U.S. patent application number 11/827362 was filed with the patent office on 2008-01-31 for compositions and methods for the treatment of cystic fibrosis.
Invention is credited to Douglas V. Faller, Susan P. Perrine, George Stamatoyannopoulos.
Application Number | 20080027136 11/827362 |
Document ID | / |
Family ID | 22118869 |
Filed Date | 2008-01-31 |
United States Patent
Application |
20080027136 |
Kind Code |
A1 |
Faller; Douglas V. ; et
al. |
January 31, 2008 |
Compositions and methods for the treatment of cystic fibrosis
Abstract
The invention is directed to novel pharmaceutical compositions
comprising chemicals agents that are useful in the treatment and
prevention of cystic fibrosis and the prevention of signs and
symptoms of this disease. These pharmaceutical compositions are
surprisingly successful in the treatment disorders related to
cystic fibrosis including disorders of blood production. Many of
these compositions of the invention are even more effective when
administered to a patient in pulses. Pulse therapy is not a form of
discontinuous administration of the same amount of a composition
over time, but comprises administration of the same dose of the
composition at a reduced frequency or administration of reduced
doses.
Inventors: |
Faller; Douglas V.; (Weston,
MA) ; Perrine; Susan P.; (Weston, MA) ;
Stamatoyannopoulos; George; (Seattle, WA) |
Correspondence
Address: |
RONALD I. EISENSTEIN;NIXON PEABODY LLP
100 SUMMER STREET
BOSTON
MA
02110
US
|
Family ID: |
22118869 |
Appl. No.: |
11/827362 |
Filed: |
July 11, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10185745 |
Jul 1, 2002 |
7265153 |
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11827362 |
Jul 11, 2007 |
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09248260 |
Feb 11, 1999 |
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10185745 |
Jul 1, 2002 |
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60074304 |
Feb 11, 1998 |
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Current U.S.
Class: |
514/529 ;
514/557; 514/568; 514/574 |
Current CPC
Class: |
A61K 31/216 20130101;
A61K 31/165 20130101; A61K 31/19 20130101; A61K 31/191 20130101;
A61K 31/215 20130101; A61K 31/194 20130101; A61P 1/18 20180101;
A61K 31/00 20130101; A61K 31/195 20130101; A61P 5/48 20180101; A61K
31/22 20130101; A61P 7/00 20180101; A61K 31/192 20130101; A61P
11/00 20180101; A61P 1/00 20180101 |
Class at
Publication: |
514/529 ;
514/557; 514/568; 514/574 |
International
Class: |
A61K 31/215 20060101
A61K031/215; A61K 31/19 20060101 A61K031/19; A61P 1/00 20060101
A61P001/00 |
Claims
1. A method for the treatment or prevention of cystic fibrosis
comprising the administration of a composition comprising a
physiologically-effective amount of one or more agents selected
from the group consisting of butyric acid ethyl ester, 2,2-dimethyl
butyric acid, 2,2-diethyl butyric acid, 3,3-dimethyl butyric acid,
3,3-diethyl butyric acid, 2,3-dimethyl succinic acid, methoxy
acetic acid, phenoxyacetic acid, 2- and 3-thiophenoxy propionic
acid, 2- and 3-phenoxy propionic acid, 2- and 3-phenyl propionic
acid, 4-chlorophenoxy-2-propionic acid, methoxy acetic acid, or
2-thiophenoxy acetic acid, or a chemical compound of the structure
phenyl-R.sub.9--R.sub.10 wherein R.sub.9 is CH.sub.x, CO, NH.sub.x,
OX.sub.x, SH.sub.x, or a branched or linear aryl chain; R.sub.10 is
CH.sub.x, CO, H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x, CONH.sub.x,
COOH, COSH.sub.x, COOR.sub.x, COR.sub.x, CO or OR.sub.11; and
R.sub.11 is CH.sub.x, CO, H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x or
a branched or linear alkyl chain; wherein x is 0, 1, 2 or 3.
2. The method of claim 1 wherein the chemical compound of the
structure phenyl-R9-R10 is selected from the group consisting of
acids, amines and amides of cinnamic acid, hydrocinnamic acid,
dihydrocinnamic acid, a-methyl hydrocinnamic acid, dihydro cinnamic
acid, 2,3-dimethyl hydrocinnamic, dihydrocinnamic acid, phenyl
acetate ethyl ester, 2-phenoxypropionic acid, phenoxy acetic acid,
or 3-phenyl butyric acid.
3. The method of claim 1 wherein the one or more agents is
substituted with one or more halogens.
4. The method of claim 3 wherein the halogen is selected from the
group consisting of chlorine, fluorine, iodine, bromine or mixtures
or combinations thereof.
5. The method of claim 1 wherein administration is pulsed
administration or timed-release administration.
6. The method of claim 5 wherein the pulsed administration
comprises a plurality of individual pulses delivered to a patient
continuously over a period of 2 hours, 4 hours, 6 hours, 8 hours,
10 hours, 12 hours, 14 hours 16 hours, 2 days, 3 days, 4 days, 5
days, 6 days, 7 days, two weeks, three weeks or four weeks.
7. The method of claim 5 wherein the pulsed administration
comprises a plurality of individual pulses delivered at regular
intervals measuring from between 3 to 9 hours.
8. The method of claim 1 wherein the composition further comprises
a pharmaceutically acceptable carrier.
9. The method of claim 1 wherein the composition further comprises
a compound that positively affects expression of a CFTR
molecule.
10. The method of claim 9 wherein the compound that positively
affects expression of the CFTR increases the extent or magnitude of
CFTR function, increases the expression of the CFTR molecule,
increases transport of the CFTR molecule to the cell surface,
increases half-life of the CFTR molecule, increases expression from
a CFTR gene, increases CFTR transcript levels, increases
post-transcriptional processes which increases CFTR transcript
levels in the cell, or increases translation post-translational
processing of a CFTR gene product.
11. The method of claim 1 wherein the agent treats defective
chloride ion transport.
12. A method for the therapy of cystic fibrosis comprising
administering to a patient a quantity of an agent, or
pharmaceutically acceptable derivatives thereof, effective for said
therapy, said agent selected from the group consisting of butyric
acid ethyl ester, 2,2-dimethyl butyric acid, 2,2-diethyl butyric
acid, 3,3-dimethyl butyric acid, 3,3-diethyl butyric acid,
2,3-dimethyl succinic acid, methoxy acetic acid, phenoxyacetic
acid, 2- and 3-thiophenoxy propionic acid, 2- and 3-phenoxy
propionic acid, 2- and 3-phenyl propionic acid,
4-chlorophenoxy-2-propionic acid, methoxy acetic acid,
2-thiophenoxy acetic acid, or a chemical compound f the structure
phenyl-R.sub.9--R.sub.10 wherein R.sub.9 is CH.sub.x, CO, NH.sub.x,
OH.sub.x, SH.sub.x, or a branched or linear aryl chain; R.sub.10 is
CH.sub.x, CO, H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x, CONH.sub.x,
COOH, COSH.sub.x, COOR.sub.11, COR.sub.11, CO or OR.sub.11; and
R.sub.11 is CH.sub.x, CO, H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x or
a branched or linear alkyl chain; wherein x is 0, 1, 2 or 3.
