Methods of modulating erythropoiesis using beta-2 adrenergic receptor antagonists

Abstract

The present disclosure provides, among other things, methods, compositions, and uses for treating diseases or disorders characterized by elevated red blood cells using β2 adrenergic receptor antagonists such as β2-specific adrenergic receptor antagonists (e.g., ICI-118551, butaxamine, or pharmaceutically acceptable salts thereof) or β1 / β2-specific adrenergic receptor antagonists (e.g., propranolol or a pharmaceutically acceptable salt thereof). Also provided are methods, compositions, and uses for decreasing erythroid differentiation using the same. Provided methods of treatment comprise administering an effective amount of a β2-specific adrenergic receptor antagonist or a β1 / β2-specific adrenergic receptor antagonist to a patient in need thereof. Also provided are methods, compositions, and uses for ICI-118551 in treating splenomegaly, splenic erythroid hyperplasia, and / or splenic architecture effacement.

Description

METHODS OF MODULATING ERYTHROPOIESIS USING BETA-2 ADRENERGIC RECEPTOR ANTAGONISTSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 641,922, filed May 2, 2024, and U.S. Provisional Application No. 63 / 720,609, filed November 14, 2024, the disclosures of each of which are incorporated by reference herein in their entirety.BACKGROUND

[0002] Myeloproliferative disorders (MPDs), myeloproliferative neoplasms (MPNs), and secondary erythrocytosis disorders are conditions generally characterized by chronic increases in some or all of the blood cells (e.g., platelets, white blood cells, and red blood cells). This group of blood disorders includes, e.g., polycythemia vera (PV), essential thrombocythemia (ET), myelofibrosis (e.g., primary myelofibrosis, post-polycythemia vera myelofibrosis, and post-essential thrombocythemia myelofibrosis), chronic myeloid leukemia (CML), and secondary polycythemia. The existing lines of treatment for MPDs including PV are very limited. Patients with PV remain predominantly dependent on frequent blood withdrawals, or hydroxyurea / Jak-inhibitor treatment regimens which produce undue toxicities in patients. Thus, there is an unmet need to identify additional therapies to treat disorders associated with elevated red blood cells and complications arising from these disorders.SUMMARY

[0003] The present invention(s) described herein is based, in part, on the discovery of a therapeutic approach for disorders or other diseases associated with elevated erythropoiesis by administering a P2 adrenergic receptor antagonist, such as, e.g., ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing, e.g., including, but not limited to, ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride.

[0004] In some embodiments, provided are methods for treating a disease or disorder associated with elevated red blood cells using a p2-specific adrenergic receptor antagonist. In some embodiments, a p2-specific adrenergic receptor antagonist is selected from ICI-118551, butaxamine, and pharmaceutically acceptable salts thereof.

[0005] ICI-118551 is a selective antagonist of beta-2 adrenergic receptor (ADRB2) that is employed as an ocular hypotensive agent in ophthalmic disease. Butaxamine, also known as butoxamine, is also a selective antagonist of ADRB2 that has been primarily used in experimental situations.

[0006] In some embodiments, provided are methods for treating a disease or disorder associated with elevated red blood cells using a pi / p2-specific adrenergic receptor antagonist. In some embodiments, a pi / p2-specific adrenergic receptor antagonist is selected from propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, a pi / p2-specific adrenergic receptor antagonist is propranolol or a pharmaceutically acceptable salt thereof. Propranolol is a non- selective P-adrenergic receptor antagonist that is commonly used to treat a number of conditions including, e.g., hypertension, irregular heart rate, and essential tremors.

[0007] The present disclosure encompasses a new and unexpected finding that P2- specific and pi / p2-specific adrenergic receptor antagonists (e.g., ICI-118551, butaxamine, propranolol, and pharmaceutically acceptable salts thereof) can decrease erythroid (red blood cell) parameters in vivo without leading to anemia. The disclosed approach has the potential benefit of normalizing excess red blood cells without producing toxic side effects.

[0008] The present disclosure also provides the insight that ICI-118551 can be used to treat splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement associated with red blood cell disorders. In some embodiments, ICI-118551 can be used to normalize splenic size and splenic architecture in PV patients.

[0009] In some embodiments, provided are methods of treating a disease or disorder associated with elevated red blood cells in a patient in need thereof, the method comprising administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing.

[0010] In some embodiments, provided are methods of decreasing red blood cells in a patient, the method comprising administering to the patient an effective amount of a P 1 / |32- specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist. In some embodiments, provided are methods of decreasing red blood cells in a patient, the method comprising administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing.

[0011] In some embodiments, provided are methods of decreasing erythroid differentiation in a patient, the method comprising administering to the patient an effectiveamount of a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist. In some embodiments, provided are methods of decreasing erythroid differentiation in a patient, the method comprising administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing.

[0012] In some embodiments, provided are methods of treating elevated red blood cells in a patient, the method comprising administering to the patient an effective amount of a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist. In some embodiments, provided are methods of treating elevated red blood cells in a patient, the method comprising administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing.

[0013] In some embodiments, provided are methods of treating splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement in a patient having elevated red blood cells, the method comprising administering to the patient an effective amount of ICI- 118551 or a pharmaceutically acceptable salt thereof.

[0014] In some embodiments, a patient has a disease or disorder associated with elevated red blood cells that is a myeloproliferative disorder. In some embodiments, a myeloproliferative disorder is polycythemia vera (PV). In some embodiments, a myeloproliferative disorder has progressed to a cancer.

[0015] In some embodiments, a patient has a myeloproliferative disorder associated with & .JAK2 mutation. In some embodiments, the JAK2 mutation is &.Jctk2V6I7k' mutation associated with elevated red blood cell production, PV and / or polycythemia.

[0016] In some embodiments, a patient has PV associated with a JAK2 mutation. In some embodiments, a patient has PV associated with a Jak2V617F mutation.

[0017] In some embodiments, provided are methods that treat both PV and hypertension by administering propranolol or a pharmaceutically acceptable salt thereof, e.g. propranolol hydrochloride.

[0018] In some embodiments, a patient has secondary polycythemia. In some embodiments, a patient has secondary polycythemia caused by sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastomaand meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

[0019] In some embodiments, provided are methods that treat both secondary polycythemia and hypertension by administering an effective amount of a pi / p2-specific adrenergic receptor antagonist. In some embodiments, provided are methods that treat both secondary polycythemia and hypertension by administering propranolol or a pharmaceutically acceptable salt thereof, e.g. including, but not limited to, propranolol hydrochloride.

[0020] In some embodiments, a patient has splenomegaly. In some embodiments, a patient has splenic erythroid hyperplasia. In some embodiments, a patient has splenic architecture effacement. In some embodiments, administering to the patient an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof treats the splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement.

[0021] In some embodiments, administering to the patient an effective amount of a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist decreases red blood cells in the patient. In some embodiments, administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases red blood cells in the patient.

[0022] In some embodiments, a patient has an elevated red blood cell mass. In some embodiments, an elevated red blood cell mass is greater than 29 mL / kg for a female patient and / or greater than 32 mL / kg for a male patient. In some embodiments, administering to the patient an effective amount of a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist decreases the red blood cell mass in a patient. In some embodiments, administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases the red blood cell mass in a patient. In some embodiments, administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases the red blood cell mass to less than 29 mL / kg for a female patient and / or less than 32 mL / kg for a male patient.

[0023] In some embodiments, a patient has elevated hematocrit. In some embodiments, elevated hematocrit is greater than 48% in a female patient and / or greater than 51% in a male patient. In some embodiments, administering to the patient an effective amount of a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptorantagonist decreases the hematocrit value in a patient. In some embodiments, administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases the hematocrit value in a patient. In some embodiments, administering to the patient an effective amount of ICI- 118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases the hematocrit value to less than 48% for a female patient and / or less than 51% for a male patient.

[0024] In some embodiments, a patient has elevated hemoglobin. In some embodiments, elevated hemoglobin is greater than 165 g / L in a female patient and / or greater than 185g / L in a male patient. In some embodiments, administering to the patient an effective amount of a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist decreases the hemoglobin value in a patient. In some embodiments, administering to the patient an effective amount of ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases the hemoglobin value in a patient. In some embodiments, administering to the patient an effective amount of ICI- 118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing decreases the hemoglobin value to less than 165 g / L for a female patient and / or less than 185g / L for a male patient.

[0025] In some embodiments, the method does not induce anemia in the patient.

[0026] In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof is administered to the patient. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof is orally administered to the patient. In some embodiments, a pharmaceutically acceptable salt of ICI-118551 is ICI-118551 hydrochloride.

[0027] In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof is administered to the patient. In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof is orally administered to the patient. In some embodiments, a pharmaceutically acceptable salt of butaxamine is butaxamine hydrochloride.

[0028] In some embodiments, propranolol or a pharmaceutically acceptable salt thereof is administered to the patient. In some embodiments, propranolol or a pharmaceutically acceptable salt thereof is orally administered to the patient. In some embodiments, a pharmaceutically acceptable salt of propranolol is propranolol hydrochloride.

[0029] In some embodiments, the patient is a human.

[0030] In some embodiments, the patient has received previous treatment for the disease or disorder associated with elevated red blood cells. In some embodiments, the patient has not received previous treatment for the disease or disorder associated with elevated red blood cells. In some embodiments, a previous treatment for the disease or disorder associated with elevated red blood cells is selected from chemotherapy (e.g., hydroxyurea), blood draws, or treatment with a JAK2 inhibitor.

[0031] In some embodiments, provided are pharmaceutical compositions comprising a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist for use in a method as described herein. In some embodiments, provided are pharmaceutical compositions comprising ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing for use in a method as described herein.

[0032] In some embodiments, provided are uses of pi / p2-specific adrenergic receptor antagonists or p2-specific adrenergic receptor antagonists in the manufacture of a medicament for treating a disease or disorder associated with elevated red blood cells.

[0033] In some embodiments, provided are uses of ICI-118551 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease or disorder associated with elevated red blood cells. In some embodiments, the medicament further treats splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement.

[0034] In some embodiments, provided are uses of butaxamine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a disease or disorder associated with elevated red blood cells. In some embodiments, provided are uses of propranolol or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease or disorder associated with elevated red blood cells.

[0035] Throughout the description, where methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are methods according to the present disclosure that consist essentially of, or consist of, the recited steps.

[0036] It should be understood that the order of steps or order for performing certain action is immaterial so long as the invention remains operable. Moreover, two or more steps or actions may be conducted simultaneously.

[0037] Any of the embodiments described herein can be used in conjunction with one another, unless otherwise indicated or apparent from the context. Other embodiments will become apparent to those skilled in the art from a review of the ensuing description.BRIEF DESCRIPTION OF THE DRAWINGS

[0038] The Drawings included herein, which are composed of the following Figures, are for illustration purposes only and not for limitation.

[0039] FIG. 1A shows that ICI-118551 and propranolol dampen erythropoiesis. Administration of ICI-118551 (ICI) and propranolol (Pro) at 5 pM to primary human HSPCs impair erythroid differentiation (CD235a). ICI-118551 dampens erythropoiesis more potently than Propranolol. Representative of n=5 healthy donor-derived HSPCs; n=4 technical replicates within each donor. **** p < 0.0001, one-way analysis of variance (ANOVA). Data represented as mean ± SEM. All comparisons are done with respect to control (DMSO treated). ns= non-significant.

[0040] FIG. IB shows that antagonists of beta-2 adrenergic receptors do not affect megakaryopoiesis. Administration of ICI-118551 (ICI) or propranolol (Pro) at 1 / 5 pM to primary human HSPCs does not influence megakaryocytic differentiation (CD41 / 61 staining). Representative of n=5 healthy donor-derived HSPCs; n=4 technical replicates within each donor. One-way analysis of variance (ANOVA). Data represented as mean ± SEM. All comparisons are done with respect to control (DMSO treated). ns= non-significant.

[0041] FIG. 2 provides a schematic of ICI treatment via intraperitoneal (i.p.) administration in wild-type mice at 0.5 mg / kg daily for 30 days.

