U.S. patent application number 12/397806 was filed with the patent office on 2009-07-30 for method for augmenting the effects of serotonin reuptake inhibitors.
This patent application is currently assigned to Duke University. Invention is credited to Martin Beaulieu, Marc G. Caron, Raul R. Gainetdinova, Ranga R. Krishnan, Tatiana D. Sotnikova, Xiaodong Zhang.
Application Number | 20090192166 12/397806 |
Document ID | / |
Family ID | 35428824 |
Filed Date | 2009-07-30 |
United States Patent
Application |
20090192166 |
Kind Code |
A1 |
Krishnan; Ranga R. ; et
al. |
July 30, 2009 |
METHOD FOR AUGMENTING THE EFFECTS OF SEROTONIN REUPTAKE
INHIBITORS
Abstract
A method of treating a subject for a serotonergic
neurotransmission dysregulation disorder, comprises administering
the subject a serotonin enhancer (e.g., a serotonin reuptake
inhibitor) in an amount effective to treat the disorder; and
concurrently administering the subject 5-hydroxytryptophan in an
amount effective to enhance the activity of the serotonin enhancer,
(e.g., serotonin reuptake inhibitor). In preferred embodiments the
disorder is depression, anxiety, or substance abuse.
Inventors: |
Krishnan; Ranga R.; (Chapel
Hill, NC) ; Caron; Marc G.; (Hillsborough, NC)
; Zhang; Xiaodong; (Durham, NC) ; Beaulieu;
Martin; (Durham, NC) ; Gainetdinova; Raul R.;
(Chapel Hill, NC) ; Sotnikova; Tatiana D.; (Chapel
Hill, NC) |
Correspondence
Address: |
MYERS BIGEL SIBLEY & SAJOVEC
PO BOX 37428
RALEIGH
NC
27627
US
|
Assignee: |
Duke University
Durham
NC
|
Family ID: |
35428824 |
Appl. No.: |
12/397806 |
Filed: |
March 4, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11133867 |
May 20, 2005 |
7517908 |
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12397806 |
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60642869 |
Jan 11, 2005 |
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60642800 |
Jan 11, 2005 |
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60629951 |
Nov 22, 2004 |
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60606811 |
Sep 2, 2004 |
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60573265 |
May 21, 2004 |
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Current U.S.
Class: |
514/249 ;
514/338; 514/419; 514/469; 514/567; 514/640; 514/647; 514/651 |
Current CPC
Class: |
A61K 45/06 20130101;
C12Q 2600/106 20130101; A61K 31/343 20130101; A61K 31/405 20130101;
A61K 31/14 20130101; C12Q 1/6883 20130101; A61K 31/137 20130101;
A61P 25/24 20180101; C12Q 2600/156 20130101 |
Class at
Publication: |
514/249 ;
514/338; 514/419; 514/469; 514/567; 514/640; 514/647; 514/651 |
International
Class: |
A61K 31/405 20060101
A61K031/405; A61K 31/519 20060101 A61K031/519; A61K 31/4525
20060101 A61K031/4525; A61K 31/343 20060101 A61K031/343; A61K
31/195 20060101 A61K031/195; A61K 31/15 20060101 A61K031/15; A61K
31/136 20060101 A61K031/136; A61K 31/138 20060101 A61K031/138 |
Goverment Interests
GOVERNMENT FUNDING
[0003] This invention was made with Government support under grant
numbers 5 P50 MII60451 and 5 P50 MH60451 from the National
Institute of Mental Health. The United States Government has
certain rights to this invention.
Claims
1. A method of treating a subject for a serotonergic
neurotransmission dysregulation disorder, comprising: determining
the presence or absence of a tryptophan hydroxylase 2 (Tph2)
mutation in said subject; and then, if said subject possesses said
Tph2 mutation; administering said subject a serotonin reuptake
inhibitor in an amount effective to treat said disorder; and
concurrently administering said subject 5-hydroxytryptophan in an
amount effective to enhance the activity of said serotonin reuptake
inhibitor; to treat said serotonergic neurotransmission
deregulation disorder.
2. The method of claim 1, wherein said subject is a human
subject.
3. The method of claim 2, wherein said Tph2 is human Tph2 and said
mutation encodes a change in an amino acid of the encoded protein,
said amino acid selected from the group consisting of A65, V66,
F68, L77, F84, I94, R97, E105, P152, W153, P155, D162, L175, R191,
E211, V223, P244, G251, R276, P277, V278, R285, R294, P308, Y310,
E313, A333, 1339, A342, S343, L344, A346, K353, V421, E423, A428,
A436, R441, Y446, P449, Y450, and Q468.
4. The method of claim 2, wherein said Tph2 mutation is selected
from the group consisting of the human P449R mutation, the human
R441H mutation, the human WI 53R mutation, the human A65V mutation,
the human V66I mutation, the human L175V mutation, and the human
Q468X (where X is a stop codon) mutation.
5. The method of claim 1, wherein said serotonin reuptake inhibitor
is selected from the group consisting of citalopram, fluvoxamine,
fluoxetine, sertraline, paroxetine, escitalopram, duloxetine,
venlafaxine, and pharmaceutically acceptable salts thereof.
6. The method of claim 1, wherein said disorder is depression.
7. The method of claim 1, wherein said disorder is anxiety.
8. The method of claim 1, wherein said disorder is substance
abuse.
9. The method of claim 1, further comprising the step of
concurrently administering said subject a peripheral decarboxylase
inhibitor in an amount effective to reduce peripheral degradation
of said 5-hydroxytryptophan.
10. The method of claim 9, wherein said decarboxylase inhibitor is
selected from the group consisting of carbidopa and
benserazide.
11.-73. (canceled)
74. The method of claim 1, further comprising the step of
concurrently administering said subject tetrahydrobiopterin.
75. The method of claim 1, wherein said disorder is
attention-deficit/hyperactivity disorder (ADHD).
76. The method of claim 1, wherein said disorder is obsessive
compulsive disorder.
77. The method of claim 1, wherein said disorder is bipolar
disorder.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 11/133,867, filed May 20, 2005, and also
claims the benefit of U.S. Provisional Patent Application Ser. No.
60/642,869 filed Jan. 11, 2005, and of U.S. Provisional Patent
Application Ser. No. 60/642,800, filed Jan. 11, 2005, and of U.S.
Provisional Patent Application Ser. No. 60/629,951, filed Nov. 22,
2004, and of U.S. Provisional Patent Application Ser. No.
60/606,811, filed Sep. 2, 2004, and of U.S. Provisional Patent
Application Ser. No. 60/573,265, filed May 21, 2004, the
disclosures of all of which are incorporated by reference herein in
their entirety.
[0002] This application is related to Marc G. Caron, Xiaodong
Zhang, Martin Beaulieu, Raul R. Gainetdinov, Tatiana D. Sotnikova,
Ranga R. Krishnan, David A. Schwartz, Lauranell Burch, and Redford
B. Williams, Polymorphism in Tryptophan Hydroxylase-2 Controls
Brain Serotonin Synthesis, U.S. patent application Ser. No.
11/133,949, filed May 20, 2005, and published as US 2006/0029951,
the disclosure of which is incorporated by reference herein in its
entirety.
FIELD OF THE INVENTION
[0004] The present invention relates to methods for the treatment
of serotonin neurotransmission dysregulation disorders, including
but not limited to depression, anxiety and substance abuse.
BACKGROUND OF THE INVENTION
[0005] Brain serotonin system plays a critical role in numerous
neuronal functions and dysregulation of its homeostasis may
contribute to many psychiatric disorders (Veenestra-VanderWeele et
al, 2000; Flattem and Blakely 2000; Blier and Abbott 2001; Tecott,
2003; Geyer, 1996; Tamming a, 1998; Snyder and Peroutka, 1982;
Frazer, 1997; Aghajanian and Marek, 2000). In fact, numerous
conditions such as unipolar major depression and bipolar disorder,
obsessive-compulsive disorder, anxiety, autism, personality
disorder, panic and eating disorders, suicidality, chronic pain and
post-traumatic stress syndrome and even attention deficit
hyperactivity disorder (ADHD), are effectively treated by raising
the extracellular concentrations of serotonin in the brain with
compounds including selective serotonin reuptake inhibitors (SSRIs)
that inhibit the neuronal re-uptake of serotonin. Like many other
neurotransmitters and modulators, the actions of serotonin are
modulated by the delicate balance between synthesis and degradation
of this monoamine. Tryptophan hydroxylase-1 (Tph1) has long been
considered as the sole rate-limiting enzyme for the synthesis of
serotonin. However, Walther et al (2003) recently reported that
inactivation of the Tph1 gene in the mouse led to a decrease in the
peripheral levels of serotonin but no changes in central serotonin
levels, suggesting that another form of the enzyme might exist.