13. The method of claim 12 wherein the chemical compound of the
structure phenyl-R9-R10 is selected from the group consisting of
acids, amines and amides of cinnamic acid, hydrocinnamic acid,
dihydriocinnamic acid, a-methyl hydrocinnamic acid, dihydro
cinnamic acid, 2,3-dimethyl hydrocinnamic, dihydrocinnamic acid,
phenyl acetate ethyl ester, 2-phenoxypropionic acid, phenoxy acetic
acid, and 3-phenyl butyric acid.
14. A method for enhancing expression of CFTR comprising the
administration of a physiologically effective amount of one or more
agents or pharmaceutically acceptable derivatives thereof, said
agents selected from the group consisting of butyric acid ethyl
ester, 2,2-dimethyl butyric acid, 2,2-diethyl butyric acid,
3,3-dimethyl butyric acid, 3,3-diethyl butyric acid, 2,3-dimethyl
succinic acid, methoxy acetic acid, phenoxyacetic acid, 2- and
3-thiophenoxy propionic acid, 2- and 3-phenoxy propionic acid, 2-
and 3-phenyl propionic acid, 4-chlorophenoxy-2-propionic acid,
methoxy acetic acid, 2-thiophenoxy acetic acid, and chemical
compounds of the structure phenyl-R.sub.9--R.sub.10 wherein R.sub.9
is CH.sub.x, CO, NH.sub.x, OH.sub.x, SH.sub.x, or a branched or
linear aryl chain; R.sub.10 is CH.sub.x, CO, H.sub.x, NH.sub.x,
OH.sub.x, SH.sub.xCONH.sub.x, COOH, COSH.sub.x, COOR.sub.11,
COR.sub.11, CO or OR.sub.11; and R.sub.11 is CH.sub.x, CO, H.sub.x,
NH.sub.x, OH.sub.x, SH.sub.x or a branched or linear alkyl chain;
wherein x is 0, 1, 2 or 3.
15. The method of claim 14 wherein the chemical compounds of the
structure phenyl-R.sub.9--R.sub.10 are selected from the group
consisting of acids, amines and amides of cinnamic acid,
hydrocynnamic acid, dihydrocinnamic acid, a-methyl hydrocinnamic
acid, dihydro cinnamic acid, 2,3-dimethyl hydrocinnamic,
dihydrocinnamic acid, phenyl acetate ethyl ester,
2-phenoxypropionic acid, phenoxy acetic acid, and 3-phenyl butyric
acid.
16. The method of claim 14 wherein administration is pulsed
administration.
17. The method of claim 14 wherein enhancement of the expression of
CFTR comprises increasing the expression of CFTR genes, increasing
the number of CFTR-expressing cells or increasing the function or
activity of CFTR.
18. The method of claim 14 wherein CFTR expression is enhanced
greater than about 30%.
19. The method of claim 14 wherein CFTR expression is enhanced
greater than about 100%.
20. The method of claim 14 wherein CFTR expression is enhanced
greater than about 200%.
Description
FIELD OF THE INVENTION
[0001] The invention relates to pharmaceutically acceptable
compositions for administration to humans to treat cystic fibrosis
and also to methods for effectively utilizing these
compositions.
BACKGROUND OF THE INVENTION
[0002] Cystic fibrosis (CF) is a systemic disorder that results
when mutations in the cystic fibrosis transmembrane conductance
regulator (CFTR), an apical membrane glycoprotein, lead to a
reduction in apical membrane chloride transport. CFTR is a
cAMP-dependent chloride channel that regulates fluid composition in
the respiratory and gastrointestinal tracts. CF is a heritable
disease that follows an autosomal recessive pattern of
transmission. It is the most common invariably lethal genetic
disease in the United States, with frequency among Caucasians being
one in two thousand. One in twenty are carriers of the defective
gene. CF is characterized by abnormal endocrine and exocrine gland
function. In CF, unusually thick mucus leads chronic pulmonary
disease and respiratory infections, insufficient pancreatic and
digestive function, and abnormally concentrated sweat. Seventy
percent of the mutant CFTR alleles in the Caucasian population
result from deletion of phenylalanine at position 508 (AF508-CFTR),
the result of a three base pair deletion in the genetic code. Other
mutations have also been described and many may exist. The
.DELTA.F508-CFTR mutation results in a CFTR protein capable of
conducting chloride, but absent from the plasma membrane because of
aberrant intracellular processing. Under usual conditions
(37.degree. C.), the .DELTA.F508-CFTR protein is retained in the
endoplasmic reticulum (ER), by prolonged association with the ER
chaperones, including calnexin and hsp70. The retained CFTR protein
is then targeted for degradation by the ubiquitin proteasome
pathway. Over expression of .DELTA.F508-CFTR can result in
.DELTA.F508-CFTR protein appearing at the cell surface, and this
protein is functional once it reaches the cell surface. The
.DELTA.F508 "trafficking" block is also reversible by incubation of
cultured CF epithelial cells at reduced temperatures (25-27.degree.
C.). Lowered temperature results in the appearance of CFTR protein
and channel activity at the cell surface, suggesting an intrinsic
thermodynamic instability in .DELTA.F508-CFTR at 37.degree. C. that
leads to recognition of the mutant protein by the ER quality
control mechanism, prevents further trafficking, and results in
protein degradation. High concentrations of glycerol (1 M or 10%),
a protein stabilizing agent or chemical chaperone, also appears to
facilitate movement of .DELTA.F508-CFTR from the ER to the plasma
membrane.
[0003] Some of the palliative treatments involve the administration
of biologically active proteins or chemical compounds to decrease
the viscosity of secretions, or to suppress chronic infections of
the airways. These treatments have a number of limitations, and do
not address the illness directly, but rather attempt to treat the
symptoms. Some require continuous use at fairly high doses while
others have short effective half-lives. Tolerance to the active
ingredient often develops rendering the composition functionally
useless. In addition to problems associated with tolerance, the
substances themselves or their metabolic by-products or carriers
can quickly reach toxic levels in the patient's system which impair
kidney or liver function. Further, the chemical compounds
themselves can be rapidly destroyed by catabolic enzymes, found in
the cells and serum such as aminases, oxidases and hydrolases. Many
of these enzymes are also found in hepatic cells, the principal
sites for cleansing of the blood. Those able to survive cellular
and hepatic catabolic processes are quickly eliminated from the
patient's system by the kidneys. Consequently, in vivo retention
times for active compounds are extremely short and the ability to
achieve any sort of sustained biological effect becomes nearly
impossible or, at least, impractical.
