METHOD OF TREATMENT OF HIV OR AIDS
The present invention relates to methods of treatment
of HIV or AIDS.
Background
Human immunodeficiency virus (HIV) is a lentivirus (a
member of the retrovirus family) that causes acguired
immunodeficiency syndrome (AIDS) , a condition in humans in
which the immune system begins to fail, leading to lifethreatening
opportunistic infections.
HIV infects primarily vital cells in the human immune
system such as CD4 + T cells) , macrophages and dendritic
cells. HIV infection leads to low levels of CD4 + T cells
through three main mechanisms: First, direct viral killing
of infected cells; second, increased rates of apoptosis in
infected cells; and third, killing of infected CD4 + T cells
by CD8 cytotoxic lymphocytes that recognize infected cells.
When CD4 + T cell numbers decline below a critical level,
cell mediated immunity is lost, and the body becomes
progressively more susceptible to opportunistic infections.
Most untreated people infected with HIV-1 eventually
develop AIDS. These individuals mostly die from
opportunistic infections or malignancies associated with the
progressive failure of the immune system. HIV progresses to
AIDS at a variable rate affected by viral, host, and
environmental factors; most will progress to AIDS within 10
years of HIV infection: some will have progressed much
sooner, and some will take much longer. Treatment with antiretrovirals
increases the life expectancy of people infected
with HIV. Even after HIV has progressed to diagnosable AIDS,
the average survival time with antiretroviral therapy was
estimated to be more than 5 years as of 2005. Without
antiretroviral therapy, someone who has AIDS typically dies
within a year.
Current treatment for HIV infection consists of highly
active antiretroviral therapy, or HAART . This has been
highly beneficial to many HIV-infected individuals since its
introduction in 1996, when the protease inhibitor-based
HAART initially became available. Current HAART options are
combinations (or "cocktails") consisting of at least three
drugs belonging to at least two types, or "classes, " of
antiretroviral agents. Typically, these classes are two
nucleoside analogue reverse transcriptase inhibitors (NARTIs
or NRTIs) plus either a protease inhibitor or a nonnucleoside
reverse transcriptase inhibitor (NNRTI) .
Despite the success of HAART in controlling HIV
infection and reducing HIV-associated mortality, current
drug regimens are unable to completely eradicate HIV
infection. Many people on HAART achieve suppression of HIV
to levels below the limit of detection of standard clinical
assays for many years. However, upon withdrawal of HAART,
HIV viral loads rebound quickly with a concomitant decline
in CD4+ T cells, which, in most cases, absent a resumption
of treatment, leads to AIDS and death.
It is an aim of an embodiment of the invention to
provide a treatment for HIV or AIDs.
Summary
A first aspect of the invention provides a method of
treating HIV or AIDS comprising administering a composition
comprising a polypeptide comprising LKKTETQ (SEQ ID NO: 1 )
or a variant thereof.
An alternative first aspect provides a composition
comprising a polypeptide comprising LKKTETQ (SEQ ID NO: 1 )
or a variant thereof for treating HIV or AIDS.
An alternative first aspect provides use of a
composition comprising a polypeptide comprising LKKTETQ (SEQ
ID NO: 1 ) or a variant thereof in the manufacture of a
medicament for treating HIV or AIDS.
A second aspect of the invention provides a method of
treating HIV or AIDS comprising administering a composition
comprising a peptidomimetic of a polypeptide comprising
amino acid seguence LKKTETQ (SEQ ID NO: 1 ) or a variant
thereof .
An alternative second aspect provides a composition
comprising a peptidomimetic of a polypeptide comprising
amino acid seguence LKKTETQ (SEQ ID NO: 1 ) or a variant
thereof for treating HIV or AIDs.
An alternative second aspect provides use of a
composition comprising a peptidomimetic of a polypeptide
comprising amino acid seguence LKKTETQ (SEQ ID NO: 1 ) or a
variant thereof in the manufacture of a medicament for
treating HIV or AIDs.
In one embodiment of the first aspect the polypeptide
is LKKTETQ (SEQ ID NO :1) or Ac-LKKTETQ-OH or comprises a
fragment of thymosin beta 4 comprising LKKTETQ (SEQ ID N0:1)
or consists essentially of SEQ ID NO: 3 .
