Formulations of a hepcidin peptide analogue

Abstract

The present disclosure encompasses pharmaceutical compositions or formulations of hepcidin peptide analogues or mimetics, as well as to the use of the formulations in the treatment and / or prevention of a variety of diseases, conditions, or disorders, including the treatment and / or prevention of iron overload diseases including hereditary hemochromatosis, iron-loading anemias, and other conditions and disorders described herein.

Description

Attorney Docket No. T0947.70006WO00 FORMULATIONS OF A HEPCIDIN PEPTIDE ANALOGUE RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. § 119(e) of U.S. provisional application number U.S. provisional application number 63 / 570,697, filed March 27, 2024; U.S. provisional application number 63 / 709,550, filed October 21, 2024; U.S. provisional application number 63 / 764,237, filed February 27, 2025; and U.S. provisional application number 63 / 766,238, filed March 3, 2025, the contents of each of which are incorporated by reference herein in its entirety. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (T094770006WO00-SEQ-EMB.xml; Size: 11,173 bytes; and Date of Creation: March 25, 2025) are herein incorporated by reference in its entirety. FIELD

[0001] The present disclosure encompasses pharmaceutical compositions or formulations of hepcidin peptide analogues or mimetics, as well as the use of the formulations in the treatment and / or prevention of a variety of diseases, conditions, or disorders, including the treatment and / or prevention of iron overload diseases including hereditary hemochromatosis, iron-loading anemias such as thalassemia, diseases associated with ineffective or augmented erythropoiesis such as polycythemia vera, and other conditions and disorders described herein. BACKGROUND

[0002] Hepcidin (also referred to as LEAP-1), a peptide hormone produced by the liver, is a regulator of iron homeostasis in humans and other mammals. Hepcidin acts by binding to its receptor, the iron export channel ferroportin, causing its internalization and degradation. Human hepcidin is a 25-amino acid peptide (Hep25). See Krause et al. (2000) FEBS Lett 480:147-150, and Park et al. (2001) J Biol Chem 276:7806-7810. The structure of the bioactive 25-amino acid form of hepcidin is a simple hairpin with 8 cysteines that form 4 disulfide bonds as described by Jordan et al. J Biol Chem 284:24155-67. The N terminal region is required for iron-regulatory function, and deletion of 5 N-terminal amino acid residues results in a loss of iron-regulatory function. See Nemeth et al. (2006) Blood 107:328-33.Attorney Docket No. T0947.70006WO00

[0003] Abnormal hepcidin activity is associated with iron overload diseases, including hereditary hemochromatosis (HH) and iron-loading anemias. Hereditary hemochromatosis is a genetic iron overload disease that is mainly caused by hepcidin deficiency or in some cases by hepcidin resistance. This allows excessive absorption of iron from the diet and development of iron overload. Clinical manifestations of HH may include liver disease (e.g., hepatic cirrhosis and hepatocellular carcinoma), diabetes, and heart failure. Currently, the only treatment for HH is regular phlebotomy, which is very burdensome for the patients. Iron-loading anemias are hereditary anemias with ineffective erythropoiesis such as β-thalassemia, which are accompanied by severe iron overload. Complications from iron overload are the main cause of morbidity and mortality for these patients. Hepcidin deficiency is the main cause of iron overload in non-transfused patients and contributes to iron overload in transfused patients. The current treatment for iron overload in these patients is iron chelation, which is burdensome, sometimes ineffective, and accompanied by frequent side effects.

[0004] Polycythemia vera (PV) is a chronic, progressive trilineage clonal disorder signified by increased myeloid, erythroid, and megakaryocytic cell proliferation / accumulation and is characterized by the World Health Organization (WHO) as a myeloproliferative neoplasm (Arber et al., 2016, 127(20):2391-405). Diagnosis is defined by two criteria; the first being increased red blood cell mass, bone marrow biopsy showing trilineage hypercellularity and presence of JAK2V617F or JAK2 exon 12 mutations and the second criteria incorporates polycythemia, bone marrow biopsy confirmation and subnormal serum erythropoietin levels. Id. As PV is a disease characterized by increased erythropoiesis, it has been shown in animal models that high doses of hepcidin mimetics can ameliorate this disease by diminishing erythropoiesis (Casu et al., Blood.2016;128(2): 265-276).

[0005] Although therapeutic efficacy is the primary concern for a therapeutic agent, the pharmaceutical composition can be equally important to its development. Generally, drug developers endeavor to discover a pharmaceutical composition that possesses desirable properties, such as satisfactory water-solubility, storage stability, and reproducibility, all of which can impact the processability, manufacture, and / or bioavailability of the drug. Accordingly, discovery of pharmaceutical compositions that possess some or all of these desired properties is vital to drug development. The present disclosure provides pharmaceutical compositions or formulations that meet this and other needs. SUMMARY OF THE DISCLOSUREAttorney Docket No. T0947.70006WO00

[0006] Generally, the disclosure provides a pharmaceutical composition or a pharmaceutical formulation, which comprises a peptide having the amino acid sequence: Isovaleric acid- DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, wherein two cysteine residues of the peptide are linked via a disulfide bond, and wherein the pharmaceutical composition is a lyophilized solid form, a liquid form comprising reconstitution of the lyophilized solid form into solution, or a stable liquid form. The hepcidin mimetic peptide called rusfertide comprises the peptide sequence of SEQ ID NO: 1 and the two- dimensional chemical structure of Formula (I). Herein, rusfertide is also referred to as Compound 1 or PTG-300. See, for example, US Patent No.10,030,061, which is hereby incorporated-by- reference, including for its teachings on unnatural amino acid abbreviations and structures.

[0007] The inventors have surprisingly found that the addition of certain stabilizing agents, including zinc salts, enhances the pharmacokinetic and pharmacodynamic properties of liquid, lyophilized, and reconstituted lyophilized pharmaceutical formulations and in certain embodiments enhances the stability and reduces the presence of degradation products.

[0008] Accordingly, the disclosure provides a pharmaceutical composition or formulation which comprises a peptide having the amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu- Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, including a zinc salt, wherein the two cysteine residues of the peptide are cyclized to form a disulfide bond.

[0009] In another aspect, the disclosure provides a method for preparing a lyophilized pharmaceutical composition, the method comprising: preparing a solution comprising a peptide having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, a bulking agent, a buffering agent and optionally a zinc salt at a suitable pH, wherein the two cysteine residues of the peptide are cyclized to form a disulfide bond; and drying the solution by lyophilization to afford a lyophilized pharmaceutical composition.

[0010] In another aspect, the disclosure provides a method for preparing a pharmaceutical composition or formulation suitable for subcutaneous administration, the method comprising mixing or reconstituting a lyophilized pharmaceutical composition with a diluent to obtain a solution suitable for subcutaneous administration, wherein the lyophilized pharmaceutical composition comprises a peptide having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Attorney Docket No. T0947.70006WO00 Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt, and optionally a first zinc salt; the diluent comprises optionally a second zinc salt and a solvent; and wherein the two cysteine residues of the peptide are linked to form a disulfide bond.

[0011] In another aspect, the disclosure provides a method for treating a disease of iron metabolism in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein.

[0012] In some embodiments, the disclosure provides a method for treating polycythemia vera in a subject in need thereof, the method comprising subcutaneously administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein.

[0013] In various embodiments, the disclosure generally encompasses a pharmaceutical formulation comprising: (i) a peptide comprising an amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 SEQ ID NO: 1 or a pharmaceutically acceptable salt or solvate thereof, and (ii) a zinc salt, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:1.

[0014] In certain embodiments, the peptide comprises a disulfide bond between two cysteine residues of the peptide.

[0015] In certain embodiments, the peptide comprises a structure of formula (I):Attorney Docket No. T0947.70006WO00 or a pharmaceutically acceptable salt or hydrate thereof.

[0016] A peptide comprising the structure of Formula (I) and the sequence of SEQ ID NO: 1 is the peptide named rusfertide, which is in clinical trials for the treatment of polycythemia vera. See, e.g., Kremyanskaya, M., et al., Rusfertide, a Hepcidin Mimetic, for Control of Erythrocytosis in Polycythemia Vera, N Engl J Med 2024; 390:723-735; DOI: 10.1056 / NEJMoa230880. Thus, any formulation disclosed herein may comprise rusfertide.

[0017] In certain embodiments, the molar ratio of the zinc salt and the peptide is from about 0.2:1 to about 2.5:1.

[0018] In certain embodiments, the molar ratio of the zinc salt and the peptide is from about 0.3:1 to about 1.2:1.

[0019] In certain embodiments, the molar ratio of the zinc salt and the peptide is from about 0.75:1 to about 1:1.

[0020] In certain embodiments, the zinc salt is zinc maleate, zinc tartrate, zinc oxide, zinc hydroxide, zinc mesylate, zinc besylate, zinc chloride or zinc acetate.

[0021] In certain embodiments, the zinc salt is zinc acetate.

[0022] In certain embodiments, the zinc salt is zinc chloride.

[0023] In certain embodiments, the composition is a stable liquid formulation. As described herein, a “stable” formulation (whether in liquid or lyophilized form) generally refers to its percent or degree of purity with respect to the active ingredient, i.e., the peptide, over time. Especially at room temperature or at higher temperatures, peptides can degrade, for example, into smaller peptide forms. Thus, a “stable liquid formulation” of the disclosure is a liquid solution comprising the peptide of SEQ ID NO: 1 and / or having the structure of Formula (I), with a specified level of stability or purity. For example, in some embodiments, a “stable liquid formulation” of the disclosure comprises said peptide and has a peptide purity of at least 95%, 96%, 97%, 98%, or 99% for at least 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 24 months at 4 ℃, 5 ℃, 25 ℃ or at ambient room temperature.

[0024] In certain embodiments, the composition is a lyophilized formulation. In certain embodiments, the composition is a reconstituted lyophilized formulation. A reconstituted lyophilized formulation is a type of liquid formulation.

[0025] In certain embodiments, a reconstituted lyophilized formulation comprising a zinc salt improves bioavailability as compared to a liquid formulation that lacks the zinc salt.Attorney Docket No. T0947.70006WO00

[0026] In certain embodiments, a stable liquid formulation improves bioavailability and / or stability of the peptide as compared to liquid formulations without a zinc salt or even as compared to the lyophilized / reconstituted-lyophilized formulations of the disclosure. This provides the opportunity to reduce therapeutic doses. In certain embodiments, a stable liquid formulation allows the formulation to be included in an autoinjector as the improved stability of the stable liquid formulation allows for longer term storage without having to rely on reconstitution of a lyophilized cake. This can improve patient convenience and compliance, dosing accuracy, and treatment adherence as the patient (or doctor) does not need to reconstitute a lyophilized cake with a solution for filling a syringe. However, this does not therefore mean that the lyophilized and reconstituted lyophilized formulations of the disclosure are not superior to prior peptide formulations. For example, as shown herein, reconstituted lyophilized formulations comprising a zinc salt and the peptide of SEQ ID NO: 1 or having the structure of Formula (I) exhibits improved bioavailability for said peptide over a liquid formulation (“PFS”; pre-filled syringe with said peptide) that did not contain a zinc salt. And because the peptide is in lyophilized form until reconstitution, sufficient stability / purity of the peptide can be maintained. Further, in certain embodiments, the lyophilized formulation, reconstituted lyophilized formulation, or the stable liquid formulation allows lower doses that potentially reduce side effects.

[0027] In certain embodiments, the composition maintains a purity of greater than 97% at room temperature for at least 12 months.

[0028] In certain embodiments, the composition maintains a purity of greater than 95% at room temperature for at least 24 months.

[0029] In certain embodiments, the pharmaceutical composition comprises a tonicity adjusting agent, a buffering agent, and a stabilizer.

[0030] In certain embodiments, the pharmaceutical composition further comprises a buffering agent and a bulking agent.

[0031] In certain embodiments, the composition is a solution comprising one or more solvents.

[0032] In certain embodiments, the solvent includes water, dimethylsulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide, or a combination thereof.

[0033] In certain embodiments, the peptide in the composition is present in an amount from about 0.1% w / v to about 15% w / v.Attorney Docket No. T0947.70006WO00

[0034] In certain embodiments, the peptide in the composition is from about 1% w / v to about 6% w / v.

[0035] In certain embodiments, the peptide in the composition is from about 2% w / w to about 12% w / w.

[0036] In certain embodiments, the dosage of the peptide is from about 1 mg to about 100 mg.

[0037] In certain embodiments, the dosage of the peptide is about 1 mg, about 5 mg, about 10 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg.

[0038] In other embodiments, the disclosure encompasses a liquid pharmaceutical composition comprising a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu- Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, a buffering agent, a zinc salt, a stabilizer, a tonicity adjuster, a pH adjuster, and a solvent, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:1.

[0039] In certain embodiments, the buffering agent comprises sodium acetate, the zinc salt comprises zinc chloride, the stabilizer comprises PEG 3350, the tonicity adjuster comprises sorbitol, the pH adjuster comprises sodium hydroxide and acetic acid, and the solvent comprises water.

[0040] In certain embodiments, the liquid composition has purity greater than 97% after 12 months.

[0041] In certain embodiments, the li quid composition has less than about 2.0 % Asp-related degradants after 24 months.

[0042] In certain embodiments, the liquid composition has a purity greater than 97% after 12 months and less than about 2.0% Asp-related degradants after 24 months.

[0043] In certain embodiments, the liquid pharmaceutical composition comprises: Table a – Liquid Pharmaceutical Composition 1 Component Amount (w / v)Attorney Docket No. T0947.70006WO00 (Zn:PTG-300 molar (0.75:1 to 3:1)

[0044] In certain embodimentprises: Table b – Liquid Pharmaceutical Composition 2 Component Amount (w / v)

[0045] In certain embodiments, e qu p armaceu ca compos on comprses: Table c – Liquid Pharmaceutical Composition 3 Component Amount (w / v)Attorney Docket No. T0947.70006WO00 Sodium Acetate Trihydrate 0.272%

[0046] In another embodimentpharmaceutical composition comprising a peptide having an amino acid sequence of Isovaleric acid- DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, a bulking agent, and a zinc salt, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:1.

[0047] In certain embodiments, the bulking agent is mannitol, and wherein the zinc salt comprises zinc acetate.

[0048] In certain embodiments, the composition is reconstituted for injection and comprises: Table d – Liquid Pharmaceutical Composition - 4 Component Amount (w / v)

[0049] In certain embodiments, the dosage of the peptide is about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg.

[0050] The disclosure also encompasses a kit for subcutaneous injection comprising a lyophilized pharmaceutical composition comprising a peptide having an amino acid sequence of Isovaleric acid- DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptableAttorney Docket No. T0947.70006WO00 salt or solvate thereof, and a reconstitution liquid comprising a zinc salt, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:1. In some embodiments, both the lyophilized composition and the reconstitution liquid comprises a zinc salt, where their combined zinc amount is present in a molar ratio to the peptide at about 0.1:1 to about 3:1. In some embodiments, only the reconstitution liquid comprises a zinc salt.

[0051] In certain embodiments, the kit will include a lyophilized powder for reconstitution with a provided diluent. Both the peptide and diluent are filled in 2 mL glass vials with a butyl rubber septum held in place with an aluminum overseal. In certain embodiments, the vials are provided with vial adapters to facilitate transfer of the diluent and subsequent drug solution. In certain embodiments, a syringe and a needle are supplied and packaged with the vials in a carton as a kit for reconstitution. In certain embodiments, the lyophilized peptide is stored at 2°C to 8°C, in an opaque kit carton which affords protection from light.

[0052] In certain embodiments, the liquid formulations include a 2:1 zinc chloride : peptide molar ratio for subcutaneous administration that can be prepared by a pharmacist at the clinical site according to instructions provided. In certain embodiments, the peptide is stored at -20°C (-15°C to -25°C), protected from light. In certain embodiments, the dose volume is 0.5 ml for administration.

[0053] In certain embodiments, the liquid formulations include a 2:1 zinc chloride : peptide molar ratio for subcutaneous administration, wherein the liquid formulations are already prepared in an autoinjector, thereby alleviating the need for any dose preparation by the pharmacist, doctor, or patient. In certain embodiments, the dose volume is 0.5 ml for administration.

[0054] In another embodiment, the disclosure encompasses a method for treating a disease of iron metabolism in a subject, the method comprising administering to the subject a therapeutically effective amount of a liquid pharmaceutical composition of any one of claims 1-35.

[0055] In certain embodiments, the disease of iron metabolism is an iron overload disease.

[0056] In other embodiments, the disclosure encompasses a method for treating polycythemia vera in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a pharmaceutical composition.

[0057] In certain embodiments, the formulation is administered by subcutaneous injection.

[0058] In certain embodiments, the subcutaneous injection is administered using an autoinjector.

[0059] In certain embodiments, the composition is administered with a dosing regimen to achieve: an average peptide plasma area under the curve (AUC) of at least 32000 h*ng / mL for a compositionAttorney Docket No. T0947.70006WO00 comprising about 20 mg, 30 mg, 45 mg or 60 mg of peptide; and an average maximum peptide blood plasma concentration (average Cmax) of at least 500 ng / mL per each dosage of 20 mg, 30 mg, 45 mg or 60 mg of peptide delivered; wherein the AUC is measured from time zero to the time of last measured concentration.

[0060] In certain embodiments, the zinc salt is zinc chloride.

[0061] In certain embodiments, the zinc salt and peptide in the composition have a molar ratio of 0.2:1, 1:1, or 2:1.

[0062] In certain embodiments, the peptide is administered once weekly or twice weekly.

[0063] In certain embodiments, the peptide in the composition is present in an amount from about 0.1% w / v to about 15% w / v.

[0064] In certain embodiments, the peptide in the composition is from about 1% w / v to about 6% w / v.

[0065] In certain embodiments, the peptide in the composition is from about 2% w / w to about 12% w / w.

[0066] In certain embodiments, the dosage of the peptide is from about 1 mg to about 100 mg.

[0067] In certain embodiments, the dosage of the peptide is about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg or about 100 mg. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] Figure 1A illustrates changes of mean plasma concentration of Compound 1 over time in patients administered with various reconstituted lyophilized doses of the compound (10, 20, 30, 45, and 60 mg doses; see Tables 1-3 and Example 4 for formulation recipes; these “Lyo” formulations were used in Figures 1-6).

[0069] Figure 1B illustrates changes of median plasma concentration of Compound 1 over time in patients administered with various doses of the compound.

[0070] Figure 2A shows the exposure of reconstituted lyophilized Compound 1 is dose related.

[0071] In Figure 2B, the exposures of Compound 1 in individual subjects are presented, where each subject was administered with different doses.

[0072] Figure 3A illustrates the comparisons of the mean plasma concentrations of reconstituted lyophilized Compound 1 for patients in groups 1 and 2. They were administered with PFSAttorney Docket No. T0947.70006WO00 formulations containing 20 mg of Compound 1 versus reconstituted lyophilized formulations also containing 20 mg of Compound 1.

[0073] Figure 3B shows the comparisons of median plasma concentrations of Compound 1 for patients in groups 1 and 2, who were administered with the same PFS formulations and reconstituted lyophilized formulations, both containing 20 mg of Compound 1.

[0074] Figure 4A illustrates the comparisons of mean plasma iron concentrations in patients from groups 1 and 2. These patients were administered with PFS formulation versus reconstituted lyophilized formulations containing different doses of Compound 1, namely 10 mg, 20 mg, 30 mg, 45 mg or 60 mg.

[0075] Figure 4B shows the comparisons of the median plasma iron concentrations in patients from groups 1 and 2. These patients were administered with PFS formulations versus reconstituted lyophilized formulations containing 10 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1.

[0076] Figures 5A-5D illustrate the purities of reconstituted lyophilized formulations and PFS formulations. Figure 5A shows the purity profiles of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg Compound 1 at 5 ºC over approximately 24 months.

[0077] Figure 5B presents a comparison of the purities of a PFS liquid formulation containing 30 mg Compound 1 and a reconstituted lyophilized formulation also containing 30 mg of Compound 1 at 5 ºC over approximately 24 months.

[0078] Figure 5C displays the purity profiles of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg or 60 mg Compound 1 at 25 ºC over approximately 24 months.

[0079] Figure 5D provides a comparison of the purities of a PFS liquid formulation containing 30 mg Compound 1 and reconstituted lyophilized formulation also containing 30 mg of Compound 1. The comparison is based on purity at 25 ºC over a 24-month time period.

[0080] Figure 6A shows the Asp-related data of a reconstituted lyophilized formulations containing various doses of Compound 1 (10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg) at 5 ºC over a period of approximately 24 months.

[0081] Figure 6B presents a comparison of the asp-related degradant of a PFS liquid formulation containing 30 mg Compound 1 and a reconstituted lyophilized formulation also containing 30 mg of Compound 1 at 5 ºC over approximately 24 months.Attorney Docket No. T0947.70006WO00

[0082] Figure 6C displays the asp-related degradant of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg or 60 mg Compound 1 at 25 ºC over approximately 24 months.

[0083] Figure 6D provides a comparison of the purities of a PFS liquid formulation containing 30 mg Compound 1 and reconstituted lyophilized formulation also containing 30 mg of Compound 1. The comparison is based on asp-related degradant at 25 ºC over a 24-month time period.

[0084] Figure 7 illustrates the changes in mean plasma concentration of Compound 1 over time in subjects who were administered a 20 mg dose of Compound 1. The subjects received Compound 1 in one lyophilized formulation and three stable liquid formulations with different molar ratios of zinc and Compound 1.

[0085] Figure 8 illustrates the changes in mean plasma concentration of Compound 1 over time in subjects who were administered a 45 mg dose of Compound 1. The subjects received Compound 1 in a lyophilized formulation and three stable liquid formulations with different molar ratios of zinc and Compound 1.

[0086] Figure 9 shows a comparison of the changes in mean plasma concentration of Compound 1 over time in subjects who were administered a 20 mg dose of Compound 1. The subjects received Compound 1 in a PFS formulation and a stable liquid formulation containing ZnCl2 with a 2:1 molar ratio of zinc and Compound 1.

[0087] Figure 10A illustrates the stabilities of four stable liquid formulations and a PFS formulation at 5 ºC over a 9-month period. The stability was assessed by monitoring the purity of the formulations over time. As depicted in the graphs, the stable liquid formulations contain 10 mg / mL, 30 mg / mL, 60 mg / mL, and 120 mg / mL of Compound 1, respectively, with a molar ratio of zinc and Compound 1 of 2:1.

[0088] Figure 10B presents the stabilities of four stable liquid formulations and a PFS formulation at 5 ºC over a 9-month period. The stability was assessed based on the Asp-related degradants observed over time. As indicated in the graphs, the stable liquid formulations contain 10 mg / mL, 30 mg / mL, 60 mg / mL, and 120 mg / mL of Compound 1, respectively, with a molar ratio of zinc and Compound 1 of 2:1.

[0089] Figure 10C illustrates the stabilities of four stable liquid formulations and a PFS formulation at 5 ºC over a 9-month period. The stability was assessed based on the total degradants observed over time. As depicted in the graphs, the stable liquid formulations contain 10 mg / mL, 30 mg / mL,Attorney Docket No. T0947.70006WO00 60 mg / mL, and 120 mg / mL of compound 1, respectively, with a molar ratio of zinc and Compound 1 of 2:1.

[0090] Figure 11A illustrates the stabilities of three stable liquid formulations with different zinc and Compound 1 ratios and a PFS formulation at 5 ºC over a 9-month period. The stability was determined based on the purity of the formulations over time. As shown in the graphs, the stable liquid formulations contain 30 mg / mL of Compound 1, with molar ratios of zinc to Compound 1 of 2:1, 1:1 or 0.

[0091] Figure 11B illustrates the stabilities of three stable liquid formulations with different zinc and Compound 1 ratios and a PFS formulation at 5 ºC over a 9-month period. The stability was assessed based on the Asp-related degradants observed over time. As shown in the graphs, the stable liquid formulations contain 30 mg / mL of Compound 1, with a molar ratio of zinc and Compound 1 of 2:1, 1:1 or 0.

[0092] Figure 11C illustrates the stabilities of three stable liquid formulations with different zinc and Compound 1 ratios and a PFS formulation at 5 ºC over a 9-month period. The stability was assessed based on the total degradants observed over time. As shown in the graphs, the stable liquid formulations contain 30 mg / mL of Compound 1, with a molar ratio of zinc and Compound 1 of 2:1, 1:1 or 0.

[0093] Figure 12A illustrates the concentration of rusfertide in nanograms per milliliter (ng / mL) over time in hours (h) following a single subcutaneous (SC) dose of rusfertide in either aqueous or lyophilized formulations. The formulation is lyophilized if it is not indicated as aqueous.

[0094] Figure 12B illustrates the concentration of rusfertide metabolite 4 (M4) following the dosing described in Figure 12A.

[0095] Figure 12C illustrates the concentration of rusfertide metabolite 9 (M9) following the dosing described in Figure 12A.

[0096] Figure 13A illustrates the change in serum iron levels in micromoles per liter (µmol / L) following the dosing described in Figure 12A.

[0097] Figure 13B illustrates the change in percentage (%) transferrin-iron saturation (TSAT) following the dosing described in Figure 12A.

[0098] Figure 14A illustrates rusfertide maximum concentration (Cmax) following a single-dose administration of rusfertide. “mg” is milligrams.Attorney Docket No. T0947.70006WO00

[0099] Figure 14B illustrates the area under the curve extrapolated to infinite time (AUCinf) following a single-dose administration of rusfertide. “ng·h / mL” is nanograms per hour per milliliter.

[0100] Figure 15A illustrates the serum iron concentration as a result of rusfertide administration. Rusfertide concentration was measured as area under the curve from 0-168 hours. AUECtis the area under the effect curve over time and reflects the change in serum iron concentration over time after rusfertide administration.

[0101] Figure 15B illustrates transferrin iron saturation (TSAT) as a result of rusfertide administration. Rusfertide concentration was measured as area under the curve from 0-168 hours. AUECt is the area under the effect curve over time and reflects the change in TSAT over time after rusfertide administration.

[0102] Figure 16A illustrates the change in serum iron concentration as a function of rusfertide concentration.

[0103] Figure 16B illustrates the change in transferrin iron saturation (TSAT) as a function of rusfertide concentration. DETAILED DESCRIPTION

[0104] In general, the present disclosure relates to hepcidin peptide analogues or mimetics or pharmaceutically acceptable salt or solvate forms thereof, corresponding pharmaceutical compositions, methods and / or uses for treatment of iron metabolism related diseases and disorders. Definitions

[0105] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0106] Throughout the present specification, the terms “about” and / or “approximately” may be used in conjunction with numerical values and / or ranges. The term “about” is understood to mean those values near to a recited value. For example, “about 40 [units]” may mean within ± 25% of 40 (e.g., from 30 to 50), within ± 20%, ± 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1%, less than ± 1%, or any other value or range of values therein or there below. Furthermore, the phrases “less than about [a value]” or “greater than about [a value]” should be understood in view of the definition of the term “about” provided herein. The terms “about” and “approximately” may beAttorney Docket No. T0947.70006WO00 used interchangeably. In some embodiments, “About” when referring to a value includes the stated value + / - 10% of the stated value. For example, about 50% includes a range of from 45% to 55%, while about 20 molar equivalents includes a range of from 18 to 22 molar equivalents. As such, about 20 molar equivalents includes values of 18, 19, 20, 21 and 22 molar equivalents.

[0107] “Administering” refers to administration of the composition of the present disclosure to a subject.

[0108] “Composition” as used herein is intended to encompass a product that includes the specified active ingredient and pharmaceutically acceptable excipients, carriers or diluents as described herein, such as in specified amounts defined throughout the originally filed disclosure, which results from combination of specific components, such as specified ingredients in the specified amounts as described herein.

[0109] Throughout the present specification, numerical ranges are provided for certain quantities. It is to be understood that these ranges comprise all subranges therein. Thus, the range “from 50 to 80” includes all possible ranges therein (e.g., 51-79, 52-78, 53-77, 54-76, 55-75, 60-70, etc.). Furthermore, all values within a given range may be an endpoint for the range encompassed thereby (e.g., the range 50-80 includes the ranges with endpoints such as 55-80, 50-75, etc.).

[0110] It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely", "only" and the like in connection with the recitation of claim elements, or the use of a "negative" limitation.

[0111] The term “a” or “an” refers to one or more of that entity; for example, “a kinase inhibitor” refers to one or more kinase inhibitors or at least one kinase inhibitor. As such, the terms “a” (or “an”), “one or more” and “at least one” are used interchangeably herein. In addition, reference to “an inhibitor” by the indefinite article “a” or “an” does not exclude the possibility that more than one of the inhibitors is present, unless the context clearly requires that there is one and only one of the inhibitors.

[0112] As used herein, the verb “comprise” as is used in this description and in the claims and its conjugations are used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. The present disclosed compositions, formulations, methods, and uses may suitably “comprise”, “consist of”, or “consist essentially of”, the steps, elements, and / or reagents described in the claims.Attorney Docket No. T0947.70006WO00

[0113] As used herein, the terms “Compound 1,” “Compound of Formula I,” are used interchangeably and refer to a compound including the sequence: Isovaleric acid- DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, or having the structural formula:inhibits the normal cellular activities of the proteins that are involved, that constitute, or that are participants in the cellular pathways.

[0115] As used herein, the term "subject," "individual" or "patient" is used interchangeably and refers to a vertebrate, preferably a mammal. Non-limiting examples include mice, dogs, rabbits, farm animals, sport animals, pets, and humans.

[0116] The term "treat," ''treated," "treating" or ''treatment" includes the diminishment or alleviation of at least one symptom associated or caused by the state, disorder or disease being treated. Treatment can be diminishment of one or several symptoms of a disorder or complete eradication of a disorder or a disease.

[0117] As used herein, ''therapeutically effective amount" or an “effective amount” indicates an amount that results in a desired pharmacological and / or physiological effect for the condition. The effect may be prophylactic in terms of completely or partially preventing a condition or symptom thereof and / or may be therapeutic in terms of a partial or complete cure for the condition and / or adverse effect attributable to the condition. In some embodiments, a therapeutically effective dose ofAttorney Docket No. T0947.70006WO00 a peptide for polycythemia vera is an amount sufficient to reduce or maintain the hematocrit of the polycythemia vera subject to less than about 45% until the next the dose.

[0118] As used herein, the terms “synergy” or “synergistic effect” means that the therapeutic effect of the compounds, therapeutics, or drugs when used in combination is greater than the additive therapeutic effects of the compounds when used individually. In one example, a synergy between compounds V and W is observed when the combination of the two compounds reduces cell proliferation by 80% whereas compound V alone only reduces cell proliferation by 20% and compound W alone only reduces cell proliferation by 15%. The 80% reduction in cell proliferation exceeds the expected additive effect of 35%; thus, compounds V and W show synergistic effect for reducing cell proliferation.