Data base mining yielded a second related gene that was identified,
cloned and named TPH2. The product of the TPH2 gene is
preferentially expressed in the brain as opposed to the predominant
peripheral expression pattern of TPH1 (Walther et al, 2003).
[0006] M. Bader et al., PCT Application WO 2004/007704 (US
2006/0275759), describes the identification of TPH2.
SUMMARY OF THE INVENTION
[0007] A first aspect of the present invention is a method of
treating a subject for a serotonergic neurotransmission
dysregulation disorder, comprising: administering the subject a
serotonin enhancer (e.g., a serotonin reuptake inhibitor) in an
amount effective to treat the disorder; and concurrently
administering the subject 5-hydroxytryptophan in an amount
effective to enhance the activity of the serotonin enhancer, (e.g.,
serotonin reuptake inhibitor). In preferred embodiments the
disorder is depression, anxiety, or substance abuse.
[0008] A second aspect of the present invention is a method of
treating a subject for a serotonergic neurotransmission
dysregulation disorder, comprising: administering the subject a
serotonin enhancer (e.g., a serotonin reuptake inhibitor) in an
amount effective to treat the disorder; and concurrently
administering the subject a peripheral decarboxylase inhibitor such
as carbidopa in an amount effective to enhance the activity of the
serotonin enhancer (e.g., selective serotonin reuptake
inhibitor).
[0009] A third aspect of the invention is a method of treating a
subject for a serotonergic neurotransmission dysregulation
disorder, comprising: administering the subject a serotonin
enhancer (e.g., a serotonin reuptake inhibitor) in an amount
effective to treat said disorder; and concurrently administering
the subject a BH4 enhancer such as tetrahydrobiopterin in an amount
effective to enhance the activity of the serotonin enhancer (e.g.,
serotonin reuptake inhibitor).
[0010] A fourth aspect of the present invention is a method of
treating a subject for a serotonergic neurotransmission
dysregulation disorder, comprising: determining the presence or
absence of at least one Tph2 mutation in the subject; and then, if
the subject possesses at least one Tph2 mutation; administering the
subject 5-hydroxytryptophan in an amount effective to treat the
disorder.
[0011] A fifth aspect of the present invention is a method of
treating a subject for a serotonergic neurotransmission
dysregulation disorder, comprising: determining the presence or
absence of at least one Tph2 mutation in the subject; and then, if
the subject possesses at least one Tph2 mutation; administering the
subject a peripheral decarboxylase inhibitor in an amount effective
to treat the disorder.
[0012] A sixth aspect of the present invention is a method of
treating a subject for a serotonergic neurotransmission
dysregulation disorder, comprising: determining the presence or
absence of at least one Tph2 mutation in said subject; and then, if
said subject possesses at least one Tph2 mutation; administering
said subject a BH4 enhancer such as tetrahydrobiopterin in an
amount effective to treat said disorder.
[0013] In some embodiments of the foregoing, the disorder is
depression, anxiety, or substance abuse; in other embodiments the
disorder is another serotonergic neurotransmission dysregulation
disorder such as described below.
[0014] A further aspect of the present invention is a
pharmaceutical formulation comprising, consisting of, or consisting
essentially of:
[0015] (a) a serotonin enhancer such as a serotonin reuptake
inhibitor;
[0016] (b) a serotonin precursor such as 5-hydroxytryptophan;
[0017] (c) optionally, but in some embodiments preferably, a
peripheral decarboxylase inhibitor such as carbidopa;
[0018] (d) optionally, but in some embodiments preferably, a BH4
enhancer such as tetrahydrobiopterin; and
[0019] (e) a pharmaceutically acceptable carrier.
[0020] A further aspect of the present invention is a
pharmaceutical formulation comprising, consisting of, or consisting
essentially of:
[0021] (a) a serotonin enhancer such as a serotonin reuptake
inhibitor;
[0022] (b) a peripheral decarboxylase inhibitor such as
carbidopa;
[0023] (c) optionally, but in some embodiments preferably, a BH4
enhancer such as tetrahydrobiopterin;
[0024] (d) optionally, but in some embodiments preferably, a
serotonin precursor such as 5-hydroxytryptophan; and
[0025] (e) a pharmaceutically acceptable carrier.
[0026] A further aspect of the present invention is a
pharmaceutical formulation comprising, consisting of, or consisting
essentially of:
[0027] (a) a serotonin enhancer such as a serotonin reuptake
inhibitor;
[0028] (b) a BH4 enhancer such as tetrahydrobiopterin;
[0029] (c) optionally, but in some embodiments preferably, a
peripheral decarboxylase inhibitor such as carbidopa;
[0030] (d) optionally, but in some embodiments preferably, a
serotonin precursor such as 5-hydroxytryptophan; and
[0031] (e) a pharmaceutically acceptable carrier.
[0032] A further aspect of the present invention is a
pharmaceutical formulation comprising, consisting of, or consisting
essentially of:
[0033] (a) a serotonin precursor such as 5-hydroxytryptophan;
[0034] (b) a peripheral decarboxylase inhibitor such as
carbidopa;
[0035] (c) optionally, but in some embodiments preferably, a BH4
enhancer such as tetrahydrobiopterin; and
[0036] (d) a pharmaceutically acceptable carrier.
[0037] A further aspect of the present invention is a
pharmaceutical or neutraceutical formulation comprising, consisting
of, or consisting essentially of:
[0038] (a) a serotonin precursor such as 5-hydroxytryptophan;
and
[0039] (b) a BH4 enhancer such as tetrahydrobiopterin; and
[0040] (c) a pharmaceutically acceptable carrier; or in the case of
a neutraceutical optionally other food ingredients such as fats,
proteins, and/or carbohydrates.
[0041] A further aspect of the present invention is the use of
5-hydroxytryptophan for the preparation of a medicament for
carrying out a method of as described herein.
[0042] A further aspect of the present invention is the use of a
peripheral decarboxylase inhibitor such as carbidopa for the
preparation of a medicament for carrying out a method as described
herein.
[0043] A further aspect of the present invention is the use of a
BH4 enhancer such as tetrahydrobiopterin for the preparation of a
medicament for carrying out a method as described herein.
[0044] A further aspect of the present invention is the use of a
serotonin enhancer such as a serotonin reuptake inhibitor for the
preparation of a medicament for carrying out a method as described
herein.
[0045] A still further aspect of the present invention is the use
of a means for detecting a Tph2 mutation for carrying out a method
as described herein.
[0046] The foregoing and other objects and aspects of the present
invention are explained in greater detail in the drawings herein
and the specification set forth below.
BRIEF DESCRIPTION OF THE DRAWINGS
[0047] FIG. 1. Serotonin levels in PC12 cells expressing R441H
hTPH2 were lower than those in PC12 cells expressing WT hTPH2 (n=8
experiments).
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0048] The present invention is explained in greater detail below.
This description is not intended to be a detailed catalog of all
the different ways in which the invention may be implemented, or
all the features that may be added to the instant invention. For
example, features illustrated with respect to one embodiment may be
incorporated into other embodiments, and features illustrated with
respect to a particular embodiment may be deleted from that
embodiment. In addition, numerous variations and additions to the
various embodiments suggested herein will be apparent to those
skilled in the art in light of the instant disclosure which do not
depart from the instant invention. Hence, the following
specification is intended to illustrate some particular embodiments
of the invention, and not to exhaustively specify all permutations,
combinations and variations thereof.
[0049] The disclosures of all United States Patent references cited
herein are to be incorporated by reference herein in their
entirety.