In one embodiment of the second aspect the
peptidomimetic is based on the polypeptide LKKTETQ (SEQ ID
N0:1) . In one embodiment of the second aspect the
peptidomimetic is based on a thymosin beta 4 fragment
thereof comprising LKKTETQ (SEQ ID N0:l)or a peptide
consisting essentially of SEQ ID NO: 3 .
Detailed Description
The present invention is based on the discovery that
administration of the peptide of SEQ ID NO: 1 , a peptide
derived from thymosin beta 4 (Tb4 ) , to HIV positive patients
and those that had progressed to showing symptoms of AIDS
increased CD4 and CD8 positive white blood cell counts and
decreased viral load significantly.
The peptide of SEQ ID NO: 1 has no effect on T-cell
tropic or macrophage tropic HIV isolates cultured in primary
cells, indicating its anti-HIV activity is caused by an
alternative mechanism.
Tb4 is a small naturally occurring 43 amino acid
peptide (SEQ ID NO: 2 ) first isolated from the thymus in
1981 and now identified in many tissues. Tb4 has been
implicated in many biological activities, including wound
healing, angiogenesis, actin binding, cell migration and
cell adhesion. It is anti-microbial , anti-apoptotic and has
anti-inflammatory activity.
MSDKPDMAEI EKFDKSKLKK TETQEKNPLP SKETIEQEKQ AGES (SEQ ID NO:
2 )
Until the present invention, it had not been disclosed
or suggested that LKKTETQ (SEQ ID NO: 1 ) can be used for the
treatment of HIV or AIDs.
In one embodiment the polypeptide does not comprise
full length human wild type Tb4 of SEQ ID NO: 2 .
The polypeptide for use in the first and second aspects
may include Tb4 , and/or Tb4 isoforms, analogues or
derivatives comprising SEQ ID NO: 1 or a variant thereof.
Variants of SEQ ID NO: 1 are polypeptides in which at
one or more amino acids positions there is an amino acid
insertion, deletion or substitution, either conservative or
non-conservative. There may be a mutation at one, two,
three or four of the positions of the polypeptide sequence
LKKTETQ (SEQ ID O : 1 ) .
A conservative substitution is a substitution of a
similarly sized or charged residue for another, for example
substitution of one residue in the following groups with
another in the same group would be considered conservative:
Gly, Ala
Val, lie, Leu
Asp, Glu
A s , Gin,
Ser, Thr
Lys, Arg
Phe, Tyr
In one embodiment variants are based on the consensus
sequence X L X2 X3X X X6 (SEQ ID NO: 3), wherein X is any
amino acid. Preferably X is a conservative substitution of
the native amino acid of Tb4 given in SEQ ID NO: 2 ,
In one embodiment is 1 , 2 , 3 , 4 , or 5 amino acid
residues or is absent, X2 is K , H or A , X3 is E or N , X 4 is
T or , X 5 is Q , N , E or A and X is 1 , 2 , 3 , 4 or 5 a no
acid residues or s absent.
Preferably comprises (F/L) (D/N) (S/A/T/K/N) (K/N/G)
Preferably X comprises (E/T) (K/E) (N/E) .
The polypeptide may comprise one of amino
sequences :
FDKSKLKKTETQEKN (SEQ ID NO: 4 )
FDKAKLKKTETQEKN (SEQ D NO: 5 )
FDRSKLKKTETNTEE (SEQ ID NO: 6 )
FDKTKLKKTE TQEKN (SEQ ID NO: 7 )
FDKSKLKKTNTEEKN (SEQ ID NO: 8 )
FDRSKLKKTNTEEKN (SEQ ID NO 9 )
FDKTKLKKTE AEKN (SEQ ID NO 10)
FNRAKLKKTE TQEKN (SEQ ID NO 11)
FNKAKLKKTEMQEKN (SEQ ID NO 12)
FDAKKLKHTETNEKN (SEQ ID O 13)
FNQNNLKHTE TNEKN (SEQ ID NO 14)
LDKAKLKATEMQEKN (SEQ ID NO 15)
FDKAGLKKTETEEKE (SEQ ID NO 16) .
In one embodiment the polypeptide consists essentially
of the amino acid sequence provided in SEQ ID NO: 1 or in
any one of SEQ ID NOs: 3-16.
