# US 2014/0154308 A1 · Microparticle Formulations for Delivery to the Upper and Central Respiratory Tract and Methods of Manufacture

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- Number: US 2014/0154308 A1
- Document ID: US20140154308A1
- Status: Published application
- Kind: A1
- Application number: 13/931,401
- Filed: 2013-06-28
- Published: 2014-06-05
- Earliest claimed: 2012-06-28
- Applicants: Ansun Biopharma, Inc.
- Inventors: Tiejun Li; Brian Thome
- Web page: https://patentgrep.com/patent/US20140154308A1

## CPC

- A61K 9/14: Particulate form, e.g. powders, {Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles (microspheres A61K9/16; microcapsules A61K9/50; nanocapsules, nanoparticles of the matrix type A61K9/51)}
- A61K 9/0075: {for inhalation via a dry powder inhaler [DPI], e.g. comprising micronized drug mixed with lactose carrier particles}
- A61K 9/1617: {Organic compounds, e.g. phospholipids, fats}
- A61K 9/1623: {Sugars or sugar alcohols, e.g. lactose; Derivatives thereof; Homeopathic globules}
- C07K 14/001: {by chemical synthesis}
- A61K 38/47: acting on glycosyl compounds (3.2), e.g. cellulases, lactases
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

Microparticle formulations are produced by contacting an aqueous solution of a protein or other active agent with an organic solvent, a counterion and a scavenging agent, and chilling the solution. The microparticles are useful for preparing stable, uniform pharmaceuticals of predetermined defined dimensions.

## Claims

### Claim 1

A method of making a composition comprising microparticles comprising DAS 181, the method comprising:
a) providing a feedstock composition comprising DAS181, a counterion and an organic solvent; and
b) cooling the feedstock composition to below 25° C., whereby a composition comprising microparticle comprising DAS 181 is formed.

### Claim 2 (depends on 1)

The method of claim 1 wherein the DAS181 comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:1.

### Claim 3 (depends on 1)

The method of claim 1 wherein the DAS 181 comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:2.

### Claim 4 (depends on 1)

The method of claim 1 wherein the DAS181 comprises a polypeptide consisting of the amino acid sequence of SEQ ID NO:1.

### Claim 5 (depends on 1)

The method of claim 1 wherein the DAS 181 comprises a polypeptide consisting of the amino acid sequence of SEQ ID NO:2.

### Claim 6 (depends on 1)

The method of claim 1 wherein the DAS 181 comprises a polypeptide consisting of amino acid sequence of SEQ ID NO:1 and a polypeptides consisting of the amino acid sequence of SEQ ID NO:2.

### Claim 7 (depends on 1)

The method of claim 1 wherein the feedstock composition comprises DAS 181 consisting of the amino acid sequence SEQ ID NO:1.

### Claim 8 (depends on 1)

The method of claim 1 wherein the feedstock composition comprises DAS 181 consisting of the amino acid sequence SEQ ID NO:2.

### Claim 9 (depends on 1)

The method of claim 1 wherein the feedstock composition comprises DAS 181 comprising or consisting of the amino acid sequence SEQ ID NO:1 and DAS 181 comprising or consisting of the amino acid sequence SEQ ID NO:2.

### Claim 10 (depends on 1)

The method of claim 1 where the feedstock composition comprises polypeptides consisting of the amino acid sequence SEQ ID NO:1 and polypeptides consisting of the amino acid sequence SEQ ID NO:2.

### Claim 11 (depends on 1)

The method of claim 1 wherein the step of cooling the feedstock composition includes cooling to a first temperature below 25° C. at a constant first rate and then cooling to a second temperate below the first temperature at a constant second rate that can be the same as or different from the first rate.

### Claim 12 (depends on 1)

The method of claim 1 wherein the step of cooling includes holding the feedstock composition at a first holding temperature below 25° C. for a first period of time.

### Claim 13 (depends on 12)

The method of claim 12 wherein the step of cooling includes holding the feedstock composition at a second holding temperature below the first holding temperature for a second period of time that can be the same as or different from the first period of time.

