WO2006101910A2

WO2006101910A2 - Peptide compounds and methods of using peptide compounds to treat conditions affecting the cardiac, pulmonary, and nervous systems

Info

Publication number: WO2006101910A2
Application number: PCT/US2006/009363
Authority: WO
Inventors: Steven Kates
Original assignee: Ischemix, Inc.
Priority date: 2005-03-16
Filing date: 2006-03-14
Publication date: 2006-09-28
Also published as: WO2006101910A3

Abstract

The present invention provides a method of treating or preventing diseases or conditions affecting the cardiac, pulmonary and/or nervous systems in a subject using peptide compounds. The methods of the invention are useful for the treatment and prevention of a variety of disorders or conditions including, without limitation, ischemia, stroke, pulmonary diseases, allergic reactions, physical trauma to pulmonary tissues, exposure to harmful entities in the environment, and smoking.

Description

PEPTIDE COMPOUNDS AND METHODS OF USING PEPTIDE COMPOUNDS TO TREAT CONDITIONS AFFECTING THE CARDIAC PULMONARY AND NERVOUS

SYSTEMS

FIELD OF THE INVENTION

[0001] This invention relates to the therapeutic and prophylactic use of peptide compounds for treating or preventing diseases affecting the cardiac, pulmonary and nervous systems.

BACKGROUND OF THE INVENTION

[0002] Pulmonary inflammation includes the inflammation of the trachea and bronchial air passage and other areas of the respiratory system. Pulmonary inflammation occurs in a variety of disorders and conditions including, without limitation, pulmonary diseases, such as asthma, cystic fibrosis, emphysema, bronchitis, obstructive pulmonary disease, bronchitasis, physical trauma to pulmonary tissues, infections of the pulmonary system, and smoking. [0003] Asthma is a disease of the respiratory airway characterized by hyper responsiveness of the tracheobronchial tree to a multiplicity of stimuli. Pulmonary inflammation is typically associated with asthma and is a primary contributor to the symptoms of asthma. The symptoms of asthma include shortness of breath, coughing and wheezing. Asthma affects about 4 to 5 percent of the population of the United States, and occurs at all ages, but predominantly in early life.

[0004] Cystic Fibrosis (CF) is a multisystem disease that leads to dysfunction of many organs including the lungs, pancreas, sweat glands, and urogenital system. Respiratory tract disease is almost universal in CF patients. Over the course of years lung function in CF patients deteriorates and may eventually lead to respiratory failure. Repeated lung infections and buildup of secretions in CF patients are accompanied by pulmonary inflammation. More than 95 percent of CF patients die of complications resulting from lung infection.

[0005] The prevalence of CF varies among ethnic groups. CF is detected in approximately 1 in 2500 live births in the Caucasian population of North America and northern

Europe, 1 in 17,000 live births of African- Americans, and 1 in 90,000 live births of the Asian population of Hawaii.

[0006] Chronic bronchitis is a condition associated with excessive tracheobronchial mucus production. Emphysema is a lung condition featuring an abnormal accumulation of air in the lung's many tiny air sacs called alveoli. As air continues to collect in these sacs, they become enlarged, and may break, or be damaged and form scar tissue. Emphysema is strongly associated with smoking cigarettes. It can also be associated with or worsened by repeated infection of the lungs. Chronic obstructive pulmonary disease is a condition in which there is a chronic obstruction of airflow due to chronic bronchitis and/or emphysema.

[0007] Approximately 20 percent of adult males have chronic bronchitis.

Approximately two-thirds of adult males and one-fourth of females develop emphysema in varying degrees of severity.

[0008] Bronchiectasis is an abnormal and permanent dilatation of bronchi. Symptoms of bronchiectasis include persistent or recurrent cough and thick mucus production. Bronchiectasis is a consequence of inflammation and destruction of the structural components of the bronchial wall. A single or recurrent respiratory infection triggers bronchial wall inflammation and destruction in most cases.

[0009] The peptide compounds of the present invention are also useful for treating or preventing diseases and conditions, such as cognitive degeneration, senility, Tardive dyskinesia, head trauma, brain and/or spinal cord trauma, oxygen toxicity, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer's disease, diabetes, ulcerative colitis, leukemia and other cancers characterized by elevation of ROS or free radicals, Downs syndrome, macular degeneration, cataracts, schizophrenia, epilepsy, septic shock, polytraumatous shock, burn injuries, and radiation-induced elevation of ROS.

[0010] The peptide compounds of the present invention are also useful for treating or preventing ischemia, including different subtypes of ischemia, including, but not limited to, cerebral ischemia, cerebral infarction, intracranial hemorrhage (collectively known as "ischemic cerebrovascular disease"), anoxic-ischemic encephalopathy, ischemic heart disease, ischemic renal disease, intestinal ischemia (including mesenteric ischemia and infarction, ischemic colitis, and angiodysplasia), and myocardial ischemia (including obstructive DAD, myocardial infarction and myocardial ischemia). Thus, the compositions of the present invention are useful for treating or preventing different subtypes of ischemia.

[0011] One clinical manifestation of cerebral ischemia is stroke, which could be due to either ischemic cerebral infarction or brain hemorrhage. The compositions of the present invention are particularly suitable for treating or preventing stroke in a subject, preferably in a human, in need thereof.

