U.S. patent application number 16/757195 was filed with the patent office on 2021-04-29 for silk fibroin-containing composition and methods of use thereof.
The applicant listed for this patent is Georgetown University. Invention is credited to Morarji PEESAY.
Application Number | 20210121525 16/757195 |
Document ID | / |
Family ID | 1000005384180 |
Filed Date | 2021-04-29 |
United States Patent
Application |
20210121525 |
Kind Code |
A1 |
PEESAY; Morarji |
April 29, 2021 |
SILK FIBROIN-CONTAINING COMPOSITION AND METHODS OF USE THEREOF
Abstract
Provided are compositions comprising silk fibroin,
perfluorocarbon (PFC), and surfactant, and methods of use thereof.
The compositions can further comprise a drug, an antibody, or a
vaccine. Compositions of the invention are useful in the treatment
of certain lung diseases and conditions, including in particular
those characterized by surfactant deficiency. Compositions of the
invention are particularly useful in the treatment of respiratory
distress syndrome (RDS). Also provided are methods for making the
compositions, and kits comprising components of the
compositions.
Inventors: |
PEESAY; Morarji;
(Germantown, MD) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Georgetown University |
Washington |
DC |
US |
|
|
Family ID: |
1000005384180 |
Appl. No.: |
16/757195 |
Filed: |
October 19, 2018 |
PCT Filed: |
October 19, 2018 |
PCT NO: |
PCT/US18/56608 |
371 Date: |
April 17, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62574993 |
Oct 20, 2017 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 38/1767 20130101;
A61K 31/02 20130101; A61P 11/00 20180101; A61K 9/0082 20130101 |
International
Class: |
A61K 38/17 20060101
A61K038/17; A61K 31/02 20060101 A61K031/02; A61K 9/00 20060101
A61K009/00; A61P 11/00 20060101 A61P011/00 |
Claims
1. A composition comprising silk fibroin and a perfluorocarbon.
2. The composition of claim 1, further comprising a
pharmaceutically acceptable carrier.
3. The composition of claim 1 or 2, wherein the perfluorocarbon is
selected from the group consisting of
perfluoro-n-butyltetrahydrofuran, perfluorodichlorooctane,
perfluorobischlorobutylether, perfluorodecalin,
perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
4. The composition of any one of claims 1-3, wherein the
perfluorocarbon is perfluorooctylbromide.
5. The composition of any one of claims 1-4, wherein the ratio of
silk fibroin to perfluorocarbon is about 1:4 to 1:1 (v/v).
6. The composition of any one of claims 1-5, further comprising a
drug, enzyme, antibody, or vaccine.
7. A composition comprising silk fibroin and a surfactant.
8. The composition of claim 7, further comprising a
pharmaceutically acceptable carrier.
9. The composition of claim 7 or 8, wherein the surfactant is
selected from the group consisting of egg yolk phospholipid,
polyalkyleneoxides, 1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof.
10. The composition of any one of claims 7-9, wherein the
surfactant is calfactant.
11. The composition of any one of claims 7-10, wherein the ratio of
silk fibroin to surfactant is about 1:6 to 1:2.5 (v/v).
12. The composition of any one of claims 7-11, further comprising a
drug, enzyme, antibody, or vaccine.
13. A composition comprising silk fibroin, a perfluorocarbon, and a
surfactant.
14. The composition of claim 13, further comprising a
pharmaceutically acceptable carrier.
15. The composition of claim 13 or 14, wherein the perfluorocarbon
is selected from the group consisting of
perfluoro-n-butyltetrahydrofuran, perfluorodichlorooctane,
perfluorobischlorobutylether, perfluorodecalin,
perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
16. The composition of any one of claims 13-15, wherein the
perfluorocarbon is perfluorooctylbromide.
17. The composition of any one of claims 13-16, wherein the
surfactant is selected from the group consisting of egg yolk
phospholipid, polyalkyleneoxides, 1,2-dialkylglycero-3-phosphoryl
cholines, 1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof.
18. The composition of any one of claims 13-17, wherein the
surfactant is calfactant.
19. The composition of any one of claims 13-18, wherein the ratio
of silk fibroin to perfluorocarbon is about 1:4 to 1:1 (v/v).
20. The composition of any one of claims 13-19, wherein the ratio
of silk fibroin to surfactant is about 1:6 to 1:2.5 (v/v).
21. The composition of any one of claims 13-20, further comprising
a drug, enzyme, antibody, or vaccine.
22. A method of making the composition of claim 1, comprising
placing the silk fibroin and the perfluorocarbon in an aqueous
medium.
23. A method of making the composition of claim 7, comprising
placing the silk fibroin and the surfactant in an aqueous
medium.
24. A method of making the composition of claim 13, comprising
placing the silk fibroin, the perfluorocarbon, and the surfactant
in an aqueous medium.
25. A method of treating a lung disease or lung condition,
comprising administering a therapeutically effective amount of the
composition of claim 1 to an airway of a subject in need
thereof.
26. The method of claim 25, wherein the subject is a human.
27. The method of claim 26, wherein the subject is a preterm infant
or an infant.
28. The method of any one of claims 25-27, wherein the lung disease
or lung condition is selected from the group consisting of
respiratory failure, surfactant deficiency, respiratory distress
syndrome (RDS), adult respiratory distress syndrome (ARDS),
meconium aspiration syndrome, hyaline membrane disease, pneumonia,
cystic fibrosis, idiopathic pulmonary fibrosis, atelectasis, and
any combination thereof.
29. The method of claim 28, wherein the lung disease or lung
condition is respiratory distress syndrome (RDS).
30. The method of any one of claims 25-29, wherein the
administering comprises liquid ventilation.
31. The method of claim 30, wherein the liquid ventilation is total
liquid ventilation.
32. The method of claim 30, wherein the liquid ventilation is
partial liquid ventilation.
33. A method of treating a lung disease or lung condition,
comprising administering a therapeutically effective amount of the
composition of claim 7 to an airway of a subject in need
thereof.
34. The method of claim 33, wherein the subject is a human.
35. The method of claim 34, wherein the subject is a preterm infant
or an infant.
36. The method of any one of claims 33-35, wherein the lung disease
or lung condition is selected from the group consisting of
respiratory failure, surfactant deficiency, respiratory distress
syndrome (RDS), adult respiratory distress syndrome (ARDS),
meconium aspiration syndrome, hyaline membrane disease, pneumonia,
cystic fibrosis, idiopathic pulmonary fibrosis, atelectasis, and
any combination thereof.
37. The method of claim 36, wherein the lung disease or lung
condition is respiratory distress syndrome (RDS).
38. The method of any one of claims 33-37, wherein the
administering comprises liquid ventilation.
39. The method of claim 38, wherein the liquid ventilation is total
liquid ventilation.
40. The method of claim 38, wherein the liquid ventilation is
partial liquid ventilation.
41. A method of treating a lung disease or lung condition,
comprising administering a therapeutically effective amount of the
composition of claim 13 to an airway of a subject in need
thereof.
42. The method of claim 41, wherein the subject is a human.
43. The method of claim 42, wherein the subject is a preterm infant
or an infant.
44. The method of any one of claims 41-43, wherein the lung disease
or lung condition is selected from the group consisting of
respiratory failure, surfactant deficiency, respiratory distress
syndrome (RDS), adult respiratory distress syndrome (ARDS),
meconium aspiration syndrome, hyaline membrane disease, pneumonia,
cystic fibrosis, idiopathic pulmonary fibrosis, atelectasis, and
any combination thereof.
45. The method of claim 44, wherein the lung disease or lung
condition is respiratory distress syndrome (RDS).