[0042] FIG. 3A-FIG. 3C show that ICI treatment via i.p. administration for 30 days decreases RBC parameters in wild-type mice at steady state. ICI treatment decreases RBC parameters: FIG. 3 A shows a decrease in RBCs, FIG. 3B shows a decrease in hematocrit (HCT)%, and FIG. 3C shows a decrease in hemoglobin (Hb) in the peripheral blood of wildtype mice after 30 days of daily i.p. administrations. * p < 0.05, *** p < 0.001, **** p < 0.0001, ANOVA. n=5 male mice per group. All comparisons are done with respect to vehicle (DMSO) treated controls.

[0043] FIG. 4A-FIG. 4D show that ICI treatment daily via i.p. administration for 30 days does not affect WBC parameters or platelets in wild-type mice at steady state. ICI treatment does not affect WBCs (FIG. 4A), platelets (FIG. 4B), monocytes (FIG. 4C), and lymphocytes (FIG. 4D) in the peripheral blood after 30 days of daily i.p. administration. ANOVA. n=5 male mice per group. All comparisons are done with respect to vehicle (DMSO) treated controls, ns = non-significant.

[0044] FIG. 5A and FIG. 5B show that ICI treatment daily via i.p. administration for 30 days decreases RBC parameters in wild-type mice at steady state but does not lead toanemia. ICI treatment mildly decreases hemoglobin (Hb) (FIG. 5 A) and hematocrit (HCT)% (FIG. 5B) in the peripheral blood after 30 days of daily i.p. administration, but does not lead to anemia. * p < 0.05, ANOVA. n=5 male mice per group. All comparisons are done with respect to vehicle (DMSO) treated controls.

[0045] FIG. 6A and FIG. 6B show that ICI treatment 3x weekly i.p. administration normalizes RBC parameters in Erythropoietin (EPO) treated mice. ICI treatment 0.5 mg / kg decreases (FIG. 6A) Hemoglobin (Hb) and (FIG. 6B) Hematocrit (HCT) in the peripheral blood of mice treated with EPO (EPO treatment: 500 lU / kg 3x weekly i.p. dose). * p < 0.05, ** p < 0.01, *** p < 0.001, ANOVA. n=5 female mice per group. Vehicle (Veh): DMSO treated controls.

[0046] FIG. 7 shows ICI treatment 5x weekly for 6 weeks i.p. normalizes RBC parameters in Polycythemia Vera (PV) model mice. ICI treatment 0.5 mg / kg i.p. 5x weekly normalizes Hemoglobin (Hb) in the peripheral blood of PV model mice carrying Jak2V617F mutation in their hematopoietic compartment. Vehicle: DMSO (dm). LC: littermate controls.

[0047] FIG. 8A-FIG. 8C show ICI treatment 5x weekly for 6 weeks i.p. decreases erythroid progenitors in the BM and spleen of PV model mice. ICI treatment 0.5 mg / kg 5x weekly i.p. administration reduces erythroid progenitors in the BM (FIG. 8A), colonyforming unit-erythroid (CFU-e) in the BM (FIG. 8B), and erythroid progenitors in the spleen (FIG. 8C) of PV model mice. Vehicle : DMSO (dm). LC: littermate controls. * p < 0.05, ** p < 0.01, *** P < 0.001, **** p < 0.0001, ANOVA. All comparisons are done with respect to PV mice administered vehicle (DMSO: dm) treated controls.

[0048] FIG. 9 A and FIG. 9B show ICI treatment 5x weekly for 6 weeks i.p. administration reduces splenomegaly in PV model mice. ICI treatment 0.5 mg / kg 5x weekly i.p. administration reduces spleen size (FIG. 9A) and spleen weight (FIG. 9B) in PV model mice. Vehicle : DMSO (dm). LC: littermate controls. * p < 0.05, ** p < 0.01, *** P < 0.001, **** p < 0.0001, ANOVA. All comparisons are done with respect to PV mice administered vehicle (DMSO: dm) treated controls.

[0049] FIG. 10 shows ICI treatment 5x weekly for 6 weeks i.p. administration normalizes splenic architecture in PV model mice. Histopathologic H&E stains showing erythroid hyperplasia and complete effacement of splenic architecture in PV mice, and improvement towards normalcy in ICI-treated PV mice. Vehicle: DMSO (dm). LC: littermate controls.

[0050] FIG. 11A-FIG.11C show the effects of treatment with a selective P2 antagonist, Butaxamine, on primary human HSPCs on erythroid differentiation in vitro. FIG. 11 A shows that administration of butaxamine (Buta) at 2 / 5 pM to primary human HSPCs potently dampens erythroid differentiation (CD235a). FIG. 1 IB and FIG. 11C show that Buta does not impact myelopoiesis (CD1 lb staining) or megakaryopoiesis (CD41 / 61 staining), respectively. n=4 technical replicates within each donor. ** p < 0.01, one-way analysis of variance (ANOVA). Data represented as mean ± SEM. All comparisons are done with respect to control (DMSO) treatment, ns = non-significant.

[0051] FIG. 12 shows an experimental plan to analyze the effect of butaxamine in an EPO-induced model of secondary polycythemia. Model of secondary polycythemia created by EPO 500 lU / kg treatment 3x weekly i.p. administration; butaxamine was administered at 0.5 mg / kg 3 times per week (3xw) i.p. administration (0.5 mg / kg 3xw i.p.).

[0052] FIG. 13A and FIG. 13B show that butaxamine (Buta) treatment normalizes Erythropoietin (EPO)-induced RBC parameters in vivo. Buta treatment at 0.5 mg / kg 3x weekly i.p. normalizes RBC parameters in Erythropoietin treated mice (EPO: 500IU / kg 3x weekly i.p. dose). Buta treatment decreases (FIG. 13A) RBCs and (FIG. 13B) Hemoglobin (Hb) in the peripheral blood of mice treated with Erythropoietin (EPO). * p < 0.05, ** p < 0.01, *** p < 0.001, ANOVA. n=5 female mice per group. Vehicle (Veh): DMSO. All comparisons are shown with respect to the “Epo only” group.

[0053] FIG. 14 shows that butaxamine treatment does not influence body weights in vivo. Specifically, butaxamine treatment (0.5 mg / kg 3xw i.p.) of mice did not significantly increase or decrease body weight over a period of up to 40 days treatment.

[0054] FIG. 15 shows an experimental plan to analyze the effect of butaxamine (Buta)- 5xw i.p. in Polycythemia Vera mouse model. Model of Polycythemia Vera: Jak2V617F mutant mice; Butaxamine treatment: 0.5 mg / kg 5xw i.p.

[0055] FIG. 16 shows that butaxamine treatment normalizes hemoglobin in peripheral blood of Polycythemia Vera (PV)-mouse model (Jak2V617F mutant mice). Buta treatment 0.5 mg / kg i.p. 5x weekly normalizes Hemoglobin (Hb) in the peripheral blood of Polycythemia Vera mice carrying Jak2V617F mutation in their hematopoietic compartment. Vehicle : DMSO (dm). LC: littermate controls. LC+Buta0.5: n=10; LC+Veh: n=10; PV+Veh: n=3; PV+Buta0.5: n=5.

[0056] FIG. 17A-FIG.17B show that butaxamine treatment normalizes RBCs (FIG. 17A) and hematocrit (HCT, FIG. 17B) in peripheral blood of Polycythemia Vera (PV) micecarrying Jak2V617F mutation in their hematopoietic compartment. Vehicle : DMSO (dm). LC: littermate controls. LC+Buta0.5: n=10; LC+Veh: n=10; PV+Veh: n=3; PV+Buta0.5: n=5.

[0057] FIG. 18A-FIG. 18B show that butaxamine dampens erythroid progenitors in the bone marrow (BM) of PV-mouse model (Jak2V617F mutant mice). Buta treatment 0.5 mg / kg 5x weekly i.p. for 4 weeks reduces (FIG. 18A) erythroid progenitors in the BM, (FIG. 18B) colony-forming unit-erythroid (CFU-e) in the BM of PV mice. Vehicle : DMSO (dm). LC: littermate controls. * p < 0.05, **** p < 0.0001, ANOVA. LC+Buta0.5: n=10; LC+Veh: n=10; PV+Veh: n=3; PV+Buta0.5: n=5.

[0058] FIG. 19 shows that butaxamine dampens erythroid progenitors in the spleen of PV-mouse model (Jak2V617F mutant mice). Buta treatment 0.5 mg / kg 5x weekly i.p. for 4 weeks reduces erythroid progenitors in the spleen of PV mice. Vehicle : DMSO (dm). LC: littermate controls. * p < 0.05, **** p < 0.0001, ANOVA. LC+Buta0.5: n=10; LC+Veh: n=10; PV+Veh: n=3; PV+Buta0.5: n=5.

[0059] FIG. 20A-FIG. 20B show that butaxamine does not impact myelopoiesis or megakaryopoiesis in the BM of PV-mouse model (Jak2V617F mutant mice). Buta treatment 0.5 mg / kg 5x weekly for 4 weeks i.p. does not impact (FIG. 20A) granulocytic monocyte progenitors (GMP) or (FIG. 20B) megakaryocytic progenitors (MkP) in the BM of PV mice. Vehicle : DMSO (dm). LC: littermate controls, ns: non-significant, ANOVA. LC+Buta0.5: n=10; LC+Veh: n=10; PV+Veh: n=3; PV+Buta0.5: n=5.

[0060] FIG. 21 shows that propranolol treatment normalizes hemoglobin (Hb) in peripheral blood of Polycythemia Vera (PV)-mouse model (Jak2V617F mutant mice). Propranolol treatment 0.3 mg / kg i.p. 5x weekly normalizes Hb in the peripheral blood of PV mice carrying Jak2 V617F mutation in their hematopoietic compartment. Vehicle : DMSO (dm). LC: littermate controls. Control+Veh: n=5; PV+Veh: n=3; PV+Pro0.3: n=3.

[0061] FIG. 22 shows an experimental plan to analyze the effect of ICL118551 (ICI) vs. butaxamine (Buta) vs. propranolol (Pro) (0.5 mg / kg i.p. 3x weekly) in EPO (500IU / kg 3xweekly ip)-induced mouse model of secondary Polycythemia).

[0062] FIG. 23A-FIG. 23B show that ICI 118551 (ICI), Butaxamine (Buta), Propranolol (Pro) treatments 3x weekly i.p. normalize RBC parameters in Erythropoietin- induced mouse model of secondary Polycythemia. ICI, Buta, and Pro treatments 0.5 mg / kg 3x weekly i.p. decreases RBCs (FIG. 23 A) and Hematocrit (HCT; FIG. 23B) in the peripheral blood of mice treated with Erythropoietin (EPO: 500IU / kg 3x weekly i.p. dose). ICI , Buta,and Pro showed similar potency in their effects on both RBCs and HCT. * p < 0.05, ** p < 0.01, ANOVA. n=5 female mice per group, ns: non-significant. Vehicle (Veh): DMSO.

[0063] FIG. 24A-FIG. 24B show that ICI 118551 (ICI), Butaxamine (Buta), Propranolol (Pro) treatments 3x weekly i.p. normalize RBC parameters in Erythropoietin (EPO: 500IU / kg 3x weekly i.p. dose)- induced mouse model of secondary Polycythemia. ICI, Buta, and Pro treatments 0.5 mg / kg decreases hemoglobin (Hb), both % change in Hb (FIG. 24A) and absolute Hb (FIG. 24B), in the peripheral blood of mice treated with Erythropoietin (EPO). *** p < 0.001, **** p < 0.0001, ANOVA. n=5 female mice per group, ns: nonsignificant. Vehicle (Veh): DMSO.

