Title: Peptide T and related peptides in the treatment
of inflammation, including multiple sclerosis
United States Patent: 6,265,374
Inventors: Andersen; Anders Jorgen (Kokkedal, DK); Aston;
Roger (Wiltshire, GB); Carlen; Peter Louis (Ontario, CA); Doob; Penelope
Reed (Ontario, CA); MacFadden; Douglas Kevin (Ontario, CA); Phipps; David
James (Ontario, CA); Rathjen; Deborah (New South Wales, AU); Widmer; Fred
(New South Wales, AU)
Assignee: Advanced Immuni T, Inc. (Stonybrook, NY)
Appl. No.: 421845
Filed: October 20, 1999
A method of treating inflammation in patients in need of such treatment
by administering an effective amount of I-A-B-C-D-E-F-G-H-II (General
Formula), wherein A is Ala, Gly, Val, Ser, Thr or absent, B is Ala, Gly,
Val, Ser, Thr, or absent, C is Ser, Thr or absent, D is Ser, Thr, Asn, Glu,
Arg, Ile, Leu or absent, E is Ser, Thr, Asp or absent, F is Ser, Thr, Asp
or absent, G is Tyr or absent, H is Thr, Arg, Gly, Met, Met(O), Cys, Thr,
Gly or absent and I is Cys or absent II is Cys, an amide group, an ester
group or absent. At least one of the amino acids optionally is substituted
by a monomeric or polymeric carbohydrate or derivative thereof, such
substitution being accomplished through hydroxyl and/or amino acid and/or
amido groups of the amino acids, and wherein the peptide composes at least
4 amino acid residues, and a pharmaceutically acceptable salt thereof.
Description of the Invention
The present invention relates, broadly to the treatment or
prevention of inflammation, whether caused by bacteria, viruses and/or
other infective agents, opportunistic infections (which may be consequent
on an immunodepressed state, for example resulting from cancer or therapy,
particularly cytotoxic drug therapy or radiotherapy) autoimmunity or
otherwise. In particular embodiments, the invention relates to the
prevention or treatment of neurodegenerative or demyelinating diseases
such as HTLV-1-associated myelopathy (HAM), multiple sclerosis (MS) and
symptoms or diseases in humans which are associated with chronic immune
activation. The invention also relates to pharmaceutical compositions
useful in such treatment and/or prevention and to certain active peptides
Septic shock is an illustration of a disease involving inflammation. Many
of the clinical features of Gram-negative septic shock may be reproduced
in animals by the administration of lipopolysaccharide (LPS). The
administration of LPS to animals can prompt severe metabolic and
physiological changes which can lead to death. Associated with the
injection of LPS is the extensive production of tumour necrosis factor
alpha (TNF-.alpha.). Mice injected with recombinant human TNF develop
piloerection of the hair (ruffling), diarrhoea and a withdrawn and unkept
appearance, followed by death if sufficient amounts are given. Rats
treated with TNF become hypotensive, tachypneic and die of sudden
respiratory arrest (Tracey et al. 1986 Science 234, 470). Severe acidosis,
marked haemoconcentration and biphasic changes in blood glucose
concentration were also observed.
Histopathology of such rats revealed severe leukostatsis in the lungs,
haemorraghic necrosis in the adrenals, pancreas and other organs and
tubular necrosis of the kidneys. All of these changes were prevented if
the animals were pretreated with a neutralizing monoclonal antibody
The massive accumulation of neutrophils in the lungs of TNF-treated
animals reflects the activation of neutrophils by TNF. TNF causes
neutrophil degranulation, respiratory burst as well as enhanced neutrophil
antimicrobacterial and anti-tumor activity (Klebanoff et al, 1996, J.
Immunol. 136, 4220; Tsujimoto et al, 1986 Biochem. Biophys. Res. Commun.
137, 1094). Endothelial cells are also an important target for the
expression of TNF toxicity. TNF diminishes the anticoagulant potential of
the endothelium, inducing procoagulant activity and down-regulating the
expression of thrombomodulin (Stern and Nawroth, 1986 J. Exp. Med. 163,
TNF is a product of activated macrophages and is produced in response to
infection and malignancy. It was first discovered in LPS-treated mice as a
serum factor which caused the haemorraghic necrosis of transplanted tumour
cells in culture (Carswell et al, 1975 PNAS 72, 3666; Helson et al, 1975,
Nature 258, 731). Cachexia, which is characteristic of chronic exposure to
TNF, it a common symptom of advanced malignancy and severe infection. It
is characterized by abnormal lipid metabolism with hypertriglyceridaemia.,
abnormal protein and glucose metabolism and body wasting. Chronic
administration of TNF and IL-1 in mice, rats and/or humans causes
anorexia, weight loss and depletion of body lipid and protein within 7 to
10 days (Cerami et al, 1985, Immunol. Lett. 11, 173; Fong et al, 1989 J.
Exp. Med. 170, 1627, Moldawer et al, Am. J. Physiol., 254 G450-G456, 1988;
Fong et al, Am. J. Physiol. 256 R659-R665 (1989); McCarthy et al, Am. J.
Clin. Nutr. 42 1179-1182, 1982). TNF levels have been measured in patients
with cancer and chronic disease associated with cachexia. The results are
inclusive since large differences in TNF levels have been reported. These
may have been explicable by the short half-life of TNF (6 minutes),
differences in TNF serum binding protein or true differences in TNF levels
in chronic disease states.
TNF-.alpha. and IL-1, with their common functional activities such as
pyrogenicity, somnogenicity and being mediators of inflammation, have been
impacted in the pathology of other diseases associated with chronic
inflammation. apart form toxic shock and cancer-related cachexia. TNF has
been detected in synovial fluid in patients with both rheumatoid and
reactive arthritis (Saxne et al, 1988, Arthrit. Rheumat. 31, 1041). Raised
levels of TNF have been detected in renal transplant patients during acute
rejection episodes (Maury and Teppo 1987, J. Exp. Med. 166, 1132). In
animals, TNF has been shown to be involved in the pathogenesis of
graft-versus-host disease in skin and gut following allogenic marrow
Administration of a rabbit anti-murine TNF antibody was shown to prevent
the histological changes associated with graft-versus-host disease and to
reduce mortality (Piquet et al, 1987, J. Exp. Med. 166, 1220). TNF has
also been shown to contribute significantly to the pathology of malaria
(Clark et al, 1987, Am. J. Pathol. 129, 192-199). Further, elevated serum
levels of TNF have been reported in malaria patients (Scuderi et al, 1986,
Lancet 2, 1364-1365).
Multiple sclerosis (MS) is generally considered by many authorities to be
a chronic inflammatory disease.
