months, 51 months, 52 months, 53 months, 54 months, 55 months, 56 months, 57 months, 58 months, 59 months, 60 months, 61 months, 62 months, 63 months, 67 months, 68 months, 69 months, 70 months, or 71 months.

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