46. The method of any one of claims 41-45, wherein the
administering comprises liquid ventilation.
47. The method of claim 46, wherein the liquid ventilation is total
liquid ventilation.
48. The method of claim 46, wherein the liquid ventilation is
partial liquid ventilation.
49. A kit comprising silk fibroin in a first container and a
perfluorocarbon in a second container.
50. A kit comprising silk fibroin in a first container and a
surfactant in a second container.
51. A kit comprising silk fibroin in a first container, a
perfluorocarbon in a second container, and a surfactant in a third
container.
Description
RELATED APPLICATIONS
[0001] This application claims benefit of priority to U.S.
Provisional Patent Application No. 62/574,993, filed Oct. 20, 2017,
the entire content of which is incorporated herein by
reference.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted electronically in ASCII format and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Oct. 11, 2018, is named 606117_GUS-021PC_ST25.txt and is 392
bytes in size.
BACKGROUND
[0003] Liquid silk is produced by a variety of insects and spiders.
The best characterized silks are cocoon silk from the domesticated
silkworm Bombyx mori. Bombyx mori silk is composed of a core
protein, silk fibroin, and a glue-like coating consisting of a
nonfilamentous protein, sericin. The core protein of Bombyx mori
silk is made from two structural proteins, fibroin heavy chain
(.about.325 kDa) and light chain (.about.25 kDa). The fibroin heavy
chain protein consists of layers of antiparallel beta sheets, and
its primary structure mainly consists of the recurrent hydrophobic
amino acid sequence Gly-Ser-Gly-Ala-Gly-Ala (GSGAGA) (SEQ ID NO:1).
Silk fibroin is a natural or synthetic polymer used in textile
production, medical sutures, and more recently as a scaffold for
tissue regeneration and other medical applications.
[0004] Because of their characteristics, perfluorochemicals have
been proposed for use as perfusates for organs, as blood
substitutes, and as liquids for liquid lavaging or liquid
ventilation of the lungs. Perfluorocarbons (PFCs) are chemically
inert materials that are known to enable oxygen transport in
mammalian systems. For example, rats have been shown to survive
total immersion in a liquid perfluorochemical saturated with
oxygen. The high solubility of oxygen in most perfluorochemicals
enables the rat to "breathe" the perfluorochemical. This process,
and its variants, are commonly referred to as liquid ventilation.
In one form of liquid ventilation, known as perfluorochemical
assisted gas exchange (PAGE), a pure fluorochemical liquid is
instilled into the lungs of an animal in an amount equal to the
functional residual capacity of the lungs. The animal then is
connected to a mechanical ventilator which delivers tidal volumes
of a breathable gas to the lungs. However, the PAGE technique is
extremely limited with respect to the ability to deliver drugs to
the lungs. Because perfluorochemicals are both hydrophobic and
lipophobic, very few medicaments other than halocarbon anesthetics
may be delivered to the lungs using such liquid breathing
techniques.
[0005] Surfactant therapy has substantially improved the survival
of premature infants with respiratory distress syndrome (RDS),
which results from a deficiency of pulmonary surfactant. However,
exogenous surfactant is not uniformly effective in treating preterm
infants with RDS because the surfactant fails to reach atelectatic
alveoli. Perfluorocarbons have been shown to be effective in
recruiting atelectatic areas but do not replace impaired endogenous
surfactant.
SUMMARY
[0006] It has now been discovered by the present inventor that silk
fibroin, PFC, and lung surfactant can be combined to form an
aqueous emulsion, and that such preparation can (i) prolong the
effect of lung surfactant, (ii) lead to a substantially more rapid
improvement of oxygenation compared to lung surfactant alone, and
(iii) lead to a more homogenous distribution of surfactant.
Importantly, such preparation also provides substantial improvement
in lung volumes, lung compliance, oxygenation, and ventilation
compared to surfactant or PFC alone. This mixture will improve
delivery to lung of both surfactant and PFC, and it will find use
in the treatment of various lung diseases and conditions, notably
including RDS in preterm infants. This mixture may also be used as
lung lavage in other infant disease conditions such as MAS
(meconium aspiration syndrome) and/or pneumonia and to improve lung
function.
[0007] An aspect of the invention is a composition comprising silk
fibroin and a perfluorocarbon. In certain embodiments, the
perfluorocarbon is selected from the group consisting of
perfluoro-n-butyltetrahydrofuran, perfluorodichlorooctane,
perfluorobischlorobutylether, perfluorodecalin,
perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0008] An aspect of the invention is a composition comprising silk
fibroin and a surfactant. In certain embodiments, the surfactant is
selected from the group consisting of egg yolk phospholipid,
polyalkyleneoxides, 1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0009] An aspect of the invention is a composition comprising silk
fibroin, a perfluorocarbon, and a surfactant. In certain
embodiments, the perfluorocarbon is selected from the group
consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof. In
certain embodiments, the surfactant is selected from the group
consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0010] In certain embodiments, any one of the foregoing
compositions further comprises a drug, enzyme, antibody, or
vaccine.
[0011] An aspect of the invention is a method of making any one of
the foregoing compositions.
[0012] An aspect of the invention is a method of treating a lung
disease or lung condition, comprising administering a
therapeutically effective amount of any one of the aforementioned
compositions to an airway of a subject in need thereof. In certain
embodiments, the subject is a human. In certain embodiments, the
subject is a preterm infant. In certain embodiments, the subject is
an infant. In certain embodiments, the lung disease or lung
condition is selected from the group consisting of respiratory
failure, surfactant deficiency, respiratory distress syndrome
(RDS), adult respiratory distress syndrome (ARDS), meconium
aspiration syndrome, hyaline membrane disease, pneumonia, cystic
fibrosis, idiopathic pulmonary fibrosis, atelectasis, and any
combination thereof. In certain embodiments, the lung disease or
lung condition is respiratory distress syndrome (RDS). In certain
embodiments, the administering comprises liquid ventilation. In
certain embodiments, the administering comprises lung lavage, e.g.,
bronchoalveolar lavage (BAL).
[0013] An aspect of the invention is a kit comprising components of
any one of the aforementioned compositions.
BRIEF DESCRIPTION OF THE DRAWINGS
[0014] FIG. 1A is a photographic image depicting an aqueous mixture
of silk fibroin and PFC (perfluorooctylbromide).
[0015] FIG. 1B is a photographic image depicting an aqueous mixture
of silk fibroin and surfactant.
[0016] FIG. 1C is a photographic image depicting an aqueous mixture
of PFC and surfactant.
[0017] FIG. 1D is a photographic image depicting an aqueous mixture
of silk fibroin, surfactant, and PFC prior to vigorous shaking.
[0018] FIG. 1E is a photographic image depicting an aqueous mixture
of silk fibroin, surfactant, and PFC after vigorous shaking.
DETAILED DESCRIPTION
[0019] Compositions of the invention include aqueous mixtures of
silk fibroin and (a) perfluorocarbon, (b) surfactant, or (c) both
perfluorocarbon and surfactant. Each of the components silk
fibroin, perfluorocarbon, and surfactant, is deemed to be an active
agent. As disclosed herein, in certain embodiments the compositions
or mixtures of the invention can further comprise one or more
additional agents, including other active pharmaceutical
ingredients (APIs). Also as disclosed herein, the compositions of
the invention are believed to be useful in the treatment of certain
lung diseases and lung conditions, including ARDS.
[0020] The silk fibroin component is believed to facilitate the
delivery, and therefore enhance the efficacy, of each component
with which it is combined in accordance with the compositions of
the invention. In fact, it is believed that the combination of silk
fibroin, perfluorocarbon, and surfactant exhibits synergistic
biological properties over any pair of these three components.