[0004] Gene therapy for cystic fibrosis has been attempted, but has
not been successful to date for a number of reasons, including
problems with delivery of the gene to airway cells, insufficient
levels of gene expression, inadequate duration of gene expression,
and toxicity of the gene therapy preparations.
[0005] A recent publication used 4-phenylbutyrate (4PBA) to enable
a greater fraction of .DELTA.F508-CFTR to escape degradation and
appear at the cell surface (Rubenstein, R. C., Egan, M. E., and
Zeitlin, P. L. In vitro pharmacologic restoration of CFTR-mediated
chloride transport with sodium 4-phenyl butyrate in cystic fibrosis
epithelial cells containing delta-F508-CFTR. J. Clin. Invest.
100:2457-65, 1997). Briefly, primary cultures of nasal polyp
epithelia from CF patients (.DELTA.F508 homozygous or
heterozygous), or the CF bronchial epithelial cell line IB3-1
(.DELTA.F508/W1282X) were exposed to 4PBA for up to 7 days in
culture. 4PBA treatment at concentrations of 0.1 and 2 mM resulted
in the restoration of forskolin-activated chloride secretion.
Protein kinase A-activated, linear, 10 pS chloride channels
appeared at the plasma membrane of IB3-1 cells at the tested
concentration of 2.5 mM 4PBA. Treatment of IB3-1 cells with 0.1-1
mM 4PBA and primary nasal epithelia with 5 mM 4PBA also resulted in
the appearance of higher molecular mass forms of CFTR, consistent
with addition and modification of oligosaccharides in the Golgi
apparatus, as detected by immunoblotting of whole cell lysates with
anti-CFTR antisera. Immunocytochemistry in CF epithelial cells
treated with 4PBA was consistent with increasing amounts of
.DELTA.F508-CFTR.
[0006] As 4PBA is an analogue of butyrate, a known transcriptional
regulator of CFTR expression (Cheng, S. H., Fang, S. L., Zabner,
J., Marshall, J., Piraino, S., Schiavi, S. C., Jefferson, D. M.,
Welsh, M. J., and Smith, A. E. Functional activation of the cystic
fibrosis trafficking mutant .DELTA.F508-CFTR by expression. Am. J.
Physiol. 268:L615-24, 1995), it was hypothesized that 4PBA might
increase transcription of the .DELTA.F508-CFTR allele (Rubenstein
et al.). If it were a transcriptional regulator, 4PBA might thereby
increase levels of .DELTA.F508-CFTR protein, and by mass action,
would force some .DELTA.F508-CFTR to bypass quality control in the
ER. Such a mechanism would be consistent with the observations that
butyrate itself can induce cAMP-responsive chloride secretion in a
.DELTA.F508-homozygous pancreatic acinar cell line (Cheng et al.).
The results observed were consistent with 4PBA increasing the
amount of .DELTA.F508-CFTR protein produced, but their data
demonstrated that this was not due to a transcriptional regulatory
effect of 4PBA on the CFTR gene. In immunoblot experiments,
increased CFTR immunoreactivity was observed in the 4PBA-treated
samples. Increased CFTR immunoreactivity was also observed by
immunocytochemistry after 4PBA treatment, but no changes in CFTF
RNA levels were found with 4PBA treatment. The authors further
stated that butyrate and 4PBA have effects in IB3-1 cells that are
qualitatively different from one another. Respiratory epithelial
cells treated with 1-2 mM 4PBA are healthy, grow at a similar rate
and with a similar morphology to control cells, and express CFTR
channel activity at the plasma membrane. Equimolar concentrations
of butyrate caused morphologic changes in IB3-1 cells, with
rounding of cells and decreased growth rate.
[0007] This seems to indicate that 4PBA and butyrate may have
different toxicity profiles and dose-response relationships. In
addition, other published observations with butyrate in
.DELTA.F508-CFTR transfected C-127 cells found that the
.about.180-kD mature glycosylated species of CFTR was not observed
after 5 mM butyrate treatment for 24 hours, despite a massive
increase in .DELTA.F508-CFTR mRNA as demonstrated by Northern
analysis (Cheng et al.). This data thus did not demonstrate any
effects of butyrate on CFTR protein levels or function, only
changes in cellular morphology and cell death (Rubenstein et al.).
Rubenstein et al observed no increases in CFTR mRNA in response to
4PBA and indicated that the mechanism of action of 4PBA was not
similar to that of butyrate or related to increasing
.DELTA.F508-CFTR transcription. In addition, no increases in
cAMP-stimulation was observed which would be indicative of chloride
ion transport even after treatment with up to 300 mM butyrate
(Cheng et al.).
[0008] These data argue against any beneficial or therapeutic
effect of butyrate on cystic fibrosis. In fact, some authors even
stated that butyrate is likely too toxic to use clinically
(Rubenstein et al.). Further, the authors made a strong case that
4PBA, which was indicated to be possibly clinically useful, works
though a mechanism, which although unknown, is different from
butyrate. Taken together, the use of butyrate, and the newer
butyrate-derived compounds claimed, as CF therapeutics is
contra-indicated according to these reports. Moreover, 4PBA has
been used in a few CF patients clinically, but was not well
tolerated due to large number of pills required (i.e. very short
half-life), and other side effects and, in consideration, that
study was terminated.
DESCRIPTION OF THE INVENTION
[0009] As embodied and broadly described herein, the present
invention is directed to novel chemicals and novel pharmaceutical
compositions comprising these and other chemicals that can be used
in the treatment and prevention of diseases and disorders
associated with cystic fibrosis. The invention is further directed
to methods for the administration of these pharmaceutical
compositions to patients for the treatment of cystic fibrosis and
prevention of its signs and symptoms.
[0010] It has been discovered that a group of chemicals and
pharmaceutical compositions containing one or more such chemicals
are surprisingly successful in the treatment of cystic fibrosis and
other disorders including, for example, disorders of blood
production. Also surprisingly, it was discovered that many of these
compositions are even more effective when administered to a patient
in pulses. Pulse therapy is not a form of discontinuous
administration of the same amount of a composition over time, but
comprises administration of the same dose of the composition at a
reduced frequency or administration of reduced doses.
[0011] According to these methods, cystic fibrosis and other
disorders can be effectively treated and without unnecessary
adverse side effects to the patient. Although most compositions are
generally safe and non-toxic at therapeutic doses, pulsed
administration further reduces risks associated with, for example,
toxicity, allergic reactions, the build-up of toxic metabolites and
inconveniences associated with conventional treatment. In addition,
these chemical compositions, now useful at a substantially reduced
dose and frequency, have a significantly reduced risk of
complications such as, for example, induced tolerance. These
compositions are not inactivated by cellular enzymes or cleared
from cells and organs prior to having the desired effect. Further,
long-term therapy, typically required for the amelioration of many
blood disorders, can be successfully performed. Consequently, doses
necessary for maintaining a constant effect for the patient are
steady and material costs and inconveniences associated with
administration are substantially reduced.