1. DEFINITIONS
[0050] "Genotyping" as used herein means determination of the type
and number of alleles present in a subject, whether determined by
nucleic acid sequencing, PCR or RT-PCR amplification, examination
of Tph2 protein, or any other method. A specific gene can be
genotyped to determine if the gene is a wild-type or variant
allele. Genotyping does not require sequencing of the entire gene
but may simply involve determining the presence or absence of one
or more mutations therein, as compared to the "wild type" gene.
[0051] "Tph2 mutation" as used herein refers to any mutation which
leads to loss or decrease in activity of the encoded protein
(tryptophan hydroxylase-2), such that the conversion of tryptophan
to 5-hydroxytryptophan in the subject by tryptophan hydroxylase-2
is reduced.
[0052] "Serotonin enhancer" as used herein refers to any compound
that increases, directly or indirectly, the availability of
serotonin in the central nervous system for binding to serotonin
receptors at the post-synaptic membrane, including but not limited
to serotonin reuptake inhibitors, monoamine oxidase inhibitors,
tricyclic antidepressants, serotonin agonists, amphetamines,
serotonin precursors, serotonin prodrugs, intermediates in the
biosynthesis of serotonin, co-factors and pharmaceutically
acceptable salts thereof. Such compounds may be given alone or in
combination with other serotonin enhancers.
[0053] "Serotonergic neurotransmission dysregulation disorder" as
used herein refers to any disorder in which an increase or decrease
in available serotonin contributes, at least in part, to a disease,
disorder, or condition. Examples of such disorders include, but are
not limited to, depressive disorder, anxiety disorder, social
anxiety disorder, generalized anxiety disorder, bipolar disorder,
schizophrenia, autism, epilepsy, mood disorders, alcohol or
substance abuse and associated disorders, panic disorder, migraine,
obesity, bulimia, anorexia, premenstrual syndrome, menopause, sleep
disorders, attention-deficit/hyperactivity disorder (ADHD),
Tourette syndrome, aggression, obsessive compulsive disorder,
pathological gambling, novelty seeking, borderline personality
disorders, antisocial personality disorder, suicidality, eating
disorders, sexual dysfunction, dementia, social phobia,
fibromyalgia, overactive bladder, chronic fatigue syndrome, chronic
pain, sudden infant death syndrome, post-traumatic stress syndrome,
and Alzheimer's disease. These terms have their usual meaning in
the art (see, e.g., DSM-IV).
[0054] "Depression" as used herein has its usual meaning in the art
and includes treating subjects with symptoms of depression and
treating subjects diagnosed as depressed (e.g., subjects diagnosed
as afflicted with unipolar major depression)
[0055] "Unipolar major depression" (also called "unipolar
depression" or "major depression") as used herein has its usual
meaning in the art (see, e.g., DSM-IV), and is typically
characterized by the presence of at least five of the following
symptoms for at least two weeks: Trouble sleeping or excessive
sleeping; a dramatic change in appetite, often with weight gain or
loss; fatigue and lack of energy; feelings of worthlessness,
self-hate, and inappropriate guilt; extreme difficulty
concentrating; agitation, restlessness, and irritability;
inactivity and withdrawal from usual activities; feelings of
hopelessness and helplessness; recurring thoughts of death or
suicide.
[0056] "Anxiety" as used herein has its conventional meaning in the
art and includes the treatment of subjects exhibiting symptoms of
anxiety.
[0057] "Substance abuse" as used herein has its usual meaning and
includes both alcohol abuse or addiction (e.g., alcoholism, or
alcoholic subjects), as well as abuse or addiction to drugs such as
narcotics, opiates, stimulants, depressants, etc. (e.g.,
barbiturates, ecstasy, cocaine, crack cocaine, morphine, heroin,
amphetamine, methamphetamine, oxycontin, etc.).
[0058] Subjects for screening and/or treatment with the present
invention are, in general, human subjects, but may also be other
mammalian subjects (e.g., rodent subjects such as mouse or rat,
primate subjects such as human or monkey, dog, cat, rabbit, etc.)
for veterinary purposes. Subjects includes male and female
subjects. The subject may be of any race and any age, including
juvenile, adolescent, and adult.
[0059] "Treating" as used herein means the medical management of a
subject, e.g., a human patient, with the intent to cure,
ameliorate, stabilize, or prevent a disease, pathological
condition, or disorder. This term includes active treatment, that
is, treatment directed specifically toward the improvement or
associated with the cure of a disease, pathological condition, or
disorder, and also includes causal treatment, that is, treatment
directed toward removal of the cause of the associated disease,
pathological condition, or disorder. In addition, this term
includes palliative treatment, that is, treatment designed for the
relief of symptoms rather than the curing of the disease,
pathological condition, or disorder; preventative treatment, that
is, treatment directed to minimizing or partially or completely
inhibiting the development of the associated disease, pathological
condition, or disorder; and supportive treatment, that is,
treatment employed to supplement another specific therapy directed
toward the improvement of the associated disease, pathological
condition, or disorder. "Treating" also includes symptomatic
treatment, that is, treatment directed toward constitutional
symptoms of the associated disease, pathological condition, or
disorder. Thus "treating" includes submitting or subjecting a
subject to a compound which will promote the elimination or
reduction of a disease or symptoms of a disease, or which will slow
the progression of said disease. For example, a subject may be
treated with, synthesized organic molecules, naturally occurring
organic molecules, peptides, polypeptides, nucleic acid molecules,
and components thereof "Treating" also includes the act of not
giving a subject a contra-indicated therapeutic.
[0060] "Concurrent administration," "administration in
combination," "simultaneous administration" or "administered
simultaneously" as used herein, interchangeably mean that the
compounds are administered at the same point in time or immediately
following one another. In the latter case, the two compounds are
administered at times sufficiently close that the results observed
are indistinguishable from those achieved when the compounds are
administered at the same point in time.
[0061] "Enhance the activity of" when used herein to describe the
effect of a second active agent on a first active agent means the
second active agent is administered in an amount that will (a)
increase the treatment efficacy of an otherwise efficacious active
agent to an even more efficacious level, as well as (b) increase
the treatment efficacy of an otherwise ineffective active agent to
an efficacious level.
[0062] Tryptophan hydroxylase 2 (Tph2) is known and described at
GenBank accession numbers NM.sub.--173353 (human); NM.sub.--173391
(mouse); and NM.sub.--173839 (rat). See also M. Bader and D.
Walther, PCT Patent Application WO 2004/007704 (US
2006/0275759).
[0063] 1. Tph2 mutation or polymorphism detection/genotyping. In
general, the step of detecting the polymorphism of interest, or
genotyping a subject, may be carried out by collecting a biological
sample containing DNA from the subject, and then determining the
presence or absence of the polymorphism of interest in the
biological sample. Any biological sample which contains the DNA of
that subject may be employed, including tissue samples and blood
samples, with blood cells being a particularly convenient source.
The nucleotide sequence of the mouse and human Tph2 gene is known
and suitable probes, restriction enzyme digestion techniques, or
other means of detecting the polymorphism may be implemented based
on this known sequence, or the variations described herein, in
accordance with standard techniques. See, e.g., U.S. Pat. Nos.
6,027,896 and 5,767,248 to A. Roses et al.
[0064] In describing the mutations disclosed herein in the novel
proteins described herein, and the nucleotides encoding the same,
the naming method is as follows: [amino acid replaced] [amino acid
number in sequence of known protein][alternate amino acid]. For
example, for the mouse Tph2 variant disclosed herein, proline at
the 447th amino acid in the protein is replaced with arginine
(Zhang, X et al., 2004).
[0065] The polymorphisms described herein can be detected in
accordance with known techniques based upon the known sequence
information of the mouse and human Tph2 gene and the information
provided herein. Novel nucleic acid sequences and proteins
described herein can be isolated from human sources based upon the
information provided herein or produced by other means such as
site-directed mutagenesis of known or available amino acids,
coupled as necessary with techniques for the production of
recombinant proteins known in the art.
