Treatment of glioblastoma with desmopressin

Abstract

The application is directed towards methods of treating or preventing glioblastoma by administration of desmopressin, or other vasopressin analogues, either alone or in combination with temozolomide. Methods of treatment wherein the level of the arginine vasopressin type 2 receptor (AVPR2) polypeptide is measured in a subject having glioblastoma, and desmopressin is administered if said level is high, are also envisioned.

Description

TREATMENT OF GLIOBLASTOMA WITH DESMOPRESSINFIELD

[0001] The present disclosure relates generally to the treatment of glioblastoma.BACKGROUND

[0002] Glioblastoma (GBM) is an aggressive and complex form of brain cancer, characterized by poor overall survival rate. The median survival for patients diagnosed with GBM typically ranges from 12 to 18 months, with less than 5% of patients surviving beyond five years despite aggressive treatment strategies that include surgery, chemotherapy, and radiotherapy (Mohammed et al.).

[0003] The tumour microenvironment of glioblastoma exhibits significant plasticity, which enhances the tumour's resistance to standard therapies and challenges the effectiveness of treatment outcomes. (Sharma et al.).

[0004] The combination of surgical intervention followed by adjuvant therapies based on the use of Temozolomide, an alkylating agent, has resulted in only marginal improvements in survival rates, with many patients experiencing recurrence shortly after initial treatment (Alves et al).

[0005] T umour recurrence following surgical resection of glioblastoma is a significant challenge, primarily due to the presence of residual malignant cells that can survive and proliferate outside the resected tumour area (Celiku et al.). GBM cells possess an ability to infiltrate the brain parenchyma, demonstrating both high migratory capacity and intrinsic plasticity that allows them to adapt to various microenvironmental pressures. GBM cells can migrate as single entities, employing a mesenchymal mode of invasion characterized by saltatory movement, which enhances their ability to evade therapeutic interventions (Vollmann et al.). This adaptability not only aids in their local infiltration but also enables them to colonize distant sites within the brain, contributing to tumour recurrence and challenging treatment outcomes. Additionally, recurrent tumours frequently retain the genetic and epigenetic characteristics of their primary counterparts, yet they also exhibit new mutations and altered signaling pathways that contribute to their malignant behavior. Tumour-associated angiogenesis plays a crucial role in GBM progression, as the interaction between tumour cells and the tumour immune microenvironment (TiME) significantly influences vascular development and tumour growth. This dynamic interplay between angiogenesis and the TiME is involved in maintaining the tumour's metabolic demands and facilitating its invasive behavior,ultimately contributing to therapeutic resistance and poor patient outcomes.Consequently, this phenotypic plasticity of GBM cells not only supports tumour regrowth but also poses significant challenges for effective therapeutic intervention, necessitating the exploration of novel treatment strategies (Birzu et al.).

[0006] Another aspect is cerebral edema, which is a factor contributing to morbidity and mortality in several forms of brain injury, including GBM. It is necessary to consider the pathobiological phenomena underlying cerebral edema during surgery and chemoradiation treatment of brain tumours in order to identify strategies for its management.

[0007] There remains a need for a treatment of glioblastoma.SUMMARY

[0008] In one or more embodiments of the present disclosure, there is provided:

[0009] 1. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising, or consisting of: administering a therapeutically effective amount of desmopressin.

[0010] 2. The method of embodiment 1, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

[0011] 3. The method of embodiment 1 or 2, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0012] 4. The method of any one of embodiments 1 to 3, wherein desmopressin is administered before, during, and / or after surgery to remove the glioblastoma.

[0013] 5. The method of any one of embodiments 1 to 4, wherein desmopressin is administered perioperatively.

[0014] 6. The method of any one of embodiments 1 to 5, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0015] 7. The method of any one of embodiments 1 to 6, further comprising administering a therapeutically effective amount of temozolomide.

[0016] 8. The method of embodiment 7, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

[0017] 9. The method of any one of embodiments 7 to 8, wherein desmopressin is first administered to the subject followed by temozolomide administration to the subject.

[0018] 10. The method of embodiment 9, wherein temozolomide is administered from about 3 days to about 7 days following desmopressin administration, from about 2 days to about 10 days following desmopressin administration, from about 1 day to about 12 days following desmopressin administration, from about 30 minutes to about 60 minutes following desmopressin administration.

[0019] 11. The method of any one of embodiments 1 - 10, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0020] 12. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising, or consisting of:

[0021] - administering desmopressin prior to surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery;

[0022] - administering desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma.

[0023] 13 The method of embodiment 12, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg

[0024] 14. The method of embodiment 12 or 13, further comprising administering temozolomide to said subject.

[0025] 15. The method of embodiment 14, where said temozolomide is administered at a dose of about 75 mg / m2 daily for about 42 days concomitant with radiotherapy.

[0026] 16. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising or consisting of: administering a therapeutically effective amount of a vasopressin analogue.

[0027] 17 The method of any one of embodiment 16, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

[0028] 18. The method of embodiment 16 or 17 wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0029] 19. The method of any one of embodiment 16 to 18, wherein vasopressin analogue is administered before, during, and / or after surgery to remove the glioblastoma.

[0030] 20. The method of any one of embodiments 16 to 19, wherein the vasopressin analogue is administered perioperatively.

[0031] 21. The method of any one of embodiments 16 to 20, wherein the vasopressin analogue is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0032] 22. The method of any one of embodiments 16 to 21 , further comprising administering a therapeutically effective amount of temozolomide.

[0033] 23. The method of embodiment 22, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2 daily.

[0034] 24. The method of any one of embodiments 22 to 23, wherein the vasopressin analogue is first administered to the subject followed by temozolomide administration to the subject.

[0035] 25. The method of any one of embodiments 22 to 24, wherein temozolomide is administered from about 3 days to about 7 days following the vasopressin analogue administration, from about 2 days to about 10 days following the vasopressin analogue administration, from about 1 days to about 12 days following the vasopressin analogue administration, from about 30 minutes to about 60 minutes following the vasopressin analogue administration.

[0036] 26. The method of any one of embodiments 16 - 25, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0037] 27. The method of any one of embodiments 16 to 26, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0038] 28. The method of any one of embodiments 1 to 27, wherein the subject is a human.

[0039] 29. A method for treating or preventing glioblastoma in a subject, comprising or consisting of: measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma ;

[0040] and administering to a subject in need thereof a therapeutically effective amount of desmopressin, when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

[0041] 30. The method of embodiment 29, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

[0042] 31. The method of embodiment 30, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0043] 32. The method of any one of embodiments 29 or 31, wherein desmopressin is administered before, during, and / or after surgery to remove the glioblastoma.

[0044] 33. The method of any one of embodiments 29 to 32, wherein desmopressin is administered perioperatively.

[0045] 34. The method of any one of embodiments 29 to 33, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0046] 35. The method of any one of embodiments 29 to 34, further comprising administering a therapeutically effective amount of temozolomide.

[0047] 36. The method of embodiment 35, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2 daily.

[0048] 37. The method of any one of embodiments 35 to 36, wherein desmopressin is first administered to the subject followed by temozolomide administration to the subject.

[0049] 38. The method of any one of embodiments 35 to 37, wherein temozolomide is administered from about 3 days to about 7 days following desmopressin administration, from about 2 days to about 10 days following desmopressin administration, from about 1 days to about 12 days following desmopressin administration, from about 30 minutes to about 60 minutes following desmopressin administration.

[0050] 39. The method of any one of embodiments 29 to 38, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0051] 40. The method of embodiment 29, further comprising:

[0052] - administering desmopressin prior to surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery; and

[0053] - administering desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma.

[0054] 41. The method of embodiment 40, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg.

[0055] 42. The method of embodiment 40 or 41 , further comprising administering temozolomide to said subject.

[0056] 43. The method of embodiment 42, where said temozolomide is administered at a dose of about 75 mg / m2 daily for about 42 days concomitant with radiotherapy.

[0057] 44. The method of any one of embodiment 29 to 43, wherein said subject is a human.

[0058] 45. A method for treating or preventing glioblastoma in a subject, comprising, or consisting of, measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject;

[0059] and administering to a subject in need thereof a therapeutically effective amount of a vasopressin analogue, optionally in a physiological carrier or a pharmaceutically acceptable salt thereof, when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

[0060] 46. The method of embodiment 45, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0061] 47. The method of embodiment 45 or 46, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

[0062] 48. The method of any one of embodiments 45 to 47, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

[0063] 49. The method of any one of embodiments 45 to 48, wherein vasopressin analog is administered before, during, and / or after surgery to remove the glioblastoma.

[0064] 50. The method of any one of embodiments 45 to 49, wherein vasopressin analog is administered perioperatively.

[0065] 51. The method of any one of embodiments 45 to 50, wherein the vasopressin analog is administered at a dose of between about 0.1 pg / kg to about 1.0pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0066] 52. The method of any one of embodiments 45 to 51 , further comprising administering a therapeutically effective amount of temozolomide.

[0067] 53. The method of embodiment 52, wherein temozolomide administered at a dose of from 75 to 200 mg / m2 daily.

[0068] 54. The method of any one of embodiments 45 to 53, wherein vasopressin analog is first administered to the subject followed by temozolomide administration.

[0069] 55. The method of any one of embodiments 45 to 54, wherein temozolomide is administered from about 3 days to about 7 days following desmopressin administration, from about 2 days to about 10 days following desmopressin administration, from about 1 days to about 12 days following desmopressin administration, from about 30 minutes to about 60 minutes following desmopressin administration.

[0070] 56. The method of any one of embodiments 45 to 55, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0071] 57. The method of any one of embodiments 45 - 56, wherein the subject is a human.

[0072] 58. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising ,or consisting of:

[0073] - administering desmopressin prior to the surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery;

[0074] - administering desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma; and

[0075] - administration to said subject temozolomide at a dose of about 75 mg / m2 daily for about 42 days concomitant with radiotherapy;

[0076] - administering temozolomide for 6 cycles of maintenance treatment; and

[0077] - administering desmopressin on day 1 of each said cycle.

[0078] 59. The method of embodiment 58, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

[0079] 60. The method of embodiment 58 or 59, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0080] 61. The method of any one of embodiments 58 to 60, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0081] 62. The method of any one of embodiments 58 to 61, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

[0082] 63. The method of any one of embodiments 58 to 62, wherein said subject is a human.

[0083] 64. A kit for treating glioblastoma in a human, comprising or consisting of: desmopressin and TMZ, and optionally a container and optionally instructions for the use thereof.

[0084] 65. Use of a therapeutically effective amount of desmopressin for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma.

[0085] 66. The use of embodiment 65, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

[0086] 67. The use of embodiment 65 or 66, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0087] 68. The use of any one of embodiments 65 to 67, wherein desmopressin is for administration before, during, and / or after surgery to remove the glioblastoma.

[0088] 69. The use of any one of embodiments 65 to 68, wherein desmopressin is for administration perioperatively.

[0089] 70. The use of any one of embodiments 65 to 69, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0090] 71. The use of any one of embodiments 65 to 70, further comprising use of a therapeutically effective amount of temozolomide.

[0091] 72. The use of embodiment 71, wherein temozolomide is for administration at a dose of from 75 to 200 mg / m2daily.

[0092] 73. The use of any one of embodiments 71 to 72, wherein desmopressin is for administration first followed by temozolomide administration.

[0093] 74. The use of embodiment 73, wherein temozolomide is for administration from about 3 days to about 7 days following use of desmopressin, from about 2 days to about 10 days following use of desmopressin, from about 1 day to about 12 days following use of desmopressin, from about 30 minutes to about 60 minutes following use of desmopressin.

[0094] 75. The use of any one of embodiments 65 - 74, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0095] 76. Use of desmopressin prior to surgery to remove some or all of a glioblastoma, preferably about 30 minutes to about 60 before surgery; and

[0096] - administering use of desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma, for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma.

[0097] 77 The use of embodiment 76, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg.

[0098] 78. The use of embodiment 76 or 77, further comprising use of temozolomide to said subject.

[0099] 79. The use of embodiment 78, where said temozolomide is for administration at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy.

[0100] 80. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising or consisting of: administering Use of a therapeutically effective amount of a vasopressin analogue for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma.

[0101] 81 The use of any one of embodiment 80, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

[0102] 82. The use of embodiment 80 or 81 wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0103] 83. The use of any one of embodiment 80 to 82, wherein vasopressin analogue is for administration before, during, and / or after surgery to remove the glioblastoma.

[0104] 84. The use of any one of embodiments 80 to 83, wherein the vasopressin analogue is for administration perioperatively.

[0105] 85. The use of any one of embodiments 80 to 84, wherein the vasopressin analogue is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0106] 86. The use of any one of embodiments 80 to 85, further comprising use of a therapeutically effective amount of temozolomide.

[0107] 87. The use of embodiment 86, wherein temozolomide is for administration at a dose of from 75 to 200 mg / m2daily.

[0108] 88. The use of any one of embodiments 86 to 87, wherein the vasopressin analogue is for administration first followed by use of temozolomide.

[0109] 89. The use of any one of embodiments 86 to 88, wherein temozolomide is for administration from about 3 days to about 7 days following use of the vasopressin analogue, from about 2 days to about 10 days following use of the vasopressin analogue, from about 1 days to about 12 days following use of the vasopressin analogue, from about 30 minutes to about 60 minutes following use of the vasopressin analogue.

[0110] 90. The use of any one of embodiments 80 - 89, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0111] 91. The use of any one of embodiments 80 to 90, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0112] 92. The use of any one of embodiments 65 to 91, wherein the subject is a human.

[0113] 93. Use of a therapeutically effective amount of desmopressin for treating or preventing glioblastoma in a subject, or in the manufacture of a medicament for treating or preventing glioblastoma in a subject, comprising measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, wherein the use of desmopressin is indicated whenthe amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

[0114] 94. The use of embodiment 93, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

[0115] 95. The use of embodiment 94, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0116] 96. The use of any one of embodiments 93 or 95, wherein desmopressin is for administration before, during, and / or after surgery to remove the glioblastoma.

[0117] 97. The use of any one of embodiments 93 to 96, wherein desmopressin is for administration perioperatively.