Both MS and HTLV-1 associated myelopathy (HAM) affect the central and the
peripheral nervous systems and both may present clinically as a myelopathy
affecting both the spinal nerves and the spinal myelinated nerve fibres.
Multiple sclerosis (MS) is a chronic demyelinating disease of the central
nervous system and is the commonest chronic neurological disease of young
adults. The incidence of MS and its pattern of distribution have been
unchanged for decades. The disease remains essentially untreatable.
MS has always been regarded as a disease of the temperate zones and has a
prevalence in the northern United States, Canada and Europe of 1:1000. The
disease has a gender predilection of 1.5:1 (female:male).
MS usually affects multiple areas of white matter in the central nervous
system (CNS), most frequently, the perventricular white matter, brainstem,
spinal cord and the optic nerves. The primary process destroys myelin
sheaths and eventually kills oligodendrocytes creating the characteristic
plaque of MS.
The early development of the plaque is characterised by the development of
perivascular inflammation followed by the migration of lumpnocytes, plasma
cells and macrophages into the lesion. This is followed by astrocyte
gloisis and the attempts of remyelination by oligodendrocytes. The plaque
is surrounded by lumphocytes.
Although the aetiology of MS is still unknown, the focus of research
efforts that have led to plausible hypotheses have been those of immune
dysregulation including autoimmunity and genetic predisposition, both of
which may play a role in the actual development of disease.
Multiple immunological abnormalities are reproducibly found in patients in
the acute stage of the disease. The synthesis of immunoglobins, although
normal in the periphery, is increased in the central nervous system and
the antibodies produced have a characteristic banding pattern. The
antigenic specificity of these antibodies is not known and it is unclear
whether they have a role to play in the progression of disease.
Various stressors known to activate the immune system such as viral
infection or surgery can also produce an exacerbation of MS. Other
activators such as .gamma.-interferon produce similar effects when
administered. In addition, immunosuppresive anti-inflammatory therapy with
corticosteroids for example, can produce modest remission or at least
palliation for short periods of time, although this therapy is
Lymphocyte reactivity against two neuronal antigens myelin basic protein
and proteolipid has been demonstrated. Although not proven, this activity
would form the basis for an autoimmune response against neuronal tissue.
The discovery of the neurotropic capacity of HTLV-1 in patients form
Martinique with tropical spastic paraparesis (TSP) and in Japan with
chronic myleopathy, has demonstrated HTLV-1 as the common aetiologic agent
of these diseases. It has subsequently been shown that the neurologic
manifestations of HTLV-1 infection are the same despite the varied
geographic regions in which they are described.
The neurological signs of this chronic retroviral infection including
slowly progressive spastic paraparesis with spastic bladder and minimal
sensory deficits result form involvement of the pyramidal tracts in a
bilateral and symmetrical fashion predominantly at the thoracic level of
the spinal cord.
The peripheral nervous system has been shown to be involved, resulting in
slowing of nerve conduction velocities in than lower limbs. Systemic
manifestations of HTLV-1 in patients with HTLV-1 myelopathy have been
described and include inflammatory involvement of the lungs, skin, eyes
and striated muscle producing a myositis. In addition, patients experience
profound fatigue similar to MS. The clinical manifestations of the disease
are very similar to MS and are frequently confused with the latter.
There are at least four possible pathogenetic mechanisms whereby HTLV-1
can involve the CNS to produce HAM. These may include a slow virus
infection, a cell-mediated immune response and a predominantly humoral
immune mediated mechanism and the development of an autoimmune phenomenon.
The slowly progressive course supports the hypothesis of a slow virus
infection. The finding of peri-vascular cuffing in post-mortem specimens
as well as transiently favourable response to steroids supports the
hypothesis that an inflammatory immune reaction, probably a result of
viral infection, is responsible for the development of HAM.
These two diseases have many similarities and dissimilarities, both
clinical and neurological. Both diseases are a form of demyelinating
disease whereby the myelin sheath of the nervous system is destroyed by
one of many mechanisms common to both diseases and also peculiar to either
of the diseases. MS is a multi-faceted disease in that can be both a
central nervous system disease which can include a myelopathy. Conversely,
HTLV-1-associated myelopathy is predominantly a myelopathy which can
occasionally demonstrate central nervous system effects. Furthermore, MS
can affect the peripheral nervous system in ways that are common HTLV-1.
Myelopathy, as already mentioned in being a disorder of the spinal cord,
can have many different aetiologies from MS and HAM. Various forms of
myelopathy, most of which are mediated by inflammation include the
B12 or folate deficiency;
motor neuron disease;
spinal cord compression from tumour, disc or arthritis;
lupus erythematosus of the spinal cord; and
Chronic inflammation or, as more commonly known, chronic immune system
activation occurs in response to persistent antigen whose origin may be
exogenous or endogenous or may result from an autoimmune state. Such
chronic inflammation results in local tissue destruction and depending
upon the type of inflammation can result in systemic effects due to the
sustained production of inflammatory mediators. Such inflammatory
mediators include the cytokines which are soluble mediators produced by
activated lymphocytes and macrophages and effect cellular communication
and physiological response. Chronic immune activation can occur as a
result of infectious diseases, such as chronic fatigue syndrome or toxic
shock syndrome or through autoimmune mechanisms resulting in such
conditions as rheumatoid arthritis, inflammatory bowel disease and
variants such as graft versus host interactions.
The immune response to antigen may be divided into four overlapping
phases: initiation (antigen presentation), amplification (cell
activation), effector and regulation (Roitt et al, "Immunology",
Gower Medical Publ. London. EK, 1989; "Basic and Clinical Immunology,
Stites et al, Eds, Appleton and Lance, Norwalk, Conn., 1991). Briefly
antigen is phagocytosed by antigen presenting cells (APC) which must
express major histocompatibility (MHC) Class II molecules on their
surface. In this respect, cells of the macrophage/monocyte lineage (CD4
positive) and B cells (CD4 negative) may act as APC. Following
phargocytosis, antigen is processed intracytoplasmically and expressed on
the surface as antigenic fragments in association with MHC-II molecules.