[0021] In certain embodiments, the aqueous mixtures are
dispersions. In certain embodiments, the aqueous mixtures are
emulsions. In certain embodiments, the aqueous mixtures are
microemulsions. As will be discussed below, the various components
can be combined in various relative amounts or ratios.
[0022] As used herein, a "silk fibroin" or "fibroin" refers to
natural or recombinant silk fibroin. In certain embodiments, the
silk fibroin is derived from the domesticated Bombyx mori silkworm.
In certain embodiments, "silk fibroin" refers to an aqueous
solution containing natural or recombinant silk fibroin protein.
See, for example, U.S. Pat. No. 7,635,755, International Patent
Application Publication No. WO 97/08315, and U.S. Pat. No.
5,245,012, the entire contents of which are incorporated herein by
reference.
[0023] Silk is a well described natural fiber produced by the
silkworm, Bombyx mori, which has been used traditionally in the
form of threads in textiles for thousands of years. This silk
contains a fibrous protein termed fibroin (both heavy and light
chains) that form the thread core, and glue-like proteins termed
sericin that surround the fibroin fibers to cement them together.
The fibroin is a highly insoluble protein containing up to 90% the
amino acids glycine, alanine and serine leading to .beta.-pleated
sheet formation in the fibers (Asakura, et al., Encyclopedia of
Agricultural Science, Arntzen, C. J., Ritter, E. M. Eds.; Academic
Press: New York, N.Y., 1994; Vol. 4, pp 1-11).
[0024] Silk fibroin has been proposed as a vehicle for drug
delivery. See, for example, Wenk et al. Biomaterials 31: 1403-13
(2010), and Wenk et al. J Control Release 150: 128-41 (2011), the
entire contents of which are incorporated herein by reference. Silk
fibroin has unique self-assembly process (driving force:
hydrophobicity and electrostatic interactions), and it acts like a
cocoon for biological matter. Cells show normal growth and
proliferation in the presence of silk fibroin. A common and
straightforward way to incorporate drugs into the fabrication of
silk fibroin delivery system is by dissolving or mixing them
directly into silk fibroin solution. Addition of enzymes or
antibodies or vaccines to liquid silk solution (the self-assembly
process) preserves the biological function of these compounds.
[0025] In certain embodiments, the silk fibroin is provided as a
commercial product, for example, silk fibroin 50 mg/mL in aqueous
solution (Sigma-Aldrich).
[0026] As used herein, a "perfluorocarbon" or "PFC" refers to a
hydrocarbon, or optionally a substituted hydrocarbon, in which all
C--H bonds have been replaced by C--F bonds. Perfluorocarbons
include perfluoroalkanes, perfluoroalkenes, perfluoroalkynes, and
perfluoroaromatic compounds. In certain embodiments, a
"perfluorocarbon" refers to a perfluoroalkane. The alkane portion
can be unbranched, branched, or cyclical. There are five
perfluoroalkane gases: tetrafluoromethane (bp -128.degree. C.),
hexafluoroethane (bp -78.2.degree. C.), octafluoropropane (bp
-36.5.degree. C.), perfluoro-n-butane (bp -2.2.degree. C.), and
perfluoro-iso-butane (bp -1.degree. C.). Nearly all other
fluoroalkanes are liquids.
[0027] PFC liquids have low surface tension and at atmospheric
pressure, large amounts of oxygen and carbon dioxide dissolve in
them. For example, PFC carries 20 times more oxygen than
saline/water and twice that of blood. The 02 diffusion rate from
PFC-filled alveoli to the alveolar capillaries is quite high.
Clear, colorless, odorless, non-conducting, and non-flammable, PFCs
are insoluble in water and only sparingly soluble in lipid. PFCs
are not metabolized in body tissues.
[0028] In certain embodiments, a perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof. In
certain embodiments, a perfluorocarbon is
perfluorooctylbromide.
[0029] A perfluorocarbon may be presented in a gas or liquid form.
Liquid forms may include a water-in-perfluorocarbon stable liquid
dispersion or a perfluorocarbon-in-water microemulsion. Homogenous
water-in-perfluorochemical stable liquid dispersions are disclosed
in U.S. Pat. No. 5,770,585, the entire content of which is
incorporated herein by reference.
[0030] In certain embodiments, a perfluorocarbon is provided as a
commercial product, for example, LiquiVent.RTM.
(perfluorooctylbromide; perflubron; Origen Biomedical, Austin,
Tex.).
[0031] As used herein, a "surfactant" refers to a compound that
lowers the surface tension between two liquids or between a liquid
and a solid. In certain embodiments, a surfactant is a pulmonary
surfactant.
[0032] Pulmonary surfactant is a surface-active phospholipoprotein
complex formed by type II alveolar cells of the lung. The proteins
and lipids that make up the surfactant have both hydrophilic and
hydrophobic regions. By adsorbing to the air-water interface of
alveoli, with hydrophilic head groups in the water and the
hydrophobic tails facing towards the air, the main lipid component
of surfactant, dipalmitoylphosphatidylcholine (DPPC), reduces
surface tension. Pulmonary surfactant increases pulmonary
compliance and facilitates recruitment of collapsed (atelectatic)
airways. Most preterm infants are surfactant-deficient at birth and
as a result have respiratory problems, including respiratory
distress syndrome (RDS). RDS is a major cause of premature infant
morbidity and mortality.
[0033] Pulmonary surfactants generally include synthetic pulmonary
surfactants and animal-derived pulmonary surfactants. Both
synthetic pulmonary surfactants and animal-derived pulmonary
surfactants are useful in the invention.
[0034] Synthetic pulmonary surfactants include, without limitation,
colfosceril palmitate (Exosurf), a mixture of
dipalmitoylphosphatidylcholine (DPPC) with hexadecanol and
tyloxapol; pumactant, a mixture of DPPC and phosphatidylglycerol
(PG); KL-4, a mixture of DPPC, palmitoyl-oleoyl
phosphatidylglycerol, and palmitic acid, combined with a 21 amino
acid synthetic peptide that mimics the structural characteristics
of surfactant protein B (SP-B); venticute, a mixture of DPPC, PG,
palmitic acid, and recombinant surfactant protein C (SP-C); and
lucinactant (Surfaxin), a mixture of KL4 acetate,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (as the sodium
salt), and palmitic acid. PG consists of an L-glycerol 3-phosphate
backbone ester-bonded to either saturated or unsaturated fatty
acids on carbons 1 and 2.
[0035] Animal-derived pulmonary surfactants include, without
limitation, beractant, Alveofact (extracted from cow lung lavage
fluid, manufactured by Boehringer Ingelheim), Survanta (extracted
from minced cow lung with additional DPPC, palmitic acid and
tripalmitin, manufactured by Abbvie), Beraksurf (extracted from
minced cow lung with additional DPPC, palmitic acid and tripalmitin
manufactured by Tekzima), calfactant (Infasurf, extracted from calf
lung lavage), and proactant alfa (Curosurf.RTM., extracted from
material derived from minced pig lung).
[0036] In certain embodiments, the surfactant is provided as a
commercial product, for example, calfactant (Infasurf.RTM.).
[0037] As used herein, a "lung disease" refers to any congenital or
acquired lung disease characterized by impairment of gas exchange.
Lung diseases include, without limitation, surfactant deficiency,
respiratory distress syndrome (RDS), adult respiratory distress
syndrome (ARDS), meconium aspiration syndrome, pneumonia, cystic
fibrosis, idiopathic pulmonary fibrosis, bronchitis, emphysema, and
lung cancer.