[0066] Determining the presence or absence of DNA containing a
polymorphism or mutation of interest may be carried out with an
oligonucleotide probe labeled with a suitable detectable group, or
by means of an amplification reaction such as a polymerase chain
reaction or ligase chain reaction (the product of which
amplification reaction may then be detected with a labeled
oligonucleotide probe or a number of other techniques). Further,
the detecting step may include the step of detecting whether the
subject is heterozygous or homozygous for the polymorphism of
interest. Numerous different oligonucleotide probe assay formats
are known which may be employed to carry out the present invention.
See, e.g., U.S. Pat. No. 4,302,204 to Wahl et al.; U.S. Pat. No.
4,358,535 to Falkow et al.; U.S. Pat. No. 4,563,419 to Ranki et
al.; and U.S. Pat. No. 4,994,373 to Stavrianopoulos et al.
(applicants specifically intend that the disclosures of all U.S.
Patent references cited herein be incorporated herein by
reference).
[0067] Amplification of a selected, or target, nucleic acid
sequence may be carried out by any suitable means. See generally D.
Kwoh and T. Kwoh, Am. Biotechnol. Lab. 8, 14-25 (1990). Examples of
suitable amplification techniques include, but are not limited to,
polymerase chain reaction, ligase chain reaction, strand
displacement amplification (see generally G. Walker et al., Proc.
Natl. Acad. Sci. USA 89, 392-396 (1992); G. Walker et al., Nucleic
Acids Res. 20, 1691-1696 (1992)), transcription-based amplification
(see D. Kwoh et al., Proc. Natl. Acad. Sci. USA 86, 1173-1177
(1989)), self-sustained sequence replication (or "3SR") (see J.
Guatelli et al., Proc. Natl. Acad. Sci. USA 87, 1874-1878 (1990)),
the Q.beta. replicase system (see P. Lizardi et al., BioTechnology
6, 1197-1202 (1988)), nucleic acid sequence-based amplification (or
"NASBA") (see R. Lewis, Genetic Engineering News 12 (9), 1 (1992)),
the repair chain reaction (or "RCR") (see R. Lewis, supra), and
boomerang DNA amplification (or "BDA") (see R. Lewis, supra).
[0068] DNA amplification techniques such as the foregoing can
involve the use of a probe, a pair of probes, or two pairs of
probes which specifically bind to DNA containing the polymorphism
of interest, but do not bind to DNA that does not contain the
polymorphism of interest under the same hybridization conditions,
and which serve as the primer or primers for the amplification of
the DNA or a portion thereof in the amplification reaction. Such
probes are sometimes referred to as amplification probes or primers
herein.
[0069] In general, an oligonucleotide probe which is used to detect
DNA containing a polymorphism or mutation of interest is an
oligonucleotide probe which binds to DNA encoding that mutation or
polymorphism, but does not bind to DNA that does not contain the
mutation or polymorphism under the same hybridization conditions.
The oligonucleotide probe is labeled with a suitable detectable
group, such as those set forth below in connection with antibodies.
Such probes are sometimes referred to as detection probes or
primers herein.
[0070] Probes and primers, including those for either amplification
and/or protection, are nucleotides (including naturally occurring
nucleotides such as DNA and synthetic and/or modified nucleotides)
are any suitable length, but are typically from 5, 6, or 8
nucleotides in length up to 40, 50 or 60 nucleotides in length, or
more. Such probes and or primers may be immobilized on or coupled
to a solid support such as a bead, chip, pin, or microtiter plate
well in accordance with known techniques, and/or coupled to or
labeled with a detectable group such as a fluorescent compound, a
chemiluminescent compound, a radioactive element, or an enzyme in
accordance with known techniques.
[0071] Polymerase chain reaction (PCR) may be carried out in
accordance with known techniques. See, e.g., U.S. Pat. Nos.
4,683,195; 4,683,202; 4,800,159; and 4,965,188. In general, PCR
involves, first, treating a nucleic acid sample (e.g., in the
presence of a heat stable DNA polymerase) with one oligonucleotide
primer for each strand of the specific sequence to be detected
under hybridizing conditions so that an extension product of each
primer is synthesized which is complementary to each nucleic acid
strand, with the primers sufficiently complementary to each strand
of the specific sequence to hybridize therewith so that the
extension product synthesized from each primer, when it is
separated from its complement, can serve as a template for
synthesis of the extension product of the other primer, and then
treating the sample under denaturing conditions to separate the
primer extension products from their templates if the sequence or
sequences to be detected are present. These steps are cyclically
repeated until the desired degree of amplification is obtained.
Detection of the amplified sequence may be carried out by adding to
the reaction product an oligonucleotide probe capable of
hybridizing to the reaction product (e.g., an oligonucleotide probe
of the present invention), the probe carrying a detectable label,
and then detecting the label in accordance with known techniques,
or by direct visualization on a gel. When PCR conditions allow for
amplification of all allelic types, the types can be distinguished
by hybridization with allelic specific probe, by restriction
endonuclease digestion, by electrophoresis on denaturing gradient
gels, or other techniques.
[0072] Ligase chain reaction (LCR) is also carried out in
accordance with known techniques. See, e.g., R. Weiss, Science 254,
1292 (1991). In general, the reaction is carried out with two pairs
of oligonucleotide probes: one pair binds to one strand of the
sequence to be detected; the other pair binds to the other strand
of the sequence to be detected. Each pair together completely
overlaps the strand to which it corresponds. The reaction is
carried out by, first, denaturing (e.g., separating) the strands of
the sequence to be detected, then reacting the strands with the two
pairs of oligonucleotide probes in the presence of a heat stable
ligase so that each pair of oligonucleotide probes is ligated
together, then separating the reaction product, and then cyclically
repeating the process until the sequence has been amplified to the
desired degree. Detection may then be carried out in like manner as
described above with respect to PCR.
[0073] It will be readily appreciated that the detecting steps
described herein may be carried out directly or indirectly. For
example, a polymorphism or mutation could be detected by measuring
by digestion with restriction enzymes, detection of markers that
are linked to the mutation or polymorphism, etc.
[0074] Genotype determinations can be compiled to predict either
prognosis, drug efficacy, or suitability of a patient for
participating in clinical trials of a neurological disease
therapeutic. For example, the genotype may be compiled with other
patient parameters such as age, sex, disease diagnosis, and known
allelic frequency of a representative control population. A
determination of the statistical probability of the patient having
a particular disease risk, drug response, or patient outcome may be
assessed from such genotype determinations. Patient outcome, i.e. a
prediction of a patient's likely health status, may include a
prediction of the patient's response to therapy, rehabilitation
time, recovery time, cure rate, rate of disease progression,
predisposition for future disease, or risk of having relapse.
[0075] Kits useful for carrying out the methods of the present
invention will, in general, comprise one or more oligonucleotide
probes and other reagents for carrying out the methods as described
above, such as restriction enzymes, optionally packaged with
suitable instructions for carrying out the methods.
[0076] Particular examples of Tph2 mutations that can be screen for
or determined in carrying out the present invention include but are
not limited to mutations that encode a change in an amino acid of
the encoded protein, said amino acid selected from the group
consisting of A65, V66, F68, L77, F84, 194, R97, E105, P152, W153,
P155, D162, L175, R191, E211, V223, P244, G251, R276, P277, V278,
R285, R294, P308, Y310, E313, A333, 1339, A342, S343, L344, A346,
K353, V421, E423, A428, A436, R441, Y446, P449, Y450, and Q468. A
mutation inducing any change in the normal sequence (the amino acid
to the left of the number identifying the amino acid location) is
within the scope of the present invention (for example, a change
of: I to T; E to G, P to Q; R to Q; G to V; P to L; R to W; Y to D;
E to K; L to P; R to Q; Y to C; R to H; P to R; A to V; V to I; L
to V; Q to X (where X is a stop codon) etc.).
[0077] Particularly preferred are the human P449R mutation, the
human R441H mutation the human W153R mutation, the human A65V
mutation, the human V661 mutation, the human L175V mutation, and
the human Q468X (where X is a stop codon) mutation. Subjects may be
determined to be heterozygous or homozygous for the indicated
mutation, or may be determined to carry different mutations on the
same, or different, alleles.
[0078] Note that 43 mutations are identified herein, some
identified by sequence analysis and some by sequence identity
compared to PAH. Table 1 below lists corresponding mutations in
Tph2 in six different species, which corresponding mutations are
also useful for carrying out the present invention. It is striking
that all 43 amino acids for these mutations are virtually identical
in Tph2 in six different species (except in two positions). This
strongly indicates that mutations in similar position in Tph2 have
a like functional impact on serotonin production in other
species.