The combination of antigen/MHC-II induces the activation of T helper cells
(CD4 positive) in an antigen-specific manner and primes them to receive a
second antigen non-specific activating signal. Activated T helper cells
than induce the activation of effector T cells (cytoxic lymphocytes, CD4
negative) and B cells which produce antibody. Effector cells and molecules
facilitate the elimination of antigen by a variety of antigen specific and
non-specific mechanisms that may result in host tissue damage if effector
mechanisms are expressed inappropriately by deposition of antigen or
immune complexes on host tissues, responding to self-antigens or as a
result of a prolonged (chronic) immune response. Regulation of the immune
response via removal of antigen, active suppression or idiotypic
regulation limits the normal immune response to a duration of one to three
Chronic fatigue syndrome (CFS) or chronic fatigues immune dysfunction
syndrome (DeFritas et al, Proc. Natl. Acad. Sci. 88, 2922-2926 (1991)) is
a condition of unknown aetiology characterised by a diverse set of signs
and systems including severe fatigue, post-exertional malaise, headaches,
night sweats, myalgia, ataxia, low grade fever and lymphadenophathy (CDWR
1-3: Joncas J H. Welcome and Introduction in: Proceedings of a Workshop:
Chronic Fatigue Syndrome, Can. Dis. Weekly Report vol. 17S1E, January 1991
pages 1-3. ).
Although the origin of CFS is unknown its symptoms are consistent with
over-production of cytokines (Landay et al, Lancet 338 707-712 (1991)).
CFS-like symptoms have been observed following the therapeutic
administration of interferons (INFs) (Lloyd et al, Med. J. Aust. 151
122-124 (1989); Lever et al, Lancet 2 101 (1988); Mowbray et al, Br. Med.
Bull. 47 886-894 (1991)) and interleukin-2 (IL-2) (Cheney et al, Annal.
Intern. Med. 110 321 (1989)). In a trial of IFN-.alpha. in patients with
CFS, the drug exacerbated the condition further supporting a
cytokine-mediated pathogenesis (McBride et al, Br. Med. Bull. 47 895-907
)1991)). The serum and cerebrospinal fluid of patients with CFS has been
shown to contain increased levels of IL-2, IFN and IL-1 (Wallace et al,
Arth. Rheum. 32 1334-1335 (1989); Shepperd The Practitioner 233 41-46
(1989)); as well as IL-6 (Chao et al, J. Infect. Dis. 162 1412 (1990)). In
addition neopterin, a marker of machrophage activation (Chao et al, J.
Infect. Dis. 162 1412 (1990)). and the IFN-associated enzyme
2'-5'-oligoadenylate synthetase (Klimas et al, J. Clin. Microbiol. 28
1403-1410 (1990)) are both incresed in CFS as are other markers of
macrophage activation such as ICAM-1 and LFA-1 (Gupta et al, Scand. J.
Immunol. 33 319-327 (1991)). CFS-associated anery to skin test antigens
(Johnson et al. FASEB J. 5 2706-2712 (1991)), reduction in lymphocyte
response to mitogens (Klimas et al, J. Clin. Microbiol. 28 1403-1410
(1990)) and soluble antigens (Gupta et al, Scand. J. Immunol. 33 319-327
(1991)) are consistent with macrophage dysfunction (Prieto et al, Scand.
J. Immunol. 30 13-20 (1989)) and may be explained by an autocrine
exhaustion of immunocompetent cells by chronic activation resulting in
immunodysregulation (CDWR 1-3: Joncas J H, Welcome and Introduction in:
Proceedings of a Workshop Chronic Fatigue Syndrome. Can. Dis. Weekly
Report vol. 17S1E, January 1991, pages 49-50).
CFS and the acquired immune deficiency syndrome (AIDS, see below) share
many symptoms (Miller et al, Neurology 41 1603-1607 (1991)) and laboratory
findings (Gupta et al, Scand. J. Immunol. 33 319-327 (1991)); and one
study has demonstrated an association between infection with the HIV
related retrovirus HTLV-II and CFS (Gupta et al, Scand. J. Immunol. 33
There is no accepted drug therapy for CFS. There have been anecdotal
reports of beneficial effects following administration of amantidine,
monoamine oxidase inhibitors (ie phenelzine), fatty acid supplements
(McBride et al, Br. Med. Bull. 47 895-907 (1991)) and 5-hydroxytryptophan
(Caruso et al, J. Int. Med. Res. 18 201-209 (1990)) among others, but
controlled studies have not demonstrated efficacy.
Toxic shock syndrome (TSS) is produced by a Staphylococcus aureus
enterotoxin, toxic shock syndrome toxin-1 (TSST-1). TSST-1 belongs to a
family of staphylococcal exterotoxins which are mitogenic for T cells
expressing particular V.beta. genes (Kappler et al, Science 244 811-813
(1989)). As a result of their non-specific mitogenicity, staphylococcal
enterotoxins can induce the proliferation of up to 20% of T-cells and have
been called "superantigens" (Johnson et al, Sci. Am. 266 92-101
(1992)). Macrophages are required to present TSST-1 to T-cells (Poindexter
et al, J. Infect. Dis. 151 65-72 (1985)); however, like other
staphylococcal enterotoxins, TSST-1 does not require antigen processing
for T-cell activation (Pontzezr et al, Proc. Natl. Acad. 88 125-128
(1991)). The native molecule binds outside the antigen binding cleft of
MHC Class II molecules to a non-polymorphic region of the .beta. chain
(Fraser Nature 339 221-223 (1989); Johnson et al, FASEB J. 5 2706-2712
The symptoms of TSS (such as fever, rash, hypotension, nausea, vomiting
and diarrhoea) are consistent with over-activation of the immune system
(Johnson et al, Sci. Am. 266 92-101 (1992)) and over-production of
cytokines (Ikejima et al, J. Clin. Invest. 73 1312-1320 (1984); Micussan
et al, Immunology 58 203-208 (1986)). These symptoms have been reproduced
in animal models by the administration of tumour necrosis factor (TNF) (Miethke
et al. J. Exp. Med. 175 91-98 (1992)). Massive immune activation of this
nature could lead to exhaustion of immunocompetent cells and may explain
the immunosuppression associated with enterotoxin shock (Langford et al,
Infect. Immun. 22 62-68 (1978)) and in vitro enterotoxin-induced T-cell
anergy (O'Hehir et al, Immunol. Lett. 30 163-170 (1991)).
Therapy of TSS involves immediate replacement of lost fluid to counter
hypovolaemia. If the patient fails to respond to anti-staphylococcal
antibiotics than steroid therapy (ie methylprednisone) may be required for
those in severe shock (Todd, Drugs 39 856-861 (1990)).
Rheumatoid arthritis (Marrow et al, "Autoimmune Rheumatic
Disease", Blackwell Scientific Publ. Oxford UK, Chapter 4, pp148-207
(1987)) is a disease characterised by chronic inflammation and erosion of
joints that may affect up to 3% of the population, including children.