[0012] The aforementioned diseases are just a few examples of diseases which may be treated or prevented by using the compounds, compositions and methods described herein. Such diseases are associated with a large number of disorders and conditions, some of which are severe and sometimes life-threatening. The present invention provides novel methods of treating or preventing the diseases described above using the peptide compounds described herein.

SUMMARY OF THE INVENTION

[0013] The present invention is based on the surprising discovery that certain peptide compounds are able to inhibit and/or diminish the onset of diseases affecting the cardiac, pulmonary and nervous systems, and includes methods of treating or preventing diseases affecting the cardiac, pulmonary and nervous systems. In particular, the present invention includes a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to a subject an effective amount of at least one of the peptide compounds described in the present invention. [0014] In one embodiment, the present invention describes a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof by administering to the subject an effective amount of at least one peptide compound, wherein the peptide compound contains fewer than 20, and, in order of increasing preference, fewer than about 18, 15, 13, 9, 8, 7, 6, 5, 4 and 3 amino acids in length.

[0015] In another embodiment, the invention provides a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to a subject an effective amount of at least one peptide compound that treats or prevents the diseases described herein at low concentrations.

[0016] Pulmonary inflammation is associated with various diseases and conditions, including, without limitation, asthma, cystic fibrosis, chronic bronchitis, chronic obstructive pulmonary disease, emphysema and brochiectasis. Thus, another embodiment of the present invention is a method of treating or preventing a disease in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound described herein, wherein the disease or condition is selected from a group consisting of asthma, cystic fibrosis, chronic obstructive bronchitis, chronic bronchitis, broncholithiasis, chronic obstructive pulmonary disease, chronic granulomatous lung disease, eosinophilic pneumonias, hypersensitivity pneumonitis, idiopathic pulmonary fibrosis, pneumonia, emphysema and bronchiectasis (hereinafter, collectively known as "pulmonary diseases"). [0017] This combination therapy is also useful for treating or preventing diseases and conditions, such as cognitive degeneration, senility, Tardive dyskinesia, head trauma, brain US2006/009363

and/or spinal cord trauma, oxygen toxicity, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer's disease, diabetes, ulcerative colitis, leukemia and other cancers characterized by elevation of reactive oxygen species ("ROS") or free radicals,

Downs syndrome, macular degeneration, cataracts, schizophrenia, epilepsy, septic shock, polytraumatous shock, burn injuries, and radiation-induced elevation of ROS (hereinafter, collectively known as "diseases affecting the nervous system").

[0018] It is particularly useful for treating or preventing ischemia, including different subtypes of ischemia, such as cerebral ischemia (also known as "ischemic cerebrovascular disease"), anoxic-ischemic encephalopathy, ischemic heart disease, intestinal ischemia, ischemia relating to renal disease or injury, and myocardial ischemia (also known as "myocardical infarction") (hereinafter, collectively known as "ischemic diseases").

[0019] The present invention also includes methods of preparing and administering pharmaceutical compositions useful for treating or preventing the diseases described in detail above.

[0020] Additional aspects of the present invention will be apparent in view of the description which follows.

BRIEF DESCRIPTION OF THE FIGURES

[0021] Figure 1 shows diagrams of structures of representative thyronine groups

(thyronyl) that may serve as amino terminal capping groups of peptide compounds used in the invention. Each structure shown in Figure 1 is a form of a thyronine group that may be linked at its carbonyl carbon to the amino terminal amino group of a peptide. Some internal numbers (3, 3', 5, 5') are shown to indicate positions of iodine substitution in one or both phenyl groups of the thyronine group. "Thyroxine T₄" refers to the diagrammed tetraiodothyronine group, which is substituted with iodine at phenyl positions 3, 5, 3', and 5', and which is derived from the thyroxine hormone. "3, 5, 3' T₃" refers to the diagrammed triiodothyronine group, which is substituted with iodine at phenyl positions 3, 5, and 3'. "3, 3', 5' T₃" refers to the diagrammed triiodothyronine group, which is substituted with iodine at phenyl positions 3, 3', and 5'. "T₀" refers to the diagrammed thyronine group, which has no iodine substitutions.

[0022] Figure 2 shows an increase of inflammatory response in rats when exposed to different concentrations of urban dusts (PMlO).

[0023] Figure 3 shows that CMX-11 suppresses inflammatory response induced by urban dusts (PMlO) in rats.

DETAILED DESCRIPTION OF THE INVENTION

[0024] in order that the invention may be better understood, the following terms are defined.