TABLE-US-00001 TABLE 1 Corresponding mutations in additional
species. human mouse rat chicken zebrafish pufferfish A65 A63 A60
A64 A35 A61 V66 V64 V61 V65 V36 V62 F68 F66 F63 F67 F38 F64 L77 L75
L72 L76 L47 L73 F84 F82 F79 F83 F54 F80 I94 I92 I89 I93 I64 I90 R97
R95 R92 R96 R67 R93 E105 E103 E100 E104 E75 E101 P152 P150 P147
P151 P135 P148 W153 W151 W148 W152 W136 W149 P155 P153 P150 P154
P138 P151 D162 D160 D157 D161 D145 D158 L175 L173 L170 L174 L158
L171 R191 R189 R186 R190 R174 R187 E211 E209 E206 E210 E194 E207
V223 V221 V218 V222 V206 V219 P244 P242 P239 P243 P227 P240 G251
G249 G246 G250 G234 G247 R276 R274 R271 R275 R259 R272 P277 P275
P272 P276 P260 P273 V278 V276 V273 V277 V261 V274 R285 R283 R280
R284 R268 R281 R294 R292 R289 R293 R277 R290 P308 P306 P303 P307
P291 P304 Y310 Y308 Y305 Y309 Y293 Y306 E313 E311 E308 E312 Y296
E309 A333 A331 A328 A332 A316 A329 I339 I337 I334 I338 I322 I335
A342 A340 A337 A341 A325 A338 S343 S341 S338 S342 S326 S339 L344
L342 L339 L343 L327 L340 A346 A344 A341 A345 A329 A342 K353 K351
K348 K352 K336 K349 V421 V419 V416 V420 V404 V417 E423 E418 E422
E406 E419 A428 A426 A423 A427 A411 A424 A436 A434 A431 A435 A419
A432 R441 R439 R436 R440 R424 R437 Y446 Y444 Y441 Y445 Y429 Y442
P449 P447 P444 P448 P432 P445 Y450 Y448 Y445 Y449 Y433 Y446 Q468
Q466 Q463 Q467 Q464
[0079] In addition to the foregoing, intronic mutations are useful
for carrying out the present invention. For example, a G->A
mutation at position 144 in Intron 6 of human TPH2 is useful for
carrying out the present invention. This is a non-coding mutation
in the Intron 6 and is predicted to cause an alternative splicing
variant/mutation of human TPH2. The entire intron 6 in human TPH2
is 6236 base pairs in length; the pertinent portion of exon 6 is
shown in panel a of Table 2 below, and the pertinent portion of the
mutant intron 6 is shown in panel B of Table 2 below.
TABLE-US-00002 TABLE 2 Intron 6 non-coding mutation. A
GGTCAGCCCATTCCCAGGGTGGAGTATACTGAAGA (SEQ ID NO:1)
AGAAACTAAAACTTGGGGTGTTGTATTCCGGGAGC
TCTCCAAACTCTATCCCACTCATGCTTGCCGAGAG
TATTTGAAAAACTTCCCTCTGCTGACTAAATACTG
TGGCTACAGAGAGGACAATGTGCCTCAACTCGAAG ATGTCTCCATGTTTCTGAAAG B
gtaagatttcacacaggctgtctcttattagtcaa (SEQ ID NO:2)
tatcctcaattgccttccaaggacacaggttgcag
caatggctctttttccaaaaaaggaaaaacagtga
tttaaaaaattgttggctttgagccaacaattacc
tgcggccacctgtgggaagcagagcaagggactca
gctgcttttgcagctcaggagcttgctgaggcctc
tttgtggctggttgttgtaaatggtaaggcccaaa
ggatatttgcaagttcagctctgagctttttctga
tccaggagctgctgtgctgggctacatgagtatga
aatgacctccaaaagtgcctttttatttgctttgt taaaaagtat The g/a polymorphism
is shown in bold underlined font in panel b.
[0080] 2. Active agents. Active agents used to carry out the
present invention are, in general, serotonin enhancers. Numerous
serotonin enhancers and serotonin enhancer therapies are known.
See, e.g., U.S. Pat. No. 6,218,395.
[0081] The serotonin enhancer can be a serotonin reuptake inhibitor
or selective serotonin reuptake inhibitor, such as described in
U.S. Pat. Nos. 6,552,014; 6,492,366; 6,387,956; 6,369,051; or
5,958,429. Examples of known serotonin reuptake inhibitors that may
be used in carrying out the present invention include, but are not
limited to: [0082] cianopramine or a pharmaceutically acceptable
salt thereof (e.g.,
5-[3-(dimethylamino)propyl]-10.11-dihydro-5H-dibenz[b,f]azepine-3-carboni-
trile); [0083] citalopram or a pharmaceutically acceptable salt
thereof (e.g.,
1-[3-(dimethylamino)propyl]-1-(p-fluorophenyl)-1,3-dihydro-isobenz-
ofuran-5-carbonitrile); [0084] escitalopram or a pharmaceutically
acceptable salt thereof (e.g.,
(S)-1-3-dimethylamino-propyl-1-(4'-fluoro-phenyl)-1,3-dihydro-isobenzofur-
an-5-carbonitrile,oxalate); [0085] dapiprazole or a
pharmaceutically acceptable salt thereof (e.g., 5, 6, 7,
8-tetrahydro-3-[2-(4-o-tolyl-1-piperazinyl)ethyl]-1,2,4-triazolo[4,3-a]py-
ridine (hydrochloride)); [0086] desvenlafaxine or a
pharmaceutically acceptable salt thereof (e.g., Phenol,
4-[2-(dimethylamino)-1-(1-hydroxycyclohexyl)ethyl]-(Z)-2-butanedioate
(1:1) monohydrate); [0087] duloxetine or a pharmaceutically
acceptable salt thereof, including LY 223.332, LY264.452
((-)-enantiomer), LY248.686 (HCl), and Y227.942 (+)-enantiomer)
(e.g., (+)-(S)--N-methyl-1-(1-naphtyloxy)-2-thiophenepropylamine
(HCl)); [0088] fluoxetine or a pharmaceutically acceptable salt
thereof (e.g.,
3-[(p-trifluoromethyl)phenoxy]-N-methyl-3-phenyl-propylamine(hydrochlorid-
e)); [0089] fluvoxamine or a pharmaceutically acceptable salt
thereof (e.g., (E)-5-methoxy-4'-(trifluoromethyl)valerophenone
O-(2-amino-ethyl)oxime (hydrogen maleate)); [0090] ifoxetine or a
pharmaceutically acceptable salt thereof (e.g.,
(+/-)-cis-4-(2,3-xylyloxy)-3-piperidinol (sulfate)); [0091]
indalpine or a pharmaceutically acceptable salt thereof (e.g.,
3-[2-(4-piperidyl)ethyl]indole); [0092] LY 113.821 or a
pharmaceutically acceptable salt thereof (e.g.,
N-methyl-3-(1-naphthoxy)-3-phenylpropylamine); [0093] mirtazapine
or a pharmaceutically acceptable salt thereof (e.g., 1, 2, 3, 4,
10.14-hexahydro-2 methylpiprazino[2,1-a]pyrido[2,3-c]benzazepine);
[0094] nefazodone or a pharmaceutically acceptable salt thereof
(e.g.,
1-[3-[4-(m-chlorophenyl)-1-piperazinyl]propyl]-3-ethyl-4-(2-phenoxyethyl)-
-.sup.2-1,2,4-triazolin-5-one (HCl)); [0095] 2-nitroimipramine or a
pharmaceutically acceptable salt thereof (e.g.,
5-[3-(dimethylamino)propyl]-2-nitro-10.11-dihydro-5H-dibenz[b,f]azepine
(hydrochloride); [0096] nortriptyline or a pharmaceutically
acceptable salt thereof (e.g.,
10.11-dihydro-N-methyl-5H-dibenzo[a,d]cycloheptene-.DELTA..sup.5,.GAMMA.p-
ropylamine (hydrochloride)); [0097] paroxetine or a
pharmaceutically acceptable salt thereof (e.g.,
(3S-trans)-3-[(1,3-benzodioxol-5-yloxy)
methyl]-4-(4-fluorophenyl)piperidine)); [0098] RU 25.591 or a
pharmaceutically acceptable salt thereof (e.g.,
cis-6,7,8,9-tetrahydro-N,N-dimethyl-5-(p-nitrophenoxy)-5H-benzocyclohepte-
n-7-amine(fumarate)); [0099] sercloremine or a pharmaceutically
acceptable salt thereof (e.g.,
4-(5-chloro-2-benzofuranyl)-1-methylpiperidine (hydrochloride));
[0100] sertraline or a pharmaceutically acceptable salt thereof
(e.g.,
(+)-cis(1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydro-N-methyl-1-napht-
hylamine(HCl)); [0101] setiptiline or a pharmaceutically acceptable
salt thereof (e.g.,
2,3,4,9-tetrahydro-2-methyl-1H-dibenzo[c,f]cyclohepta[1,2-c]pyridine
(maleate)); [0102] tianeptine or a pharmaceutically acceptable salt
thereof (e.g.,
N-(3-chloro-6,11-dihydro-6-methyldibenzo[c,f][1,2]thiazepin-11-yl)-7-amin-
o-heptanoic acid S,S-dioxide); [0103] trazodone or a
pharmaceutically acceptable salt thereof (e.g.,
2-[3-[4-(3-chlorophenyl)-1-piperazinyl]propyl]-1,2,4-triazolo[4,3-a]pyrid-
in-3(2H)-one (hydrochloride)); [0104] venlafaxine or a
pharmaceutically acceptable salt thereof (e.g.,
(+/-)-1-[2-(dimethylamino)-1-(p-methoxyphenyl)ethyl]cyclohexan-1-ol
(HCl)) [0105] viqualine or a pharmaceutically acceptable salt
thereof (e.g.,
cis-6-methoxy-4-[3-(3R,4R)-(3-vinylpiperidyl)propyl]-quinoline);
and [0106] zimeldine or a pharmaceutically acceptable salt thereof
(e.g.,
(Z)-3-(p-bromophenyl)-N,N-dimethyl-3-(3-pyridyl)allylamine).