[0118] 98. The use of any one of embodiments 93 to 97, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0119] 99. The use of any one of embodiments 93 to 98, further comprising use of a therapeutically effective amount of temozolomide.

[0120] 100. The use of embodiment 99, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

[0121] 101. The use of any one of embodiments 99 to 100, wherein desmopressin is for administration first followed by use of temozolomide.

[0122] 102. The use of any one of embodiments 99 to 101, wherein temozolomide is for administration from about 3 days to about 7 days following use of desmopressin, from about 2 days to about 10 days following use of desmopressin, from about 1 days to about 12 days following use of desmopressin, from about 30 minutes to about 60 minutes following use of desmopressin.

[0123] 103. The use of any one of embodiments 93 to 102, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0124] 104. The use of embodiment 93, further comprising:

[0125] - use of desmopressin prior to surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery; and

[0126] - use of desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma.

[0127] 105. The use of embodiment 104, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg.

[0128] 106. The use of embodiment 104 or 105, further comprising use of temozolomide.

[0129] 107. The use of embodiment 106, where said temozolomide is for administration at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy.

[0130] 108. The use of any one of embodiment 93 to 107, wherein said subject is a human.

[0131] 109. Use of a therapeutically effective amount of of a vasopressin analogue for treating or preventing glioblastoma in a subject, or in the manufacture of a medicament for treating or preventing glioblastoma in a subject, comprising measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, wherein the use of ther vasopressin analogue is indicated when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

[0132] 110. The use of embodiment 109, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0133] 111. The use of embodiment 109 or 110, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

[0134] 112. The use of any one of embodiments 109 to 111, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

[0135] 113. The use of any one of embodiments 109 to 112, wherein vasopressin analog is for administration before, during, and / or after surgery to remove the glioblastoma.

[0136] 114. The use of any one of embodiments 109 to 113, wherein vasopressin analog is for administration perioperatively.

[0137] 115. The use of any one of embodiments 109 to 114, wherein the vasopressin analog is for administration at a dose of between about 0.1 pg / kg to about1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0138] 116. The use of any one of embodiments 109 to 117, further comprising use of a therapeutically effective amount of temozolomide.

[0139] 117. The use of embodiment 116, wherein temozolomide for administration at a dose of from 75 to 200 mg / m2daily.

[0140] 118. The use of any one of embodiments 109 to 117, wherein vasopressin analog is for administration first followed by use of temozolomide administration.

[0141] 119. The use of any one of embodiments 109 to 118, wherein temozolomide is for administration from about 3 days to about 7 days following use of desmopressin, from about 2 days to about 10 days following use of desmopressin, from about 1 days to about 12 days following use of desmopressin, from about 30 minutes to about 60 minutes following use of desmopressin.

[0142] 120. The use of any one of embodiments 109 to 119, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

[0143] 121. The use of any one of embodiments 109 - 120, wherein the subject is a human.

[0144] 122. Use of desmopressin prior to surgery to remove some or all of a glioblastoma in a subject, preferably about 30 minutes to about 60 before surgery for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma;

[0145] - use of desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma;

[0146] - use of temozolomide at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy;

[0147] - use of temozolomide for 6 cycles of maintenance treatment; and

[0148] - use of desmopressin on day 1 of each said cycle.

[0149] 123. The use of embodiment 122, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

[0150] 124. The use of embodiment 122 or 123, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

[0151] 125. The use of any one of embodiments 122 to 124, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

[0152] 126. The use of any one of embodiments 122 to 125, wherein temozolomide is for administration at a dose of from 75 to 200 mg / m2daily.

[0153] 127. The use of any one of embodiments 122 to 126, wherein said subject is a human.BRIEF DESCRIPTION OF THE FIGURES

[0154] Embodiments of the present disclosure will now be described, byway of example only, with reference to the attached Figures.

[0155] Figure 1. Evaluation of desmopressin (dDAVP) on LN229 Clonogenic Growth. The effect of desmopressin (dDAVP) on the clonogenic growth of LN229 glioma cells was evaluated using a 2D colony formation assay. LN229 cells were treated with a range of dDAVP concentrations (0.1-1 pM) or vehicle control for 7 d in complete medium, with medium changes occurring every 72 h. After treatment, colonies were fixed, stained, and quantified to determine the impact of dDAVP on the ability of cancer cells to grow at low density. Two independent experiments. n=6-12 per experiment. ANOVA plus Tukey's multiple comparisons test. ***p<0.001; ****p<0.0001

[0156] Figure 2. Confirmation of dDAVP Effects on A172 Clonogenic Growth. The effect of desmopressin (dDAVP) on the clonogenic growth of glioma cells was confirmed using A172 cells. A 2D colony formation assay was carried out using A172 cells treated with 1 pM dDAVP or vehicle control for 7 d in complete medium, with medium changes occurring every 72 h. Two independent experiments. n=6-12 per experiment. Unpaired ttest; ****p<0.0001.

[0157] Figure 3. Effect of dDAVP on A172 Cell Migration. The impact of desmopressin (dDAVP) on the migration of A172 glioma cells was evaluated using a transwell migration assay. A172 cells were treated with 1 pM dDAVP or vehicle control, and their migration was assessed over a 17h incubation period. Migrating cells were fixed, stained and washed. 4-5 randomly selected X400-high power fields (HPF) were quantified por insert using QuPath v0.3.0 and normalized to control. Three independentexperiments, n=8-10 X400-HPF per experimental group per replicate. Unpaired t test. ***p<0.001 .

[0158] Figure 4. Effect of dDAVP on LN229 Cell Migration. The impact of desmopressin (dDAVP) on the migration of LN229 glioma cells was evaluated using a transwell migration assay. Cells were treated with 1 pM dDAVP or vehicle control, and their migration was assessed over a 17h incubation period. Migrating cells were fixed, stained and washed. 4-5 randomly selected X400-high power fields (HPF) per insert were quantified using QuPath v0.3.0 and normalized to control. Three independent experiments, n=8-10 X400-HPF per experimental group per replicate. Unpaired t test. P>0.05 (non-significant)

[0159] Figure 5. Log-Phase Proliferation of LN229 Cells treated with dDAVP Treatment. The effects of dDAVP on the log-phase proliferation of LN229 glioma cells were assessed using a standard high-density cell proliferation assay. LN229 cells were treated with a range of dDAVP concentrations (0.1-10 pM) or vehicle control, and proliferation was assessed using the crystal violet staining method. Two independent experiments, n=6-12 per experimental group per independent replicate. ANOVA cont. Tukey's multiple comparisons test; *p<0.05, **p<0.01 and ***p<0.001.

[0160] Figure 6. Log-Phase Proliferation of 172 Cells with dDAVP Treatment. The effects of dDAVP on the log-phase proliferation of A172 glioma cells were assessed using a standard high-density cell proliferation assay. A172 cells were treated with a range of dDAVP concentrations (0.1-10 pM) or vehicle control, and proliferation was assessed using the MTS method. Two independent experiments, n=6-12 per experimental group per independent replicate. ANOVA cont. Tukey's multiple comparisons test; **p<00.1 and ****p<0.0001.

[0161] Figure 7. AVPR2 Gene Expression in GBM Cells. The expression of the AVPR2 gene in GBM cells was assessed using quantitative polymerase chain reaction (qPCR). Total RNA was purified from LN229 and A172 GBM cell lines, as well as from MG-63 (human AVPR2-expressing osteosarcoma, positive control) and U-2 OS (human AVPR2-negative osteosarcoma, negative control) cells. Following cDNA synthesis, qPCR was performed, with Ct values normalized to HPRT1 expression levels. Each sample was analyzed in triplicate, and AVPR2 expression in MG-63 cells was set as the reference at “100%.” Relative quantification values were calculated using the formula 2A(-AACt). ANOVA cont. Tukey's multiple comparisons test. ***p<0.001.

[0162] Figure 8. Immunohistochemical Staining for AVPR2 in GBM Xenografts. Immunohistochemical staining for AVPR2 in LN229 xenografts was conducted. Tissue sections from GBM xenografts were analyzed to assess the expression and localization of AVPR2. (A) anti-AVPR2 staining shows strong focal immunoreactivity, (B) anti-AVPR2 showing moderate immunoreactivity, (C) anti-AVPR2 showing weak and “patchy” immunoreactivity, (D) anti-AVPR2 showing weak / moderate focal immunoreactivity in GBM, (E) Positive Control (MG-63 hOSA xenograft, moderate immunoreactivity), (F) Negative Control (Omission of 1ary mAb anti-AVPR2).

[0163] Figure 9. Bioinformatic Assessment of AVPR2 Impact on Overall Survival in GBM. A bioinformatic analysis of AVPR2 expression and its association with overall survival (OS) in GBM patients was conducted, utilizing data from the TIMER2.0 database. The graph displays the survival curves for patients grouped by AVPR2 expression levels across all GBM subtypes: low AVPR2 expression levels (blue line) vs. high AVPR2 expression levels (red line). Log rank test, p<0.001. HR<1.

[0164] Figure 10. Bioinformatic Assessment of AVPR2 expression in GBM (A) and LGG (B) versus normal tissue. A bioinformatic analysis of AVPR2 expression in GBM and LGG (grey) versus normal tissue (red) was conducted, using (TCGA / GTEx). GEPIA2 Database.

[0165] Figure 11. Correlation of AVPR2 Expression with Immune Infiltrates in GBM tumours. The correlation between AVPR2 expression and immune cell infiltrates in GBM, specifically CD8+ T cells, natural killer (NK) cells, and B cells, in the TME was analyzed using deconvolution methods applied to RNA sequencing data from the TIMER2.0 platform (TCGAGBM database).

[0166] Figure 12. Correlation of AVPR2 Expression with Immune Infiltrates in LGG tumours. The correlation between AVPR2 expression and immune cell infiltrates in LGG, specifically CD8+ and CD4+ T cells, natural killer (NK) cells, and B cells, in the TME was analyzed using deconvolution methods applied to RNA sequencing data from the TIMER2.0 platform (TCGAGBM database)

[0167] Figure 13. Effect of Temozolomide (TMZ) on Long-Term Clonogenic Growth of GBM Cultures as monotherapy. The effect of TMZ on the long-term growth of GBM cultures was evaluated. GBM cells were treated with varying concentrations of (10-100 pM) over 7 d. Two independent experiments. n=6 per experimental group per independent replicate. ANOVA plus Dunnett's multiple comparisons test. ****p<0.0001

[0168] Figure 14. Combinational Treatment Schemes for TMZ and dDAVP. Two distinct in vitro treatment schemes were conducted to evaluate the effects of combining dDAVP and TMZ on GBM cells. The first scheme involves sequential exposure, where cells are treated with dDAVP alone from day 0 to day 3, followed by TMZ alone from day 3 to day 7. The second scheme features concomitant exposure, with cells receiving both dDAVP and TMZ simultaneously from day 3 to day 7.

[0169] Figure 15. Combination of dDAVP and TMZ in Long-Term Clonogenic Growth of GBM Cultures. The combination of dDAVP and TMZ on the long-term clonogenic growth of GBM cultures, following the treatment schemes outlined in Figure 14 was conducted. GBM cells were exposed to vehicle control, 1 pM dDAVP, 10 pM TMZ, or the combination of both drugs, utilizing either a sequential (15A) or concomitant (15B) treatment approach. Two independent experiments. n=6 per experimental group per independent replicate. ANOVA plus Dunnett's multiple comparisons test, *p<0.05, ***p<0.001 and ****p<0.0001.

[0170] Figure 16. Depicts one example of the use of desmopressin in GBM care continuum.

[0171] Figure 17. Preclinical protocol design: Administration of dDAVP in the peritumour implantation window, alone or followed by adjuvant TMZ.

[0172] Figure 18. Preclinical protocol design: Co-adjuvant use of dDAVP with concomitant TMZ-based chemotherapy.

[0173] Figure 19. Effect of early administration of dDAVP, alone or followed by adjuvant TMZ, on GBM xenograft progression. (A) GBM tumour volume overtime. Values expressed as mean±SEM. (B) GBM tumour volume at day 36. Data expressed as scattered dot blot and mean. (C) GBM tumour volume at day 43. Data expressed as scattered dot blot and mean. (D) GBM tumour volume at day 51. Data expressed as scattered dot blot and mean. (E) GBM tumour volume at day 58. Data expressed as scattered dot blot and mean. n=5 animals per experimental group. ANOVA plus Tukey's multiple comparisons test, *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001.

[0174] Figure 20. Impact of dDAVP, alone or in addition to TMZ-based chemotherapy, on GBM tumour growth dynamics. (A) Representation of individualized tumour growth rates obtained between days 26 and 58 of the GBM progression protocol. (B) Mean GBM tumour growth rates obtained between days 26 and 58 of the xenograft progression protocol. Values expressed as mean±SD. For a better visualization of results, the left "y" axis is presented as two segments with different scales (Bottom segment from-3 to 2, with major ticks every 3 points; Top segment from 10 to 50, with majors ticks every 10 points). n=5 animals per experimental group. ANOVA plus Tukey's multiple comparisons test, ****p<0.0001.

[0175] Figure 21. Representative photographs of GBM xenograft-bearing mice from different experimental groups at day 56

[0176] Figure 22. Body weight of the animals throughout the protocol. (A) Representation of body weight of the animals belonging to the different experimental groups. Values expressed as mean±SD. (B) Representation of body weight of the animals belonging to the different experimental groups, relative to the values of day 0. Values expressed as mean±SD. n=5 animals per experimental group. ANOVA plus Tukey's multiple comparisons test, non-significant p>0.05.

[0177] Figure 23. Therapeutic effect of dDAVP addition to adjuvant TMZ on GBM xenograft progression. (A) GBM tumour volume overtime. Values expressed as mean±SEM. (B) GBM tumour volume at day 26. Data expressed as scattered dot blot and mean. (C) GBM tumour volume at day 36. Data expressed as scattered dot blot and mean. (D) GBM tumour volume at day 40. Data expressed as scattered dot blot and mean. (E) GBM tumour volume at day 51. Data expressed as scattered dot blot and mean. n=5 animals per experimental group. ANOVA plus Tukey's multiple comparisons test, *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001.