[0064] FIG. 25A-FIG. 25B show that ICI 118551 (ICI), Butaxamine (Buta), or Propranolol (Pro) treatments 3x weekly i.p. do not affect WBCs or platelets in Erythropoietin (EPO: 500IU / kg 3x weekly i.p. dose)- induced mouse model of secondary Polycythemia. ICI, Buta, and Pro treatments 0.5 mg / kg 3x weekly i.p. do not affect WBCs (FIG. 25A) or platelets (FIG. 25B) in the peripheral blood of mice treated with Erythropoietin (EPO). ANOVA. n=5 female mice per group. Vehicle (Veh): DMSO.DETAILED DESCRIPTIONDefinitions

[0065] In order for the present invention(s) to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. The publications and other reference materials referenced herein to describe the background of the disclosure and to provide additional detail regarding its practice are hereby incorporated by reference.

[0066] In this application, unless otherwise clear from context, (i) the terms “a” and “an” are used herein to refer to one or to more than one (z.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0067] The term “administering" means providing a pharmaceutical agent or composition to a subject, and includes, but is not limited to, administration by a medical professional and self-administration. This involves the physical introduction of a composition comprising a therapeutic agent to a subject, using any of the various methods and delivery systems known to those skilled in the art. Administering may be performed, for example, once, a plurality of times, and / or over one or more extended periods.

[0068] The terms “cancer” or “tumor” or “hyperproliferative” refer to the presence of cells possessing characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features.

[0069] Cancer cells are often in the form of a tumor, but such cells may exist alone within an animal, or may be a non-tumorigenic cancer cell, such as a leukemia cell. As used herein, the term “cancer” includes premalignant as well as malignant cancers. Cancers include, but are not limited to, B cell cancer, e.g., myelomas like multiple myeloma, Waldenstrom's macroglobulinemia, the heavy chain diseases, such as, for example, alpha chain disease, gamma chain disease, and mu chain disease, benign monoclonal gammopathy, and immunocytic amyloidosis, melanomas, breast cancer, lung cancer, bronchus cancer, colorectal cancer, prostate cancer, pancreatic cancer, stomach cancer, ovarian cancer, urinary bladder cancer, brain or central nervous system cancer, peripheral nervous system cancer, esophageal cancer, cervical cancer, uterine or endometrial cancer, cancer of the oral cavity or pharynx, liver cancer, kidney cancer, testicular cancer, biliary tract cancer, small bowel or appendix cancer, salivary gland cancer, thyroid gland cancer, adrenal gland cancer, osteosarcoma, chondrosarcoma, cancer of hematologic tissues, and the like. Other nonlimiting examples of types of cancers applicable to the methods encompassed by the present disclosure include human sarcomas and carcinomas, e.g., fibrosarcoma, myosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, liver cancer, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, bone cancer, brain tumor, testicular cancer, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma; leukemias, e.g., acute lymphocytic leukemia and acute myelocytic leukemia (myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia); chronic leukemia (chronic myelocytic(granulocytic) leukemia and chronic lymphocytic leukemia); and polycythemia vera, lymphoma (Hodgkin's disease and non-Hodgkin's disease), myeloma, multiple myeloma, Waldenstrom's macroglobulinemia, and heavy chain disease. In some embodiments, cancers are epithelial in nature and include but are not limited to, bladder cancer, breast cancer, cervical cancer, colon cancer, gynecologic cancers, renal cancer, laryngeal cancer, lung cancer, oral cancer, head and neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, or skin cancer. In other embodiments, the cancer is breast cancer, prostate cancer, lung cancer, or colon cancer. In still other embodiments, the epithelial cancer is non-small-cell lung cancer, nonpapillary renal cell carcinoma, cervical carcinoma, ovarian carcinoma (e.g., serous ovarian carcinoma), or breast carcinoma. The epithelial cancers may be characterized in various other ways including, but not limited to, serous, endometrioid, mucinous, clear cell, Brenner, or undifferentiated.

[0070] The term “erythroid progenitor cell” refers to the hematopoietic stem cell- derived progenitor cell that gives rise to erythrocytes (red blood cells) after terminal differentiation.

[0071] The term “patient”, as used herein, refers to a human suffering from disease or disorder characterized by elevated red blood cells.

[0072] The term “therapeutic effect” refers to a local or systemic effect in humans, caused by a pharmacologically active substance. The term thus means any substance intended for use in the diagnosis, cure, mitigation, or treatment of disease or in the enhancement of desirable physical or mental development and conditions in a patient (e.g., a human suffering from a disease or disorder associated with elevated red blood cells).

[0073] The terms “therapeutically-effective amount” and “effective amount” refers to an amount necessary (for example, at dosages and for periods of time and for the means of enteral or oral administration) to achieve the desired therapeutic result of treating a disease or disorder associated with elevated red blood cells. An effective amount of the antagonist of P2-adrenoreceptor (e.g., ICI-118551, butaxamine, propranolol) may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the monoclonal antibody to elicit a desired response in the individual. An effective amount is also one in which medical provider, e.g., the attending physician, determines that any toxic or detrimental effects of the antagonist are outweighed by the therapeutically beneficial effects.

[0074] As used hererin, “treating” a disease, disorder or condition in a subject or “treating” a subject having a disease, disorder or condition refers to subjecting the subject to apharmaceutical treatment, e.g., administration of a composition, such that at least one symptom of the disease, disorder, or condition is decreased or prevented from worsening.P Adrenergic Receptor Antagonists

[0075] The present disclosure describes, inter alia, compositions, methods and uses for treating disorders or other diseases associated with elevated red blood cells by administering a P2 adrenergic receptor antagonist. Beta-adrenergic receptors are transmembrane glycoprotein structures that elicit a response inside the cell when interacting with catecholamines. They belong to a major receptor family (R7G) containing other receptors responding to substances other than catecholamines, and these receptors are coupled with guanine nucleotide (GTP) binding proteins (G proteins). Beta receptors divide into three subtypes; beta-1 (P 1 ), beta-2 (P2), and beta-3 (P3). Other adrenergic receptors are alpha-1 (al) and alpha-2 (a2) receptors. Abosamak and Shahin, Beta2 Receptor Agonists and Antagonists. [Updated 2023 Jul 3], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2025 Jan-.

[0076] In some embodiments, the present disclosure provides compositions, methods and / or uses for treating disorders or other diseases associated with elevated red blood cells by administering a p2-specific adrenergic receptor antagonist. p2-specific adrenergic receptor antagonists include, for example, ICI-118551, butaxamine, and pharmaceutically acceptable salts of any of the foregoing, e.g., including, but not limited to, ICI-118551 hydrochloride and butaxamine hydrochloride.

[0077] In some embodiments, the present disclosure provides compositions, methods and / or uses for treating disorders or other diseases associated with elevated red blood cells by administering a pi / p2-specific adrenergic receptor antagonist. pi / p2-specific adrenergic receptor antagonists include, for example, propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, a pi / p2-specific adrenergic receptor antagonist is propranolol or a pharmaceutically acceptable salt thereof.ICI-118551

[0078] ICI-118551, also known as ICI-118,551, Zenidolol, or 3-(isopropylamino)-l- [(7-methyl-4-indanyl)oxy]butan-2-ol (CAS number 72795-26-7), is a selective P2 adrenergic receptor antagonist or beta blocker. ICI-118551 binds to the P2 subtype with at least 100times greater affinity than Pi or Ps, the two other known subtypes of the beta adrenoceptor. P2-AR is encoded by the ADRB2 gene. P2-AR are cell-membrane spanning receptors for adrenaline (epinephrine) that mediate smooth muscle relaxation and bronchodilation via adenylate cyclase stimulation (Abosamak NER, Shahin MH. Beta2 Receptor Agonists and Antagonists. [Updated 2023 Jul 3], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2024 Jan-; Johnson (2006) J Allergy Clin Immunol 117, 18-24; Yang et al. (2021) Life Sci 265: 118864), which are each incorporated herein by reference in their entirety.

[0079] ICI-118551 is well known in the art, being first described in EP0003664A1, published in 1979, which is incorporated herein by reference in its entirety. ICI-118551 is described in EP0003664A1 as a peripherally-selective beta-adrenergic blocking agents.

[0080] ICI-118551 has been characterized as an ocular hypotensive agent. Uathanson, J. A., ICI 118,551 : an effective ocular hypotensive agent with selectivity for the ciliary process beta 2-adrenoceptor and with minimal cardiac side effects. Br J Pharmacol. 1984 Nov;83(3):821-9, incorporated herein by reference in its entirety. ICI-118551 has also been found to have antiangiogenic properties in hemangioblastomas primary cell cultures. Cuesta, A.M., el al. The p2-adrenergic receptor antagonist ICI-118,551 blocks the constitutively activated H IF signalling in hemangiobl stomas from von Hippel-Lindau disease. Sci Rep 9, 10062 (2019), which is incorporated herein by reference in its entirety.

[0081] ICI-118551 has a molecular formula C17H27NO2. Exemplary molecular forms of ICI-118551 include, but are not limited to CAS reference 72795-26-7 and IUPAC name 3-(isopropylamino)-l-[(7-methyl-4-indanyl)oxy]butan-2-ol. ICI-118551 is or comprises a molecule corresponding to Formula I below.

[0082] Formula I

[0083] Commercially available forms of ICI-118551 include, without limitation, catalog number T68385 from TargetMol and catalog number HY- 100543 from MedChemExpress.

[0084] Example salt forms of ICI-118551 include, but are not limited to, ICI-118551 hydrochloride (molecular formula C17H28CIN2O2). Exemplary molecular forms of ICI- 118551 hydrochloride include, but are not limited to CAS reference 72795-01-8.Commercially available forms of ICI-118551 hydrochloride include, without limitation, catalog number HY-1395 Ifirom MedChemExpress, catalog number B 1004 from APExBIO, and catalog number S8114 from Selleckchem.

[0085] ICI-118551 compositions can be formulated, for example, as described in Remington's Pharmaceutical Sciences, 21st Edition, University of the Sciences in Philadelphia, Philadelphia, Pa., USA (2006).

[0086] In some embodiments, the disclosure provides herein new uses of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto, as described further herein.Butaxamine

[0087] Butaxamine, also known as Butoxamine, and a-(l-[tert-butylamino]ethyl)-2,5- dimethoxybenzyl alcohol, is a selective P2 adrenergic receptor antagonist or beta blocker that has predominantly been used in experimental situations.

[0088] Butaxamine has a molecular formula C15H25NO3. Example molecular forms of Butaxamine include, but are not limited to CAS reference 2922-20-5 and IUPAC name 2- (tert-butylamino)-l-(2,5-dimethoxyphenyl)propan-l-ol. Butaxamine is or comprises a molecule corresponding to Formula II below.

[0089] Formula II

[0090] Commercially available forms of butaxamine include, without limitation, catalog number Bl 0847395 from BenchChem and catalog number HY-119868 from MedChemExpress.

[0091] Exemplary salt forms of butaxamine include, but is not limited to, butaxamine hydrochloride (molecular formula C15H25NO3 HC1). Example molecular forms of butaxamine hydrochloride include, but are not limited to CAS reference 5696-15-1. Commercially available forms of butaxamine hydrochloride include, without limitation, catalog number HY-118470 from MedChemExpress, catalog number sc-234233 from Santa Cruz Biotechnology.

[0092] Butaxamine compositions can be formulated, for example, as described in Remington's Pharmaceutical Sciences, 21st Edition, University of the Sciences in Philadelphia, Philadelphia, Pa., USA (2006).

[0093] In some embodiments, the disclosure provides herein new uses of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto, as described further herein.Propranolol

[0094] Propranolol, also known as AY-64043, Bedranol, Beta-prograne, ICI-45520, NCS-91523, l-(isopropyl amino)-3-(l-naphthyloxy)-2-propanol, is a beta-adrenergic blocking agent. Propranolol is a competitive beta-adrenergic receptor antagonist that is devoid of agonist activity.

[0095] Propranolol is a nonselective beta-adrenoreceptor antagonist, also classified as a class II anti arrhythmic. It exerts its response by competitively blocking beta-1 and beta-2 adrenergic stimulation, e.g., in the heart, which is typically induced by epinephrine and norepinephrine. Shahrokhi M, Gupta V. Propranolol. [Updated 2023 May 1], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2024 Jan-, which is incorporated herein by reference in its entirety. Propranolol is commonly used for the treatment of a number of conditions including, for example, hypertension, heart failure, irregular heart rate, coronary artery disease, and essential tremors. Id.