Symptoms of rheumatoid arthritis include morning stiffness, swelling and
pain upon motion in at least one joint and joint swelling. Non-specific
symptoms including lethargy, anorexia and weakness as well as fever and
lymphadenopathy (characteristic of immune activation) may antedate joint
involvement. Extra-articular manifestations of rheumatoid arthritis
include vasculitis, cataracts, uveitis, interstitial fibrosis,
pericarditis and myocarditis, pheripheral neuropathy, myeloid deposits,
chronic anaemia and subcutaneous and pulmonary nodules.
Genetic factors and infectious agents including bacteria, fungi,
mycoplasmas and viruses have been associated with the development of
rheumatoid arthritis. Mild rheumatoid arthritis may be treated with
non-steroidal anti-inflammatory drugs while severe cases require sytemic
coricosteroids, anti-metabolites or cytotoxic agents. Experimentally,
anti-CD4 monoclonal antibodies have been used to treat rheumatoid
arthritis (Horneff et al, Cytokine 3 266-267 (1991); Horneff et al, Arth.
Rheum. 34 129-140 (1991) and Shoenfeld et al, Clin. Exp. Rheum. 9 663-673
Inflammatory bowel disease (IBD) is a chronic inflammatory condition that
fulfills some of the criteria of an autoimmune disease (Snook, Gur 31
961-963 (1990)). Inflammation and tissue damage involes the recruitment
and activation of neutrophils, macrophages and lymphocytes (MacDermott et
al, Adv. Immunol. 42 285-328 (1988)) which generate cytokines and
proinflammatory molecules such as prostaglandins and leukotrienes (MacDermott.
Mt. Sinai J. Med. 57 273-278 (1990)). As a result of chronic activation of
immunocompetent cells, IL-1, IL-6 (Starter. Immunol. Res. 10 465-471
(1991); Fiocchi, Immunol. Res. 10 239-246 (1991)) and TNF-.alpha. (MacDermott,
Mt. Sinai J. Med. 57 273-278 (1990)) are all elevated in IBD patients.
Drugs used to treat IBD include anti-inflammatory agents such as
sulphasalazine (5-ASA), corticosteroids, cyclosporin A and azathioprine (Hanauer,
Scand. J. Gastroenterol. 25 (Suppl 175) 97-106 (1990); Peppercorn, Annal.
Intern. Med. 112 50-60 (1990)). Experimentally, anti-CD4 monoclonal
antibodies have been used to successfully treat ulcerative colitis (Emmrich
et al, Lancet 338 570-571 (1991)).
While a host may react against a genetically incompatible graft producing
a host-versus-graft response, an immunocompetent graft (such as bone
marrow or intestinal tissue) may react against the host resulting in
graft-versus-host disease. These reactions are mediated by allogenic
responses directed against, a foreign MHC molecule and are mimicked in
vitro by the mixed lymphocyte reaction (MLR). Graft/host interactions
result in chronic inflammation surrounding the grafted tissue with an
increase in markers of immune activation such as are seen in AIDS (Grant,
Immunol. Today 12 171-253. (1991)). Treatment of the graft/host
interactions currently include either azathoprine, cyclosporin A or
methylpredisone and more recently, rapamycin (Stepkowski et al,
Transplantation 53 254-264 (1992); Huber et al, Bibliotheca Cardiologica.
43 103-110 (1988)). Monoclonal antibodies specific for CD3 (Wissing et al,
Clin. Exp. Immunol. 83 333-337 (1991)) and CD4 (Reinke et al, Lancet 338
702-703 (1991)) have been used experimentally to inhibit graft/host
The present invention deals with the identification of a group of peptides
that alleviates the inflammatory response in a number of diseases. These
include: autoimmune disease, organ transplantation; neoplasia; viral,
bacterial, fungal or other infections; and, in particular, any disease
wherein infection can manifest in an opportunistic fashion, eg during
cytotoxic or radiation therapy or in any situation where an
immunodepressed state exists. Peptides useful in the present invention do
not necessarily interfere directly with the pathogenic mechanisms of the
disease-causing component. As will be described below in the experimental
data, the mechanism whereby these peptides can alleviate the symptoms of
these diseases is dependent on their capability of modulating the
production and effect of cytokines produced by activated cells of the
immune system. The modulation of cytokines may not be limited to TNF but
may also be valid for a whole range of interleukins, for example from
interleukin-1 to interleukin-10. The data presented are at present not
direct evidence, but rather a powerful indirect model. Thus, the model
uses one of the most powerful inflammatory compounds known, LPC, which
binds to receptors on neutrophils, monocytes and macrophages; these cells
consequently become activated and start production of IL-1 and TNF, among
other cytokines, thus starting the inflammatory cascade. One parameter
used to measure this effect of LPS is the concentration of blood glucose,
which normally decreased on exposure to TNF or LPS. From what is known in
the literature about the mechanism of Peptide T at a cellular level, it is
therefore highly surprising that Peptide T and its analogues are able
significantly to reduce the negative effects of LPS. LPS normally combines
with LPS-Binding Protein (LBP) and exerts its dramatic effect through the
CD14 receptor. In the literature up to date, the peptides useful in this
invention have only been connected to the CD4 receptor, which is not
believed to be involved in the primary inflammatory response associated
with cytokines, such as TNF, or LPS.
More specifically, it has been discovered that a particular group of
peptides, particularly those within the group having at least 5 amino acid
residues, are very effective agents useful in the treatment, inter alis,
MS and HAM, and are likely to be useful in treating other myelopathies,
most of which have similar disease mechanisms.
From the above discussion, it is apparent that many symptoms and diseases
are associated with chronic inflammation; however, several of these
diseases appear to involve different mechanisms. It is therefore important
that particular compounds have been found which are useful in treating
symptoms and diseases associated with chronic inflammation where it
appears that these compounds interact in some manner with CD4 receptors of
immune system cells. The compounds relate, as indicated above, to Peptide
T and its various derivatives. It was originally thought that such
compounds had no effect on the immune system other than being very useful
in blocking attachment of HIV virus to CD4 receptor cells (Ruff et al, IV
Interantional Conference on AIDS, Stockholm June 1988).
Originally, many of the peptides useful in the invention were described as
being effective in the prevention of infection and replication of HIV in
vitro; see EP-A-0249390, EP-A-0249394 and WO-A-8809338, all of which are
incorporated by reference to the maximum extent allowed by law, as are all
other documents referred to in this specification. All compounds disclosed
in these specifications are useful for the present invention. The original
peptide has its basic point of origin in the octapeptide Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Tyr.