[0012] The mechanism of action of many of the chemical compounds or
active ingredients of compositions for the treatment of cystic
fibrosis involves effecting one or more of the processes of gene
transcription, protein translation or processing or transport or
stability, cell proliferation, cell recruitment, cell
differentiation, or CFTR expression or activity. Gene expression
can be increased or decreased by altering chromatin and/or
nucleosome structure to render a genetic element more or less
susceptible to transcription, by altering DNA structure, for
example, by methylation of G residues, by affecting the activity of
cell-specific transcription or translation factors such as
activators or repressors, or by increasing the rate of
transcription or translation. CFTR expression can be increased or
decreased by affecting gene expression, peptide expression, CFTR
assembly, CFTR glycosylation or transport through the Golgi
apparatus or the stability of the CFTR molecule. Cell proliferation
may be increased, for example, by stimulating stem cells, pulmonary
or pancreatic or other secretory cell growth, or decreased, for
example, by effecting a cell's period in or ability to transverse a
stage (S, G2, G 1, M) of the cell cycle. Cell recruitment may be
promoted through the expression of specific cytokines such as cell
surface receptors or secreted factors. CFTR function may be
increased by promoting chloride transport or other activities of
the protein.
[0013] Chemical agents that can be administered as pharmaceutical
compositions include phenoxyacetic acid, methoxyacetic acid,
butyric acid ethyl ester, cinnamic acid, hydrocinnamic acid,
alpha-methyl cinnamic acid and alpha-methylhydrocinnamic acid
(alpha-MHCA) which stimulate alterations in binding or removal of
transcription factors from the proximal promoter region of certain
genes or gene clusters and thereby increase suppressed gene
expression, or serve a chaperones to facilitate processing,
transport and the thermal or physical stability of mutated or
normal CFTR proteins.
[0014] These compositions preferably increase the expression of
CFTR, increase the expression of CFTR genes, increase the number of
CFTR-expressing cells or increase the activity of CFTR. Preferably,
compositions also increase CFTR expression or function greater than
about 30%, more preferably greater than about 100%, and even more
preferably greater than about 200%. CFTR intracellular and cell
surface expression, gene expression and cell proliferation can be
assayed by measuring fold increases in expressed amounts of
specific mRNA, protein or numbers of CFTR-expressing cells in
treated samples as compared untreated controls. Utilizing this
criteria, compositions preferably increase the amount of CFTR cell
surface expression, the amount of CFTR gene expression, the number
of CFTR-expressing cells by greater than or equal to about
11/2-fold, preferably about two-fold and more preferably about
four-fold. CFTR function can be measured by analysis of chloride
ion transport/efflux (cAMP-stimulated or otherwise), patch
clamping, sweat testing, or improvement in the symptoms of cystic
fibrosis.
[0015] One embodiment of the invention is directed to
pharmaceutical compositions comprising one or more novel chemical
agents. Agents include chemicals of the structure
R.sub.1--R.sub.2--R.sub.3 or, preferably,
R.sub.1--C(O)--R.sub.2--R.sub.3 wherein R.sub.1 is CH.sub.x, CO,
H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x, COH.sub.x, CONH.sub.x, COOH
or COSH.sub.x; R.sub.2 is CH.sub.x or a branched or linear alkyl
chain; R.sub.3 is CONH.sub.x, COSH.sub.x, COOH, COOR.sub.4,
COR.sub.4, CO or OR.sub.4; R.sub.4 is CH.sub.x, CO, H.sub.x,
NH.sub.x, OH.sub.x, SH.sub.x or a branched or linear alkyl chain;
phenyl-R.sub.5--R.sub.6--R.sub.7 wherein phenyl is a six carbon
benzyl ring or a hydrogenated, hydroxylated or halogenated six
carbon ring; R.sub.5 is CH.sub.x, CO, NH.sub.x, OH.sub.x or
SH.sub.x: R.sub.6 is CH.sub.x, CO, H.sub.x, NH.sub.x, OH.sub.x,
SH.sub.x or a branched or linear alkyl chain; R.sub.7 is CH.sub.x,
H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x, CO, CONH.sub.x, COOH,
COSH.sub.x, COOR.sub.8, COR.sub.8 or OR.sub.8; R.sub.8 is CH.sub.x,
CO, H.sub.x, NH.sub.x, OH.sub.x, SH.sub.x or a branched or linear
aryl chain; and phenyl-R.sub.9--R.sub.10 wherein R.sub.9 is
CH.sub.x, CO, NH.sub.x, OH.sub.x, SH.sub.x, or a branched or linear
aryl chain; R.sub.10 is CH.sub.x, CO, H.sub.x, NH.sub.x, OH.sub.x,
SH.sub.x, CONH.sub.x, COOH, COSH.sub.x, COOR.sub.11, COR.sub.11, CO
or OR.sub.11; and R.sub.11 is CH.sub.x, CO, H.sub.x, NH.sub.x,
OH.sub.x, SH.sub.x or a branched or linear alkyl chain; wherein x
is 0, 1, 2 or 3. Preferably, R.sub.4 comprises between 1 to 8
carbon atoms and more preferably 1, 2, 3 or 4 carbon atoms.
Preferably, R.sub.6 comprises between 1 to 8 carbon atoms and more
preferably 1, 2, 3 or 4 carbon atoms. Preferably, R.sub.8 comprises
between 1 to 8 carbon atoms and more preferably 1, 2, 3 or 4 carbon
atoms.
[0016] Examples of chemical compounds of the structure
R.sub.1--R.sub.2--R.sub.3 or R.sub.1--C(O)--R.sub.2-R.sub.3 include
acids, amines, monoamides and diamides of butyric acid
(H.sub.3C--CH.sub.2--CH.sub.2--COOH), butyric acid ethyl ester
(CH.sub.2CH.sub.2CH.sub.2COCH.sub.2CH).sub.3 4,4,4-tri
fluorobutyric acid (CF.sub.3CH.sub.2CH.sub.2COOH), 2,2-dimethyl
butyric acid (C.sub.2H.sub.5C(CH.sub.3).sub.2CO.sub.2H),
2,2-diethyl butyric acid, 3,3-dimethyl butyric acid
(C.sub.6H.sub.12O.sub.2), 3,3-diethyl butyric acid, f umaric acid
(HOOCCH.dbd.CHCOOH), flumaric acid monomethyl and monoethyl ester,
fumaric acid monoamide (C.sub.4H.sub.5O.sub.2N), fumaramide
(H.sub.2NCOCCHCONH.sub.2), succinic acid (HOOCCH.sub.2CH.sub.2COOH)
(succinamic acid and succinamide), 2,3-dimethyl succinic acid and
methoxy acetic acid (CH.sub.3CH.sub.2OCH.sub.3).