[0178] Figure 24. Impact of dDAVP addition to adjuvant TMZ on GBM tumour growth dynamics. (A) Representation of individualized tumour growth rates obtained between days 19 and 51 of the GBM progression protocol. (B) Mean GBM tumour growth rates obtained between days 19 and 51 of the xenograft progression protocol. Values expressed as mean±SD. For a better visualization of results, the left "y" axis is presented as two segments with different scales (Bottom segment from -2 to 1.5, with major ticks every 2 points; Top segment from 10 to 30, with majors ticks every 5 points). n=5 animals per experimental group. ANOVA plus Dunnett's multiple comparisons test, *p<0.05, ***p<0.001 and ****p<0.0001.

[0179] Figure 25. Representative photographs of GBM xenograft-bearing mice from different experimental groups at day 50.

[0180] Figure 26. Body weight of the animals throughout the protocol. (A) Representation of body weight of the animals belonging to the different experimental groups. Values expressed as mean±SD. (B) Representation of body weight of the animals belonging to the different experimental groups, relative to the values of day 0.Values expressed as mean±SD. n=5 animals per experimental group. ANOVA plus Tukey's multiple comparisons test, non-significant p>0.05.DETAILED DESCRIPTION

[0181] In one aspect, there is provided a method of treating a cancer, glioblastoma, in a subject in need thereof.

[0182] In one aspect, the subject has glioblastoma, is suspected of having glioblastoma, or is at risk of developing glioblastoma.

[0183] Cancer

[0184] The term “cancer”, as used herein, refers to a variety of conditions caused by the abnormal, uncontrolled growth of cells. Cells capable of causing cancer, referred to as “cancer cells”, possess characteristic properties such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and / or certain typical morphological features. Cancer cells may be in the form of a tumour, but such cells may also exist alone within a subject, or may be a non-tumourigenic cancer cell. A cancer can be detected in any of a number of ways, including, but not limited to, detecting the presence of a tumour or tumours (e.g., by clinical or radiological means), examining cells within a tumour or from another biological sample (e.g., from a tissue biopsy), measuring blood markers indicative of cancer, and detecting a genotype indicative of a cancer. However, a negative result in one or more of the above detection methods does not necessarily indicate the absence of cancer, e.g., a patient who has exhibited a complete response to a cancer treatment may still have cancer, as evidenced by a subsequent relapse. In a specific example, the cancer is glioblastoma.

[0185] Glioblastoma

[0186] Glioblastoma (GMB) is the most common and aggressive primary CNS tumour in adults.

[0187] Glioblastoma was historically defined as a Grade IV astrocytoma under the WHO Classification of CNS Tumors (2000 and 2007 editions). Key diagnostic criteria included: high mitotic activity, pseudopalisading necrosis, microvascular proliferation, cellular pleomorphism with nuclear atypia.

[0188] Glioblastomas have been classified as: (i) Primary glioblastoma (de novo): Occurring without precursor lesions, more common in older adults, and (ii) Secondary glioblastoma: Progressing from lower-grade gliomas, typically affecting younger patients.

[0189] In some examples, “glioblastoma” refers to an aggressive brain tumour belonging to Grade IV astrocytoma brain tumour. The term glioblastoma also includes its variants gliosarcoma, giant cell glioblastoma and small cell glioblastoma. Because cells in this tumour vary in size and shape, i.e. they are pleomorphic, glioblastoma is also called glioblastoma multiforme. In some examples, the glioblastoma is a primary cancer. In another example, the glioblastoma returns after a period of remission, which may be referred to as recurrent glioblastoma.

[0190] The 2021 WHO classification of glioblastoma generally refines its diagnostic criteria, as compared to earlier classifications, by excluding tumours with better prognoses (e.g., IDH-mutant astrocytomas) and emphasizing the aggressive molecular profile of IDH-wildtype glioblastomas.

[0191] This reclassification focuses on glioblastoma’s high mortality rate and need for targeted therapies.

[0192] The WHO Classification of CNS Tumors (2021) emphasize a combined histological and molecular classification. Glioblastoma may also be classified as an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, +7 / -10 chromosomal signature.

[0193] In some examples, glioblastomas may be identified using biomarkers and genetic mutations.

[0194] In the 2021 WHO guidelines, glioblastoma is identified as an IDH-wildtype diffuse astrocytic tumour of WHO Grade IV.

[0195] The inclusion criteria for glioblastoma may include one or more of the following: TERT promoter mutations, EGFR amplification, +7 / -10 chromosomal signature.

[0196] In some examples IDH-mutant glioblastomas may be classified as Grade IV astrocytomas.

[0197] IDH-wildtype is predominantly associated with aggressive glioblastomas in older adults.

[0198] IDH-mutant is associated with better prognosis.

[0199] In some examples, MGMT promoter methylation may be used as a predictor of responsiveness to temozolomide therapy. EGFR amplification and TERT promoter mutations are linked to the tumour’s aggressive behavior.

[0200] Treatment of glioblastoma

[0201] In one aspect, there is provided a method of treating a glioblastoma in a subject in need thereof comprising, or consisting of, administering a therapeutic amount of desmopressin. In one aspect, the subject has glioblastoma, is suspected of having glioblastoma, or is at risk of developing glioblastoma.

[0202] Desmopressin and vasopressin analogues

[0203] As used herein, the term “desmopressin”, “DDAVP®”, or “dDAVP”, a vasopressin analogue, refers to 1-desamino-8-D-arginine vasopressin.

[0204] Desmopressin is a synthetic derivative of vasopressin (AVP) that primarily acts as a selective agonist for the arginine vasopressin type 2 receptor (AVPR2), which is expressed in microvascular endothelial cells. Originally developed for its hemostatic effects, desmopressin induces the release of von Willebrand factor and Factor VIII, enhancing blood coagulation and promoting vascular normalization.

[0205] In one embodiment, the vasopressin analogue, such as desmopressin, is formulated as a pharmaceutically acceptable salt, such as desmopressin acetate. In a further embodiment, the vasopressin analogue is formulated in a pharmaceutical composition, comprising the vasopressin analogue and a pharmaceutically acceptable excipient and / or carrier. In one embodiment, the vasopressin analogue is desmopressin.

[0206] In one aspect, there is provided a method of treating a glioblastoma in a subject in need thereof comprising, or consisting of, administering a therapeutic amount of a vasopressin analogue. In one aspect, the subject has glioblastoma, is suspected of having glioblastoma, or is at risk of developing glioblastoma, comprising, administering a therapeutic amount of a vasopressin analogue.

[0207] The term “vasopressin analogue” as used herein, refers to compounds or derivatives having a similar function to vasopressin but not necessarily a similar structure, and includes all compounds or derivatives having anti-proliferative activity, including prodrugs.

[0208] Vasopressin analogues include, but are not limited to, desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0209] In one aspect, there is provided a method of treating a glioblastoma in a subject in need thereof comprising, or consisting of, administering a therapeutic amount of a vasopressin analogue. In one aspect, the subject has glioblastoma, is suspected of having glioblastoma, or is at risk of developing glioblastoma, comprising, administering a therapeutic amount of a vasopressin analogue. In a specific example, the vasopressinanalogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0210] AVPR2

[0211] In one aspect, there is provided a method for treating or preventing glioblastoma in a subject, comprising, or consisting of, measuring an amount of AVPR2 polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from the subject; and administering to the subject in need thereof a therapeutically effective amount of desmopressin, when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

[0212] The term "sample" as used herein refers to any sample from a subject, including but not limited to a fluid, cell or tissue sample that comprises cancer cells, or which is suspected of containing cancer cells, which can be assayed for gene expression levels, proteins levels, enzymatic activity levels, and the like. The sample may include, for example, a brain sample, a blood sample, a fractionated blood sample, a bone marrow sample, a biopsy, a frozen tissue sample, a fresh tissue specimen, a cell sample, and / or a paraffin embedded section, material from which RNA can be extracted in sufficient quantities and with adequate quality to permit measurement of relative mRNA levels, or material from which polypeptides can be extracted in sufficient quantities and with adequate quality to permit measurement of relative polypeptide levels.

[0213] In one example, a sample containing cancerous cells or suspected of containing cancerous cells is obtained from a subject with cancer. Collection of such a sample is well known to the skilled worker. In a specific example, the sample is a blood sample. Methods of obtaining a sample, processing and / or storage of such a sample are also well known to the skilled worker.

[0214] In another example, a method of the present disclosure comprises qualitatively or quantitatively determining, analyzing or measuring arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in sample from a subject suspect of having glioblastoma, at risk of developing glioblastoma, or having glioblastoma.

[0215] In one example, in which a patient sample is determined to have a high amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, optionally compared to a control, the patient is identified as a candidate for treatment with desmopressin, as described herein.

[0216] Surgical and Radiotherapeutic Treatment of Glioblastoma

[0217] Currently, the conventional treatment for GBM is surgery to remove some or all of a glioblastoma is a patient, followed by radiation and chemotherapy.

[0218] A primary objective of surgery is to remove as much of the GBM tumour as possible while minimizing injury to the surrounding normal brain tissue which is needed for normal neurological function. However, GBMs are surrounded by a zone of migrating, infiltrating tumour cells that invade surrounding tissues, making it difficult or impossible to remove the tumour entirely. Surgery provides the ability to reduce the amount of solid tumour tissue within the brain, remove those cells in the center of the tumour that may be resistant to radiation and / or chemotherapy and reduce intracranial pressure. Surgery, by providing a debulking of the tumour, carries the ability to prolong the lives of some patients and improve the quality of remaining life.

[0219] Typically, the surgery is a craniotomy, frequently with computer-assisted image-guidance and at times using intra-operative mapping techniques to determine the locations of the motor, sensory and speech / language cortex. Intraoperative mapping may involve operating on a patient while they are awake and mapping the anatomy of their language function during the operation.

[0220] The term "resection" refers to the surgical removal or excision of part or all of a tumour, such as a glioblastoma.

[0221] Radiation therapy follows surgery. Currently, in standard external beam radiation therapy, multiple sessions of standard-dose “fractions” of radiation are delivered to the tumour site as well as a margin in order to treat the zone of infiltrating tumour cells. Each treatment induces damage to both healthy and cancerous tissue.

[0222] Radiosurgery is a treatment using radiation delivery systems to focus radiation at the site of the tumour, while minimizing the radiation dose to the surrounding brain. Radiosurgery may be used in select cases for tumour recurrence, often using additional information derived from MRS or PET scans. It is rarely used in the initial treatment of GBM.

[0223] Chemotherapy with temozolomide is the current standard of treatment for GBM. In one example, temozolomide is generally administered every day during radiation therapy and then for six cycles after radiation during the maintenance phase.

[0224] Radiotherapy

[0225] In some examples, in radiotherapy of high-grade gliomas (HGG) are based on anatomic imaging techniques, usually computed tomography (CT) scanning andmagnetic resonance imaging (MRI). The guidelines vary depending the type and / or stage of the glioblastoma.

[0226] Desmopressin and temozolomide

[0227] In another aspect, the method of treating glioblastoma further comprises, or consists of, administering desmopressin and temozolomide to a subject having glioblastoma, at risk of developing glioblastoma, or believed to have glioblastoma.

[0228] Temozolomide

[0229] As used herein, “temozolomide”, also referred to as TMZ and TEMODAL®, refers to a triazines alkylating agent and more specifically compound of formula 3,4-Dihydro-3-methyl-4-oxoimidazo[5,1-d][1 ,2,3,5]tetrazine-8-carboxamide and pharmaceutically acceptable salts thereof (CAS number of 85622-93-1). Alkylating agents directly damage DNA (the genetic material in each cell) to keep the cell from reproducing.

[0230] As used herein, the term "co-administer" is intended to embrace separate administration of a vasopressin analogue, such as desmopressin, and temozolomide in a sequential manner as well as co-administration of these agents in a substantially simultaneous manner, such as in a single mixture / composition or in doses given separately, but nonetheless administered substantially simultaneously to the subject. In one example, a subject is first administered desmopressin, followed by administration with temozolomide. In one example, a subject is first administered with temozolomide followed by administration with desmopressin.

[0231] In one aspect, there is provided a method of treating a glioblastoma in a subject in need thereof comprising, or consisting of, administering a therapeutic amount of desmopressin and temozolomide. In one aspect, the subject has glioblastoma, is suspected of having glioblastoma, or is at risk of developing glioblastoma.

[0232] A subject at risk of developing glioblastoma may be an individual at risk of developing glioblastoma, such as a subject in remission, having a family history, and / or having some other predisposition.

[0233] In an embodiment of the disclosure, the vasopressin analogue is administered at a dose effective to reduce or halt proliferation of the cancer cells, for example by inducing cell cycle arrest. In one embodiment, the cancer cells are glioblastoma cells and the vasopressin analogue is desmopressin.

[0234] In another embodiment of the disclosure, the vasopressin analogue is administered at a dose effective to reduce or halt cancerous tumour growth, and / ordecrease tumour volume. In one embodiment, the cancerous tumour growth is a glioblastoma and the vasopressin analogue is desmopressin.

[0235] In another embodiment of the disclosure, the vasopressin analogue is administered at a dose effective to reduce or prevent metastases of cancer cells to other tissues and organs of the subject’s body. In one embodiment, the cancer cells are glioblastoma cells and the vasopressin analogue is desmopressin.

[0236] In another embodiment, desmopressin reduces tumour cell aggregation and early engraftment.

[0237] In another embodiment, desmopressin reduces the rate of tumour growth.

[0238] In another embodiment, desmopressin reduces tumour cell plasticity and / or invasive phenotype.

[0239] In another embodiment of the disclosure, the vasopressin analogue is coadministered with temozolomide, at a dose which enhances the anti-proliferative efficacy of the vasopressin analogue. In one embodiment, the vasopressin analogue is desmopressin.