[0038] As used herein, a "lung condition" refers to any congenital
or acquired lung condition, other than a lung disease,
characterized by impairment of gas exchange. Lung conditions
include, without limitation, atelectasis.
[0039] As used herein, a "preterm infant" refers to an infant born
at less than 32 weeks gestational age.
[0040] As used herein, an "infant" refers to a human baby less than
one year old. In certain embodiments, an infant is a neonate or
newborn.
[0041] As used herein, "respiratory failure" refers to compromise
of lung gas exchange from any cause, characterized by diminished
PaO.sub.2 and/or increased PaCO.sub.2 (without treatment) compared
to normal values. In certain embodiments, respiratory failure is
characterized by PaO.sub.2 (without treatment) of less than or
equal to 50 mm Hg. In certain embodiments, respiratory failure is
sufficiently severe as to require supplemental oxygen. In certain
embodiments, respiratory failure is sufficiently severe as to
require mechanical ventilation. Respiratory failure can be acute or
chronic.
[0042] As used herein, "surfactant deficiency" refers to any
congenital or acquired deficiency of pulmonary surfactant. In
certain embodiments, surfactant deficiency is accompanied by
respiratory failure. Surfactant deficiency can be natural or it can
be experimentally induced.
[0043] As used herein, the terms "respiratory distress syndrome" or
"RDS" refer to a breathing disorder that affects newborns,
especially preterm infants. It is believed to be caused by
pulmonary surfactant deficiency due at least in part to immaturity
of the type II alveolar cells which are the principal source of
pulmonary surfactant. This syndrome is also variously known as
infant respiratory distress syndrome (IRDS), neonatal respiratory
distress syndrome, surfactant deficiency disorder, and hyaline
membrane disease.
[0044] Acute respiratory distress syndrome (ARDS) is
life-threatening condition that can develop principally in adults
and is characterized by inflammation of and abnormal accumulation
of fluid in alveoli of the lung. ARDS may be seen in the setting of
a critical illness, e.g., severe pulmonary (pneumonia) or systemic
infection (sepsis), following trauma, multiple blood transfusions,
severe burns, severe pancreatitis, near-drowning, drug reactions,
or inhalation injuries. ARDS is characterized by: lung injury of
acute onset, within 1 week of an apparent clinical insult and with
progression of respiratory symptoms; bilateral opacities on chest
imaging (chest X-ray or CT) not explained by other lung pathology;
respiratory failure not explained by heart failure or volume
overload; and decreased PaO.sub.2/FiO.sub.2 ratio.
[0045] As used herein, "liquid ventilation" refers to a technique
of mechanical ventilation in which the lungs are insufflated with
an oxygenated perfluorochemical liquid rather than an
oxygen-containing gas mixture. For a review, see Tawfic et al.,
Oman Med J 26:(1):4-9 (2011).
[0046] As used herein, "total liquid ventilation" refers to a type
of liquid ventilation in which the lungs are filled with PFC to a
volume equivalent to the functional residual capacity,
approximately 30 mL/kg, and a liquid ventilator is used to generate
tidal breathing with perfluorocarbon.
[0047] As used herein, "partial liquid ventilation" refers to a
type of liquid ventilation in which tracheal instillation of PFC
liquid is used in combination with conventional gas mechanical
ventilation.
Compositions
[0048] An aspect of the invention is a composition comprising silk
fibroin and a perfluorocarbon. In certain embodiments, the silk
fibroin is provided as an aqueous solution. In certain embodiments,
the perfluorocarbon is provided as an aqueous solution. In certain
embodiments, the composition is an aqueous solution comprising silk
fibroin and a perfluorocarbon. In certain embodiments, the
composition is an aqueous dispersion comprising silk fibroin and a
perfluorocarbon. In certain embodiments, the composition is an
aqueous emulsion comprising silk fibroin and a perfluorocarbon.
[0049] In certain embodiments, the composition further comprises a
pharmaceutically acceptable carrier, such as is described elsewhere
herein.
[0050] In certain embodiments, the perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0051] In certain embodiments, the perfluorocarbon is
perfluorooctylbromide (1-Bromoperfluorooctane), e.g.,
perflubron.
[0052] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:1 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to perfluorocarbon is about
1:4 (v/v). In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 (v/v). In certain embodiments, the
ratio of silk fibroin to perfluorocarbon is about 1:2 (v/v). In
certain embodiments, the ratio of silk fibroin to perfluorocarbon
is about 1:1 (v/v).
[0053] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:2 (v/v).
[0054] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:3 (v/v).
[0055] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:1 (v/v).
[0056] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:2 (v/v).
[0057] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:2 to 1:1 (v/v).
[0058] In certain embodiments, the composition further comprises
another active pharmaceutical agent (API). In certain embodiments,
said other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, a diuretic, or
deoxyribonuclease (DNAse) enzyme.
[0059] Exemplary steroids include, without limitation,
methylprednisone, beclomethasone, dexamethasone, budesonide,
flunisolide, fluticasone, and hydrocortisone.
[0060] Exemplary diuretics include, without limitation, furosemide,
bumetanide, and amiloride.
[0061] A further aspect of the invention is a composition
comprising silk fibroin and a surfactant. In certain embodiments,
the silk fibroin is provided as an aqueous solution. In certain
embodiments, the surfactant is provided as an aqueous solution. In
certain embodiments, the composition is an aqueous solution
comprising silk fibroin and a surfactant. In certain embodiments,
the composition is an aqueous dispersion comprising silk fibroin
and a surfactant. In certain embodiments, the composition is an
aqueous emulsion comprising silk fibroin and a surfactant.
[0062] In certain embodiments, the composition further comprises a
pharmaceutically acceptable carrier, such as is described elsewhere
herein.
[0063] In certain embodiments, the surfactant is a pulmonary
surfactant.
[0064] In certain embodiments, the surfactant is selected from the
group consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon. In certain
embodiments, the surfactant is calfactant.
[0065] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:2 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to surfactant is about 1:6
to 1:2.5 (v/v). In certain embodiments, the ratio of silk fibroin
to surfactant is about 1:6 (v/v). In certain embodiments, the ratio
of silk fibroin to surfactant is about 1:5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:4
(v/v). In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 (v/v). In certain embodiments, the ratio of
silk fibroin to surfactant is about 1:2.5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:2
(v/v).
[0066] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:3 (v/v).
[0067] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:4 (v/v).
[0068] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:5 (v/v).
[0069] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2 (v/v).
[0070] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2.5 (v/v).
[0071] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:3 (v/v).
[0072] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:4 (v/v).
[0073] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2 (v/v).
[0074] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2.5 (v/v).
[0075] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:3 (v/v).
[0076] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2 (v/v).
[0077] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2.5 (v/v).
[0078] In certain embodiments, the composition further comprises
another active pharmaceutical agent (API). In certain embodiments,
said other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, diuretic, or
deoxyribonuclease (DNAse) enzyme.
[0079] A yet further aspect of the invention is a composition
comprising silk fibroin, a perfluorocarbon, and a surfactant. In
certain embodiments, the silk fibroin is provided as an aqueous
solution. In certain embodiments, the perfluorocarbon is provided
as an aqueous solution. In certain embodiments, the surfactant is
provided as an aqueous solution. In certain embodiments, the
composition is an aqueous solution comprising silk fibroin, a
perfluorocarbon, and a surfactant. In certain embodiments, the
composition is an aqueous dispersion comprising silk fibroin, a
perfluorocarbon, and a surfactant. In certain embodiments, the
composition is an aqueous emulsion comprising silk fibroin, a
perfluorocarbon, and a surfactant.
[0080] In certain embodiments, the composition further comprises a
pharmaceutically acceptable carrier, such as is described elsewhere
herein.