[0119] By “pharmaceutically acceptable” it is meant the carrier(s), diluent(s) or excipient(s) must be compatible with the other components or ingredients of the compositions of the present disclosure, i.e., that which is useful, safe, non-toxic acceptable for pharmaceutical use. In accordance with the present disclosure, pharmaceutically acceptable means approved or approvable as is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans.

[0120] As used herein, the term “pharmaceutically acceptable salt” refers to salts which are known to be non-toxic and are commonly used in the art of pharmaceutical formulation. Typical inorganic acids used to form such salts include, but are not limited to, hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, phosphoric and hypophosphoric acid and the like. Salts derived from organic acids include, but are not limited to, aliphatic mono and dicarboxylic acids, phenylsubstituted alkanoic acids, hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphatic and aromatic sulfonic acids. Examples of pharmaceutically acceptable salts thus include, but are not limited to, acetate, phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate, beta-hydroxybutyrate, chloride, cinnamate, citrate, formate, fumarate, glycolate, heptanoate, lactate, maleate, hydroxymaleate, malonate, mesylate, nitrate, oxalate, phthalate, phosphate, monohydro genphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, propionate, phenylpropionate, salicylate, succinate, sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate, benzenesulfonate, p-bromophenylsulfonate, chlorobenzenesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate, naphthalene-1-sulfonate,Attorney Docket No. T0947.70006WO00 naphthalene-2-sulfonate, p-toluenesulfonate, xylenesulfonate, tartarate, and the like. In certain embodiments, a pharmaceutically acceptable salt of the peptide of formula (I) is e.g., an acetate salt.

[0121] A pharmaceutically acceptable carrier, diluent or excipient refers to a non-toxic solid, semi- solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride, and the like.

[0122] The term “pharmaceutically acceptable carrier” includes any of the standard pharmaceutical carriers. Pharmaceutically acceptable carriers for therapeutic use are well known in the pharmaceutical art and are described, for example, in “Remington's Pharmaceutical Sciences”, 17th edition, Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, PA, USA, 1985. For example, sterile saline and phosphate-buffered saline at slightly acidic or physiological pH may be used. Suitable pH-buffering agents may, e.g., be phosphate, citrate, acetate, tris(hydroxymethyl)aminomethane (TRIS), N-tris(hydroxymethyl)methyl-3-aminopropane-sulfonic acid (TAPS), ammonium bicarbonate, diethanolamine, histidine, arginine, lysine or acetate (e.g. as sodium acetate), or mixtures thereof. The term further encompasses any carrier agents listed in the US Pharmacopeia for use in animals, including humans.

[0123] As used herein, the term "solvate" or “pharmaceutically acceptable solvate” refers to a complex formed by combination of solvent molecules with molecules or ions of the solute. The solvent can be an organic compound, an inorganic compound, or a mixture of both. Solvate is meant to include hydrate. "Hydrate" refers to a complex formed by combination of water molecules with molecules or ions of the solute. Suitable solvents for forming a solvate include, but are not limited to, protic solvents, polar aprotic solvents, and non-polar. For example, types of solvent may include aromatic hydrocarbons (e.g. toluene), chlorinated hydrocarbons, ethers (e.g. tetrahydrofuran), aliphatic hydrocarbons, alcohols (e.g. isopropanol), esters (e.g. ethyl acetate), ketones (e.g. acetone or methylethylketone), amides (e.g. dimethylformamide), nitriles (e.g. acetonitrile), sulfur- containing compounds (e.g. dimethylsulfoxide) and water, as well as cyclic and non-cyclic, aromatic and non-aromatic amines (e.g. pyridine or triethylamine). Some examples of solvents include, but are not limited to, methanol, N,N-dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and water. In general, the solvated forms are equivalent to unsolvated forms and are encompassed within the scope of the present disclosure. Certain compounds of the present disclosure may exist inAttorney Docket No. T0947.70006WO00 multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present disclosure and are intended to be within the scope of the present disclosure. In some embodiments, a solvate is a crystal form that contains either stoichiometric or non-stoichiometric amounts of solvent associated with the pharmaceutical ingredient or a compound it is referring to.

[0124] The term “L-amino acid,” as used herein, refers to the “L” isomeric form of a zinc salt, and conversely the term “D-amino acid” refers to the “D” isomeric form of a zinc salt. The zinc salt residues described herein are preferred to be in the “L” isomeric form, however, residues in the “D” isomeric form can be substituted for any L-amino acid residue, as long as the desired functionality is retained by the peptide.

[0125] The term “cyclized,” as used herein, refers to a reaction in which one part of a polypeptide molecule becomes linked to another part of the polypeptide molecule (or a different polypeptide molecule) to form a closed ring, such as by forming a disulfide bridge or other similar bond, e.g. a lactam bond. In particular embodiments, peptide monomer compounds or monomer subunits of peptide dimer compounds described herein are cyclized via an intramolecular bond between two amino acid residues present in the peptide monomer or monomer subunit.

[0126] “Thiol”, “mercapto” or “sulfanyl” means an -SH group.

[0127] The term “disulfide bond” means a -S-S- linkage.

[0128] The term “linker moiety” or “linker” as used herein, refers broadly to an amino acid, a peptide, or a doubly functionalized chemical structure that is capable of linking or joining together two chemical groups. In some embodiments, linking or link means two amino acid residues in the same peptide are covalently joined together through a linker moiety or linker to form a cyclic structure.

[0129] As used herein, “lyophilization” means freeze drying, which is a process where water is removed from a product after it is frozen and placed under a vacuum, allowing the ice to change directly from solid to vapor without passing through a liquid phase. In some embodiments, the process consists of freezing, primary drying (sublimation), and secondary drying (desorption).

[0130] The term “co-administration” or “coadministration” refers to administration of a peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and (b) at least one combination partner, together in a coordinated fashion. For example, the co-administration can be simultaneous administration, sequential administration, overlappingAttorney Docket No. T0947.70006WO00 administration, interval administration, continuous administration, or a combination thereof. In one embodiment, the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and at least one combination partner, also for treatment of integrin-related disorders, are formulated into a single dosage form. In another embodiment, the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and at least one combination partner are provided in a separate dosage forms.

[0131] By “simultaneous administration”, it is meant that the peptide, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and at least one combination partner are administered on the same day. For simultaneous administration, the peptide, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and at least one combination partner can be administered at the same time or one at a time.

[0132] The term “peptide,” as used herein, refers broadly to a sequence of two or more amino acids joined together by peptide bonds. It should be understood that this term does not connote a specific length of a polymer of amino acids, nor is it intended to imply or distinguish whether the polypeptide is produced using recombinant techniques, chemical or enzymatic synthesis, or is naturally occurring.

[0133] For the most part, the names of naturally occurring and non-naturally occurring aminoacyl residues used herein follow the naming conventions suggested by the IUPAC Commission on the Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on Biochemical Nomenclature as set out in “Nomenclature of α-Amino Acids (Recommendations, 1974)” Biochemistry, 14(2), (1975). To the extent that the names and abbreviations of amino acids and aminoacyl residues employed in this specification and appended claims differ from those suggestions, they will be made clear to the reader. Some abbreviations useful in describing the compositions and formulations are defined below in the following Table A. TABLE A. Definitions and Abbreviations Abb i ti D fi itiAttorney Docket No. T0947.70006WO00

[0134] As used herein, purity means the quality or state of being pure of a substance free from impurities. Purity can be determined by UHPLC with UV detection, and it is determined as the ratioAttorney Docket No. T0947.70006WO00 of the main peptide of formula (I) peak area relative to a sum of all the peak areas (including peptide of formula (I) and all its UV absorbing impurities and degradants).

[0135] As used herein, the term “Asp-related” degradants are a subset of the peaks measured in the purity by UHPLC analytical procedure, which include Iso-Asp (the primary degradant of the peptide of formula (I) and the aspartimide intermediate from the chemical degradation process of conversion from peptide of formula (I) to Iso-Asp.

[0136] As used herein, the term “solid form” means a physical form which is not predominantly in a liquid or a gaseous state. Solid forms may be crystalline, amorphous or mixtures thereof. As used herein and unless otherwise specified, the term “crystal forms” and related terms refer to solid forms that are crystalline. Crystal forms include, but are not limited to, non-solvates, non-hydrates, solvates, hydrates, and other molecular complexes, as well as salts, solvates of salts, hydrates of salts, and other molecular complexes of salts thereof. In certain embodiments, a solid form or crystal form of a substance may be substantially free of amorphous forms and / or other solid forms and / or crystal forms. In certain embodiments, a solid form and / or crystal form of a substance may contain less than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% on a weight basis of one or more amorphous forms and / or other solid forms and / or crystal forms on a weight basis. In certain embodiments, a solid form or crystal form of a substance may be physically and / or chemically pure. In certain embodiments, a solid form or crystal form of a substance may be about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and / or chemically pure on a weight basis. In certain embodiments, a solid form or crystal form of a substance may be at least about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and / or chemically pure on a weight basis. In one embodiment, a solid form or crystal form of a substance is about 97.0% physically and / or chemically pure on a weight basis. In one embodiment, a solid form or crystal form of a substance is about 98.0% physically and / or chemically pure on a weight basis. In one embodiment, a solid form or crystal form of a substance is about 99.0% physically and / or chemically pure on a weight basis. In one embodiment, a solid form or crystal form of a substance is about 99.9% physically and / or chemically pure on a weight basis. In certain embodiments, a solid form or crystal form may be substantially chemically pure and / or substantially physically pure on a weight basis.

[0137] As used herein “amorphous” means a solid form of a molecule, atom, and / or ions that is not crystalline. In particular, the term “amorphous form” describes a disordered solid form, i.e., a solidAttorney Docket No. T0947.70006WO00 form lacking long range crystalline order. An amorphous solid does not display a definitive X-ray diffraction pattern. In certain embodiments, an amorphous form of a substance may be substantially pure of other amorphous forms and / or crystal forms.

[0138] The abbreviation, “(w / w)” refers to the phrase “weight for weight” or “weight by weight”, i.e., the proportion of a particular substance within a mixture, as measured by weight or mass or a weight amount of a component of the composition disclosed herein relative to the total weight amount of the composition. Accordingly, the quantity is unit less or unitless and represents a weight percentage amount of a component relative to the total weight of the composition. For example, a 2% (w / w) solution means 2 grams of solute is dissolved in 100 grams of solution.

[0139] Bioavailability refers to the extent and rate at which the active moiety (drug or metabolite) enters systemic circulation, thereby accessing the site of action. Bioavailability of a drug is impacted by the properties of the dosage form, which depend partly on its design and manufacture.

[0140] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the disclosed compositions, formulations, methods, and uses belong. Preferred methods, devices, and materials are described, although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosed compositions, formulations, methods, and uses. Pharmaceutical Compositions and Methods

[0141] In one aspect, the present disclosure provides a pharmaceutical composition or formulation comprising a peptide, i.e., a compound of formula (I), or a peptide of formula (I), or Compound 1 or a pharmaceutically acceptable salt or solvate thereof, and a bulking agent and a buffering agent. As used herein, the terms “Compound of formula (I)”, “peptide of formula (I)” and “Compound 1” are used interchangeably and refer to the same peptide molecule. The peptide comprises or consists of an amino acid sequence defined as: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1), wherein the two cysteine residues of the peptide are linked via a disulfide bond, for example, the thio or mercapto group on the side chains of the two cysteine residues in the peptide are taken together to form a disulfide bond, i.e., a -S-S- bond. In another word, the peptide is cyclized through a disulfide bond formed by the two cysteine residues in the peptide. The chemical structure of the compound of formula (I) is shown below:Attorney Docket No. T0947.70006WO00 salt or solvate thereof can behydrate, or other solvate. The peptide of formula (I) or SEQ ID NO: 1 or a pharmaceutically acceptable salt or solvate form thereof can be in a crystalline form, an amorphous form, or a semi-crystalline form.

[0143] In some embodiments, the disclosure provides a pharmaceutical composition or formulation comprising a peptide of formula (I) or a pharmaceutically acceptable salt or solvate thereof, and a zinc salt.

[0144] In some embodiments, the disclosure provides a pharmaceutical composition or formulation, which is in a lyophilized solid form. The solid form can be amorphous, crystalline, or semi- crystalline. In one embodiment, the pharmaceutical composition is a lyophilized powder. In one embodiment, the solid form is amorphous.

[0145] In some embodiments, the disclosure provides a pharmaceutical composition comprising a peptide of formula (I) or a pharmaceutically acceptable salt or solvate thereof, and a bulking agent and a buffering agent, wherein the pharmaceutical composition is a lyophilized solid form.

[0146] In some embodiments, the pharmaceutical composition comprises a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1), wherein the peptide is cyclized through a disulfide bond formed by the two cysteine residues in the peptide, for example, the two cysteine residues of the peptide are linked via a disulfide bond, or a pharmaceutically acceptable salt or solvate thereof, and optionally a diluent, wherein theAttorney Docket No. T0947.70006WO00 pharmaceutical composition is a lyophilized solid form. In some embodiments, the diluent comprises a zinc salt as described herein.

[0147] In certain embodiments, the pharmaceutical composition comprises a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2, (SEQ ID NO: 1), wherein the peptide is cyclized through a disulfide bond formed by the two cysteine residues in the peptide, for example, the two cysteine residues of the peptide are linked via a disulfide bond or a pharmaceutically acceptable salt or solvate thereof, a bulking agent and a buffering agent, wherein the pharmaceutical composition is a lyophilized solid form.

[0148] In certain embodiments, the pharmaceutical composition comprises a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1), wherein the peptide is cyclized through a disulfide bond formed by the two cysteine residues in the peptide, for example, the two cysteine residues of the peptide are linked via a disulfide bond or a pharmaceutically acceptable salt or solvate thereof, a bulking agent, a buffering agent, wherein the pharmaceutical composition is a lyophilized solid form. In certain embodiments, the lyophilized formulation can be reconstituted with a diluent that includes a zinc salt as described herein.

[0149] In the pharmaceutical composition comprising a lyophilized solid form or the reconstituted lyophilized composition, the peptide of formula (I) can be present in various percentages and concentrations. In some embodiments, the peptide of formula (I) can be present in the composition from about 0.1% to about 80% by weight of the total composition (w / w). In certain embodiments, the peptide of formula (I) in the lyophilized composition is about 25% (w / w) to about 75% (w / w). In one embodiment, the peptide of formula (I) in the lyophilized composition is about 27.5% (w / w) to about 73.7% (w / w). In some embodiments, the peptide of formula (I) in the lyophilized composition is about 10% (w / w), about 15% (w / w), about 20% (w / w), about 25% (w / w), about 27.5% (w / w), about 30% (w / w), about 33% (w / w), about 35%, (w / w) about 38% (w / w), about 40% (w / w), about 45% (w / w), about 47% (w / w), about 50% (w / w), about 54% (w / w), about 57% (w / w), about 60% (w / w), about 62% (w / w), about 65% (w / w), about 67% (w / w), about 70%, (w / w) about 72%(w / w), about 73.7% (w / w), about 75% (w / w), about 80% (w / w), about 85% (w / w), about 47% (w / w), about 90% (w / w), or about 95% (w / w). In one embodiment, the peptide of formula (I) in the lyophilized composition is about 55% (w / w) to about 95% (w / w).

[0150] In the pharmaceutical composition comprising a lyophilized solid form or the lyophilized composition can be reconstituted in a suitable diluent, the peptide of formula (I) can be present inAttorney Docket No. T0947.70006WO00 various percentages and concentrations. In some embodiments, the peptide of formula (I) can be present in the reconstituted lyophilized composition from about 0.1% to about 80% by weight of the total volume of the composition (w / v). In certain embodiments, the peptide of formula (I) in the reconstituted lyophilized composition is about 25% (w / v) to about 75% (w / v). In one embodiment, the peptide of formula (I) in the reconstituted lyophilized composition is about 27.5% (w / v) to about 73.7% (w / v). In some embodiments, the peptide of formula (I) in the reconstituted lyophilized composition is about 27.5% (w / v), about 30% (w / v), about 33% (w / v), about 35%, (w / v) about 38% (w / v), about 40% (w / v), about 45% (w / v), about 47% (w / v), about 50% (w / v), about 54% (w / v), about 57% (w / v), about 60% (w / v), about 62% (w / v), about 65% (w / v), about 67% (w / v), about 70%, (w / v) about 72%(w / v), about 73.7% (w / v), or about 75% (w / v). In one embodiment, the peptide of formula (I) in the reconstituted lyophilized composition is about 55% (w / v) to about 95% (w / v).

[0151] In some embodiments, the bulking agent in the pharmaceutical composition comprising a lyophilized solid form or the lyophilized pharmaceutical composition is about 4% (w / w) to about 70% (w / w). In certain embodiments, the bulking agent in the lyophilized pharmaceutical composition is about 4% (w / w) to about 20% (w / w), about 5% (w / w) to about 30% (w / w), about 4% (w / w) to about 40% (w / w), about 5% (w / w) to about 25% (w / w), about 25% (w / w) to about 39%, (w / w) or about 25% (w / w) to about 69% (w / w). In one embodiment, the bulking agent in the lyophilized pharmaceutical composition is about 4.8% (w / w) to about 38.9% (w / w). In another embodiment, the bulking agent in the lyophilized pharmaceutical composition is about 24.6% (w / w) to about 68.8% (w / w).

[0152] In some embodiments, the buffering agent in the pharmaceutical composition comprising a lyophilized solid form or the lyophilized pharmaceutical composition is 0% (w / w) to about 4% (w / w). In certain embodiments, the buffering agent in the lyophilized pharmaceutical composition is about 0.01% (w / w) to about 4% (w / w). In some embodiments, the buffering agent in the lyophilized pharmaceutical composition is about 0.1% (w / w) to about 2% (w / w), about 0.1% (w / w) to about 3% (w / w), about 1% (w / w) to about 4% (w / w), about 2% (w / w) to about 4% (w / w), or about 1% (w / w) to about 3% (w / w). In one embodiment, the buffering agent in the lyophilized pharmaceutical composition is about 1.7% (w / w) to about 3.7% (w / w). In another embodiment, the buffering agent is absent from the lyophilized pharmaceutical composition. In yet another embodiment, the buffering agent in the lyophilized pharmaceutical composition is about 0.01% (w / w) to about 2.6% (w / w).Attorney Docket No. T0947.70006WO00

[0153] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has greater than about 90% purity (w / w), about 91% purity (w / w), about 92% purity (w / w), about 93% purity (w / w), about 94% purity (w / w), about 95% purity (w / w), about 96% purity (w / w), about 97% purity (w / w), about 98% purity (w / w), or about 99% purity (w / w) after being stored at 5 ºC for about 9 months. In one embodiment, the purity of the lyophilized pharmaceutical composition is greater than about 99% (w / w) after being stored at 5 ºC for about 9 months.

[0154] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has greater than about 90% purity (w / w), about 91% purity (w / w), about 92% purity (w / w), about 93% purity (w / w), about 94% purity (w / w), about 95% purity (w / w), about 96% purity (w / w), about 97% purity (w / w), about 98% purity (w / w), or about 99% purity (w / w) after being stored at 5 ºC for about 18 months. In one embodiment, the purity of the lyophilized pharmaceutical composition is greater than about 99% (w / w) after being stored at 5 ºC for about 18 months.

[0155] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has greater than about 90% purity (w / w), about 91% purity (w / w), about 92% purity (w / w), about 93% purity (w / w), about 94% purity (w / w), about 95% purity (w / w), about 96% purity (w / w), about 97% purity (w / w), about 98% purity (w / w), or about 99% purity (w / w) after being kept at 5 ºC for about 24 months. In one embodiment, the purity of the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition is greater than about 99% purity (w / w) after being kept at 5 ºC for about 24 months.

[0156] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has greater than about 90% purity (w / w), about 91% purity (w / w), about 92% purity (w / w), about 93% purity (w / w), about 94% purity (w / w), about 95% purity (w / w), about 96% purity (w / w), about 97% purity (w / w), about 98% purity (w / w), or about 99% purity (w / w) after being stored at 25 ºC for about 9 months. In one embodiment, the purity of the lyophilized pharmaceutical composition is greater than about 99% (w / w) after being stored at 25 ºC for about 9 months.

[0157] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has greater than about 90% purityAttorney Docket No. T0947.70006WO00 (w / w), about 91% purity (w / w), about 92% purity (w / w), about 93% purity (w / w), about 94% purity (w / w), about 95% purity (w / w), about 96% purity (w / w), about 97% purity (w / w), about 98% purity (w / w), or about 99% purity (w / w) after being stored at 25 ºC for about 18 months. In one embodiment, the purity of the lyophilized pharmaceutical composition is greater than about 99% (w / w) after being stored at 25 ºC for about 18 months.

[0158] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has greater than about 90% purity (w / w), about 91% purity (w / w), about 92% purity (w / w), about 93% purity (w / w), about 94% purity (w / w), about 95% purity (w / w), about 96% purity (w / w), about 97% purity (w / w), about 98% purity (w / w), or about 99% purity (w / w) after being kept at 25 ºC for about 24 months. In one embodiment, the purity of the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition is greater than about 98% (w / w) after 24-month of storage at 25 ºC.

[0159] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has less than about 5% (w / w), about 4.5% (w / w), about 4% (w / w), about 3.5% (w / w), about 3% (w / w), about 2.5% (w / w), about 2% (w / w), about 1.5% (w / w), about 1.4% (w / w), about 1.3% (w / w), about 1.2% (w / w), about 1.1% (w / w), about 1% (w / w), about 0.9% (w / w), about 0.8% (w / w), about 0.7% (w / w), about 0.6% (w / w), about 0.5% (w / w), about 0.4% (w / w), about 0.3% (w / w), about 0.2% (w / w), or about 0.1% (w / w) of the ASP-related degradant after being kept at 5 ºC for about 18 months. In one embodiment, the ASP-related degradant is less than about 0.8% (w / w) after being stored at 5 ºC for about 18 months.

[0160] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has less than about 0.8% (w / w) or about 0.7% (w / w) of ASP-related degradant after being kept at 5 ºC for about 24 months. In one embodiment, the ASP-related degradant is less than about 0.7% (w / w) after being stored at 5 ºC for about 24 months.

[0161] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has less than about 1.7% (w / w), about 1.6% (w / w), about 1.5% (w / w), about 1.4% (w / w), about 1.3% (w / w), about 1.2% (w / w), about 1.1% (w / w), about 1% (w / w), or about 0.9% (w / w), of ASP-related degradant after being kept at 25 ºC for about 18 months. In one embodiment, the ASP-related degradant is less than about 0.9% (w / w) after being stored at 25 ºC for about 18 months.Attorney Docket No. T0947.70006WO00

[0162] In some embodiments, the pharmaceutical composition comprising a lyophilized solid form, or the lyophilized pharmaceutical composition as described herein has less than about 1.2% (w / w), about 1.1% (w / w), about 1% (w / w), or about 0.9% (w / w), of ASP-related degradant after being kept at 25 ºC for about 24 months. In one embodiment, the ASP-related degradant is less than about 0.9% (w / w) after being stored at 25 ºC for about 24 months.

[0163] In some embodiments, the disclosure provides a lyophilized pharmaceutical composition, wherein the composition has at least or greater than 98.7% (w / w) or 98.9% purity (w / w) after storage at 5 ºC for about 24 months. In certain embodiments, the lyophilized pharmaceutical composition has at least or greater than 98.9% purity (w / w) after storage at 5 ºC for about 18 months. In certain embodiments, the lyophilized pharmaceutical composition has at least or greater than 99% purity (w / w) after storage at 5 ºC for about 12 months.

[0164] In some embodiments, the disclosure provides a lyophilized pharmaceutical composition, wherein the composition has at least or greater than 97.2% (w / w) or 97.5% purity (w / w) after storage at 25 ºC for about 24 months. In certain embodiments, the lyophilized pharmaceutical composition has at least or greater than 97.5% (w / w) or 97.9% purity (w / w) after storage at 25 ºC for about 18 months. In certain embodiments, the lyophilized pharmaceutical composition has at least or greater than 98.2% (w / w) or 98.6% purity (w / w) after storage at 25 ºC for about 12 months.

[0165] In some embodiments, the disclosure provides a lyophilized pharmaceutical composition, wherein the composition has less than 0.7% (w / w), 0.8% (w / w), 0.9% (w / w), 1.1% (w / w) or 1.2% ASP related degradant after storage at 5 ºC for about 24 months. In certain embodiments, the lyophilized pharmaceutical composition has less than 0.8% (w / w), 1.3% (w / w) or 1.5% (w / w) ASP related degradant after storage at 5 ºC for about 18 months. In certain embodiments, the lyophilized pharmaceutical composition has less than 0.7% (w / w) ASP related degradant after storage at 5 ºC for about 12 months.

[0166] In some embodiments, the disclosure provides a lyophilized pharmaceutical composition, wherein the composition has less than 0.9% (w / w), 1.1% (w / w) or 1.2% (w / w) ASP related degradant after storage at 25 ºC for about 24 months. In certain embodiments, the lyophilized pharmaceutical composition has less than 0.9% (w / w), 1.1% (w / w) or 1.2% (w / w) ASP related degradant after storage at 25 ºC for about 18 months. In certain embodiments, the lyophilized pharmaceutical composition has less than 0.9% (w / w) ASP related degradant after storage at 25 ºC for about 12 months.Attorney Docket No. T0947.70006WO00

[0167] In another aspect, the disclosure provides a pharmaceutical composition suitable for subcutaneous administration. The pharmaceutical composition comprises a peptide having the amino acid sequence Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2, (SEQ ID NO: 1) for example a peptide of formula (I), or a pharmaceutically acceptable salt thereof, a bulking agent, a buffering agent and a diluent comprising a zinc salt, wherein the two cysteine residues of the peptide are linked via a disulfide bond. Thus, the peptide is cyclized through a disulfide bond formed by the two cysteine residues in the peptide. In one embodiment, the pharmaceutical composition is a reconstituted composition, which is in a liquid form. In one embodiment, the reconstituted pharmaceutical composition is a homogeneous aqueous solution.

[0168] In some embodiments, the reconstituted pharmaceutical composition is in a liquid form which is formed by mixing a lyophilized solid form as described herein with a diluent comprising a zinc salt to form a solution, wherein the lyophilized solid form comprises a peptide having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent and a buffering agent, wherein the two cysteine residues of the peptide are linked via a disulfide bond. In certain embodiments, the lyophilized solid form as described herein and the diluent including a zinc salt are dissolved in the solvent to form a homogenous solution.

[0169] In some embodiments, the reconstituted pharmaceutical composition provided herein further comprises a pH adjuster and a solvent. As such, the disclosure provides a liquid pharmaceutical composition comprising a peptide having the amino acid sequence of Isovaleric acid- DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent, a buffering agent and a diluent comprising a zinc salt, wherein the two cysteine residues of the peptide are linked via a disulfide bond.

[0170] In some embodiments, the pharmaceutical composition in a liquid form as described herein is suitable for subcutaneous administration to a subject.

[0171] In some embodiments of the pharmaceutical composition described herein, for example, the reconstituted liquid pharmaceutical composition, the dosage of the peptide is from about 1 mg to about 100 mg, about 1 mg to about 75 mg, about 1 mg to about 75 mg, or about 10 mg to about 60 mg in the composition.

[0172] In some embodiments, the peptide of formula (I) in the pharmaceutical composition as described herein is about 1 mg to about 10 mg, about 5 mg to about 20 mg, about 10 mg to about 30Attorney Docket No. T0947.70006WO00 mg, about 10 mg to about 60 mg, about 10 mg to about 50 mg, about 15 mg to about 70 mg, about 10 mg to about 75 mg, about 20 mg to about 60 mg, about 30 mg to about 80 mg, about 45 mg to about 80 mg, about 40 mg to about 75 mg, or about 10 mg to about 80 mg.

[0173] In some embodiments, the peptide of formula (I) in the pharmaceutical composition as described herein is about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 15 mg, about 16 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, or about 100 mg.

[0174] In certain embodiments, the peptide of formula (I) in the composition or the dosage of the peptide is about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 45 mg, about 60 mg, about 75 mg, about 80 mg, about 90 mg, or about 100 mg.

[0175] In one embodiment, the peptide in the composition or the dosage of the peptide is about 10 mg.

[0176] In one embodiment, the peptide in the composition or the dosage of the peptide is about 15 mg.

[0177] In one embodiment, the peptide in the composition or the dosage of the peptide is about 20 mg.

[0178] In one embodiment, the peptide in the composition or the dosage of the peptide is about 30 mg.

[0179] In one embodiment, the peptide in the composition or the dosage of the peptide is about 40 mg.Attorney Docket No. T0947.70006WO00

[0180] In one embodiment, the peptide in the composition or the dosage of the peptide is about 40 mg.

[0181] In one embodiment, the peptide in the composition or the dosage of the peptide is about 60 mg.

[0182] In one embodiment, the peptide in the composition or the dosage of the peptide is about 70 mg.

[0183] In one embodiment, the peptide in the composition or the dosage of the peptide is about 80 mg.

[0184] In one embodiment, the peptide in the composition or the dosage of the peptide is about 90 mg.

[0185] In one embodiment, the peptide in the composition or the dosage of the peptide is about 100 mg.

[0186] In some embodiments, the peptide of formula (I) in the reconstituted pharmaceutical composition as described herein is about 0.1% (w / v) to about 20% (w / v); about 0.2% (w / v) to about 15% (w / v); about 0.3% (w / v) to about 15% (w / v); about 0.4% (w / v) to about 15% (w / v); about 0.5% (w / v) to about 15% (w / v); about 0.6% (w / v) to about 15% (w / v); about 0.7% (w / v) to about 15% (w / v); about 0.8% (w / v) to about 15% (w / v); about 0.9% (w / v) to about 15% (w / v); about 1% (w / v) to about 15% (w / v); about 1% (w / v) to about 12% (w / v); about 2% (w / v) to about 12% (w / v); about 3% (w / v) to about 15% (w / v); about 4% (w / v) to about 12% (w / v); about 1% (w / v) to about 6% (w / v), or about 6% (w / v) to about 12% (w / v). The unit “w / v” means by weight per volume. In one embodiment, the peptide in the composition is about 2% (w / v) to about 12% (w / v). In certain instances, w / v means mg / mL.