[0240] In one aspect, there is provided a method of treating a glioblastoma in a subject in need thereof comprising, or consisting of, administering a therapeutic amount of a vasopressin analogue and temozolomide. In one aspect, the subject has glioblastoma, is suspected of having glioblastoma, or is at risk of developing glioblastoma, comprising, administering a therapeutic amount of a vasopressin analogue and temozolomide. In a specific example, the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

[0241] In one aspect, there is provided a method for treating or preventing glioblastoma in a subject, comprising, or consisting of, measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject; and administering to the subject in need thereof a therapeutically effective amount of desmopressin, optionally in a physiological carrier or a pharmaceutically acceptable salt thereof, when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

[0242] In one aspect, the method further comprising, or consisting of, administering a therapeutically effective amount of Temozolomide (TMZ).

[0243] In one embodiment, the vasopressin analogue, for example desmopressin acetate, is formulated as an aqueous solution of desmopressin acetate present at a concentration of about 4 pg / ml or about 15 pg / ml.

[0244] In one embodiment, desmopressin is administered at a dose of between about 0.2 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 pg / kg to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg. In one example, desmopressin is administered daily. In one embodiment, desmopressin is formulated for intravenous, intramuscular or subcutaneous administration. In one embodiment, desmopressin is formulated for immediate release, IV infusion, delayed release or depot administration.

[0245] In one embodiment, a vasopressin analogue administered at a dose of of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg. In one example, the vasopressin analogue is administered daily. In one embodiment, the vasopressin analogue is formulated for intravenous, intramuscular or subcutaneous administration. In one embodiment, the vasopressin analogue is formulated for immediate release, IV infusion, delayed release or depot administration.

[0246] In one example, the subject is first treated with desmopressin, followed by treatment with TMZ. In one example, desmopressin is first administered to the subject followed by TMZ treatment. In one example, TMZ is administered from about 3 days to about 7 days following desmopressin administration. In another example, TMZ is administered from about 2 days to about 10 days following desmopressin administration. TMZ is administered from about 1' days to about 12 days following desmopressin administration. TMZ is administered from about 30 minutes desmopressin administration.

[0247] In one example, desmopressin may administered before, during and / or after tumour surgery of the glioblastoma.

[0248] In one example, desmopressin may be administered perioperatively.

[0249] In one embodiment, TMZ is administered at a dose of from 75 to 200 mg / m2daily.

[0250] In one example, desmopressin is first administered to the subject followed by TMZ treatment. In one example, TMZ is administered 2 to 4 weeks following desmopressin administration.

[0251] Desmopressin in combination with surgery and temozoloimde

[0252] In one aspect example, desmopressin may be used in combination with other treatments of glioblastoma, including surgery and temozolomide, in the treatment of glioblastoma.

[0253] Figure 16 depicts one example of the use of desmopressin in the GBM care continuum.

[0254] In one example, a treatment of glioblastoma comprises an (i) Initiation Phase, and a (ii) Maintenance Phase.

[0255] Surgical removal of glioblastoma

[0256] In one example, in a subject having glioblastoma, desmopressin may administered prior to surgery, preferably about 30 minutes before surgery, said surgery to remove some or all of the glioblastoma. Desmopressin may then administered following surgery, preferably about 24 hour following surgery.

[0257] Initiation Phase / concomitant chemotherapy and radiotherapy

[0258] Following surgery, the patient undergoes the standard of treatment for glioblastoma. In one example the standard of treatment is administration of temozolomide at a dose of 75 mg / m2daily for 42 days concomitant with radiotherapy (preferably 60 Gy administered in 30 fractions).

[0259] About four weeks following standard treatment, maintenance phase I maintenance therapy may be started.

[0260] Maintenance phase / maintenance therapy

[0261] Four weeks after completing the standard treatment, temozolomide is administered for an additional 6 cycles of maintenance treatment. Dosage in Cycle 1 (maintenance) is 150 mg / m2once daily for 5 days followed by 23 days without treatment. At the start of Cycle 2, the dose is escalated to 200 mg / m2, if the CTC non-hematologic toxicity for Cycle 1 is Grade <2 (except for alopecia, nausea and vomiting), absolute neutrophil count (ANC) is >1.5 x 109 / L, and the platelet count is >100 x 109 / L. If the dose was not escalated at Cycle 2, escalation should not be done in subsequent cycles. The dose remains at 200 mg / m2per day for the first 5 days of each subsequent cycle except if toxicity occurs.

[0262] Desmopressin is administered on day 1 of each cycle.

[0263] Recurrence / progression

[0264] In the case or recurrence or progression of glioblastoma, in patients previously untreated with chemotherapy, temozolomide is administered at a dose of 200 mg / m2once daily for 5 days per 28-day cycle. For patients previously treated with chemotherapy, the initial dose is 150 mg / m2once daily for 5 days, to be increased in the second cycle to 200 mg / m2once daily for 5 days, providing there is no hematologic toxicity.

[0265] Desmopressin may be administered every 28 days, prior to the start of each temozolomide cycle.

[0266] The term “therapeutically effective amount”, as used herein, refers to an amount that is effective for preventing, ameliorating, or treating a disease or disorder (e.g., glioblastoma).

[0267] The term “pharmaceutically effective amount” as used herein refers to the amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by a researcher or clinician. This amount can be a therapeutically effective amount.

[0268] The term “pharmaceutically acceptable” as used herein refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0269] The term “pharmaceutically acceptable carrier” as used herein refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting the subject agents from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, for example the carrier does not decrease the impact of the agent on the treatment. In other words, a carrier is pharmaceutically inert. The terms “physiologically tolerable carriers” and “biocompatible delivery vehicles” are used interchangeably. Thus, the term “carrier” or “excipient” may refer to a non-toxic solid, semi-solid or liquid filler, diluent. The term includes solvents, dispersion, media, coatings, isotonic agents, and adsorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art.

[0270] Some examples of materials that may serve as pharmaceutically acceptable carriers include, but are not limited to, sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butterand suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol;phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the subject. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound useful within the invention.

[0271] The term “pharmaceutically acceptable salt" refers, for example, to a salt that retains the desired biological activity of a compound of the present disclosure and does not impart undesired toxicological effects thereto; and may refer to an acid addition salt or a base addition salt.

[0272] As used herein, the term “pharmaceutically-acceptable salts” refers to the conventional nontoxic salts or quaternary ammonium salt. These salts can be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting a compound in its free base or acid form with a suitable organic or inorganic acid or base, and isolating the salt thus formed during subsequent purification. Conventional nontoxic salts include those derived from inorganic acids such as sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.

[0273] As used herein, the term “prodrug” refers to a substance that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of, for example, endogenous enzymes or other chemicals and / or conditions. Prodrug derivatives of desmopressin, or pharmaceutically acceptable salts or solvates thereof, can be prepared by methods known to those of ordinary skill in the art.

[0274] As used herein, a “subject” refers to all members of the animal kingdom including mammals, and suitably refers to humans. A member of the animal kingdom includes, without limitation, a mammal (such as a human, primate, swine, sheep, cow, equine, horse, camel, canine, dog, feline, cat, tiger, leopard, house pet, livestock, rabbit, mouse, rat, guinea pig or other rodent, seal, whale and the like). In an embodiment of thepresent disclosure, the subject is in need of a treatment of the disclosure. In one example, the subject is a human.

[0275] The term “treatment”, “treat”, or “treating” as used herein, refers to obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. "Treating" and "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0276] The term "amelioration" or "ameliorates" as used herein refers to a decrease, reduction or elimination of a condition, disease, disorder, or phenotype, including an abnormality or symptom.

[0277] The term "symptom" of a disease or disorder is any morbid phenomenon or departure from the normal in structure, function, or sensation, experienced by a subject and indicative of disease.

[0278] As used herein, the term “neoadjuvant” refers to an initial treatment of a disease in a subject, and thus a “neoadjuvant” cancer refers to a cancer in a patient that has not received prior treatment to a therapeutic compound of composition as described herein.

[0279] A "treatment regimen" as used herein refers to a combination of dosage, frequency of administration, or duration of treatment, with or without addition of a second medication.

[0280] For example, a subject with glioblastoma may be treated to prevent progression or alternatively a subject in remission can be treated with a compound or composition described herein to prevent recurrence.

[0281] As used herein, the term “metastatic” is defined as the transfer of cancer cells from one organ or part to another not directly connected with it.

[0282] The term "therapeutically effective amount" as used herein means an amount effective, at dosages and for periods of time necessary to achieve the desired result. Effective amounts may vary according to factors such as the disease state, age, sex and / or weight of the subject. The amount of a given compound or composition that will correspond to such an amount will vary depending upon various factors, such as thegiven drug or compound, the pharmaceutical formulation, the route of administration, the identity of the subject being treated, and the like, but can nevertheless be routinely determined by one skilled in the art.

[0283] A “pharmaceutical composition” as used herein refers to a chemical or biological composition suitable for administration to a subject. Such compositions may be specifically formulated for administration via one or more of a number of routes.

[0284] The term “administered” or “administering” as used herein means administration of a therapeutically effective dose of a compound or composition of the disclosure to a subject.

[0285] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing the active compound into association with a carrier, which may constitute one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active compound with liquid carriers or finely divided solid carriers or both, and then if necessary, shaping the product.

[0286] The compounds and compositions may be administered to a subject by any convenient route of administration, whether systemically / peripherally or at the site of desired action, including but not limited to, oral (e.g. by ingestion); topical (including e.g. transdermal, intranasal, ocular, buccal, and sublingual); pulmonary (e.g. by inhalation or insufflation therapy using, e.g. an aerosol, e.g. through mouth or nose); rectal; vaginal; parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intra-arterial, intra-cardiac, intrathecal, intra-spinal, intra-capsular, sub-capsular, intra-orbital, intraperitoneal, intra-tracheal, subcuticular, intraarticular, subarachnoid, and intra-sternal; by implant of a depot I for example, subcutaneously or intramuscularly.

[0287] In understanding the scope of the present disclosure, the term "comprising" and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, "including", "having" and their derivatives. Finally, terms of degree such as "substantially", "about" and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result isnot significantly changed. These terms of degree should be construed as including a deviation of at least 5% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0288] The terms “peptide”, “polypeptide,” and “protein”, as used herein are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein's or peptide's sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, ora combination thereof.

[0289] The term “antibody” or “antibodies” is used herein refers to both polyclonal and monoclonal antibodies. In addition to intact or “full” immunoglobulin molecules, also included in the term “antibodies” are fragments (e.g., CDRs, Fv, Fab and Fc fragments) or polymers of those immunoglobulin molecules and humanized versions of immunoglobulin molecules, as long as they exhibit any of the desired properties according to the description.

[0290] In another embodiment of the disclosure, there is included a kit for the treatment of glioblastoma.

[0291] In one embodiment, the disclosure includes a kit for the treatment of glioblastoma, comprising, or consisting of, (i) a therapeutically effective amount of desmopressin, and (ii) a therapeutically effect amount of a TMZ; and instructions for using the kit. In one embodiment, the vasopressin analogue is desmopressin.

[0292] In one embodiment, the disclosure includes a kit for the treatment of glioblastoma, comprising, or consisting of, (i) a therapeutically effective amount of a vasopressin analogue, and (ii) a therapeutically effect amount of a TMZ; and instructions for using the kit. In one embodiment, the vasopressin analogue is desmopressin,synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin..

[0293] To gain a better understanding of the invention described herein, the following examples are set forth. It should be understood that these examples are for illustrative purposes only. Therefore, they should not limit the scope of this invention in anyway.

[0294] EXAMPLES

[0295] EXAMPLE 1

[0296] Materials and methods

[0297] Drugs and reagents

[0298] Injectable (20 pg / ml) dDAVP acetate (MW 1129.3 g / mol), medical degree, was obtained from Biofactor SA (Buenos Aires, Argentina). TMZ (MW 194, 151 g / mol) was obtained from Microsules Argentina SA (Buenos Aires, Argentina). The compound was initially dissolved in dimethyl sulfoxide (DMSO) to generate concentrated stocks. Subsequent dilutions using PBS were carried out to attain final working concentrations. PBS or appropriate concentrations of DMSO, respectively, were used as vehicles for the control groups.

[0299] Cell lines and culture conditions

[0300] LN229 (ATCC CRL-2611 ™) is a highly aggressive human GBM cell model, widely used in biomedical research. It was isolated in 1979 from the right frontal parieto-occipital cortex of a White, 60-year-old, female patient with GBM. It has an epithelial-like morphology and it is tumourigenic in nude mice. It bears a mutation in p53 (TP53) and possible homozygous deletions in the p16 and p14ARF tumour suppressor genes. LN229 mimics primary, IDH-wildtype glioblastomas, particularly those with resistance to standard therapies.

[0301] Human GBM A172 (ATCC CRL-1620 ™) cell line was used as an additional experimental model for selected assays. It was isolated from the brain tissue of a 53-year-old, male patient with GBM. It has a fusiform morphology, it is highly migratory and invasive, and expresses genes related to mesenchymal markers. It is not tumourigenic in immunosuppressed mice. A172 models IDH-wildtype glioblastomas with receptor tyrosine kinase pathway activation and mesenchymal features, consistent with invasive glioblastoma subtypes.

[0302] LN229 and A172 were grown using Dulbecco's modified Eagle's medium (DMEM) culture medium (Gibco, USA) and Roswell Park Memorial Institute medium (RPMI), respectively, with 10% FBS (Natocor, Argentina), 80 pg / ml gentamicin, and 2mM glutamine, in monolayer culture, at 37°C in a humidified atmosphere of 5% CO2. Cells were harvested using a trypsin / EDTA solution (Thermo Fisher Scientific Inc., USA) diluted in PBS and routinely tested for mycoplasma.

[0303] GBM clonogenic growth

[0304] Clonogenic assays were conducted in order to evaluate the effect of dDAVP on LN229 and A172 cell 2D GBM colony formation. GBM cell seeding was conducted at low density (300-800 cells / well) using 96-well flat bottom plates. Cells were cultured for 7 days in complete medium which was replaced every 72 h with dDAVP (0.1-1 pM) or vehicle. At the end of the week, GBM colonies were methanol-fixed and stained with 0.5% crystal violet solution to facilitate quantification. Representative pictures of each well were obtained from the scanned plate, by the Epson Image Scanner III (GE Healthcare, Bio-Sciences AB, Sweden) and colony quantification was conducted per well using high-resolution scanning and computer assisted image analysis using the open source QuPath Software v0.3.0, and normalized to control (QuPath, Quantitative Pathology and Bioimage Analysis, University of Edinburgh, UK, qupath.github.io). Performed by duplicate (independent operators), 6-12 wells per experimental group per independent replicate.