It is called Peptide T because 50% of the amino acid residues are
threonines. This peptide has been identified from a subregion of the human
immune deficiency virus (HIV) external glycoprotein molecule gp120, which
is responsible for binding to any cell carrying the CD4 molecule and, in
particular, helper lymphocytes, microglial cells in the CNS, monocytes and
dendritic cells. Binding occurs via specific attachment of gp120 the CD4
molecule. Treating individuals infected with HIV with this peptide and its
derivatives, which are described below, consequently has the effect of
potentially inhibiting binding of the whole virus or the neutotoxic gp120
molecule to the cell receptor CD4. In this way, the cell is protected from
infection, and so the virus, being unable to replicate, is destroyed by
the immune defence.
According to a first aspect of the present invention, there is provided
the use of a linear or cyclic peptide of General Formula 1:
I-A-B-C-D-E-F-G-H-II General Formula 1) (SEQ. ID. NO: 1))
wherein A is Ala, Gly, Val, Ser, Thr or absent,
B is Ala, Gly, Val, Ser. Thr or absent,
C is Sef, Thr or absent,
D is Ser, Thr, Asn, Glu, Arg, Ile, Leu or absent,
E is Ser, Thz, Asp or absent,
F is Thr, Ser, Asn, Arg, Gln, Lys, Trp or absent,
G is Tyr or absent,
H is Thr, Arg, Gly, Met, Met (O), Cys, Thr, Gly or absent,
I is Cys or absent,
II is Cys or absent,
at least one of the amino acids optionally being substituted by a
monomeric or polymeric carbohydrate or derivative thereof, such
substitution being accomplished through hydroxyl and/or amino and/or amido
groups of the amino acids.
and wherein the peptide comprises at least four amino acid residues,
or a pharmaceutically acceptable salt thereof, in the manufacture of a
medicament for treating or preventing inflammation.
Each of the amino acids referred to in General Formula 1 may be in the L-
or D- stereoisomeric configuration and candidates for H may be esterified
or amidated. The peptide comprises at least 4 amino acids.
Tetra-, penta-, hexa-, hepta-, octa- and nona-peptides useful in the
invention are all of the peptides chosen from the sequence:
by deleting residues, for example, one at a time, from either the carboxyl
or amino terminal, or from within the sequence.
It is appreciated that peptides having the core sequence of
Thr-Thr-Asn-Tyr-Thr- may have at both ends additional amino acid residues,
some of which are represented by General Formula 2:
X-Ser-Thr-Thr-Thr-Asn-Tyr-Y (General Formula 2)(SEQ ID NO:2)
wherein X is an amino acid terminal residue selected from Ala and D-Ala
and Y is a carboxy terminal residue selected from Thr and Thr-amide.
A particular preferred peptide of the group of peptides has the
aforementioned core sequence of -Thr-Thr-Asn-Tyr-Thr -. These peptides of
the above General Formula 2, and in particular a variant Peptide T of the
formula -Ser-Thr-Thr-Thr-Asn-Tyr-, were found to be very useful in
inhibiting binding of the human immunodeficiency virus (HIV) to human
cells by blocking receptor sites on the cell surfaces. The term Peptide T
is used throughout the specification to reference, unless the context
otherwise requires, peptides of General Formula 2 which all include the
core peptide sequence. It is therefore intended that Peptide T encompass
all of the compounds of General Formula 2 where it is understood that all
such compounds are variants of the normally understood octapeptide T, also
referred to as prototype Peptide T, of the particular formula S-Ala-Ser-Thr-Thr-The-Asn-Tyr-Thr-amide
(SEQ ID NO:3).
The invention may be useful in both clinical (human) and veterinary
medicine. The invention therefore has application in a method for treating
or preventing inflammation, the method comprising administering to a human
or other animal subject, for example on a repeated basis, a peptide of
General Formula 1. The peptide will generally be administered in an
effective, non-toxic amount or in such an amount that strikes an
acceptable balance between efficacy and toxicity, having regard to the
circumstances of the case.
Preferred peptides useful in the invention have, as their active portion,
an amino acid sequence of the formula:
-Thr-Thr-Asn-Tyr-Thr-(SEQ ID NO: 4).
These peptides, while being useful for all prophylactic and therapeutic
utilities within the invention, are particularly preferred for the
prevention or treatment of MS and HAM and for the prevention or treatment
of symptoms or diseases, in humans or other animals, associated with
chronic immune activation, chronic inflammation and chronic autoimmune
Most preferred peptides useful in the invention, then, are the following:
1. D-Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr-NH2 (prototype Peptide T)
(SEQ ID NO: 3)
2. Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr (SEQ ID NO: 5)
3. D-Ala-Ser-Thr-Thr-Thr-Asn-Tyr-Thr (SEQ ID NO: 3)
4. D-Ala-Ala-Ser-Ser-Ser-Asn-Tyr-Met (SEQ ID NO: 6)
5. Thr-Asp-Asn-Tyr-Thr (SEQ ID NO: 7)
6. Thr-Thr-Ser-Tyr-Thr (SEQ ID NO: 8)
7. Thr-Thr-Asn-Tyr-Thr (SEQ ID NO: 4)
8. D-Thr-Thr-Tyr-D-Thr (SEQ ID NO: 9)
9. D-Ala-Ser-D-Thr-Thr-D-Thr-Asn-Tyr-D-Thr-NH2 (SEQ ID NO: 10)
10. D-Ser-Ser-D-Thr-Thr-D-Thr-Thr-Tyr-D-Thr-NH2 (SEQ ID NO: 11)
Quite often it may be an advantage to have the amino terminal amino acid
as a D-stereoisomer, to protect the molecule from degradation from
aminopeptidases; alternatively or additionally, the carboxy terminal amino
acid may be an amino acid amide to protect the molecule from degradation
from carboxypeptidases. In this connection, compounds 5, 6 and 7, listed
above, include analogues with D-Thr as the amino terminal residue and/or
an amide derivative at the carboxy terminal.
Furthermore, it should be understood that one more of the amino acids in
the peptides may be substituted N-alkyl (e.g. (C1 -C4)
alkyl) amino acids instead of primary amino acids; examples include methyl
and ethyl. The hydroxyl group side chains of one or more of the amino
acids (Ser, Thr, Tyr) may be derivatised into an ether or ester group. Any
(optionally substituted) alkyl ester or ether may be formed, such as (C1
-C4) alkyl, aryl or aryl (C1 -C4) alkyl
esters, ethers, thioesters and thioethers, for example phenylester,
benzylether or thiophenol ethylester. The presently preferred ethers are
methyl, ethyl and propyl ethers and presently preferred esters are methyl,
ethyl and propyl esters.
The hydroxyl side chains of the amino acids Ser, Thr and/or Tyr and the
amido groups of the amino acids Asn and/or Gln may be substituted with
different carbohydrates or derivatives of carbohydrates. Carbohydrate
derivatives may be as discussed above.