[0187] In some embodiments, the peptide in the reconstituted pharmaceutical composition is about 1% (w / v), about 2% (w / v), about 3% (w / v), about 4% (w / v), about 5% (w / v), about 6% (w / v), about 7% (w / v), about 8% (w / v), about 9% (w / v), about 10% (w / v), about 11% (w / v), about 12% (w / v), about 13% (w / v), about 14% (w / v), about 15% w / v, about 16% (w / v), about 17% (w / v), about 18% (w / v), about 19% (w / v), about 20% (w / v), about 21% (w / v), about 22% (w / v), about 23% (w / v), about 24% (w / v), about 25% (w / v), about 26% (w / v), about 27% (w / v), about 28% (w / v), about 29% (w / v), about 30% (w / v), about 31% (w / v), about 32% (w / v), about 33% (w / v), about 34% (w / v), about 35% w / v, about 36% (w / v), about 37% (w / v), about 38% (w / v), about 39% (w / v), about 40% (w / v), about 41% (w / v), about 42% (w / v), about 43% (w / v), about 44% (w / v), about 45% (w / v),Attorney Docket No. T0947.70006WO00 about 46% (w / v), about 47% (w / v), about 48% (w / v), about 49% (w / v), about 50% (w / v), about 51% (w / v), about 52% (w / v), about 53% (w / v), about 54% (w / v), about 55% w / v (w / v), about 56% (w / v), about 57% (w / v), about 58% (w / v), about 59% (w / v), about 60% (w / v), about 61% (w / v), about 62% (w / v), about 63% (w / v), about 64% (w / v), about 65% (w / v), about 66% (w / v), about 67% (w / v), about 68% (w / v), about 69% (w / v), about 70% (w / v), about 71% (w / v), about 72% (w / v), about 73% (w / v), about 74% (w / v), or about 75% w / v.

[0188] In some embodiments, the peptide in the composition is about 1% (w / v) to about 6% (w / v). In certain instances, w / v is mg / mL.

[0189] In some embodiments, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is from about 1 mM to about 30 mM. In one embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 4 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 6 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 8 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 12 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 8 mM to about 18 mM.

[0190] In some embodiments, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15m, MM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM. In other embodiments, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 4 mM, about 6 mM, about 8 mM, about 12 mM, about 18 mM, about 24 mM, or about 25 mM. In one embodiment, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 4.1 mM, about 6.15 mM, about 8.2 mM, about 12.29 mM, about 18.44 mM, or about 24.59 mM. In one embodiment, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 4.1 mM – about 24.59 mM.Attorney Docket No. T0947.70006WO00

[0191] In the liquid pharmaceutical compositions as described herein (SLF or reconstituted Lyo), the peptide of formula (I) (Compound 1) can be present in various percentages and concentrations. In some embodiments, the peptide of formula (I) can be present in the liquid composition from about 0.01% to about 30% by weight of the total volume (w / v). In certain embodiments, the peptide of formula (I) in the liquid pharmaceutical composition is about 0.01% (w / v) to about 1% (w / v). In certain embodiments, the peptide of formula (I) in the liquid pharmaceutical composition is about 0.1% (w / v) to about 30% (w / v). In certain embodiments, the peptide of formula (I) in the liquid pharmaceutical composition is about 0.1% (w / v) to about 12% (w / v). In one embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 0.2% (w / v) to about 4.0% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 1% (w / v) to about 6% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 2% (w / v) to about 4.5% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 1% (w / v) to about 3% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 0.1% (w / v) to about 6% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 1% (w / v) to about 7.5% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 1% (w / v) to about 10% (w / v).

[0192] In some embodiments, the peptide of formula (I) in the liquid pharmaceutical composition as described herein is about 0.1% (w / v) to about 20% (w / v); about 0.2% (w / v) to about 15% (w / v); about 0.3% (w / v) to about 15% (w / v); about 0.4% (w / v) to about 15% (w / v); about 0.5% (w / v) to about 15% (w / v); about 0.6% (w / v) to about 15% (w / v); about 0.7% (w / v) to about 15% (w / v); about 0.8% (w / v) to about 15% (w / v); about 0.9% (w / v) to about 15% (w / v); about 1% (w / v) to about 15% (w / v); about 1% (w / v) to about 12% (w / v); about 2% (w / v) to about 12% (w / v); about 3% (w / v) to about 15% (w / v); about 4% (w / v) to about 12% (w / v); about 1% (w / v) to about 6% (w / v), or about 6% (w / v) to about 12% (w / v). The unit “w / v” means by weight per volume. In one embodiment, the peptide in the composition is about 0.1% (w / v) to about 12% (w / v). In certain instances, w / v means mg / mL.

[0193] In some embodiments, the peptide of formula (I) in the liquid pharmaceutical composition is about 0.1% (w / v), about 0.2% (w / v), about 0.3% (w / v), about 0.4%, (w / v) about 0.5% (w / v), about 0.6% (w / v), about 0.7% (w / v), about 0.8% (w / v), about 0.9% (w / v), about 1% (w / v), about 1.2%Attorney Docket No. T0947.70006WO00 (w / v), about 1.4% (w / v), about 1.5% (w / v), about 1.8% (w / v), about 2% (w / v), about 2.2%, (w / v) about 2.5%(w / v), about 2.7% (w / v), about 3% (w / v), about 3.2% (w / v), about 3.5% (w / v), about 3.7% (w / v), about 3.8% (w / v), about 4% (w / v), about 4.2% (w / v), about 4.5% (w / v), about 4.7% (w / v), about 4.9% (w / v), about 5% (w / v), about 5.2% (w / v), about 5.4% (w / v), about 5.5% (w / v), about 5.7% (w / v), about 5.9% (w / v), about 6% (w / v), about 6.3% (w / v), about 6.5% (w / v), about 6.8% (w / v), about 7% (w / v), about 7.3% (w / v), about 7.5% (w / v), about 7.8% (w / v), about 8% (w / v), about 8.3% (w / v), about 8.5% (w / v), about 8.8% (w / v), about 9% (w / v), about 9.3% (w / v), about 9.5% (w / v), about 9.8% (w / v), about 10% (w / v), about 10.3 % (w / v), about 10.5% (w / v), about 10.8% (w / v), about 11% (w / v), about 11.3% (w / v), about 11.5% (w / v), about 11.8% (w / v), about 12% (w / v), about 13% (w / v), about 14% (w / v), about 15% (w / v), about 16% (w / v), about 17% (w / v), about 18% (w / v), about 19% (w / v), about 20% (w / v), about 21% (w / v), about 22% (w / v), about 23% (w / v), about 24% (w / v), about 25% (w / v), about 26% (w / v), about 27% (w / v), about 28% (w / v), about 29% (w / v), or about 30% (w / v). In one embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 2% (w / v). In another embodiment, the peptide of formula (I) in the liquid pharmaceutical composition is about 4.5% (w / v).

[0194] In some embodiments of the pharmaceutical composition described herein, the dosage of peptide of formula (I) (Compound 1) is from about 1 mg to about 80 mg, about 1 mg to about 75 mg, about 1 mg to about 60 mg, about 10 mg to about 75 mg, or about 10 mg to about 60 mg in the liquid formulation.

[0195] In some embodiments, the peptide of formula (I) in the liquid pharmaceutical composition as described herein is about 1 mg to about 10 mg, about 5 mg to about 20 mg, about 10 mg to about 30 mg, about 10 mg to about 60 mg, about 10 mg to about 50 mg, about 15 mg to about 70 mg, about 10 mg to about 75 mg, about 20 mg to about 60 mg, about 30 mg to about 80 mg, about 45 mg to about 80 mg, about 40 mg to about 75 mg, or about 10 mg to about 80 mg.

[0196] In some embodiments, the peptide of formula (I) in the liquid pharmaceutical composition as described herein is about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 15 mg, about 16 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, 45 mg, about 46 mg, about 47 mg, about 48Attorney Docket No. T0947.70006WO00 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, or about 80 mg.

[0197] In certain embodiments, the peptide of formula (I) in the liquid pharmaceutical composition or the dosage of the peptide is about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 45 mg or about 60 mg.

[0198] In one embodiment, the peptide in the liquid pharmaceutical composition or the dosage of the peptide is about 10 mg.

[0199] In one embodiment, the peptide in the liquid pharmaceutical composition or the dosage of the peptide is about 15 mg.

[0200] In one embodiment, the peptide in the liquid pharmaceutical composition or the dosage of the peptide is about 20 mg.

[0201] In one embodiment, the peptide in the liquid pharmaceutical composition or the dosage of the peptide is about 30 mg.

[0202] In one embodiment, the peptide in the liquid pharmaceutical composition or the dosage of the peptide is about 45 mg.

[0203] In one embodiment, the peptide in the liquid pharmaceutical composition or the dosage of the peptide is about 60 mg.

[0204] In some embodiments, the concentration of the peptide of formula (I) (Compound 1) in the liquid pharmaceutical composition is from about 1 mM to about 30 mM. In one embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 4 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 6 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 8 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 12 mM to about 25 mM. In another embodiment, the concentration of the peptide of formula (I) in the pharmaceutical composition is from about 8 mM to about 18 mM.Attorney Docket No. T0947.70006WO00

[0205] In some embodiments, the concentration of the peptide of formula (I) in the liquid pharmaceutical composition is about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15m, MM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, or about 30 mM.

[0206] In other embodiments, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 4 mM, about 6 mM, about 8 mM, about 12 mM, about 18 mM, about 24 mM, or about 25 mM. In one embodiment, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 4.1 mM, about 6.15 mM, about 8.2 mM, about 12.29 mM, about 18.44 mM, or about 24.59 mM. In one embodiment, the concentration of the peptide of formula (I) in the reconstituted pharmaceutical composition is about 4.1 mM – about 24.59 mM.

[0207] In some embodiments of the liquid pharmaceutical composition as described herein, the zinc salt can be present in the composition from about 0.0001% w / v to about 5% w / v.

[0208] In some embodiments, the zinc salt selected from zinc chloride, zinc acetate, zinc carbonate and zinc oxide. In one embodiment, the zinc salt is zinc acetate. In another embodiment, zinc salt used herein is zinc chloride.

[0209] In other embodiments, the formulations herein further comprise a magnesium salt, such as MgCl2and / or a calcium salt, such as CaCl2.

[0210] In certain embodiments, the zinc salts used in the liquid pharmaceutical composition described herein can be zinc salts of an amino acid. Suitable amino acids that can be used to form a zinc salt include all naturally occurring amino acids and non-naturally occurring amino acids.

[0211] In some embodiments, the amino acids that can form a zinc salt include alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine, each of which can be either an L-amino acid or D-amino acid.

[0212] The zinc salts used in the liquid pharmaceutical composition as described herein can be one or more zinc salts. In some embodiments, the zinc salts used in the compositions can be a single zinc salt or a combination of two or more zinc salts.Attorney Docket No. T0947.70006WO00

[0213] In certain embodiments, zinc salt used in the liquid pharmaceutical composition as described herein is a zinc salt of aspartic acid, glutamic acid, glutamine, or asparagine, or a combination thereof.

[0214] In some embodiments, the zinc salt in the liquid pharmaceutical composition as described herein can be from about 0.0001% (w / v) to about 5% (w / v). In one embodiment, the zinc salt present in the liquid pharmaceutical composition as described herein is from about 0.01% w / v to about 3.3% w / v or about 0.01% w / v to about 5% w / v. In another embodiment, the zinc salt present in the liquid pharmaceutical composition as described herein is from about 0.1% w / v to about 2% w / v or about 0.09% w / v to about 1% w / v. In another embodiment, the zinc salt present in the liquid pharmaceutical composition as described herein is from about 0.2% w / v to about 4% w / v. In another embodiment, the zinc salt present in the liquid pharmaceutical composition as described herein is about 0.1% w / v to about 0.7% w / v.

[0215] In some embodiments, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.01% w / v, about 0.02% w / v, about 0.03% w / v, about 0.04% w / v, about 0.05% w / v, about 0.06% w / v, about 0.07% w / v, about 0.08% w / v, about 0.09% w / v, about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1% w / v, about 1.2% w / v, about 1.5% w / v, about 1.6% w / v, about 1.8% w / v, about 1.9% w / v, about 2% w / v, about 2.2% w / v, about 2.5% w / v, about 2.6% w / v, about 2.8% w / v, about 2.9% w / v, about 3% w / v, about 3.1% w / v, about 3.2% w / v, about 3.3% w / v, about 3.4% w / v, about 3.5% w / v, about 3.8% w / v, about 4% w / v, about 4.2% w / v, about 4.5% w / v, or about 5% w / v. In certain instances, w / v as described herein means mg / mL.

[0216] In one embodiment, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.22% w / v.

[0217] In one embodiment, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.11% w / v.

[0218] In one embodiment, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.084% w / v.

[0219] In one embodiment, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.5% w / v.

[0220] In one embodiment, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.25% w / v.Attorney Docket No. T0947.70006WO00

[0221] In one embodiment, the zinc salt in the liquid pharmaceutical composition as described herein is about 0.19% w / v.

[0222] In some embodiments, the concentration of a zinc salt present in the liquid pharmaceutical composition as described herein is about 0.1 mM to about 10 mM, about 0.1 mM to about 5 mM, about 0.2 mM to about 0.6 mM, about 0.5 mM to about 4 mM, about 1 mM to about 5 mM, about 2 mM to about 6 mM, about 3 mM to about 5 mM, about 3 mM to about 5 mM or about 0.8 mM to about 5 mM.

[0223] In one embodiment, the concentration of a zinc salt present in the liquid pharmaceutical composition as described herein is about 4.9 mM.

[0224] In some embodiments, the concentration of a zinc salt present in the liquid pharmaceutical composition as described herein is about 1 mg / mL to about 5 mg / mL, about 2.5 mg / mL to about 3.5 mg / mL, or about 2.8 mg / mL to about 3.2 mg / mL.

[0225] In other embodiments, the concentration of a zinc salt in the liquid pharmaceutical composition as described herein is about 2.8 mg / mL to about 3.0 mg / mL.

[0226] In other embodiments, the concentration of a zinc salt in the liquid pharmaceutical composition is about 2.85 mg / mL to about 2.95 mg / mL.

[0227] In one embodiment, the concentration of a zinc salt in the liquid pharmaceutical composition as described herein is about 2.9 mg / mL.

[0228] In some embodiments of the liquid pharmaceutical compositions as described herein, the zinc salt and the peptide (Compound 1) have a molar ratio from about 0.5:1 to about 3:1; about 0.75:1 to about 3:1; about 0.75:1 to about 2:1; about 0.05:1 to about 2:1; about 0.05 to about 1.6:1; about 0.1:1 to about 2:1; about 0.1:1 to about 1.5:1; about 0.1:1 to about 1.2:1; about 0.18 to about 0.36; about 0.2:1 to about 1.2:1; about 0.2:1 to about 0.6:1; about 0.3:1 to about 1.2:1; about 0.4:1 to about 1.2:1; or about 1:1 to about 2:1.

[0229] In one embodiment of the pharmaceutical compositions as described herein, the zinc salt and the peptide have a molar ratio from about 1:1 to about 2:1.

[0230] In one embodiment of the pharmaceutical compositions as described herein, the zinc salt and the peptide have a molar ratio from about 0.75:1 to about 3:1.

[0231] In one embodiment of the pharmaceutical compositions as described herein, the zinc salt and the peptide have a molar ratio from about 0.2:1 to about 1.2:1.Attorney Docket No. T0947.70006WO00

[0232] In one embodiment, the zinc salt and peptide have a molar ratio of about 2:1 in the pharmaceutical compositions as described herein.

[0233] In one embodiment, the zinc salt and peptide have a molar ratio of about 1:1 in the pharmaceutical compositions as described herein.

[0234] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.75:1 in the pharmaceutical compositions as described herein.

[0235] In one embodiment, the zinc salt and peptide have a molar ratio of about 1.2:1 in the pharmaceutical compositions as described herein.

[0236] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.4:1 in the pharmaceutical compositions as described herein.

[0237] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.3:1 in the pharmaceutical compositions as described herein.

[0238] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.2:1 in the pharmaceutical compositions as described herein.

[0239] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.5:1 in the pharmaceutical compositions as described herein.

[0240] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.1:1 in the pharmaceutical compositions as described herein.

[0241] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.6:1 in the pharmaceutical compositions as described herein.

[0242] Various buffering agents can be used in liquid pharmaceutical compositions as described herein. In some embodiments, the buffering agent is selected from sodium acetate, sodium acetate trihydrate, phosphates, ammonium sulfate, ammonium hydroxide, arginine, aspartic acid, benzene sulfonic acid, sodium benzoate / benzoic acid, boric acid / sodium boric acid salt, sodium carbonate, carbon dioxide, citrate, citric acid, diethanolamine, glucono delta lactone, glycine / glycine HCl, histidine / histidine HCl, hydrochloric acid, hydrobromic acid, lysine (L), maleic acid, meglumine, methanesulfonic acid, monoethanolamine, sodium hydroxide, succinate sodium / disodium, sulfuric acid, tromethamine (Tris), EDTA, tartaric acid, Bis-Tris, MOPS, MES, aspartate, ascorbate, 2,2′-[(2- amino-2-oxoethyl)azanediyl]diacetic acid (ADA), borate, PBS, sodium citrate, TRIS, piperazine- N,N'-bis(ethanesulfonic acid) (PIPES), HEPES, HEPES-Na, HEPBS, MOPS, MOBS, tartrate,Attorney Docket No. T0947.70006WO00 succinate, malonate, gluconate, aspartate, histidine, TABS, TES, BSA, TES, TAPS, EPPS, MES, CHES, CAPS or CABS.

[0243] In some embodiments, the buffering agent is sodium acetate, sodium phosphate, sodium citrate, or TRIS.

[0244] In one embodiment, the buffering agent is sodium acetate. In another embodiment, the buffering agent is sodium acetate trihydrate.

[0245] In some embodiments, the buffering agent in the liquid pharmaceutical compositions as described herein is from about 0.01% (w / v) to about 1% (w / v), 0.01% (w / v) to about 0.068% (w / v), about 0.05 (w / v) to about 0.1% (w / v), about 0.01% (w / v) to about 1% (w / v), or from 0 to about 0.68% w / v.

[0246] In some embodiments, the buffering agent in the pharmaceutical compositions as described herein is about 0.0001% w / v to about 0.68% w / v or about 0.01% w / v to about 0.68% w / v.

[0247] In some embodiments, the buffering agent in the liquid pharmaceutical compositions as described herein is about 0.01% w / v, about 0.02% w / v, about 0.03% w / v, about 0.05% w / v, about 0.06% w / v, about 0.068% w / v, about 0.07% w / v, about 0.08% w / v, about 0.09% w / v, about 0.1% w / v, about 0.164% w / v, about 0.2% w / v, about 0.272% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.68% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1% w / v, about 1.2% w / v, about 1.3% w / v, about 1.4% w / v, about 1.5% w / v, about 1.6% w / v, about 1.7% w / v, about 1.8% w / v, about 1.9% w / v or about 2% w / v.

[0248] In one embodiment, the buffering agent in the liquid pharmaceutical compositions as described herein is about 0.164% w / v. In another embodiment, the buffering agent in the liquid pharmaceutical compositions as described herein is about 0.272% w / v.

[0249] In some embodiments, the buffering agent in the liquid pharmaceutical compositions as described herein, the buffering agent is about 1 mg / mL to about 5 mg / mL or about 2 mg / mL to about 4 mg / mL.

[0250] In one embodiment, the buffering agent in the liquid pharmaceutical compositions as described herein is about 2.72 mg / mL.

[0251] In one embodiment, the buffering agent in the liquid pharmaceutical compositions as described herein is about 4.08 mg / mL.

[0252] The liquid pharmaceutical compositions as described herein can further include a stabilizing agent or a stabilizer. The stabilizing agents or stabilizers used in the liquid composition as describedAttorney Docket No. T0947.70006WO00 herein include, but are not limited to, amino acids, disaccharides, polysaccharides, polyols, surfactants, buffers, antioxidants, polymers, salts, chelating agents, inclusion complexes, metal salts, and combinations thereof.

[0253] Exemplary amino acids used as stabilizing agents or stabilizers include L-glycine, L- arginine, L-alanine, L-proline, L-aspartic acid, L-glutamic acid, L-lysine, L-methionine, L-serine, L- threonine, L-asparagine, L-histidine, L-cystine and combinations thereof.

[0254] Exemplary disaccharides or polysaccharides used as stabilizing agents include sucrose, trehalose, lactose, maltose, lactulose, cellobiose, chitobiose, and dextran. In some embodiments, the stabilizing agent used in the liquid composition as described herein is sucrose, trehalose, lactose or a combination thereof.

[0255] Exemplary polyols used as stabilizing agents or stabilizers include sorbitol, mannitol, glycerol, erythritol, isomalt, lactitol, maltitol, and xylitol. In some embodiments, the stabilizing agent is sorbitol, mannitol, or glycerol.

[0256] Exemplary surfactants used as stabilizing agents or stabilizers include poloxamer 407, poloxamer 188*, polysorbate 80, sodium lauryl sulfate, and Miglyol 810.

[0257] Exemplary buffering agents used as stabilizing agents or stabilizers include sodium acetate, sodium phosphate, PBS, sodium citrate, TRIS, piperazine-N,N'-bis(ethanesulfonic acid) (PIPES), HEPES, HEPES-Na, HEPBS, or MOPS, MOBS, tartrate, succinate, malonate, gluconate, aspartate, histidine, TABS, TES, BSA, TES, TAPS, EPPS, MES, CHES, CAPS, and CABS. In some embodiments, the stabilizing agent is a buffering agent selected from sodium acetate, sodium phosphate, sodium citrate, or TRIS.

[0258] Exemplary antioxidants used as stabilizing agents or stabilizers include L-ascorbic acid, and glutathione.

[0259] Exemplary polymers used as stabilizing agents or stabilizers include polyethylene glycol having molecular weight between 100 to 10000, PEG 3350, PEG 400, and Carboxyl MC. In one embodiment, the stabilizer is polyethylene glycol having a molecular weight between 1000 to 10000. In one embodiment, the stabilizer is PEG3350.

[0260] Exemplary salt as a stabilizer includes NaCl.

[0261] Exemplary chelating agent as a stabilizer includes EDTA.

[0262] Exemplary inclusion complexes used as stabilizing agents or stabilizers include Captisol, Kleptose HPB, Kleptose HP.Attorney Docket No. T0947.70006WO00

[0263] Exemplary metal salts used as stabilizing agents or stabilizers include Zinc chloride, zinc acetate, magnesium chloride, and calcium chloride.

[0264] In one embodiment, the stabilizing agent or stabilizer in the liquid composition as described herein is PEG3350.

[0265] In some embodiments, the stabilizing agent or stabilizer in the liquid composition as described herein is from 0 to about 5% w / v or from about 0.01% w / v to about 3.6% w / v. in one embodiment, the stabilizing agent or stabilizer in the liquid composition as described herein is about 1.8% w / v.

[0266] The liquid pharmaceutical compositions as described herein can further include a tonicity agent or tonicity adjuster. Examples of tonicity agents include, but are not limited to, salts and polyols. Exemplary salts as a tonicity agent include sodium acetate, sodium phosphate, sodium citrate, TRIS, and sodium chloride. In one embodiment the salt is sodium chloride.

[0267] Exemplary polyols as a tonicity agent include sorbitol, mannitol, glycerol, erythritol, isomalt, lactitol, maltitol and xylitol. In one embodiment, the tonicity agent is sorbitol.

[0268] In some embodiments, the tonicity agent in the liquid composition is 0 to about 6% w / v or 0 to about 5.5% w / v. In certain embodiments, the tonicity agent in the liquid composition is from about 1% w / v to about 5% w / v, about 0.9% w / v to about 4.9% w / v, about 0.5% w / v to about 5% w / v, about 3% w / v to about 5% w / v, or about 3.1% w / v to about 5.5% w / v.

[0269] In certain embodiments, the tonicity agent in the liquid composition is from about 0.9% w / v to about 4.9% w / v. In other embodiments, the tonicity agent in the liquid composition is from about 3.1% w / v to about 5.5% w / v.

[0270] In some embodiments, the tonicity agent in the liquid composition is about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1% w / v, about 1.2% w / v, about 1.3% w / v, about 1.5% w / v, about 1.6% w / v, about 1.8% w / v, about 1.9% w / v, about 2.0% w / v, about 2.1% w / v, about 2.2% w / v, about 2.3% w / v, about 2.4% w / v, about 2.5% w / v, about 2.6% w / v, about 2.7% w / v, about 2.8% w / v, about 2.9% w / v, about 3.0% w / v, about 3.1% w / v, about 3.2% w / v, about 3.3% w / v, about 3.4% w / v, about 3.5% w / v, about 3.6% w / v, about 3.7% w / v, about 3.8% w / v, about 3.9% w / v, about 4% w / v, about 4.1% w / v, about 4.2% w / v, about 4.3% w / v, about 4.4% w / v, about 4.5% w / v, about 4.6% w / v, about 4.7% w / v, about 4.8% w / v, about 4.9% w / v, about 5% w / v, about 5.1%Attorney Docket No. T0947.70006WO00 w / v, about 5.2% w / v, about 5.3% w / v, about 5.4% w / v, about 5.5% w / v, about 5.6% w / v, about 5.7% w / v, about 5.8% w / v, about 5.9% w / v, or about 6% w / v.

[0271] In some embodiments, the tonicity agent, such as sorbitol in the liquid composition is from about 3.7% w / v to about 5.8% w / v.

[0272] In some embodiments, the tonicity agent, such as sorbitol in the liquid composition is about 3.7% w / v, 3.8% w / v, 3.9% w / v, 4.0% w / v, 4.1% w / v, 4.2% w / v, 4.3% w / v, 4.4% w / v, 4.5% w / v, 4.6% w / v, 4.7% w / v, 4.8% w / v, 4.9% w / v, 5% w / v, 5.1% w / v, 5.2% w / v, 5.3% w / v, 5.4% w / v, 5.5% w / v, 5.6% w / v, 5.7% w / v, 5.8% w / v, or 5.9% w / v.

[0273] The liquid pharmaceutical compositions as described herein can further include a solubilizing agent or a solubilizer. Without being bound by any theory, in some embodiments, at least one solubilizing agent may be added to increase the solubility of the peptide (Compound 1).

[0274] Exemplary solubilizing agents include solvents, inclusion complexes and surfactants.

[0275] In some embodiments, the solubilizing agent is a solvent selected from NMP, DMSO, benzyl benzoate, propylene glycol, polyethylene glycol (300, 400, 600, 3350, 4000), glycerin, benzyl benzoate, oleic acid, castor oil, cottonseed oil, N,N dimethylacetamide, ethanol / ethanol dehydrated, glycerin (glycerol), peanut oil, poppy seed oil, safflower seed oil, sesame oil, soybean oil, and vegetable oil.

[0276] In certain embodiments, the solubilizing agent is an inclusion complex selected from Captisol, Kleptose HPB and Kleptose HP.

[0277] In certain embodiments, the solubilizing agent is a surfactant selected from Miglyol 810 Polyoxyl 35 castor oil (Cremophor EL), Polyoxyl 40 hydrogenated castor oil (Cremophor RH 40), Polyoxyl 60 hydrogenated castor oil (Cremophor RH 60), Polysorbate 20 (Tween 20), Polysorbate 80 (Tween 80), D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), Solutol HS-15 Sorbitan monooleate (Span 20), PEG 300 caprylic / capric glycerides (Softigen 767), PEG 400 caprylic / capric glycerides (Labrasol), PEG 300 oleic glycerides (Labrafil M-1944CS), PEG 300 linoleic glycerides (Labrafil M-2125CS), Polyoxyl 8 stearate (PEG 400 monosterate), Polyoxyl 40 stearate (PEG 1750 monosterate), poloxamer 188, poloxamer 407, and phospholipids.

[0278] The liquid pharmaceutical compositions as described herein can further comprise a pH adjuster, such as acetic acid or glacial acetic acid to allow the pH of the liquid pharmaceutical composition to be adjusted to a value suitable for injection to the patient. A base such as NaOH can also be used in combination with an acid to adjust the pH to a desirable value. In one embodiment,Attorney Docket No. T0947.70006WO00 the pH of the pharmaceutical composition is about 5.3 to about 5.6. In another embodiment, the pH of the reconstituted pharmaceutical composition is about 5.3, about 5.4, about 5.5 or about 5.6.

[0279] In one embodiment, the pH of the liquid pharmaceutical composition is about 5.4.

[0280] The final volume of the liquid pharmaceutical composition suitable for injection to the subject can be from about 0.5 mL or about 1.0 mL, which can be achieved by, for example, adding a solvent such as water to the composition. In some embodiments, the volume of the liquid pharmaceutical compositions can be about 0.5 mL, about 0.6 mL, about 0.7 mL, about 0.8 mL, about 0.9 mL or about 1.0 mL. In one embodiment, the volume of the liquid pharmaceutical compositions as described herein is 1.0 mL.

[0281] In the pharmaceutical composition as described herein, the diluent can comprise various zinc salts. In some embodiments, the zinc salt is zinc maleate, zinc tartrate, zinc carbonate, zinc oxide, zinc hydroxide, zinc mesylate, zinc besylate, zinc chloride, or zinc acetate. In one embodiment, the zinc salt is zinc acetate. In another embodiment, zinc salt used herein is zinc chloride.

[0282] In certain embodiments, the zinc salts used in the pharmaceutical composition described herein can be zinc salts of an amino acid. Suitable amino acids that can be used to form a zinc salt include all naturally occurring amino acids and non-naturally occurring amino acids.

[0283] In some embodiments, the amino acids that can form a zinc salt include alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine, each of which can be either an L-amino acid or D-amino acid.

[0284] The zinc salts used in the pharmaceutical composition as described herein can be one or more zinc salts. In some embodiments, the zinc salts used in the composition can be a single zinc salt or a combination of two or more zinc salts.

[0285] In certain embodiments, the zinc salt used in the pharmaceutical composition as described herein is a zinc salt of aspartic acid, glutamic acid, glutamine, or asparagine, or a combination thereof.

