FORMULATION OF MK2 INHIBITOR PEPTIDES
The described invention provides pharmaceutical formulations comprising a polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or a functional equivalent thereof with improved stability and bioavailability.
1 . A pharmaceutical formulation for delivery by inhalation comprising a therapeutic amount of an MK2i polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or a functional equivalent thereof, up to 5% w/w solids before drying, up to 7.3% glycerin, and a pharmaceutically acceptable carrier, wherein the formulation:
(a) is isosmotic and non-buffered;
(b) has a stable pH of about 7.0 for at least 2 weeks at 60° C.; and
(c) is effective to preserve at least about 93% stability of physical, chemical, microbiological, therapeutic, and toxicological specifications of the MK2i polypeptide for at least 2 weeks at 60° C., and bioavailability of the MK2i polypeptide.
2 . The pharmaceutical formulation according to claim 1 , wherein the pharmaceutical formulation is a particulate pharmaceutical formulation.
3 . The pharmaceutical formulation according to claim 1 , wherein the pharmaceutical formulation is an aerosolized pharmaceutical formulation.
4 . The pharmaceutical formulation according to claim 1 , wherein the formulation is prepared by a process of spray drying.
5 . The pharmaceutical formulation according to claim 2 , wherein the pharmaceutical formulation comprises 1% w/w solids before drying.
6 . The pharmaceutical formulation according to claim 2 , wherein the pharmaceutical formulation comprises 5% w/w solids before drying.
7 . The pharmaceutical formulation according to claim 1 further comprising trehalose.
8 . The pharmaceutical formulation according to claim 1 , wherein the functional equivalent is made from a fusion between a first polypeptide that is a protein transduction domain (PTD) and a second polypeptide that is a therapeutic domain (TD).
9 . The pharmaceutical formulation according to claim 8 , wherein the protein transduction domain (PTD) is selected from the group consisting of a polypeptide of amino acid sequence YARAAARQARA (SEQ ID NO: 11), FAKLAARLYR (SEQ ID NO: 16), and KAFAKLAARLYR (SEQ ID NO: 17), and a second polypeptide that is a therapeutic domain (TD) of amino acid sequence KALARQLGVAA (SEQ ID NO: 2).
10 . The pharmaceutical formulation according to claim 1 , wherein the pharmaceutical formulation is delivered to a subject via a dry powder inhalation device (DPI).
11 . The pharmaceutical formulation according to claim 3 further comprising saline before drying.
12 . The pharmaceutical formulation according to claim 11 , wherein the saline is NaCl.
13 . The pharmaceutical formulation according to claim 12 , wherein the polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or the functional equivalent thereof is at a concentration of 0.7 mg/mL.
14 . The pharmaceutical formulation according to claim 12 , wherein the polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or the functional equivalent thereof is at a concentration of 7.0 mg/mL.
15 . The pharmaceutical formulation according to claim 3 , wherein the pharmaceutical formulation is formulated to be used via a nebulizer.
16 . The pharmaceutical formulation according to claim 1 , comprising an ionic complex of the MK2i polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or a functional equivalent thereof and a nano-polyplex polymer, wherein the ionic complex dissociates in intracellular compartments selected by intracellular pH conditions such that bioactivity and stability of the peptide is preserved.
17 . The pharmaceutical formulation according to claim 16 , wherein the nano-polyplex polymer is anionic and endosomolytic.
18 . The pharmaceutical formulation according to claim 17 , wherein the nano-polyplex polymer is poly(propylacrylic acid) (PPAA).
19 . The pharmaceutical formulation according to claim 16 , wherein the pharmaceutical formulation comprises a charge ratio (CR) of the MK2i polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or a functional equivalent thereof to PPAA selected from the group consisting of 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:1.5, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 and 1:10.
20 . The pharmaceutical formulation according to claim 19 , wherein the charge ratio (CR) is 1:3.
21 . The pharmaceutical formulation according to claim 16 , wherein the functional equivalent is made from a fusion between a first polypeptide that is a protein transduction domain (PTD) and a second polypeptide that is a therapeutic domain (TD).
22 . The pharmaceutical formulation according to claim 21 , wherein the protein transduction domain (PTD) is selected from the group consisting of a polypeptide of amino acid sequence YARAAARQARA (SEQ ID NO: 11), FAKLAARLYR (SEQ ID NO: 16), and KAFAKLAARLYR (SEQ ID NO: 17), and a second polypeptide that is a therapeutic domain (TD) of amino acid sequence KALARQLGVAA (SEQ ID NO: 2).
23 . The pharmaceutical formulation according to claim 16 , wherein the pharmaceutical formulation is delivered to a subject via an implantation device.
24 . The pharmaceutical formulation according to claim 16 , wherein the pharmaceutical formulation is delivered to a subject topically.
25 . The pharmaceutical formulation according to claim 16 , wherein the pharmaceutical formulation is delivered to a subject parenterally.
26 . A method for treating a vascular graft-induced intimal hyperplasia in a subject in need of such treatment, the method comprising administering the pharmaceutical formulation of claim 16 comprising a therapeutic amount of a polypeptide of amino sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or a functional equivalent thereof, and a nano-polyplex polymer, wherein the therapeutic amount is effective to inhibit MK2; and to treat a vascular graft-induced intimal hyperplasia.
27 . The method according to claim 26 , wherein the nano-polyplex polymer is anionic and endosomolytic.
28 . The method according to claim 27 , wherein the nano-polyplex polymer is poly(propylacrylic acid) (PPAA).
29 . The method according to claim 26 , wherein the pharmaceutical formulation comprises a charge ratio (CR) of the polypeptide of amino acid sequence YARAAARQARAKALARQLGVAA (SEQ ID NO: 1) or a functional equivalent thereof to PPAA selected from the group consisting of 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:1.5, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 and 1:10.
30 . The method according to claim 29 , wherein the charge ratio (CR) is 1:3.
31 . The method according to claim 29 , wherein the nano-polyplex polymer is poly(acrylic acid) (PAA).
32 . The method according to claim 26 , wherein the functional equivalent is made from a fusion between a first polypeptide that is a protein transduction domain (PTD) and a second polypeptide that is a therapeutic domain (TD).
33 . The method according to claim 32 , wherein the protein transduction domain (PTD) is selected from the group consisting of a polypeptide of amino acid sequence YARAAARQARA (SEQ ID NO: 11), FAKLAARLYR (SEQ ID NO: 16), and KAFAKLAARLYR (SEQ ID NO: 17), and a second polypeptide that is a therapeutic domain (TD) of amino acid sequence KALARQLGVAA (SEQ ID NO: 2).
34 . The method according to claim 26 , wherein the administering is by an implantation device.
35 . The method according to claim 26 , wherein the administering is by topical administration.
36 . The method according to claim 26 , wherein the administering is by parenteral administration.