[0096] Propranolol has a molecular formula C16H21NO2. Example molecular forms of propranolol include, but are not limited to CAS reference 525-66-6 and IUPAC name (RS)-l- (l-methylethylamino)-3-(l-naphthyloxy)propan-2-ol. Propranolol is or comprises a molecule corresponding to Formula III below.

[0098] Commercially available forms of propranolol include, without limitation, catalog number sc-507425 from Santa Cruz Biotechnology Inc. and catalog number FE-0204 from Key Organics.

[0099] Example salt forms of propranolol include, but are not limited to, propranolol hydrochloride (molecular formula C16H21NO2 HC1). Example molecular forms of propranolol hydrochloride include, but are not limited to CAS reference 318-98-9. Commercially available forms of propranolol hydrochloride include, without limitation, catalog number 23349 from Cayman Chemical, catalog number 1576005 from Millipore Sigma, catalog number HY-B0573 from MedChemExpress, and catalog number S4076 from Selleckchem.

[0100] Propranolol compositions can be formulated, for example, as described in Remington's Pharmaceutical Sciences, 21st Edition, University of the Sciences in Philadelphia, Philadelphia, Pa., USA (2006).

[0101] In some embodiments, the disclosure provides herein new uses of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto, as described further herein.Timolol

[0102] Timolol, also known as, Timoptic or Istalol, is a beta-adrenergic blocking agent that is typically used to treat conditions such as open-angle glaucoma, ocular hypertension, infantile hemangiomas, hypertension, myocardial infarction, migraine prevention, and atrial fibrillation. Barnes J, Moshirfar M. Timolol. [Updated 2024 Aug 17], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2025 Jan-.

[0103] Timolol has a molecular formula C13H24N4O3S. Example molecular forms of timolol include, but are not limited to CAS reference 26839-75-8 and IUPAC name (S)-l- (tert-Butylamino)-3-[(4-morpholin-4-yl-l,2,5-thiadiazol-3-yl)oxy]propan-2-ol. Timolol is a molecule corresponding to Formula IV below.

[0104] Formula IV

[0105] In some embodiments, the disclosure provides herein new uses of timolol or a pharmaceutically acceptable salt thereof for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto.Nadolol

[0106] Nadolol is a nonselective P-blocker (also referred to as a [31 / p2-specific P blocker) approved by the US Food and Drug Administration (FDA) for managing hypertension and angina. Nadolol also has been characterized as having various off-label uses, including the treatment of atrial fibrillation, ventricular arrhythmias linked to long QT syndrome, ventricular premature beats, catecholaminergic polymorphic ventricular tachycardia, supraventricular tachycardia, prophylaxis against gastroesophageal variceal hemorrhage in liver cirrhosis, thyrotoxicosis management, as well as migraine and vascular headache prophylaxis. Gopal S, Patel P, Mandiga P. Nadolol. [Updated 2024 Feb 22], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2025 Jan-.

[0107] Nadolol has a molecular formula C17H27NO4. Example molecular forms of nadolol include, but are not limited to CAS reference 42200-33-9 and IUPAC name (2R*,3S*)-5-{[(2R*)-3-(tert-butylamino)-2-hydroxypropyl]oxy}-l,2,3,4- tetrahydronaphthalene-2,3-diol. Nadolol is a molecule corresponding to Formula V below.

[0108] Formula V

[0109] In some embodiments, the disclosure provides herein new uses of nadolol or a pharmaceutically acceptable salt thereof for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto.Sotalol

[0110] Sotalol, also known as Betapace, Sorine, Sotylize, is a non-cardioselective P- blocker that possesses potassium channel-blocking properties. Sotalol is approved by the US Food and Drug Administration (FDA) and is indicated for treating hemodynamically stable ventricular tachycardia and maintaining sinus rhythm, particularly in cases of paroxysmal atrial fibrillation. Sotalol is also used as an off-label medication for pharmacological cardioversion of atrial fibrillation, premature ventricular contractions, postoperative atrial fibrillation after cardiac surgery, and supraventricular tachycardia (SVT). Mubarik A, Kerndt CC, Patel P, et al. Sotalol. [Updated 2024 Feb 26], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2025 Jan-.[OHl] Sotalol has a molecular formula C12H20N2O3S. Example molecular forms of sotalol include, but are not limited to CAS reference 3930-20-9 and IUPAC name (RS)-N-{4- [l-hydroxy-2-(propan-2-ylamino)ethyl]phenyl}methanesulfonamide. Sotalol is a molecule corresponding to Formula VI below.

[0112] Formula VI

[0113] In some embodiments, the disclosure provides herein new uses of sotalol or a pharmaceutically acceptable salt thereof for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto.Penbutolol

[0114] Penbutolol, also known as Levatol, is a nonselective beta-adrenergic receptor blocker (also referred to as a [31 / p2-specific P blocker) that is used to treat hypertension. Without wishing to be bound by theory, it is believed that the pi adrenergic blockade reduces the heart rate and myocardial contractility by slowing the AV conduction and suppressing automaticity and the P2 blockade affects peripheral vascular resistance and can cause bronchospasm and hypoglycemia. LiverTox: Clinical and Research Information on Drug- Induced Liver Injury [Internet], Bethesda (MD): National Institute of Diabetes and Digestive and Kidney Diseases; 2012-. Penbutolol. [Updated 2017 Jan 15],

[0115] Penbutolol has a molecular formula C18H29NO2. Example molecular forms of penbutolol include, but are not limited to CAS reference 38363-40-5 and IUPAC name (S)-l- (tert-butylamino)-3-(2-cyclopentylphenoxy)propan-2-ol. Penbutolol is a molecule corresponding to Formula VII below.

[0116] Formula VII

[0117] In some embodiments, the disclosure provides herein new uses of penbutolol or a pharmaceutically acceptable salt thereof for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto.Pindolol

[0118] Pindolol, also known as Visken, is a beta-adrenergic blocking agent. Pindolol is a nonselective beta-adrenergic receptor blocker (also referred to as a [31 / p2-specific P blocker) that is widely used for the therapy of hypertension and angina pectoris. LiverTox: Clinical and Research Information on Drug-Induced Liver Injury [Internet], Bethesda (MD): National Institute of Diabetes and Digestive and Kidney Diseases; 2012-. Pindolol. [Updated 2017 Jan 15],

[0119] Pindolol has a molecular formula C14H20N2O2. Example molecular forms of pindolol include, but are not limited to CAS reference 13523-86-9 and IUPAC name (RS)-1- (U7-indol-4-yloxy)-3-(isopropylamino)propan-2-ol. Pindolol is a molecule corresponding to Formula VIII below.

[0120] Formula VIII

[0121] In some embodiments, the disclosure provides herein new uses of pindolol or a pharmaceutically acceptable salt thereof for oral administration for treatment of a red blood cell disease or disorder, and / or condition related thereto.Treatment of red blood cell disorders

[0122] The present disclosure provides methods, compositions, and uses for the treatment of red blood cell diseases or disorders, and / or conditions related thereto. In some embodiments, a disease or disorder associated with elevated red blood cells is selected from a myeloproliferative disorder, myeloproliferative neoplasm, and secondary polycythemia.

[0123] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a 32-specific adrenergic receptor antagonist (e.g., ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are uses of 32-specific adrenergic receptor antagonists (e.g., ICI-118551, butaxamine, and pharmaceutically acceptable salts of any of the foregoing) in treating a red blood cell disease or disorder in a patient. In some embodiments, a p2-specific adrenergic receptor antagonist (e.g., ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0124] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in treating a red blood cell disease or disorder in a patient. In some embodiments, ICI- 118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0125] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride). In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in treating a red blood cell disease or disorder in a patient. In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0126] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are uses of pi / p2- specific adrenergic receptor antagonists (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, and pharmaceutically acceptable salts of any of the foregoing) in treating a red blood cell disease or disorder in a patient. In some embodiments, a pi / p2- specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0127] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride). In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in treating a red blood cell disease or disorder in a patient. In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0128] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of timolol or a pharmaceutically acceptable salt thereof. In some embodiments, provided are uses of timolol or a pharmaceutically acceptable salt thereof in treating a red blood cell disease or disorder in a patient. In some embodiments, timolol or a pharmaceutically acceptable salt thereof is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0129] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of nadolol or a pharmaceutically acceptable salt thereof. In some embodiments, provided are uses of nadolol or a pharmaceutically acceptable salt thereof in treating a red blood cell disease or disorder in a patient. In some embodiments,nadolol or a pharmaceutically acceptable salt thereof is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0130] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of sotalol or a pharmaceutically acceptable salt thereof. In some embodiments, provided are uses of sotalol or a pharmaceutically acceptable salt thereof in treating a red blood cell disease or disorder in a patient. In some embodiments, sotalol or a pharmaceutically acceptable salt thereof is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0131] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of penbutolol or a pharmaceutically acceptable salt thereof. In some embodiments, provided are uses of penbutolol or a pharmaceutically acceptable salt thereof in treating a red blood cell disease or disorder in a patient. In some embodiments, penbutolol or a pharmaceutically acceptable salt thereof is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0132] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of pindolol or a pharmaceutically acceptable salt thereof. In some embodiments, provided are uses of pindolol or a pharmaceutically acceptable salt thereof in treating a red blood cell disease or disorder in a patient. In some embodiments, pindolol or a pharmaceutically acceptable salt thereof is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder.

[0133] In some embodiments, a disease or disorder associated with elevated red blood cells is a myeloproliferative disorder. Myeloproliferative disorders are characterized as a group of diseases related to abnormal proliferation of blood cells produced in bone marrow. Myeloproliferative disorders include polycythemia vera (PV), essential thrombocythemia (ET), myelofibrosis (e.g., primary myelofibrosis, post-polycythemia vera myelofibrosis, and post-essential thrombocythemia myelofibrosis), chronic myeloid leukemia (CML). PV is characterized by increased production of all 3 types of blood cells, whereas ET is manifested in the elevation of platelets. In some embodiments, a myeloproliferative disorder is PV.

[0134] In some embodiments, provided herein are methods of treating PV in a patient in need thereof.

[0135] In some embodiments, provided herein are methods of treating PV in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a p2-specific adrenergic receptor antagonist (e.g., ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are uses of a p2-specific adrenergic receptor antagonist (e.g., ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) in treating PV in a patient. In some embodiments, a p2-specific adrenergic receptor antagonist (e.g., ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating PV.

[0136] In some embodiments, provided herein are methods of treating PV in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI- 118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in treating PV in a patient. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating PV.

[0137] In some embodiments, provided herein are methods of treating PV in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride). In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in treating PV in a patient. In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in the manufacture of a medicament for treating PV.

[0138] In some embodiments, provided herein are methods of treating PV in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are uses of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) in treating PV in a patient. In some embodiments, a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol,nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating PV.

[0139] In some embodiments, provided herein are methods of treating PV in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride). In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in treating PV in a patient. In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in the manufacture of a medicament for treating PV.

[0140] A common symptom of PV is high blood pressure. In some embodiments, provided herein are methods of treating a patient having PV and hypertension. In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering propranolol, or a pharmaceutically acceptable salt thereof, e.g. including, but not limited to, propranolol hydrochloride.

[0141] In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering timolol, or a pharmaceutically acceptable salt thereof. In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering nadolol, or a pharmaceutically acceptable salt thereof. In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering sotalol, or a pharmaceutically acceptable salt thereof. In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering penbutolol, or a pharmaceutically acceptable salt thereof. In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering pindolol, or a pharmaceutically acceptable salt thereof.