[0286] In some embodiments, the zinc salt present in the pharmaceutical composition as described herein can be from about 0.01% (w / v) to about 5% (w / v). In one embodiment, the zinc salt present in the pharmaceutical composition as described herein is from about 0.01% w / v to about 3.3% w / v. In another embodiment, the zinc salt in the pharmaceutical composition as described herein is from about 0.1% w / v to about 2% w / v or about 0.09% w / v to about 1% w / v.Attorney Docket No. T0947.70006WO00

[0287] In some embodiments, the zinc salt in the pharmaceutical composition as described herein is about 0.01% w / v, about 0.02% w / v, about 0.03% w / v, about 0.04% w / v, about 0.05% w / v, about 0.06% w / v, about 0.07% w / v, about 0.08% w / v, about 0.09% w / v, about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1% w / v, about 1.2% w / v, about 1.5% w / v, about 1.6% w / v, about 1.8% w / v, about 1.9% w / v, about 2% w / v, about 2.2% w / v, about 2.5% w / v, about 2.6% w / v, about 2.8% w / v, about 2.9% w / v, about 3% w / v, about 3.1% w / v, about 3.2% w / v, about 3.3% w / v, about 3.4% w / v, about 3.5% w / v, about 3.8% w / v, about 4% w / v, about 4.2% w / v, about 4.5% w / v, or about 5% w / v. In certain instances, w / v as described herein means mg / mL.

[0288] In one embodiment, the zinc salt present in the pharmaceutical composition as described herein is about 0.2% w / v.

[0289] In one embodiment, the zinc salt present in the pharmaceutical composition as described herein is about 0.09% w / v.

[0290] In some embodiments, the zinc salt present in the pharmaceutical composition as described herein is about 1 mg / mL to about 5 mg / mL.

[0291] In some embodiments, the concentration of a zinc salt present in the reconstituted pharmaceutical composition as described herein is about 0.1 mM to about 10 mM, about 0.1 mM to about 5 mM, about 0.2 mM to about 0.6 mM, about 0.5 mM to about 4 mM, about 1 mM to about 5 mM, about 2 mM to about 6 mM, about 3 mM to about 5 mM, about 3 mM to about 5 mM or about 0.8 mM to about 5 mM.

[0292] In some embodiments, the concentration of a zinc salt present in the reconstituted pharmaceutical composition as described herein is about 0.1 mM, about 0.2 mM, about 0.3 mM, about 0.4 mM, about 0.5 mM, about 0.6 mM, about 0.7 mM, about 0.8 mM, about 0.9 mM, about 1 mM, about 1.1 mM, about 1.2 mM, about 1.3 mM, about 1.4 mM, about 1.5 mM, about 1.6 mM, about 1.7 mM, about 1.8 mM, about 1.9 mM, about 2 mM, about 2.1 mM, about 2.2 mM, about 2.3 mM, about 2.4 mM, about 2.5 mM, about 2.6 mM, about 2.7 mM, about 2.8 mM, about 2.9 mM, about 3.0 mM, about 3.1 mM, about 3.2 mM, about 3.3 mM, about 3.4 mM, about 3.5 mM, about 3.6 mM, about 3.7 mM, about 3.8 mM, about 3.9 mM, about 4.0 mM, about 4.1 mM, about 4.2 mM, about 4.3 mM, about 4.4 mM, about 4.5 mM, about 4.6 mM, about 4.7 mM, about 4.8 mM, about 4.9 mM or about 5.0 mM.Attorney Docket No. T0947.70006WO00

[0293] In one embodiment, the concentration of a zinc salt present in the reconstituted pharmaceutical composition as described herein is about 4.9 mM.

[0294] In some embodiments, the concentration of a zinc salt present in the reconstituted pharmaceutical composition as described herein is about 1 mg / mL to about 5 mg / mL, about 2.5 mg / mL to about 3.5 mg / mL, or about 2.8 mg / mL to about 3.2 mg / mL.

[0295] In other embodiments, the concentration of a zinc salt in the reconstituted pharmaceutical composition as described herein is about 2.8 mg / mL to about 3.0 mg / mL.

[0296] In other embodiments, the concentration of a zinc salt in the reconstituted pharmaceutical composition is about 2.85 mg / mL to about 2.95 mg / mL.

[0297] In one embodiment, the concentration of a zinc salt in the reconstituted pharmaceutical composition as described herein is about 2.9 mg / mL.

[0298] In some embodiments of the pharmaceutical compositions as described herein, the zinc salt and the peptide have a molar ratio from about 0.05:1 to about 2:1; about 0.05 to about 1.6:1; about 0.1:1 to about 2:1; about 0.1:1 to about 1.5: 1; about 0.1:1 to about 1.2:1; about 0.18 to about 0.36; about 0.2:1 to about 1.2:1; about 0.2:1 to about 0.6:1; about 0.3:1 to about 1.2:1; or about 0.4:1 to about 1.2:1.

[0299] In one embodiment of the pharmaceutical compositions as described herein, the zinc salt and the peptide have a molar ratio from about 0.2:1 to about 1.2:1.

[0300] In one embodiment, the zinc salt and peptide have a molar ratio of about 1.2:1 in the pharmaceutical compositions as described herein.

[0301] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.6:1 in the pharmaceutical compositions as described herein.

[0302] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.4:1 in the pharmaceutical compositions as described herein.

[0303] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.3:1 in the pharmaceutical compositions as described herein.

[0304] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.2:1 in the pharmaceutical compositions as described herein.

[0305] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.5:1 in the pharmaceutical compositions as described herein.Attorney Docket No. T0947.70006WO00

[0306] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.1:1 in the pharmaceutical compositions as described herein.

[0307] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.6:1 in the pharmaceutical compositions as described herein.

[0308] In one embodiment, the zinc salt and peptide have a molar ratio of about 0.05:1 in the pharmaceutical compositions as described herein. In another embodiment, the zinc salt and peptide have a molar ratio of about 0.06:1 in the pharmaceutical compositions as described herein. In another embodiment, the zinc salt and peptide have a molar ratio of about 0.07:1 in the pharmaceutical compositions as described herein. In another embodiment, the zinc salt and peptide have a molar ratio of about 0.08:1 in the pharmaceutical compositions as described herein. In another embodiment, the zinc salt and peptide have a molar ratio of about 0.09:1 in the pharmaceutical compositions as described herein.

[0309] In some embodiments, the diluent in the pharmaceutical compositions as described herein can include an inclusion complex such as a cyclodextrin or a derivative thereof. Examples of a cyclodextrin derivative include, but are not limited to, Captisol (sulfobutyl ether β-cyclodextrin sodium salt) and Kleptose HPB (hydroxypropyl-β-cyclodextrin).

[0310] In some embodiments of the pharmaceutical compositions as described herein, the bulking agent can be a sugar alcohol, a polyol, a monosaccharide, a disaccharide, a polysaccharide, an amino acid or a polymer. Examples of sugar alcohols or polyols that can be used as a bulking agent include, but are not limited to, mannitol, sorbitol, xylitol, lactitol, isomalt, maltitol, hydrogenated starch hydrolysates (HSH) and glycerol.

[0311] Examples of monosaccharides that can be used as a bulking agent include hexoses and pentoses. Non-limiting examples of hexose bulking agents include glucose, fructose, mannose, galactose, gulose, idose, talose, allose, altrose, sorbose, tagatose and pyranose or a combination thereof. Non-limiting examples of pentose bulking agents include arabinose, lyxose, ribose, xylose, ribulose, xylulose and deoxyribose or a combination thereof.

[0312] Examples of disaccharides that can be used as a bulking agent include sucrose, lactose, maltose, trehalose, cellobiose, chitoblose, fructose, dextrose, glucose, maltose and dextran or a combination thereof. In one embodiment, the bulking agent is sucrose.

[0313] Examples of polysaccharides that can be used as a bulking agent include water soluble polysaccharides, such as pectin, pullulan, and gum Arabic.Attorney Docket No. T0947.70006WO00

[0314] Examples of amino acids that can be used as a bulking agent include any natural amino acids such as 20 naturally occurring amino acids and non-natural amino acids. In one embodiment, the bulking agent is glycine or histidine. In another embodiment, the bulking agent is glycine.

[0315] Examples of polymers that can be used as a bulking agent include polyethylene glycol, polyvinyl pyrrolidone (k12, k17) and gelatin.

[0316] In some embodiments, the bulking agent in the pharmaceutical composition as described herein is mannitol, sorbitol, xylitol or glycerol. In one embodiment, the bulking agent the bulking agent in the pharmaceutical composition as described herein is mannitol.

[0317] In some embodiments, the bulking agent in the reconstituted pharmaceutical compositions as described herein is from about 0.1% (w / v) to about 15%(w / v), about 0.5% (w / v) to about 12% (w / v), about 1% (w / v) to about 10% (w / v), about 0.5% (w / v) to about 5% (w / v), about 1% (w / v) to about 5% (w / v), about 2% (w / v) to about 10% (w / v), about 4% (w / v) to about 5% (w / v) or about 4% (w / v) to about 10% (w / v).

[0318] In some embodiments, the bulking agent in the reconstituted pharmaceutical compositions as described herein is about 1% (w / v) to about 10% (w / v).

[0319] In some embodiments, the bulking agent in the reconstituted pharmaceutical compositions as described herein is about 4% (w / v) to about 5% (w / v).

[0320] In some embodiments, the bulking agent in the reconstituted pharmaceutical compositions as described herein is about 0.1% (w / v), about 0.2% (w / v), about 0.5% (w / v), about 0.7% (w / v), about 0.8% (w / v), about 1% (w / v), about 1.5% (w / v), about 2% (w / v), about 2.5% (w / v), about 3% (w / v), about 3.5 % (w / v), about 4% (w / v), about 4.5 % (w / v), about 5% (w / v).

[0321] In one embodiment, the bulking agent in the reconstituted pharmaceutical compositions as described herein is about 2% (w / v).

[0322] In some embodiments, the bulking agent in the pharmaceutical compositions as described herein can be from about 1 mg to about 30 mg, about 10 mg to about 30 mg, or about 20 mg to about 25 mg.

[0323] In some embodiments, the bulking agent in the pharmaceutical compositions as described herein is about 1 mg, about 2 mg, about 3 mg, about 4 mg, about, 5 mg, about 6 mg, about 8 mg, about 10 mg, about 12 mg, about 15 mg, about 16 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, or about 30 mg.Attorney Docket No. T0947.70006WO00

[0324] Various buffering agents can be used in the pharmaceutical compositions as described herein. In some embodiments, the buffering agent is selected from sodium acetate, sodium acetate trihydrate, phosphates, ammonium sulfate, ammonium hydroxide, arginine, aspartic acid, benzene sulfonic acid, sodium benzoate / benzoic acid, boric acid / sodium boric acid salt, sodium carbonate, carbon dioxide, citrate, citric acid, diethanolamine, glucono delta lactone, glycine / glycine HCl, histidine / histidine HCl, hydrochloric acid, hydrobromic acid, lysine (L), maleic acid, meglumine, methanesulfonic acid, monoethanolamine, sodium hydroxide, succinate sodium / disodium, sulfuric acid, tromethamine (Tris), EDTA, tartaric acid, Bis-Tris, MOPS, MES, aspartate, ascorbate, 2,2′-[(2- amino-2-oxoethyl)azanediyl]diacetic acid (ADA), and borate.

[0325] In one embodiment, the buffering agent is sodium acetate. In another embodiment, the buffering agent is sodium acetate trihydrate.

[0326] In some embodiments, the buffering agent in the reconstituted pharmaceutical compositions as described herein is from about 0.01% (w / v) to about 1% (w / v), 0.01% (w / v) to about 0.068% (w / v), about 0.05 (w / v) to about 0.1% (w / v) or about 0.01% (w / v) to about 1% (w / v).

[0327] In some embodiments, the buffering agent in the pharmaceutical compositions as described herein is about 0.01% w / v to about 0.068% w / v.

[0328] In some embodiments, the buffering agent in the pharmaceutical compositions as described herein is about 0.01% w / v, about 0.02% w / v, about 0.03% w / v, about 0.05% w / v, about 0.06% w / v, about 0.068% w / v, about 0.07% w / v, about 0.08% w / v, about 0.09%, w / v about 0.1% w / v, about 0.2% w / v, about 0.3% w / v, about 0.4% w / v, about 0.5% w / v, about 0.6% w / v, about 0.68% w / v, about 0.7% w / v, about 0.8% w / v, about 0.9% w / v, about 1% w / v, about 1.2% w / v, about 1.3% w / v, about 1.4% w / v, about 1.5% w / v, about 1.6% w / v, about 1.7% w / v, about 1.8% w / v, about 1.9% w / v or about 2% w / v.

[0329] In some embodiments, the buffering agent in the reconstituted pharmaceutical compositions as described herein, the buffering agent is about 1 mg / mL to about 5 mg / mL or about 2 mg / mL to about 4 mg / mL.

[0330] In one embodiment, the buffering agent in the reconstituted pharmaceutical compositions as described herein is about 2.72 mg / mL.

[0331] In one embodiment, the buffering agent in the reconstituted pharmaceutical compositions as described herein is about 4.08 mg / mL.Attorney Docket No. T0947.70006WO00

[0332] The pharmaceutical compositions as described herein can further include a tonicity agent. Examples of tonicity agents include, but are not limited to, sodium chloride, dextrose, glycerin, mannitol, potassium chloride. In one embodiment, the tonicity agent is sodium chloride.

[0333] The pharmaceutical compositions as described herein can further comprise a pH adjuster, such as acetic acid or glacial acetic acid to allow the pH of the reconstituted pharmaceutical composition to be adjusted to a value suitable for injection to the patient. In one embodiment, the pH of the pharmaceutical composition is about 5.3 to about 5.6. In another embodiment, the pH of the reconstituted pharmaceutical composition is about 5.3, about 5.4, about 5.5 or about 5.6.

[0334] In one embodiment, the pH of the pharmaceutical composition is about 5.4.

[0335] The final volume of the reconstituted pharmaceutical composition suitable for injection to the subject can be from about 0.5 mL or about 1.0 mL, which can be achieved by, for example, adding a solvent such as water to the composition. In some embodiments, the volume of the reconstituted pharmaceutical compositions can be about 0.5 mL, about 0.6 mL, about 0.7 mL, about 0.8 mL, about 0.9 mL or about 1.0 mL.

[0336] In some embodiments, the reconstituted pharmaceutical composition suitable for subcutaneous injection as described herein can be formed by mixing a lyophilized pharmaceutical composition as described herein with a diluent to form a solution. The lyophilized pharmaceutical composition or lyophilized solid form comprises a peptide of formula (I) having the amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent, a buffering agent and optionally a zinc salt or a stabilizer such as a cyclodextrin or a derivative of cyclodextrin, wherein the two cysteine residues of the peptide are linked via a disulfide bond.

[0337] In some embodiments, the lyophilized pharmaceutical composition does not contain a zinc salt or a stabilizer.

[0338] In some embodiments, lyophilized pharmaceutical composition comprises a peptide of formula (I) having the amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu- Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent, a buffering agent and a zinc salt as described herein.

[0339] In some embodiments, the lyophilized pharmaceutical composition comprises a peptide of formula (I) having the amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu- Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulkingAttorney Docket No. T0947.70006WO00 agent, a buffering agent and a cyclodextrin or a derivative of cyclodextrin. In certain embodiments, the derivative of cyclodextrin is Captisol, i.e., sulfobutyl ether β-cyclodextrin sodium salt or Kleptose HPB, i.e., hydroxypropyl-β-cyclodextrin.

[0340] In some embodiments, the disclosure provides a pharmaceutical composition comprising a compound of formula I or Compound 1 suitable for injection to a patient as follows: Table 1: Composition of Compound 1 for Injection (Prior to Reconstitution) Component Quantity per Function Vialy , , , p adjustment, as needed 3A suitable quantity of Water for Injection is used in the process and is removed during the lyophilization process. Table 2: Composition of Compound 1 for Injection Diluent (0.18% Zinc Acetate) Component Weight per mL FunctionTable 3: Composition of Reconstituted Compound 1 for Injection Component Quantity per vial FunctionAttorney Docket No. T0947.70006WO00 Component Quantity per vial Function 20 mg,

[0341] In some embodiments, the disclosure provides a pharmaceutical composition comprising a compound of formula I or Compound 1 suitable for injection to a patient as follows: Table 4: Compositions of Compound 1 Injection with Zinc Chloride Composition (mg per mL) ComponentTreatmentTreatment Treatment Treatment Treatment TreatmentFunctionl nt y t [03 ] n some embod ments, t e d sc osure prov des a p armaceut ca compos t on comprising a compound of formula I or Compound 1 reconstituted for injection as follows: Table 5. Composition of reconstituted compound 1 for injection Component 10mg 15mg 20mg 30mg 45mg 60mgAttorney Docket No. T0947.70006WO00 Sodium acetate trihydrate 0.3% 0.3% 0.3% 0.3% 0.3% 0.3% Zinc acetate 0.2% 0.2% 0.2% 0.2% 0.2% 0.2%Component 10mg 15mg 20mg 30mg 45mg 60mg Compound 1 10 15 20 30 45 60Table 7. Zn:Compound 1 molar ratio of reconstituted compound 1 composition for injection Component 10mg 15mg* 20mg 30mg 45mg 60mg C m nd 1 (mM) 410 615 820 1229 1844 2459Component Quantity per Function i lSodium hydroxide (Ph Eur, BP, JP, NF) can also be used for pH adjustment, as needed 3 Range of Zn:API molar ratio is 0.2:1 to 1.2:1

[0343] In some embodiments, the disclosure provides a stable liquid pharmaceutical composition comprising a compound of formula I or Compound 1 as follows:Attorney Docket No. T0947.70006WO00 Table 9 Component % w / v Table 10Dose Strength Zn: Compound 1 (MolarTable 11A. Zn:Compound 1 = 2:1 Table 11B. Zn:Compound 1 = 2:1 Component Weight (mg / mL) Component Percentage (%)Attorney Docket No. T0947.70006WO00 Table 11C. Zn:Compound 1 = 1:1 Table 11D. Zn:Compound 1 = 1:1 Component Weight (mg / mL) Component Percentage (%) Compound 1 20 Compound 1 2Table 11E. Zn:Compound 1 = Table 11F. Zn:Compound 1 = 0.75:1 0.75:1 Component Weight (mg / mL) Component Percentage (%) Com ound 1 20 Com ound 1 2Table 12A. Zn:Compound 1 = 2:1 Table 12B. Zn:Compound 1 = 2:1 Component Weight (mg / mL) Component Percentage (%)Attorney Docket No. T0947.70006WO00 Table 12C. Zn:Compound 1 = 1:1 Table 12D. Zn:Compound 1 = 1:1 Component Weight (mg / mL) Component Percentage (%) Compound 1 45 Compound 1 4.5Table 12E. Zn:Compound 1 = Table 12F. Zn:Compound 1 = 0.75:1 0.75:1 Component Weight (mg / mL) Component Percentage (%) Compound 1 45 Compound 1 4.5

[0344] In some embodiments, the disclosure provides a stable liquid pharmaceutical formulation comprising a compound of Compound 1 suitable for injection to a patient as follows: Table 13 Component Composition (o / ow / v) PurposeAttorney Docket No. T0947.70006WO00

[0345] In some embodiments, the disclosure provides a stable liquid pharmaceutical formulation comprising a compound of Compound 1 suitable for injection to a patient as follows: Table 14 Component Range Purpose Compound 1 0.1-12% API 0068% Table 15Component Range Purpose Compound 1 0.2-4.0% API

[0346] In, q hs with less than 5% of Asp-related degradants. In certain embodiments, the amount of Asp-related degradants is less than 0.1 %, less than 0.5%, less than 1%, less than 2%, less than 3%, less than 4%, or less than 5%.

[0347] In other embodiments, the disclosure provides a lyophilized pharmaceutical composition for reconstitution comprising 10 mg of compound of formula I or Compound 1 suitable for injection to a patient. The compositions are shown in Table 16A and 16B. Table 16A Table 16B Component Percentage (%) Component Weight (mg)Attorney Docket No. T0947.70006WO00 Zinc acetate 0.2% Zinc acetate 0.9 Glacial acetic acid QS to pH5.4 Glacial acetic acid QS to pH5.4

[0348] In other embodiments, the disclosure provides a lyophilized pharmaceutical composition for reconstitution comprising 15 mg of Compound 1 (compound of formula I) suitable for injection to a patient. The composition is shown in Table 17A and 17B. Table 17A Table 17B Component Percentage (%) Component Weight (mg)Compound 1 (15 mg) 3.0% Compound 1 15closure provides a ln for reconstitution comprising 20 mg of Compound 1 (compound of formula I) suitable for injection to a patient. The composition is shown in Table 18A and 18B. Table 18A Table 18B Component Percentage (%) Component Weight (mg)

[0350] In other embodiments, the disclosure provides a lyophilized pharmaceutical composition for reconstitution comprising 30 mg of Compound 1 (compound of formula I) suitable for injection to a patient. The composition is shown in Table 19A and 19B. Table 19A Table 19B Component Percentage (%) Component Weight (mg)Attorney Docket No. T0947.70006WO00 Mannitol 4.0% Mannitol 20 Sodium acetateSodium acetatetrih drate 03%ih 1closure provides a ln for reconstitution comprising 45 mg of Compound 1 (compound of formula I) suitable for injection to a patient. The composition is shown in Table 20A and 20B. Table 20A Table 20B Component Percentage (%) Component Weight (mg)Compound 1 (45 mg) 9.0% Compound 1 45closure provides a ln for reconstitution comprising 60 mg of Compound 1 (compound of formula I) suitable for injection to a patient. The composition is shown in Table 21A and 21B. Table 21A Table 21B Component Percentage (%) Component Weight (mg)Com ound 1 (60 m ) 120% Com ound 1 60

[0353] In some embodiments, the disclosure provides a pharmaceutical composition comprising an effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average Compound 1 plasma area under the curve (average AUC) of at least 13000 ng.h / mL, at least 16000 ng.h / mL, at least 19000 ng.h / mL, at least 20000 ng.h / mL, at least 21000 ng.h / mL, at least 27000 ng.h / mL, at least 31000 ng.h / mL, at least 34000 ng.h / mL, at least 39000 ng.h / mL, at least 46000 ng.h / mL or at leastAttorney Docket No. T0947.70006WO00 64000 ng.h / mL, per each 10 mg, 20 mg, 30 mg, 45 mg or 60 mg dosage of compound of formula (I) delivered; and an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 260 ng / mL, at least 360 ng / mL, at least 400 ng / mL, at least 460 ng / mL, at least 560 ng / mL, at least 1000 ng / mL or at least 2000 ng / mL, per each 10 mg, 20 mg, 30 mg, 45 mg or 60 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration.

[0354] In certain embodiments, the disclosure provides a pharmaceutical composition comprising an effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 13000 ng.h / mL, at least 16000 ng.h / mL or at least 19000 ng.h / mL per each 10 mg dosage of compound of formula (I) delivered; an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 260 ng / mL or at least 320 ng / mL per each 10 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 10 mg dosage, the AUC is at least 16000 ng.h / mL and Cmax is at least 260 ng / mL.

[0355] In certain embodiments, the disclosure provides a pharmaceutical composition comprising an effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 16000 ng.h / mL, at least 21000 ng.h / mL or at least 26000 ng.h / mL per each 20 mg dosage of Compound 1 delivered; an average maximum compound of formula (I) blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 260 ng / mL or at least 340 ng / mL per each 20 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 20 mg dosage, the AUC is at least 21000 ng.h / mL and Cmax is at least 260 ng / mL.

[0356] In certain embodiments, the disclosure provides a pharmaceutical composition comprising an effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 20000 ng.h / mL, at least 27000 ng.h / mL or at least 34000 ng.h / mL per each 30 mg dosage of compound of formula (I) delivered; an averageAttorney Docket No. T0947.70006WO00 maximum Compound 1 blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 300 ng / mL or at least 400 ng / mL per each 30 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 30 mg dosage, the AUC is at least 27000 ng.h / mL and Cmaxis at least 300 ng / mL.

[0357] In certain embodiments, the disclosure provides a pharmaceutical composition comprising an effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 31000 ng.h / mL, at least 39000 ng.h / mL or at least 47000 ng.h / mL per each 45 mg dosage of compound of formula (I) delivered; an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 360 ng / mL, at least 460 ng / mL or at least 560 ng / mL per each 45 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 45 mg dosage, the AUC is at least 39000 ng.h / mL and Cmaxis at least 460 ng / mL.

[0358] In certain embodiments, the disclosure provides a pharmaceutical composition comprising an effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 28000 ng.h / mL, at least 46000 ng.h / mL or at least 64000 ng.h / mL per each 60 mg dosage of compound of formula (I) delivered; an average maximum compound of formula (I) blood plasma concentration (average Cmax) of at least 1000 ng / mL or at least 1000 ng / mL per each 60 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 60 mg dosage, the AUC is at least 46000 ng.h / mL and Cmaxis at least 1000 ng / mL.

[0359] In another aspect, the disclosure provides a method for preparing a lyophilized pharmaceutical composition, the method comprising: preparing a solution comprising a peptide of formula (I) having the amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu- Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent, a buffering agent, a solvent and optionally a zinc salt to a suitable pH, wherein the two cysteine residues of the peptide are linked via a disulfide bond; and drying the solution byAttorney Docket No. T0947.70006WO00 lyophilization to afford a lyophilized pharmaceutical composition. Thus, the peptide is cyclized through a disulfide bond formed by the two cysteine residues in the peptide. In certain embodiments, the solution comprises a peptide having the amino acid sequence: Isovaleric acid- DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent as described herein, a solvent and a buffering agent as described herein. In other embodiments, the solution comprises a peptide having the amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt thereof, a bulking agent as described herein, a buffering agent as described herein, a solvent and a zinc salt as described herein. In some embodiments, the solvent is water, DMSO, DMF or a combination thereof. In some embodiments, the solution is an aqueous solution. In some embodiments, the solution is prepared by dissolving the peptide of formula (I) or a pharmaceutically acceptable salt, a bulking agent, a buffering agent and optionally a zinc salt in water. In one embodiment, the lyophilized pharmaceutical composition is a powder. In another embodiment, the suitable pH is about 5.4.

[0360] In another aspect, the disclosure provides a method for preparing a pharmaceutical composition suitable for subcutaneous administration to a subject, the method comprising mixing a lyophilized pharmaceutical composition with a diluent to obtain a solution suitable for subcutaneous administration, wherein the lyophilized pharmaceutical composition comprises a peptide of formula (I) having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt, a bulking agent as described herein, a buffering agent as described herein and optionally a first zinc salt; and wherein the diluent comprises optionally a second zinc salt as and a solvent; and wherein the two cysteine residues of the peptide are linked to form a disulfide bond. In some embodiments, the first zinc salt and the second zinc salt can be the same zinc salt as described herein. In other embodiments, the first zinc salt and the second zinc salt can be different zinc salts as described herein.

[0361] In some embodiments of the method for preparing a pharmaceutical composition suitable for subcutaneous administration to a subject, the method comprises mixing a lyophilized pharmaceutical composition with a diluent to obtain a solution suitable for subcutaneous administration, wherein the lyophilized pharmaceutical composition comprises a peptide having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt, a bulking agent and a buffering agent; wherein the diluentAttorney Docket No. T0947.70006WO00 comprises a zinc salt as described herein and a solvent; and wherein the two cysteine residues in the peptide are linked to form a disulfide bond.

[0362] In some embodiments of the method for preparing a pharmaceutical composition suitable for subcutaneous administration to a subject, the method comprises mixing a lyophilized pharmaceutical composition with a diluent to obtain a solution suitable for subcutaneous administration, wherein the lyophilized pharmaceutical composition comprises a peptide having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt, a bulking agent, a buffering agent and a first zinc salt; wherein the diluent comprises a second zinc salt and a solvent; and wherein the two cysteine residues in the peptide are linked to form a disulfide bond. In one embodiment, the first zinc salt and the second zinc salt can be the same zinc salt as described herein. In another embodiment, the first zinc salt and the second zinc salt can be different zinc salts as described herein.

[0363] In some embodiments of the method for preparing a pharmaceutical composition suitable for subcutaneous administration to a subject, the method comprises mixing a lyophilized pharmaceutical composition and a diluent to obtain a solution suitable for subcutaneous administration, wherein the lyophilized pharmaceutical composition comprises a peptide having the amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2(SEQ ID NO: 1) or a pharmaceutically acceptable salt, a bulking agent as described herein, a buffering agent as described herein and a zinc salt as described herein; wherein the diluent comprises a solvent, but not a zinc salt; and wherein the two cysteine residues of the peptide are linked to form a disulfide bond.

[0364] In another aspect, the disclosure provides metabolites of Compound 1 that provide biological activity as described and accordingly the compounds of the disclosure includes pharmaceutical compositions thereof and methods of use described herein are equally applicable to the metabolites, particularly metabolites, M1, M4, and M9. Thus, in certain aspects, a pharmaceutical composition can comprise synthesized peptides comprising the structures of M1, M4, M6, and / or M9.

[0365] In certain embodiments, a metabolite of Compound 1 has a sequence comprising: Isovaleric_Acid-D-T-H-F-P-Cys_S-K-G-C-Kam-I-Lys_IsoGlu_Palm-F-E-P-R SEQ ID NO: 2 (also referred to as M1) or a pharmaceutically acceptable salt or hydrate thereof. In one aspect, a pharmaceutical composition comprises a synthesized peptide having the structure of M1, wherein synthesized peptide M1 is present in the composition in a greater amount than any other active ingredient. In another aspect, a pharmaceutical composition comprises at least 10, 20, 30, 40, 50, 60,Attorney Docket No. T0947.70006WO00 70.80, 90, 95 or 100% of M1 of the total peptide content of the pharmaceutical composition. In another aspect, M1 has an amine group at the C-terminus. In another aspect, the isovaleric acid of M1 is substituted with Ra-C(O)- or hydrogen, wherein Ra is C1-20 alkyl or C3-8 cycloalkyl or absent.

[0366] In certain embodiments, the metabolite M1 comprises the following structure:Attorney Docket No. T0947.70006WO00

[0367] In certain embodiments, a metabolite of Compound 1 has a sequence comprising: Isovaleric_Acid-D-T-H-F-P-Cys_Cys-I-Lys_IsoGlu_Palm-F-E-P-R SEQ ID NO: 3 (also referred to as M4) or a pharmaceutically acceptable salt or hydrate thereof. In one aspect, a pharmaceutical composition comprises a synthesized peptide having the structure of M4, wherein synthesized peptide M4 is present in the composition in a greater amount than any other active ingredient.

[0368] In certain embodiments, the metabolite M4 comprises the following structure:Attorney Docket No. T0947.70006WO00

[0369] In certain embodiments, a metabolite of Compound 1 has a sequence comprising: Isovaleric_Acid-D-T-H-F-P-C-I-Lys_IsoGlu_14OHPalm-F-E-P-R-S-K-G-C-K-am, wherein the thiol side chains of the cysteines form an intramolecular disulfide bond (i.e., cyclized) SEQ ID NO: 4 (also referred to as M9) or a pharmaceutically acceptable salt or hydrate thereof. In one aspect, a pharmaceutical composition comprises a synthesized peptide having the structure of M9, wherein synthesized peptide M9 is present in the composition in a greater amount than any other active ingredient.