### Claim 14 (depends on 1)

The method of claim 1 wherein the step of providing a feedstock composition comprising DAS181, a counterion and an organic solvent comprises providing a solution comprising DAS 181 and a counterion in an aqueous solvent and adding an organic solvent to the solution comprising DAS 181 and a counterion in an aqueous solvent.

### Claim 15

The method of any of the forgoing claims further comprising isolating the microparticles from the composition comprising microparticles, whereby a dry powder formulation of microparticles is formed.

### Claim 16 (depends on 15)

The method of claim 15, wherein the step of isolating the microparticles comprises heating the composition comprising microparticles to a temperature and for a time that reduces the amount of organic solvent the composition comprising microparticles.

### Claim 17 (depends on 1)

The method claim 1, wherein the feedstock composition comprises two or more counterions.

### Claim 18 (depends on 1)

The method of claim 1, wherein the counterion comprises citric acid.

### Claim 19 (depends on 1)

The method of claim 1, wherein the counterion comprises magnesium sulfate.

### Claim 20 (depends on 1)

The method of claim 1 wherein the feedstock composition is 70-80% (wt/wt) DAS 181.

### Claim 21 (depends on 1)

The method of claim 1 wherein the feedstock composition is 8-12% (wt/wt) histidine and histidine HCl.

### Claim 22 (depends on 1)

The method of claim 1 wherein the feedstock composition is 8-10% (wt/wt) trehalose.

### Claim 23 (depends on 1)

The method of claim 1 wherein the feedstock composition is 5-9% (wt/wt) magnesium sulfate.

### Claim 24 (depends on 1)

The method of claim 1 wherein the feedstock composition is 1-4% (wt/wt) citrate.

### Claim 25 (depends on 1)

The method of claim 1 wherein the feedstock composition is 20-30% (wt/wt) isopropanol.

### Claim 26

The method of any of the forgoing claims, wherein the wt/wt amount of DAS181 in the resulting microparticles when substantially dry is about 60% to about 75% (wt/wt).

### Claim 27 (depends on 26)

The method of claim 26 wherein the resulting microparticles are 60% to 75% (wt/wt) a polypeptide comprising the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.

### Claim 28 (depends on 26)

The method of claim 26 wherein the resulting microparticles are 60% to 75% (wt/wt) a polypeptide consisting of comprising the amino acid sequence SEQ ID NO:1 or SEQ ID NO:2.

### Claim 29 (depends on 1)

The method of claim 1 wherein the resulting microparticles comprise a polypeptide consisting of the amino acid sequence SEQ ID NO:1 and a polypeptide consisting of comprising the amino acid sequence SEQ ID NO:2 and wherein the polypeptide consisting of SEQ ID NO:1 and the polypeptide consisting of SEQ ID NO:2 together are 60% to 75% (wt/wt) of the microparticles.

### Claim 30

The method of any of the forgoing claims, wherein the predetermined temperature below about 25° C. is between about 4° C. to about −65° C.

### Claim 31

The method any of the forgoing claims, wherein the predetermined temperature below about 25° C. is between about −20° C. to about −65° C.

### Claim 32

The method of any of the forgoing claims, wherein the predetermined temperature below about 25° C. is between about −40° C. to about −65° C.

### Claim 33

The method of any of the forgoing claims, wherein the cooling rate is about −0.1° C./min to about −1° C./min.

### Claim 34

The method of any of the forgoing claims, wherein cooling rate is about −0.1° C./min to about −0.8° C./min.

### Claim 35

The method of any of the forgoing claims, wherein cooling rate is about −0.1° C./min to about −0.7° C./min, about −0.3° C./min to about −0.5° C./min, about −0.4° C./min.

### Claim 36

The method of any of the forgoing claims, wherein the microparticles have a mass median aerodynamic diameter of between about 9 microns to about 10 microns.

### Claim 37

The method of any of the forgoing claims, further comprising adding a scavenging agent to the feedstock composition.

### Claim 38 (depends on 37)

The method of claim 37, wherein the scavenging agent is selected from among an amine, an antioxidant, a sugar, a chelator or a combination of two or more thereof.