[0025] "Peptide compound" as understood and used herein includes unmodified or modified, derivatized or underivatized peptides. Such peptide compounds typically contain fewer than 20 amino acids. Derivative or derivatized peptides contain one or more chemical moieties (other than amino acids) that are covalently attached at the amino terminal amino acid residue, the carboxy terminal amino acid residue, or at an internal amino acid residue, for example, by a bond between a chemical moiety and a side chain of an internal amino acid residue of a peptide. Derivative peptide compounds useful in the methods of the invention also include any peptide conservative amino acid substitutions, addition of protective or capping groups on reactive moieties, and other changes that do not adversely destroy the activity of the peptide compound to treat or prevent pulmonary inflammation. [0026] An "amino terminal capping group" of a peptide compound described herein is any chemical compound or moiety that is linked, preferably covalently, to the amino terminal amino acid residue of a peptide compound. An amino terminal capping group may be useful to inhibit or prevent intramolecular cyclization or intermolecular polymerization, to promote transport of a peptide compound across the blood-brain barrier, to prevent degradation of the peptide compound, or to provide a combination of these properties. A peptide compound that is useful in this invention and that possesses an amino terminal capping group may possess other beneficial activities as compared with the uncapped peptide compound, such as enhanced efficacy, reduced side effects, enhanced hydrophilicity, or enhanced hydrophobicity. Examples of amino terminal capping groups of peptide compounds useful in the invention include, but are not limited to, 1 to 6 lysine residues, 1 to 6 arginine residues, a combination of arginine and lysine residues ranging from 2 to 6 residues, a methionine group (including methionine or methionine sulfoxide), urethanes, urea compounds, a lipoic acid ("Lip"), a palmitic acid moiety (i.e., palmitoyl group, "Palm"), glucose-3-O-glycolic acid moiety ("Gga"), a thyronine group (e.g., non-substituted thyronine, monoiodothyronine, diiodothyronine, triiodothyronine, tetraiodothyronine), and an acyl group that is covalently linked to the amino terminal amino acid residue of the peptide. Such acyl groups useful in the compositions of the invention may have a carbonyl group and a hydrocarbon chain that ranges from one carbon atom (e.g., as in an acetyl moiety) to up to 25 carbons (as in a docosahexaenoyl moiety, "DHA", which has a hydrocarbon chain that contains 22 carbons). Furthermore, the carbon chain of the acyl group may be saturated, as in a palmitoyl group, or unsaturated. It should be understood that when an acid or abbreviation for an acid (such as DI-LA, Palm, or Lip) is recited as an amino terminal capping group, the resultant peptide compound is the condensed product of the uncapped peptide and the acid. It should also be understood that the "amino terminal capping group" may comprise any combination of the chemical entities recited above. [0027] A "carboxy terminal capping group" of a peptide compound described herein is any chemical compound or moiety that is linked, preferably covalently, to the carboxy terminal amino acid residue of the peptide compound. A carboxy terminal capping group may be useful to inhibit or prevent intramolecular cyclization or intermolecular polymerization, to promote transport of the peptide compound across the blood-brain barrier, to prevent degradation of the peptide compound, or to provide a combination of these properties. A peptide compound that is useful in the methods of this invention and that possesses a carboxy terminal capping group may possess other beneficial activities as compared with the uncapped peptide, such as enhanced efficacy, reduced side effects, enhanced hydrophilicity, or enhanced hydrophobicity. [0028] Carboxy terminal capping groups that are particularly useful in the peptide compounds described herein include primary or secondary amines that are linked by an amide bond to the α-carboxyl group of the carboxy terminal amino acid of the peptide compound. Other carboxy terminal capping groups useful in the invention include aliphatic primary and secondary alcohols and aromatic phenolic derivatives, including flavenoids, with C₁ to C₂₆ carbon atoms, which form esters when linked to the carboxylic acid group of the carboxy terminal amino acid residue of a peptide compound described herein.

[0029] The terms "treat," "treating" and "treatment" refer to a method of alleviating or eliminating a disease, its attendant symptoms, and/or the cause of the disease itself. [0030] The terms "prevent," "preventing" and "prevention" refer to a method of delaying or precluding the onset of a disease and/or its attendant symptoms, barring a subject from acquiring a disease or reducing a subject's risk of acquiring a disease.

[0031] "Effective amount" refers to the amount of a compound that will elicit the biological or medical response of a cell, tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. "Pharmaceutically effective amount" or "therapeutically effective amount" may further include the amount of a peptide compound that, when administered, is sufficient to prevent development of, or alleviate to some extent, or eliminate, one or more of the symptoms of the condition or disorder being treated. [0032] "Pharmaceutical composition" refers to a composition that is suitable for medical administration to a subject for treating or preventing a disease or condition in the subject in need thereof. The pharmaceutical composition may contain one or more pharmaceutically acceptable carriers, excipients, adjuvants, diluents, vehicles, or the like.

[0033] A "subject" is defined herein to include animals such as mammals, including, but not limited to, primates, humans, cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like. In one embodiment, the subject is a human.

A. Methods Of Treating Or Preventing Diseases Affecting the Cardiac,

Pulmonary and Nervous Systems

[0034] The present invention provides methods of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof comprising administering to the subject an effective amount of at least one peptide compound capable of suppressing, inhibiting or preventing the disease.

[0035] The peptide compounds which are applicable in the present invention contain fewer than 20, and, in order of increasing preference, less than about 18, 15, 13, 9, 8, 7, 6, 5, 4 and 3 amino acids in length. Thus, one embodiment of the present invention is a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound, which the peptide compound contains fewer than 20 amino acids or, in order of increasing preference, less than about 18, 15, 13, 9, 8, 7, 6, 5, 4 or 3 amino acids in length, and more than 1 amino acid. [0036] In particular, the present invention describes a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound, wherein the peptide compound has the formula:

R₁ Xaai GIy Xaa₃ Xaa₄ Xaa₅ Xaa₆ Xaa₇ R₂, wherein R₁ is absent or is an amino terminal capping group of the peptide compound; Xaai and Xaa₃ are, independently, aspartic acid or asparagine; Xaa₄ is absent or GIy; Xaa₅ is absent, Asp, or Phe; Xaa₆ is absent, Ala, or Phe; Xaa₇ is absent or Ala; and R₂ is absent or is a carboxy terminal capping group of the peptide compound.