[0142] In some embodiments, provided methods, compositions, and uses are for the treatment of a red blood cell disease or disorder associated with JAK2 mutation. In some embodiments, provided methods, compositions, and uses are for the treatment of amyeloproliferative disorder associated with & .JAK2 mutation, wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, provided methods, compositions, and uses are for the treatment of PV associated with JAK2 mutation. In some embodiments, JAK2 mutation is Jak2V617F. Jak2V617F mutation is the most common molecular abnormality in BCR-ABL-negative myeloproliferative neoplasm (MPN) and is present in approximately 95% of patients with PV. Mullally et al. Physiological Jak2V617F expression causes a lethal myeloproliferative neoplasm with differential effects on hematopoietic stem and progenitor cells. Cancer Cell 2010 Jun 15;17(6):584-96, which is incorporated herein by reference in its entirety.

[0143] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a 32-specific adrenergic receptor antagonist (e.g., ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing), wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, provided are uses of a 32-specific adrenergic receptor antagonist (e.g., ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) in treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient, wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, a P2-specific adrenergic receptor antagonist (e.g., ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617k').iwherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia.

[0144] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride), wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient, wherein the JAK2 mutation is associated with elevated red bloodcell production, PV and / or polycythemia. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., ,Jak2V617k').iwherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia.

[0145] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride), wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient, wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617k').iwherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia.

[0146] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing), wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, provided are uses of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) in treating a red blood cell disease or disorder associated with a JAK2 mutation (e.g., Jak2V617F) in a patient, wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicamentfor treating a red blood cell disease or disorder associated with &.JAK2 mutation (e.g., Jak2V617F wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia.

[0147] In some embodiments, provided herein are methods of treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride), wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617F) in a patient, wherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in the manufacture of a medicament for treating a red blood cell disease or disorder associated with JAK2 mutation (e.g., Jak2V617k').iwherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia.

[0148] In some embodiments, provided methods, compositions, and uses are for the treatment of PV associated with JAK2 mutation. In some embodiments, the PV is associated with a Jak2V617F mutation.

[0149] In some embodiments, a disease or disorder associated with elevated red blood cells is a myeloproliferative disorder that has progressed to a cancer. In some embodiments, a disease or disorder associated with elevated red blood cells is a MPN.

[0150] In some embodiments, a disease or disorder associated with elevated red blood cells is secondary polycythemia. Secondary polycythemia is a heterogeneous group of disorders characterized by elevated red blood cells that can be due to a number of reasons, such as, for example, hypoxia, inappropriate erythropoietin secretion, secondary to other diseases and / or contributing factors. Haider MZ, Anwer F. Secondary Polycythemia.[Updated 2023 May 8], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2024 Jan-, which is incorporated by reference herein in its entirety.

[0151] In some embodiments, provided are methods for treating secondary polycythemia in a patient in need thereof.

[0152] In some embodiments, provided herein are methods of treating secondary polycythemia in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a p2-specific adrenergic receptor antagonist (e.g., ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are uses of a p2-specific adrenergic receptor antagonist (e.g., ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) in treating secondary polycythemia in a patient. In some embodiments, a P2-specific adrenergic receptor antagonist (e.g., ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating secondary polycythemia.

[0153] In some embodiments, provided herein are methods of treating secondary polycythemia in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in treating secondary polycythemia in a patient. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating secondary polycythemia.

[0154] In some embodiments, provided herein are methods of treating secondary polycythemia in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride). In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in treating secondary polycythemia in a patient. In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is provided herein for use in the manufacture of a medicament for treating secondary polycythemia.

[0155] In some embodiments, provided herein are methods of treating secondary polycythemia in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing). In some embodiments, provided are uses of a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) in treating secondarypolycythemia in a patient. In some embodiments, a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) is provided herein for use in the manufacture of a medicament for treating secondary polycythemia.

[0156] In some embodiments, provided herein are methods of treating secondary polycythemia in a patient in need thereof, the method comprising administering to the patient in need thereof an effective amount of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride). In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in treating secondary polycythemia in a patient. In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is provided herein for use in the manufacture of a medicament for treating secondary polycythemia.

[0157] In some embodiments, a secondary polycythemia is secondary to sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastoma and meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

[0158] In some embodiments, secondary polycythemia is the result of a central hypoxic process, chronic lung disease (e.g., COPD and Pickwickian syndrome), right-to-left cardiopulmonary vascular shunts, cyanotic heart disease, carbon monoxide poisoning, smoker’s erthrocytosis, hypoventilation syndromes (e.g., obstructive sleep apnea, obesity hypoventilation syndrome), high altitude habitat, and / or renal disease (e.g., local renal hypoxia, renal artery stenosis).

[0159] In some embodiments, secondary polycythemia is hemoglobinopathy with high-oxygen-affinity, decreased levels of erythrocyte 2, 3, -diphosphoglycerate (DPG), bi sphosphogly cerate mutase deficiency, methemoglobinemia, hereditary ATP increase, and / or an oxygen sensing pathway gene mutation (e.g., EpoR,3 VHL,8-10 and PHD216).

[0160] In some embodiments, a secondary polycythemia is associated with or caused by a tumor with excessive production of erythropoietin or erythropoietin related factors (e.g., renal cell carcinoma, hepatocellular carcinoma, pheochromocytoma, cerebellar hemangioblastoma, uterine leiomyoma, ovarian carcinoma, meningioma, parathyroid carcinoma / adenomas), erythropoietin administration, androgen administration, a renal disease(e.g., renal cysts, polycystic kidney disease, hydronephrosis, nephrotic syndrome, diffuse parenchymal disease, Bartter’s syndrome, end-stage renal disease, long-term hemodialysis, post-renal transplant erythrocytosis), adrenal cortical hypersecretion, and / or idiopathic polycythemia.

[0161] In some embodiments, secondary polycythemia is relative polycythemia. Relative polycythemia is an elevated hematocrit marked with a normal to high normal RBC mass and low normal to decreased plasma volume. In some embodiments, a relative polycythemia is Gaisbock syndrome, spurious, and / or stress erythrocytosis. In some embodiments, secondary polycythemia is Chuvash Polycythemia.

[0162] High blood pressure or hypertension is a risk factor for Secondary Polycythemia patients. In some embodiments, provided are methods for treating patients with both Secondary Polycythemia and hypertension. In some embodiments, provided are methods that treat both Secondary Polycythemia and hypertension by administering propranolol, or a pharmaceutically acceptable salt thereof, e.g. including, but not limited to, propranolol hydrochloride.

[0163] The present disclosure provides methods, compositions, and uses for the treatment of splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement in a patient with an associated red blood cell disease or disorder as described herein. In some embodiments, provided methods, compositions, and uses for the treatment of splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement in a patient with PV.

[0164] In some embodiments, provided herein are methods of treating splenomegaly in a patient with a red blood cell disorder as described herein, the method comprising administering to the patient in need thereof an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in treating splenomegaly in a patient with a red blood cell disorder as described herein. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating splenomegaly in a patient with a red blood cell disorder as described herein.

[0165] In some embodiments, provided herein are methods of normalizing splenic architecture in a patient with a red blood cell disorder as described herein, the methodcomprising administering to the patient in need thereof an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in normalizing splenic architecture in a patient with a red blood cell disorder as described herein. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for normalizing splenic architecture in a patient with a red blood cell disorder as described herein.

[0166] In some embodiments, provided herein are methods of treating splenic architecture effacement in a patient with a red blood cell disorder as described herein, the method comprising administering to the patient in need thereof an effective amount of ICI- 118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI- 118551 hydrochloride) is provided herein for use in treating splenic architecture effacement in a patient with a red blood cell disorder as described herein. In some embodiments, ICI- 118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating splenic architecture effacement in a patient with a red blood cell disorder as described herein.

[0167] In some embodiments, provided herein are methods of treating splenic erythroid hyperplasia in a patient with a red blood cell disorder as described herein, the method comprising administering to the patient in need thereof an effective amount of ICI- 118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride). In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI- 118551 hydrochloride) is provided herein for use in treating splenic erythroid hyperplasia in a patient with a red blood cell disorder as described herein. In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is provided herein for use in the manufacture of a medicament for treating splenic erythroid hyperplasia in a patient with a red blood cell disorder as described herein.Patients

[0168] A patient may be any human afflicted by a disease or disorder associated with elevated red blood cells. Methods and compositions encompassed by the present disclosurecan be used in treatment of patients with red blood cell diseases or disorders, and / or conditions related thereto, as described herein.

[0169] In some embodiments, terms “individual” or “patient” are used and are intended to be interchangeable with “subject.” A patient may be a human subject of any age. In some embodiments, a patient is an adult. In some embodiments, a patient is an adolescent. In some embodiments, a patient is a child. In some embodiments, a patient is an infant. In some embodiments, a patient may be an aging patient.

[0170] In some embodiments, a patient has a disease or disorder associated with elevated red blood cells is selected from a myeloproliferative disorder, myeloproliferative neoplasm, and secondary polycythemia.

[0171] In some embodiments, a patient has a myeloproliferative disorder associated with elevated red blood cells. In some embodiments, a patient has PV.

[0172] High blood pressure is a common symptom of PV patients. In some embodiments, a patient has PV and high blood pressure.

[0173] In some embodiments, a patient has a myeloproliferative disorder associated with JAK2 mutation (e.g., Jak2V617l'').iwherein the JAK2 mutation is associated with elevated red blood cell production, PV and / or polycythemia. In some embodiments, a patient has PV associated with a JAK2 mutation (e.g., Jak2V617F).

[0174] In some embodiments, a patient has a myeloproliferative neoplasm.

[0175] In some embodiments, a patient has secondary polycythemia.

[0176] In some embodiments, a patient has a secondary polycythemia that is secondary to sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastoma and meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

[0177] In some embodiments, a patient has a secondary polycythemia that is associated with a central hypoxic process, chronic lung disease (e.g., COPD and Pickwickian syndrome), right-to-left cardiopulmonary vascular shunts, cyanotic heart disease, carbon monoxide poisoning, smoker’s erthrocytosis, hypoventilation syndromes (e.g., obstructive sleep apnea, obesity hypoventilation syndrome), high altitude habitat, and / or renal disease (e.g., local renal hypoxia, renal artery stenosis), a hemoglobinopathy with high-oxygen-affinity, decreased levels of erythrocyte 2,3,-DPG, bi sphosphogly cerate mutase deficiency, methemoglobinemia, hereditary ATP increase, and / or an oxygen sensing pathway gene mutation (e.g., EpoR,3 VHL,8-10 and PHD216), a tumor with excessive production of erythropoietin or erythropoietin related factors (e.g., renal cell carcinoma, hepatocellular carcinoma, pheochromocytoma, cerebellar hemangioblastoma, uterine leiomyoma, ovarian carcinoma, meningioma, parathyroid carcinoma / adenomas), erythropoietin administration, androgen administration, a renal disease (e.g., renal cysts, polycystic kidney disease, hydronephrosis, nephrotic syndrome, diffuse parenchymal disease, Bartter’s syndrome, endstage renal disease, long-term hemodialysis, post-renal transplant erythrocytosis), adrenal cortical hypersecretion, idiopathic polycythemia, Chuvash Polycythemia, Gaisbock syndrome, spurious erythrocytosis, and / or stress erythrocytosis.

[0178] The patient may or may not have received previous treatment for the instant or other indications, such as chemotherapy (e.g., hydroxyurea), blood draws, or JAK2 inhibitors.

[0179] In some embodiments, a patient has a disease or disorder associated with elevated red blood cells (e.g., a myeloproliferative disorder, e.g., PV) and has previously received one or more prior therapies selected from chemotherapy (e.g., hydroxyurea), blood draws, and treatment with a JAK2 inhibitor. In some embodiments, a patient has a disease or disorder associated with elevated red blood cells (e.g., a myeloproliferative disorder, e.g., PV) and treatment with a pi / p2-specific adrenergic receptor antagonist or a p2-specific adrenergic receptor antagonist is a first-line therapy for the disease or disorder.

[0180] The present disclosure provides methods, compositions, and uses for the treatment of splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement in a patient with an associated red blood cell disease or disorder as described herein. In some embodiments, provided methods, compositions, and uses for the treatment of splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement in a patient with PV.

[0181] In some embodiments, a patient has a disease or disorder associated with elevated red blood cells and also has splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement. In some embodiments, a patient has (i) a myeloproliferative disorder and (ii) splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement. In some embodiments, a patient has (i) PV and (ii) splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement.