[0017] Examples of chemical compounds of the structure
phenyl-R.sub.5--R.sub.6--R.sub.7 include acids, amines and amides
of phenoxyacetic acid (C.sub.6H.sub.5OCH.sub.2COOH; C.sub.6H.sub.5
OCH.sub.2COONH.sub.3), 2- and 3-thiophenoxy propionic acid
(C.sub.6H.sub.5SCH(CH.sub.3)COOH; C.sub.6H.sub.5
SCH.sub.2CH.sub.2COOH), 2- and 3-phenoxy propionic acid
(C.sub.6H.sub.5OCH(CH.sub.3)COOH; C.sub.6H.sub.5OCH.sub.2
CH.sub.2COOH), 2- and 3-phenyl propionic acid
(C.sub.6H.sub.5CH(CH.sub.3)COOH; C.sub.6H.sub.5CH.sub.2CH.sub.2
COOH), 4-chlorophenoxy-2-propionic acid
(ClC.sub.6OCH.sub.2CH.sub.2CO.sub.2H), methoxy acetic acid
(H.sub.3COCH.sub.2CO.sub.2H), and 2-thiophenoxy acetic acid
(C.sub.6H.sub.5SCH.sub.2COOH).
[0018] Examples of chemical compounds of the structure
phenyl-R.sub.9--R.sub.10 include acids, amines and amides of
cinnamic acid (C.sub.6H.sub.5CH.dbd.CHCOOH), hydrocinnamic acid,
dihydrocinnamic acid (C.sub.6H.sub.5CH.sub.2CH.sub.2COOH), a-methyl
hydrocinnamic acid or dihydro cinnamic acid, 2,3-dimethyl
hydrocinnamic or dihydrocinnamic acid, phenyl acetate ethyl ester
(C.sub.6H.sub.5CH(CH.sub.3)CH.sub.2COCH.sub.2CH.sub.3),
2-phenoxypropionic acid (C.sub.6H.sub.5OCH.sub.2CO.sub.2H), phenoxy
acetic acid (CH.sub.3CH(OC.sub.6H.sub.5)CO.sub.2H, and 3-phenyl
butyric acid (C.sub.6H.sub.3CH(CH.sub.3)CH.sub.2COOH). Additional
chemical compounds which may or may not be included in the above
classification scheme include monobutyrin, tributyrin
(CH.sub.2(OCOCH.sub.2CH.sub.2CH.sub.3)CH(OCOCH.sub.2CH.sub.2CH.sub.3)CH.s-
ub.2(OCOCH.sub.2CH.sub.2CH.sub.3), ethyl-phenyl acetic acid
(CH.sub.3 CH.sub.2C.sub.6H.sub.5CH.sub.2COOH), indol-3-propionic
acid, indol-3-butyric acid, 1- and 2-methyl cyclopropane carboxylic
acid (C.sub.5H.sub.8O.sub.2 and C.sub.6H.sub.8O.sub.2),
mercaptoacetic acid (C.sub.2H.sub.4O.sub.2S), N-acetylglycine
(C.sub.4H.sub.7O.sub.3N), squaric acid (C.sub.4H.sub.2O.sub.4),
4-trifluorobutanol (C.sub.4H.sub.7OF.sub.3), chloropropionic acid
(ClCH.sub.2CH.sub.2CO.sub.2H), 3-trimethyl silyl-1-proposulfonic
acid sodium (C.sub.6H.sub.15O.sub.3SS), 2-oxopantansane
(C.sub.5H.sub.8O.sub.3), isobutyl hydroxyl amine HCl
(C.sub.4H.sub.12OCl), 2-methyl butanoic acid
(C.sub.5H.sub.10O.sub.2), o-benzoyl lactate, n-dimethylbutyric acid
glycine amide, o-dimethyl butyric acid lactate, and diethyl butyric
acid.
[0019] Agents are useful in pharmaceutical compositions for the
treatment of cystic fibrosis. Preferred agents in such compositions
include, for example, propionic acid, butyric acid, succinic acid,
fumaric acid monoethyl ester, dimethyl butyric acid,
trifluorobutanol (C.sub.4H.sub.7OF.sub.3), chloropropionic acid
(ClCH.sub.2CH.sub.2COOH), isopropionic acid, 2-oxypentasane
(CH.sub.3CH.sub.2CH.sub.2C(O)COOH), 2,2- or 3,3-dimethyl butyric
acid (C.sub.6H.sub.12O.sub.2), 2,2- or 3,3-diethyl butyric acid
(C.sub.8H.sub.16O.sub.2), butyric acid ethyl ester, 2-methyl
butanoic acid (C.sub.5H.sub.10O.sub.2), fumaric acid
(C.sub.4H.sub.4O.sub.3) and amides and salts thereof. Other
examples include methoxy acetic acid (H.sub.3C(O)CH.sub.2COOH),
dimethyl butyric acid, methoxy propionic acid, N-acetylglycine
(H.sub.3CC(O)NCH.sub.2COOH), mercaptoacetic acid (HSCH.sub.2 COOH),
1- or 2-methyl cyclopropane carboxylic acid
(C.sub.5H.sub.8O.sub.2), squaric acid (C.sub.4H.sub.2O.sub.4), 2-
or 3-phenoxy propionic acid, methoxy butyric acid, phenoxy acetic
acid, 4-chloro-2-phenoxy 2-propionic acid, 2- or 3-phenoxy butyric
acid, phenyl acetic acid, phenyl propionic acid, 3-phenyl butyric
acid, ethyl-phenyl acetic acid, 4-chloro-2-phenoxy-2-propionic
acid, n-dimethyl butyric acid glycine amide, o-benzoyl lactic acid,
o-dimethyl butyric acid lactate, cinnamic acid, dihydrocinnamic
acid (C.sub.6H.sub.5CHCH.sub.3COOH), a-methyl-dihydrocinnamic acid,
thiophenoxy acetic acid, and amines, amides and salts of these
chemicals.
[0020] Useful amines and amides include isobutylhydroxylamine:HCl
(C.sub.4H.sub.12OCl), fumaric acid monoamide
(C.sub.4H.sub.5O.sub.2N), fumaramide (H.sub.2NCOCHCHCONH.sub.2),
succinamide and isobutyramide (C.sub.4H.sub.9ON). Salts can be
sodium, potassium, calcium, ammonium, lithium or choline such as
sodium 3-trimethyl silyl-1-proposulfonic acid
(C.sub.6H.sub.15O.sub.3SiS:Na). Reagents which may be
electrostatically or covalently bonded with the inducing agent
include amino acids such as arginine (arginine butyrate), glycine,
alanine, asparagine, glutamine, histidine or lysine, nucleic acids
including nucleosides or nucleotides, or substituents such as
carbohydrates, saccharides, lipids, fatty acids, proteins or
protein fragments. Combinations of these salts with the inducing
agent can also produce useful new compounds from the interaction of
the combination.