[0037] In particular, the present invention describes a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound, wherein the peptide compound is selected from a group consisting of:

R₁ Asp GIy Asp R₂;

R₁ ASp GIyASn R₂;

R₁ Asn GIy Asn R₂;

R₁ ASn GIyASp R₂;

R₁ Asn GIy Asp GIy R₂ (SEQ ID NO:1);

R₁ Asp GIy Asp GIy Asp R₂ (SEQ DD NO:2);

R₁ Asp GIy Asn GIy Asp R₂ (SEQ ID NO:3);

R₁ Asn GIy Asn GIy Asp R₂ (SEQ ID NO:4);

R₁ Asn GIy Asp GIy Asp R₂ (SEQ ID NO:5);

Ri Asp GIy Asp GIy Phe Ala R₂ (SEQ ID NO:6);

R₁ Asp GIy Asn GIy Phe Ala R₂ (SEQ ID NO:7);

R₁ Asn GIy Asn GIy Phe Ala R₂ (SEQ ID NO:8); R₁ Asn GIy Asp GIy Phe Ala R₂ (SEQ ID NO:9);

R₁ Asp GIy Asp GIy Asp Phe Ala R₂ (SEQ TD NO: 10);

R₁ Asp GIy Asn GIy Asp Phe Ala R₂ (SEQ ID NO: 11);

Ri Asn GIy Asn GIy Asp Phe Ala R₂ (SEQ ID NO:12); and

Ri Asn GIy Asp GIy Asp Phe Ala R₂ (SEQ ID NO: 13); wherein R₁ is absent or is an amino terminal capping group of the peptide compound; and R₂ is absent or is a carboxy terminal capping group of the peptide compound.

[0038] hi another embodiment, the invention provides a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound, wherein the peptide compound has the formula:

Ri Asp GIy Xaa₃ Xaa₄ Xaa₅ R₂, wherein Ri is absent or is an amino terminal capping group; Xaa₃ is GIu or Leu; Xaa₄ is Ala or GIu; Xaa₅ is absent, Leu, or Ala; and R₂ is absent or is a carboxy terminal capping group of the peptide compound.

[0039] The peptide compounds include, but are not limited to, those selected from the group consisting of:

R₁ Asp GIy GIu Ala R₂ (SEQ ID NO: 14);

R₁ Asp GIy GIu Ala Leu R₂ (SEQ ID NO: 15); and

R₁ Asp GIy Leu GIu Ala R₂ (SEQ ID NO: 16); wherein R₁ is absent or is an amino terminal capping group of the peptide compound; and R₂ is absent or is a carboxy terminal capping group of the peptide compound. In an embodiment, the Ri is an acetyl amino terminal capping group.

[0040] In yet another embodiment, the invention provides a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of a peptide compound having the formula:

R₁ Xaa! Xaa₂ Asp GIy Xaa₅ Xaa₆ Xaa₇ Xaa₈ Xaag Xaa₁₀ Xaaπ R₂; wherein R₁ is absent or is an amino terminal capping group; Xaa_ls is absent or any amino acid; Xaa₂ is absent or any amino acid; Xaa₅ is GIu or Leu; Xaa₆ is Ala or GIu; Xaa₇ is absent, Leu, or Ala; Xaa₈ is absent or is any amino acid; Xaa₉ is absent or is any amino acid; Xaa₁₀ is absent or is any amino acid; Xaaπ is absent or is any amino acid; and R₂ is absent or is a carboxy terminal capping group of the peptide compound.

[0041] In one embodiment, the invention provides a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound, wherein the peptide has any of the following amino acid "core sequences":

Asp GIy;

Asp GIy GIu;

Asp GIy GIu Ala (SEQ ID NO: 14);

Asp GIy GIu Ala Leu (SEQ ID NO: 15); and

Asp GIy Leu GIu Ala (SEQ ID NO: 16); wherein the peptide compound may contain one or more additional amino acids linked at the amino terminal and/or carboxy terminal amino acids of a "core sequence" of amino acids, provided such modification does not destroy the desired activity of the peptide compound to treat or prevent a disease affecting the cardiac, pulmonary and/or nervous system in a subject. In a further embodiment, the peptide compound contains one to six additional amino acids. [0042] In yet another embodiment, the present invention describes a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of a peptide compound having the formula:

R₁ Xaa_t Xaa₂ Xaa₃ R₂; wherein Rj is absent or is an amino terminal capping group; Xaai is Asp, Asn, GIu, GIn, Thr, or Tyr; Xaa₂ is absent or any amino acid (i.e., is variable); Xaa₃ is absent or is Asp, Asn, GIu, Thr, Ser, GIy, or Leu; and R₂ is absent or is a carboxy terminal capping group of the peptide compound. In one embodiment, Xaa₂ is selected from the group consisting of VaI, GIy, GIu, and GIn. In another embodiment, the method may comprise administering to the subject an effective amount of a peptide compound selected from the group consisting of:

R₁ Asp Ala R₂;

R₁ Asn Ala R₂;

R₁ Asn GIy R₂;

R₁ GIu GIy R₂;

R₁ GIu Ala R₂;

R₁ GIn GIyR₂;

R₁ GIn Ala R₂;

R₁ Thr VaI Ser R₂;

R₁ Asp GIy Asp R₂; and

R₁ ASn GIyASn R₂; wherein R₁ is absent or is an amino terminal capping group; and R₂ is absent or is a carboxy terminal capping group of the peptide compound.