In one embodiment the polypeptide has an N terminal
acetyl group and a C terminal hydroxy! group, for example
Ac-LKKTETQ-OH.
In another embodiment the polypeptide comprises or
consists essentially of oxidized TB4, N-terminal variants of
T j 4 , C-terminal variants of T 4 , polypeptides or peptide
fragments comprising or consisting essentially of the amino
acid sequence LKKTETQ, LKKTETQ, LKKTNTQ, KLKKTETQ, LKKTETQQ,
conservative variants thereof, or other peptide agents as
described herein, having activity as described herein.
International Application Serial No. PCT/US99/17282,
incorporated herein b y reference, discloses LKKTET (SEQ ID
NO: 17) and isoforms of Tb4 which may b e useful in
accordance with the present invention as well as amino acid
sequence LKKTETQ and conservative variants thereof, which
may b e utilized with the present invention.
International Application Serial No. PCT/GB99/00833 (WO
99/49883) , incorporated herein b y reference, discloses
oxidized Tb4 which may be utilized in accordance with the
present invention.
Although the present invention is described primarily
hereinafter th respect to SEQ D NO: 1 , t s to be
understood that the following description is intended to be
equally applicable to amino acid sequences provided as SEQ
ID NOs: 3-17 and to Tb4 and T 4 isoforms.
Many Tb4 isoforms have been identified and have about
70%, or about 75%, or about 80% or more homology to the
known amino acid sequence of Tb4 . Such isoforms include, for
example, Tb4 ΐ Tb9 , TbIO , Tb , I b12, I b13, I b14 and Tb15.
Thus, it is specifically contemplated that known Tb4
isoforms, such as Tb4 , Tb9 , TbIO , Tbΐ , Tb12, Tb13, Tb14
and Tb15, as well as Tb4 isoforms not yet identified, will
be useful in the methods of the invention. The invention
therefore further provides a composition comprising Tb4 or
fragments or variants or Tb4 isoforms T 4 , Tb 9 , O ,
Tb ΐ ΐ , I b12, Tb13, Tb14 and Tb15 or fragments or variants for
treating HIV or AIDS .
In one embodiment the composition s a pharmaceutical
composition and comprises a pharamceutically acceptable
carrier .
In another embodiment the composition is a veterinary
composition and comprises a carrier suitable for veterinary
use .
By the term "derivative thereof", we mean a peptide
within which amino acids residues are replaced by residues
(whether natural amino acids, non-natural amino acids or
amino acid mimics) with similar side chains or peptide
backbone properties.
Additionally, either one or both terminals of such
peptides may be protected by N and C-terminal protecting
groups, for example, groups with similar properties to
acetyl or amide groups. It will be appreciated that the
amino acid sequence may be varied, truncated or modified
once the final peptide is formed.
Such derivatives may increase or decrease the peptide
half-life in vivo. Examples of derivatives capable of
increasing the half-life of peptide and polypeptides
according to the invention include peptoid derivatives of
the polypeptides, D-amino acid derivatives of the
polypeptides, and peptide-peptoid hybrids.
The preparation of polypeptides is a routine process.
For example, the polypeptide can be synthesised, and many
companies offer the commercial synthesis of peptides.
Laboratory techniques are also well known for the
preparation of peptides, such methods being readily
performed by a skilled person; for example, using
recombinant DNA technologies as set out in Sambrook et al
(2001) : Molecular cloning, a laboratory manual, 3rd edition,
Cold Spring Harbor Press, Cold Spring Harbor, New York.
Hence the skilled person can prepare thymosin b 4
polypeptides using the information provided herein and from
common general knowledge.
The terms polypeptide and peptide herein are used
interchangeably and no size restriction is intended when the
term peptide is used instead of polypeptide or vice versa.
Peptides and polypeptides used according to the
invention may be subject to degradation by a number of means
(such as protease activity in biological systems) . Such
degradation may limit the bioavailability of the
polypeptides and hence the ability of the polypeptides to
achieve their biological function. There are wide ranges of
well established techniques by which derivatives that have
enhanced stability in biological contexts can be designed
and produced. Such polypeptide derivatives may have improved
bioavailability as a result of increased resistance to
protease-mediated degradation.
In one embodiment, a derivative or analogue suitable
for use according to the invention is more proteaseresistant
than the peptide from which it is derived.