Linear peptides useful in this invention may be prepared by any suitable
process, such as conventional solid phase peptide synthetic techniques;
see "Solid Phase Peptide Synthetic Techniques", 2nd ed, J. M.
Stewart, J. D. Young.
Pierce Chemical Company, 1984, ISBN: 0-935940-03-0. A frequently used
solid phase method is the Merrifield technique. Another possibility is
solution phase techniques. The preferred peptide, prototype Peptide T, is
readily obtainable from Carlbiotech A/S, Copenhagen, Denmark.
Cyclic peptides useful in the invention may be prepared by known
techniques, such as, for example, described in Y. Hamada in Tetrahedron
Letters, 26 5155 (1985). Cyclic peptides may be established in the form of
a disulphide bridge between two Cys residues and/or by reacting the
carboxy terminal amino acid residue with the amino terminal residue and/or
by reacting the amino terminal residue with for example the .gamma.-
carboxyl group of Glu, when Glu is at position D.
Carbohydrate derivatives may be prepared by methods known in the art.
Certain peptide derivatives useful in the invention are new and themselves
form another aspect of the invention according to which there is provided
a linear or cyclic peptide of General Formula 1:
I-A-B-C-D-E-F-G-H-II (General Formula 1)
wherein A is Ala, Gly, Val, Ser, Thr or absent,
B is Ala, Gly, Val, Ser, Thr or absent,
C is Ser, Thr or absent,
D is Ser, Thr, Asn, Glu, Arg, Arg, Ile, Leu or absent,
E is Ser, Thr, Asp or absent,
F is Thr, Ser, Asn, Arg, Gln, Lys, Trp or absent,
G is Tyr or absent,
H is Thr, Arg, Gly, Met, Met(O), Cys, Thr, Gly or
I is Cys or absent,
II is Cys or absent,
at least one of the amino acids being substituted by a monomeric or
polymeric carbohydrate or derivative thereof, such substitution being
accomplished through hydroxyl and/or amino and/or amido groups of the
and wherein the peptide comprises at least four amino acid residues,
except for glycosylated prototype Peptide T,
or a pharmaceutically acceptable salt thereof.
Glycosylated Peptide T is disclosed in Urge et al., Biochem. Biophys. Res.
Comms. 184(2) 1125-1132 (1992), published Apr. 30, 1992, but the utility
of the present invention is neither disclosed nor suggested.
Preferred features of this aspect of the invention are as for the first
Peptides useful in the invention may be administered as a composition in
conjunction with a pharmaceutically acceptable carrier.
In this way the peptides can be used in pharmaceutical compositions and
compositions of matter for treating and preventing any disease or
condition caused by an organism, compound or immune dysfunction that
results in an inflammatory reaction of the immune system.
The peptides or peptide formulations may be used alone or in combination
with any other pharmaceutically active compound, such as an anti-infective
agent, for example an antibiotic and/or antiviral agent and/or antifungal
agent, or another pharmaceutically active compound, such as an anti-neoplastic
The peptides may be administered orally, bucally, parenterally, topically,
rectally, vaginally, by intranasal inhalation spray, by intrapulmonary
inhalation or in other ways.
In particular, the peptides according to the invention may be formulated
for topical use, for inhalation with spray or powder, for injection (for
example subcutaneous, intramuscular, intravenous, intra-articular or
intra-cisternal injection), for infusion or for oral administration and
may be presented in unit dose form in ampoules or tablets or in multidose
vials or other containers with an added perservative. The compositions may
take such forms as suspensions, solutions, or emulsions or gels in oily or
aqueous vehicles, and may contain formulatory agents such as suspending,
stabilising and/or dispersing agents. Alternatively, the active ingredient
may be in powder and/or lyophilised form for direct administration or for
constitution with a suitable vehicle (e.g. sterile, pyrogen-free water,
normal saline or 5% dextrose) before use. The pharmaceutical compositions
containing peptides(s) may also contain other active ingredients such as
antimicrobial agents, or preservatives.
The compositions may contain from 0.001-99% (w/v or, preferably, w/w) of
the active material. Peptide T obtainable from Carlbiotech A/S is usually
formulated and packaged in a sterile manner in 5% dextrose solution in
multi-dose vials. It will be appreciated that the peptide may be packaged
in other carriers, such as saline. Preferably, the concentration of
peptide in each dose is in the order of 8.5 mg/ml for subcutaneous
injection in one ml doses.
The compositions are administered in therapeutically or prophylactic
effective does, i.e. 0.05-10000 mg of peptide per day, in particular
5-1000 mg per day. Very large doses may be used as the peptide according
to the invention is non-toxic. However, normally this is not required. The
dose administered daily of course depends on the degree of inflammation
and inflammatory response.
For administration by injection or infusion of the compositions, the daily
dosage, as employed for treatment of adults of approximately 70 kg of body
weight, will often range from 0.2 mg to 20 mg of active material which may
be administered in the form of 1 to 4 doses over each day, such dosage
ranges depending upon the route of administration and the condition of the
Compositions as described above may be prepared by mixing or otherwise
bringing into association the ingredients.
The invention may be useful in the prevention or treatment of illness or
medical conditions, particularly those involving inflammation, such as:
Viral, bacterial or drug-induced hepatitis or meningitis;
Rheumatoid, psoriatic, reactive, or osteo-arthritisor other arthritides;
ARDS (adult respiratory distress syndrome);
Inflammation secondary to the chemotherapy or radiotherapy of neoplastic
The invention finds particular use in the prevention or treatment of MS,
HAM and other inflammatory myelopathies (particularly those previously
specifically mentioned) and/or symptoms or diseases in humans which are
associated with chronic immune activation. More particularly, the
invention is useful in treating chronic fatigue syndrome, toxic shock
syndrome associated with Staphylococcus aureus infection, arthritis,
inflammatory bowel disease and host-versus-graft response in transplant
patients. Such efficacious results in the use of the above compounds is
thought to be due, without being limited to any particular theory, to the
immunosuppressive activities of these compounds in chronic inflammatory
In order to provide a guideline for the administration of and insight into
the use of the peptides according to the invention, particularly the
treatment of MS, myelopathies such as HAM and chronic inflammation, and
the formulation of the compositions, the following is offered as a guide
based on the extensive work already conducted in the use of peptide T for
treating HIV infection.
Peptide T is an octapeptide homologous to a region of gp120, an HIV
envelope glycoprotein, and to human vasoactive intestinal peptide (VIP).