[0043] Additional peptide compounds include but are not limited to,

Lip Asp GIy; Lip Asp Ala; Lip GIu GIy;

Lip GIu Ala;

Ac Asp GIy;

Ac Asp Ala;

Ac GIu GIy;

Ac GIu Ala;

T₃ Asp GIy;

T₃ Asp Ala;

T₃ GIu GIy;

T₃ GIu AIa;

Ac T₃ Asp GIy;

Ac T₃ Asp Ala;

Ac T₃ GIu GIy;

Ac T₃ GIu Ala;

Lip T₃ Asp GIy;

Lip T₃ Asp Ala;

Lip T₃ GIu GIy;

Lip T₃ GIu Ala; wherein Lip is lipoic acid group, in either oxidized or reduced form; Ac is an acetyl group; T₃ is triiodothyronine; Ac T₃ is acylated triiodothyronine and Lip T₃ is triiodothyronine linked to a lipoic moiety. These peptide compounds may also contain a carboxy terminal capping group, such as a primary or secondary amino group in an amide linkage to the carboxy terminal amino acid.

[0044] In yet another embodiment, the present invention describes a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of a peptide compound having the formula:

R₁ Xaa_! Xaa₂ Xaa₃ R_2; wherein Ri is absent or is an amino terminal capping group; Xaai is Met or methionine sulfoxide; Xaa₂ is GIu, GIn, Asn, or Asp; Xaa₃ is Ala or GIy; and R₂ is absent or is a carboxy terminal capping group.

[0045] Examples of peptide compounds include, but are not limited to, those having the following formulas:

R₁ Met GIu Ala R₂;

R₁ Met GIu GIy R₂;

R₁MCtGInGIyR₂;

R₁MCtGInAIaR₂;

Ri Met Asp Ala R₂;

R₁ Met AspGlyR₂;

Ri Met Asn Ala R₂;

R₁ Met Asn GIy R₂;

R₁Me^O)GIuAIaR₂;

RiMet(O)GluGlyR₂;

RiMet(O)GlnGlyR₂;

RiMCt(O)GInAIaR₂;

Ri Met(O) Asp Ala R₂;

RiMet(O)AspGlyR₂;

R₁ Met(O) Asn Ala R₂; and

RiMet(O)AsnGlyR₂; wherein R₁ is absent or is an amino terminal capping group; Met(O) is methionine sulfoxide; and R₂ is absent or is a carboxy terminal capping group. Methionine sulfoxide may be prepared by treating a methionine-containing peptide with DMSO.

[0046] In yet another embodiment, the invention provides a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, comprising administering to the subject an effective amount of a peptide compound having the formula:

R₁ Xaai Xaa₂ Met Thr Leu Thr GIn Pro R₂ (SEQ ID NO: 17), wherein R₁ is absent or is an amino terminal capping group; Xaai is absent or Ser; Xaa₂ is absent or Lys; and R₂ is absent or is a carboxy terminal capping group of the peptide compound. Additional peptide compounds are represented by the following formula:

Ri Met ThrXaa₃ R₂; wherein Ri is absent or is an amino terminal capping group; Xaa₃ is Asn, Asp, GIu, GIn, Thr, or Leu; and R₂ is absent or is a carboxy terminal capping group. [0047] Peptide compounds are also represented by the following formulas:

R₁ Met Thr Leu Thr GIn Pro R₂ (SEQ ID NO: 17); and

R₁ Ser Lys Met Thr Leu Thr GIn Pro R₂ (SEQ ID NO: 18); wherein Rj is absent or is an amino terminal capping group; and R₂ is absent or is a carboxy terminal capping group.

[0048] Another embodiment of the present invention is a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof, using a peptide compound having the formula:

R₁ Leu Xaa₂ Xaa₃ R₂; wherein Ri is absent or is an amino terminal capping group; Xaa₂ is any amino acid; Xaa₃ is GIn, GIy or Tyr; and R₂ is absent or is a carboxy terminal capping group. [0049] Peptide compounds of the present invention are also represented by the following formula:

R₁ Leu Thr GIn R₂;

R₁ Leu Thr GIy R₂; and

R₁ Leu Thr Tyr R₂; wherein Ri is absent or is an amino terminal capping group; and R₂ is absent or is a carboxy terminal capping group.

[0050] In addition to the peptide compounds described above, additional peptide compounds that may be applicable in the present invention for treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof include, but are not limited to, the following:

R₁ Asn Ser Thr R₂;

R₁ Phe Asp GIn R₂;

R₁ GIn Tyr Lys Leu GIy Ser Lys Thr GIy Pro GIy GIn R₂ (SEQ ID NO: 19);

R₁ GIn Thr Leu GIn Phe Arg R₂ (SEQ ID NO:20);

R₁ GIn Tyr Ser He GIy GIy Pro GIn R₂ (SEQ ID NO:21);

R₁ Ser Asp Arg Ser Ala Arg Ser Tyr R₂ (SEQ ID NO:22); and

R₁ Asp GIy Asp GIy Asp Phe Ala He Asp Ala Pro GIu R₂ (SEQ ID NO:23); wherein R₁ is absent or is an amino terminal capping group; and R₂ is absent or is a carboxy terminal capping group.