The polypeptide may be made more protease-resistant by
protecting the N and/or C terminal. For example, the N
terminal may be protected by an acetyl group, or by an alkyl
or aryl group, or an alkyl-CO- or aryl-CO- group, each of
which may be optionally substituted. The C terminal may be
protected by an amide group or by a substituted amide group.
Protease-resistance of a polypeptide derivative and the
polypeptide from which it is derived may be evaluated by
means of well-known protein degradation assays. The relative
values of protease resistance for the polypeptide derivative
and polypeptide may then be compared.
Peptoid derivatives of the polypeptides of the
invention may be readily designed from knowledge of the
structure of the polypeptide. Commercially available
software may be used to develop peptoid derivatives
according to well-established protocols.
A further embodiment of a modified form of peptides
according to the invention comprises D-amino acid forms of
the peptide. The preparation of peptides using D-amino acids
rather than L-amino acids greatly decreases any unwanted
breakdown of such an agent by normal metabolic processes,
decreasing the amounts of agent which need to be
administered, along with the frequency of its
administration .
Peptides, analogues, or derivatives represent products
that may advantageously be expressed by biological cells and
the invention includes use of such agents produced
recombinant l .
The term "peptidomimetic" refers to a compound that
mimics the conformation and desirable features of a
particular peptide as a therapeutic agent, but that avoids
the undesirable features. For example, morphine is a
compound which can be orally administered, and which is a
peptidomimetic of the peptide endorphin. There are a number
of different approaches to the design and synthesis of
peptidomimetics, as is well known in the art.
The peptides can also contain further amino acid
sequences which are not derived from the amino acid sequence
of thymosin: for example, other amino acid sequences which
provide a separate function of the peptide (such as a tag,
or a catalytic domain) . Such peptides are also included in
the aspects of the invention.
Also, it will be appreciated that the invention may be
put into effect using derivatives or analogues of these
preferred peptides that still lie within the scope of the
claims with regard to SEQ ID NO: 1 and variants thereof.
All peptides, polypeptides, fragments, variants,
derivatives or peptidomimetics of Tb4 comprising or based on
SEQ ID NO :1 or a variant thereof are hereinafter referred to
as therapeutic agent (s) . Such therapeutic agents are all
capable of reducing CD4 positive cell levels in subjects
infected with HIV or AIDS.
As can be appreciated, the medicament may be
administered to a variety of different subjects. By
"subject" we include any animal that is susceptible to
developing HIV or AIDS or who is infected with the disease,
preferably a vertebrate, more preferably a mammal such as a
domesticated or farmyard animal or a human. Most preferably
the subject is a human.
The therapeutic agent may be administered orally,
topically, or parenterally in medicaments containing
conventional non-toxic pharmaceutically acceptable carriers,
adjuvants, and vehicles.
The term parenteral as used herein includes
intravenous, intraarterial, intraperitoneal, intramuscular,
subcutaneous, subconjunctival, intracavity, transdermal and
subcutaneous injection, aerosol for administration to lungs
or nasal cavity or administration by infusion by, for
example, osmotic pump.
Various means by which a medicament comprising a
therapeutic agent can be formulated are provided below.
The therapeutic agent may be formulated into
compositions having a number of different forms depending,
in particular on the manner in which the composition is to
be used. Thus, for example, the composition may be in the
form of a powder, tablet, capsule, liquid, ointment, cream,
gel, hydrogel, aerosol, spray, micelle, transdermal patch,
liposome or any other suitable form that may be administered
to a person or animal. It will be appreciated that the
vehicle for the polypeptide should be one which is well
tolerated by the subject to whom it is given, and preferably
enables delivery of the therapeutic agents to the target
cell, tissue, or organ.
Hence, it is preferred that that polypeptide is
delivered by means of a suitably protected carrier particle,
for example, a micelle.
The therapeutic agents may be used in a number of ways.
For instance, systemic administration may be required in
which case therapeutic agents may be contained within a
composition which may, for example, be ingested orally in
the form of a tablet, capsule or liquid. It is preferred
that therapeutic agents are administered by injection into
the blood stream. Injections may be intravenous (bolus or
infusion) or subcutaneous (bolus or infusion) .