[0081] In certain embodiments, the perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0082] In certain embodiments, the perfluorocarbon is
perfluorooctylbromide (1-Bromoperfluorooctane), e.g.,
perflubron.
[0083] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:1 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to perfluorocarbon is about
1:4 (v/v). In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 (v/v). In certain embodiments, the
ratio of silk fibroin to perfluorocarbon is about 1:2 (v/v). In
certain embodiments, the ratio of silk fibroin to perfluorocarbon
is about 1:1 (v/v).
[0084] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:2 (v/v).
[0085] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:3 (v/v).
[0086] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:1 (v/v).
[0087] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:2 (v/v).
[0088] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:2 to 1:1 (v/v).
[0089] In certain embodiments, the surfactant is a pulmonary
surfactant.
[0090] In certain embodiments, the surfactant is selected from the
group consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0091] In certain embodiments, the surfactant is calfactant.
[0092] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:2 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to surfactant is about 1:6
to 1:2.5 (v/v). In certain embodiments, the ratio of silk fibroin
to surfactant is about 1:6 (v/v). In certain embodiments, the ratio
of silk fibroin to surfactant is about 1:5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:4
(v/v). In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 (v/v). In certain embodiments, the ratio of
silk fibroin to surfactant is about 1:2.5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:2
(v/v).
[0093] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:3 (v/v).
[0094] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:4 (v/v).
[0095] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:5 (v/v).
[0096] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2 (v/v).
[0097] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2.5 (v/v).
[0098] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:3 (v/v).
[0099] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:4 (v/v).
[0100] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2 (v/v).
[0101] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2.5 (v/v).
[0102] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:3 (v/v).
[0103] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2 (v/v).
[0104] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2.5 (v/v).
[0105] In certain embodiments, the composition further comprises
another active pharmaceutical agent (API). In certain embodiments,
said other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, diuretic, or
deoxyribonuclease (DNAse) enzyme.
Methods of Making
[0106] An aspect of the invention is a method of making a
composition comprising silk fibroin and a perfluorocarbon. The
method includes the step of placing the silk fibroin and the
perfluorocarbon in an aqueous medium. In certain embodiments, the
method further includes the step of mixing the silk fibroin and the
perfluorocarbon in the aqueous medium. The mixing can be
accomplished by any suitable means, including, without limitation,
stirring, rocking, shaking, and vortexing.
[0107] In certain embodiments, the silk fibroin is provided as an
aqueous solution. In certain embodiments, the perfluorocarbon is
provided as an aqueous solution. In certain embodiments, the
composition is an aqueous solution comprising silk fibroin and a
perfluorocarbon. In certain embodiments, the composition is an
aqueous dispersion comprising silk fibroin and a perfluorocarbon.
In certain embodiments, the composition is an aqueous emulsion
comprising silk fibroin and a perfluorocarbon.
[0108] In certain embodiments, the method comprises placing the
silk fibroin and the perfluorocarbon in a pharmaceutically
acceptable carrier, such as is described elsewhere herein.
[0109] In certain embodiments, the perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0110] In certain embodiments, the perfluorocarbon is
perfluorooctylbromide, e.g., perflubron.
[0111] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:1 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to perfluorocarbon is about
1:4 (v/v). In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 (v/v). In certain embodiments, the
ratio of silk fibroin to perfluorocarbon is about 1:2 (v/v). In
certain embodiments, the ratio of silk fibroin to perfluorocarbon
is about 1:1 (v/v).
[0112] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:2 (v/v).
[0113] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:3 (v/v).
[0114] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:1 (v/v).
[0115] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:2 (v/v).
[0116] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:2 to 1:1 (v/v).
[0117] In certain embodiments, the method further comprises
contacting the silk fibroin and the perfluorocarbon with another
active pharmaceutical agent (API). In certain embodiments, said
other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, diuretic, or
deoxyribonuclease (DNAse) enzyme.
[0118] An aspect of the invention is a method of making a
composition comprising silk fibroin and a surfactant. The method
includes the step of placing the silk fibroin and the surfactant in
an aqueous medium. In certain embodiments, the method further
includes the step of mixing the silk fibroin and the surfactant in
the aqueous medium. The mixing can be accomplished by any suitable
means, including, without limitation, stirring, rocking, shaking,
and vortexing.
[0119] In certain embodiments, the silk fibroin is provided as an
aqueous solution. In certain embodiments, the surfactant is
provided as an aqueous solution. In certain embodiments, the
composition is an aqueous solution comprising silk fibroin and a
surfactant. In certain embodiments, the composition is an aqueous
dispersion comprising silk fibroin and a surfactant. In certain
embodiments, the composition is an aqueous emulsion comprising silk
fibroin and a surfactant.
[0120] In certain embodiments, the method comprises placing the
silk fibroin and the surfactant in a pharmaceutically acceptable
carrier, such as is described elsewhere herein.
[0121] In certain embodiments, the surfactant is a pulmonary
surfactant.
[0122] In certain embodiments, the surfactant is selected from the
group consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0123] In certain embodiments, the surfactant is calfactant.
[0124] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:2 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to surfactant is about 1:6
to 1:2.5 (v/v). In certain embodiments, the ratio of silk fibroin
to surfactant is about 1:6 (v/v). In certain embodiments, the ratio
of silk fibroin to surfactant is about 1:5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:4
(v/v). In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 (v/v). In certain embodiments, the ratio of
silk fibroin to surfactant is about 1:2.5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:2
(v/v).
[0125] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:3 (v/v).
[0126] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:4 (v/v).
[0127] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:5 (v/v).
[0128] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2 (v/v).
[0129] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2.5 (v/v).
[0130] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:3 (v/v).
[0131] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:4 (v/v).
[0132] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2 (v/v).
[0133] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2.5 (v/v).
[0134] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:3 (v/v).
[0135] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2 (v/v).
[0136] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2.5 (v/v).
[0137] In certain embodiments, the method further comprises
contacting the silk fibroin and the surfactant with another active
pharmaceutical agent (API). In certain embodiments, said other API
is a drug, enzyme, antibody, or vaccine. In certain embodiments,
said other API is a steroid, diuretic, or deoxyribonuclease (DNAse)
enzyme.
[0138] A yet further aspect of the invention is a method of making
a composition comprising silk fibroin, a perfluorocarbon, and a
surfactant. The method includes the step of placing the silk
fibroin, the perfluorocarbon, and the surfactant in an aqueous
medium. In certain embodiments, the method further includes the
step of mixing the silk fibroin, the perfluorocarbon, and the
surfactant in the aqueous medium. The mixing can be accomplished by
any suitable means, including, without limitation, stirring,
rocking, shaking, and vortexing.
[0139] In certain embodiments, the silk fibroin is provided as an
aqueous solution. In certain embodiments, the perfluorocarbon is
provided as an aqueous solution. In certain embodiments, the
surfactant is provided as an aqueous solution. In certain
embodiments, the composition is an aqueous solution comprising silk
fibroin, a perfluorocarbon, and a surfactant. In certain
embodiments, the composition is an aqueous dispersion comprising
silk fibroin, a perfluorocarbon, and a surfactant. In certain
embodiments, the composition is an aqueous emulsion comprising silk
fibroin, a perfluorocarbon, and a surfactant.
[0140] In certain embodiments, the method comprises placing the
silk fibroin, perfluorocarbon, and surfactant in a pharmaceutically
acceptable carrier, such as is described elsewhere herein.
[0141] In certain embodiments, the perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0142] In certain embodiments, the perfluorocarbon is
perfluorooctylbromide (1-Bromoperfluorooctane), e.g.,
perflubron.