[0305] GBM cell migration

[0306] Transwell® migration assays were conducted to evaluate GBM cell chemotaxis. After overnight starvation (0% FBS), cells were pre-incubated with dDAVP (1 pM), and then 0.7-1 x 10A5 cells were seeded into 8 pm-pore inserts (JetBiofil, Guangzhou, China) in serum-free DMEM or RPMI and dDAVP. Lower chambers were filled with 10% FBS-containing DMEM or RPMI as chemoattractant. After 17 h-incubation with dDAVP (1 pM), cells present in the upper surface of the membranes were removed with cotton swabs and cells that migrated through the insert and attached to its lower surface were fixed, stainedand washed. Migrating cells in 4-5 randomly selected X400-high power field (HPF) per insert were quantified using QuPath v0.3.0 and normalized to control.Performed by triplicate (independent operators), 8-10 X400-HPF per experimental group per independent replicate.

[0307] GBM log-phase proliferation

[0308] A high-density log-phase proliferation assay was used to evaluate GBM cellular growth. Tumour cell seeding was conducted at high density (2.5-4x10A3) in 100 pL of complete medium (96 multi-well plate format). After 24 h, cells are exposed to dDAVP (concentration range of 0.1-10 pM). 72 h later exponentially growing cell cultures were quantified by the metabolic MTS (Absorvance measurement at A=490 nm, Biotek synergy HT multimode reader) or fixed for 10 min with methanol and then stained with 0.5% crystal violet solution for another 10 min. Following a washing step, fixed cultures were dried overnight. Quantification for the crystal violet method was performed by absorbance measurement (A=595 nm) after adding an ethanol / acetic acid 3:1 solution. Values are presented as a percentage of control. Performed by duplicate (independent operators), 6-12 wells per experimental group per independent replicate. High resolution photographs were taken from different representative wells from each experimental group.

[0309] Combinational studies of dDAVP addition to TMZ

[0310] Clonogenic assays were conducted in order to evaluate the cytostatic effect of dDAVP in addition to TMZ on GBM, as previously described. As depicted in Figure 14, two different in vitro combinational treatment schemes were assessed, a sequential exposure to dDAVP alone (from day 0-3) and then to TMZ alone (from day 3-7), and a concomitant exposure to dDAVP in addition to TMZ (from day 3-7). All monotherapies were independently evaluated. Performed by duplicate (independent operators), 6 wells per experimental group per independent replicate.

[0311] A VPR2 expression assessment In GBM cells by RT-qPCR

[0312] Total RNA of LN229, A172, MG-63 (human AVPR2-expressing osteosarcoma, C+) and U-2 OS (human AVPR2-negative osteosarcoma, C-) cells was purified from 1A106cells using Bio-Zol (Productos Bio-L6gicos, Argentina) andwas reverse-transcribed with MMLV Transcripta (Productos Bio-L6gicos, Argentina), both according to the manufacturer’s protocol. The following forward and reverse primers were used: for AVPR2 5’-CTGGCCAAGGACACTTCATC-3’ (SEQ ID NO: 1) and 5’-GAAGGCAGCTGAGCTTC-3 (SEQ ID NO: 2)’, for p-actin (ACTB), 5'-GGA CTT CGA GCA AGA GAT GG-3' (SEQ ID NO: 3) and 5'-AGG AAG GAAGGC TGG AAGAG-3' (SEQ ID NO: 4). Quantitative real time polymerase chain reaction (RT-qPCR) was performed using the SYBR Green PCR Master Mix (Thermo Fisher Scientific Inc.) and StepOne Real-Time PCR System (Applied Biosystems, California, United States). Thermal cycling conditions were 48°Cfor30 min, 95°C for 10 min, 40 cycles of 95°C for 15 sec followed by 60°C for 60 sec. Each sample was analysed in triplicate and mean cycle threshold values (Ct) were used for further analysis. Ct values were normalized for HPRT 1 expression levels. The AVPR2 expression in MG-63 cells (C+, positive control) was considered as “100%”. Relative quantification values were calculated as 2(-AACt)(Sobol et al.).

[0313] Immunohistochemical staining for A VPR2 in GBM xenografts

[0314] LN229 GBM xenografts were generated in athymic nude mice by orthotopic surgical brain implantation, as reported (Cardama et al). After 90 days of tumour progression, tumours were resected and processed. LN229 GBM tumours were fixed in 10% formalin, embedded in paraffin, and tissue sections of 4 pm were cut and placed on silane coated slides for anti-AVPR2 immunohistochemistry (IHC) staining. Sections were microwaved for antigen retrieval with Tris-EDTA buffer (pH 9.0) during 20 min, and later incubated with 3% hydrogen peroxide for 10 min to block endogenous peroxidase. Immunostaining was performed using an anti-AVPR2 primary monoclonal rabbit antibody (Clone EPR24555-59; 1:50 dilution, Abeam) at room temperature for 1 h. For visualizing immunoreactivity, a 45-min incubation using PolyDetector DAB HRP Brown Immunohistochemistry (IHC) detection system (BioSB, California, United States) was conducted. Finally, sections were counterstained with hematoxylin.

[0315] Bioinformatic studies

[0316] In silico studies aiming at assessing AVPR2 expression in GBM and low grade glioma (LGG), its impact on disease progression and other relevant biologic and clinical markers, were conducted using the GEPIA (Gene Expression Profiling InteractiveAnalysis) V2.0 and the TIMER (Tumor Immune Estimation Resource) V2.0 platforms. GEPIA is an interactive web platform designed for bioinformatic analysis of cancer transcriptomics, specifically utilizing data from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) projects. For survival analysis Kaplan-Meier Curve Parameters: Split Expression Percentage of Patients 45 (%) and Survival Time Between 0-80 (Months). Additionally, the TIMER platform is a web-based tool designed for the comprehensive analysis of tumour-infiltrating immune cells, particularly utilizing data from The Cancer Genome Atlas (TCGA). TIMER 2.0, the latest version, integrates multiple computational algorithms to estimate the abundance of various immune cell types within tumour samples, aiding researchers in understanding the interplay between tumour biology and immune response. TCGA GBM n=153. TCGA LGG n=516.

[0317] Statistics

[0318] In vitro assay determinations were conducted from at least 2 or 3 independent experiments, with a variable number of replicates per experiment type ranging from 3-12 per experimental group. Statistical analyses were performed using GraphPad Prism 8.0V software, (GraphPad Prism Software, USA). Distribution of data and descriptive statistics were firstly assessed. Outliers were detected and removed (Prism calculator). For comparisons among 3 or more experimental groups, ANOVA or its non-parametric equivalent, the Kruskal-Wallis test, were utilized. Subsequently, tests such as Dunnett's test, Tukey's test, comparison of 95% confidence intervals (Cl) for the mean (ANOVA), or Dunn's test (Kruskal-Wallis) were employed. For comparisons between 2 groups, either the Student t-test or the Mann-Whitney test were used, depending on the parametric or nonparametricdistribution of values, respectively. For the calculation of the half and quarter inhibitory concentration (IC50 and IC25), linear and nonlinear regressions were applied, depending on optimal curve-fitting. Differences were considered statistically significant at a level of p<0.05. *p<0,05 ; **p<0,01 ; ***p<0,001 ;****p<0,0001

[0319] Results and discussion

[0320] After surgical resection and adjuvant therapies, residual GBM cells can survive and proliferate, leading to the establishment and outgrowth of tumour colonies and recurrence (Babaloui et al.).

[0321] We first evaluated dDAVP effect on GBM colony formation and outgrowth using a long term clonogenic assay. After one week of drug exposure, dDAVP, even at low micromolar concentrations, was able to significantly reduce clonogenic growth, impairing tumour colony formation and progression by up to =60% at 1 pM. An IC50 of 0.8 pM wasobtained of long-term low-density GBM cultures (Figure 1). We further determined the direct inhibitory effect on A172 long-term cultures by dDAVP at 1 pM obtaining an equivalent (estimated) IC50 value of 0.7 pM. (Figure 2).

[0322] These findings suggest that dDAVP administration in the perisurgical window may limit the survival of residual cells and reduce their ability to consolidate a recurrent lesion.

[0323] GBM is characterized by its highly infiltrative nature, which is one of the key features that contribute to its lethality and poor prognosis. This infiltrative phenotype allows tumour cells to invade surrounding brain tissue extensively, making complete surgical resection challenging and often ineffective in preventing recurrence. GBM uses mesenchymal migration to invade and spread. This mode of migration is a significant clinical challenge (Zhong et al., Rezk et al.).

[0324] We determined the impact of dDAVP on the phenomenon of tumour chemotaxis of GBM cells.

[0325] dDAVP displayed a significant inhibitory activity on tumour cell motility of A172 cells, reducing chemotactic capacity by 14% in treated cells (Figure 3). It is worth noting that this GBM cell line displays increased mesenchymal features and it is widely recognized as a highly migratory and invasive model. In contrast, a non-significant 8% reduction in GBM chemotaxis was observed after treatment in less migratory and epithelial-type LN229 cells (Figure 4). As a result, a significant inhibition of the migratory ability of remnant highly-motile malignant cells after tumour resection may limit their dissemination and spread, helping control local disease.

[0326] High proliferation rates in GBM, are biologically significant as they directly correlate with tumour aggressiveness, growth dynamics and recurrence, and patient prognosis (Stensj0en et al.). Untreated GBM established lesions follow exponential growth patterns with volume duplicating time as low as a month (Feucht et al.).

[0327] It was further evaluated if dDAVP was able to impair the proliferation of GBM in log-phase growing tumour cell cultures. In these experiments, dDAVP displayed a direct antiproliferative effect on rapidly-growing LN229 cell cultures, obtaining a IC25 value of 1.22 pM. A significant antimitotic action was observed even at clinically-relevant low drug concentrations of 100 nM (0.1 pM)(Figure 5). A significant, but less potent antiproliferative effect of dDAVP was observed on high-density mesenchymal-type A172 cultures, with an increased IC25 value of 11.8 pM (Figure 6).

[0328] dDAVP antiproliferative action may stall the progression of rapidly-growing recurrent GBM lesions and may complement and enhance the activity of standard-of-care chemotherapy, such as TMZ.

[0329] The molecular target expression in GBM models, both in vitro and in vivo, was determined.

[0330] First, AVPR2 gene expression by GBM cells was determined by qRT-PCR. LN229 and A172 cells express equivalent levels of AVPR2 mRNA cells in comparison to C+ target-expressing human osteosarcoma MG-63 cells (mesenchymal tumour). LN229, A172 and MG-63 (C+) express significant higher levels of AVPR2 versus U2-OS cells, a well established negative model for AVPR2 expression which do not respond in vitro to dDAVP stimulation (Figure 7).

[0331] In addition, we assessed AVPR2 expression in vivo by IHC in LN229 human GBM xenografts generated in FoxN1nu / nuathymic nude mice.

[0332] GBM xenografts clearly displayed inmunoreactivity against AVPR2, with high heterogeneity between tumour regions, going from weak and “patchy” patterns of positivity to strong focal immunoreactivity (Figure 8). In figure 8 different patterns of immunoreactivity for AVPR2 are depicted, in addition to representative microphotographs positive and negative expression controls: (A) LN229 anti-AVPR2 strong focal immunoreactivity; (B) LN229 anti-AVPR2 moderate immunoreactivity; (C) LN229 anti-AVPR2 weak and “patchy” immunoreactivity; (D) LN229 anti-AVPR2 weak / moderate focal immunoreactivity; (E) C+ (MG-63 human osteosarcoma xenograft, moderate immunoreactivity); and (F) C- (Omission of 1ary monoclonal Antibody anti-AVPR2). GBM xenografts displayed inmunoreactivity against AVPR2, with high heterogeneity between tumour regions, going from weak and “patchy” patterns of positivity to strong focal immunoreactivity. MG-63 human osteosarcoma xenograft samples were used as AVPR2-positive controls.

[0333] Once target expression was determined in GBM models, we evaluated AVPR2 impact on GBM progression and its correlation with different clinical biomarkers. Using TIMER bioinformatic platform and TOGA database we found a significant favourable impact of high AVPR2 expression on overall survival in GBM patients, after combining all molecular subtypes and adjusting by secondary clinical factors such as gender, race and age (Figure 9). Moreover, using GEPIA platform and TOGA plus GTEx transcriptomics databases we observed a lower (strong tendency, but non significant) AVPR2 expressionin GBM tumour samples in comparison to normal matched tissue (Figure 10). Same expression profile was observed for LGG clinical samples versus normal tissue.

[0334] The infiltration of immune cells, particularly CD4+ T cells, CD8+ T cells, B cells and natural killer (NK) cells, plays a crucial role in the biological behavior of GBM and their response to therapies (Han et al). Using the TIMER platform (which allows through deconvolution methods on RNAseq data to estimate the degree of infiltration of different immune populations in the TME) correlations between AVPR2 expression and key immune infiltrates in the GBM stroma were assessed.

[0335] First, in GBM, as well as in LGG, AVPR2 was maintained despite changes in tumour purity. Interestingly, target expression correlates positively with higher infiltration of CD8+ cytotoxic T cells, NK and B cells in the TME, all immune populations which are linked to antitumour responses (Figure 11 A and 11 B). Equivalent results were obtained in LGG, after observing correlation coefficients > 0.22-0.26, p values < 0.05 for CD4+ and CD8+ T cell populations, NK and B cell infiltration (Figure 12). High AVPR2 expression was associated with increased antitumoural immune infiltration in the tumour stroma of low and high grade gliomas, and could be correlated with a more immunoreactive TME that may be leveraged to increase response to chemotherapy or even immunotherapy.