[0021] Chemical compounds are preferably optically pure with a
specific conformation (plus {+} or minus {-}), absolute
configuration (R or S), or relative configuration (D or L).
Particular salts such as sodium, potassium, magnesium, calcium,
choline, amino acid, ammonium or lithium, or combinations of salts
may also be preferred, however, certain salts may be more
advantageous than others. For example, chemical compositions that
require high doses may introduce too much of a single salt to the
patient. Sodium is generally an undesirable salt because at high
doses, sodium can increase fluid retention resulting in tissue
destruction. In such instances, lower doses or combinations of
different or alternative salts can be used. For example, compounds
of the invention may be substituted with one or more halogens such
as chlorine (Cl), fluorine (F), iodine (I), bromine (Br) or
combinations of these halogens. As known to those of ordinary skill
in the art, halogenation can increase the polarity, hydrophilicity
or lipophilicity or a chemical compound which can be a desirable
feature, for example, to transform a chemical compound into a
composition which is more easily tolerated by the patient or more
readily absorbed by the epithelial lining of the gastrointestinal
tract. Such compositions could be orally administered to
patients.
[0022] Therapeutically effective chemical compounds may be created
by modifying any of the above chemical compounds so that after
introduction into the patient, these compounds metabolize into
active forms, such as the forms above, which have the desired
effect on the patient. Compounds may also be created which are
metabolized in a timed-release fashion allowing for a minimal
number of introductions which are efficacious for longer periods of
time. Combinations of chemical compounds can also produce useful
new compounds from the interaction of the combination. Such
compounds may also produce a synergistic effect when used in
combination with other known or other compounds.
[0023] Compositions are preferably physiologically stable at
therapeutically effective concentrations. Physiological stable
compounds are compounds that do not break down or otherwise become
ineffective upon introduction to a patient prior to having a
desired effect. Compounds are structurally resistant to catabolism,
and thus, physiologically stable, or coupled by electrostatic or
covalent bonds to specific reagents to increase physiological
stability. Such reagents include ammo acids such as arginine,
glycine, alanine, asparagine, glutamine, histidine or lysine,
nucleic acids including nucleosides or nucleotides, or substituents
such as carbohydrates, saccharides and polysaccharides, lipids,
fatty acids, proteins, or protein fragments. Useful coupling
partners include, for example, glycol such as polyethylene glycol,
glucose, glycerol, glycerin and other related substances.
[0024] Physiological stability can be measured from a number of
parameters such as the half-life of the compound or the half-life
of active metabolic products derived from the compound. Certain
compounds of the invention have in vivo half lives of greater than
about fifteen minutes, preferably greater than about one hour, more
preferably greater than about two hours, and even more preferably
greater than about four hours, eight hours, twelve hours or longer.
Although a compound is stable using this criteria, physiological
stability cam also be measured by observing the duration of
biological effects on the patient. Clinical symptoms which are
important from the patient's perspective include a reduced
frequency or duration, or elimination of the need for oxygen,
inhaled medicines, or pulmonary therapy. Preferably, a stable
compound of the invention has an in vivo half-life of greater than
about 15 minutes, a serum half-life of greater than about 15
minutes, or a biological effect which continues for greater than 15
minutes after treatment has been terminated or the serum level of
the compound has decreased by more than half.
[0025] Preferably, compositions are also not significantly
biotransformed, degraded or excreted by catabolic processes
associated with metabolism. Although there may be some
biotransformation, degradation or excretion, these functions are
not significant if the composition is able to exert its desired
effect.
[0026] Compositions are also preferably safe at effective dosages.
Safe compositions are compositions that are not substantially toxic
(e.g. cytotoxic or myelotoxic), or mutagenic at required dosages,
do not cause adverse reactions or side effects, and are
well-tolerated. Although side effects may occur, compositions are
substantially safe if the benefits achieved from their use outweigh
disadvantages that may be attributable to side effects. Unwanted
side effects include nausea, vomiting, hepatic or renal damage or
failure, hypersensitivity, allergic reactions, cardiovascular
problems, gastrointestinal disturbances, seizures and other central
nervous system difficulties, fever, bleeding or hemorrhaging, serum
abnormalities and respiratory difficulties.
[0027] Compositions useful for treating disorders preferably do not
substantially affect the viability of a cell such as a normal
mammalian cell, the cell being treated or effected by the chemical
compound. Normal cell viability, the viability of an untransformed
or uninfected cell, can be determined from analyzing the effects of
the composition on one or more biological processes of the cell.
Detrimental interference with one or more of these cellular
processes becomes significant when the process becomes abnormal.
Examples of quantitatable and qualifiable biological processes
include the processes of cell division, protein synthesis, nucleic
acid (DNA or RNA) synthesis, nucleic acid (principally DNA)
fragmentation and apoptosis. Others processes include specific
enzyme activities, the activities of the cellular transportation
systems such as the transportation of amino acids by system A
(neutral), system B (acidic) or system C (basic), and the
expression of a cell surface protein. Each of these parameters is
easily determined as significantly detrimental, for example, in
tissue culture experiments, in animal experiments or in clinical
studies using techniques known to those of ordinary skill in the
art. Abnormal cell division, for example, can be mitosis which
occurs too rapidly, as in a malignancy, or unstably, resulting in
programmed cell death or apoptosis, detected by increased DNA
degradation. The determination of abnormal cell viability can be
made on comparison with untreated control cells. Compositions
preferably increase normal cell viability. Increased cell viability
can be determined by those of ordinary skill in the art using, for
example, DNA fragmentation analysis. A decreased amount of
fragmentation indicates that cellular viability is boosted.
Determinations of increased or decreased viability can also be
concluded from an analysis of the results of multiple different
assays. Where multiple tests provide conflicting results, accurate
conclusions can still be drawn by those of ordinary skill based
upon the cell type, the correctness or correlation of the tests
with actual conditions and the type of composition.
[0028] Compositions can be prepared in solution as a dispersion,
mixture, liquid, spray, capsule or as a dry solid such as a powder
or pill, as appropriate or desired. Solid forms may be processed
into tablets or capsules or mixed or dissolved with a liquid such
as water, alcohol, saline or other salt solutions, glycerol,
saccharides or polysaccharide, oil or a relatively inert solid or
liquid. Liquids, pills, capsules or tablets administered orally may
also include flavoring agents to increase palatability.