Therapeutic agents may be combined in pharmaceutical
compositions having a number of different forms depending,
in particular on the manner in which the composition is to
be used. Thus, for example, the composition may be in the
form of a powder, tablet, capsule, liquid, ointment, cream,
gel, hydrogel, aerosol, spray, micelle, transdermal patch,
liposome or any other suitable form that may be administered
to a person or animal. It will be appreciated that the
vehicle used to provide the treatment should be one which is
well tolerated by the subject to whom it is given, and
preferably enables delivery of the therapeutic to the target
cell, tissue, or organ.
In a preferred embodiment, the pharmaceutical vehicle
is a liquid and the pharmaceutical composition is in the
form of a solution. In another embodiment, the
pharmaceutical vehicle is a gel and the composition is in
the form of a cream or the like.
Therapeutic agents may also be incorporated within a
slow or delayed release device. Such devices may, for
example, be inserted on or under the skin, and the
therapeutic agent may be released over weeks or even months.
Such devices may be particularly advantageous when long term
treatment with the therapeutic agents is required and which
would normally require frequent administration (e.g. at
least daily injection) .
It will be appreciated that the amount of a therapeutic
agent that is required is determined by its biological
activity and bioavailability which in turn depends on the
mode of administration, the physicochemical properties of
the therapeutic agent employed, and whether the therapeutic
agent is being used as a mono-therapy or in a combined
therapy. Also, the amount will be determined by the number
and state of target cells to be treated. The frequency of
administration will also be influenced by the abovementioned
factors and particularly the half-life of the
therapeutic agent within the subject being treated.
Optimal dosages to be administered may be determined by
those skilled in the art, and will vary with the particular
therapeutic agent in use, the strength of the preparation,
the mode of administration, and the advancement of the
disease condition. Additional factors depending on the
particular subject being treated will result in a need to
adjust dosages, including subject age, weight, gender, diet,
and time of administration.
Known procedures, such as those conventionally employed
by the pharmaceutical industry (e.g. in vivo
experimentation, clinical trials, etc.), may be used to
establish specific formulations of therapeutic agents and
precise therapeutic regimes (such as daily doses of the
therapeutic agents and the frequency of administration) .
Daily doses may be given as a single administration
(e.g. a single daily injection).
Alternatively, the therapeutic agents used may require
administration twice or more times during a day. As an
example, an therapeutic agents may be administered as two
(or more depending upon the severity of the condition) daily
doses of between 1 g and 1000 g (i.e. assuming a body
weight of 70kg) . A patient receiving treatment may take a
first dose upon waking and then a second dose in the evening
(if on a two dose regime) or at 3 or 4 hourly intervals
thereafter. Alternatively, a slow release device may be used
to provide optimal doses to a patient without the need to
administer repeated doses.
In one embodiment the medicament comprising the
therapeutic agent is administered subcutaneously or is
formulated for subcutaneous administration to a subject. In
one embodiment the medicament is an injectable composition.
In one embodiment the medicament is formulated to
provide between 1 to 1000 mg of the therapeutic agent per
administration .
A fifth aspect of the invention provides a population
of stem cells that secrete thymosin b4 for use as a
medicament for the prevention or treatment HIV or AIDS.
Until the present disclosure, it had not been disclosed or
suggested that such stem cells would have this utility.
Stem cells are cells that have the potential to
differentiate into a number of cell types in the body.
Theoretically, stem cells may divide without limit to
replenish other cells for as long as the organism is alive.
Upon differentiation, the daughter cell has the potential to
remain a stem cell or become another cell type, for example
lung cell and display its characteristics, thus holding
promise for many 5 diseases by replacing damaged tissues.
These phenomena may be induced under specific physiological
and experimental conditions.
In general, stem cell therapy represents a therapeutic
method by which degenerative and/progressive diseases (such
as those caused by premature death or malfunction of cell
types that the body is unable to replace) may be treated. It
is hoped that addition of stem cells may help nucleate and
promote the development of functional cells and/or tissues
to replace those lost, thereby restoring normal healthy
activity/ function .
For the purposes of the present invention, "stem cells"
are taken to comprise nullipotent, totipotent or pluripotent
cells, and progenitor cells (or precursor cells) to comprise
multipotent cells. For the avoidance of doubt, the
medicament and methods of the invention can comprise a
therapeutically effective quantity of either stem or
progenitor cells, or both stem and progenitor cells.