[0182] In some embodiments, a patient for treatment by methods described herein has elevated red blood cell mass. A determination of an elevated red blood cell mass is within the scope of medical judgment. For example, in some embodiments, a normal healthy adult, red blood cell mass is 23 to 29 mL / kg in females and 26 to 32 mL / kg in males. In some embodiments, a patient for treatment by methods described herein is a female patient with a red blood mass greater than 29 mL / kg or a male patient with a red blood mass greater than 32 mL / kg.

[0183] In some embodiments, a patient for treatment by methods described herein has elevated hematocrit. A determination of an elevated hematocrit value is within the scope of medical judgment. In some embodiments, a patient for treatment by methods described herein is a female patient with a hematocrit value greater than 48% or a male patient with a hematocrit value greater than 51%.

[0184] In some embodiments, a patient for treatment by methods described herein has elevated hemoglobin. A determination of an elevated hemoglobin value is within the scope of medical judgment. In some embodiments, a patient for treatment by methods described herein is a female patient with a hemoglobin value greater than 165 g / L or a male patient with a hemoglobin value greater than 185g / L.

[0185] In some embodiments, a patient for treatment by methods described herein has a red blood cell mass greater than 125% of the predicted value adjusted for gender and body weight.

[0186] In some embodiments, a patient has an increased blood viscosity.Administration of Agents

[0187] P2 adrenergic receptor antagonists, such as, for example, ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing (e.g., ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride) can be administered to patients disclosed herein according to methods encompassed by the present disclosure in a biologically compatible form suitable for pharmaceutical administration in vivo. By “biologically compatible form suitable for administration in vivo" is meant a form to be administered in which any toxic effects are outweighed by the therapeutic effects. Administration of a P2 adrenergic receptor antagonist, such as, ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing (e.g., ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride) can be in anypharmacological form including a therapeutically active amount of an agent alone or in combination with a pharmaceutically acceptable carrier.

[0188] Therapeutic agents as described herein, such as P2 adrenergic receptor antagonists (e.g., ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing, e.g., ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride) can be administered in a convenient manner such as by oral administration, injection (subcutaneous, intravenous, intraperitoneal (i.p.) etc.), inhalation, transdermal application, or rectal administration. Depending on the route of administration, the active compound can be coated in a material to protect the compound from the action of enzymes, acids and other natural conditions which can inactivate the compound. For example, for administration of agents, by other than parenteral administration, it can be desirable to coat the agent with, or co-administer the agent with, a material to prevent its inactivation.

[0189] As described in detail below, the pharmaceutical compositions encompassed by the present disclosure (e.g., a composition comprising a P2 adrenergic receptor antagonist, e.g., ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing, such as, e.g., ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride) can be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, boluses, powders, granules, pastes; or (2) parenteral administration, for example, by subcutaneous, intramuscular, intraperitoneal (i.p.) or intravenous injection as, for example, a sterile solution or suspension.

[0190] The phrase “pharmaceutically acceptable” is employed herein to refer to those agents, 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 without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0191] The phrase “pharmaceutically-acceptable carrier” as used herein means 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 chemical 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 and not injurious to the patient. Those skilled in the art will recognize, orbe able to employ suitable pharmaceutically-acceptable carriers for the therapeutic agent disclosed herein (e.g., a P2 adrenergic receptor antagonist, e.g., ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing, such as, e.g., ICI- 118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride) from the pharmaceutically-acceptable carriers known in pharmaceutical science (See, for example, Adejare, Adeboye, ed. Remington: the science and practice of pharmacy. Academic Press, 2020).

[0192] The term “pharmaceutically-acceptable salts” refers to the relatively nontoxic, inorganic and organic acid addition salts of the therapeutic agents disclosed herein (e.g., a P2 adrenergic receptor antagonist, e.g., ICI-118551, butaxamine, and propranolol, the pharmaceutically-acceptable salts of which include, for example, ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride, respectively). These salts can be prepared in situ during the final isolation and purification of the therapeutic agents, or by separately reacting a purified therapeutic agent in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Those skilled in the art will recognize, or be able to employ suitable pharmaceutically-acceptable salts for the therapeutic agents disclosed herein (e.g., P2 adrenergic receptor antagonist, e.g., ICI-118551, butaxamine, and propranolol, the pharmaceutically-acceptable salts of which include, for example, ICI-118551 hydrochloride, butaxamine hydrochloride, propranolol hydrochloride, respectively) from the pharmaceutically-acceptable salts known in pharmaceutical science (See, for example, Berge et al. (1977) J. Pharm. Sci. 66: 1-19).

[0193] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0194] Examples of pharmaceutically-acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0195] Formulations useful in the methods encompassed by the present disclosure include those suitable for oral administration, intravenous administration and / or administration by injection (e.g., intraperitoneal (i.p.) injection). The formulations can conveniently be presented in unit dosage form and can be prepared by any methods well- known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration.

[0196] In some embodiments, a 32-specific adrenergic receptor antagonist (e.g., ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing) is in a form formulated for oral administration.

[0197] In some embodiments, ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride) is in a form formulated for oral administration. Exemplary oral forms of ICI-118551 can be found in the art. In some embodiments, the agent provided herein is an oral formulation of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride).

[0198] In some embodiments, butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride) is in a form formulated for oral administration. In some embodiments, the agent provided herein is an oral formulation of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride).

[0199] In some embodiments, a pi / p2-specific adrenergic receptor antagonist (e.g., propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing) is in a form formulated for oral administration.

[0200] In some embodiments, propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride) is in a form formulated for oral administration. Exemplary oral forms of propranolol can be found in the art. In some embodiments, the agent provided herein is an oral formulation of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride).

[0201] In general, the formulations are prepared by uniformly and intimately bringing into association a therapeutic agent with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0202] Formulations suitable for oral administration can be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a therapeutic agent(s) as an active ingredient(s). A compound can also be administered as a bolus, electuary or paste.

[0203] In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, acetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions can also comprise buffering agents. Solid compositions of a similar type can also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0204] A tablet can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfaceactive or dispersing agent. Molded tablets can be made by molding in a suitable machine a mixture of the powdered peptide or peptidomimetic moistened with an inert liquid diluent.

[0205] Tablets, and other solid dosage forms, such as dragees, capsules, pills and granules, can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well-known in the pharmaceutical-formulating art. They can also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They can besterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions, which can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions can also optionally contain opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions, which can be used include polymeric substances and waxes. The active ingredient(s) can also be in micro- encapsulated form, if appropriate, with one or more of the above-described excipients.

[0206] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient(s), the liquid dosage forms can contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, com, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0207] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0208] Suspensions, in addition to the active agent can contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0209] Pharmaceutical compositions encompassed by the present disclosure suitable for parenteral administration comprise one or more therapeutic agents in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which can be reconstituted into sterile injectable solutions or dispersions just prior to use, which can contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0210] These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents, for example,paraben, chlorobutanol, phenol sorbic acid, and the like. It can also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0211] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, can depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0212] Injectable depot forms are made by forming microencapsule matrices of therapeutic agent(s) (e.g., ICI-118551, butaxamine, propranolol, or a pharmaceutically acceptable salt of any of the foregoing), in biodegradable polymers such as polylactidepolyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissue.

[0213] When the therapeutic agents encompassed by the present disclosure are administered as pharmaceuticals, to humans, they can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

[0214] It will be understood that the total daily dosage of the compounds of the present disclosure will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disease or disorder associated with elevated red blood cells being treated and the severity thereof; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and plasma half-life of the specific compound employed; the duration of the treatment; drugs used conjointly or coincidental with the specific antagonist employed; and like factors well known in the medical arts.

[0215] In some embodiments, the daily dosage of the active ingredient may include, but is not limited to, about 0.1 pg / kg to 100 mg / kg per adult per day. In some embodiments, the pharmaceutical compositions contain 0.1 pg / kg, 0.5 pg / kg, 1 pg / kg, 5 pg / kg, 10 pg / kg, 0.1 mg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, or 100 mg / kg of ICI- 118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride).

[0216] In some embodiments, the daily dosage of the active ingredient may include, but is not limited to, about 0.01 mg to 2000 mg per adult per day. In some embodiments, the pharmaceutical compositions contain 0.01 mg, 0.1 mg, 1 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 750 mg, 1000 mg, 1250 mg, 1500 mg, 1750 mg, or 2000 mg of ICI-118551 or a pharmaceutically acceptable salt thereof (e.g., ICI-118551 hydrochloride).

[0217] In some embodiments, the daily dosage of the active ingredient may include, but is not limited to, about 0.1 pg / kg to 100 mg / kg per adult per day. In some embodiments, the pharmaceutical compositions contain 0.1 pg / kg, 0.5 pg / kg, 1 pg / kg, 5 pg / kg, 10 pg / kg, 0.1 mg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, or 100 mg / kg of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride).

[0218] In some embodiments, the daily dosage of the active ingredient may include, but is not limited to, about 0.01 mg to 2000 mg per adult per day. In some embodiments, the pharmaceutical compositions contain 0.01 mg, 0.1 mg, 1 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 750 mg, 1000 mg, 1250 mg, 1500 mg, 1750 mg, or 2000 mg of butaxamine or a pharmaceutically acceptable salt thereof (e.g., butaxamine hydrochloride).

[0219] In some embodiments, the daily dosage of the active ingredient may include, but is not limited to, about 0.1 pg / kg to 100 mg / kg per adult per day. In some embodiments, the pharmaceutical compositions contain 0.1 pg / kg, 0.5 pg / kg, 1 pg / kg, 5 pg / kg, 10 pg / kg, 0.1 mg / kg, 1 mg / kg, 5 mg / kg, 10 mg / kg, 12.5 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, or 100 mg / kg of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride).

[0220] In some embodiments, the daily dosage of the active ingredient may include, but is not limited to, about 0.01 mg to 2000 mg per adult per day. In some embodiments, the pharmaceutical compositions contain 0.01 mg, 0.1 mg, 1 mg, 5 mg, 10 mg, 12.5 mg, 15 mg,20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 750 mg, 1000 mg, 1250 mg, 1500 mg, 1750 mg, or 2000 mg of propranolol or a pharmaceutically acceptable salt thereof (e.g., propranolol hydrochloride).

[0221] Actual dosage levels of the active ingredients in pharmaceutical compositions encompassed by the present disclosure can be determined by the methods encompassed by the present disclosure to obtain an amount of the active ingredient, which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.Numbered Illustrative Embodiments

[0222] Embodiment 1. A method of treating a disease or disorder associated with elevated red blood cells in a patient in need thereof, the method comprising administering to the patient an effective amount of a P2 adrenergic receptor antagonist that is ICI-118551 or a pharmaceutically acceptable salt thereof.

[0223] Embodiment 2. A method of treating a disease or disorder associated with elevated red blood cells in a patient in need thereof, the method comprising administering to the patient an effective amount of a P2 adrenergic receptor antagonist that is butaxamine or a pharmaceutically acceptable salt thereof.

[0224] Embodiment 3. A method of treating a disease or disorder associated with elevated red blood cells in a patient in need thereof, the method comprising administering to the patient an effective amount of a P2 adrenergic receptor antagonist that is propranolol or a pharmaceutically acceptable salt thereof.

[0225] Embodiment 4. The method of any one of embodiments 1 to 3, wherein the disease or disorder associated with elevated red blood cells is a myeloproliferative disorder.

[0226] Embodiment 5. The method of embodiment 4, wherein the myeloproliferative disorder is polycythemia vera (PV).

[0227] Embodiment 6. The method of embodiment 4 or embodiment 5, wherein the patient has a myeloproliferative disorder associated with a JAK2 mutation.

[0228] Embodiment 7. The method of embodiment 6, wherein the JAK2 mutation is a Jak2V617F mutation.

[0229] Embodiment 8. The method of any one of embodiments 4 to 7, wherein the myeloproliferative disorder has progressed to a cancer.