Additionally, all compositions may further comprise agents to
increase shelf-life, such as preservatives, anti-oxidants and other
components necessary and suitable for manufacture and distribution
of the composition. Compositions further comprise a
pharmaceutically acceptable carrier. Carriers are chemical or
multi-chemical compounds that do not significantly a lter or effect
the active ingredients of the compositions. Examples include water,
alcohols such as glycerol and polyethylene glycol, glycerin, oils,
salts such as sodium, potassium, magnesium and ammonium, fatty
acids, saccharides or polysaccharides. Carriers may be single
substances or chemical or physical combinations of these
substances.
[0029] Another embodiment of the invention is directed to
combinations of compositions comprising a chemical compound in
combination with an agent known to positively affect expression of
the CFTR molecule. The agent may be a chemical compound such as
glycerol, acetic acid, butyric acid, D- or L-amino-n-butyric acid,
alpha- or beta-amino-n-butyric acid, arginine butyrate or
isobutyramide, all disclosed in U.S. Pat. Nos. 4,822,821 and
5,025,029. Others include butyrin, 4-phenyl butyrate
(C.sub.6H.sub.5CH.sub.2CH.sub.2CH.sub.2COOH), phenylacetate
(C.sub.6H.sub.5CH.sub.2COOH), phenoxy acetic acid, all of which and
more are disclosed in U.S. Pat. No. 4,704,402, and U.S. patent
application Ser. No. 08/398,588 (entitled "Compositions for the
Treatment of Blood Disorders" filed Mar. 3, 1995), and derivatives,
salts and combination of these agents. The agent may be a protein
such as hsp70 or a growth factor or cytokine. The agent may be a
gene or a nucleotide sequence. Such composition may have additive
or synergistic effects.
[0030] In another embodiment, compositions of the invention may
contain one or more chemical compounds that increase the extent or
magnitude of CFTR function, increase the expression of the CFTR
molecule, increase transport of the CFTR molecule to the cell
surface, increase the half-life (physical stability or thermal
stability) of the molecule, increase expression from the CFTR gene,
increase CFTR transcript levels, or increase post-transcriptional
processes which increase the levels of CFTR transcript, or increase
translation or enhance post-translational processing of the CFTR
gene product. Stimulation of specific gene expression involves
activation of transcription or translation promoters or enhancers,
or alteration of the methylation patterns or histone distribution
along the gene to promote expression. Expression may also be
stimulated by inhibition of specific transcriptional or
translational repressors, activation of specific transcriptional or
translational activation factors, or activation of receptors on the
surface of particular populations of cells. Stimulation may recruit
additional epithelial cells to the airways, reprogram
differentiated epithelial cells to express CFTR. Stimulation may
also activate a previously dormant or relatively inactive gene.
[0031] Compositions of the invention may be administered by oral,
parenteral, sublingual, rectal or enteral administration, or
pulmonary absorption or topical application. Compositions cam be
directly or indirectly administered to the patient. Indirect
administration is performed, for example, by administering the
composition to cells ex vivo and subsequently introducing the
treated cells to the patient. The cells may be obtained from the
patient to be treated or from a genetically related or unrelated
patient. Related patients offer some advantage by lowering the
immunogenic response to the cells to be introduced. For example,
using techniques of antigen matching, immunologically compatible
donors can be identified and utilized.
[0032] Direct administration of a composition may be by oral,
parenteral, sublingual, rectal such as suppository or enteral
administration, or by pulmonary absorption or topical application.
Parenteral administration may be by intravenous injection,
subcutaneous injection, intramuscular injection, intra-arterial
injection, intrathecal injection, intraperitoneal injection or
direct injection or other administration to the desired site.
Injectable forms of administration are sometimes preferred for
maximal effect. When long term administration by injection is
necessary medi-ports, in-dwelling catheters, or automatic pumping
mechanisms are also preferred wherein direct and immediate access
is provided to the arteries in and around the heart and other major
organs and organ systems.
[0033] An effective method of administration to a specific site may
be by transdermal transfusion such as with a transdermal patch, by
direct contact to the cells or tissue, if accessible, such as a
skin tumor, or by administration to an internal site through an
incisions or some other artificial opening into the body.
Compositions may also be administered to the nasal passages as a
spray. Diseases localized to the head and brain area are treatable
in this fashion as arteries of the nasal area provide a rapid and
efficient access to the upper areas of the head. Sprays also
provide immediate access to the pulmonary system and are the
preferable methods for administering compositions to these areas.
Access to the gastrointestinal tract is gained using oral, enema,
or injectable forms of administration. Compositions may be
administered as a bolus injection or spray, or administered
sequentially over time (episodically) such as every two, four, six
or eight hours, every day (QD) or every other day (QOD), or over
longer periods of time such as weeks to months.
[0034] Orally active compositions are preferred, as oral
administration is usually the safest, most convenient and
economical mode of drug delivery. Oral administration is usually
disadvantageous because compositions are poorly absorbed through
the gastrointestinal lining. Compounds which are poorly absorbed
tend to be highly polar. Consequently, compounds which are
effective, as described herein, may be made orally bioavailable by
reducing or eliminating their polarity. This can often be
accomplished by formulating a composition with a complimentary
reagent which neutralizes its polarity, or modifying the compound
with a neutralizing chemical group. Oral bioavailability is also a
problem because drugs are exposed to the extremes of gastric pH and
gastric enzymes. These problems can be overcome in a similar matter
by modifying the molecular structure to be able to withstand very
low pH conditions and resist the enzymes of the gastric mucosa such
as by neutralizing an ionic group, by covalently bonding an ionic
interaction, or by stabilizing or removing a disulfide bond or
other relatively labile bond.
[0035] Compounds may also be used in combination with other agents
to maximize the effect of the compositions in an additive or
synergistic manner. Cytokines which may be effective in combination
with the compositions of the invention include growth factors such
as B cell growth factor (BCGF), fibroblast-derived growth factor
(FGF), granulocyte/macrophage colony stimulating factor (GM-CSF),
granulocyte colony stimulating factor (G-CSF), macrophage colony
stimulating factor (M-CSF), epidermal growth factor (EGF), vascular
endothelial growth factor (VEGF), platelet derived growth factor
(PDGF) nerve growth factor (NGF), stem cell factor (SCF), and
transforming growth factor (TGF). These growth factors plus a
composition may further stimulate cellular differentiation and/or
the expression of the CFTR molecule or function.
[0036] Alternatively, other cytokines and related antigens in
combination with a composition may also be useful to treat cystic
fibrosis. Potentially useful cytokines include tumor necrosis
factor (TNF), the interleukins IL-1, IL-2, IL-3, IL-4, IL-5, IL-6,
etc., recombinant IL receptors, growth factors, colony stimulating
factors, erythropoietin (EPO), the interferon (IFN) proteins
IFN-alpha, IFN-beta, and IFN-gamma; cyclic AMP including dibutyryl
cyclic AMP, hemin, DMSO, hydroxyurea, hypoxanthine, glucocorticoid
hormones and cytosine arabinoside. Therapies using combinations of
these agents would be safe and effective therapies cystic fibrosis.