[0143] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:1 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to perfluorocarbon is about
1:4 (v/v). In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 (v/v). In certain embodiments, the
ratio of silk fibroin to perfluorocarbon is about 1:2 (v/v). In
certain embodiments, the ratio of silk fibroin to perfluorocarbon
is about 1:1 (v/v).
[0144] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:2 (v/v).
[0145] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:3 (v/v).
[0146] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:1 (v/v).
[0147] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:2 (v/v).
[0148] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:2 to 1:1 (v/v).
[0149] In certain embodiments, the surfactant is a pulmonary
surfactant.
[0150] In certain embodiments, the surfactant is selected from the
group consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0151] In certain embodiments, the surfactant is calfactant.
[0152] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:2 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to surfactant is about 1:6
to 1:2.5 (v/v). In certain embodiments, the ratio of silk fibroin
to surfactant is about 1:6 (v/v). In certain embodiments, the ratio
of silk fibroin to surfactant is about 1:5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:4
(v/v). In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 (v/v). In certain embodiments, the ratio of
silk fibroin to surfactant is about 1:2.5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:2
(v/v).
[0153] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:3 (v/v).
[0154] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:4 (v/v).
[0155] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:5 (v/v).
[0156] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2 (v/v).
[0157] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2.5 (v/v).
[0158] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:3 (v/v).
[0159] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:4 (v/v).
[0160] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2 (v/v).
[0161] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2.5 (v/v).
[0162] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:3 (v/v).
[0163] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2 (v/v).
[0164] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2.5 (v/v).
[0165] In certain embodiments, the method further comprises the
step of contacting the silk fibroin, perfluorocarbon, and
surfactant with an active pharmaceutical agent (API). In certain
embodiments, said other API is a drug, enzyme, antibody, or
vaccine. In certain embodiments, said other API is a steroid,
diuretic, or deoxyribonuclease (DNAse) enzyme.
Methods of Using
[0166] An aspect of the invention is a method of treating a lung
disease or lung condition, comprising administering a
therapeutically effective amount of a composition comprising silk
fibroin and a perfluorocarbon to an airway of a subject in need
thereof.
[0167] As used herein, the terms "treat" and "treating" refer to
reducing or ameliorating at least one sign or symptom of a disease
or condition of a subject having said disease or condition. In
certain embodiments, the terms "treat" and "treating" encompass
curing a disease or condition of a subject having said disease or
condition.
[0168] As used herein, the term "therapeutically effective amount"
refers to an amount sufficient to achieve a desired therapeutic
result in a subject. For example, a therapeutically effective
amount can refers to an amount sufficient to reduce or ameliorate
at least one sign or symptom of a disease or condition of a subject
having said disease or condition.
[0169] As used herein, an "airway" refers to any conducting or
gas-exchanging anatomical structure. In certain embodiments, an
airway is any one or combination of trachea, bronchi, bronchioles,
and alveoli.
[0170] As used herein, a "subject" refers to a living animal. In
certain embodiments, a subject is a mammal. In certain embodiments,
a subject is a mammal selected from the group consisting of mice,
rats, hamsters, guinea pigs, rabbits, goats, sheep, cats, dogs,
pigs, horses, cows, and non-human primates. In certain embodiments,
a subject is a human. In some embodiments, a subject is a preterm
infant. In some embodiments, a subject is an infant. In some
embodiments, a subject is a human 1 to 18 years of age. In some
embodiments, a subject is an adult human.
[0171] In certain embodiments, the silk fibroin is provided as an
aqueous solution. In certain embodiments, the perfluorocarbon is
provided as an aqueous solution. In certain embodiments, the
composition is an aqueous solution comprising silk fibroin and a
perfluorocarbon. In certain embodiments, the composition is an
aqueous dispersion comprising silk fibroin and a perfluorocarbon.
In certain embodiments, the composition is an aqueous emulsion
comprising silk fibroin and a perfluorocarbon.
[0172] In certain embodiments, the composition further comprises a
pharmaceutically acceptable carrier, such as is described elsewhere
herein.
[0173] In certain embodiments, the perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0174] In certain embodiments, the perfluorocarbon is
perfluorooctylbromide (1-Bromoperfluorooctane), e.g.,
perflubron.
[0175] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:1 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to perfluorocarbon is about
1:4 (v/v). In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 (v/v). In certain embodiments, the
ratio of silk fibroin to perfluorocarbon is about 1:2 (v/v). In
certain embodiments, the ratio of silk fibroin to perfluorocarbon
is about 1:1 (v/v).
[0176] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:2 (v/v).
[0177] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:3 (v/v).
[0178] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:1 (v/v).
[0179] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:2 (v/v).
[0180] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:2 to 1:1 (v/v).
[0181] In certain embodiments, the composition further comprises
another active pharmaceutical agent (API). In certain embodiments,
said other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, diuretic, or
deoxyribonuclease (DNAse) enzyme.
[0182] In certain embodiments, the subject is a preterm infant.
[0183] In certain embodiments, the subject is an infant.
[0184] In certain embodiments, the subject is an adult human.
[0185] In certain embodiments, the lung disease or lung condition
is selected from the group consisting of respiratory failure,
surfactant deficiency, respiratory distress syndrome (RDS), adult
respiratory distress syndrome (ARDS), meconium aspiration syndrome,
hyaline membrane disease, pneumonia, cystic fibrosis, idiopathic
pulmonary fibrosis, atelectasis, and any combination thereof.
[0186] In certain embodiments, the lung disease or lung condition
is respiratory distress syndrome (RDS).
[0187] In certain embodiments, the administering comprises liquid
ventilation.
[0188] In certain embodiments, the liquid ventilation is total
liquid ventilation.
[0189] In certain embodiments, the liquid ventilation is partial
liquid ventilation.
[0190] A further aspect of the invention is a method of treating a
lung disease or lung condition, comprising administering a
therapeutically effective amount of a composition comprising silk
fibroin and a surfactant to an airway of a subject in need
thereof.
[0191] In certain embodiments, the silk fibroin is provided as an
aqueous solution. In certain embodiments, the surfactant is
provided as an aqueous solution. In certain embodiments, the
composition is an aqueous solution comprising silk fibroin and a
surfactant. In certain embodiments, the composition is an aqueous
dispersion comprising silk fibroin and a surfactant. In certain
embodiments, the composition is an aqueous emulsion comprising silk
fibroin and a surfactant.
[0192] In certain embodiments, the composition further comprises a
pharmaceutically acceptable carrier, such as is described elsewhere
herein.
[0193] In certain embodiments, the surfactant is a pulmonary
surfactant.
[0194] In certain embodiments, the surfactant is selected from the
group consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0195] In certain embodiments, the surfactant is calfactant.
[0196] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:2 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to surfactant is about 1:6
to 1:2.5 (v/v). In certain embodiments, the ratio of silk fibroin
to surfactant is about 1:6 (v/v). In certain embodiments, the ratio
of silk fibroin to surfactant is about 1:5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:4
(v/v). In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 (v/v). In certain embodiments, the ratio of
silk fibroin to surfactant is about 1:2.5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:2
(v/v).
[0197] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:3 (v/v).
[0198] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:4 (v/v).
[0199] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:5 (v/v).
[0200] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2 (v/v).
[0201] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2.5 (v/v).
[0202] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:3 (v/v).
[0203] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:4 (v/v).
[0204] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2 (v/v).
[0205] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2.5 (v/v).
[0206] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:3 (v/v).
[0207] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2 (v/v).
[0208] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2.5 (v/v).
[0209] In certain embodiments, the composition further comprises
another active pharmaceutical agent (API). In certain embodiments,
said other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, diuretic, or
deoxyribonuclease (DNAse) enzyme.