[0230] Embodiment 9. The method of embodiment 1, wherein the disease or disorder associated with elevated red blood cells is secondary polycythemia.

[0231] Embodiment 10. The method of embodiment 9, wherein the secondary polycythemia is caused by sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastoma and meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

[0232] Embodiment 11. The method of any one of embodiments 1 and 4 to 10, wherein the patient has splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement.

[0233] Embodiment 12. The method of embodiment 11, wherein the administering to the patient an effective amount of the P2 adrenergic receptor antagonist treats the splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement.

[0234] Embodiment 13. A method of decreasing red blood cells in a patient, the method comprising administering to the patient an effective amount of a P2 adrenergic receptor antagonist selected from ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

[0235] Embodiment 14. A method of decreasing erythroid differentiation in a patient, the method comprising administering to the patient an effective amount of a P2 adrenergic receptor antagonist selected from ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

[0236] Embodiment 15. A method of treating elevated red blood cells in a patient, the method comprising administering to the patient an effective amount of a P2 adrenergic receptor antagonist selected from ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

[0237] Embodiment 16. A method of treating splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement in a patient, the method comprising administering to the patient an effective amount of ICI-118551 or a pharmaceutically acceptable salt thereof.

[0238] Embodiment 17. The method of any one of embodiments 13 to 16, wherein the patient has a disease or disorder associated with elevated red blood cells.

[0239] Embodiment 18. The method of embodiment 17, wherein the disease or disorder associated with elevated red blood cells is a myeloproliferative disorder.

[0240] Embodiment 19. The method of embodiment 18, wherein the myeloproliferative disorder is polycythemia vera (PV).

[0241] Embodiment 20. The method of embodiment 18 or embodiment 19, wherein the patient has a myeloproliferative disorder associated with a JAK2 mutation.

[0242] Embodiment 21. The method of embodiment 20, wherein the JAK2 mutation is a Jak2V617F mutation.

[0243] Embodiment 22. The method of any one of embodiments 18 to 21, wherein the myeloproliferative disorder has progressed to a cancer.

[0244] Embodiment 23. The method of embodiment 17, wherein the disease or disorder associated with elevated red blood cells is secondary polycythemia.

[0245] Embodiment 24. The method of embodiment 23, wherein the secondary polycythemia is caused by sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastoma and meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

[0246] Embodiment 25. The method of any one of embodiments 1 to 24, wherein the method does not induce anemia in the patient.

[0247] Embodiment 26. The method of any one of embodiments 1 to 25, wherein the P2 adrenergic receptor antagonist is orally administered to the patient.

[0248] Embodiment 27. The method of any one of embodiments 1 and 4 to 26, wherein the P2 adrenergic receptor antagonist is ICI-118551.

[0249] Embodiment 28. The method of any one of embodiments 1 and 4 to 26, wherein the P2 adrenergic receptor antagonist is ICI-118551 hydrochloride.

[0250] Embodiment 29. The method of any one of embodiments 2, 4 to 10, 13 to15, and 17 to 26, wherein the P2 adrenergic receptor antagonist is butaxamine.

[0251] Embodiment 30. The method of any one of embodiments 2, 4 to 10, 13 to15, and 17 to 26, wherein the P2 adrenergic receptor antagonist is butaxamine hydrochloride.

[0252] Embodiment 31. The method of any one of embodiments 3 to 10, 13 to15, and 17 to 26, wherein the P2 adrenergic receptor antagonist is propranolol.

[0253] Embodiment 32. The method of any one of embodiments 3 to 10, 13 to 15, and 17 to 26, wherein the P2 adrenergic receptor antagonist is propranolol hydrochloride.

[0254] Embodiment 33. The method of any one of embodiments 1 to 32, wherein the patient is a human.

[0255] Embodiment 34. A pharmaceutical composition comprising ICI-118551 or a pharmaceutically acceptable salt thereof for use in a method of any one of embodiments 1 and 4 to 28 and 33.

[0256] Embodiment 35. Use of ICI-118551 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease or disorder associated with elevated red blood cells.

[0257] Embodiment 36. The use of embodiment 35, wherein the medicament further treats splenomegaly, splenic erythroid hyperplasia and / or splenic architecture effacement.

[0258] Embodiment 37. A pharmaceutical composition comprising butaxamine or a pharmaceutically acceptable salt thereof for use in a method of any one of embodiments 2, 4 to 10, 13 to 15, and 17 to 26, 29 to 30, and 33.

[0259] Embodiment 38. Use of butaxamine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease or disorder associated with elevated red blood cells.

[0260] Embodiment 39. A pharmaceutical composition comprising propranolol or a pharmaceutically acceptable salt thereof for use in a method of any one of embodiments 3 to 10, 13 to 15, and 17 to 26 and 31 to 33.

[0261] Embodiment 40. Use of propranolol or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease or disorder associated with elevated red blood cells.EXAMPLES

[0262] Described herein is a therapeutic approach of targeting red blood cell disorders using the P? adrenergic receptor antagonists or beta blockers ICI-118551, butaxamine, and propranolol. Administration of ICI-118551, butaxamine, or propranolol, each significantly dampens erythroid progenitors and RBC parameters in wild-type mice, wild-type mice treated with erythropoietin, and Polycythemia Vera model mice carrying a Jak2V617Fmutation in their hematopoietic compartment. ICI-118551 was also found to reduce splenomegaly and improve splenic architecture in Polycythemia Vera model mice.Example 1: ICI-118551 and propranolol decrease erythropoiesis in wild-type mice in vivo

[0263] The present example describes the unexpected discovery that ICI-118551, a selective P2 adrenergic receptor antagonist or beta blocker, decreases erythropoiesis.

[0264] The possible effects of a selective P2 adrenergic receptor antagonist ICI- 118551 and a non-selective P-adrenergic receptor antagonist, propranolol, were assessed in wild-type cells. Specifically, ICI-118551 (Selleckchem S8114) and propranolol (Selleckchem S4076) were administered to human donor-derived HSPCs (Representative from n=5 donors, n=4 technical replicates within donor) at doses of 1 pM and 5 pM. As shown in FIG. 1A administration of ICI-118551 (ICI) or propranolol to primary human HSPCs and dampens erythroid differentiation (CD235a), dose-dependently. As shown in FIG. IB, both ICI- 118551 and propranolol do not impact megakaryopoiesis (CD41 / 61 staining). These results show that ICI-118551 more potently inhibits erythropoiesis than propranolol.

[0265] The effects of ICI-118551 treatment on red blood cell parameters was investigated in wild-type mice. ICI-118551 treatment was administered according to the schematic shown in FIG. 2. Specifically, ICI-118551 treatment was by intraperitoneal (i.p.) administration in wild-type mice at 0.5 mg / kg for 30 days daily. Cheek bleeds were performed on days -4, 7, 14, 21, and 30, and complete blood count (CBC) profiling performed. Cheek bleeds may cause an initial reduction in red blood cell parameters in wildtype mice until a steady state is reached. Resulting effects on red blood cell parameters are shown in FIG. 3A-FIG. 3C. ICI-118551 treatment decreases RBC parameters in wild-type mice at steady state. FIG. 3A shows a decrease in RBCs, FIG. 3B shows a decrease in hematocrit (HCT)%, and FIG. 3C shows a decrease in hemoglobin (Hb) in the peripheral blood of wild-type mice treated with ICI-118551.

[0266] This same treatment was not shown to have an effect on the white blood cell parameters or platelets in wild-type mice at steady state. As shown in FIG. 4A-FIG. 4D, ICI- 118551 treatment does not significantly affect WBCs (FIG. 4A), platelets (FIG. 4B), monocytes (FIG. 4C), or lymphocytes (FIG. 4D) in the peripheral blood of ICI-118551- treated wild-type mice at steady state.

[0267] ICI-118551 treatment of wild-type mice were found to decrease red blood cell parameters at steady state, but does not lead to anemia in wild-type mice. Daily ICI-118551 treatment via i.p. administration for 30 days mildly decreases hemoglobin (Hb) (FIG. 5A) and hematocrit (HCT)% (FIG. 5B) in the peripheral blood after 30 days of daily i.p. administration, but does not lead to anemia.

[0268] These data support that ICI-118551 can specifically decrease erythropoiesis, without leading to anemia.Example 2: ICI-118551 decreases erythropoiesis in EPO-treated mice

[0269] The effect of ICI-118551 on erythropoiesis was investigated in mice with treated with Erythropoietin (EPO). EPO treatment of mice was at a dose of 500 lU / kg 3x weekly administered i.p.; 5 female mice per group were evaluated. Results are shown in FIG. 6A and FIG. 6B. As expected, an increase in Hemoglobin (Hb) and Hematocrit (HCT) was observed in the peripheral blood of mice treated with EPO and vehicle (DMSO). ICI treatment 3x weekly i.p. administration normalizes RBC parameters in Erythropoietin (EPO) treated mice. ICI treatment 0.5 mg / kg decreases (FIG. 6A) Hemoglobin (Hb) and (FIG. 6B) Hematocrit (HCT) in the peripheral blood of mice treated with EPO.

[0270] It is noted that the EPO concentration used is roughly -6.66 lU / ml blood for mice (secondary polycythemia patients have an average EPO concentration of 23.7 mIU / ml blood). Since ICI normalized the impact of EPO even at these relatively high doses it should hold promise for secondary polycythemia patients.

[0271] These data support that ICI-118551 can specifically decrease erythropoiesis in red blood cell disorders associated with elevated EPO.Example 3: ICI-118551 decreases erythropoiesis in PV-model mice

[0272] The effect of ICI-118551 on erythropoiesis was investigated in Polycythemia Vera (PV) model mice, carrying Jak2V617F mutation in their hematopoietic compartment. Mullally et al., supra. It is known that PV patients (containing Jak2V617 mutation) usually have subnormal or low EPO levels, while still having elevated red blood cells. Thus, the PV model (Jak2V617F mutant) mice serve as a more representative mouse model for this indication.

[0273] PV model and littermate control mice were treated with ICI-118551 at 0.5 mg / kg ICI at 5x weekly dosing for 6 weeks by i.p. administration. It was found that ICI- 118551 normalizes RBC parameters in PV model mice. FIG. 7 shows that ICI-118551treatment normalizes Hemoglobin (Hb) in the peripheral blood of PV model mice carrying Jak2V617F mutation in their hematopoietic compartment. It is noted that two out of five mice from the PV+dm group spontaneously died and thus were not included in the data shown. No PV+ici (n=5), LC+ici (n=6), or LC+dm (n=6) mice spontaneously died over the course of the experiment.

[0274] ICI-118551 treatment dampens erythroid progenitors in the bone marrow (BM) and spleen of PV-mouse model (Jak2V617F mutant mice). As shown in FIG. 8A-FIG. 8C, ICI-118551 treatment 5x weekly for 6 weeks i.p. decreases erythroid progenitors in the bone marrow (BM) and spleen of PV-model mice. ICI treatment 0.5 mg / kg 5x weekly i.p. administration reduces erythroid progenitors in the BM (FIG. 8A), colony-forming unit- erythroid (CFU-e) in the BM (FIG. 8B), and erythroid progenitors in the spleen (FIG. 8C) of PV model mice.

[0275] These data support that ICI-118551 can decrease erythropoiesis in PV subjects.Example 4: ICI-118551 normalizes splenomegaly and splenic architecture in PV-model mice

[0276] The effect of ICI-118551 on splenomegaly and splenic architecture effacement in PV-model mice was investigated. Specifically, PV-model and littermate controls were treated with ICI-118551 at 0.5 mg / kg 5x weekly i.p. administration for 6 weeks. ICI-118551 treatment was found to reduce spleen size (FIG. 9A) and spleen weight (FIG. 9B) in PV model mice. This treatment was also found to normalize splenic architecture in PV model mice. As shown in FIG. 10, histologic H&E stains showing erythroid hyperplasia and complete effacement of splenic architecture in PV mice, and improvement towards normalcy in ICI-118551 -treated PV mice.