A suitable source of stem cells that may be used in
accordance with the present invention are cells derived from
the inner cell mass/epiblast of pre-implantation embryos.
Such embryonic stem (ES) cells are readily obtainable and
are capable of giving rise to all possible embryonic and
adult cell lineages. In particular, the undifferentiated
human ESC (HI line from WiCell Research Institute, Inc,
Madison, W I : www.wicell.org) could be used in the invention;
this cell line is commercially available.
A further source of stem cells that can be used in the
present invention are umbilical cord-derived cells. A still
further source of stem cells, are those isolated from adult
tissues, including adipose tissues.
Where the medicament of the invention involves the use
of biological cells, preferably the formulation for
comprises biological cells in a suitable liquid carrier.
Such a liquid carrier is preferably non-immunogenic, and may
comprise a saline solution, cell culture medium, or
distilled water. Formulations for injection may be as
described above, or may also be provided in the form of a
gel, which may preferably be capable of resolution by the
body of the subject treated. Formulations suitable for
implantation may take the forms described for injection or
inhalation, and may also comprise biological cells provided
in a scaffold or matrix capable of providing a foundation
for new tissue development.
The methods of the invention are especially useful for
the treatment of AIDS and related clinical conditions such
as AIDS related complex (ARC) , progressive generalized
lymphadenopathy (PGL) , Kaposi's sarcoma, thromobocytopenic
purpura, AIDS -related neurological conditions such as AIDS
dementia complex, multiple sclerosis or tropical
paraperesis, anti-HIV antibody-positive and HIV-positive
conditions, including such conditions in asymptomatic
pat en s ,
As well as treating AIDS or HIV the invention can also
be used to treat or prevent the symptoms or effects of a
viral infection in an infected patient. In one embodiment
the viral infection is a retroviral infection, in particular
an HIV infection.
The treatment n accordance with the present invention
may be supplemented with treatment with another therapeutic
agent. The treatments may be administered simultaneously
(i.e., concurrently) in either the same or different
pharmaceutical compositions or sequentially in any order.
The amounts of the active ingredient (s) and pharmaceutically
active agent (s) and the relative timings of administration
wil be selected in order to achieve the desired combined
therapeutic effect.
Examples of other therapeutic agents include nucleotide
reverse transcriptase inhibitors such as zidovudine,
didanosine, lamivudine, zalcitabine, abacavir, stavidine,
adefovir, adefovir dipivoxil, fozivudine, todoxil,
emtricitabine, alovudine, amdoxovir, elvucitabine, and
similar agents; non-nucleotide reverse transcriptase
inhibitors (including an agent having anti- oxidation
activity such as iramunocal, oltipraz, etc.) such as
nevirapine, delavirdine, efavirenz, loviride, immunocal,
oltipraz, capravirine, T C-278 , TMC-125, etravirine, and
similar agents; protease inhibitors such as saquinavir,
r tonav i , indi avir, ne fi a i , amp re av , fosa p enav ,
brecanavir, raltegravir, atazanavir, tipranavir, palinavir,
lasinavir, and similar agents; entry inhibitors such as
enfuvirtide (T-20), T-1249, PRO-542, PRO-140, TNX-355, BMS-
80 6 , 5-Helix and similar agents; lntegrase inhibitors such
as L-870, 810, raltegravir and similar agents; Budding
inhibitors such as PA--344 and PA--457, and similar agents;
and CXCR4 and/or CCR5 inhibitors such as vicriviroc (Sch-C) ,
Sch-D, TAK779, maraviroc (UK 427,857), TAK449 and similar
agents .
In one embodiment the method comprises administering
the therapeutic agent a subject being treated with another
therapeutic agent as listed above or otherwise.
In one embodiment the therapeutic agent is for
administering to a subject being treated with another
therapeutic agent as listed above or otherwise.
In an embodiment the therapeutic agent and other
therapeutic agent act synergistically in the treatment of
HIV or AIDS.
The terms "treating" and "treatment" as used herein refer
to reduction in severity and/or frequency of symptoms,
elimination of symptoms and/or underlying cause, prevention
of the occurrence of symptoms (prophylaxis) and/or their
underlying cause, and improvement or remediation of damage.