[0210] In certain embodiments, the subject is a preterm infant.
[0211] In certain embodiments, the subject is an infant. In certain
embodiments, the subject is an adult human.
[0212] In certain embodiments, the lung disease or lung condition
is selected from the group consisting of respiratory failure,
surfactant deficiency, respiratory distress syndrome (RDS), adult
respiratory distress syndrome (ARDS), meconium aspiration syndrome,
hyaline membrane disease, pneumonia, cystic fibrosis, idiopathic
pulmonary fibrosis, atelectasis, and any combination thereof.
[0213] In certain embodiments, the lung disease or lung condition
is respiratory distress syndrome (RDS).
[0214] In certain embodiments, the administering comprises liquid
ventilation.
[0215] In certain embodiments, the liquid ventilation is total
liquid ventilation.
[0216] In certain embodiments, the liquid ventilation is partial
liquid ventilation.
[0217] A yet further aspect of the invention is a method of
treating a lung disease or lung condition, comprising administering
a therapeutically effective amount of a composition comprising silk
fibroin, a perfluorocarbon, and a surfactant to an airway of a
subject in need thereof.
[0218] In certain embodiments, the silk fibroin is provided as an
aqueous solution. In certain embodiments, the perfluorocarbon is
provided as an aqueous solution. In certain embodiments, the
surfactant is provided as an aqueous solution. In certain
embodiments, the composition is an aqueous solution comprising silk
fibroin, a perfluorocarbon, and a surfactant. In certain
embodiments, the composition is an aqueous dispersion comprising
silk fibroin, a perfluorocarbon, and a surfactant. In certain
embodiments, the composition is an aqueous emulsion comprising silk
fibroin, a perfluorocarbon, and a surfactant.
[0219] In certain embodiments, the composition further comprises a
pharmaceutically acceptable carrier, such as is described elsewhere
herein.
[0220] In certain embodiments, the perfluorocarbon is selected from
the group consisting of perfluoro-n-butyltetrahydrofuran,
perfluorodichlorooctane, perfluorobischlorobutylether,
perfluorodecalin, perfluoromethyldecalin, perfluorodimethyldecalin,
perfluorodimethyladamantane, perfluorooctylbromide,
perfluoro-4-methyl-octahydroquinolidizine,
perfluoro-N-methyl-decahydroquinoline, F-methyl-1-oxa-decalin,
perfluoro-bicyclo(5.3.0)-decane, perfluorooctahydroquinolidizine,
perfluoro-5,6-dihydro-5-decene, and perfluoro-4,5-dihydro-4-octene,
chlorinated perfluorocarbons, and any combination thereof.
[0221] In certain embodiments, the perfluorocarbon is
perfluorooctylbromide (1-Bromoperfluorooctane), e.g.,
perflubron.
[0222] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:1 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to perfluorocarbon is about
1:4 (v/v). In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 (v/v). In certain embodiments, the
ratio of silk fibroin to perfluorocarbon is about 1:2 (v/v). In
certain embodiments, the ratio of silk fibroin to perfluorocarbon
is about 1:1 (v/v).
[0223] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:2 (v/v).
[0224] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:4 to 1:3 (v/v).
[0225] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:1 (v/v).
[0226] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:3 to 1:2 (v/v).
[0227] In certain embodiments, the ratio of silk fibroin to
perfluorocarbon is about 1:2 to 1:1 (v/v).
[0228] In certain embodiments, the surfactant is a pulmonary
surfactant.
[0229] In certain embodiments, the surfactant is selected from the
group consisting of egg yolk phospholipid, polyalkyleneoxides,
1,2-dialkylglycero-3-phosphoryl cholines,
1,3-dialkylglycero-2-phosphoryl cholines, perfluorinated
polyoxyethylenes, and any combination thereof. In certain
embodiments, the surfactant is not a perfluorocarbon.
[0230] In certain embodiments, the surfactant is calfactant.
[0231] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:2 (v/v). For example, in certain
embodiments, the ratio of silk fibroin to surfactant is about 1:6
to 1:2.5 (v/v). In certain embodiments, the ratio of silk fibroin
to surfactant is about 1:6 (v/v). In certain embodiments, the ratio
of silk fibroin to surfactant is about 1:5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:4
(v/v). In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 (v/v). In certain embodiments, the ratio of
silk fibroin to surfactant is about 1:2.5 (v/v). In certain
embodiments, the ratio of silk fibroin to surfactant is about 1:2
(v/v).
[0232] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:3 (v/v).
[0233] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:4 (v/v).
[0234] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:6 to 1:5 (v/v).
[0235] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2 (v/v).
[0236] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:2.5 (v/v).
[0237] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:3 (v/v).
[0238] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:5 to 1:4 (v/v).
[0239] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2 (v/v).
[0240] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:2.5 (v/v).
[0241] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:4 to 1:3 (v/v).
[0242] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2 (v/v).
[0243] In certain embodiments, the ratio of silk fibroin to
surfactant is about 1:3 to 1:2.5 (v/v).
[0244] In certain embodiments, the composition further comprises
another active pharmaceutical agent (API). In certain embodiments,
said other API is a drug, enzyme, antibody, or vaccine. In certain
embodiments, said other API is a steroid, diuretic, or
deoxyribonuclease (DNAse) enzyme.
[0245] In certain embodiments, the subject is a human.
[0246] In certain embodiments, the subject is a preterm infant.
[0247] In certain embodiments, the subject is an infant.
[0248] In certain embodiments, the subject is an adult human.
[0249] In certain embodiments, the lung disease or lung condition
is selected from the group consisting of respiratory failure,
surfactant deficiency, respiratory distress syndrome (RDS), adult
respiratory distress syndrome (ARDS), meconium aspiration syndrome,
hyaline membrane disease, pneumonia, cystic fibrosis, idiopathic
pulmonary fibrosis, atelectasis, and any combination thereof.
[0250] In certain embodiments, the lung disease or lung condition
is respiratory distress syndrome (RDS).
[0251] In certain embodiments, the administering comprises liquid
ventilation.
[0252] In certain embodiments, the liquid ventilation is total
liquid ventilation.
[0253] In certain embodiments, the liquid ventilation is partial
liquid ventilation.
Formulation and Dosing
[0254] Compositions of the invention can be formulated for
administration to lung. For example, a composition of the invention
can be formulated with a pharmaceutically acceptable carrier,
wherein the composition is provided in a therapeutically effective
amount within a volume suitable for administration to an airway of
a subject to be treated.
[0255] As used herein, a "pharmaceutically acceptable carrier"
refers to a biologically compatible aqueous fluid, e.g., water,
normal saline, Ringer's solution, solution buffered to physiologic
pH (pH 6.8-7.4).
[0256] The pharmaceutically acceptable carrier can optionally
include any one or more of salts, buffering agents, osmotically
active agents, preservatives, and coloring agents, such as are well
known in the pharmaceutical arts. See, e.g., Remington's The
Science and Practice of Pharmacy, Lloyd V. Allen, Jr, editor,
Philadelphia, Pa.: Pharmaceutical Press. 2012.
[0257] For use in the lung, a suitable amount (volume) of
composition of the invention so formulated can be instilled into an
airway of the lung, e.g., via an endotracheal tube. Dosing may
involve any one or combination of (i) lavage, (ii) partial liquid
ventilation, and (iii) total liquid ventilation.
[0258] Administration can be accomplished in a single dose or in
multiple (i.e., two or more) doses. When the administration is
accomplished in multiple doses, each dose can be the same, or at
least one dose can differ from another dose.