[0051] The peptide compounds useful in the methods of the present invention include derivative peptides containing one or more chemical moieties other than amino acids that are covalently attached at the amino terminal amino acid residue, the carboxy terminal amino acid residue, or at an internal amino acid residue by a bond between a chemical moiety and a side chain of an internal amino acid residue of the peptide. Thus, another embodiment of the present invention is methods of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof using an effective amount of a peptide compound described herein, wherein the peptide compound contains one or more chemical moieties. [0052] The peptide compounds useful in the methods of the invention also include any peptide having conservative amino acid substitutions, and/or protective or capping groups on reactive moieties, and other changes that do not adversely destroy the activity of the peptide compound to treat or prevent the diseases described herein.

[0053] As noted above, the peptide compounds described herein may contain an amino terminal capping group. Various amino terminal capping groups well-known in the field of peptide chemistry may be used in the present invention. Such amino terminal capping groups may play various functions, such as enhancing the efficacy of a peptide compound, protecting a peptide compound from degradation, and assisting administration of a peptide compound. Examples of such amino terminal capping groups may include, but are not limited to, a lipoic acid moiety (Lip, in reduced or oxidized form); a methionine group (methionine or methionine sulfoxide), a glucose-3-O-glycolic acid moiety (Gga); 1 to 6 lysine residues; 1 to 6 arginine residues; a combination of arginine and lysine residues ranging from 2 to 6 residues; a thyronine group; an acyl group of the formula R₃-CO-, where CO is a carbonyl group, and R₃ is a hydrocarbon chain having from 1 to 25 carbon atoms, and more preferably 1 to 22 carbon atoms, and where the hydrocarbon chain may be saturated or unsaturated, branched or unbranched, or any combination thereof.

[0054] In a preferred embodiment, the peptide compound may contain a lipoic acid moiety as an amino terminal capping group. Such a lipoic acid moiety may be in a reduced or oxidized form.

[0055] In another embodiment, the peptide compound may contain a thyronine group as the amino terminal capping group. As described above, a thyronine group may include, but is not limited to, a monoiodo-, diiodo-, triiodo-, or tetraiodothyronine or a thyronine group having no iodine substitutions. Such a thyronine group may also be an acylated thyronine group or a thyronine group linked to a lipoic group. In one embodiment, the triiodothyronine is a 3,5,3'- triiodothyronine, an acylated triiodothyronine group, or a lipoic triiodothyronine group. [0056] In yet another embodiment, the peptide compounds of the present invention may contain an acyl group as the amino terminal capping group, hi a further embodiment, the acyl group is an acetyl or a fatty acyl group. The acyl group may be selected from a group of acyl groups derived from a variety of well known acids, including but not limited to, acetic acid (acetyl), palmitic acid (palmitoyl), and docosahexaenoic acid (docosahexaenoly). The amino terminal capping group may include an acyl group and a thyronine group covalently linked together.

[0057] The peptide compounds of the present invention may also include a carboxy terminal capping group. Examples of carboxy terminal capping groups include, but are not limited to, primary or secondary amines.

[0058] Another embodiment of the present invention is a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof by administering two or more of the peptide compounds described herein. Multiple peptide compounds may be administered simultaneously or sequentially, and may also be incorporated into a single therapy.

[0059] The peptide compounds of the present invention may also be used in conjunction with other agents or compounds which are useful for treating, preventing or alleviating a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof. Such other agents or compounds may be administered by a route and in an amount commonly used thereof, simultaneously or sequentially, with one or more of the compounds of the invention. [0060] When multiple peptide compounds are used or a peptide compound of the present invention is used contemporaneously with one or more other agents or drugs, a pharmaceutical composition containing multiple peptide compounds or other agents or drugs in addition to the peptide compound of the invention may be prepared and administered. Thus, the present invention further provides a method of treating or preventing a disease affecting the cardiac, pulmonary and/or nervous system in a subject in need thereof by administering a pharmaceutical composition that includes a therapeutically effective amount of at least one peptide compound of the present invention and one or more other agents or compounds that are useful for treating, preventing or alleviating a disease or condition.

[0061] As described above, it is well known that pulmonary inflammation is associated with various diseases or conditions. Examples of such diseases and conditions may include, but are not limited to, pulmonary diseases, allergic reactions, physical trauma to pulmonary tissues, infection of the pulmonary system, and pulmonary conditions due to smoking and inhalation of noxious particles. Thus, another embodiment of the present invention is a method of treating or preventing a disease or condition in a subject in need thereof, wherein the disease or condition is selected from a group consisting of pulmonary disease, allergic reactions, trauma to pulmonary tissues, pulmonary infections and pulmonary conditions due to smoking or inhalation of noxious particles.