[0107] The serotonin enhancer can be a monoamine oxidase inhibitor
such as described in U.S. Pat. Nos. 6,472,423 and 6,011,054.
Examples of monoamine oxidase inhibitors include but are not
limited to Isocarboxazid, phenelzine, and tranylcypromine. The
serotonin enhancer can be a serotonin agonist such as described in
U.S. Pat. Nos. 6,656,172; 6,579,899 and 6,387,907. The serotonin
enhancer can be an amphetamine (including derivatives thereof such
as phentermine, fenfluramine, and
(+)-3,4-methylenedioxyamphetamine. The serotonin enhancer can be a
tricyclic antidepressant such as described in U.S. Pat. Nos.
6,368,814; 6,358,944; 6,239,162; and 6,211,171. Examples of
tricyclic antidepressants include but are not limited to
imipramine, amitriptyline and clomipramine.
[0108] The serotonin enhancer can be an anxiolytic such as
buspirone or ipsapirone.
[0109] The serotonin enhancer can be a precursor or prodrug of
serotonin, or an intermediate in serotonin biosynthesis, such as
described in U.S. Pat. Nos. 6,579,899; 6,013,622; and 5,595,772.
Examples includes tryptophan, 5-hydroxytryptophan, TPH2 co-factor
tetrahydrobiopterin and its precursors, a tryptophan-rich diet or
dietary supplements of tryptophan.
[0110] Decarboxylase inhibitors, particularly peripheral
decarboxylase inhibitors, useful for carrying out the present
invention include carbidopa (particularly L-carbidopa) and/or
benserazide. See, e.g., U.S. Pat. No. 6,387,936.
[0111] Tetrahydrobiopterin
[(6R)-L-erythro-5,6,7,8-tetrahydrobiopterin (or "BH4"), useful as
an active agent in the some embodiments of the present invention,
is known. See, e.g., U.S. Pat. Nos. 6,451,788 and 4,920,122. While
the present invention is described primarily with reference to BH4,
other BH4 enhancers such as folates and analogs thereof, (e.g.,
methylfolate) as well as estrogen agonists and glucocorticoid
antagonists may be used in addition, or in alternative, thereto.
Folate analogs are known and described in, for example, U.S. Pat.
Nos. 6,808,725; 6,673,381; 6,500,829; and 6,191,133.
3. PHARMACEUTICAL FORMULATIONS
[0112] The active compounds described above may be formulated alone
or in combination with each other (e.g., in the combinations
described above) for administration in a pharmaceutical carrier in
accordance with known techniques. See, e.g., Remington, The Science
And Practice of Pharmacy (9.sup.th Ed. 1995).
[0113] In the manufacture of a pharmaceutical formulation according
to the invention, the active compound (including the
physiologically acceptable salts thereof) is typically admixed
with, inter alia, an acceptable carrier. The carrier must, of
course, be acceptable in the sense of being compatible with any
other ingredients in the formulation and must not be deleterious to
the patient. The carrier may be a solid or a liquid, or both, and
is preferably formulated with the compound as a unit-dose
formulation, for example, a tablet, which may contain from 0.01 or
0.5% to 95% or 99% by weight of the active compound or compounds.
One or more active compounds may be incorporated in the
formulations of the invention, which may be prepared by any of the
well known techniques of pharmacy comprising admixing the
components, optionally including one or more accessory
ingredients.
[0114] The formulations of the invention include those suitable for
oral, rectal, topical, buccal (e.g., sub-lingual), parenteral
(e.g., subcutaneous, intramuscular, intradermal, or intravenous),
topical (i.e., both skin and mucosal surfaces, including airway
surfaces) and transdermal administration, although the most
suitable route in any given case will depend on the nature and
severity of the condition being treated and on the nature of the
particular active compound which is being used.
[0115] Formulations suitable for oral administration may be
presented in discrete units, such as capsules, cachets, lozenges,
or tablets, each containing a predetermined amount of the active
compound; as a powder or granules; as a solution or a suspension in
an aqueous or non-aqueous liquid; or as an oil-in-water or
water-in-oil emulsion. Such formulations may be prepared by any
suitable method of pharmacy which includes the step of bringing
into association the active compound and a suitable carrier (which
may contain one or more accessory ingredients as noted above). In
general, the formulations of the invention are prepared by
uniformly and intimately admixing the active compound with a liquid
or finely divided solid carrier, or both, and then, if necessary,
shaping the resulting mixture. For example, a tablet may be
prepared by compressing or molding a powder or granules containing
the active compound, optionally with one or more accessory
ingredients. Compressed tablets may be prepared by compressing, in
a suitable machine, the compound in a free-flowing form, such as a
powder or granules optionally mixed with a binder, lubricant, inert
diluent, and/or surface active/dispersing agent(s). Molded tablets
may be made by molding, in a suitable machine, the powdered
compound moistened with an inert liquid binder.
[0116] Formulations suitable for buccal (sub-lingual)
administration include lozenges comprising the active compound in a
flavoured base, usually sucrose and acacia or tragacanth; and
pastilles comprising the compound in an inert base such as gelatin
and glycerin or sucrose and acacia.
[0117] Formulations of the present invention suitable for
parenteral administration comprise sterile aqueous and non-aqueous
injection solutions of the active compound, which preparations are
preferably isotonic with the blood of the intended recipient. These
preparations may contain anti-oxidants, buffers, bacteriostats and
solutes which render the formulation isotonic with the blood of the
intended recipient. Aqueous and non-aqueous sterile suspensions may
include suspending agents and thickening agents. The formulations
may be presented in unit\dose or multi-dose containers, for example
sealed ampoules and vials, and may be stored in a freeze-dried
(lyophilized) condition requiring only the addition of the sterile
liquid carrier, for example, saline or water-for-injection
immediately prior to use. Extemporaneous injection solutions and
suspensions may be prepared from sterile powders, granules and
tablets of the kind previously described. For example, in one
aspect of the present invention, there is provided an injectable,
stable, sterile composition comprising a active agent, in a unit
dosage form, in a sealed container. The compound or salt is
provided in the form of a lyophilizate which is capable of being
reconstituted with a suitable pharmaceutically acceptable carrier
to form a liquid composition suitable for injection thereof into a
subject. The unit dosage form typically comprises from about 10 mg
to about 10 grams of the compound or salt. When the compound or
salt is substantially water-insoluble, a sufficient amount of
emulsifying agent which is physiologically acceptable may be
employed in sufficient quantity to emulsify the compound or salt in
an aqueous carrier. One such useful emulsifying agent is
phosphatidyl choline.