Combinations of therapies may also be effective in inducing
improvement of the symptoms of cystic fibrosis such as compositions
of the invention plus the reintroduction of a normal or altered
CFTR gene (gene therapy), toxin or drug conjugated antibody therapy
using monoclonal or polyclonal antibodies directed against the
pulmonary cells, or specific anti-sense therapy. Effects may be
additive, logarithmic or synergistic, and methods involving
combinations of therapies may be simultaneous protocols,
intermittent protocols or protocols which are empirically
determined.
[0037] Another embodiment of the invention is directed to the
pulsed administration of pharmaceutical compositions for the
treatment or prevention of cystic fibrosis. Pulsed administration
is surprisingly more effective than continuous treatment as pulsed
doses are often lower than would be expected from continuous
administration of the same composition. Each pulse dose can be
reduced and the total amount of drug administered over the course
of treatment to the patient is minimized.
[0038] In traditional forms of therapy, repeated administration is
designed to maintain a desired level of an active ingredient in the
body. Very often, complications that develop can be attributed to
dosage levels that, to be effective, are near toxic or otherwise
harmful to normal cells. In contrast, with pulse therapy, in vivo
levels of drug drop below that level required for effective
continuous treatment. Therefore, pulsing is not simply the
administration of a sufficiently large bolus such that there will
be therapeutically sufficient drug available for a long period of
time. Pulsed administration can substantially reduce the amount of
the composition administered to the patient per dose or per total
treatment regimen with an increased effectiveness. This represents
a significant saving in time, effort and expense and, more
importantly, a lower effective dose substantially lessens the
number and severity of complications that may be experienced by the
patients. As such, pulsing is surprisingly more effective than
continuous administration of the same composition.
[0039] Preferably, compositions contain chemicals that are
substantially non-toxic. Substantially non-toxic means that the
composition, although possibly possessing some degree of toxicity,
is not harmful to the long-term health of the patient. Although the
active component of the composition may not be toxic at required
levels, there may also be problems associated with administering
the necessary volume or amount of the final form of the composition
to the patient. For example, if the composition contains a salt,
although the active ingredient may be at a concentration that is
safe and effective, there can be a harmful build-up of sodium,
potassium or another ion. With a reduced requirement for the
composition or at least the active component of that composition,
the likelihood of such problems can be reduced or even eliminated.
Consequently, although patients may have minor or short term
detrimental side-effects, the advantages of taking the composition
outweigh the negative consequences.
[0040] Compositions most effective at pulsed administration are
typically non-toxic or non-cytotoxic chemicals without any
substantial proteinaceous active component at the therapeutically
effective pulsed dose. Preferably, treatment does not stimulate
apoptosis in the cells being directly treated or in the otherwise
normal cells of the body which will also be exposed to the
composition.
[0041] Individual pulses can be delivered to the patient
continuously over a period of several hours, such as about 2, 4, 6,
8, 10, 12, 14 or 16 hours, or several days, such as 2, 3, 4, 5, 6,
or 7 days, preferably from about 1 hour to about 24 hours and more
preferably from about 3 hours to about 9 hours. Alternatively,
periodic doses can be administered in a single bolus or a small
number of injections of the composition over a short period of
time, typically less than 1 or 2 hours. For example, arginine
butyrate has been administered over a period of 4 days with
infusions for about 8 hours per day or overnight, followed by a
period of 7 days of no treatment. The interval between pulses or
the interval of no delivery is greater than 24 hours and preferably
greater than 48 hours, and can be for even longer such as for 3, 4,
5, 6, 7, 8, 9 or 10 days, two, three or four weeks or even longer.
As the results achieved may be surprising, the interval between
pulses, when necessary, can be determined by one of ordinary skill
in the art. Often, the interval between pulses can be calculated by
administering another dose of the composition when the composition
or the active component of the composition is no longer detectable
in the patient prior to delivery of the next pulse. Intervals can
also be calculated from the in vivo half-life of the composition.
Intervals may be calculated as greater than the in vivo half-life,
or 2, 3, 4, 5 and even 10 times greater the composition half-life.
For compositions with fairly rapid half lives such as arginine
butyrate with a half-life of 15 minutes, intervals may be 25, 50,
100, 150, 200, 250 300 and even 500 times the half life of the
chemical composition.
[0042] The number of pulses in a single therapeutic regimen may be
as little as two, but is typically from about 5 to 10, 10 to 20, 15
to 30 or more. In fact, patients can receive drugs for life
according to the methods of this invention without the problems and
inconveniences associated with current therapies. Compositions can
be administered by most any means, but are preferably delivered to
the patient as an injection (e.g. intravenous, subcutaneous,
intraarterial), infusion or instillation, and more preferably by
oral ingestion. Various methods and apparatus for pulsing
compositions by infusion or other forms of delivery to the patient
are disclosed in U.S. Pat. Nos. 4,747,825; 4,723,958; 4,948,592;
4,965,251 and 5,403,590.
[0043] Compositions administered in pulses have the surprising
benefit of reducing the overall load of drug on the patient as the
total amount of drug administered can be substantially less than
that amount that has been therapeutically administered by
conventional continuous therapy. Substantially means that there is
more than an insignificant difference between the amount or
concentration of a composition administered by pulsing according to
the invention verses the amount or concentration administered using
conventional therapy, without compromising the beneficial effect
achieved to the patient. For example, arginine butyrate has been
shown to be effective at continuous administration at about 2000
mg/kg patient weight. Doses of between about 400 to 1500 mg/kg,
preferably from about 600 to 1000 mg/kg and more preferably from
700 to 800 mg/kg, when administered in pulses, are surprisingly
more beneficial as measured by a rise in fetal hemoglobin levels in
thalassemic patients. Typical pulsed amounts of arginine butyrate
are from about 2 to about 20 g/kg/month, and preferably from about
3 to about 10 g/kg/month wherein the patient receives a total of
less than about 20 kg per month, preferably less than about 15 kg
per month and more preferably less than about 10 kg per month. The
amounts administered per pulse as well as the total amount of the
composition received by the patient over the regimen is
substantially reduced. Preferably, the therapeutically effective
pulsed dose is less than the continuous dose, or less than one
half, one third, one quarter, one fifth, one tenth or even one
twentieth of the therapeutic continuous dose of the same
composition or even less.
[0044] Other embodiments and uses of the invention will be apparent
to those skilled in the art from consideration of the specification
and practice of the invention disclosed herein. All U.S. patents
and patent applications, including provisional applications, and
all other documents referenced herein, for whatever reason, are
specifically incorporated by reference. It is intended that the
specification and examples be considered exemplary only, with the
true scope and spirit of the invention being indicated by the
following claims.
* * * * *