Combination compositions
Combination compositions comprising self-assembling peptides and payload agents, and methods of making and using such compositions, are described.
1. A method of manufacturing a liquid combination composition comprising a self-assembling peptide and a therapeutic agent, the method comprising steps of:
before or after combining the therapeutic agent with a liquid preparation of the self-assembling peptide, homogenizing the liquid preparation by subjecting it to high shear stress sufficient to reduce its initial storage modulus by at least a factor of three;
wherein the self-assembling peptide comprises from about 7 to about 32 alternating hydrophobic and hydrophilic amino acids.
2. The method of claim 1 , wherein the initial storage modulus is about 100 Pa to about 3000 Pa at 5 rad/sec of frequency and 0.1 Pa of oscillation stress.
3. The method of claim 2 , wherein the step of homogenizing comprises subjecting to high shear stress sufficient to reduce the initial storage modulus by at least 80-400 fold.
4. The method of claim 2 , wherein the step of homogenizing comprises subjecting the liquid preparation to high shear stress by passing it through at least one shear-thinning unit.
5. The method of claim 4 , wherein the at least one shear-thinning unit is or comprises at least one needle.
6. The method of claim 5 , wherein the at least one needle is at least 1 mm long.
7. The method of claim 5 , wherein the at least one needle has a gauge greater than about 27.
8. The method of claim 4 , wherein the at least one shear-thinning unit is or comprises at least one screen with micro- or nano-sized holes.
9. The method of claim 8 , wherein the micro- or nano-sized holes have a largest dimension smaller than about 43 μm.
10. The method of claim 8 , wherein the screen is made at least in part of a material selected from the group consisting of stainless-steel, tungsten, titanium, silicon, ceramic, plastic, and combinations thereof.
11. The method of claim 8 , wherein thickness of the screen is about 10 μm to about 10 mm.
12. The method of claim 1 , wherein the high shear stress is within a range of about 30 to about 200 Pa.
13. The method of claim 1 , wherein the self-assembling peptide has an amino acid sequence selected from the group consisting of RADA16 as set forth in SEQ ID NO:1, IEIK13 as set forth in SEQ ID NO:3, and KLD12 as set forth in SEQ ID NO:2.
14. The method of claim 13 , wherein the self-assembling peptide is present at a concentration of at least 2.5% weight to volume.
15. A combination composition prepared using the method of claim 1 .
16. The method of claim 8 , wherein the micro- or nano-sized holes have a largest dimension of about 25 μm or less.
17. The method of claim 1 , wherein, prior to the step of homogenizing, the liquid preparation of the self-assembling peptide had been stored for a period of at least 2 days.
18. A method comprising steps of:
combining a liquid preparation comprising at least 1% weight/volume of a self-assembling peptide having an amino acid sequence selected from the group consisting of RADA16 as set forth in SEQ ID NO:1, IEIK13 as set forth in SEQ ID NO:3, and KLD12 as set forth in SEQ ID NO:2 with a therapeutic agent so that a combination liquid composition is created; and
before or after the step of combining, treating the liquid preparation by subjecting it to high shear stress sufficient to reduce its initial storage modulus by at least a factor of three.
19. The method of claim 18 , wherein, prior to the step of treating, the liquid preparation had been stored for a period of at least 2 days.
20. The method of claim 18 or claim 19 , wherein the liquid preparation comprises at least 2.5% by weight/volume of the self-assembling peptide.
21. The method of claim 18 , wherein, after the step of treating, the combination liquid composition is characterized by microscopic homogeneity.
22. The method of claim 2 , wherein the step of homogenizing comprises subjecting to high shear stress by one or more syringes.
23. The method of claim 18 or 19 , wherein the step of subjecting to high shear stress comprises passing the liquid preparation through at least one shear-thinning unit selected from the group consisting of needles, membranes, screens, and combinations thereof.
24. The method of claim 18 or 19 , wherein the step of subjecting to high shear stress comprises at least one step of passing from a first syringe, through a chamber with pores, into a second syringe.
25. The method of claim 24 , wherein the pores have a size pore size of about 0.45 μm to 200 μm.
26. The method of claim 25 , wherein the pore size is about 1 μm to 150 μm.
27. The method of claim 25 , wherein the pore size is about 1 μm to 100 μm.
28. The method of claim 25 , wherein the pore size is about 5 μm to 50 μm.
29. The method of claim 24 , wherein the step of passing from a first syringe, through a chamber with pores, into a second syringe comprises steps of:
providing the liquid preparation loaded into the first syringe;
pushing the liquid preparation out of the first syringe, through a connector that is or comprises the chamber with pores, and into the second syringe, loaded with the therapeutic agent so that the liquid preparation and therapeutic agent are mixed;
pushing the mixture back through the connector into the first syringe;
optionally further pushing the mixture back and/or forth through the connector between the first and second syringes.