[0370] In certain embodiments, the metabolite M9 comprises the following structure:

[0371] In certain embodiments, a metabolite of Compound 1 has a sequence comprising: Lys_IsoGlu_Palm SEQ ID NO: 5 (also referred to as M6) or a pharmaceutically acceptable salt or hydrate thereof. In one aspect, a pharmaceutical composition comprises a synthesized peptideAttorney Docket No. T0947.70006WO00 having the structure of M6, wherein synthesized peptide M6 is present in the composition in a greater amount than any other active ingredient.

[0372] In certain embodiments, the metabolite M6 comprises the following structure:

[0373] Metabolites M1 and M6 were detected sporadically in plasma in various subjects. Across all dose levels, Tmax for M9 and M4 appeared later than for Compound 1. M9 exposure was approximately 25-30% of total drug-related exposure (AUC) across the tested dose levels. M4 exposure was approximately 20% of total drug-related exposure in the PFS formulation. For the lyophilized product, the percentage of M4 exposure relative to total drug-related exposure increased with dose, ranging from 1.9 % at 10 mg to 18.5% at 60 mg. Therefore, M9 is considered a major metabolite (≥10%) and conservatively, M4 is also considered a major metabolite for this single dose study. Given the low levels of M1 and M6 (<1%), these two metabolites are considered minor metabolites.Attorney Docket No. T0947.70006WO00 Table 22A Metabolite (Compound 1) / Total Drug Ratio (%) M1 M4 M6 M9 Compound 1Table 22B Molecular IC50 Relative

[0374] In another aspec , presen sc osure prov es me o s or rea ng a subject afflicted with a disease or disorder associated with dysregulated hepcidin signaling, wherein the method comprises administering to the subject a pharmaceutical composition as disclosed herein. In one embodiment, a method is provided for treating a subject afflicted with a disease or disorder characterized by increased activity or expression of ferroportin, wherein the method comprises administering to the subject a pharmaceutical composition as disclosed herein in an amount sufficient to (partially or fully) bind to and agonize ferroportin in the subject. In one embodiment, a method is provided for treating a subject afflicted with a disease or disorder characterized by dysregulated iron metabolism,Attorney Docket No. T0947.70006WO00 wherein the method comprises administering to the subject a pharmaceutical composition as disclosed herein.

[0375] In some embodiments, the subject in need thereof has been diagnosed with or has been determined to be at risk of developing a disease or disorder characterized by dysregulated iron levels (e.g., diseases or disorders of iron metabolism; diseases or disorders related to iron overload; and diseases or disorders related to abnormal hepcidin activity or expression). In certain embodiments, the subject is a mammal (e.g., a human).

[0376] In another aspect, the disclosure provides a method for treating a disease of iron metabolism in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein.

[0377] In some embodiments, the disclosure provides a method for treating polycythemia vera in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein. In some embodiments, the composition is administered to the subject subcutaneously.

[0378] The diseases or disorders that are treatable or preventable with the pharmaceutical compositions as described herein include a disease of iron metabolism, such as, e.g., an iron overload disease, iron deficiency disorder, disorder of iron biodistribution, or another disorder of iron metabolism and other disorder potentially related to iron metabolism, etc. In particular embodiments, the disease of iron metabolism is hemochromatosis, HFE mutation hemochromatosis, ferroportin mutation hemochromatosis, transferrin receptor 2 mutation hemochromatosis, hemojuvelin mutation hemochromatosis, hepcidin mutation hemochromatosis, juvenile hemochromatosis, neonatal hemochromatosis, hepcidin deficiency, transfusional iron overload, thalassemia, thalassemia intermedia, alpha thalassemia, beta thalassemia, sideroblastic anemia, porphyria, porphyria cutanea tarda, African iron overload, hyperferritinemia, ceruloplasmin deficiency, atransferrinemia, congenital dyserythropoietic anemia, anemia of chronic disease, anemia of inflammation, anemia of infection, hypochromic microcytic anemia, iron- deficiency anemia, iron-refractory iron deficiency anemia, anemia of chronic kidney disease, transfusion- dependent anemia, hemolytic anemia, erythropoietin resistance, iron deficiency of obesity, other anemias, benign or malignant tumors that overproduce hepcidin or induce its overproduction, conditions with hepcidin excess, Friedreich ataxia, gracile syndrome, Hallervorden-Spatz disease, Wilson's disease, pulmonary hemosiderosis, hepatocellular carcinoma, cancer (e.g., liver cancer),Attorney Docket No. T0947.70006WO00 hepatitis, cirrhosis of liver, pica, chronic renal failure, insulin resistance, diabetes, atherosclerosis, neurodegenerative disorders, dementia, multiple sclerosis, Parkinson's disease, Huntington's disease, or Alzheimer's disease.

[0379] In certain embodiments, the diseases or disorders treatable with the pharmaceutical compositions as described herein are related to iron overload diseases such as iron hemochromatosis, HFE mutation hemochromatosis, ferroportin mutation hemochromatosis, transferrin receptor 2 mutation hemochromatosis, hemojuvelin mutation hemochromatosis, hepcidin mutation hemochromatosis, juvenile hemochromatosis, neonatal hemochromatosis, hepcidin deficiency, transfusional iron overload, thalassemia, thalassemia intermedia, alpha thalassemia.

[0380] In certain embodiments, the diseases, or disorders treatable with the compositions as described herein include diabetes (Type I or Type II), insulin resistance, glucose intolerance and other disorders may be ameliorated by treating underlying iron metabolism disorders.

[0381] In certain embodiments, the disease or disorder treatable with the liquid compositions as described herein includes obesity associated with a high calorie diet, diet-induced obesity, hepatic steatosis (fatty liver disease), hepatic stenosis, hepatic fibrosis, metabolic dysfunction-associated fatty liver disease (MAFLD), porphyria, cholangitis, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis (NASH) and alcohol fatty liver disease.

[0382] In certain embodiments, the disease or disorder is postmenopausal osteoporosis.

[0383] In some embodiments, the disease of iron metabolism is an iron overload disease.

[0384] In certain embodiments, the diseases of iron metabolism are iron overload diseases, which include hereditary hemochromatosis, iron-loading anemias, alcoholic liver diseases, heart disease and / or failure, cardiomyopathy, and chronic hepatitis C.

[0385] In certain embodiments, any of these diseases, disorders, or indications are caused by or associated with a deficiency of hepcidin or iron overload.

[0386] In some embodiments, the disclosure provides a method for treating polycythemia vera in a subject such as a human patient in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein.

[0387] In some embodiments, the method of treating polycythemia vera in a subject, for example, a human patient, comprises subcutaneously administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject at such a rate produces in the subject:Attorney Docket No. T0947.70006WO00 an average Compound 1 plasma area under the curve (average AUC) of at least 13000 ng.h / mL, at least 16000 ng.h / mL, at least 19000 ng.h / mL, at least 20000 ng.h / mL, at least 21000 ng.h / mL, at least 27000 ng.h / mL, at least 31000 ng.h / mL, at least 34000 ng.h / mL, at least 39000 ng.h / mL, at least 46000 ng.h / mL or at least 64000 ng.h / mL, per each 10 mg, 20 mg, 30 mg, 45 mg or 60 mg dosage of compound of formula (I) delivered; and an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 260 ng / mL, at least 360 ng / mL, at least 400 ng / mL, at least 460 ng / mL, at least 560 ng / mL, at least 1000 ng / mL or at least 2000 ng / mL, per each 10 mg, 20 mg, 30 mg, 45 mg or 60 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration.

[0388] In certain embodiments, the method of treating polycythemia vera in a subject, for example, a human patient comprises subcutaneously administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject at such a rate produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 13000 ng.h / mL, at least 16000 ng.h / mL or at least 19000 ng.h / mL per each 10 mg dosage of compound of formula (I) delivered; an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 260 ng / mL or at least 320 ng / mL per each 10 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 10 mg dosage, the AUC is at least 16000 ng.h / mL and Cmaxis at least 260 ng / mL.

[0389] In certain embodiments, the method of treating polycythemia vera in a subject, for example, a human patient comprises subcutaneously administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 16000 ng.h / mL, at least 21000 ng.h / mL or at least 26000 ng.h / mL per each 20 mg dosage of Compound 1 delivered; an average maximum compound of formula (I) blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 260 ng / mL or at least 340 ng / mL per each 20 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 20 mg dosage, the AUC is at least 21000 ng.h / mL and Cmaxis at least 260 ng / mL.Attorney Docket No. T0947.70006WO00

[0390] In certain embodiments, the method of treating polycythemia vera in a subject, for example, a human patient comprises subcutaneously administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 20000 ng.h / mL, at least 27000 ng.h / mL or at least 34000 ng.h / mL per each 30 mg dosage of compound of formula (I) delivered; an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 200 ng / mL, at least 300 ng / mL or at least 400 ng / mL per each 30 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 30 mg dosage, the AUC is at least 27000 ng.h / mL and Cmaxis at least 300 ng / mL.

[0391] In certain embodiments, the method of treating polycythemia vera in a subject, for example, a human patient comprises subcutaneously administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 31000 ng.h / mL, at least 39000 ng.h / mL or at least 47000 ng.h / mL per each 45 mg dosage of compound of formula (I) delivered; an average maximum Compound 1 blood plasma concentration (average Cmax) of at least 360 ng / mL, at least 460 ng / mL or at least 560 ng / mL per each 45 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurable concentration. In one embodiment, with a 45 mg dosage, the AUC is at least 39000 ng.h / mL and Cmax is at least 460 ng / mL.

[0392] In certain embodiments, the method of treating polycythemia vera in a subject, for example, a human patient comprises subcutaneously administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of peptide of formula (I), wherein subcutaneous administration of the pharmaceutical composition to a subject produces in the subject: an average compound of formula (I) plasma area under the curve (average AUC) of at least 28000 ng.h / mL, at least 46000 ng.h / mL or at least 64000 ng.h / mL per each 60 mg dosage of compound of formula (I) delivered; an average maximum compound of formula (I) blood plasma concentration (average Cmax) of at least 1000 ng / mL or at least 1000 ng / mL per each 60 mg dosage of compound of formula (I) delivered; wherein the AUC is measured from time zero to the time of last measurableAttorney Docket No. T0947.70006WO00 concentration. In one embodiment, with a 60 mg dosage, the AUC is at least 46000 ng.h / mL and Cmax is at least 1000 ng / mL.

[0393] In another aspect, the disclosure provides a pharmaceutical composition comprising a peptide of formula (I) to a subject in need thereof in combination with a second therapeutic agent. In certain embodiments, the second therapeutic agent is provided to the subject before and / or simultaneously with and / or after the pharmaceutical composition is administered to the subject. In one embodiment, the second therapeutic agent is iron chelator. In certain embodiments, the second therapeutic agent is selected from the iron chelators Deferoxamine and Deferasirox (Exjade ™). In another embodiment, the method comprises administering to the subject a third therapeutic agent. Administration Methods

[0394] The pharmaceutical composition as described herein containing peptide of formula (I) can be administered to the subject, for example, a human patient at various frequency, such as an interval around the clock, hourly, every four hours, once daily, twice daily, three times daily, four times daily, every other day, once weekly, twice weekly, once bi-weekly, twice bi-weekly, once monthly, twice monthly, or three times per month.

[0395] In some embodiments, the pharmaceutical composition as described herein can be administered at a dose from about 1 mg / day to about 500 mg / day. In other embodiments, the pharmaceutical composition as described herein can be administered at a dose from about 1 mg / m2to about 3 g / m2, from about 5 mg / m2to about 1 g / m2, or from about 10 mg / m2to about 500 mg / m2.

[0396] The administered dose may be expressed in units of mg / m2 / day in which a patient’s body surface area (BSA) may be calculated in m2using various available formulae using the patient’s height and weight. The administered dose may alternatively be expressed in units of mg / day which does not take into consideration the patient’s BSA. It is straightforward to convert from one unit to another given a patient’s height and weight.

[0397] In one embodiment, the treatment is carried out for one or more treatment cycles. By “treatment cycle”, it is meant a pre-determined period of time for administering the pharmaceutical composition as described herein and optionally at least one combination partner. Typically, the patient is examined at the end of each treatment cycle to evaluate the effect of the therapy. In one embodiment, the administration is carried out for 1 to 48 treatment cycles. In another embodiment, co-administration is carried out for 1 to 36 treatment cycles. In another embodiment, the administration is carried out for 1 to 24 treatment cycles.Attorney Docket No. T0947.70006WO00

[0398] In one embodiment, each of the treatment cycle has about 3 or more days. In another embodiment, each treatment cycle lasts from about 3 days to about 60 days. In another embodiment, each treatment cycle has from about 5 days to about 50 days. In another embodiment, each of the treatment cycle has from about 7 days to about 28 days. In another embodiment, each of the treatment cycle has 28 days. In one embodiment, the treatment cycle has about 29 days. In another embodiment, the treatment cycle has about 30 days. In another embodiment, the treatment cycle has about a month-long treatment cycle. In another embodiment, the treatment cycle lasts from about 4 to about 6 weeks.

[0399] Depending on the patient’s condition and the intended therapeutic effect, the dosing frequency for the peptide of formula (I), or a pharmaceutically acceptable salt, solvate, thereof and may vary from once per day to six times per day. That is, the dosing frequency may be once per day, twice per day, three times per day, four times per day, five times per day, or six times per day. In some embodiments, dosing frequency may be one to six times per week or one to four times per month. In one embodiment, dosing frequency may be once a week, once every two weeks, once every three weeks, once every four weeks, or once a month.

[0400] There may be one or more void days in a treatment cycle. By “void day”, it is meant a day when neither the peptide of formula (I), or a pharmaceutically acceptable salt, solvate thereof or at least one combination partner is administered. In other words, none of the peptide of formula (I), or a pharmaceutically acceptable salt or solvate thereof and / or at least one combination partner is administered on a void day. Any treatment cycle must have at least one non-void day. By “non-void day”, it is meant a day when a peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof and optionally at least one combination partner is administered.

[0401] By “simultaneous administration”, it is meant that the peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof and at least one combination partner are administered on the same day. For the simultaneous administration, the peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof and optionally at least one combination partner can be administered at the same time or one at a time.

[0402] The term “co-administration” or “coadministration” refers to administration of a peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and (b) at least one combination partner, together in a coordinated fashion. For example, co- administration can be simultaneous administration, sequential administration, overlappingAttorney Docket No. T0947.70006WO00 administration, interval administration, continuous administration, or a combination thereof. In one embodiment, the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and at least one combination partner, also for treatment of integrin-related disorders, are formulated into a single dosage form. In another embodiment, the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and at least one combination partner are provided in a separate dosage forms.

[0403] In one embodiment, the peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof, is administered from 1 to 4 times per day, 1 to 4 times per week, once every two weeks, once every three weeks, once every four weeks or 1 to 4 times per month. In another embodiment, the peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof, is administered once a week, once every two weeks, once every three weeks, once every four weeks, or once a month.

[0404] By “sequential administration”, it is meant that during a period of two or more days of continuous co-administration without any void day, only a peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof and at least one combination partner is administered on any given day.

[0405] By “overlapping administration”, it is meant that during a period of two or more days of continuous co-administration without any void day, there is at least one day of simultaneous administration and at least one day when only a peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof and at least one combination partner is administered.

[0406] By “interval administration”, it is meant a period of co-administration with at least one void day. By “continuous administration”, it is meant a period of co-administration without any void day. The continuous administration may be simultaneous, sequential, or overlapping, as described above.

[0407] In some embodiments, the present disclosure provides a pharmaceutical composition and / or combination comprising a peptide of formula (I), or a pharmaceutically acceptable salt, or solvate thereof, as disclosed herein, combined with a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known to those skilled in the art and include, but are not limited to, from about 0.01 to about 0.1 M and preferably 0.05M phosphate buffer or 0.8% saline. Such pharmaceutically acceptable carriers can be aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents suitable for use in the present application include, butAttorney Docket No. T0947.70006WO00 are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.

[0408] Aqueous carriers suitable for use in the present application include, but are not limited to, water, ethanol, alcoholic / aqueous solutions, glycerol, emulsions or suspensions, including saline and buffered media.

[0409] Liquid carriers suitable for use in the present application can be used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compounds. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmo-regulators.

[0410] Liquid carriers suitable for use in the present application include, but are not limited to, water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, and oils (e.g. fractionated coconut oil and arachis oil). For parenteral administration, the carrier can also include an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are useful in sterile liquid form comprising compounds for parenteral administration. The liquid carrier for pressurized compounds disclosed herein can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.

[0411] Solid carriers suitable for use in the present application include, but are not limited to, inert substances such as lactose, starch, glucose, methyl-cellulose, magnesium stearate, dicalcium phosphate, mannitol and the like. A solid carrier can further include one or more substances acting as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material. In powders, the carrier can be a finely divided solid which is in admixture with the finely divided active compound. In tablets, the active compound is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active compound. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins. A tablet may be made byAttorney Docket No. T0947.70006WO00 compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free flowing form such as a powder or granules, optionally mixed with a binder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cellulose) surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropyl methylcellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

[0412] Parenteral carriers suitable for use in the present application include, but are not limited to, sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's and fixed oils. Intravenous carriers include fluid and nutrient replenishers, electrolyte replenishers such as those based on Ringer's dextrose and the like. Preservatives and other additives can also be present, such as, for example, antimicrobials, antioxidants, chelating agents, inert gases and the like.

[0413] Carriers suitable for use in the present application can be mixed as needed with disintegrants, diluents, granulating agents, lubricants, binders and the like using conventional techniques known in the art. The carriers can also be sterilized using methods that do not deleteriously react with the compounds, as is generally known in the art.

[0414] Diluents may be added to the formulations of the present disclosure. Diluents increase the bulk of a solid pharmaceutical composition and / or combination and may make a pharmaceutical dosage form containing the composition and / or combination easier for the patient and care giver to handle. Diluents for solid compositions and / or combinations include, for example, microcrystalline cellulose (e.g., AVICEL), microfine cellulose, lactose, starch, pregelatinized starch, calcium carbonate, calcium sulfate, sugar, dextrates, dextrin, dextrose, dibasic calcium phosphate dihydrate, tribasic calcium phosphate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylates (e.g., EUDRAGIT(r)), potassium chloride, powdered cellulose, sodium chloride, sorbitol, and talc.

[0415] Flavoring agents and flavor enhancers make the dosage form more palatable to the patient. Common flavoring agents and flavor enhancers for pharmaceutical products that may be included inAttorney Docket No. T0947.70006WO00 the composition and / or combination of the present disclosure include maltol, vanillin, ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol, and tartaric acid.

[0416] Solid and liquid compositions and / or combinations may also be dyed using any pharmaceutically acceptable colorant to improve their appearance and / or facilitate patient identification of the product and unit dosage level.

[0417] In liquid pharmaceutical compositions and / or combinations may be prepared using the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof, of the present disclosure and any other solid excipients where the components are dissolved or suspended in a liquid carrier such as water, vegetable oil, alcohol, polyethylene glycol, propylene glycol, or glycerin.

[0418] For example, formulations for parenteral administration can contain as common excipients sterile water or saline, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, hydrogenated naphthalenes and the like. In particular, biocompatible, biodegradable lactide polymer, lactide / glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers can be useful excipients to control the release of active compounds. Other potentially useful parenteral delivery systems include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes.

[0419] Liquid pharmaceutical compositions and / or combinations may contain emulsifying agents to disperse uniformly throughout the composition and / or combination an active ingredient or other excipient that is not soluble in the liquid carrier. Emulsifying agents that may be useful in liquid compositions and / or combinations of the present disclosure include, for example, gelatin, egg yolk, casein, cholesterol, acacia, tragacanth, chondrus, pectin, methyl cellulose, carbomer, cetostearyl alcohol, and cetyl alcohol.

[0420] Liquid pharmaceutical compositions and / or combinations may also contain a viscosity enhancing agent to improve the mouth-feel of the product and / or coat the lining of the gastrointestinal tract. Such agents include acacia, alginic acid bentonite, carbomer, carboxymethylcellulose calcium or sodium, cetostearyl alcohol, methyl cellulose, ethylcellulose, gelatin guar gum, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, maltodextrin, polyvinyl alcohol, povidone, propylene carbonate, propylene glycol alginate, sodium alginate, sodium starch glycolate, starch tragacanth, and xanthan gum.Attorney Docket No. T0947.70006WO00

[0421] Sweetening agents such as aspartame, lactose, sorbitol, saccharin, sodium saccharin, sucrose, aspartame, fructose, mannitol, and invert sugar may be added to improve the taste.

[0422] Preservatives and chelating agents such as alcohol, sodium benzoate, butylated hydroxyl toluene, butylated hydroxyanisole, and ethylenediamine tetraacetic acid may be added at levels safe for ingestion to improve storage stability.

[0423] A liquid composition and / or combination may also contain a buffer such as gluconic acid, lactic acid, citric acid or acetic acid, sodium gluconate, sodium lactate, sodium citrate, or sodium acetate (sodium acetate trihydrate). Selection of excipients and the amounts used may be readily determined by the formulation scientist based upon experience and consideration of standard procedures and reference works in the field.

[0424] The lyophilized solid compositions and / or combination of the present disclosure include powders, granulates, aggregates and compacted compositions and / or combinations.

[0425] The pharmaceutical compositions and / or pharmaceutical combinations of the disclosure may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butane-diol or prepared as a lyophilized powder. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables. Formulations for intravenous administration can comprise solutions in sterile isotonic aqueous buffer. Where necessary, the formulations can also include a solubilizing agent and a local anesthetic to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampule or sachet indicating the quantity of active agent. Where the compound is to be administered by infusion, it can be dispensed in a formulation with an infusion bottle containing sterile pharmaceutical grade water, saline or dextrose / water. Where the compound is administered by injection, an ampule of sterileAttorney Docket No. T0947.70006WO00 water for injection or saline can be provided so that the ingredients can be mixed prior to administration.

[0426] Suitable formulations further include aqueous and non-aqueous sterile injection solutions that can contain antioxidants, buffers, bacteriostats, bactericidal antibiotics and solutes that render the formulation isotonic with the bodily fluids of the intended recipient; and aqueous and non-aqueous sterile suspensions, which can include suspending agents and thickening agents.

[0427] In one embodiment, a dosage form may be provided as a kit comprising the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and diluents as separate components. In one embodiment, a dosage form may be provided as a kit comprising the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof and pharmaceutically acceptable excipients and carriers as separate components. In one embodiment, a dosage form may be provided as a kit comprising the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof, at least one combination partner, and pharmaceutically acceptable excipients and carriers as separate components. In some embodiments, the dosage form kit allow physicians and patients to formulate an oral solution or injection solution prior to use by dissolving, suspending, or mixing the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof with pharmaceutically acceptable excipients and carriers. In one embodiment, a dosage form kit which provides the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof which has improved stability when compared to pre-formulated formulations of the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof.

[0428] In one embodiment, the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof is used in the formulation. The peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof, of the present disclosure may be used in pharmaceutical formulations or compositions and / or combinations as single components or mixtures together with other forms of the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof or at least one of the combination partners described herein. In one embodiment, pharmaceutical formulations or compositions and / or combinations of the present disclosure contain 25-100% or 50- 100% by weight of at least a peptide compound, or a pharmaceutically acceptable salt, solvate,Attorney Docket No. T0947.70006WO00 stereoisomer, ester, tautomer or prodrug thereof, as described herein, in the formulation or composition and / or combination and pharmaceutically acceptable excipients and carriers as separate components. In one embodiment, a dosage form may be provided as a kit comprising the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof, at least one combination partner, and pharmaceutically acceptable excipients and carriers as separate components. In some embodiments, the dosage form kit allow physicians and patients to formulate an oral solution or injection solution prior to use by dissolving, suspending, or mixing the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof with pharmaceutically acceptable excipients and carriers. In one embodiment, a dosage form kit which provides the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof which has improved stability when compared to pre-formulated formulations comprising the peptide compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, ester, tautomer or prodrug thereof.

[0429] In one embodiment, the compounds or the combination of compounds of the present disclosure can be administered in a form suitable for immediate release, delayed release, extended release, or sustained release. Immediate release, delayed release, extended release, or sustained release can be achieved by the use of suitable pharmaceutical compositions and / or pharmaceutical combinations comprising the compounds presented in this disclosure, for example, by using polymer coatings using enteric polymers, water-soluble polymers, water-insoluble polymers, gastrosoluble polymers, and combinations thereof, which are commonly known in the art. EXAMPLES Example 1: Synthesis of Compound 1

[0430] Compound 1 (also known as peptide of formula (I) or Compound of formula (I)) has SEQ ID NO: 1: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2, (SEQ ID NO: 1) , wherein two cysteine residues form a disulfide bond. One of skill in the art will appreciate that standard methods of peptide synthesis may be used to generate Compound 1. Procedure for solid-phase synthesis of peptides

[0431] Compound 1 was chemically synthesized using optimized 9-fluorenylmethoxy carbonyl (Fmoc) solid phase peptide synthesis protocols. For C-terminal amides, rink-amide resin was used, although wang and trityl resins were also used to produce C-terminal acids. The side chainAttorney Docket No. T0947.70006WO00 protecting groups were as follows: Glu, Thr and Tyr: O-tButyl; Trp and Lys: t-Boc (t- butyloxycarbonyl); Arg: N-gamma-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl; His, Gln, Asn, Cys: Trityl. For selective disulfide bridge formation, Acm (acetamidomethyl) was also used as a Cys protecting group. For coupling, a four to ten-fold excess of a solution containing Fmoc amino acid, HBTU and DIPEA (1:1:1.1) in DMF was added to swelled resin [HBTU: O-(Benzotriazol-1- yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate; DIPEA: diisopropylethylamine; DMF: dimethylformamide]. HATU (O-(7-azabenzotriazol-1-yl)-1,1,3,3,-tetramethyluronium hexafluorophosphate) was used instead of HBTU to improve coupling efficiency in difficult regions. Fmoc protecting group removal was achieved by treatment with a DMF, piperidine (2:1) solution. Procedure for cleavage of peptides off resin

[0432] Side chain deprotection and cleavage of the peptide was achieved by stirring dry resin in a solution containing trifluoroacetic acid, water, ethanedithiol and tri-isopropylsilane (90:5:2.5:2.5) for 2 to 4 hours. Following TFA removal, peptide was precipitated using ice-cold diethyl ether. The solution was centrifuged and the ether was decanted, followed by a second diethyl ether wash. The peptide was dissolved in an acetonitrile, water solution (1:1) containing 0.1% TFA (trifluoroacetic acid) and the resulting solution was filtered. The linear peptide quality was assessed using electrospray ionization mass spectrometry (ESI-MS). Procedure for purification of Peptides

[0433] Purification of the peptide compound 1 was achieved using reverse-phase high performance liquid chromatography (RP-HPLC). Analysis was performed using a C18 column (3µm, 50 x 2mm) with a flow rate of 1 mL / min. Purification of the linear peptides was achieved using preparative RP- HPLC with a C18 column (5µm, 250 x 21.2 mm) with a flow rate of 20 mL / min. Separation was achieved using linear gradients of buffer B in A (Buffer A: Aqueous 0.05% TFA; Buffer B: 0.043% TFA, 90% acetonitrile in water). Procedure for oxidation of peptides

[0434] Method A (Single disulfide oxidation). Oxidation of the unprotected peptide compound 1 was achieved by adding drop-wise iodine in MeOH (1 mg per 1 mL) to the peptide in a solution (ACN: H2O, 7: 3, 0.5% TFA). After stirring for 2 min, ascorbic acid portion wise was added until the solution was clear and the sample was immediately loaded onto the HPLC for purificationAttorney Docket No. T0947.70006WO00

[0435] Method B (Selective oxidation of two disulfides). When more than one disulfide was present (e.g., Compound 30), selective oxidation was often performed. Oxidation of the free cysteines was achieved at pH 7.6 NH4CO3 solution at 1mg / 10 mL of peptide. After 24 h stirring and prior to purification the solution was acidified to pH 3 with TFA followed by lyophilization. The resulting single oxidized peptides (with ACM protected cysteines) were then oxidized / selective deprotection using iodine solution. The peptide (1 mg per 2 mL) was dissolved in MeOH / H20, 80:20 iodine dissolved in the reaction solvent was added to the reaction (final concentration: 5 mg / mL) at room temperature. The solution was stirred for 7 minutes before ascorbic acid was added portion wise until the solution is clear. The solution was then loaded directly onto the HPLC.

[0436] Method C (Native oxidation). When more than one disulfide was present and when not performing selective oxidations, native oxidation was performed. Native oxidation was achieved with 100 mM NH4CO3 (pH7.4) solution in the presence of oxidized and reduced glutathione (peptide / GSH / GSSG, 1:100:10 molar ratio) of (peptide: GSSG: GSH, 1:10, 100). After 24 h stirring and prior to RP-HPLC purification the solution was acidified to pH 3 with TFA followed by lyophilization.

[0437] Procedure of Cysteine oxidation to produce dimers. Oxidation of compound 1 was achieved by adding drop-wise iodine in MeOH (1 mg per 1 mL) to the peptide in a solution (ACN: H2O, 7: 3, 0.5% TFA). After stirring for 2 min, ascorbic acid portion wise was added until the solution was clear and the sample was immediately loaded onto the HPLC for purification. Example 2: Preparation of lyophilized powder formulation of compound 1

[0438] Compound 1 formulation suitable for injection was prepared according to the procedures outlined below. Vehicle preparation

[0439] A single use formulation vessel is staged.

[0440] Water for Injection is charged at approximately 80% of the final vehicle volume.

[0441] The formula amounts of sodium acetate trihydrate and mannitol are added and dissolved by stirring.

[0442] The pH of the vehicle is adjusted to pH 5.4 ± 0.2 with acetic acid.

[0443] The vehicle is brought to volume by the addition of Water for InjectionAttorney Docket No. T0947.70006WO00 Drug Product preparation

[0444] A single use formulation vessel is staged.

[0445] Vehicle is charged at approximately 80% of the final formulation volume.

[0446] The formula amount of compound 1 is added and dissolved by stirring.

[0447] The pH of the solution is adjusted to pH 5.4 ± 0.2 by the dropwise addition of acetic acid or sodium hydroxide.

[0448] A sample is analyzed for concentration.

[0449] The result of the analysis is used to calculate the amount of additional vehicle needed to dilute to target concentration.

[0450] The formulation is brought to volume (weight).

[0451] A sample is analyzed to confirm label strength.