[0336] TMZ-based chemotherapy is widely used in the treatment of GBM, but it has several limitations that affect its efficacy and patient outcomes, such as resistance development by tumour cells (including MGMT overexpression), serious adverse effects related to treatment, limited and uneven penetration in the brain, and lack of synergistic effects with other cytotoxic agents (Oraiopoulou et al., Chamberlain). In order to assess potential therapeutic benefits after combining dDAVP with TMZ we first evaluated the TMZ effect, as a monotherapy, on long-term GBM LN229 cultures. TMZ displayed a concentration-dependent inhibitory effect on GBM long-term growth with an IC50 value of 10,7 pM after a 7 d-drug exposure (Figure 13). Once we characterized the cytotoxic effect of TMZ on LN229 cells, two different treatment schemes for dDAVP plus TMZ combinational studies were determined. Different experimental designs under assessment are depicted in Figure 14, showing a sequential (pre-treatment with dDAVP alone for the first 3 d, and then exposure to TMZ alone for the remaining 4 d) or concomitant (dDAVP co-treatment in addition to TMZ, from day 4 onwards) therapeutic schemes (groups number 5 or 6, respectively) in the dual therapy groups.

[0337] Surprisingly, a cooperative inhibitory effect was observed in sequential combined exposure to dDAVP and then TMZ, enhancing therapeutic benefits incomparison to each monotherapy alone. Concomitant use of dDAVP in addition to TMZ from day 4 onwards does not improve the anti-GBM activity versus respective monotherapies. Regarding the effects of different monotherapies, it is with noting that early exposure to dDAVP seems to be quite determinant in its inhibitory effect on GBM clonogenic growth (equivalent effect to full 7 d exposure). However, late exposure to dDAVP still significantly impairs clonogenic growth, but to a lesser extent. In addition, late 4 d-exposure to TMZ had a similar effect to 7 d exposure.

[0338] Discussion

[0339] These findings establish for the first time that desmopressin exhibits an antitumour effect in models of human GBM, utilizing well-established cell lines to explore new therapeutic strategies for combating this aggressive form of brain cancer.

[0340] Surprisingly, the combination of dDAVP and TMZ was effective when cancer cells are sequentially exposed first to dDAVP and then to TMZ, rather than when the compounds are applied concomitantly within the same time window.

[0341] These results also indicate that dDAVP may be applied as a neoadjuvant treatment during the perioperative period, and / or in the immediate postoperative phase, before cycles of TMZ-based chemotherapy.

[0342] References

[0343] Alves ALV, et al. Role of glioblastoma stem cells in cancer therapeutic resistance: a perspective on antineoplastic agents from natural sources and chemical derivatives. Stem Cell Res Ther. 2021 Mar 24;12(1):206.

[0344] Babaloui S, et al. Radiosensitization of Glioma Cells by Temozolomide (TMZ): A Colony Formation Assay. J Biomed Phys Eng. 2022 Feb 1 ;12(1):43-50.

[0345] Berte N, et al. Artesunate enhances the therapeutic response of glioma cells to temozolomide by inhibition of homologous recombination and senescence. Oncotarget.2016 Oct 11;7(41):67235-67250.

[0346] Birzu C, et al. Recurrent Glioblastoma: From Molecular Landscape to New Treatment Perspectives. Cancers (Basel). 2020 Dec 26;13(1):47.

[0347] Cardama GA, et al. Preclinical Efficacy and Toxicology Evaluation of RAC1 Inhibitor 1A-116 in Human Glioblastoma Models. Cancers (Basel). 2022 Sep 30;14(19):4810.

[0348] Celiku O, et al. Computational modeling demonstrates that glioblastoma cells can survive spatial environmental challenges through exploratory adaptation. Nat Commun. 2019 Dec 13;10(1):5704.

[0349] Chamberlain MC. Temozolomide: therapeutic limitations in the treatment of adult high-grade gliomas. Expert Rev Neurother. 2010 Oct; 10(10): 1537-44.

[0350] Daniel PM, et al. Sensitivity of GBM cells to cAMP agonist-mediated apoptosis correlates with CD44 expression and agonist resistance with MAPK signaling. Cell Death Dis. 2016 Dec 1 ;7(12):e2494.

[0351] Feucht D, et al. Preoperative growth dynamics of untreated glioblastoma: Description of an exponential growth type, correlating factors, and association with postoperative survival. Neurooncol Adv. 2024 Apr 3;6(1):vdae053.

[0352] Jha RM et al. Emerging therapeutic targets for cerebral edema. Expert Opin Ther Targets. 2021 Nov;25(11):917-938.

[0353] Karmur BS et al. Blood-brain barrier disruption in Neuro-Oncology: strategies, failures, and challenges to overcome. Front Oncol 10: 563840, 2020.

[0354] Mohammed S, et al. Survival and quality of life analysis in glioblastoma multiforme with adjuvant chemoradiotherapy: a retrospective study. Rep Pract Oncol Radiother. 2022 Dec 29;27(6): 1026-1036.

[0355] Morales DE, Mousa S. Intranasal delivery in glioblastoma treatment: prospective molecular treatment modalities. Heliyon 8: e09517, 2022.

[0356] Oraiopoulou ME,et al. The Temozolomide-Doxorubicin paradox in Glioblastoma in vitro-in silico preclinical drug-screening. Sci Rep. 2024 Feb 14;14(1):3759.

[0357] Rezk R, et al. Spatial heterogeneity of cell-matrix adhesive forces predicts human glioblastoma migration. Neurooncol Adv. 2020 Jul 3;2(1):vdaa081.

[0358] Sharma P, et al. Tumor microenvironment in glioblastoma: Current and emerging concepts. Neurooncol Adv. 2023 Feb 23;5(1):vdad009.

[0359] Shi L, et al. The DRD2 Antagonist Haloperidol Mediates Autophagy-Induced Ferroptosis to Increase Temozolomide Sensitivity by Promoting Endoplasmic Reticulum Stress in Glioblastoma. Clin Cancer Res. 2023 Aug 15;29(16):3172-3188.

[0360] Siegner H et al. Does DDAVP (1-Desamino-8-D- Arginine-Vasopressin) Cross the Blood-CSF Barrier? Neuroendocrinology 37: 262-265, 1983.

[0361] Sobol NT, et al. Anticancer activity of repurposed hemostatic agent desmopressin on AVPR2-expressing human osteosarcoma. Exp Ther Med. 2021 Jun;21(6):566.

[0362] Sorensen PS et al. Impermeability of the blood-cerebrospinal fluid barrier to 1-deamino-8-D-arginine-vasopressin (DDAVP) in patients with acquired, communicating hydrocephalus. European Journal of Clinical Investigation 14: 435-439, 1984.

[0363] Stensj0en AL, Solheim O, Kvistad KA, Haberg AK, Salvesen 0, Berntsen EM. Growth dynamics of untreated glioblastomas in vivo. Neuro Oncol. 2015 Oct; 17(10):1402-11.

[0364] Sugimoto N, et al. Targeted activation of PKA and Epac promotes glioblastoma regression in vitro. Mol Clin Oncol. 2013 Mar;1(2):281-285.

[0365] Vollmann-Zwerenz A, Leidgens V, Feliciello G, Klein CA, Hau P. Tumor Cell Invasion in Glioblastoma. Int J Mol Sci. 2020 Mar 12;21(6):1932.

[0366] Xing F, et al. The Anti-Warburg Effect Elicited by the cAMP-PGC1a Pathway Drives Differentiation of Glioblastoma Cells into Astrocytes. Cell Rep. 2017 Jan 10;18(2):468-481. doi: 10.1016 / j.celrep.2016.12.037. Erratum in: Cell Rep. 2018 May 29;23(9):2832-2833.

[0367] Zhong J, et al. Mesenchymal migration as a therapeutic target in glioblastoma. J Oncol. 2010;2010:430142.

[0368] The embodiments described herein are intended to be examples only. Alterations, modifications and variations can be effected to the particular embodiments by those of skill in the art. The scope of the claims should not be limited by the particular embodiments set forth herein, but should be construed in a manner consistent with the specification as a whole.

[0369] EXAMPLE 2

[0370] Materials and methods

[0371] Drugs and reagents

[0372] Desmopressin acetate (dDAVP; MW 1183.2 g / mol) was obtained from Ferring, in injectable form. Subsequent dilutions using saline were carried out to attain final working concentrations for in vivo studies. Temozolomide (TMZ; MW 194.151 g / mol) was obtained from Sigma Aldrich, in powder form. The compound was initially dissolved in DMSO to generate concentrated stocks. Subsequent dilutions using DMSO were carried out to attain final working concentrations for in vivo studies. PBS or appropriate concentrations of DMSO, respectively, were used as vehicles for the control groups.

[0373] Cell lines and culture conditions

[0374] LN229 (ATCC CRL-2611 ™) is a highly aggressive human GBM cell model, widely used in biomedical research. It was isolated in 1979 from the right frontalparieto-occipital cortex of a White, 60-year-old, female patient with GBM. It has an epithelial-like morphology and it is tumourigenic in nude mice. It bears a mutation in p53 (TP53) and possible homozygous deletions in the p16 and p14ARF tumour suppressor genes. LN229 cells were grown using Dulbecco's modified Eagle's medium (DMEM) culture medium (Gibco, USA) and Roswell Park Memorial Institute medium (RPMI), respectively, with 10% FBS (Natocor, Argentina), 80 pg / ml gentamicin, and 2mM glutamine, in monolayer culture, at 37°C in a humidified atmosphere of 5% CO2. Cells were harvested using a trypsin / EDTA solution (Thermo Fisher Scientific Inc., USA) diluted in PBS and routinely tested for mycoplasma.

[0375] Animals

[0376] Inbred athymic female and male N:NIH(S)-nu mice aged 6 weeks with a weight of approximately 23 g, were purchased from the School of Veterinary Sciences Animal Facility at National University of La Plata (Buenos Aires, Argentina). After randomization, animals were housed at 5 mice per cage in our animal facility at the National University of Quilmes. Food and water was provided ad libitum and the general health status of the animals was monitored daily. Animal enrichment elements were used routinely according to international IACUC recommendations (Physical Habitat, Sensory and Food Enrichment). 5 animals were used per experimental group. All in vivo protocols were revised and approved by our institutional animal care and use committee (IACUC; UNQ-DCyT-CICUAL Res.CD N°129 / 17; CD N°016 / 24; Protocol code N°009-25).

[0377] GBM xenograft generation

[0378] Human GBM tumours were heterotopically generated after subcutaneous injection of a 250 pl suspension containing 7 x 106LN229 cells in DMEM and Matrigel (Corning, New Jersey, USA) in a 1.5:1 volume ratio in athymic mice (Wang et al.; Jian et al.; Wang (II) et al.). Matrigel is a commercially available extracellular matrix extract used for tumour engraftment in xenogeneic models, favoring tumour establishment and outgrowth. Tumour formation was assessed daily by palpation. Once confirmed, tumours were measured periodically (two times per week) with a caliper and tumour volume was calculated by the formula: 0.52 x width2x length. During the protocol animal weights and tumour growth rates (TGR) were also assessed. Photographic records of the GBM-bearing mice were taken along the protocols. Animals were sacrificed when GBM xenografts crossed the human cut-off value of tumour volume >1000 mm3.

[0379] Treatment schemes

[0380] Preclinical protocol design: Administration of dDAVP in the peri-tumour implantation window, alone or followed by adjuvant TMZ (Figure 17)

[0381] First, animals were randomized in 4 independent experimental groups; Control, dDAVP, TMZ and dDAVP+TMZ, housed at 5 animals per cage. dDAVP was used at a dose of 4 pg / kg i.v. and was administered peri-implantation of tumour cells (-30 minutes and +24 hours), and in two weekly boosters on days 7 and 14 of the protocol (dDAVP and dDAVP+TMZ groups). dDAVP dosing in experimental animals was determined by dose extrapolation using the ‘dose by factor’ method based on allometric scaling, following the United States Food and Drug Administration guidelines (Nair et al.). A clinically validated dDAVP dose of 0.3 pg / kg i.v. was taken as the reference dose and multiplied by 12.3 for human to mice conversion. Mice equivalent dose was rounded up from 3.7 to 4 pg / kg i.v. to facilitate drug preparation and administration. During the 3rd week, >50% of animals belonging to the control reference group were positive for GBM engraftment, meeting the criteria for starting TMZ-based chemotherapy, and stopping dDAVP administration in the corresponding groups. Consequently, treatment with TMZ (in the TMZ and dDAVP+TMZ groups) began on day 21 , on a weekly basis, with a suboptimal dose of 10 mg / kg i.p. The incidence of tumours was 100% for all groups at day 26. Prior to the completion of the protocol and the sacrifice of the animals at day 58 the mice belonging to the groups receiving chemotherapy were administered with 6 cycles of TMZ in total, following a clinical-translational criterion.

[0382] Preclinical protocol design: Co-adjuvant use of dDAVP with concomitant TMZ-based chemotherapy (Figure 18)

[0383] Animals were randomized in 3 independent experimental groups; Control, TMZ and dDAVP+TMZ, and housed at 5 animals per cage.

[0384] At day 15, >50% of animals were positive for GBM engraftment, meeting the criteria for starting TMZ or TMZ+dDAVP-based therapy. The incidence of tumours was 100% for all groups at day 19. Treatment with dDAVP (4 pg / kg i.v.) was only administered prior (-30 to -60 minutes) to TMZ. Consequently, treatment with TMZ (in the TMZ and dDAVP+TMZ groups) began on day 15, on a weekly basis, with a suboptimal dose of 10 mg / kg i.p. Prior to the completion of the protocol and the sacrifice of the animals at day 51 , the mice belonging to the groups receiving chemotherapy were administered with 6 cycles of TMZ in total, once again, following a clinical-translational criterion.