[0259] In certain embodiments, dosing can be based on corresponding
surfactant alone.
[0260] In certain embodiments, dosing can be based on corresponding
PFC alone.
[0261] In certain embodiments, the composition is administered once
daily, twice daily, three times daily, four times daily, or more.
In certain embodiments, the composition is administered every other
day, every third day, every fourth day, every fifth day, every
sixth day, or every seventh day.
[0262] Generally, the administering physician or medical
professional will be able to assess and adjust the dose and
frequency of dosing based on such parameters as body weight, lung
volumes, age, general medical condition, other medical conditions,
and laboratory values such as O.sub.2 saturation, arterial blood
gases (ABGs), and radiographic examination of the lungs.
Kits
[0263] An aspect of the invention is a kit comprising silk fibroin
in a first container and a perfluorocarbon in a second container.
The kit may be conveniently presented with the components packaged
together, for example in a box, tray, or foil pouch. The component
silk fibroin and perfluorocarbon can be combined by an end user to
make a composition in accordance with the invention.
[0264] An aspect of the invention is a kit comprising silk fibroin
in a first container and a surfactant in a second container. The
kit may be conveniently presented with the components packaged
together, for example in a box, tray, or foil pouch. The component
silk fibroin and surfactant can be combined by an end user to make
a composition in accordance with the invention.
[0265] An aspect of the invention is a kit comprising silk fibroin
in a first container, a perfluorocarbon in a second container, and
a surfactant in a third container. The kit may be conveniently
presented with the components packaged together, for example in a
box, tray, or foil pouch. The component silk fibroin,
perfluorocarbon, and surfactant can be combined by an end user to
make a composition in accordance with the invention.
[0266] Each of the foregoing kits can further include written
instructions directing an end user how to assemble and/or
administer the various components.
[0267] Each of the foregoing kits can further include an applicator
or other delivery device suitably constructed and arranged for use
in administering the various and/or assembled components to a
subject in need thereof. For example, in certain embodiments such
applicator or delivery device can be a syringe.
[0268] The invention may be further understood by reference to the
following nonlimiting examples.
EXAMPLES
Materials
[0269] Silk fibroin was obtained from Silk lab, Tufts Science and
Technology Center, 4 Colby St Biomed Eng Dept., Medford, Mass.
[0270] PFC was obtained from Origen Biomedical, Austin, Tex.
[0271] Lung Surfactant called Infasurf was obtained from ONY, Inc.
1576 Sweet Home Road, Amherst, N.Y. 14228.
Example 1: Preparation and Characterization of Mixtures of Silk
Fibroin, PFC, and Surfactant
[0272] Aqueous preparations of silk fibroin and perfluorocarbon
(PFC); silk fibroin and surfactant; PFC and surfactant; and silk
fibroin, PFC, and surfactant were prepared and visually inspected
for homogeneity. Selected properties of silk fibroin and PFC are
shown in Table 1. Selected properties of silk fibroin and
surfactant are shown in Table 2. Representative results are shown
in FIGS. 1A-1E.
TABLE-US-00001 TABLE 1 Selected properties of silk fibroin and PFC
Property Silk Fibroin PFC Drug delivery effect + + pH 6-6.5 5-6
O.sub.2 permeability + +++ Biodegradability +++ +++
Biocompatibility +++ +++ Water solubility Yes No
TABLE-US-00002 TABLE 2 Selected properties of silk fibroin and
surfactant Property Silk Fibroin Surfactant Constituents protein
and water protein and lipid Proteins Hydrophobic (HMW) Hydrophobic
(BC+) Hydrophilic (LMW) Hydrophilic (A-) Surface Tension ?? Low
(14-18 Dynes/cm) H.sub.2O: 70 Dynes/cm) Lung: 25 Dynes/cm pH 6-6.5
5.0-6.2
[0273] Silk fibroin and surfactant formed a homogeneous mixture.
Surfactant and PFC mixture was nonhomogeneous. Silk fibroin, PFC,
and surfactant was homogeneous after vigorous shaking.
Example 2: Silk Fibroin/Perflubron Administration in
Surfactant-Deficient Animal Lungs (Rabbit Model)
[0274] A New Zealand white rabbit was used to test proof of
concept. The animal weighing 1.58 kg anesthetized and after
instrumentation, anesthesia was administered continuously as per
protocol. The rabbit was intubated orally and ventilated on
assist/control mode at 30 breaths/min, 0.4 fraction of inspired
oxygen (FiO.sub.2), 0.35 sec inspiratory time, 4 cm H.sub.2O
positive end expiratory pressure (PEEP), and tidal volume targeted
to 7 mL/kg. The lungs were lavaged 4 times with 30 mL/kg of normal
(0.9%) saline several times until blood gases were consistent with
surfactant deficiency status. After a stabilization period of 30
min on the ventilator, arterial blood gas (ABG) analyses, mean
arterial pressure (MAP), and arterial blood pressures were measured
prior to administration of liquid fibroin and PFC 1:2 volume ratio
(1 mL of silk fibroin to 2 mL of PFC). Oxygen saturations, arterial
blood gases, and blood pressure were recorded at the end.
Representative results are shown in Table 3.
TABLE-US-00003 TABLE 3 Effect of administration of silk fibroin/PFC
to surfactant-deficient rabbit. Before After Parameter Fibroin/PFC
Fibroin/PFC O.sub.2 saturation 85% 92% ABG 7.20/65/45/20/-9
7.31/55/60/23/-5 pH/PaCO.sub.2/PaO.sub.2/HCO.sub.3/base excess BP
(mm Hg) 35/19 39/23 MAP (mm Hg) 24 28
[0275] Administration of mixture of liquid silk fibroin and PFC to
surfactant-deficient rabbit resulted in improvement in oxygen
saturations and arterial blood gases and stabilization of mean
blood pressure. Surfactant has already been shown to be effective
in surfactant deficient-lung, strongly suggesting that inclusion of
surfactant would only further improve results in this system.
[0276] The observed improvement in blood gases and blood pressure
after instillation of mixture of silk fibroin and PFC shows
possible synergy between two liquids on surfactant-deficient lungs
and suggests such mixture will have beneficial effect upon its
addition to surfactant and administration to surfactant-deficient
lungs.
Example 3. Head-to-Head Comparison of Silk Fibroin/PFC/Surfactant
to Individual Components and Pairs of Components
[0277] Rats, rabbits, or sheep are anesthetized, intubated,
ventilated, and made surfactant-deficient in a manner similar to
Example 2. Animals are then treated with test agent selected from
(i) silk fibroin alone; (ii) PFC alone; (iii) surfactant alone;
(iv) vehicle alone; (v) silk fibroin and PFC; (vi) silk fibroin and
surfactant; (vii) PFC and surfactant; and/or (viii) silk fibroin,
PFC, and surfactant by instillation into the lungs. Arterial blood
gas (ABG), mean arterial pressure (MAP), and arterial blood
pressures are measured prior to administration of test agent.
Oxygen saturations, arterial blood gases, and blood pressures are
recorded after administration of test agent. Comparison is made
between pre- and post-administration clinical parameters. Results
are expected to show that (viii) silk fibroin, PFC, and surfactant
is superior to (i) silk fibroin alone; (ii) PFC alone; (iii)
surfactant alone; (iv) vehicle alone; (v) silk fibroin and PFC;
(vi) silk fibroin and surfactant; and (vii) PFC and surfactant,
including synergistic improvement over (v) silk fibroin and PFC;
(vi) silk fibroin and surfactant; and/or (vii) PFC and surfactant.
Sequence CWU 1
1
116PRTBombyx mori 1Gly Ser Gly Ala Gly Ala1 5
* * * * *