Thus, for example, the present method of "treating" a
disorder encompasses both prevention of the disorder in a
predisposed individual and treatment of the disorder in a
clinically symptomatic individual.
"Treating" as used herein covers any treatment of, or
prevention of a condition in a vertebrate, a mammal,
particularly a human, and includes: inhibiting the
condition, i.e., arresting its development; or relieving or
ameliorating the effects of the condition, i.e., cause
regression of the effects of the condition.
"Prophylaxis" or "prophylactic" or "preventative" therapy
or "prevent" or "prevention" as used herein includes
preventing the condition from occurring or ameliorating the
subsequent progression of the condition in a subject that
may be predisposed to the condition, but has not yet been
diagnosed as having it.
In one embodiment the method or composition prevents or
slows progression from HIV infection to AIDS. In another
embodiment the method or composition lessens the symptoms of
AIDS. In another embodiment the method or composition
improve immune response in subjects with AIDS. In another
embodiment the method or composition improve quality of life
for subjects with AIDS. In another embodiment the method or
composition prolong life span in subjects with AIDS.
Throughout this specification, unless the context
requires otherwise, the word "comprise", or variations such
as "comprises" or "comprising", will be understood to imply
the inclusion of a stated element or integer or group of
elements or integers but not the exclusion of any other
element or integer or group of elements or integers.
It must also be noted that, as used in the subject
specification, the singular forms "a", "an" and "the"
include plural aspects unless the context clearly dictates
otherwise .
It will be apparent to the person skilled in the art
that while the invention has been described in some detail
for the purposes of clarity and understanding, various
modifications and alterations to the embodiments and methods
described herein may be made without departing from the
scope of the inventive concept disclosed in this
specification .
The invention is now further described in detail by
reference to the following example. The example is provided
for purposes of illustration only, and is not intended to be
limiting unless otherwise specified. Thus, the invention
encompasses any and all variations which become evident as a
result of the teaching provided herein.
Example 1 :
LKKTETQ (SEQ ID NO :1) is a fragment of Thymosin B4
(Tb4 ) .
The peptide Ac-LKKTETQ-OH was the active therapeutic
agent investigated in a pilot trial on twenty similar HIV
positive men and women, some who had progressed to severe
AIDS .
Each man or women was injected subcutaneously with a
composition comprising the peptide Ac-LKKTETQ-OH (lOmg),
once a week for 12 weeks. Blood tests for CD4 and CDspositive
cells and HIV viral load (by PCR) and routine
pathology were conducted at baseline, 8 weeks and 12 weeks.
The results are displayed in the three Tables below.
Hemoglobin levels, Creatinine, ALT, AST, Alkaline Phos,
Bilirubin, GGT, Albumin, Total Protein and globulin tests
did not show anything remarkable. A small drop in blood
glucose was seen but this was not determined to be
significant. No adverse effect has been reported so far.
Table 1 . WEIGHT
Patient Sex Weiqht
Initial 8 weeks 12 weeks
A M 44 55 58
B F 67 70 70.5
C M 74 78 78.5
D F 75 78 78.5
E F 88 92 92
F M 47 52 52
G M 52 59 72
H M 66 7 1 72
1 F 78 82 82
J F 76 79 79
K F 67 72 7 1
L F 60 60.5 7 1
M F 48 50.5 5 1
N M 48 58 65
0 F 7 1 72 72
P F 78 79 80
Q F 69 7 1.5 72
R M 67 68.5 69
S F 58 6 1 60
T F 55 56 55
CD4 -positive and CD8 -positive white blood cell
Patient CD4
Initial 8 weeks 12 weeks
A 110 300 430
B 262 3 16 416
C 496 602 675
D 732 8 16 879
E 146 4 15 604
F 96 158 175
G 130 407 425
H 292 456 625
1 383 477 662
J 126 348 404
K 127 381 498
L 169 439 510
M 317 301 424
N 119 335 452
0 595 543 749
P 595 428 616
Q 269 466 602
R 249 259 368
S 200 471 469
T 196 466 571
Patient CD8