[0062] Furthermore, it is known that pulmonary inflammation is associated with various respiratory diseases and conditions. Examples of such respiratory diseases and conditions include, without limitation, asthma, bronchiectasis, broncholithiasis, cystic fibrosis, chronic bronchitis, chronic obstructive pulmonary disease, chronic granulomatous lung disease (i.e. Sarcoid), emphysema, eosinophilic pneumonias, hypersensitivity pneumonitis, idiopathic pulmonary fibrosis, and pneumonia. Thus, in another embodiment, the present invention describes a method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of at least one peptide compound described herein, wherein the disease or condition is selected from a group consisting of asthma, bronchiectasis, broncholithiasis, cystic fibrosis, chronic obstructive pulmonary disease (including chronic bronchitis, emphysema and chronic obstructive bronchitis), chronic granulomatous lung disease, eosinophilic pneumonias, hypersensitivity pneumonitis, idiopathic pulmonary fibrosis, and pneumonia.

[0063] In another embodiment, the methods of the invention are useful for treating or preventing pulmonary inflammation associated with allergic reactions affecting the pulmonary system. The allergic reactions may result from exposure to foreign allergens or autoimmune responses.

[0064] In yet another embodiment, the methods of the invention may be used to treat or prevent pulmonary inflammation associated with wounds or injuries to pulmonary tissues due to physical trauma. Such wounds or injuries may be the result of an accident or a surgical procedure.

[0065] The methods of the invention are also useful for treating or preventing pulmonary inflammation associated with respiratory infections such as those seen in patients with a cold or flu.

[0066] In another embodiment, the methods of the invention may be used to treat or prevent pulmonary inflammation associated with respiratory dysfunctions resulting from exposure to asbestos, dusts, toxic chemicals, and other inhalation of hazardous entities in the environment.

[0067] In another embodiment, the methods of the invention may be used to treat or prevent pulmonary inflammation associated with respiratory disorders resulting from smoking.

Smoking often causes pulmonary inflammation in people. The peptide compounds of the invention may be administered to people who smoke cigarettes in order to therapeutically treat or to prophylactically prevent pulmonary inflammation.

[0068] In a preferred embodiment, the methods of the invention are used to treat or prevent asthma. Pulmonary inflammation is typically associated with asthma and is a primary contributor to the symptoms of asthma. Asthma is an episodic disease, wherein acute onset of the disease is interspersed with symptom-free periods. Typically, most attacks are short-lived, lasting minutes to hours. After an attack, the patient seems to recover completely. However, under certain circumstances, acute episodes of asthma may be fatal. Many factors may trigger the acute onset of asthma, including, for example, respiratory infections, pollution, and emotional stress. In one embodiment, the methods of the invention are used to prevent the onset of asthma before, during or after an individual's exposure to stimuli that may trigger an acute asthma attack.

[0069] In another preferred embodiment, the methods of the invention are used to treat or prevent chronic obstructive pulmonary disease. Patients with chronic obstructive pulmonary disease exhibit inflammation in their respiratory tracts. Pulmonary inflammation in these patients may aggravate the symptoms of the disease and cause severe complications. The methods of the invention are useful for treating or inhibiting the disease.

[0070] As described above, the compositions of the present invention are also suitable for treating or preventing a variety of diseases and conditions affecting the cardiac and nervous systems. Accordingly, another embodiment of the present invention is a method of treating or preventing a disease or condition in a subject, preferably a human, in need thereof, comprising administering to the subject a pharmaceutical composition, including an effective amount of at least one peptide compound, a pharmaceutically acceptable carrier and, optionally, DMSO. Examples of such diseases or conditions include, but are not limited to, cognitive degeneration, senility, Tardive dyskinesia, head trauma, brain and/or spinal cord trauma, oxygen toxicity, Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis, Alzheimer's disease, diabetes, ulcerative colitis, leukemia and other cancers characterized by elevation of ROS or free radicals, Downs syndrome, macular degeneration, cataracts, schizophrenia, epilepsy, septic shock, polytraumatous shock, burn injuries, and radiation-induced elevation of ROS. [0071] Another preferred embodiment of the present invention is a method of treating or preventing ischemia in a subject, including administering to the subject a pharmaceutical composition, wherein the pharmaceutical composition comprises an effective amount of at least one peptide compound. Ischemic cerebrovascular disease, anoxic-ischemic encephalopathy, ischemic heart disease, ischemic renal disease, intestinal ischemia, cerebral ischemia and myocardial ischemia are examples of diseases which may be treated or prevented according to the present invention.

B. Synthesis and Preparation of Peptide Compounds Useful in the Invention

[0072] The synthesis and preparation of the peptide compounds useful in the methods of the invention have been previously described in international patent applications published as WO 01/36454, WO 02/092781, WO 02/096360, and WO 03/066814, which are incorporated herein by reference in their entireties. Methods for synthesizing and purifying the peptide compounds of the invention are briefly described below.

[0073] The peptide compounds of the invention can be made using standard methods or obtained from a commercial source. Synthesis of the peptides may be accomplished using conventional techniques, including solid-phase peptide synthesis, and solution-phase synthesis. Peptides may also be synthesized using various recombinant nucleic acid technologies, however, given their relatively small size and the state of peptide synthesis technology, a direct synthesis is usually utilized and solid-phase synthesis is most often employed.