[0385] Statistics

[0386] Statistical analyses were performed using GraphPad Prism 8.0V software, (GraphPad Prism Software, USA). Distribution of data and descriptive statistics were firstly assessed. Outliers were detected and removed (Prism calculator). For comparisons among 3 or more experimental groups, ANOVAor its non-parametric equivalent, the Kruskal-Wallis test, were utilized. Subsequently, tests such as Dunnett's test, Tukey's test, comparison of 95% confidence intervals (Cl) for the mean (ANOVA), or Dunn's test (Kruskal-Wallis) were employed. For the calculation of the TGR linear regressions were applied. Impact of dDAVP on tumour engraftment was assessed using the chi-square statistical test at day 18. Differences were considered statistically significant at a level of p<0.05. *p<0,05 ; **p<0,01 ; ***p<0,001 ;****p<0,0001

[0387] Results

[0388] Preclinical protocol design: Administration of dDAVP in the peri-tumour implantation window, alone or followed by adjuvant TMZ

[0389] After surgical resection residual GBM cells can survive and proliferate, leading to the rapid establishment and outgrowth of aggressive tumour implants and recurrence (Babaloui et al.; Stensj0en et al.). Untreated GBM established lesions follow exponential growth patterns with volume duplicating time as low as a month (Feucht et al.). Taking that into consideration, we first aimed at evaluating the impact of early dDAVP administration on human GBM xenograft establishment and its subsequent progression in nude mice. In this setting, dDAVP was administered peri-implantation of GBM cells and in two weekly boosters, as a monotherapy or before starting TMZ cytotoxic therapy. Table 1 depicts days of confirmed tumour engraftment in different experimental groups. Mean latency for control, dDAVP, TMZ and dDAVP plus TMZ groups was 15.6±3.4, 19.4±5.5, 16.4±6.8 and 18.2±5.5 days, respectively. Results show that dDAVP administration prior and immediately after tumour cell injection adversely impacts tumour engraftment and early progression of GBM in vivo. This trend was observed in the dDAVP-treated groups (dDAVP and dDAVP+TMZ), in contrast to the reference groups (control and TMZ), as observed in Tables 1 and 2. Specifically, on day 18, engraftment was 100% for the control group and 40% for the dDAVP monotherapy group, reaching statistical significance (dDAVP versus Control, Chi-square: 4.286, p-value*: 0.038). At day 26 tumour take was confirmed for all animals in all 4 experimental groups (Table 2).Latency (day of tumor engraftment)

[0390] Table 1. GBM tumour latency expressed as days until xenograft engraftment. *At day 18, dDAVP versus Control, Chi-square: 4.286, p-value: 0.038„ Incidence at day 26 LatencyTreatment (Pos .it..ive / >Tot .al) (Days±SD)Control 5 / 5 15,6±3,6dDAVP 5 / 5 19,4±5,5TMZ 5 / 5 16,4±6,8dDAVP+TMZ 5 / 5 18,2±5,5

[0391] Table 2. Incidence of GBM xenografts at day 26 and mean latency for each experimental group.

[0392] Progression of mean tumour volume of each group over time and GBM volume analysis at fixed days are depicted in Figure 19A or 19B (Day 36), 19C (Day 43), 19D (Day 51) or 19E (Day 58). As observed in Figure 19A, established tumours belonging to control vehicle-treated mice grew following an exponential pattern, confirming a highly aggressive disease. In contrast, although with varying degrees of therapeutic outcome, all therapies managed to modulate the aggressiveness of GBM xenotransplants. First, at day 36, all treatments deploy equivalent anti-GBM effects. Despite the fact that sustained TMZ, given as a monotherapy, was capable of stabilizing GBM growth, throughout the rest of the preclinical trial, animals receiving dDAVP in addition to TMZ-based therapy showed the smallest recorded xenotransplants in average (Day 40 onwards). Although at a less noticeable level than TMZ-based therapies, the tumours initially treated with dDAVP progressed more slowly than the tumours in the control group.

[0393] Individual or average tumour growth rates (between days 19 and 51 of the protocol) of GBM xenografts growing in female nude athymic mice are shown in Figure 20A or 20B, respectively. As observed in the individual analysis, the response to treatment and the consequent modulation of progression rates show a high level of clustering and consistency in the impact of the different therapies (Figure 20A). Mean tumour growth rates for control, dDAVP, TMZ, or dDAVP plus adjuvant chemotherapywere found as 34.7, 22.8, 0.5 or -1,9 mm3 / day. All evaluated therapies negatively modulated GBM growth dynamics when compared to the progression rates of the rapidly growing control tumours. The group initially treated with dDVAP plus sustained TMZ performed significantly better than dDAVP or TMZ alone, being the only approach associated with disease regression (negative tumour growth rates). Surprisingly, dDAVP monotherapy administered only in the early window of tumour establishment was sufficient to significantly alter and inhibit long-term GBM progression in comparison to vehicle-treated animals.

[0394] Representative lesions in GBM-bearing mice from different experimental groups are depicted in Figure 21.

[0395] Regarding the safety of the therapies evaluated, all treatments were well tolerated. No significant changes were observed in behavior, water and food consumption, or animal weight in relation to any of the treatments evaluated. An upward trend in animal weight was observed in all experimental groups, which is consistent with the young age of the experimental subjects, as well as with the relatively good tolerance of the monotherapies or combination therapies (Figure 22A and 22B). After analyzing the relative weight gain of animals on day 0 of the protocol, it is worth noting that at the midpoint of the protocol (day 35), the group with the greatest weight gain was the one receiving combined dDAVP plus chemotherapy, even after 3 cycles of TMZ, showing an increase of 26% in body weight. Relative animal weight gain associated with dDAVP, TMZ or vehicle treatment was found as 24, 17 and 18%, respectively (Figure 22B, nonsignificant, p>0.05).

[0396] Preclinical protocol design: Co-adjuvant use of dDAVP with concomitant TMZ-based chemotherapy

[0397] In this second preclinical trial, once GBM disease was confirmed, coadjuvant dDAVP was used at a dose of 4 pg / kg i.v. and was given immediately prior to weekly TMZ cycles (10 mg / kg i.p.). TMZ was also assessed as a monotherapy. At day 19 tumour take was confirmed for all animals in all 3 experimental groups and equivalent tumour volumes were recorded for all animals.

[0398] Progression of mean tumour volume of each group over time and GBM volume analysis at fixed days are depicted in Figure 23A or 23B (Day 26), 23C (Day 36), 23D (Day 40) or 23E (Day 51). As previously reported and as observed in Figure 23A, xenografts belonging to vehicle-treated control animals grew rapidly, confirming a highly proliferative and aggressive GBM model in vivo. More than two weeks of chemotherapyexposure are required to visualize the effect of TMZ as monotherapy (Figure 23A).Surprisingly, the addition of dDAVP to TMZ enhances the activity of chemotherapy alone, allowing for rapid arrest of GBM growth and progression in the early stages of the disease. The potent long-term effect of TMZ, both alone and in combination with dDAVP, is noteworthy.

[0399] Both therapies negatively modulate GBM growth dynamics when compared to the progression rate of the control group. The group treated with dDAVP and TMZ performed significantly better than TMZ alone, being the only therapeutic approach associated with disease regression (negative growth slope in the linear regression analysis). Mean tumour growth rates for control, TMZ, or adjuvant dDAVP plus TMZ were found as 22.3, 0.8 or -1 ,2 mm3 / day (Figure 24B).

[0400] Representative lesions in GBM-bearing mice from control, TMZ or dDAVP plus TMZ experimental groups are depicted in Figure 25.

[0401] Regarding toxicity, both therapies were well tolerated. No significant changes were observed in behavior, water and food consumption, or animal weight in relation to any of the treatments evaluated. Once again, an upward trend in animal weight was observed in all experimental groups, which is consistent with the young age of the experimental subjects, as well as with the relatively good tolerance of the monotherapies or combination therapies (Figure 26A and B).

[0402] Discussion

[0403] These in vivo findings show for the first time that dDAVP exhibits an antitumour effect in animal models of human GBM, utilizing a well-established and aggressive cellular model to explore new therapeutic strategies for combating this deadly form of brain cancer.

[0404] dDAVP, using clinically relevant doses, was tested as a neoadjuvant treatment during tumour cell injection, with weekly additional doses in the post-tumour challenge window, resembling the use in the perioperative period and immediate postoperative phase, before cycles of TMZ-based chemotherapy.

[0405] Secondly, sustained dDAVP was administered weekly upon tumour establishment, prior to GBM-specific cytotoxic treatment. In both tested experimental scenarios, dDAVP was associated with therapeutic benefits on latency of implanted GBM cells and / or progression rate of established disease, especially in addition to standard chemo.

[0406] References (alphabetical order)

[0407] Alves ALV, et al. Role of glioblastoma stem cells in cancer therapeutic resistance: a perspective on antineoplastic agents from natural sources and chemical derivatives. Stem Cell Res Ther. 2021 Mar 24;12(1):206.

[0408] Babaloui S, et al. Radiosensitization of Glioma Cells by Temozolomide (TMZ): A Colony Formation Assay. J Biomed Phys Eng. 2022 Feb 1 ;12(1):43-50.

[0409] Birzu C, et al. Recurrent Glioblastoma: From Molecular Landscape to New Treatment Perspectives. Cancers (Basel). 2020 Dec 26;13(1):47.

[0410] Cardama GA, et al. Preclinical Efficacy and Toxicology Evaluation of RAC1 Inhibitor 1A-116 in Human Glioblastoma Models. Cancers (Basel). 2022 Sep 30;14(19):4810.

[0411] Celiku O, et al. Computational modeling demonstrates that glioblastoma cells can survive spatial environmental challenges through exploratory adaptation. Nat Commun. 2019 Dec 13;10(1):5704.

[0412] Chamberlain MC. Temozolomide: therapeutic limitations in the treatment of adult high-grade gliomas. Expert Rev Neurother. 2010 Oct; 10(10): 1537-44.

[0413] Daniel PM, et al. Sensitivity of GBM cells to cAMP agonist-mediated apoptosis correlates with CD44 expression and agonist resistance with MAPK signaling. Cell Death Dis. 2016 Dec 1 ;7(12):e2494.

[0414] Feucht D, et al. Preoperative growth dynamics of untreated glioblastoma: Description of an exponential growth type, correlating factors, and association with postoperative survival. Neurooncol Adv. 2024 Apr 3;6(1):vdae053.

[0415] Garona J (II), et al. Addition of vasopressin synthetic analogue [V(4)Q(5)]dDAVP to standard chemotherapy enhances tumour growth inhibition and impairs metastatic spread in aggressive breast tumour models. Clin Exp Metastasis. 2016 Aug;33(6):589-600.

[0416] Garona J, et al. The novel desmopressin analogue [V4Q5]dDAVP inhibits angiogenesis, tumour growth and metastases in vasopressin type 2 receptor-expressing breast cancer models. Int J Oncol. 2015;46(6):2335-45.

[0417] Jha RM et al. Emerging therapeutic targets for cerebral edema. Expert Opin Ther Targets. 2021 Nov;25(11):917-938.

[0418] Jiang S, et al. Double-modified oncolytic adenovirus armed with a recombinant interferon-like gene enhanced abscopal effects against malignant glioma. Neurooncol Adv. 2023 Sep 28;5(1):vdad117.

[0419] Jones LW, et al. Effects of exercise training on antitumor efficacy of doxorubicin in MDA-MB-231 breast cancer xenografts. Clin Cancer Res. 2005 Sep 15;11(18):6695-8.

[0420] Karmur BS et al. Blood-brain barrier disruption in Neuro-Oncology: strategies, failures, and challenges to overcome. Front Oncol 10: 563840, 2020.

[0421] Mohammed S, et al. Survival and quality of life analysis in glioblastoma multiforme with adjuvant chemoradiotherapy: a retrospective study. Rep Pract Oncol Radiother. 2022 Dec 29;27(6): 1026-1036.

[0422] Morales DE, Mousa S. Intranasal delivery in glioblastoma treatment: prospective molecular treatment modalities. Heliyon 8: e09517, 2022.

[0423] Nair AB, Jacob S. A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm. 2016 Mar;7(2):27-31.

[0424] Oraiopoulou ME,et al. The Temozolomide-Doxorubicin paradox in Glioblastoma in vitro-in silico preclinical drug-screening. Sci Rep. 2024 Feb 14;14(1):3759.

[0425] Rezk R, et al. Spatial heterogeneity of cell-matrix adhesive forces predicts human glioblastoma migration. Neurooncol Adv. 2020 Jul 3;2(1):vdaa081.

[0426] Sharma P, et al. Tumor microenvironment in glioblastoma: Current and emerging concepts. Neurooncol Adv. 2023 Feb 23;5(1):vdad009.

[0427] Shi L, et al. The DRD2 Antagonist Haloperidol Mediates Autophagy-Induced Ferroptosis to Increase Temozolomide Sensitivity by Promoting Endoplasmic Reticulum Stress in Glioblastoma. Clin Cancer Res. 2023 Aug 15;29(16):3172-3188.

[0428] Siegner H et al. Does DDAVP (1-Desamino-8-D- Arginine- Vasopressin) Cross the Blood-CSF Barrier? Neuroendocrinology 37: 262-265, 1983.

[0429] Sobol NT, et al. Anticancer activity of repurposed hemostatic agent desmopressin on AVPR2-expressing human osteosarcoma. Exp Ther Med. 2021 Jun;21(6):566.

[0430] Sorensen PS et al. Impermeability of the blood-cerebrospinal fluid barrier to 1-deamino-8-D-arginine-vasopressin (DDAVP) in patients with acquired, communicating hydrocephalus. European Journal of Clinical Investigation 14: 435-439, 1984.

[0431] Stensj0en AL, Solheim O, Kvistad KA, Haberg AK, Salvesen 0, Berntsen EM. Growth dynamics of untreated glioblastomas in vivo. Neuro Oncol. 2015Oct; 17(10):1402-11.

[0432] Sugimoto N, et al. Targeted activation of PKA and Epac promotes glioblastoma regression in vitro. Mol Clin Oncol. 2013 Mar;1(2):281-285.

[0433] Vollmann-Zwerenz A, Leidgens V, Feliciello G, Klein CA, Hau P. Tumor Cell Invasion in Glioblastoma. Int J Mol Sci. 2020 Mar 12;21(6):1932.

[0434] Wang Y (II), et al. Zinc finger and SCAN domain-containing 18 suppresses the proliferation, self-renewal, and drug resistance of glioblastoma cells. Heliyon. 2023 Jun 3;9(6):e17000.

[0435] Wang Y, et al. Phenformin and metformin inhibit growth and migration of LN229 glioma cells in vitro and in vivo. Onco Targets Ther. 2018;11:6039-6048.