[0452] The solution is filtered through a membrane filter.

[0453] A sample is removed for bioburden testing.

[0454] The solution is sterilized by filtering through a series of two membrane filters.

[0455] The solution is then filled into 2R glass vials at a volume of about 0.5 to 1.0 mL.

[0456] Filled vials are partially stoppered and dried by lyophilization.

[0457] The dried drug product is backfilled with nitrogen, fully stoppered, unloaded from the lyophilizer, and crimped.

[0458] The drug product is placed into storage at about 2 to 8 °C. Example 3: Preparation of reconstituted formulation for injection

[0459] Providing a vial containing liquid diluent, another vial containing Compound 1 lyophilized powder, a 1 mL Luer lock syringe, a syringe needle, and two vial adapters suitable for attaching to the syringe.

[0460] Attaching a vial adaptor to the vial containing liquid diluent.

[0461] Drawing all the liquid diluent into the syringe.

[0462] Attaching a vial adaptor to the vial containing Compound 1.

[0463] Injecting the diluent into the vial containing Compound 1.

[0464] Mixing the diluent and compound 1 for at least 60 seconds and letting the mixture sit for at least 60 seconds to form a colorless clear solution free from visible particles.

[0465] Drawing all the mixed solution into the syringe.Attorney Docket No. T0947.70006WO00

[0466] Attaching the needle to the syringe.

[0467] Injecting the mixed solution subcutaneously into the fat tissue in the abdomen or middle area of the thigh. Example 4: Representative Lyo pharmaceutical formulations

[0468] Several exemplary reconstituted pharmaceutical compositions suitable for injection to patients in phase 3 studies are presented in the Tables below (see also Tables 1-3) . The compositions were prepared as described herein by mixing a lyophilized pharmaceutical composition comprising Compound 1 with a liquid diluent prior to injection, wherein the lyophilized pharmaceutical composition is in powder form. Table 23. Composition of reconstituted Compound 1 for injection Component 10mg 15mg 20mg 30mg 45mg 60mg. Component10mg 15mg 20mg 30mg 45mg 60mgTable 25. Zn:Compound 1 molar ratio of reconstituted Compound 1 composition for injection Component10mg 15mg* 20mg 30mg 45mg 60mgAttorney Docket No. T0947.70006WO00 Zinc acetate (mM) 4.9 4.9 4.9 4.9 4.9 4.9 Zn:Compound 1 molar ratio 1.22:1 0.80:1 0.61:1 0.41:1 0.27:1 0.20:1ComponentQuantity per vialC m nd 1 212% ExampleStudy Design

[0469] Below is an open label, randomized, crossover study in two groups of 16 healthy subjects each. The study involves a PFS formulation, which comprises compound 1 dissolved in an aqueous NaCl solution. A pre-filled syringe containing the PFS formulation is used for patient injections.

[0470] Table 27 presents the results of the open-label, randomized, crossover study conducted with 16 healthy subjects in Group 1. Table 28 shows the findings of an open-label, randomized, crossover study involving another 16 healthy subjects in Group 2. In both Tables 27 and 28, the patients received the PFS formulation for Treatment 1. Table 27. Group 1. Compound 1 Zn:Compound Dosing Volume ConcentrationAttorney Docket No. T0947.70006WO00 Table 28. Group 2. Compound 1 Zn:Compound Dosing Volume Concentration Treatment Dose (mg) 1 Ratio (mL) (mg / mL)re of lyophilized Compound 1. Figure 1A shows the changes of mean plasma concentration of Compound 1 over time in patients who were administered with different doses of the compound. Figure 1B shows changes of median plasma concentration of Compound 1 over time in patients who were administered with various doses of the compound.

[0472] Figure 2A shows the exposure of the formulation containing lyophilized Compound 1 is dose related. Figure 2B demonstrates the exposures of individual subjects to different doses of Compound 1.

[0473] Figure 3A shows the comparisons of the mean plasma concentrations of Compound 1 for patients in groups 1 and 2 who were administered with PFS formulations and those who received reconstituted lyophilized formulations containing 20 mg of Compound 1.

[0474] Figure 3B shows the comparisons of the median plasma concentrations of Compound 1 for patients in groups 1 and 2 who were administered with PFS formulations and those who received reconstituted lyophilized formulations containing 20 mg of Compound 1.

[0475] Both higher mean and median plasma concentrations of Compound 1 were observed in patients who were administered with reconstituted lyophilized pharmaceutical compositions compared to those who received PFS formulations. For example, patients in groups 1 and 2 who were administered with reconstituted lyophilized formulations had mean maximum plasma Compound 1 concentrations of about 240 to about 260 ng / mL , whereas patients who received PFS formulations had mean maximum plasma Compound 1 concentrations of about 110 to about 130 ng / mL. Similarly, patients in groups 1 and 2 who were administered with reconstituted lyophilized formulations had median maximum plasma Compound 1 concentrations of about 220 to about 250Attorney Docket No. T0947.70006WO00 ng / mL , while patients who received PFS formulations had median maximum plasma Compound 1 concentrations of about 110 to about 140 ng / mL.

[0476] Table 29 shows a comparison of Compound 1 exposures in patients who were administered with PFS formulations containing 20 mg of compound 1 and lyophilized formulations containing 20 mg of Compound 1. Table 29. PK data for PFS vs lyophilized formulations with 20 mg Compound 1 in Groups 1 and 2 Dose Tmax Cmax AUC AUC t CL / F Formulation N last inf 1 / 2 (mg) (hr) (ng / mL) (ng·h / mL) (ng·h / mL) (h) (L / h) 1 3d 1 provides higher bioavailability when compared to the PFS formulation.

[0478] Figure 4A shows the comparisons of the mean plasma iron concentrations in patients of groups 1 and 2 who were administered with PFS formulations versus reconstituted lyophilized formulations containing different doses of Compound 1, such as 10 mg, 20 mg, 30 mg, 45 mg, and 60 mg. Similarly, Figure 4B shows the comparisons of the median plasma iron concentrations in patients of groups 1 and 2 who were administered with PFS formulations versus reconstituted lyophilized formulations containing different doses of Compound 1, such as 10 mg, 20 mg, 30 mg, 45 mg, and 60 mg. Table 30. Summary of Lyophilized Compound 1 Pharmacokinetics Dose Tmax Cmax AUClast AUCinf t1 / 2 CL / F NAttorney Docket No. T0947.70006WO00 1053±1333 46381±18276 47448±18035 34.4±13.9 1.47±0.63 60 13 2 (1, 72) (127) (39.4) (40.1) (40.4) (43.0)% coefficient of variation (CV) presented in parenthesis

[0479] Table 30 demonstrates that the lyophilized formulation exhibits higher AUC (area under the curve) values than the PFS formulation at comparable doses, such as 20 mg. Increases in Cmax (maximum concentration) and AUC are not proportional to the dose. Additionally, at higher doses, Tmax (time to reach maximum concentration) occurs earlier than at lower dose.

[0480] The experimental results demonstrated a dose-related increase in Compound 1 exposure with the lyophilized formulation across the dose range of 10 to 60 mg. However, the increase in exposure was not directly proportional to the dose. At 20 mg (the common dose tested), the lyophilized formulation exhibited about 1.6-fold higher exposure compared to the PFS formulation. The pharmacokinetics of 20 mg Compound 1 were comparable between Groups 1 and 2. Example 6: Lyophilized Formulation Stability Studies

[0481] Stability studies were conducted to determine the shelf-life of Compound 1 for injection at dose strength of 10 mg, 15 mg, 20 mg, 30 mg, 45 mg and 60 mg were performed. Analytical procedures

[0482] A^UHPLC^analytical procedure^has been^validated for release and stability testing of PTG- 300 for Injection. Forced degradation data by light (ICH Q1B) and heat (50°C) exposure verify that method is stability indicating by confirmation of main peak homogeneity by photodiode array assessment. Evaluation

[0483] Evaluation of data at all dose strengths shows minimal signs of degradation at the 2-8°C long term and accelerated 25°C / 60% RH storage conditions. The stressed 40°C / 75% RH storage conditions show multiple signs of degradation, particularly in the UHPLC impurity profile; however, none of the values are out of the specified ranges.Attorney Docket No. T0947.70006WO00

[0484] The primary degradant observed at all storage conditions is Iso-Asp. Minor increases in Dimer 1, Dimer 2 and several late eluting species that have not been identified have only been observed thus far at the stressed 40°C / 75% RH storage condition. Conclusions

[0485] Compound 1 for injection will be stored at the 2-8°C storage condition, protected from light, with an interim shelf-life of 18 months.

[0486] Figure 5A shows the stability data of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 at 5 ºC over a period of approximately 24 months. The results indicate that the lyophilized formations have excellent long-term stability, with purity level greater than 98% after storage at 5 ºC for over 18 months. The lyophilized formulations containing 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 (see Tables 23-25) demonstrate even greater stability, maintaining purity level above 99% after storing at 5 ºC for over 18 months.

[0487] Figure 5B shows a comparison of the stability of a PFS liquid formulation containing 30 mg Compound 1 and a reconstituted lyophilized formulation containing 30 mg of Compound 1 at 5 ºC over a period of approximately 24 months. The results indicate the lyophilized formulation has superior long-term storage stability compared to the PFS formulation, as demonstrated by the purity profiles at various storage times. For example, at 18 months, the PFS formulation has purity level slightly over 98%, whereas the lyophilized formulation maintains a purity level near 99%.

[0488] Figure 5C shows the stability data of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 at 25 ºC over a period of approximately 24 months. The results indicate that the lyophilized formations have long-term stability as demonstrated by having purity level greater than 98% after being stored at 25 ºC for over 18 months.

[0489] Figure 5D shows a comparison of the stabilities of a PFS liquid formulation containing 30 mg of Compound 1 and reconstituted lyophilized formulation containing 30 mg of Compound 1, stored at 25 ºC over a period of approximately 24 months. The results indicate the lyophilized formulation has superior long-term storage stability when compared to the PFS formulation as demonstrated by the purity profiles at various storage times. For example, at 18 months, the PFS formulation has purity level slightly over 98%, whereas the lyophilized formulation maintains a purity level close to 99%.Attorney Docket No. T0947.70006WO00

[0490] Figure 6A illustrates the stability data of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg or 60 mg of Compound 1 at 5 ºC over a period of approximately 24 months. The results indicate that the lyophilized formations have excellent long-term stability. For example, the lyophilized formulations containing 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 have less than 0.8% of Asp related degradant after being stored at 5 ºC for 24 months. Similarly, the lyophilized formulation containing 10 mg of Compound 1 shows less than 1.5% of Asp-related degradant after storage at 5 ºC for 18 months. These findings confirm the reliability and durability of the lyophilized formulations.

[0491] Figure 6B shows a comparison of the stability of a PFS liquid formulation containing 30 mg of Compound 1 and a reconstituted lyophilized formulation containing 30 mg of Compound 1, both stored at 5 ºC over a period of approximately 24 months. The data clearly show that the lyophilized formulation has superior long-term storage stability compared to the PFS formulation, as evidenced by the presence of a major Asp-related degradant at various storage times. For example, after 18 months, the PFS formulation has less than 1.4% of the Asp related degradant, whereas the lyophilized formulation has less than 0.8% of the Asp-related degradant at the same storage conditions. Furthermore, the lyophilized formulation maintains the level of the Asp-related degradant below 0.8% after 24 months of storage at 5 ºC. These findings indicate the robustness and reliability of the lyophilized formulation in maintaining the compound’s stability over an extended period.

[0492] Figure 6C shows the stability data of Compound 1 lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1, at 25º C over a period of approximately 24 months. The results demonstrate that the long-term stability of the lyophilized formations, with Asp-related degradant in the range of 0.8% to 1.8% after 24 months of storage at 25 ºC. For example, the lyophilized formations exhibit Asp-related degradant in the range of 0.8% to 1.7% after being stored at 25 ºC for 18 months. Furthermore, the lyophilized formulations containing 15 mg or 30 mg of Compound 1 maintain less than 0.8% -1% of Asp-related degradant after being stored at 25 ºC for 18 months. These results highlight the effectiveness of the lyophilized formulations in preserving the stability and quality of Compound 1 over an extended period even at elevated temperatures.

[0493] Figure 6D shows a comparison of the stabilities of a PFS liquid formulation containing 30 mg of Compound 1 and a lyophilized formulation containing 30 mg of Compound 1 at 25 ºC over aAttorney Docket No. T0947.70006WO00 period of approximately 24 months. Compared to the PFS formulation, the results clearly indicate that the lyophilized formulation exhibits significant long-term storage stability as evident from the presence of Asp-related degradant at various storage times. For example, after 24 months of storage at 25 ºC, the lyophilized formulation maintains Asp-related degradant at 1% or lower, demonstrating its robust stability under these conditions. In contrast, the PFS formulation experiences a substantial increase in Asp-related degradant, exceeding 5% after storage at 25 ºC for about 6 months. These findings emphasize the superior stability of the lyophilized formulation over extended periods and challenging storage conditions. Example 7: Comparison of Stable Liquid Formulations and Lyophilized Formulations in Clinical Studies

[0494] Tables 31-34 below provide exemplary stable liquid pharmaceutical compositions containing 20 mg and 45 mg of peptide of formula (I) (Compound 1) suitable for injection to patients in clinical studies. The formulations were prepared as described herein by mixing Compound 1 with excipients resulting in an aqueous solution for use in treatment groups B, C, D, E, F and G. Compound 1 and excipients in Tables 31 and 32 are provided in mg / mL. Compound 1 and excipients in Table 33 are shown in percentages. The molar ratios of zinc and compound 1 in the formulations are shown in Table 34. Table 31. Stable Liquid Formulations of compound 1 for injection (mg / mL) Composition (mg per mL)Attorney Docket No. T0947.70006WO00 Table 32. Formulation of compound 1for injection (mg / dose) Composition (mg per dose) Component Treatment Treatment Treatment Treatment Treatment Treatmenta e . ormua on o compoun or njec on (w / w) ( ) Composition (mg per dose) tAttorney Docket No. T0947.70006WO00 Table 34. Formulation of compound 1 for injection with Zinc Chloride (molar ratio) Composition (mg per mL) Component Treatment Treatment Treatment Treatment Treatment Treatmentformulations containing different ratios of zinc chloride to Compound 1, relative to Compound 1 lyophilized formulation and Compound 1 PFS formulation following subcutaneous administration. A Compound 1 PFS formulation comprises compound 1 dissolved in an aqueous NaCl solution. Study Design

[0496] An open-label, randomized study to characterize the pharmacokinetics of Compound 1 stable liquid formulations with varying ratios of zinc chloride to Compound 1 at two dose levels, relative to the corresponding Compound 1 lyophilized product following subcutaneous administration in two groups of healthy subjects. Plasma samples were collected to analyze the pharmacokinetic profile of Compound 1. The selected doses for the studies were 20 mg and 45 mg, administered subcutaneously. Treatment assignment

[0497] This was a single center, open-label, cross-over study in two (2) groups of 16 healthy subjects each. Subjects were screened for eligibility within 28 days prior to the first treatment.

[0498] Subjects in Group 1 received the following 4 treatments:

[0499] Treatment A: Single dose of reconstituted 20 mg Compound 1 lyophilized product.

[0500] Treatment B: Single dose of 20 mg Compound 1 stable liquid formulation with 2:1 molar ratio of zinc chloride to Compound 1.Attorney Docket No. T0947.70006WO00

[0501] Treatment C: Single dose of 20 mg Compound 1 stable liquid formulation with 1:1 molar ratio of zinc chloride to Compound 1.

[0502] Treatment D: Single dose of 20 mg Compound 1 stable liquid formulation with 0.75:1 molar ratio of zinc chloride to Compound 1.

[0503] Subjects were randomized to one of the following treatment protocols. Four subjects were randomized to each protocols as shown in Table 35. Table 35 Protocol Period 1 Period 2 Period 3 Period 4 [

[0505] Treatment E: Single dose of reconstituted 45 mg Compound 1 lyophilized product.

[0506] Treatment F: Single dose of 45 mg compound 1 stable liquid formulation with 2:1 molar ratio of zinc chloride to Compound.

[0507] Treatment G: Single dose of 45 mg compound 1 stable liquid formulation with 1:1 molar ratio of zinc chloride to Compound 1.

[0508] Treatment H: Single dose of 45 mg compound 1 stable liquid formulation with 0.75:1 molar ratio of zinc chloride to Compound 1.

[0509] Subjects were randomized to one of the following treatment protocols. Four subjects were randomized to each protocol as shown in Table 36. Table 36 Protocol Period 1 Period 2 Period 3 Period 4Attorney Docket No. T0947.70006WO00

[0510] The tables below provide comparisons of Compound 1 stable liquid formulation relative to Compound 1 lyophilized formulation at 20 mg and 45 mg doses. Randomized, open-label, four-way crossover study in 2 groups of 16 healthy subjects. Each subject received a stable liquid formulation on 3 occasions and the corresponding dose of the lyophilized product on 1 occasion. Table 37: 20 mg of Compound 1 Lyophilized Product Stable Liquid Stable Liquid Stable Liquid 0.61:1 Zinc and 0.75:1 Zinc and 1:1 Zinc and 2:1 Zinc and C d 1 M l C d 1 C d 1 C d 1 te, lLyophilized Stable Liquid Stable Liquid Stable Liquid Product 0.75:1 Zinc and 1:1 Zinc and 2:1 Zinc and e,gure sp ays e p armaco ne c pro es o eren qu ormu a ons o Compound 1, each with varying ratios of zinc to Compound 1, along with the lyophilized formulation of Compound 1. The subjects were administered a 20 mg dose of Compound 1. The pharmacokinetic results are summarized in Table 39 and Table 40. As shown in Figure 7, compared to the lyophilized formulation, the liquid formulations of compound 1 have improved bioavailability.

[0512] Table 39 presents a comparison of pharmacokinetic parameters between the liquid formulations of Compound 1 and lyophilized formulation of Compound 1, both administered at a 20 mg dose. The pharmaceutical compositions were given to 16 subjects as described in group 1 above.Attorney Docket No. T0947.70006WO00 Table 39: 20 mg of Compound 1 20 mg Parameters A: Lyo D: SLF 0.75:1 C: SLF 1:1 B: SLF 2:1Lyo: lyophilized formulation Table 40: 45 mg of Compound 1 45 mg Parameters E: Lyo F: SLF 2:1 G: SLF 1:1 H: SLF 0.75:1Lyo: lyophilized formulation

[0513] The mean rusfertide plasma concentration-time profile following dosing for Group 1 is presented graphically in Figure 7. The mean rusfertide plasma concentration-time profile following dosing for Group 2 is presented graphically in Figure 8. Figure 9 demonstrates a comparison of the pharmacokinetics of a PFS formulation, and a stable liquid formulation administered to the subjects. The stable liquid formulation comprises zinc and compound 1 with a molar ratio of 2:1.

[0514] Pharmacokinetic parameters for rusfertide following 20 mg (Group 1) and 45 mg (Group 2) are summarized in Table 39 and Table 40, respectively.

[0515] Following subcutaneous administration, rusfertide concentrations were generally noted by 0.5 hours of administration. The median time to peak concentration was 24 hours for the 20 mg dose of lyophilized formulation and 6 hours for the 45 mg dose. Median peak concentrations for the liquid formulations were 24 to 48 hours over both doses, with the 2:1 Zn chloride:rusfertide molar ratio formulation exhibiting a later Tmax compared to the 0.75:1 formulation. Elimination half-life was generally comparable (17-27 hours) across the two doses for the liquid formulations andAttorney Docket No. T0947.70006WO00 approximately 25 hours for the 20 mg dose of the lyophilized formulation and 45 hours for the 45 mg dose of the lyophilized formulation.

[0516] All stable liquid formulations containing zinc chloride had a higher AUC and Cmax compared to the corresponding dose of the reconstituted lyophilized product. The 0.75:1 Zn chloride:rusfertide molar ratio formulation had some improvement over the lyophilized formulation, the 1:1 formulation had an intermediate improvement over the lyophilized formulation and the 2:1 formulation had the most improvement over the lyophilized formulation at both the 20 mg and 45 mg doses. (Table 39 and Table 40).

[0517] Table 41 below summarizes the relative bioavailability of the liquid formulations relative to the lyophilized formulation. The 20 mg and 45 mg doses of 2:1 Zn chloride:rusfertide molar ratio formulation had an approximately 1.8- and 2.3-fold higher AUC, respectively, compared to the corresponding doses of the lyophilized formulation. Consistent with the plasma profile, the 2:1 molar ratio formulation had the highest improvement over the lyophilized formulation (1.8 to 2.3- fold improvement), the 1:1 formulation an intermediate improvement over the lyophilized formulation (1.6- to 1.9-fold improvement), and the 0.75:1 formulation had the least improvement over the lyophilized formulation (1.3- to 1.7-fold improvement). Table 41: Relative Bioavailability of Stable Liquid Formulations Compared to Lyo (Mean+ / -SD) – PK from Population that Completed the Study Treatment Comparison N Ratio of AUC (%)Example 8: SLF Stability Studies 1

[0518] The same buffer strength, pH, and PTG- 300 concentration were chosen for all formulations. Each formulation was studied with and without zinc chloride (ZnCl2). Aspartic acid (L-Asp) wasAttorney Docket No. T0947.70006WO00 included at two different concentrations. Two different sugars, Trehalose and sucrose, were studied with sucrose studied at three different concentrations.

[0519] Materials

[0520] Glacial acetic acid

[0521] Sodium acetate trihydrate

[0522] Zinc chloride

[0523] Sucrose

[0524] l-aspartic acid

[0525] Trehalose

[0526] Compound 1

[0527] Water, USP / EP

[0528] Formulation Preparation

[0529] i) 25 mM sodium acetate, pH 4.5 was prepared by dissolving 1.7012 g sodium acetate trihydrate in USP water, adjusting the pH with 1 M acetic acid, and bringing the volume to 500 ml. The final pH of the buffer was pH 4.51.

[0530] ii) Stock solutions of each excipient were prepared in the 25 mM sodium acetate buffer, pH 4.5:

[0531] a. 1.9 mg / ml ZnCl2 (14 mM)

[0532] b. 685 mg / ml Sucrose (2 M)

[0533] c. 171 mg / ml Trehalose (500 mM)

[0534] iii) 50 mg / ml PTG-300 was prepared in 25 mM sodium acetate buffer, pH 4.5.4.185 g of Compound 1 (CF = 89.6%, 4.185 g x 0.896 = 3.750 g PTG-300) was added to 70 ml of buffer. The pH was adjusted with 1 M acetic acid, and the total volume was brought to 75 ml.

[0535] iv) Each formulation was prepared by combining excipients. If necessary, the pH of each solution was adjusted with 1 M NaOH before final volume adjustment to 5 ml for each formulation.

[0536] v) Each formulation was filtered through a 0.2 µm filter (Acrodisc 25 mm, 0.2 µm HT Tuffryn membrane) into an 8-ml glass scintillation vial.

[0537] i) The samples were placed in the 40°C / 75%RH stability chamber and removed at the designated timepoints.

[0538] ii) At each timepoint, a 1-ml aliquot of each sample was removed. The appearance and pH were recorded before HPLC analysis.Attorney Docket No. T0947.70006WO00

[0539] i) 250 µl of each formulation was added to vial and brought to 10 ml with USP water.

[0540] ii) Each sample was mixed by inverting the flask several times.

[0541] iii) At t=3M: Each sample was filtered through a 0.2 µm filter (Acrodisc 13 mm, 0.2 µm Supor membrane).

[0542] Results

[0543] No significant change in pH was observed over the course of the stability study. The appearance of the solution did not change until 3 months of storage at 40°C when some formulations developed slight discoloration. Most formulations resulted in significant improvement in total impurities: 7-8% increase in total impurities for Thermostability Study 1 formulations versus 33% increase in total impurities for 50 mg / ml Compound 1 in saline after 1 month at 40°C.

[0544] The addition of disaccharides, which are used to reduce aggregation and disulfide exchange for peptides in solution, did not improve the stability Compound 1 compared to the buffer alone and specifically increased the presence of degradant at RRT 1.15 (chromatographic eluate at relative retention time 1.15). The addition of ZnCl2resulted in improvement for degradants at RRT ≥ 1.27. The same was observed for the addition of L-aspartic acid, but L-aspartic acid also increased the presence of degradants at RRT ~0.88 and ~1.06. Increasing the concentration of L-aspartic acid did not improve stability. The combination of ZnCl2and L-aspartic acid did not have an additive effect on stability. Example 9. SLF Stability Studies 2

[0545] The purpose of the study was to investigate the effect of removing excipient from the liquid formulation and how it impacts on stability of the liquid formulations at different dose strengths. The study provides comparisons of liquid formulations comprising compound 1 and the PFS formulation at each dose strength. The solution stability at 25°C was assessed for improvement as compared to the PFS formulation. Samples were also stored at 5°C to confirm shelf-life projections.

[0546] Table 42 provides a summary of various Compound 1 formulations in the presence or absence of ZnCl2. As shown in Table 42, the PFS formulation has form # C1 and contains sodium acetate trihydrate and NaCl as excipients. The stable liquid formulations (SLF) in Table 42 do not contain any NaCl.Attorney Docket No. T0947.70006WO00 Table 42 sodium PEG Cmpd 1 acetate NaCl Sorbitol ZnCl2Molar RatioPFS: a liquid formulation containing compound 1, NaOAc buffer, NaCl and water.

[0547] Table 43 shows comparisons of liquid formulation comprising compound 1 and ZnCl2and the PFS formulation at a pH of 5.6 and 5 ºC. The results indicate that the stable liquid formulations with a molar ratio of Zn to Compound 1 at 2:1 or 1:1, have 2 to 3 folds less of Asp-related degradants. Table 43 Asp-related Molar ratio ofAttorney Docket No. T0947.70006WO00 B: Form R5 (10 mg / ml): 2.72 mg / ml acetate + 58 mg / ml 1:1 0.27 sorbitol + 18 mg / ml PEG3350 + 1.12 mg / ml ZnCl

[0548] Aining zinc salts showed a reduction in Asp-related degradants and high molecular weight degradants when compared to the PFS formulation. For example, the stable liquid formulations, with a molar ratio of zinc to Compound 1 at 2:1 or 1:1, exhibit 2-fold lower Asp-related degradants compared to the PFS formulation.

[0549] Figure 10A shows the stability data of the PFS formulation and liquid formulations containing a 2:1 ratio of Zn and Compound 1 at 5 ºC over approximately 9 months. The stable liquid formulations comprise 10 mg / mL, 30 mg / mL, 60 mg / mL, or 120 mg / mL of Compound 1. Clearly, when compared to the PFS formulation, the liquid formulations containing the zinc salt exhibit higher purity after storage at 5 ºC for about 9 months.

[0550] Figure 10B presents the Asp-related degradants in the PFS formulation and liquid formulations containing a 2:1 molar ratio of Zn and Compound 1 at 5 ºC over approximately 9 months. Clearly, when compared to the PFS formulation, the liquid formulation containing the zinc salt has lower Asp-related degradants or impurities after storage at 5 ºC for about 9 months.

[0551] Figure 10C illustrates the total degradants in the PFS formulation and liquid formulations containing a 2:1 molar ratio of Zn and Compound 1 at 5 ºC over approximately 9 months. Clearly, when compared to the PFS formulation, the liquid formulation containing the zinc salt has lower total degradants or impurities after storage at 5 ºC for about 9 months.

[0552] Figure 11A illustrates the stability data of the PFS formulation and liquid formulations containing different molar ratios of Zn and Compound 1 at 5 ºC over approximately 9 months. The stable liquid formulations comprise 2:1, 1:1 or 0 molar ratio of zinc and compound 1. Clearly, when compared to the PFS formulation, the liquid formulations containing 2:1 and 1:1 molar ratio of zinc and compound 1 exhibit higher purity after storage at 5 ºC for about 9 months.

[0553] Figure 11B presents the Asp-related degradants in the PFS formulation and liquid formulations containing 2:1, 1:1 or 0 molar ratio of Zn and Compound 1 at 5 ºC over approximately 9 months. Clearly, when compared to the PFS formulation, the liquid formulations containing theAttorney Docket No. T0947.70006WO00 zinc salt have significantly lower Asp-related degradants or impurities after storage at 5 ºC for about 9 months.

[0554] Figure 11C illustrates the total degradants in the PFS formulation and liquid formulations containing 2:1, 1:1 or 0 molar ratio of Zn and Compound 1 at 5 ºC over approximately 9 months. Clearly, when compared to the PFS formulation, the liquid formulations containing the zinc salt have significantly lower total degradants or impurities after storage at 5 ºC for about 9 months. Example 10. Lyophilized Formulation Stability Studies

[0555] Stability studies were conducted to determine the shelf-life of Compound 1 for injection at dose strength of 10 mg, 15 mg, 20 mg, 30 mg, 45 mg and 60 mg were performed. Analytical procedures

[0556] A^UHPLC^analytical procedure^has been^validated for release and stability testing of PTG- 300 for Injection. Forced degradation data by light (ICH Q1B) and heat (50°C) exposure verify that method is stability indicating by confirmation of main peak homogeneity by photodiode array assessment. Evaluation

[0557] Evaluation of data at all dose strengths shows minimal signs of degradation at the 2-8°C long term and accelerated 25°C / 60% RH storage conditions. The stressed 40°C / 75% RH storage conditions show multiple signs of degradation, particularly in the UHPLC impurity profile; however, none of the values are out of the specified ranges.

[0558] The primary degradant observed at all storage conditions is Iso-Asp. Minor increases in Dimer 1, Dimer 2 and several late eluting species that have not been identified have only been observed thus far at the stressed 40°C / 75% RH storage condition. Conclusions

[0559] Compound 1 for injection will be stored at the 2-8°C storage condition, protected from light, with an interim shelf-life of 18 months.

[0560] Figure 5A shows the stability data of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 at 5 ºC over a period of approximately 24 months. The results indicate that the lyophilized formations have excellent long-term stability,Attorney Docket No. T0947.70006WO00 with purity level greater than 98% after storage at 5 ºC for over 18 months. The lyophilized formulations containing 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 demonstrate even greater stability, maintaining purity level above 99% after storing at 5 ºC for over 18 months.

[0561] Figure 5B shows a comparison of the stability of a PFS liquid formulation containing 30 mg Compound 1 and a reconstituted lyophilized formulation containing 30 mg of Compound 1 at 5 ºC over a period of approximately 24 months. The results indicate the lyophilized formulation has superior long-term storage stability compared to the PFS formulation, as demonstrated by the purity profiles at various storage times. For example, at 18 months, the PFS formulation has purity level slightly over 98%, whereas the lyophilized formulation maintains a purity level near 99%.