Initial 8 weeks 12 weeks
A 10 1 980 1330
B 318 360 459
C 1059 131 0 171 8
D 688 1379 1394
E 321 1044 1339
F 1310 1296 1226
G 208 959 1190
H 1031 2 120 2647
1 759 866 958
J 652 773 800
K 278 594 712
L 621 893
M 786 606 983
N 245 939 1034
0 6 13 617 880
P 612 648 890
Q 247 456 521
R 6 18 1108 1384
S 323 1125 1201
T 407 594 660
Table 3 : HIV Viral Load
Patient HIV Viral Load (copies/ml)
Initial 8 weeks 12 weeks
A 214000 200 undetectable
B 50809 98687 5151
C 120 24 undetectable
D 101 30 undetectable undetectable
E 223767 undetectable undetectable
F 256228 2841 0 undetectable
G 257000 450 undetectable
H 114594 8214 undetectable
1 59509 1761 6 undetectable
J 38988 undetectable undetectable
K 128685 15201 undetectable
L 53814 undetectable undetectable
M 39786 undetectable undetectable
N 144000 145 undetectable
0 13803 undetectable undetectable
P 5060 undetectable undetectable
Q 3567 undetectable undetectable
R 52915 undetectable undetectable
S 14921 2 12004 undetectable
T 115281 8781 undetectable
Example 2 :
The peptide Ac-LKKTETQ-OH was evaluated in fresh human
peripheral blood mononuclear cells (PBMCs) against CXCR4
coreceptor-trophic HT/92/599 and the CCR5 coreceptor-trophic
BaL subtype B strains of HIV-1. The peptide was added to
the cells at 8 hours prior to infection and immediately
prior to infection. The AZT control compound was evaluated
in parallel with the peptide and yielded EC5 0 values of 8
and 6 nM against HI V -1 T/ 92 / 5 99 and values of 6 and 10 nM
against HIV-l BAL . The peptide of SEQ ID NO :1 did not
demonstrate antiviral activity in the PBMC assays when
evaluated at concentrations up to a high test concentration
of 500 mM.
The results of the pilot trial were very encouraging.
The key parameters of HIV viral load, CD4 positive and CD8
positive white cell blood counts, and weight gain have all
shown extremely significant improvements from the baseline -
perhaps the most significant is the HIV viral load which, in
one patient, has gone from levels of 257000 to undetectable
over the 12 week period of the pilot trial.
CLAIMS :
1 . A method of treating HIV or AIDS comprising
administering a composition comprising a polypeptide
comprising LKKTETQ (SEQ ID NO: 1 ) or a variant thereof.
2 . A method of treating HIV or AIDS comprising
administering a composition comprising a peptidomimetic of a
polypeptide comprising amino acid sequence LKKTETQ (SEQ ID
NO: 1 ) or a variant thereof.
3 . The method of claim 1 in which the polypeptide is
LKKTETQ (SEQ ID NO :1) or Ac-LKKTETQ-OH .
4 . The method of claim 1 in which composition comprises
thymosin beta 4 or a fragment thereof comprising LKKTETQ
(SEQ ID NO:l) or SEQ ID NO: 3 .
5 . The method of claim 2 in which the peptidomimetic is
based on the polypeptide LKKTETQ (SEQ ID NO:l) or SEQ ID NO:
3 .
6 . The method of claim 2 in which the peptidomimetic is
based on thymosin beta 4 or a fragment thereof comprising
LKKTETQ (SEQ ID NO :1).
7 . The method of claim 1 for treating AIDS, AIDS related
complex (ARC) , progressive generalized lymphadenopathy
(PGL) , Kaposi's sarcoma, thromobocytopenic purpura, AIDSrelated
neurological conditions such as AIDS dementia
complex, multiple sclerosis or tropical paraperesis, antiHIV
antibody-positive a d HIV-positive conditions, including
such conditions in asymptomatic patients.
8 . The method of claim 1 for treating the symptoms or
effects of a viral infection in an infected patient,
9 . The method of claim 8 in which the viral infection is a
retroviral infection.
10 . The method of claim S , in which the retroviral
infection is an HIV infection.
11. The method of claim 1 in which the composition is
administered to a subject being treated with another
therapeutic agent .
12. The method of claim 11 in which the other therapeutic
agent is a nucleotide reverse transcriptase inhibitor, a
non-nucleotide reverse transcriptase inhibitors, a protease
inhibitor, an entry inhibitor, an integrase inhibitor, or a
budding inhibitor.
13. The method of c la im 1 wherein the sub ect is being
treated with a plurality of other therapeutic agents.