[0436] Xing F, et al. The Anti-Warburg Effect Elicited by the cAMP-PGC1a Pathway Drives Differentiation of Glioblastoma Cells into Astrocytes. Cell Rep. 2017 Jan 10;18(2):468-481.

[0437] All publications, patents and patent applications mentioned in this Specification are indicative of the level of skill those skilled in the art to which this invention pertains and are herein incorporated by reference to the same extent as if each individual publication patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0438] The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modification as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

Claims

WHAT IS CLAIMED:

1. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising, or consisting of: administering a therapeutically effective amount of desmopressin.

2. The method of claim 1 , wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

3. The method of claim 1 or 2, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

4. The method of any one of claims 1 to 3, wherein desmopressin is administered before, during, and / or after surgery to remove the glioblastoma.

5. The method of any one of claims 1 to 4, wherein desmopressin is administered perioperatively.

6. The method of any one of claims 1 to 5, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

7. The method of any one of claims 1 to 6, further comprising administering a therapeutically effective amount of temozolomide.

8. The method of claim 7, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

9. The method of any one of claims 7 to 8, wherein desmopressin is first administered to the subject followed by temozolomide administration to the subject.

10. The method of claim 9, wherein temozolomide is administered from about 3 days to about 7 days following desmopressin administration, from about 2 days to about 10 days following desmopressin administration, from about 1 day to about 12 days following desmopressin administration, from about 30 minutes to about 60 minutes following desmopressin administration.

11. The method of any one of claims 1 - 10, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

12. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising, or consisting of:- administering desmopressin prior to surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery;- administering desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma.13 The method of claim 12, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg14. The method of claim 12 or 13, further comprising administering temozolomide to said subject.

15. The method of claim 14, where said temozolomide is administered at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy.

16. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising or consisting of: administering a therapeutically effective amount of a vasopressin analogue.17 The method of any one of claim 16, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

18. The method of claim 16 or 17 wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

19. The method of any one of claim 16 to 18, wherein vasopressin analogue is administered before, during, and / or after surgery to remove the glioblastoma.

20. The method of any one of claims 16 to 19, wherein the vasopressin analogue is administered perioperatively.

21. The method of any one of claims 16 to 20, wherein the vasopressin analogue is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

22. The method of any one of claims 16 to 21, further comprising administering a therapeutically effective amount of temozolomide.

23. The method of claim 22, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

24. The method of any one of claims 22 to 23, wherein the vasopressin analogue is first administered to the subject followed by temozolomide administration to the subject.

25. The method of any one of claims 22 to 24, wherein temozolomide is administered from about 3 days to about 7 days following the vasopressin analogue administration, from about 2 days to about 10 days following the vasopressin analogue administration, from about 1 days to about 12 days following the vasopressin analogue administration, from about 30 minutes to about 60 minutes following the vasopressin analogue administration.

26. The method of any one of claims 16 -25, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

27. The method of any one of claims 16 to 26, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

28. The method of any one of claims 1 to 27, wherein the subject is a human.

29. A method for treating or preventing glioblastoma in a subject, comprising or consisting of:measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma ;and administering to a subject in need thereof a therapeutically effective amount of desmopressin, when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

30. The method of claim 29, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

31. The method of claim 30, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

32. The method of any one of claims 29 or 31 , wherein desmopressin is administered before, during, and / or after surgery to remove the glioblastoma.

33. The method of any one of claims 29 to 32, wherein desmopressin is administered perioperatively.

34. The method of any one of claims 29 to 33, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

35. The method of any one of claims 29 to 34, further comprising administering a therapeutically effective amount of temozolomide.

36. The method of claim 35, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

37. The method of any one of claims 35 to 36, wherein desmopressin is first administered to the subject followed by temozolomide administration to the subject.

38. The method of any one of claims 35 to 37, wherein temozolomide is administered from about 3 days to about 7 days following desmopressin administration, from about 2 days to about 10 days following desmopressin administration, from about 1 days to about 12 days following desmopressin administration, from about 30 minutes to about 60 minutes following desmopressin administration.

39. The method of any one of claims 29 to 38, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

40. The method of claim 29, further comprising:- administering desmopressin prior to surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery; and- administering desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma.

41. The method of claim 40, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg.

42. The method of claim 40 or 41 , further comprising administering temozolomide to said subject.

43. The method of claim 42, where said temozolomide is administered at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy.

44. The method of any one of claim 29 to 43, wherein said subject is a human.

45. A method for treating or preventing glioblastoma in a subject, comprising, or consisting of, measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject;and administering to a subject in need thereof a therapeutically effective amount of a vasopressin analogue, optionally in a physiological carrier or a pharmaceutically acceptable salt thereof, when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

46. The method of claim 45, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

47. The method of claim 45 or 46, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

48. The method of any one of claims 45 to 47, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

49. The method of any one of claims 45 to 48, wherein vasopressin analog is administered before, during, and / or after surgery to remove the glioblastoma.

50. The method of any one of claims 45 to 49, wherein vasopressin analog is administered perioperatively.

51. The method of any one of claims 45 to 50, wherein the vasopressin analog is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

52. The method of any one of claims 45 to 51 , further comprising administering a therapeutically effective amount of temozolomide.

53. The method of claim 52, wherein temozolomide administered at a dose of from 75 to 200 mg / m2daily.

54. The method of any one of claims 45 to 53, wherein vasopressin analog is first administered to the subject followed by temozolomide administration.

55. The method of any one of claims 45 to 54, wherein temozolomide is administered from about 3 days to about 7 days following desmopressin administration, from about 2 days to about 10 days following desmopressin administration, from about 1 days to about 12 days following desmopressin administration, from about 30 minutes to about 60 minutes following desmopressin administration.

56. The method of any one of claims 45 to 55, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

57. The method of any one of claims 45 - 56, wherein the subject is a human.

58. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising ,or consisting of:- administering desmopressin prior to the surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery;- administering desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma; and- administration to said subject temozolomide at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy;- administering temozolomide for 6 cycles of maintenance treatment; and- administering desmopressin on day 1 of each said cycle.

59. The method of claim 58, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

60. The method of claim 58 or 59, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

61. The method of any one of claims 58 to 60, wherein desmopressin is administered at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

62. The method of any one of claims 58 to 61 , wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

63. The method of any one of claims 58 to 62, wherein said subject is a human.

64. A kit for treating glioblastoma in a human, comprising or consisting of: desmopressin and TMZ, and optionally a container and optionally instructions for the use thereof.

65. Use of a therapeutically effective amount of desmopressin for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma.

66. The use of claim 65, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

67. The use of claim 65 or 66, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

68. The use of any one of claims 65 to 67, wherein desmopressin is for administration before, during, and / or after surgery to remove the glioblastoma.

69. The use of any one of claims 65 to 68, wherein desmopressin is for administration perioperatively.

70. The use of any one of claims 65 to 69, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

71. The use of any one of claims 65 to 70, further comprising use of a therapeutically effective amount of temozolomide.

72. The use of claim 71, wherein temozolomide is for administration at a dose of from 75 to 200 mg / m2daily.

73. The use of any one of claims 71 to 72, wherein desmopressin is for administration first followed by temozolomide administration.

74. The use of claim 73, wherein temozolomide is for administration from about 3 days to about 7 days following use of desmopressin, from about 2 days to about 10 days following use of desmopressin, from about 1 day to about 12 days following use of desmopressin, from about 30 minutes to about 60 minutes following use of desmopressin.

75. The use of any one of claims 65 - 74, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

76. Use of desmopressin prior to surgery to remove some or all of a glioblastoma, preferably about 30 minutes to about 60 before surgery; and- administering use of desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma, for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma.77 The use of claim 76, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg78. The use of claim 76 or 77, further comprising use of temozolomide to said subject.

79. The use of claim 78, where said temozolomide is for administration at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy.

80. A method of treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, comprising or consisting of: administering Use of a therapeutically effective amount of a vasopressin analogue for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma.81 The use of any one of claim 80, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

82. The use of claim 80 or 81 wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

83. The use of any one of claim 80 to 82, wherein vasopressin analogue is for administration before, during, and / or after surgery to remove the glioblastoma.

84. The use of any one of claims 80 to 83, wherein the vasopressin analogue is for administration perioperatively.

85. The use of any one of claims 80 to 84, wherein the vasopressin analogue is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

86. The use of any one of claims 80 to 85, further comprising use of a therapeutically effective amount of temozolomide.

87. The use of claim 86, wherein temozolomide is for administration at a dose of from 75 to 200 mg / m2daily.

88. The use of any one of claims 86 to 87, wherein the vasopressin analogue is for administration first followed by use of temozolomide.

89. The use of any one of claims 86 to 88, wherein temozolomide is for administration from about 3 days to about 7 days following use of the vasopressin analogue, from about 2 days to about 10 days following use of the vasopressin analogue, from about 1 days to about 12 days following use of the vasopressin analogue, from about 30 minutes to about 60 minutes following use of the vasopressin analogue.

90. The use of any one of claims 80 - 89, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

91. The use of any one of claims 80 to 90, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

92. The use of any one of claims 65 to 91 , wherein the subject is a human.

93. Use of a therapeutically effective amount of desmopressin for treating or preventing glioblastoma in a subject, or in the manufacture of a medicament for treating or preventing glioblastoma in a subject, comprising measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, wherein the use of desmopressin is indicated when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

94. The use of claim 93, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, ora glioblastoma multiforme.

95. The use of claim 94, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

96. The use of any one of claims 93 or 95, wherein desmopressin is for administration before, during, and / or after surgery to remove the glioblastoma.

97. The use of any one of claims 93 to 96, wherein desmopressin is for administration perioperatively.

98. The use of any one of claims 93 to 97, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

99. The use of any one of claims 93 to 98, further comprising use of a therapeutically effective amount of temozolomide.

100. The use of claim 99, wherein temozolomide is administered at a dose of from 75 to 200 mg / m2daily.

101. The use of any one of claims 99 to 100, wherein desmopressin is for administration first followed by use of temozolomide.

102. The use of any one of claims 99 to 101, wherein temozolomide is for administration from about 3 days to about 7 days following use of desmopressin, from about 2 days to about 10 days following use of desmopressin, from about 1 days to about 12 days following use of desmopressin, from about 30 minutes to about 60 minutes following use of desmopressin.

103. The use of any one of claims 93 to 102, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

104. The use of claim 93, further comprising:- use of desmopressin prior to surgery to remove some or all of the glioblastoma, preferably about 30 minutes to about 60 before surgery; and- use of desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma.

105. The use of claim 104, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, or about 0.3 pg / kg, or about 0.4 pg / kg.

106. The use of claim 104 or 105, further comprising use of temozolomide.

107. The use of claim 106, where said temozolomide is for administration at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy.

108. The use of any one of claim 93 to 107, wherein said subject is a human.

109. Use of a therapeutically effective amount of of a vasopressin analogue for treating or preventing glioblastoma in a subject, or in the manufacture of a medicament for treating or preventing glioblastoma in a subject, comprising measuring an amount of arginine vasopressin type 2 receptor (AVPR2) polypeptide, AVPR2 activity, or AVPR2 mRNA, in a sample from a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, wherein the use of ther vasopressin analogue is indicated when the amount of AVPR2 polypeptide, or AVPR2 activity, or AVPR2 mRNA is high, optionally compared to a control.

110. The use of claim 109, wherein the vasopressin analogue is desmopressin, synthetic arginine vasopressin, lysine vasopressin, terlipressin, felypressin, or ornipressin.

111. The use of claim 109 or 110, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

112. The use of any one of claims 109 to 111, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or +7 / -10 chromosomal signature.

113. The use of any one of claims 109 to 112, wherein vasopressin analog is for administration before, during, and / or after surgery to remove the glioblastoma.

114. The use of any one of claims 109 to 113, wherein vasopressin analog is for administration perioperatively.

115. The use of any one of claims 109 to 114, wherein the vasopressin analog is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

116. The use of any one of claims 109 to 117, further comprising use of a therapeutically effective amount of temozolomide.

117. The use of claim 116, wherein temozolomide for administration at a dose of from 75 to 200 mg / m2daily.

118. The use of any one of claims 109 to 117, wherein vasopressin analog is for administration first followed by use of temozolomide administration.

119. The use of any one of claims 109 to 118, wherein temozolomide is for administration from about 3 days to about 7 days following use of desmopressin, from about 2 days to about 10 days following use of desmopressin, from about 1 days to about 12 days following use of desmopressin, from about 30 minutes to about 60 minutes following use of desmopressin.

120. The use of any one of claims 109 to 119, wherein the glioblastoma is primary glioblastoma or recurrent glioblastoma.

121. The use of any one of claims 109 - 120, wherein the subject is a human.

122. Use of desmopressin prior to surgery to remove some or all of a glioblastoma in a subject, preferably about 30 minutes to about 60 before surgery for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma, or in the manufacture of a medicament for treating a subject with glioblastoma, at risk of developing glioblastoma, or suspected of having glioblastoma;- use of desmopressin about 24 hour following said surgery to remove some or all of the glioblastoma;- use of temozolomide at a dose of about 75 mg / m2daily for about 42 days concomitant with radiotherapy;- use of temozolomide for 6 cycles of maintenance treatment; and- use of desmopressin on day 1 of each said cycle.

123. The use of claim 122, wherein the glioblastoma is a giant cell glioblastoma, a small cell glioblastoma, or a glioblastoma multiforme.

124. The use of claim 122 or 123, wherein the glioblastoma is an IDH-wildtype diffuse astrocytic tumour with at least one of the following molecular markers: TERT promoter mutations, EGFR amplification, or+7 / -10 chromosomal signature.

125. The use of any one of claims 122 to 124, wherein desmopressin is for administration at a dose of between about 0.1 pg / kg to about 1.0 pg / kg, about 0.2 pg / kg to about 0.5 pg / kg, about 0.3 to about 0.4 pg / kg, about 0.3 pg / kg, or about 0.4 pg / kg.

126. The use of any one of claims 122 to 125, wherein temozolomide is for administration at a dose of from 75 to 200 mg / m2daily.

127. The use of any one of claims 122 to 126, wherein said subject is a human.

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