[0118] Formulations suitable for rectal administration are
preferably presented as unit dose suppositories. These may be
prepared by admixing the active compound with one or more
conventional solid carriers, for example, cocoa butter, and then
shaping the resulting mixture.
[0119] Formulations suitable for topical application to the skin
preferably take the form of an ointment, cream, lotion, paste, gel,
spray, aerosol, or oil. Carriers which may be used include
petroleum jelly, lanoline, polyethylene glycols, alcohols,
transdermal enhancers, and combinations of two or more thereof.
[0120] For aerosol delivery to the airway surfaces of a patient the
pharmaceutical (e.g., as a dry powder or as liquid particles) the
formulation may be prepared in aerosolizable form and provided in
an aerosol or inhalation delivery device.
[0121] Formulations suitable for transdermal administration may be
presented as discrete patches adapted to remain in intimate contact
with the epidermis of the recipient for a prolonged period of time.
Formulations suitable for transdermal administration may also be
delivered by iontophoresis (see, for example, Pharmaceutical
Research 3 (6):318 (1986)) and typically take the form of an
optionally buffered aqueous solution of the active compound.
Suitable formulations comprise citrate or bis\tris buffer (pH 6) or
ethanol/water and contain from 0.1 to 0.2M active ingredient.
[0122] Further, the present invention provides liposomal
formulations of the compounds disclosed herein and salts thereof.
The technology for forming liposomal suspensions is well known in
the art. When the compound or salt thereof is an aqueous-soluble
salt, using conventional liposome technology, the same may be
incorporated into lipid vesicles. In such an instance, due to the
water solubility of the compound or salt, the compound or salt will
be substantially entrained within the hydrophilic center or core of
the liposomes. The lipid layer employed may be of any conventional
composition and may either contain cholesterol or may be
cholesterol-free. When the compound or salt of interest is
water-insoluble, again employing conventional liposome formation
technology, the salt may be substantially entrained within the
hydrophobic lipid bilayer which forms the structure of the
liposome. In either instance, the liposomes which are produced may
be reduced in size, as through the use of standard sonication and
homogenization techniques.
[0123] Of course, the liposomal formulations containing the
compounds disclosed herein or salts thereof, may be lyophilized to
produce a lyophilizate which may be reconstituted with a
pharmaceutically acceptable carrier, such as water, to regenerate a
liposomal suspension.
[0124] Other pharmaceutical compositions may be prepared from the
water-insoluble compounds disclosed herein, or salts thereof, such
as aqueous base emulsions. In such an instance, the composition
will contain a sufficient amount of pharmaceutically acceptable
emulsifying agent to emulsify the desired amount of the compound or
salt thereof. Particularly useful emulsifying agents include
phosphatidyl cholines, and lecithin.
[0125] In addition to active compounds or their salts, the
pharmaceutical compositions may contain other additives, such as
pH-adjusting additives. In particular, useful pH-adjusting agents
include acids, such as hydrochloric acid, bases or buffers, such as
sodium lactate, sodium acetate, sodium phosphate, sodium citrate,
sodium borate, or sodium gluconate. Further, the compositions may
contain microbial preservatives. Useful microbial preservatives
include methylparaben, propylparaben, and benzyl alcohol. The
microbial preservative is typically employed when the formulation
is placed in a vial designed for multidose use. Of course, as
indicated, the pharmaceutical compositions of the present invention
may be lyophilized using techniques well known in the art.
[0126] The compositions of this invention can also be administered
as a food based preparation, functional food, dietary supplement or
nutraceutical. For the purposes of this application, "functional
food" is defined as a food engineered or supplemented to give
improved nutritional value, "dietary supplement" is defined as a
substance produced by isolation, or microbial culture purification
that gives health benefits, and "nutraceutical" is defined as a
food, or parts of a food, that provide medical or health benefits,
including prevention and treatment of clinical conditions and/or
symptoms related thereto.
[0127] Active agents may be included in a pharmaceutical
formulation in an amount ranging from 0.001, 0.01, 0.1 or 1 percent
by weight, up to about 10, 20, 50 percent by weight, or more,
depending upon the activity of the particular active agent. For
example, in some embodiments a pharmaceutical formulation of the
invention comprises, consists of, or consists essentially of:
[0128] (a) from 0.001, 0.01 or 0.1 to 20 or 40 percent by weight of
a serotonin enhancer such as a serotonin reuptake inhibitor; [0129]
(b) from 0.1 or 1 to 40 or 60 percent by weight of a serotonin
precursor such as 5-hydroxytryptophan; [0130] (c) optionally,
0.001, 0.01 or 0.1 to 20 or 40 percent by weight of a peripheral
decarboxylase inhibitor such as carbidopa; [0131] (d) optionally,
from 0.1 or 1 to 40 or 60 percent by weight of a BH4 enhancer such
as tetrahydrobiopterin; and [0132] (e) from 1 to 99.9 percent by
weight of a pharmaceutically acceptable carrier. In some other
exemplary embodiments, a pharmaceutical formulation of the
invention comprises, consists of, or consists essentially of:
[0133] (a) from 0.001, 0.01 or 0.1, up to 20 or 40 percent by
weight of a serotonin enhancer such as a serotonin reuptake
inhibitor; [0134] (b) from 0.001, 0.01 or 0.1, up to 20 or 40
percent by weight of decarboxylase inhibitor such as carbidopa;
[0135] (c) optionally, from 0.1 or 1 to 40 or 60 percent by weight
of a BH4 enhancer such as tetrahydrobiopterin; [0136] (d)
optionally, from 0.1 or 1 to 40 or 60 percent by weight of a
serotonin precursor such as 5-hydroxytryptophan; and [0137] (e)
from 1 to 99.9 percent by weight of a pharmaceutically acceptable
carrier. In some other exemplary embodiments a pharmaceutical
formulation of the invention comprises, consists of, or consists
essentially of [0138] (a) 0.001, 0.01 or 0.1 to 20 or 40 percent by
weight of a serotonin enhancer such as a serotonin reuptake
inhibitor; [0139] (b) from 0.1 or 1 to 40 or 60 percent by weight
of a BH4 enhancer such as tetrahydrobiopterin; [0140] (c)
optionally, 0.001, 0.01 or 0.1 to 20 or 40 percent by weight of a
peripheral decarboxylase inhibitor such as carbidopa; [0141] (d)
optionally, from 0.1 or 1 to 40 or 60 percent by weight of a
serotonin precursor such as 5-hydroxytryptophan; and [0142] (e)
from 1 to 99.9 percent by weight of a pharmaceutically acceptable
carrier. In other exemplary embodiments, a pharmaceutical
formulation of the invention comprises, consists of, or consists
consisting essentially of: [0143] (a) from 0.1 or 1 to 40 or 60
percent by weight of a serotonin precursor such as
5-hydroxytryptophan; and [0144] (b) from 0.001, 0.01 or 0.1, up to
20 or 40 percent by weight of decarboxylase inhibitor such as
carbidopa; [0145] (c) optionally, from 0.1 or 1 to 40 or 60 percent
by weight of a BH4 enhancer such as tetrehydrobiopterin; [0146] (d)
from 1 to 99.9 percent by weight of a pharmaceutically acceptable
carrier. And in still other embodiments, a pharmaceutical (this
term including neutraceutical) formulation of the invention
comprises, consists of, or consists essentially of: [0147] (a) from
0.1 or 1 to 40 or 60 percent by weight of a serotonin precursor
such as 5-hydroxytryptophan; and [0148] (b) from 0.1 or 1 to 40 or
60 percent by weight of a BH4 enhancer such as tetrahydrobiopterin;
and [0149] (c) from 1 to 99.9 percent by weight of a
pharmaceutically acceptable carrier (in the case of a
neutraceutical optionally from 1 to 99.9 percent by weight of other
food ingredients such as fats, proteins, carbohydrates, and
combinations thereof).