It was originally developed by Pert et al (EP-A-0249394), to block the
binding of gp120 (an HIV envelope glycoprotein) and thus also block
binding of HIV to CD4, the specific membrane-bound vial receptor, thereby
blocking internalisation of the virus into the cell--a process necessary
for viral replication. The CD4 molecule necessary for the entry of HIV
into cells has been localised on the surface of lymphocytes, macrophages,
microglial cells, neurons and numerous other cells. Binding of HIV to the
CD4 receptor has been demonstrated to effect viral entry; and binding of
free (non-viral related) gp120 has resulted in neuronal toxicity in both
in vitro and in vivo studies.
The efficacy of Peptide T in reversing signs of HIV-induced dementia has
been demonstrated in both the Peptide T Phase I clinical trial at the
University of Southern California in Los Angeles and in the Phase II
clinical trial at the Fenway Clinic in Boston. Both studies have
demonstrated improvement in the HIV-induced neurocognitive impairment in
patients with AIDS.
To date, the Peptide T/gp120/VIP homology has been used to explain at
least two possible mechanisms of action of Peptide T. Firstly, that it
competitively binds to CD4 (the known receptor for HIV) on human cell
surfaces and competes with both HIV and gp120 for binding sites.
The binding of Peptide T and its analogues of General Formula 1, or more
particularly General Formula 2, to CD4 could produce a blocking effect to
prevent the binding of any other molecule capable of binding to that
receptor; alternatively, or in addition, the binding of Peptide T to CD4
could induce a reaction similar to that caused by the endogeneous ligand.
CD4 is the differentiation antigen that defines the T lymphocyte subgroup
of helper/inducer cells, but it is also present on a wide variety of cells
including neurons, activated macrophages and B cells. CD4 is the
predominant receptor for HIV and was originally thought to be necessary
for cellular infection. Using the monoclonal antibody OKT4, Pert et al
(Proc. Natl. Sci. U.S.A. 83 9254-9258 (1986); Pert et al (Clin.
Neuropharmacol. 9(4) S198 (1986)) demonstrated the presence of this
antigen throughout the human CNS and showed that it is present in highest
concentration in the dentate gyrus, hippocampus, amygdala and deep cortex.
This distribution was found to be similar to that found in other higher
mammals. Peptide T and similar analogues were found to inhibit and binding
of radiolabelled gp120 to rat hippocampal membranes and to do so in 0.1 nM
Using Peptide T and the same analogues, Pert et al (Proc. Natl. Sci.
U.S.A. 83 9254-9258 (1986); Pert et al (Clin. Neuropharmacol. 9(4) S198
(1986)) were able to demonstrate a reduction in the detectable levels of
HIV reverse transcriptase when these peptides were present in an assay of
HIV infectivity. A ninefold reduction of reverse transcriptase took place
at 100 nM concentrations of Peptide T.
Since gp120 is not identical in all isolated strains of HIV, a comparison
was made with nine different HIV isolates Pert et al (Clin. Neuropharmacol.
9(4) S198 (1986)). Significant homology was found between the isolates
examined and Peptide T when comparison was made with the core pentapeptide.
(Ruff et al. FEBS Lett. 211 17-22 (1987); Brenneman et al. Nature, 335
639-642 (1988); Brenneman et al, Drug Dev. Res. 15 361-369 (1988);
Komisaruk et al, Annals of the NY Acad. Sci. 527 650-654 (1988); Buzy et
al. The Lancet 22 Jul., 226-227 (1989)). This comparison has now been
extended to over twenty isolates.
The inhibition of gp120 and HIV binding to CD4, as well as the
demonstration of reduced infectivity of HIV in the presence of Peptide T
and its analogues, provides one possible mechanism of action to explain
the clinical effects of Peptide T. In this regard, Peptide T in sufficient
concentration may prevent new cellular infection with HIV. Initial
research in this area was focused on the CNS for two reasons: a high
concentration of CD4 molecules was found on neurons and one of the major
effects of HIV infection is the development of neurocognitive dysfunction.
These fats are particularly important given that Peptide T is transported
from the blood to the brain by an active, saturable transport system,
while its exit is by diffusion only, (Barrera et al, Brain Res. Bull. 19
Although it is well accepted that HIV can infect not only lymphocytes but
also neurons, it is difficult to ascribe the neurologic dysfunction seen
in HIV patients to active CNS HIV infection, since only a small number of
neurons are actively infected. It has been suggested that the neurological
deficits seen in HIV injection may occur not only as a result of infection
but also as a result of a viral "toxin", such as gp120.
Brenneman et al, Nature 335 639-642 (1988)) found that purified gp120 from
two isolates as well as recombinant gp120 produced significant neuronal
cell death in cultures of mouse foetal hippocampal neurons. Neurotoxicity
could be reduced by pretreatment with antibody to CD4 and was completely
eliminated by VIP. Since mouse neurons cannot be infected with HIV, it is
evident that neurotoxicity is gp120-induced and is not a result of viral
entry or replication.
VIP (66, 67, 68, 69: TDNYT) and the core peptide (61-65: TTNYT) share the
homologous sequence that binds CD4, and that is also found in isolates of
the much larger gp120. Peptide T, when used in the same mouse hippocampal
neuronal culture system, completely, antigonised the gp120-induced
neurotoxicity, Brenneman et al, Drug Dev. Res. 15 361-369 (1988)). In
addition, CSF from a patient with AIDS dementia produced substantial
neurotoxicity in this system (4-49%) killing at 1:100,000 dilution). This
effect was inhibited by Peptide T, Buzy et al, The Lancet, July 22, pp
226-227, (1989)). Normally gp120 is produced in vast excess of amounts
required for viral replication; this excess gp120 may exert a neurotoxic
effect far out of proportion to the number of microglial cells or neurons
actively infected with HIV.
Peptide T may act as an agonist in addition to or even in the absence of
its neuroprotective effects against viral infection and neurotoxicity.
Direct agonist activity has been demonstrated in two ways. Ruff et al, (FEBS
Lett. 211 17-22 (1987)) showed that peptide T and two analogues were
potent agonists of human monocyte chemotaxis. Their rank order potency as
chemotactic agents corresponded to their relative ability to inhibit both
gp120 binding and HIV T cell infectivity.
As a further demonstration of the agonist activity of Peptide T, both
Peptide T and VIP exert their cellular effects via the regulation of
protein kinase C: Zorn et al, The Endoc. Society. Abstract (1989)).
Agonist activity of Peptide T is thus implied by the production of a
transmembrane signal that can influence the regulation of protein kinase
C. It has also been demonstrated in six individual experiments, as part of
the present invention, that Peptide T can down-regulate the enzyme p56lck,
a tyrosine kinase linked to the cytoplasmic portion of membrane-bound CD4,
thus implying that the binding of Peptide T to the CD4 molecule can
produce a transmembrane signal.