[0562] Figure 5C shows the stability data of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 at 25 ºC over a period of approximately 24 months. The results indicate that the lyophilized formations have long-term stability as demonstrated by having purity level greater than 98% after being stored at 25 ºC for over 18 months.

[0563] Figure 5D shows a comparison of the stabilities of a PFS liquid formulation containing 30 mg of Compound 1 and reconstituted lyophilized formulation containing 30 mg of Compound 1, stored at 25 ºC over a period of approximately 24 months. The results indicate the lyophilized formulation has superior long-term storage stability when compared to the PFS formulation as demonstrated by the purity profiles at various storage times. For example, at 18 months, the PFS formulation has purity level slightly over 98%, whereas the lyophilized formulation maintains a purity level close to 99%.

[0564] Figure 6A illustrates the stability data of reconstituted lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg or 60 mg of Compound 1 at 5 ºC over a period of approximately 24 months. The results indicate that the lyophilized formations have excellent long-term stability. For example, the lyophilized formulations containing 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1 have less than 0.8% of Asp related degradant after being stored at 5 ºC for 24 months. Similarly, the lyophilized formulation containing 10 mg of Compound 1 shows less than 1.5% of Asp-related degradant after storage at 5 ºC for 18 months. These findings confirm the reliability and durability of the lyophilized formulations.

[0565] Figure 6B shows a comparison of the stability of a PFS liquid formulation containing 30 mg of Compound 1 and a reconstituted lyophilized formulation containing 30 mg of Compound 1, both stored at 5 ºC over a period of approximately 24 months. The data clearly show that the lyophilizedAttorney Docket No. T0947.70006WO00 formulation has superior long-term storage stability compared to the PFS formulation, as evidenced by the presence of a major Asp-related degradant at various storage times. For example, after 18 months, the PFS formulation has less than 1.4% of the Asp related degradant, whereas the lyophilized formulation has less than 0.8% of the Asp-related degradant at the same storage conditions. Furthermore, the lyophilized formulation maintains the level of the Asp-related degradant below 0.8% after 24 months of storage at 5 ºC. These findings indicate the robustness and reliability of the lyophilized formulation in maintaining the compound’s stability over an extended period.

[0566] Figure 6C shows the stability data of Compound 1 lyophilized formulations containing 10 mg, 15 mg, 20 mg, 30 mg, 45 mg, or 60 mg of Compound 1, at 25º C over a period of approximately 24 months. The results demonstrate that the long-term stability of the lyophilized formations, with Asp-related degradant in the range of 0.8% to 1.8% after 24 months of storage at 25 ºC. For example, the lyophilized formations exhibit Asp-related degradant in the range of 0.8% to 1.7% after being stored at 25 ºC for 18 months. Furthermore, the lyophilized formulations containing 15 mg or 30 mg of Compound 1 maintain less than 0.8% -1% of Asp-related degradant after being stored at 25 ºC for 18 months. These results highlight the effectiveness of the lyophilized formulations in preserving the stability and quality of Compound 1 over an extended period even at elevated temperatures.

[0567] Figure 6D shows a comparison of the stabilities of a PFS liquid formulation containing 30 mg of Compound 1 and a lyophilized formulation containing 30 mg of Compound 1 at 25 ºC over a period of approximately 24 months. Compared to the PFS formulation, the results clearly indicate that the lyophilized formulation exhibits significant long-term storage stability as evident from the presence of Asp-related degradant at various storage times. For example, after 24 months of storage at 25 ºC, the lyophilized formulation maintains Asp-related degradant at 1% or lower, demonstrating its robust stability under these conditions. In contrast, the PFS formulation experiences a substantial increase in Asp-related degradant, exceeding 5% after storage at 25 ºC for about 6 months. These findings emphasize the superior stability of the lyophilized formulation over extended periods and challenging storage conditions. Example 11: Preparation of Stable Liquid Formulations of Peptide of Formula (I)

[0568] A formulation suitable for injection to patients, containing the peptide of formula (I) (referred to as Compound 1) was prepared according to the procedures below.Attorney Docket No. T0947.70006WO00

[0569] Excipients and water were added to ~15 mL vehicle. The mixture was whirled to dissolve.

[0570] Compound 1 was weighed out and added slowly to the mixture until all dissolved.

[0571] The pH value was recorded and the pH was adjusted to the target pH (pH ± 0.2).

[0572] QS with vehicle to the final volume in 20-mL volumetric flask.

[0573] The final pH was recorded.

[0574] The solution was filtered, and 1.0 mL aliquots were prepared in 2 mL serum vials, stopper, and cap.

[0575] All aliquots were stored at 2-8°C until all formulations are completed. The date (t0) that samples were placed in stability chambers was recorded.

[0576] Example 9: Stable Liquid Formulations in Clinical Studies Example 12: Drug Concentration Measurements

[0577] The concentration of rusfertide in plasma was analyzed as follows. Aliquots of human plasma (0.100 mL) were pipetted into assigned positions of a 1 mL 96-well plate and 0.0250 mL of the working internal standard (400 ng / mL rusfertide IS in 1:1 acetonitrile:water) was added to the appropriate wells, with blank 1:1 acetonitrile:water added to the matrix blanks. A 0.300 mL aliquot of protein precipitation reagent (2% formic acid in acetonitrile) was added to each well and the samples were vortexed to mix. The samples were centrifuged at 3500 rpm for 5 minutes, and 0.200 mL of the supernatant was transferred to a clean well plate. The samples were dried under a gentle stream of nitrogen at 40°C. The dried extracts were reconstituted with 0.200 mL of 1:1 acetonitrile:water. A 10 μL aliquot was injected onto the liquid chromatography with tandem mass spectrometry (LC-MS / MS) system.

[0578] Prior to sample analysis, the analytical method for the determination of rusfertide concentration in human plasma was validated for selectivity, linearity, reproducibility, recovery, precision, and accuracy over the concentration range of 2.00 ng / mL to 1000 ng / mL. Plasma sample stability was established for up to 379 days at -70°C and 3 freeze-thaw cycles from -70°C / ice bath. Example 13: Preparation of Lyophilized Rusfertide Formulations Including Zinc Salts Table 44: Lyophilized Formulations Formulation Rusfertide Sodium Acetate Excipients and ConcentrationAttorney Docket No. T0947.70006WO00 2 160 20 3.6% (w / v) mannitol, 0.18% (w / v) zinc acetate, 0.09% (w / v) sucrose

[0580] 40 mM Sodium Acetate Buffer Stock Solution

[0581] A 40 mM sodium acetate buffer stock solution was prepared at 100 mL scale as follows:

[0582] 80 mL of WFI was added to a glass beaker containing a magnetic stirrer bar.

[0583] 330 mg of sodium acetate was weighed into a weighing boat and transferred to the compounding beaker whilst stirring until completely dissolved.

[0584] The pH of the solution was measured and adjusted to pH 5.4 ± 0.2 with glacial acetic acid.

[0585] The solution was transferred to a 100 mL volumetric flask, made to volume with WFI and inverted to mix.

[0586] 1% (w / v) Glycine Stock Solution

[0587] A 1 % (w / v) glycine stock solution was prepared at 10 mL scale as follows:

[0588] 8 mL of WFI was added to a scintillation vial containing a magnetic stirrer bar.

[0589] 100 mg of glycine was weighed into a weighing boat, transferred to the compounding vial and stirred until dissolved.

[0590] The solution was transferred to a 10 mL volumetric flask, made to volume with WFI and inverted to mix.

[0591] 1% (w / v) Sucrose Stock Solution

[0592] A 1 % (w / v) sucrose stock solution was prepared at 10 mL scale as follows:

[0593] 8 mL of WFI was added to a scintillation vial containing a magnetic stirrer bar.

[0594] 100 mg of sucrose was weighed into a weighing boat, transferred to the compounding vial and stirred until dissolved.

[0595] The solution was transferred to a 10 mL volumetric flask, made to volume with WFI and inverted to mix.

[0596] 2.2% (w / v) Zinc Acetate Stock Solution

[0597] A 2.2 % (w / v) zinc acetate stock solution was prepared at 10 mL scale as follows:

[0598] 8 mL of WFI was added to a scintillation vial containing a magnetic stirrer bar.

[0599] 263 mg of zinc acetate dihydrate was weighed into a weighing boat, transferred to the compounding vial and stirred until dissolved.Attorney Docket No. T0947.70006WO00

[0600] The solution was transferred to a 10 mL volumetric flask, made to volume with WFI and inverted to mix.

[0601] PTG-300 Solutions

[0602] Two 160 mg / mL PTG-300 formulations were prepared as per the following methods:

[0603] Preparation of Formulation 1

[0604] 5 mL of the 40 mM sodium acetate buffer stock solution was transferred by pipette into a scintillation vial containing a magnetic stirrer bar.

[0605] 200 mg of mannitol was weighed into weighing boats, transferred to the compounding vial and stirred until dissolved.

[0606] 800 μL of 1 % (w / v) glycine stock solution was transferred by pipette to the compounding vial and stirred to mix.

[0607] 1 mL of 2.2 % (w / v) zinc acetate stock solution was transferred by pipette to the compounding vial and stirrer to mix.

[0608] 1.739 g of PTG-300 was weighed into a weighing boat, transferred to the compounding vial and stirred until completely dissolved.

[0609] Preparation of Formulation 2

[0610] 5 mL of the 40 mM sodium acetate buffer stock solution was transferred by pipette into a scintillation vial containing a magnetic stirrer bar.

[0611] 360 mg of mannitol was weighed into weighing boats, transferred to the compounding vial and stirred until dissolved.

[0612] 900 μL of 1 % (w / v) sucrose stock solution was transferred by pipette to the compounding vial and stirred to mix.

[0613] 1 mL of 2.2 % (w / v) zinc acetate stock solution was transferred by pipette to the compounding vial and stirrer to mix.

[0614] 1.739 g of PTG-300 was weighed into a weighing boat, transferred to the compounding vial and stirred until completely dissolved.

[0615] For the two solutions the remaining steps are as follows:

[0616] The pH of the solutions was measured and adjusted to pH 5.4 ± 0.2 using glacial acetic acid.

[0617] The solutions were transferred to 10 mL volumetric flasks and made to final volume with WFI, inverted to mix and returned to the original compounding beakers. The solutions were stirred for 10 minutes to mix.Attorney Docket No. T0947.70006WO00

[0618] The solutions were filtered through a single 0.22 μm PVDF membrane syringe filter.

[0619] The pH of the filtrate solutions was measured.

[0620] The osmolality of the filtrate solutions was measured

[0621] The formulations were then freeze dried to generate the lyophilized product. Example 14. Synthesis of Peptides

[0622] Unless otherwise specified, reagents and solvents employed in the following were available commercially in standard laboratory reagent or analytical grade and were used without further purification. General procedure for solid-phase synthesis of peptides

[0623] Peptides are synthesized utilizing the CEM liberty Blue Microwave assisted peptide synthesizer. Using the Liberty Blue, Fmoc-deprotection is carried out by addition of 20% piperidine in N,N-dimethylformamide (DMF) and then heating to 90° C using microwave irradiation for 80 sec. After DMF washes the Fmoc-protected amino acids are coupled by addition of 0.2 M amino acid (5 eq), 0.5 M N,N’-Diisopropylcarbodiimide (DIC; 5 eq) and 1.0 M ethyl cyanohydroxyiminoacetate (Oxyma; 5 eq) all dissolved in DMF. The coupling solution is heated using microwave radiation to 90° C for 160 sec. A second coupling is employed when coupling Arg or other sterically hindered amino acids. When coupling with histidine, the reaction is heated to 50° C for 10 min, double coupling. The cycles are repeated until the desired peptide was obtained. General procedure for cleavage of peptides off resin

[0624] Side chain deprotection and cleavage off the resin for peptide analogues disclosed herein is achieved by stirring dry resin in a solution containing trifluoroacetic acid (TFA), water, 2,2’- (ethylenedioxy)diethanethiol (DODT) and triisopropylsilane (TIPS) (90:5:2.5:2.5) for 2 h. Following TFA removal, peptide is precipitated using ice-cold diethyl ether. The solution is centrifuged (5,000 rpm, 4° C, 5 min), and the ether is decanted. The peptide is dissolved in an acetonitrile (MeCN) / water solution (1:1) and the resulting solution is filtered. The linear peptide quality was assessed using electrospray ionization mass spectrometry (ESI-MS).

[0625] Purification of the peptides disclosed herein is carried out using reverse-phase high performance liquid chromatography (RP-HPLC). A semi-preparative column - Gemini 5 μm C18 column (21.2 mm × 250 mm) (Phenomenex®) is applied at the flow rate of 20 mL / min. TheAttorney Docket No. T0947.70006WO00 separation is achieved using linear gradients with mobile phase A (MPA = 0.1% TFA in water) and mobile phase B (MPB = 0.1% TFA in acetonitrile).

[0626] It should be understood that the above description and Examples are only representative of illustrative embodiments and examples. For the convenience of the reader, the above description has focused on a limited number of representative examples of all possible embodiments, examples that teach the principles of the disclosure. The description has not attempted to exhaustively enumerate all possible variations or even combinations of those variations described. That alternate embodiments may not have been presented for a specific portion of the disclosure, or that further undescribed alternate embodiments may be available for a portion, is not to be considered a disclaimer of those alternate embodiments. One of ordinary skill will appreciate that many of those undescribed embodiments, involve differences in technology and materials rather than differences in the application of the principles of the disclosure. Accordingly, the disclosure is not intended to be limited to less than the scope set forth in the following claims and equivalents. Example 15. Introduction

[0627] Polycythemia vera (PV) is a chronic myeloproliferative neoplasm driven by activating mutations in the Janus kinase 2 (JAK2) gene that results in unrestrained erythrocytosis and increased hematocrit and hemoglobin concentrations, placing patients at increased risk of thrombotic events and mortality. Treatment guidelines for PV recommend maintaining hematocrit below 45% to minimize thromboembolic and cardiovascular risk. Treatment is generally initiated with low-dose aspirin and periodic therapeutic phlebotomy to maintain a hematocrit <45%. In patients younger than 60 years and those without a history of a thrombotic event, who have low-risk disease, Pegylated interferon alpha-2a or ropeginterferon alfa-2b may be considered in patients younger than 60 years and those without a history of a thrombotic event, who have low-risk disease, and who have frequent phlebotomies, severe pruritus, persistent symptoms or symptomatic splenomegaly. High-risk disease patients defined as those older than 60 years or with a history of thrombosis are treated with hydroxyurea, a JAK-2 inhibitor, or pegylated interferon alpha. However, current treatments are often ineffective for maintaining hematocrit levels in the recommended range. Among high-risk PV patients, only 25% had hematocrit levels consistency below 45%, while 7% had hematocrit values always >50%.Attorney Docket No. T0947.70006WO00

[0628] Iron is an essential trace element for nearly every living organism and the most abundant trace element in humans. The human body contains approximately 3 to 4 g of iron, with most in erythrocyte hemoglobin (~2-3 g). Hepcidin, a 25-amino acid peptide hormone synthesized primarily by hepatocytes, is the master regulator of systemic iron homeostasis. Hepcidin inhibits iron absorption in the proximal small intestine and controls iron export to the plasma by inducing degradation of the iron exporter ferroportin in macrophages and hepatocytes. Iron and hepcidin regulate each other in a classical endocrine feedback loop. When hepcidin concentrations are low, iron enters blood compartment; when hepcidin concentrations are high, ferroportin is internalized and iron is trapped in enterocytes, macrophages, and hepatocytes, and can lead to iron-restricted anemia. This observation suggests that hepcidin could be a therapeutic useful for treating conditions of excessive erythropoiesis, such as polycythemia vera. However, synthesis of full-length hepcidin is relatively inefficient, and the short plasma half-life of hepcidin due to proteolysis and renal clearance limits its use as a therapeutic agent.

[0629] Rusfertide (also known as PTG-300) is a synthetic peptide mimetic of the natural peptide hormone hepcidin that is being investigated as a potential treatment for PV. The pharmacokinetic and pharmacodynamic characteristics of rusfertide make it suitable as a therapeutic. In a cell-based ferroportin internalization assay, the EC50values of hepcidin and rusfertide were 67.8 nM and 6.12 nM, respectively. Subcutaneous (SC) administration of rusfertide in cynomolgus monkeys results in a dose-dependent reduction of serum iron and anemia. A single SC injection of rusfertide 0.3 mg / kg in cynomolgus monkeys resulted in a maximum reduction in serum iron of 54.9% 24 hours post- dose. Maximum rusfertide plasma concentrations (172±31 nM) were noted at 8 hours post-dose, and the estimated elimination half-life was approximately 24 hours.

[0630] Rusfertide has shown efficacy in PV in Phase 2 studies, reducing elevated hematocrit levels and in maintaining hematocrit levels <45% in patients with PV and recent reports have demonstrated that rusfertide treatment can provide long-term control of hematocrit. Rusfertide is currently being investigated in a randomized, double-blind, placebo-controlled phase 3 study in patients with PV. In addition, a proof-of-concept clinical study has indicated that rusfertide was also well tolerated and effective for controlling iron overload in patients with HFE-related hemochromatosis.

[0631] The dose-ranging pharmacokinetics and pharmacodynamics of an aqueous prefilled syringe formulation of rusfertide that was used in phase 2 studies in patients with PV has been reported previously. Following subcutaneous administration of the aqueous formulation in healthy subjects,Attorney Docket No. T0947.70006WO00 rusfertide plasma concentrations are noted within one hour of dosing, and peak concentrations (Cmax) occurred at a median of approximately 2 to 24 hours. Following the peak, rusfertide concentrations for the aqueous formulation decreased with an elimination half-life of 17.9 to 52.5 hours. Peak plasma concentrations and area under the curve increased with rusfertide dose but less than dose- proportionally. Corresponding with the pharmacokinetics, dose-related pharmacodynamic decreases in serum iron and transferrin-iron saturation were noted.

[0632] While the aqueous formulation was suitable for phase 2 safety and efficacy studies, it had limited stability and was not considered suitable for large global studies. Lyophilization is frequently used to prepare peptides in a solid state, increasing chemical and physical stability. The present study evaluated the dose-ranging pharmacokinetics, pharmacodynamics and tolerability of a lyophilized formulation of rusfertide compared to the aqueous formulation used in previous phase 2 trials. Methods

[0633] The trial protocol, consent form, and other information provided to participants were reviewed and approved by the Advarra Institutional Review Board. The trial was conducted in compliance with the ethical principles originating in or derived from the Declaration of Helsinki and in compliance with all International Council for Harmonization Good Clinical Practice Guidelines, and all study procedures were conducted by scientifically and medically qualified personnel. Written informed consent was obtained from each participant before any study-specific activity was performed. Study Design

[0634] In this randomized, open-label, dose-ranging, four-way crossover study, the pharmacokinetics, pharmacodynamics, safety, and tolerability of single doses of rusfertide were evaluated in two groups of 16 healthy subjects at a single U.S. site using two formulations: a lyophilized powder formulation for reconstitution or an aqueous formulation in a prefilled syringe. The study was conducted between January and March 2022. Subjects in Group 1 received SC doses of rusfertide 20, 30, and 60 mg as a lyophilized formulation and 20 mg as the aqueous formulation in a randomized fashion. Subjects in Group 2 received SC doses of rusfertide 10, 20, and 45 mg as a lyophilized formulation and 20 mg as an aqueous formulation in a randomized fashion. Subjects in both groups received 20 mg of both formulations. This study design was chosen to provide a direct comparison of the 20 mg dose of both formulation and to limit the number of treatments in eachAttorney Docket No. T0947.70006WO00 subject to four treatments to reduce the number of dropouts. The lyophilized formulation was reconstituted by clinical staff using the accompanying diluent and administered to subjects. All dose injections were 0.5 mL in volume, administered subcutaneously to the abdomen, 2 inches from the navel, with a washout period of at least 13 days between treatments. This study did not meet the definition of “applicable clinical trial”, and it was not required to be registered on clinicaltrials.gov. Study Population

[0635] Eligible subjects were male or female, aged 18 to 65 years, inclusive, in good general health with no significant abnormalities on physical examination, with no laboratory values considered clinically significant by the investigator, and a body mass index between 18 and 32 kg / m2. Women were surgically sterile, postmenopausal, or, if of childbearing potential, agreed to use medically acceptable contraception (<1% annual failure rate) during the study and for 30 days after the last dose of study medication. Key exclusion criteria included history of clinically significant endocrine, neurological, gastrointestinal, cardiovascular, hematological, hepatic, immunological, renal, respiratory, or genitourinary abnormalities or disorders, noninvasive squamous cell carcinoma of the skin (unless adequately treated), history of invasive malignancies within the previous 5 years (except localized cured prostrate or cervical cancer), history of severe allergic or anaphylactic reactions, supine blood pressure outside 90 to 139 mm Hg systolic and 50 to 89 mm Hg diastolic, or heart rate greater than100 beats per minute. Dose Selection

[0636] Single doses of 10, 20, 30, 45, and 60 mg of the lyophilized rusfertide formulation were chosen for this study to study the complete range of rusfertide dose strengths that are included in the larger phase 3 safety and efficacy study. The 20 mg aqueous formulation dose strength was included for comparison with the 20 mg lyophilized formulation since 20 mg is the recommended starting dose of rusfertide. Pharmacokinetic Assessments

[0637] To assess the pharmacokinetic (PK) profile of rusfertide and its metabolites, plasma samples were collected predose, and at 1, 2, 4, 8, 12, 24, 36, 48, 72, 96, 120, 144, 168, 192, and 216 hours following dosing. Subjects were released from the clinical unit following the 48-hour sample, with subsequent samples collected at outpatient visits. Plasma samples were isolated by centrifugation and stored at -70°C within 60 minutes of collection.Attorney Docket No. T0947.70006WO00

[0638] Twelve subjects had detectable predose rusfertide plasma concentrations during at least one of the treatments. For these subjects, prior to conducting PK analyses, the plasma concentrations were corrected assuming a first-order elimination using the average estimated elimination half-life estimated following intravenous administration of rusfertide from a separate trial.

[0639] Pharmacokinetic parameters were estimated by noncompartmental methods using Phoenix WinNonlin version 8.3 (Certara, Princeton NJ). Peak plasma concentration (Cmax) and time to peak plasma concentration (Tmax) were observed values. The elimination rate was estimated from the slope of the least-squares regression on the terminal log-linear phase. Area under the plasma concentration-time curve from time zero to the last quantifiable concentration (AUCt) was estimated by a linear trapezoidal method and was extrapolated to infinity (AUCinf) by dividing the last quantifiable concentration by the elimination rate. To allow comparison across treatments in this crossover study conducted in two groups of subjects, summaries of pharmacokinetic data are based on the group of subjects that completed all treatments.

[0640] The pharmacokinetic analysis set included all subjects who received at least one dose of study medication and had data for at least one PK parameter. PK completers were subjects who had received all four study doses. Pharmacokinetic parameters were summarized using descriptive statistics. Bioanalytical Methods

[0641] Plasma concentrations of rusfertide and metabolites (M1, M4, M6, M9) were determined using two liquid chromatography / tandem mass spectrometry (LC-MS / MS) methods that were performed under Good Laboratory Practice guidelines: a multianalyte LC-MS / MS method for rusfertide, M1, M6, and M9; and a separate LC-MS / MS method for M4. The bioanalytical methods were developed in accordance with current ICH guidelines on bioanalytical method validation and study sample analysis.

[0642] The analytical methods for determination of rusfertide and M4 concentrations were validated for selectivity, linearity, reproducibility, recovery, precision, and accuracy over the concentration range of 2.00 to 500 ng / mL and qualified for M1, M6, and M9 over a concentration range of 3.00 to 750 ng / mL.

[0643] Quality control cumulative accuracy (percent relative error [RE]) was -2.0% to 3.0% for rusfertide and -12.6% to 6.7% for the metabolites. Cumulative precision (percent coefficient of variation [CV]) was ≤3.8% for rusfertide and ≤10.4% for the metabolites.Attorney Docket No. T0947.70006WO00 Pharmacodynamic Assessments

[0644] The pharmacodynamic (PD) effects of rusfertide were assessed by measuring serum iron and transferrin-iron saturation (TSAT). Samples were collected predose, and at 4, 8, 24, 48, 72, 96, 120, 144, and 168 hours following dosing.

[0645] Pharmacodynamic effects on serum iron and TSAT were summarized by estimating the area under the pharmacodynamic effect-time curve (AUEC) from zero to 168 hours. The rusfertide AUC- AUEC relationship for serum iron and for TSAT was investigated. All subjects who received at least one dose of study medication and had AUEC data for at least one PD endpoint were included in the PD analyses. Safety Assessments

[0646] Safety evaluations were based on adverse events (AEs) and the use of concomitant medications, as well as clinical laboratory test values, physical examinations, vital sign measurements, and electrocardiogram (ECG) findings, each assessed at prespecified time points.

[0647] Adverse event occurrences and concomitant medication usage were recorded throughout the study. Physical examinations were conducted at screening and at the end of the study. Clinical laboratory assessments, including hematology, clinical chemistry, coagulation, and urinalysis, were conducted at check-in and at 4 and 48 hours following dosing, and at the end of the study. Electrocardiogram measurements and vital sign recordings were made predose, and at 2, 4, 8, 12, 24, 48, 72, 120, and 216 hours following dosing.

[0648] All participants who received at least one dose of study medication were included in the safety analyses. Adverse events were considered treatment emergent if they started or worsened after the first administration of treatment. Severity grading of AEs was done according to the Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0. AEs were coded using the Medical Dictionary for Regulatory Activities (MedDRA; Version 24.1) and evaluated by the principal investigator. Intensity of the AE was rated as mild, moderate, severe, life threatening or fatal, and the relationship between the AE and study medication was indicated as not related, unlikely related, possibly related, or related. Immunogenicity Assessments

[0649] Serum samples were collected prior to the first rusfertide dose and at the end of study participation for determination of rusfertide antidrug antibodies. Serum antidrug antibodies wereAttorney Docket No. T0947.70006WO00 examined via a validated enzyme-linked immunosorbent assay using a three-tiered approach (screening, confirmation, and titration analysis). Statistical Analyses

[0650] Since this study was not a bioequivalence study, no formal sample size estimations were performed in this single dose trial. The number of participants enrolled in each group was consistent with the customary size employed in pharmacokinetic studies and was expected to allow meaningful assessment of pharmacokinetics and pharmacodynamics and clinical judgement of safety and tolerability.

[0651] Statistical analyses were performed using SAS version 9.4 (SAS Institute, Cary, NC). Dose proportionality of Cmax, AUCt, and AUCinf for the rusfertide lyophilized formulation was assessed based on a power model [Y=α·(dose)β] as described by Gough et al using the modification by Smith et al. In the power model, Y is the pharmacokinetic parameter, α is the expected value of Y for a reference dose, and β is the proportionality exponent. A mixed-effects model, allowing for random between-subject variability in α and β, was implemented to estimate the proportionality constant and its 90% confidence interval (CI). Dose proportionality was declared if the calculated 90% CI lay within the acceptance range [1 + ln(ΘL) / log(R), 1 + ln(ΘH) / log(R)], where ΘL and ΘH are the lower and upper limits of the CI (0.8 and 1.25, respectively) and R is the ratio between the highest and lowest doses (R=6 in this study). Results Subject Disposition

[0652] Two groups of 16 subjects were enrolled. Eleven subjects completed all four treatments in Group 1; one subject was withdrawn for protocol noncompliance, one withdrew due to a family emergency, and three discontinued due to AEs. One subject experienced injection site erythema, induration, and itching, another experienced injection site erythema, and a third reported urticaria. Fourteen subjects completed all four treatments in Group 2; one subject discontinued due to COVID 19 infection and another withdrew to care for a COVID-19-infected relative. Overall, 25 of 32 subjects enrolled (78.1%) completed the study. Demographics and Baseline Characteristics

[0653] The demographics and baseline characteristics of subjects in the two groups are summarized in Table 45. Subjects ranged in age from 28 to 63 years and were equally distributed by gender. The majority of subjects were Black or African American (56%), and 31% were White. The mean bodyAttorney Docket No. T0947.70006WO00 mass index (BMI) was 27.2 kg / m2. Subject demographics were generally well balanced between the two groups. Table 45: Subject Demographics and Baseline Characteristics Group 1 Group 2 Total (N=16) (N=16) (N=32)BMI, body mass indexPharmacokinetics

[0654] The mean rusfertide plasma concentration-time profiles following single doses of rusfertide for both formulations are presented in Figure 12. A summary of rusfertide pharmacokinetics is presented in Table 46. Following SC administration, rusfertide concentrations were detected within 1 hour of dosing, with plasma concentrations increasing and sustained over time. The median time to peak rusfertide concentration (Tmax) for the lyophilized formulation was 24 hours for doses up to 30 mg, and 2 to 4 hours for the 45 and 60 mg doses. The median time to peak rusfertide concentration for the 20 mg dose of the aqueous formulation was 1 hour. For doses of 20 mg and higher of the lyophilized formulation, there was a rapid increase in rusfertide concentration; in some instances, a dual peak could be seen in the rusfertide concentration profile. Dose-related increases were seen in rusfertide Cmax and AUC. The mean elimination half-life for rusfertide ranged from 20 to 57 hours. Mean apparent plasma clearance of rusfertide for the lyophilized formulation ranged fromAttorney Docket No. T0947.70006WO00 approximately 0.63 to 1.5 L / h, with apparent clearance increasing with increasing rusfertide dose. The apparent clearance for 20 mg rusfertide as the aqueous formulation was 1.5 L / h compared to an apparent clearance of 0.98 L / hr for the lyophilized formulation. The 20 mg rusfertide lyophilized formulation had an approximate 1.5-fold higher AUCinf exposure than the 20 mg dose of the aqueous formulation. Table 46: Rusfertide Pharmacokinetic Parameters Following Subcutaneous Dosing Aqueous Lyophilized Formulation FormulationAttorney Docket No. T0947.70006WO00

[0655] Rusfertide dose proportionality was evaluated for the lyophilized formulation doses using a power analysis (Table 47, Figure 14). Following single-dose administration of rusfertide, rusfertide Cmax and AUCinf values increased in a less than dose-proportional manner. The proportionality exponent (β) for Cmax and AUCinf were less than unity. The acceptance interval for dose proportionality for this study was (0.8755, 1.12450). The 90% CI of β was not wholly contained within this interval, indicating a less than dose-proportional increase in Cmax and AUC. Table 47: Rusfertide Proportionality Analysis Pharmacokinetic Proportionality Exponent (β) Parameter Value (SE) 90% Confidence Interval

[0656] Metabolite M9 exposure was 24.9% of all drug-related exposure (AUC) for the aqueous formulation and ranged from 25.4% to 32.6% of all drug-related exposure for the lyophilized formulation (Table 46). Metabolite M4 AUC exposure was 20.9% of all drug-related exposure in the aqueous formulation, while in the lyophilized product, the percentage of M4 exposure relative to all drug-related exposure increased with dose, ranging from 1.9% at 10 mg to 18.5% at 60 mg. Therefore, M9 is considered a major metabolite (≥10%) and conservatively, M4 is also considered a major metabolite for this single-dose study. Across all the dose levels, Tmax of M4 and M9 appeared later than that of rusfertide. The elimination half-lives for M4 and M9 were comparable to that for rusfertide. Metabolites M1 and M6 were detected only sporadically and are considered minor metabolites, comprising <1% of all drug-related AUC. Pharmacodynamics

[0657] A dose-related acute decrease in serum iron levels was seen following SC rusfertide (Figure 13A). Peak decrease in serum iron was noted at 24 hours and at higher doses, the decrease was sustained until 72 hours after which serum iron levels started to return to baseline. A dose-related recovery of the pharmacodynamic effect to baseline was noted, with a faster recovery following 10 mg and a slower recovery following 60 mg. At 10 mg, there was an overshoot in the effect above baseline, reflective of the transient effect at this dose and the general variability in the response. The 20 mg dose of the lyophilized formulation had a similar reduction in serum iron compared with the 20 mg aqueous formulation with a similar return to baseline (Figure 13A).Attorney Docket No. T0947.70006WO00

[0658] Consistent with the effects seen with serum iron, TSAT decreased following SC administration, with peak effects noted at 24 to 48 hours (Figure 13B), and the effect was sustained at the higher doses. Following the nadir, TSAT levels returned towards baseline and for the lower doses of 10 and 20 mg the TSAT levels were essentially back to baseline by 168 hours. The 20 mg doses of the lyophilized formulation and the aqueous formulation had a similar pharmacodynamic effect on TSAT.