[0150] Such formulations may be provided in any suitable form
including those described above, such as tablets, capsules,
suppositories, inhalation or aerosolizable formulations,
formulations in an inhalation delivery device, parenterally
injectable formulations. etc. In the case of a neutraceutical
composition, the formulation may be provided in the form of a bar,
beverage, drink, snack food, etc.
4. SUBJECTS, DOSAGE AND ROUTES OF ADMINISTRATION
[0151] Subjects to be treated by the methods of the invention are,
in some embodiments (such as those involving administration of a
serotonin enhancer such as a serotonin reuptake inhibitor in
combination with a serotonin precursor and/or a peripheral
decarboxylase inhibitor) those exhibiting symptoms of a serotonin
neurotransmission dysregulation disorder (particularly depression,
anxiety, or substance abuse, although it will be understood that
subjects afflicted with other serotonin neurotransmission
dysregulation disorders such as set forth above may be treated as
well). Subjects may be those previously determined to be
non-responsive or insufficiently responsive to treatment therapy
with a serotonin enhancer such as a serotonin reuptake inhibitor
alone (e.g., subjects who exhibited no benefit or improvement in
symptoms to administration of a serotonin enhancer such as a
serotonin reuptake inhibitor, or subjects who exhibited
insufficient benefit or improvement in symptoms to administration
of a serotonin enhancer such as a serotonin reuptake
inhibitor).
[0152] Subjects may, in some preferred embodiments, be genotyped or
determined to carry a Tph2 mutation prior to initiation of the
treatments described herein. Such subjects may be more confidently
administered the treatments described herein based on a greater
expectation of likely clinical benefit of these treatments, based
on the greater understanding of the underlying genetic and
physiological basis of the disorder as described herein.
[0153] The active compounds may be administered to the subject by
any suitable route, including those indicated with respect to
particular formulations above. Thus suitable routes of
administration include oral administration, buccal administration,
parenteral injection, inhalation or aerosol administration,
transcutaneous administration, etc.
[0154] The therapeutically effective dosage of any specific active
compound, the use of which is in the scope of present invention,
will vary somewhat from compound to compound, and patient to
patient, and will depend upon the condition of the patient and the
route of delivery.
[0155] For example, a serotonin enhancer such as a serotonin
reuptake inhibitor can be administered to the average adult human
for the treatment of the disorders described herein in an amount of
from about 0.1 mg to about 2000 mg, preferably from about 1 mg to
about 200 mg per unit dose.
[0156] A peripheral decarboxylase inhibitor such as carbidopa or
benserazide can, in some embodiments, be administered to an average
adult human subject in an amount of from 5 or 10 to 25, 50 or 100
milligrams per unit dose.
[0157] A precursor or prodrug of serotonin, or an intermediate in
serotonin biosynthesis, such as 5-hydroxytryptophan, may be
administered in an amount of from about 1, 2 5 or 10 milligrams up
to 0.1, 0.5, 1 or 4 grams per unit dose.
[0158] Administration of unit doses of active agents, alone or in
combination, may be one or several times daily, for example 2, 3, 4
or 8 times, giving for example, 1, 2 or 3 doses each time.
[0159] The present invention is explained in greater detail in the
following non-limiting Examples.
EXAMPLES
[0160] TPH1 and 2 convert tryptophan to 5-hydroxytryptophan, which
is then decarboxylated by aromatic amino acid decarboxylase (AADC)
to serotonin. The two enzymes belong to the family of aromatic
amino acid hydroxylases. Members of this family include
phenylalanine hydroxylase (PAH) and tyrosine hydroxylase (TH)
respectively involved in the synthesis of tyrosine and L-Dopa, the
precursor of dopamine. Both TH and TPH catalyze the rate limiting
steps in the synthesis of dopamine and serotonin. There is a high
degree of sequence conservation in the genes for these four
different enzymes. These enzymes show considerable structural
similarities and require the same co-factor for function
(Fitzpatrick, 1999). Over 400 mutations have been previously
identified in the PAH gene and these mutations, which result in
reduced enzyme activity, produce hyperphenylalaninemia causing of
one of the most prevalent condition identified in newborns,
phenylketonuria (PKU) (Scriver et al, 2003).
[0161] A (G1463A) SNP which replaces a highly conserved Arg441
(CGT) with His441 (CAT) has been identified resulting in .about.80%
decrease in serotonin levels as compared to wild type TPH2 when
expressed in PC12 cells (FIG. 1), indicating a severe
loss-of-function phenotype of TPH2 (R441H) in serotonin synthesis.
In addition, allele-specific genotyping has identified that 9
subjects were found to carry the 1463A allele in a cohort of 87
subjects diagnosed with unipolar major depression, while only 3
subjects from a cohort of non-unipolar major depression (or
control) subjects (N=279) were found to carry the 1463A allele
(Table 3). The presence of this functional SNP in unipolar major
depression subjects versus non-unipolar major depression subjects
was statistically significant (P=1.989E-04 by Fisher's Exact Test).
Moreover, among these 9 unipolar major depression subjects, 7 had
family history of mental illness or drug and alcohol abuse, 6 bad
suicidal ideation or attempt and 4 had generalized anxiety
symptoms. Interestingly, 7 subjects exhibited lack of
responsiveness to SSRI, while 2 subjects were responsive only to
high doses of SSRI (Table 3). However, among the 3 non-unipolar
major depression (or control) subjects carrying 1463A allele, one
subject had generalized anxiety symptoms, while the other two had
mild depression and family history of mental illness or drug and
alcohol abuse (Table 3), indicating a higher susceptibility for
certain neuropsychiatric disorders in the presence of 1463A allele.
Therefore, identification of a loss-of-function mutation in hTPH2
provides direct evidence for a role of brain serotonin synthesis in
unipolar major depression.
TABLE-US-00003 TABLE 3 Summary of genotype and history of subjects.
Patient Family SSRI I.D. Sex Age Allele History Suicidality Anxiety
Response Note Unipolar Major Depression Patients N = 87 1202 F 72
A/A - + - + Sertraline 200 mg 1294 M 80 G/A + + - - 1496 M 74 A/A -
- + + Sertraline 100 mg 1745 M 71 G/A + - + - 1747 M 82 G/A + + - -
1839 F 69 G/A + - - - 1851 F 65 G/A + + - - 1902 F 77 A/A + + + -
1975 M 64 G/A + + + - Non-Unipolar Major Depression Subjects N =
279 1174 A/A - + 1541 G/A + - Mild depression 1996 G/A + - Mild
depression Family history: family history for mental illness, drug
or alcohol abuse Suicidality: suicidal ideation or attempt Anxiety:
three or more anxiety symptoms
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[0175] The foregoing is illustrative of the present invention, and
is not to be construed as limiting thereof. The invention is
defined by the following claims, with equivalents of the claims to
be included therein.
Sequence CWU 1
1
21196DNAHomo sapiens 1ggtcagccca ttcccagggt ggagtatact gaagaagaaa
ctaaaacttg gggtgttgta 60ttccgggagc tctccaaact ctatcccact catgcttgcc
gagagtattt gaaaaacttc 120cctctgctga ctaaatactg tggctacaga
gaggacaatg tgcctcaact cgaagatgtc 180tccatgtttc tgaaag
1962360DNAHomo sapiens 2gtaagatttc acacaggctg tctcttatta gtcaatatcc
tcaattgcct tccaaggaca 60caggttgcag caatggctct ttttccaaaa aaggaaaaac
agtgatttaa aaaattgttg 120gctttgagcc aacaattacc tgcggccacc
tgtgggaagc agagcaaggg actcagctgc 180ttttgcagct caggagcttg
ctgaggcctc tttgtggctg gttgttgtaa atggtaaggc 240ccaaaggata
tttgcaagtt cagctctgag ctttttctga tccaggagct gctgtgctgg
300gctacatgag tatgaaatga cctccaaaag tgccttttta tttgctttgt
taaaaagtat 360
* * * * *