[0277] These data support that ICI-118551 also has a beneficial effect on spleen size and splenic architecture in PV subjects.Example 5: Butaxamine inhibits erythroid differentiation of human HSPCs in vitro

[0278] The possible effects of the P2 adrenergic receptor antagonist butaxamine (Buta) was assessed in human donor-derived hematopoietic stem and progenitor cells (HSPCs).

[0279] Specifically, Buta (MedChem Express HY-118470) was administered at doses of 1 pM, 2 pM and 5 pM to primary human HSPCs. As shown in FIG. 11 A, administrationof Buta to primary human HSPCs at 2 pM and 5 pM doses potently dampens erythroid differentiation (CD235a), but Buta does not impact myelopoiesis (CD1 lb staining) (FIG. 11B) or megakaryopoiesis (CD41 / 61 staining)(FIG. 11C). n=4 technical replicates within each donor.Example 6: Butaxamine decreases erythropoiesis in EPO-treated mice

[0280] The effect of butaxamine on erythropoiesis was investigated in mice treated with Erythropoietin (EPO). EPO treatment of mice was at a dose of 500 lU / kg 3x weekly administered i.p.; 5 mice per group were evaluated. A schematic of the experimental plan is shown in FIG. 12. Results are shown in FIG. 13A and FIG. 13B. Buta treatment 3x weekly i.p. administration normalizes RBC parameters in Erythropoietin (EPO) treated mice. Buta treatment 0.5 mg / kg decreases (FIG. 13 A) red blood cells (RBCs) and (FIG. 13B) Hemoglobin (Hb) in the peripheral blood of mice treated with EPO.Example 7: Butaxamine effect on body weight in wild-type and EPO-treated mice

[0281] The effect of butaxamine on body weight was investigated in wild-type mice and mice treated with Erythropoietin (EPO) as a model of secondary polycythemia. Body weights were evaluated over a period of 40 days; results are shown in FIG. 14. There was no significant increase or decrease in body weights observed between wild-type or EPO-treated mice, and the body weights were not altered in these mice when treated with Buta.Example 8: Butaxamine decreases erythropoiesis in PV-model mice

[0282] The effect of butaxamine on erythropoiesis was investigated in Polycythemia Vera (PV) model mice, carrying Jak2V617F mutation in their hematopoietic compartment. Mullally etal., supra. As noted above, it is known that PV patients (containing Jak2V617 mutation) usually have subnormal or low EPO levels, while still having elevated red blood cells. Thus, Jak2V617F mutant mice serve as a representative mouse model for PV.

[0283] PV model and littermate control mice were treated with Buta at 0.5 mg / kg Buta or Vehicle (DMSO) at 5x weekly dosing by i.p. administration. An experimental plan is shown in FIG. 15. It was found that Buta normalizes RBC parameters in PV model mice. FIG. 16, FIG. 17A, and FIG. 17B show that Buta treatment normalizes red blood cell (RBC) parameters in the peripheral blood of PV model mice carrying Jak2V617F mutation in their hematopoietic compartment. Specifically, FIG. 16 shows that Buta treatment normalizes Hb, FIG. 17A shows that Buta treatment normalizes RBCs, and FIG. 17B shows that Buta treatment normalizes hematocrit (HCT).

[0284] Buta treatment also decreases erythroid progenitors in the bone marrow (BM) and spleen of PV-mouse model (Jak2V617F mutant mice). As shown in FIG. 18A-FIG. 18B, Buta treatment 5x weekly for 4 weeks i.p. decreases erythroid progenitors in the bone marrow (BM) of PV-model mice. Buta treatment 0.5 mg / kg 5x weekly i.p. administration reduces erythroid progenitors in the bone marrow (BM) (FIG. 18A), colony-forming unit-erythroid (CFU-e) in the BM (FIG. 18B) of PV model mice. FIG. 19 shows that Buta treatment 5x weekly for 4 weeks i.p. decreases erythroid progenitors in the spleen of PV model mice.

[0285] Buta treatment does not increase or decrease myelopoiesis or megakaryopoiesis in the BM of PV-model mice (Jak2V617F mutant mice). Buta treatment did not increase or decrease granulocytic monocyte progenitors (GMP) or megakaryocytic progenitors (mkP) in the BM of PV-model mice, as shown in FIG. 20A and FIG. 20B, respectively.

[0286] These data support that Buta can decrease erythropoiesis in PV subjects.Example 9: Propranolol treatment of PV-model mice

[0287] The effect of propranolol on erythropoiesis was investigated in Polycythemia Vera (PV) model mice, carrying Jak2V617F mutation in their hematopoietic compartment. Mullally etal., supra. As noted above, it is known that PV patients (containing Jak2V617 mutation) usually have subnormal or low EPO levels, while still having elevated red blood cells. Thus, Jak2V617F mutant mice serve as a representative mouse model for PV.

[0288] PV model mice were treated with propranolol at 0.3 mg / kg at 5x weekly dosing for 4 weeks by i.p. administration. It was found that propranolol normalizes Hemoglobin (Hb) in PV model mice, as shown in FIG. 21. While the normalization of Hb in propranolol treated mice was not quite statistically significant, this is believed to be due to the small experimental sample size. Overall, these data support that propranolol can decrease red blood cell parameters in PV subjects.Example 10: Comparison of ICI-118551, butaxamine, and propranolol treatment on erythropoiesis in EPO-treated mice

[0289] The effect of ICI-118551, butaxamine, and propranolol on erythropoiesis was investigated in mice treated with Erythropoietin (EPO). EPO treatment of mice was at a dose of 500 lU / kg 3x weekly administered i.p.; 5 mice per group were evaluated. A schematic of the experimental plan is shown in FIG. 22.

[0290] ICI 118551, Butaxamine, and Propranolol were all found to normalize RBC parameters in EPO-induced mouse model of secondary Polycythemia. Specifically, ICI, Buta, and Pro treatments 0.5 mg / kg 3xweekly i.p. decreases RBCs (FIG. 23A) and Hematocrit (HCT; FIG. 23B) in the peripheral blood of mice treated with Erythropoietin (EPO). Moreover, ICI, Buta and Pro showed similar potency in their effects on both RBCs and HCT. ICI, Buta, and Pro also normalize hemoglobin (Hb) in EPO-induced mouse model of secondary Polycythemia. As shown in FIG. 24A and FIG. 24B, ICI, Buta, and Pro treatments 0.5 mg / kg all decrease hemoglobin (Hb) in the peripheral blood of mice treated with Erythropoietin (EPO).

[0291] ICI, Buta, and Pro were also shown to not affect WBCs or platelets in EPO- induced mouse model of secondary Polycythemia. As shown in FIG. 25A and FIG. 25B, ICI, Buta, and Pro treatments 0.5 mg / kg 3x weekly i.p. do not affect WBCs or platelets in the peripheral blood of mice treated with Erythropoietin (EPO).

[0292] These effects on erythropoiesis were not associated with any change in body weight for each of ICI-118551, butaxamine and propranolol. Body weights were assessed in ICI- 118551, butaxamine, and propranolol treated (0.5 mg / kg 3x weekly i.p.) EPO-induced mouse model of secondary Polycythemia (EPO: 500IU / kg 3x weekly i.p. dose). The percent body weight change after 30 days was not significant between all tested groups (data not shown).

[0293] These data support that ICI-118551, butaxamine, and propranolol can each decrease erythropoiesis in red blood cell disorders associated with elevated EPO, without leading to anemia.INCORPORATION BY REFERENCE

[0294] All publications, patents, and patent applications mentioned herein are hereby incorporated by reference in their entirety as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.EQUIVALENTS

[0295] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

CLAIMSWhat is claimed is:

1. A method of treating a disease or disorder associated with elevated red blood cells in a patient in need thereof, the method comprising administering to the patient an effective amount of a 32-specific adrenergic receptor antagonist.

2. The method of claim 1, wherein the 32-specific adrenergic receptor antagonist is ICI- 118551 or a pharmaceutically acceptable salt thereof.

3. The method of claim 1, wherein the p2-specific adrenergic receptor antagonist is butaxamine or a pharmaceutically acceptable salt thereof.

4. A method of treating a disease or disorder associated with elevated red blood cells in a patient in need thereof, the method comprising administering to the patient an effective amount of a pi / p2-specific adrenergic receptor antagonist.

5. The method of claim 4, wherein the pi / p2-specific adrenergic receptor antagonist is propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing.

6. The method of claim 4, wherein the pi / p2-specific adrenergic receptor antagonist is propranolol or a pharmaceutically acceptable salt thereof.

7. The method of any one of claims 1 to 6, wherein the disease or disorder associated with elevated red blood cells is a myeloproliferative disorder.

8. The method of claim 7, wherein the myeloproliferative disorder is polycythemia vera (PV).

9. The method of claim 7 or claim 8, wherein the patient has a myeloproliferative disorder associated with a JAK2 mutation.

10. The method of claim 9, wherein the JAK2 mutation is a Jak2V617F mutation.

11. The method of any one of claims 7 to 10, wherein the myeloproliferative disorder has progressed to a cancer.

12. The method of any one of claims 1 to 6, wherein the disease or disorder associated with elevated red blood cells is secondary polycythemia.

13. The method of claim 12, wherein the secondary polycythemia is caused by sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastoma and meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

14. A method of decreasing red blood cells in a patient, the method comprising administering to the patient an effective amount of a p2-specific adrenergic receptor antagonist selected from ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

15. A method of decreasing erythroid differentiation in a patient, the method comprising administering to the patient an effective amount of a p2-specific adrenergic receptor antagonist selected from ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

16. A method of treating elevated red blood cells in a patient, the method comprising administering to the patient an effective amount of a p2-specific adrenergic receptor antagonist selected from ICI-118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

17. The method of any one of claims 14 to 16, wherein the patient has a disease or disorder associated with elevated red blood cells.

18. The method of claim 17, wherein the disease or disorder associated with elevated red blood cells is a myeloproliferative disorder.

19. The method of claim 18, wherein the myeloproliferative disorder is polycythemia vera (PV).

20. The method of claim 18 or claim 19, wherein the patient has a myeloproliferative disorder associated with a JAK2 mutation.

21. The method of claim 20, wherein the JAK2 mutation is a Jak2V617F mutation.

22. The method of any one of claims 18 to 21, wherein the myeloproliferative disorder has progressed to a cancer.

23. The method of claim 17, wherein the disease or disorder associated with elevated red blood cells is secondary polycythemia.

24. The method of claim 23, wherein the secondary polycythemia is caused by sleep apnea, smoking, lung disease, obesity, hypoventilation, Pickwickian syndrome, chronic obstructive pulmonary disease (COPD), diuretics, androgen intake, carbon monoxide poisoning, living at high altitude, kidney disease or cysts, a brain tumor selected from cerebellar hemangioblastoma and meningioma, a tumor of the parathyroid gland, hepatocellular cancer, renal cell cancer, an adrenal gland tumor, and / or benign uterine fibroids.

25. The method of any one of claims 1 to 24, wherein the method does not induce anemia in the patient.

26. The method of any one of claims 1 to 3 and 7 to 25, wherein the P2-specific adrenergic receptor antagonist is orally administered to the patient.

27. The method of any one of claims 4 to 25, wherein the pi / p2-specific adrenergic receptor antagonist is orally administered to the patient.

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

29. Use of 32-specific adrenergic receptor antagonist in the manufacture of a medicament for the treatment a disease or disorder associated with elevated red blood cells.

30. The use of claim 29, wherein the p2-specific adrenergic receptor antagonist is ICI- 118551, butaxamine, or a pharmaceutically acceptable salt of any of the foregoing.

31. Use of P l / p2-specific adrenergic receptor antagonist in the manufacture of a medicament for the treatment a disease or disorder associated with elevated red blood cells.

32. The use of claim 31, wherein the pi / p2-specific adrenergic receptor antagonist is propranolol, timolol, nadolol, sotalol, penbutolol, pindolol, or a pharmaceutically acceptable salt of any of the foregoing.

33. The use of claim 31, wherein the pi / p2-specific adrenergic receptor antagonist is propranolol or a pharmaceutically acceptable salt thereof.

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