[0659] The pharmacodynamic effects on serum iron and TSAT generally matched the rusfertide plasma concentration profile. The relationship between AUEC for serum iron and for TSAT and rusfertide AUC was well described by a linear relationship (Figure 15). A comparison of the y- intercept and slope for the linear relationships for the lyophilized formulation and the aqueous formulation for serum iron and for TSAT indicated that the slopes and intercepts between the two formulations were not significantly different (p=0.427 and 0.503 for slope and intercept, respectively, for serum iron, and p=0.195 and p=0.379 for slope and intercept, respectively, for TSAT) and a single linear relationship could be used for both formulations (Table 48). Table 48: Summary of Relationship between Rusfertide Area Under the Curve and Pharmacodynamic Area Under the Effect Curve for Serum Iron and Transferrin-Iron Saturation Intercept Slope

[0660] Sixteen (50%) of the 32 subjects enrolled in the study experienced a treatment-emergent adverse event (TEAE) [Table 49]. No serious TEAEs were reported. Most TEAEs were mild or moderate in severity. One subject in each of the 20 mg aqueous formulation and 60 mg lyophilized formulation experienced injection site erythema that was considered severe. Three subjects experienced treatment-related AEs that led to discontinuation of treatment; injection site erythema, induration, and pruritus in one subject, injection site erythema in a second subject, and urticaria in the third. TEAEs reported in at least two subjects that were considered treatment-related wereAttorney Docket No. T0947.70006WO00 injection site erythema (13%), injection site pruritus (13%), injection site induration (9%), injection site pain (9%). Overall, 9 of 32 subjects (28%) reported injection site reactions. Table 49: Summary of Treatment-Emergent Adverse Events in Two or More Subjects Overall Following Subcutaneous Dosing of Rusfertide as a Lyophilized Formulation and as a Prefilled Syringe Aqueous Formulation Lyophilized Formulation Aqueous Formulati ll ) ) ) ) ) )

[0661] There was no relationship between the severity of AEs and dose. Overall, there was no difference in the AE profile between the two formulations at the 20 mg dose. No consistent clinically meaningful changes were noted in the clinical laboratory results, vital signs, or ECG results.

[0662] A total of 58 samples were screened for the presence of anti-rusfertide antibodies (ADA) in human serum. Of these, all samples except one were considered negative. One sample at the early termination visit from one subject, a 57-year-old male who discontinued from the study after the second treatment, yielded an optical density value above the corresponding plate specific cut point (PSCP) with a positive titer of 507 units. Evaluation of the rusfertide concentrations in this subject did not indicate any difference in the exposure compared to other subjects. Apart from injection site erythema following a 60 mg dose that led to treatment discontinuation, the subject also did not report any other TEAEs.Attorney Docket No. T0947.70006WO00 Discussion

[0663] Rusfertide is a peptide mimetic of the natural peptide hormone hepcidin that is currently under investigation for the treatment of PV. Clinical trials in healthy subjects and phase 2 trials in patients with PV or hemochromatosis used rusfertide as an aqueous formulation. In phase 2 trials in patients with PV, the aqueous formulation demonstrated significant efficacy in maintaining hematocrit <45%, essentially eliminating the need for phlebotomies, and demonstrated long-term control of hematocrit. While the aqueous formulation was suitable for phase 2 studies, this formulation was not considered suitable for phase 3 and long-term trials. A lyophilized formulation provides a longer shelf life allowing for global trials over a wider geographic temperature range. Lyophilization is often used as a means of improving the shelf life of peptide and protein drugs that are frequently subject to instabilities due to the presence of water. The current study investigated the dose-ranging pharmacokinetics, pharmacodynamics, and safety of a lyophilized formulation of rusfertide (10 to 60 mg) and an aqueous formulation (20 mg) in healthy volunteers. The recommended starting dose of rusfertide for both formulations is 20 mg, supporting the use of this dose strength for comparison of the two formulations. In clinical practice, the dose of rusfertide is titrated for each subject to identify a dose that maintains hematocrit levels below 45%. The mean weekly rusfertide dose in subjects who completed the open-label dose-titration phase of the phase 2, REVIVE study was 41.3 mg, supporting the dose range of 10 to 60 mg selected for the current study. Since the dose of rusfertide is titrated to effect, the current pharmacokinetic and pharmacodynamic investigation was not designed as a bioequivalence trial and rather was intended to provide an understanding of the improvement in the rusfertide exposure from the lyophilized formulation relative to the previously studied aqueous formulation.

[0664] Following SC administration, rusfertide plasma concentrations were noted within 1 hour, the first sampling time point. Median peak concentrations occurred 24 to 48 hours following dose administration for the 10 to 30 mg doses of the lyophilized formulation and occurred earlier for the 45 and 60 mg doses. The SC absorption of rusfertide is extended, possibly a result of dual absorption pathways, which in combination with a modestly long elimination half-life results in sustained plasma concentrations and a delayed Tmax. An earlier Tmax is noted at higher doses, possibly reflecting an increase in the fraction of rusfertide absorbed rapidly compared to a portion that may be absorbed more slowly at lower doses.Attorney Docket No. T0947.70006WO00

[0665] Similar to a previous study with the aqueous formulation, a less-than-proportional increase in exposure (Cmax and AUC) was seen for the lyophilized formulation over the dose range of 10 to 60 mg in the current study. The mean elimination half-life was 19.6 to 57.1 hours. There was a trend of increasing CL / F and Vz / F with increasing rusfertide dose, which may explain the less than proportional increases in Cmax and AUC. Rusfertide likely follows clearance through linear and nonlinear (saturable, potentially target-mediated drug disposition) mechanisms. The increase in apparent clearance and volume of distribution with increasing dose may be reflective of the saturation of the nonlinear clearance, resulting in an increase in the amount of free drug and a higher rate of clearance. This nonlinear clearance may be reflective of target mediated drug disposition, which would be consistent with the mechanism of action of rusfertide with saturable binding to the ferroportin receptor.

[0666] Compared with the aqueous formulation, lyophilized rusfertide had an approximately 1.5- fold higher AUC at 20 mg. The pharmacokinetics of the 20 mg dose of the rusfertide aqueous formulation was comparable to that noted in a previous study. Although the lyophilized formulation resulted in higher Cmax and AUC values compared to the aqueous formulation at 20 mg, there did not appear to be a difference in the pharmacodynamic effect on serum iron and TSAT at this dose level between the two formulations. We attribute this partly to the asymptotic concentration-effect relationship wherein the effect on change in serum iron and TSAT is similar for the two formulations up to approximately a rusfertide concentration of 130 ng / mL, the common range in rusfertide concentrations, above which there is a relatively smaller pharmacodynamic effect, with further increases in rusfertide concentration (Figure 16).

[0667] M1 and M6 comprise <1% of the total drug-related AUC and are considered minor metabolites. According to the metabolites in safety testing (MIST) guidelines, M9, and conservatively M4, are considered major metabolites for the lyophilized formulation since their AUC values comprise >10% of the total AUC. At the 20 mg dose, rusfertide AUCinf comprised 65.5% of the total drug-related AUCinf for the lyophilized formulation and 53.7% for the aqueous formulation. M4 comprised 8.4% of the total drug-related AUC for the lyophilized formulation and 20.9% for the aqueous formulation. In contrast, there was a smaller difference in the M9 exposure between the two formulations; M9 comprised 29.7% of the total AUC for the lyophilized formulation and 24.9% for the aqueous formulation.Attorney Docket No. T0947.70006WO00

[0668] Median peak concentrations for both major metabolites, M4 and M9, occurred similar to or later than that of rusfertide, suggesting the absence of presystemic metabolism for these metabolites. The major metabolites, M4 and M9, had similar mean elimination half-lives as rusfertide, suggesting that these two metabolites follow formation rate limited pharmacokinetics.

[0669] There is increasing recent interest in the use of hepcidin, mini-hepcidins, and targeting transmembrane serine protease 6 (TMPRSS6) using antisense nucleotides, small interfering RNA (siRNA) or monoclonal antibodies for iron overload disorders. Hepcidin is reported to have a rapid clearance, primarily through renal excretion and reabsorption which limits it use as a therapeutic. Rusfertide appears to have favorable PK and PD characteristics compared with some other investigational agents that have effects on the ferroportin receptor.

[0670] SC administration of LJPC-401, a synthetic hepcidin, in healthy subjects resulted in peak concentrations at approximately 2 hours and the mean terminal half-life ranged from ~3 to 11 hours. Mean maximum reduction in serum iron with LJPC-401 occurred 4- to 8 hours postdose with a return to baseline within 48 hours. In contrast, SC rusfertide as the lyophilized formulation has a more sustained absorption, with peak concentrations noted approximately 4- to 24 hours following injection and a longer apparent elimination half-life, supporting less frequent dosing. Consistent with the pharmacokinetic profile, rusfertide led to dose-related, rapid, robust, and sustained effects on serum iron and transferrin-iron saturation, with effects noted within 1 hour of dose administration. Maximum reductions in serum iron and TSAT were noted approximately 24 to 48 hours following rusfertide administration with subsequent dose-dependent return to baseline generally at 168 hours or later.

[0671] VIT-2763, a small-molecule oral ferroportin inhibitor, has a median Tmax of 0.5 to 3 hours and a geometric mean elimination half-life of 1.9 to 5.3 hours following single doses in healthy subjects

[0044] . Correspondingly, the nadir in serum iron levels was observed 4 to 8 hours post-dose, and mean serum iron levels rebounded to baseline or above by 24 hours post-dose. The longer duration of effect with rusfertide (nadir at 24 hours which is sustained until 72 to 96 hours) would allow once- or twice-weekly dosing.

[0672] This pharmacokinetic and pharmacodynamic trial in healthy subjects guided the choice of the starting dose of 20 mg for the planned Phase 3 safety and efficacy study and ensured patient safety given the demonstrated tolerability of the range of rusfertide exposures previously studied. The similar relationship between rusfertide exposure (AUC) and serum iron AUEC (Figure 15) for bothAttorney Docket No. T0947.70006WO00 formulations suggests that given a desired reduction in serum iron, it would be possible to select the dose of rusfertide required with each formulation.

[0673] In phase 2 studies, subcutaneous rusfertide of the aqueous formulation has shown a rapid, robust, and sustained reduction in hematocrit in patients with PV, essentially eliminating the need for phlebotomies, an outcome of restricting iron availability for erythropoiesis. These findings in patients with PV are consistent with the pharmacokinetic and pharmacodynamic profile of rusfertide observed in healthy subjects. Rusfertide shows acute reductions in serum iron (Figure 13A) and an exposure-related decrease in AUEC for serum iron (Figure 15A), reflecting limited availability of serum iron for erythropoiesis.

[0674] Single doses of SC rusfertide doses were generally well tolerated. Approximately 28% of subjects experienced injection site reactions, with two subjects discontinuing the study early because of these events. No serious AEs were identified in this dose-ranging trial, and no clinically meaningful effects were seen on clinical laboratory parameters, vital signs, or ECGs. The overall safety profile of SC rusfertide was as expected for a hepcidin mimetic intended to limit iron availability.

[0675] A limitation of the current study, the presence of low detectable rusfertide concentrations in some subjects in subsequent treatment periods, was likely a result of a short (2 week) washout period. These instances were generally limited to cases where the previous treatment was 60 mg and baseline concentration generally comprised less than 7% of the peak plasma concentration on average. In these cases, the rusfertide plasma concentrations were corrected using an average elimination rate from a previous study with intravenous rusfertide. Example 16.

[0676] Background

[0677] Polycythemia vera (PV) is a chronic myeloproliferative neoplasm that is characterized by the overproduction of red blood cells (RBCs). Rusfertide is a novel, first-in-class subcutaneous (SC) injectable peptide mimetic of hepcidin, the principal regulator of iron homeostasis. Like hepcidin, rusfertide binds to ferroportin, a transmembrane channel that regulates iron transport and controls iron availability for RBC production in the bone marrow. In the phase 2 REVIVE study (NCT04057040) that investigated rusfertide in patients with PV, the primary endpoint for response was met (responders defined as patients with hematocrit (Hct) control, no therapeutic phlebotomyAttorney Docket No. T0947.70006WO00 (TP) use, and completion of the 12-week randomized, placebo-controlled regimen during Part 2 (Kremyanskaya M, et al. N Engl J Med.2024;390(8):723-35).

[0678] Aims

[0679] The phase 3 VERIFY study (NCT05210790) was designed to assess the efficacy and safety of rusfertide in TP-dependent patients with PV who are receiving standard-of-care therapy.

[0680] VERIFY is an ongoing, global, phase 3 randomized three-part study evaluating SC rusfertide in adult patients with PV who require frequent TP (defined as ≥3 TP in 28 weeks or ≥5 TP in 52 weeks prior to enrollment) with or without cytoreductive therapy (CRT) to maintain Hct <45%. During Part 1a (double-blind period), approximately 250 patients (293 patients) were randomized (1:1) to receive rusfertide (starting dose: 20 mg once weekly (QW)) or placebo that was added to patients’ ongoing therapy (ie, TP with or without CRT) for 32 weeks. Patients were stratified by baseline therapy. All patients who complete Part 1a are eligible to participate in Part 1b (open-label treatment period, Weeks 32-52), including patients who receive placebo during Part 1a (i.e., who can cross over to receive rusfertide). All patients who complete Part 1b are eligible to continue receiving open label rusfertide in Part 2 (long-term extension period, Weeks 52-156). The primary efficacy endpoint is the proportion of patients achieving a response (defined as the absence of TP eligibility, i.e., confirmed Hct ≥45% and ≥3% higher than baseline Hct or Hct ≥48%) from Weeks 20-32 (inclusive). Key secondary endpoints assessed from Weeks 0-32 (inclusive) include a comparison of rusfertide to placebo for the mean number of TPs and the proportion of patients with Hct <45%. Other key secondary endpoints include comparison of rusfertide to placebo for the mean change from baseline at the end of Part 1a (Week 32) in the Patient-Reported Outcomes Measurement Information System (PROMIS) Fatigue Short Form 8a total T-score and in the Myelofibrosis Symptom Assessment Form version 4.0 (MFSAF) Total Symptom Score (TSS7). There is a 32- week period during which rusfertide or placebo will be added-on to each subject's ongoing therapy for polycythemia vera which may include phlebotomy only or phlebotomy plus stable doses of either of hydroxyurea, interferon and / or ruxolitinib. Approximately 6 and 12 months after their last dose of rusfertide, subjects will have a post-study contact (e.g. by phone) for safety.

[0681] Eligibility Criteria

[0682] Main inclusion criteria: All subjects must meet all of the following inclusion criteria to be enrolled.Attorney Docket No. T0947.70006WO00 • Male and female subjects aged 18 (or the country specific minimum age of consent > 18) years or older. • Meet revised 2016 World Health Organization (WHO) criteria for the diagnosis of polycythemia vera. • At least 3 therapeutic phlebotomies (TPs) due to inadequate hematocrit control in 6 months before randomization or at least 5 TPs due to inadequate hematocrit control in 1 year before randomization. • Complete blood count (CBC) values immediately prior to randomization: o Hematocrit < 45%, o White blood cell (WBC) of 4,000 / µL to 20,000 / µL (inclusive), and o Platelets 100,000 / µL to 1,000,000 / µL (inclusive). • Subjects receiving cytoreductive therapy at randomization must be on a stable PV therapy regimen. • Subjects treated with TP alone at randomization must have stopped cytoreductive therapy 2 to 6 months before screening.

[0683] Main exclusion criteria: Subjects must meet none of the following exclusion criteria to be enrolled. • Clinically meaningful laboratory abnormalities at Screening. • Subjects who require TP at hematocrit levels lower than 45%. • Clinically significant thrombosis (e.g., deep vein thrombosis or splenic vein thrombosis) within 2 months prior to randomization. • Active or chronic bleeding within 2 months prior to randomization. • History of invasive malignancies within the last 5 years, except localized cured prostate cancer and cervical cancer. • Subjects with in situ or stage 1 squamous cell carcinoma of the skin, in situ or stage 1 basal cell carcinoma of the skin, or in situ melanoma of the skin identified during screen unless the cancer is adequately treated before randomization. • Received Busulfan, Pipobroman or 32Phosphorus within 7 months prior to screening.

[0684] Interventions

[0685] Part 1a: Randomized, double-blindAttorney Docket No. T0947.70006WO00

[0686] Subjects will be randomized in a blinded fashion to 32 weeks of rusfertide or placebo added- on to each subject’s ongoing treatment for polycythemia vera (PV)

[0687] Part 1b and Part 2: Open-label

[0688] Open-label treatment phase during which all subjects who complete Part 1a successfully will receive rusfertide for 124 weeks.

[0689] Outcome Measures

[0690] Primary Outcome Measure

[0691] Proportion of subjects achieving a response who receive rusfertide compared to placebo. Response is defined as the absence of phlebotomy eligibility. The time frame for primary outcome measure is week 20 through week 32.

[0692] Secondary Outcome Measures

[0693] Numerous secondary outcome measures including: (1) comparison of mean number of phlebotomies between rusfertide and placebo (week 0 compared to week 32); (2) proportion of subjects with HCT values <45% for rusfertide and placebo (week 0 and week 32); (3) comparison of mean change from baseline in total fatigue score based on PROMIS Short Form between rusfertide and placebo (week 32); and (4) comparison of mean change from baseline in total MFSAF total score (week 32).

[0694] Results A total of 293 patients (median age, 57 years) were randomized to receive rusfertide (n=147) or placebo (“PBO,” n=146). In the rusfertide and PBO groups, 56.5% (n=83) and 55.5% (n=81) patients (“pts”), respectively, received concurrent cytoreductive therapy (CRT). During weeks 20- 32, significantly more patients in the rusfertide group (76.9%) achieved a clinical response versus patients in the PBO group (32.9%) (p<0.0001). The least squares means (“LS means”) number of phlebotomies (“PHLs”) (Weeks 0-32) was 0.5 (0.2) in the rusfertide group versus 1.8 (0.2) in the PBO group (<0.0001). More than four times as many rusfertide patients had hematocrit (“Hct”) <45% vs PBO patients (62.6% [n=92] and 14.4% [n=21], respectively; p<0.0001). For the PROMIS Fatigue SF-8a total T-score and the MFSAF v4.0 , the least-squares mean difference (rusfertide – PBO) was statistically significant in favor of rusfertide for both endpoints (p<0.03). During Part 1a, the most common treatment-emergent adverse events (AEs) in the rusfertide and PBO groups, respectively, were injection site reactions (55.9% and 32.9%), anemia (15.9% and 4.1%), and fatigue (15.2% and 15.8%). Serious AEs were reported 3.4% and 4.8% in the rusfertide and PBO groups,Attorney Docket No. T0947.70006WO00 respectively; none were considered to be related to rusfertide. During Part 1a, cancer events were reported in one patient (0.7%) in the rusfertide group and 7 pts (4.8%) in the PBO group.

[0695] Conclusions

[0696] In patients with PV receiving standard of care (“SOC”), rusfertide resulted in a statistically significant reduction in the mean number of PHLs, increase in Hct control, and improvement in symptoms (measured by the PROMIS Fatigue SF-8a and MSFAF v4.0) vs. PBO. Rusfertide is the first investigational therapy to prospectively demonstrate a statistically significant improvement in patient-reported outcomes in patients with PV. Rusfertide had a safety and tolerability profile consistent with rusfertide in prior PV studies, including the REVIVE phase 2 study.

Claims

Attorney Docket No. T0947.70006WO00 CLAIMS What is claimed is:

1. A pharmaceutical composition comprising: (i) a peptide comprising an amino acid sequence: Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2[SEQ ID NO: 1] or a pharmaceutically acceptable salt or solvate thereof, and (ii) a zinc salt, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:

1.

2. The pharmaceutical composition of claim 1, wherein the peptide comprises a disulfide bond between two cysteine residues of the peptide.

3. The pharmaceutical composition of claim 2, wherein the peptide comprises a structure of formula (I): .

4. The pharmaceutical composition of claim 1, wherein the molar ratio of the zinc salt and the peptide is from about 0.2:1 to about 2.5:1.Attorney Docket No. T0947.70006WO00 5. The pharmaceutical composition of claim 1, wherein the molar ratio of the zinc salt and the peptide is from about 0.3:1 to about 1.2:

1.

6. The pharmaceutical composition of claim 1, wherein the molar ratio of the zinc salt and the peptide is from about 0.75:1 to about 1:

1.

7. The pharmaceutical composition of claim 1, wherein the zinc salt is zinc maleate, zinc tartrate, zinc oxide, zinc hydroxide, zinc mesylate, zinc besylate, zinc chloride or zinc acetate.

8. The pharmaceutical composition of any one of claims 1-7, wherein the zinc salt is zinc acetate.

9. The pharmaceutical composition of any one of claims 1-7, wherein the zinc salt is zinc chloride.

10. The pharmaceutical composition of any one of claims 1-9, wherein the composition is a liquid formulation.

11. The pharmaceutical composition of any one of claims 1-10, wherein the composition is a reconstituted lyophilized formulation.

12. The pharmaceutical composition of any one of claims 1-10, wherein the composition maintains a purity of greater than 97% at room temperature for at least 12 months.

13. The pharmaceutical composition of any one of claims 1-10, wherein the composition maintains a purity of greater than 95% at room temperature for at least 24 months.

14. The pharmaceutical composition of any one of claims 1-10, further comprising a tonicity adjusting agent, a buffering agent, and a stabilizer.

15. The pharmaceutical composition of claim 11, further comprising a buffering agent and a bulking agent.

16. The pharmaceutical composition of any one of claims 1-10, wherein the composition is a solution comprising one or more solvents.Attorney Docket No. T0947.70006WO00 17. The pharmaceutical composition of claim 16, wherein the solvent is water, dimethylsulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide or a combination thereof.

18. The pharmaceutical composition of any one of claims 1-17, wherein the peptide in the composition is present in an amount from about 0.1% w / v to about 15% w / v.

19. The pharmaceutical composition of claim 10, wherein the peptide in the composition is from about 1% w / v to about 6% w / v.

20. The pharmaceutical composition of claim 11, wherein the peptide in the composition is from about 2% w / w to about 12% w / w.

21. The pharmaceutical composition of claim 1, wherein the dosage of the peptide is from about 1 mg to about 100 mg.

22. The pharmaceutical composition of claim 1, wherein the dosage of the peptide is about 10 mg, about 20 mg, about 30 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg or about 100 mg.

23. A liquid pharmaceutical composition comprising a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, a buffering agent, a zinc salt, a stabilizer, a tonicity adjuster, a pH adjuster, and a solvent, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:

1.

24. The liquid pharmaceutical composition of claim 23, wherein the buffering agent comprises sodium acetate, wherein the zinc salt comprises zinc chloride, wherein the stabilizer comprises PEG 3350, wherein the tonicity adjuster comprises sorbitol, wherein the pH adjuster comprises sodium hydroxide and acetic acid, and wherein the solvent comprises water.

25. The liquid pharmaceutical composition of claim 23, wherein the dosage of the peptide is about 10 mg, about 20 mg, about 30 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg or about 100 mg.Attorney Docket No. T0947.70006WO00 26. The liquid pharmaceutical composition of any one of claims 23-25, wherein the composition has purity greater than 97% after 12 months.

27. The liquid pharmaceutical composition of any one of claims 23-25, wherein the composition has less than about 2.0 % Asp-related degradants after 24 months.

28. A liquid pharmaceutical composition comprising: 0.1% - 12% (w / v) of a peptide comprising SEQ ID NO: 1; 0% - 0.68% (w / v) sodium acetate trihydrate; 0.0004% - 2.0% (w / v) zinc chloride; 0% - 5.5% (w / v) sorbitol; 0% - 3.6% PEG3350; sodium hydroxide or acetic acid as needed to pH 5.6; and water as needed so that the total volume of the liquid pharmaceutical composition is 1.0 mL.

29. A liquid pharmaceutical composition comprising: 0.2% - 4.0% (w / v) of a peptide comprising SEQ ID NO: 1; 0.272% (w / v) sodium acetate trihydrate; 0.01% - 0.45% (w / v) zinc chloride; 3.1% - 5.5% (w / v) sorbitol; 1.8% (w / v) PEG 3350; sodium hydroxide or acetic acid as needed to pH 5.6; and water as needed so that the total volume of the liquid pharmaceutical composition is 1.0 mL.

30. A liquid pharmaceutical composition comprising: 1% - 6% (w / v) of a peptide comprising SEQ ID NO: 1; 0.9% - 4.9% (w / v) sorbitol; 0.272% (w / v) sodium acetate trihydrate; 0.11% - 0.70% (w / v) zinc chloride; 1.8% (w / v) PEG 3350; glacial acetic acid as needed to pH 5.4; and water as needed so that the total volume of the liquid pharmaceutical composition is 0.5 mL - 1.0 mL.Attorney Docket No. T0947.70006WO00 31. A reconstituted lyophilized pharmaceutical composition comprising a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, a bulking agent, and a zinc salt, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:

1.

32. The reconstituted lyophilized pharmaceutical composition of claim 31, wherein the bulking agent is mannitol, and wherein the zinc salt comprises zinc acetate.

33. The reconstituted lyophilized pharmaceutical composition of claim 31, wherein the composition is reconstituted for injection and comprises: 2% - 12% (w / v) of a peptide of SEQ ID NO: 1; 4% - 5% (w / v) mannitol; 0.3% (w / v) sodium acetate trihydrate; 0.2% (w / v) zinc acetate; glacial acetic acid as needed to pH 5.4; and water as needed so that the total volume of the liquid pharmaceutical composition is 0.5 mL.

34. The reconstituted lyophilized pharmaceutical composition of claim 31, wherein the dosage of the peptide is about 10 mg, about 20 mg, about 30 mg, about 45 mg or about 60 mg.

35. A kit for subcutaneous injection comprising: a. a lyophilized pharmaceutical composition comprising a peptide having an amino acid sequence of Isovaleric acid-DTHFPCI(K(isoGlu-Palm))FEPRSKGCK-NH2 (SEQ ID NO: 1) or a pharmaceutically acceptable salt or solvate thereof, and b. a reconstitution liquid comprising a zinc salt, wherein the molar ratio of the zinc salt and the peptide is from about 0.1:1 to about 3:

1.

36. A method for treating a disease of iron metabolism in a subject, the method comprising administering to the subject a therapeutically effective amount of a liquid pharmaceutical composition of any one of claims 1-30 or a reconstituted lyophilized pharmaceutical composition of any one of claims 31-34.

37. The method of claim 36, wherein the disease of iron metabolism is an iron overload disease.Attorney Docket No. T0947.70006WO00 38. A method for treating polycythemia vera in a subject in need thereof, the method comprising administering to the subject in need thereof a therapeutically effective amount of a liquid pharmaceutical composition of any one of claims 1-30 or a reconstituted lyophilized pharmaceutical composition of any one of claims 31-34.

39. The method of claim 38, wherein the administration is subcutaneous injection.

40. The method of claim 39, wherein the subcutaneous injection is administered using an autoinjector.

41. The method of any one of claims 38-40, wherein the composition is administered at a rate to achieve: a. an average peptide plasma area under the curve (AUC) of at least 32000 h*ng / mL for a composition comprising about 20 mg, 30 mg, 45 mg or 60 mg of Compound 1; and b. an average maximum peptide blood plasma concentration (average Cmax) of at least 500 ng / mL per each dosage of 20 mg, 30 mg, 45 mg or 60 mg of Compound 1 delivered; wherein the AUC is measured from time zero to the time of last measured concentration.

42. The method of any one of claims 38-41, wherein the zinc salt in the liquid pharmaceutical composition or the reconstituted lyophilized composition is zinc chloride.

43. The method of any one of claims 38-42, wherein the zinc salt and the peptide in the liquid pharmaceutical composition or the reconstituted lyophilized composition have a molar ratio of 0.2:1, 1:1, or 2:

1.

44. The method of any one of claims 38-43, wherein the peptide of SEQ ID NO: 1 is administered once weekly or twice weekly.

45. The method of any one of claims 38-44, wherein the peptide in the composition is present in an amount from about 0.1% w / v to about 15% w / v.Attorney Docket No. T0947.70006WO00 46. The method of any one of claims 38-44, wherein the peptide in the composition is present in an amount from about 1% w / v to about 6% w / v.

47. The method of any one of claims 38-44, wherein the peptide in the composition is present in an amount from about 2% w / w to about 12% w / w.

48. The method of any one of claims 38-44, wherein the dosage of the peptide is from about 1 mg to about 100 mg.

49. The method of any one of claims 38-44, wherein the dosage of the peptide is about 10 mg, about 20 mg, about 30 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg or about 100 mg.

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