### Claim 39 (depends on 37)

The method of claim 37, wherein the scavenging agent is histidine.

### Claim 40 (depends on 37)

The method of claim 37, wherein the scavenging agent further comprises a sugar.

### Claim 41 (depends on 40)

The method of claim 40 wherein the sugar is trehalose.

### Claim 42

The method of any of any of the forgoing claims, further comprising packaging the microparticle formulation in a container comprising between about 3 ppm to about 25 ppm aldehyde.

### Claim 43 (depends on 42)

The method of claim 42 wherein the aldehyde is formaldehyde.

### Claim 44

A composition comprising microparticles wherein the microparticles comprise about 60-70% wt/wt DAS181 (SEQ ID NO:17), 1-2% wt/wt histidine, 8-10% wt/wt histidine hydrochloride, 7-10% wt/wt trehalose, 4-8% wt/wt magnesium sulfate, 1-3% citrate, and 6-10% water.

### Claim 45 (depends on 44)

The composition of claim 44 wherein the microparticles are 60% to 75% (wt/wt) a polypeptide comprising the amino acid sequence of SEQ ID NO:1 or the amino acid sequence of SEQ ID NO:2.

### Claim 46 (depends on 44)

The composition of claim 44 wherein the microparticles are 60% to 75% (wt/wt) a polypeptide consisting of the amino acid sequence of SEQ ID NO:1 or the amino acid sequence of SEQ ID NO:2.

### Claim 47 (depends on 44)

The composition of claim 44 wherein the microparticles comprise a polypeptide consisting of the amino acid sequence of SEQ ID NO:1 and a polypeptide consisting of the amino acid sequence of SEQ ID NO:2 and wherein the polypeptide consisting of the amino acid sequence of SEQ ID NO:1 and the polypeptide consisting of the amino acid sequence of SEQ ID NO:2 together are 60% to 75% (wt/wt) of the microparticles.

### Claim 48 (depends on 44)

A composition comprising the microparticle formulation of claim 44 wherein the microparticles have an MMAD of 8-15, 8-12 or 9-10 microns.

### Claim 49 (depends on 48)

The composition of claim 48 wherein the microparticles have a GSD of 1.3-1.9, 1.4-1.8 or 1.5-1.7.

### Claim 50 (depends on 48)

The composition of claim 48 wherein the weight percent of microparticles having a aerodynamic diameter below 5 microns is less than 35%, 30%, 25%, 20%, 15%, 10%, 8%, 7%, 6%, or 5%, on a volume basis.

### Claim 51 (depends on 44)

A capsule containing a composition comprising the microparticle formulation of claim 44.

### Claim 52

A composition comprising microparticles, wherein the microparticles, when anhydrous, comprises about 65-75% wt/wt DAS181 (SEQ ID NO:1, SEQ ID NO:2 or a mixture thereof), 1-3% wt/wt histidine, 8-11% wt/wt histidine hydrochloride, 8-11% wt/wt trehalose, 5-8% wt/wt magnesium sulfate, and 1-3% citrate.

### Claim 53 (depends on 52)

The composition of claim 52 wherein the microparticles have an MMAD 8-15, 8-12 or 9-10 microns.

### Claim 54 (depends on 52)

The composition of claim 52 wherein the microparticles are 69% to 74% (wt/wt) a polypeptide comprising SEQ ID NO:1 or SEQ ID NO:2.

### Claim 55 (depends on 52)

The composition of claim 52 wherein the microparticles are 69% to 74% (wt/wt) a polypeptide consisting of SEQ ID NO:1 or SEQ ID NO:2.

### Claim 56 (depends on 52)

The composition of claim 52 wherein the microparticles comprise a polypeptide consisting of SEQ ID NO:1 and a polypeptide consisting of SEQ ID NO:2 and wherein the polypeptide consisting of SEQ ID NO:1 and the polypeptide consisting of SEQ ID NO:2 together are 66% to 74% (wt/wt) of the microparticles.

### Claim 57 (depends on 53)

The composition of claim 53 wherein the microparticles have a GSD of 1.3-1.9, 1.4-1.8 or 1.5-1.7.