Further evidence of Peptide T's potential VIP-like agonist activity is
provided by results from experimental testing of the hypothesis of
Komisaruk et al, Annals of the NY Acad. Sci. 527 650-654 (1988) that VIP
released from pelvic nerve terminals into the spinal cord can produce
analgesia. Knowing that naloxone-independent analgesia produced by
administration of VIP to the periaqueductal grey matter in rats had been
shown, the investigators administered VIP directly to rat spinal cord and
measured the pain threshold to distal noxious stimuli to test the
hypothesis. Spinal administration of VIP produced analgesia as measured by
the tail-flick latency response and the tail-shock induced vocalisation
test by action on both opiate and non-opiate modulated pain pathways,
Komisaruk et al, Annals of the NY Acad. Sci. 527 650-654 (1988)).
The existence of clinical benefits from the administration of Peptide T to
humans has been suggested in all studies to data: in HIV disease, by the
Pilot Swedish Study, the USC Phase I and Fenway/CRI studies, and the
Toronto Western Hospital Compassionate Use Program involving 51 patients;
in psoriasis and other medical conditions, in case reports from Sweden (Marcusson,
Lazega et al, 1989-9, and Marcusson and Wetterberg, 189-10) and in 8
patients with psoriasis or other medical conditions in Toronto.
Neuroncognitive improvement found in HIV positive patients and improvement
in constitutional symptoms in both HIV positive and HIV negative patients
may well depend primarily on Peptide T's VIP-like neurotropic and agonist
effects, as well as the anti-inflammatory and anti-TNF effects discussed
Not wishing to be bound by any particular theory, with respect to the use
of these peptides with treatment of MS and HAM, and in view of the above
guidelines and discussions in relation to the use of various peptides of
General Formula 2 and their analgoues in the treatment of HIV, it is
hypothesised that there are numerous similarities of disease expression
and potential similarities of disease etiology. Peptide T appears to act
as an agonist and as a blocker of CD4-mediated immune function rather than
as an antiviral drug. In investigations associated with the present
invention, patients with non-HIV disease such as multiple sclerosis,
HTLV-1 associated myelopathy, and psoriasis have all been treated with
Now that the effectiveness of the peptides of General Formula 1, and
particularly those of General Formula 2, is shown, the following is
suggested as a hypothesis as to why the compounds do work:
1) Both HAM and MS are chronic CNS inflammatory and demyelinating diseases
as is HIV disease.
2) Both diseases have possible viral aetiologies; it is now generally
accepted that HAM is caused by the retrovirus, HTLV-1, a virus in the same
family as HIV; MS has also been suggested as a manifestation of HTLV-1
infection and the chronic fatigue syndrome has recently been linked to a
number of possible viral infections both of DNA (e.g. HHV-6) and
3) The two diseases share a number of common symptoms, for example,
fatigue, lack of balance and signs of autoimmune phenomena; it is worth
while noting that HTLV-1 disease exhibits numerous signs of autoimmunity
such that it may be expected that some retroviral diseases have a
concomitant expression in autoimmune phenomena. One common theme among
these diseases may be peripheral neuropathy which is based on the process
4) The basis appears to be the common denominator of both demyelination
whether it be in the central or peripheral nervous system and the common
autoimmune manifestations in HAM, MS and HIV disease.
5) Demyelination is associated with inflammation of any aetiology and
appears to be mediated at least in part by TNF.
Not wishing to be bound by any particular theory with respect to the use
of peptides with the treatment of symptoms and diseases associated with
chronic immune activation, and in view of the above guidelines and
discussions in relation to the use of various peptides of General Formula
2 and their analogues in the treatment of HIV, it is hypothesised that
there is an immunomodulatory effect of the Peptide T binding to CD4. Such
effect is demonstrated in the following examples where Peptide T has been
found to inhibit mitogen induced proliferation of peripheral blood
mononuclear cells (PBMC) at picomolar and lower concentrations. We have
found that Peptide T allowed PBMC to proliferate in response to mitogen,
but at a reduced level compared to the growth of PMBC in its absence.
Pre-incubation of PBMC with Peptide T for less than 30 minutes had no
effect on mitogen stimulation. However, exposure of PBMC to Peptide T for
2 hours followed by washing of the cells before exposure to the mitogen
resulted in inhibition of proliferation similar to that seen when cells
are incubated in the presence of both Peptide T and mitogen. It was also
found that Peptide T did not significantly affect the growth of PBMC
cultured in the absence of mitogen. It is therefore thought that Peptide T
is able to suppress the normal proliferative response of PBMC to non-CD4
associated proliferation signals.
Now that the effectiveness of the peptides of General Formula 1, and
particularly those of General Formula 2, in treating symptoms and diseases
associated with chronic immune activations or chromic inflammation has
been discovered, the following is suggested as hypothesis of the mechanism
of action of Peptide T and, therefore, why the compounds useful in the
invention are effective.
Peptide T binds to CD4. It has been established in the following tests
that peptide T inhibits mitogen and MLR induced lymphocyte proliferation.
It is therefore thought that Peptide T may serve as an immunomodulatory
drug which would down-regulate the enhanced immune response occurring in
the chronic presence of antigen or for other reasons mentioned and hence
reduce chronic inflammation.
Underlying all these utilities runs the common theme of inflammation.
In accordance with various embodiments of this invention and in view of
the above guidelines gained from the use of peptides of General Formula 1,
and particularly those of General Formula 2, in the treatment of HIV,
similar doses of Peptide T and its analogues can be administered to humans
or other animals for purposes of treating inflammation.
Claim 1 of 17 Claims
What is claimed is:
1. A method for treating or preventing inflammation in a subject by
administering an effective amount of the peptide:
I-A-B-C-D-E-F-G-H-II (General Formula I)
A is Ala, Gly, Val, Ser, Thr or absent,
B is Ala, Gly, Val, Ser, Thr or absent,
C is Ser, Thr or absent,
D is Ser, Thr Asn, Glu, SArg, Ile, Leu or absent,
E is Ser, Thr, Asp or absent,
F is Thr, Ser, Asn, Arg, Gln, Lys, Trp or absent,
G is Tyr or absent,
H is Thr, Arg, Gly, Met, Met(O), Gys, Thr, Gly or absent,
I is Cys or absent,
II is Cys, an amide group, an ester group or absent,
at least one of the amino acids optionally being substituted by a
monomeric or polymeric carbohydrate or an alkyl ester or alkyl ether
derivative thereof, such substitute being accomplished through hydroxyl
and/or amino groups of the amino acids, and wherein the peptide comprises
at least 4 amino acid residues, or a pharmaceutically acceptable salt
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