### Claim 58 (depends on 57)

The composition of claim 57 wherein the weight percent of microparticles having a aerodynamic diameter below 5 microns is less than 35%, 30%, 25%, 20%, 15%, 10%, 8%, 7%, 6%, or 5%.

### Claim 59

A composition comprising microparticles wherein the microparticles comprise about 60-80% wt/wt DAS181, 4.5-6.5% wt/wt histidine, 3.5-5.5% wt/wt histidine hydrochloride, 7-11% wt/wt trehalose, 1-4% citrate, and 6-10% water.

### Claim 60 (depends on 59)

A composition of claim 59 wherein the microparticles have an MMAD of 8-15, 8-12 or 9-10 microns.

### Claim 61 (depends on 60)

The composition of claim 60 wherein the microparticles have a GSD of 1.3-1.9, 1.4-1.8 or 1.5-1.7.

### Claim 62 (depends on 59)

The composition of claim 59 wherein the microparticles are 60% to 80% (wt/wt) a polypeptide comprising SEQ ID NO:1 or SEQ ID NO:2.

### Claim 63 (depends on 59)

The composition of claim 59 wherein the microparticles are 60% to 80% (wt/wt) a polypeptide consisting of SEQ ID NO:1 or SEQ ID NO:2.

### Claim 64 (depends on 59)

The composition of claim 59 wherein the microparticles comprise a polypeptide consisting of SEQ ID NO:1 and a polypeptide consisting of SEQ ID NO:2 and wherein the polypeptide consisting of SEQ ID NO:1 and the polypeptide consisting of SEQ ID NO:2 together are 80% to 90% (wt/wt) of the microparticles.

### Claim 65 (depends on 64)

The composition of claim 64 wherein the weight percent of microparticles having an aerodynamic diameter below 5 microns is less than 35%, 30%, 25%, 20%, 15%, 10%, 8%, 7%, 6%, or 5%, on a volume basis.

### Claim 66

A composition comprising microparticles that, when anhydrous, comprise about 70-80% wt/wt DAS181, 5-7% wt/wt histidine, 4-6 wt/wt histidine hydrochloride, 9-11% wt/wt trehalose, and 1.5-3.5% citrate.

### Claim 67 (depends on 66)

The composition of claim 66, wherein the microparticles are 70 to 80% (wt/wt) a polypeptide comprising SEQ ID NO:1 or SEQ ID NO:2.

### Claim 68 (depends on 66)

The composition of claim 66, wherein the microparticles are 70% to 80% (wt/wt) a polypeptide consisting of SEQ ID NO:1 or SEQ ID NO:2.

### Claim 69 (depends on 66)

The composition of claim 66, wherein the microparticles comprise a polypeptide consisting of SEQ ID NO:1 and a polypeptide consisting of SEQ ID NO:2 and wherein the polypeptide consisting of SEQ ID NO:1 and the polypeptide consisting of SEQ ID NO:2 together are 94% to 98% (wt/wt) of the microparticles.

### Claim 70 (depends on 66)

The composition of claim 66 wherein the microparticles have an MMAD of 8-15, 8-12 or 9-10 microns.

### Claim 71 (depends on 70)

The composition of claim 70 wherein the microparticles have GSD of 1.3-1.9, 1.4-1.8 or 1.5-1.7.

### Claim 72 (depends on 70)

The composition of claim 70 wherein the weight percent of microparticles having an aerodynamic diameter below 5 microns is less than 35%, 30%, 25%, 20%, 15%, 10%, 8%, 7%, 6%, or 5%, on a volume basis.

### Claim 73 (depends on 52, 59, 66)

A capsule containing the composition of any of claim 52, 59 or 66.

### Claim 74 (depends on 73)

The capsule of claim 73 wherein the capsule contains 9-14, 10-13, 11-12, 11 or 12 mg of DAS181.

### Claim 75 (depends on 74)

The capsule of claim 74, when used in an inhaler, delivers a dose of 8-11, 9-10, 9 or 10 mg of DAS181.

### Claim 76 (depends on 75)

The capsule of claim 75, wherein the capsule further contains lactose.