[0074] In solid-phase synthesis, for example, a suitably protected amino acid residue is attached through its carboxyl group to a derivatized, insoluble polymeric support, such as cross linked polystyrene or polyamide resin. "Suitably protected" refers to the presence of protecting groups on both the ce-amino group of the amino acid, and on any side chain functional groups. Side chain protecting groups are generally stable to the solvents, reagents, and reaction conditions used throughout the synthesis and are removable under conditions which do not affect the final peptide product. Stepwise synthesis of the polypeptide is carried out by the removal of the N-ρrotecting group from the initial (i.e., carboxy terminal) amino acid, and coupling thereto of the carboxyl end of the next amino acid in the sequence of the polypeptide. This amino acid is also suitably protected. The carboxyl group of the incoming amino acid can be activated to react with the N-terminus of the bound amino acid by formation into a reactive group such as formation into a carbodiimide, a symmetric acid anhydride, or an "active ester" group, such as hydroxybenzotriazole or pentafluorophenyl esters.

[0075] The preferred solid-phase peptide synthesis methods include the BOC method, which utilizes tertbutyloxycarbonyl as the α-amino protecting group, and the FMOC method, which utilizes 9 fluorenylmethloxycarbonyl to protect the oamino of the amino acid residues. Both methods are well known by those of skill in the art (see, Stewart et al., In Solid-Phase Peptide Synthesis (W. H. Freeman Co., San Francisco 1989); Merrifield, J. Am. Chem. Soc, 85:2149-2154 (1963); Bodanszky and Bodanszky, In The Practice of Peptide Synthesis (Springer- Verlag, New York 1984)).

[0076] Amino terminal and carboxy terminal capping groups, if desired, may be added during or after peptide synthesis, depending on the specific moiety used as a capping group. For example, if the capping group is one or more amino acids, then such residues are simply incorporated into the protocol for synthesizing the peptide. If the capping group is not an amino acid, such as an acyl or amide group, it may be added after peptide synthesis using standard condensation or conjugation methods. [0077] Peptide compounds described herein may also be prepared commercially by companies providing peptide synthesis as a service (e.g., BACHEM Bioscience, Inc., King of Prussia, PA; AnaSpec, Inc., San Jose, CA). Automated peptide synthesis machines, such as manufactured by Perkin-Elmer Applied Biosystems, also are available.

[0078] Peptide compounds useful in the methods of the invention may also be prepared and used in a salt form. Typically, a salt form of a peptide compound will exist by adjusting the pH of a composition comprising the peptide compound with an acid or base in the presence of one or more ions that serve as counter ion to the net ionic charge of the peptide compound at the particular pH. Various salt forms of the peptide compounds described herein may also be formed or interchanged by any of the various methods known in the art, including the use of various ion exchange chromatography methods. Cationic counter ions that may be used in the compositions described herein include, but are not limited to, amines, such as ammonium ion; metal ions, especially monovalent or divalent ions of alkali metals (e.g., sodium, potassium, lithium, cesium), alkaline earth metals (e.g., calcium, magnesium, barium), transition metals (e.g., iron, manganese, zinc, cadmium, molybdenum), other metals (e.g., aluminum); and combinations thereof. Anionic counter ions that may be used in the compositions described herein include, but are not limited to, chloride, fluoride, acetate, trifluoroacetate, phosphate, sulfate, carbonate, citrate, ascorbate, sorbate, glutarate, ketoglutarate, and combinations thereof. Trifluoroacetate salts of peptide compounds described herein are typically formed during purification in trifluoroacetic acid buffers using high-performance liquid chromatography (HPLC). Trifluoroacetate salt forms of peptide compounds are generally not suited for in vivo use, although they may be conveniently used in various in vitro cell culture studies or assays performed to test the activity or efficacy of a peptide compound of interest. The peptide compound may then be converted from the trifluoroacetate salt (e.g., by ion exchange methods) or synthesized as a salt form that is acceptable for pharmaceutical use. [0079] The peptide compounds described herein may contain a peptide to which additional modifications have been made, such as the addition of chemical moieties at the amino terminal and/or carboxy terminal amino acid residues of the peptide, conservative amino acid substitutions, or modifications of side chains of internal amino acid residues of the peptide that do not destroy the desired activity of the peptide. It has been observed that intramolecular cyclization, and some intermolecular polymerizations of peptides tend to inactivate or decrease the activity of the peptides. Accordingly, the most useful peptide compounds are the least susceptible to cyclization reactions or undesired polymerization reaction or conjugation with other peptide compound molecules.

[0080] Preferred amino terminal capping groups include a lipoic acid moiety which can be attached by an amide linkage to the α-amino group of the amino terminal amino acid of a peptide. An amino terminally linked lipoic acid moiety may be in its reduced form where it contains two sulfhydryl groups or in its oxidized form in which the sulfhydryl groups are oxidized and form an intramolecular disulfide bond and, thereby, a heterocyclic ring structure. Another amino terminal capping group useful in preparing peptide compounds of the invention is a glucose-3-O-glycolic acid moiety, which can be attached in an amide linkage to the oamino group of the amino terminal amino acid of a peptide compound. The glucose moiety may also contain further modifications, such as an alkoxy group replacing one or more of the hydroxyl groups on the glucose moiety.

[0081] Another example of an amino terminal capping group useful in the peptide compounds described herein is an acyl group, which can be attached in an amide linkage to the α-amino group of the amino terminal amino acid residue of a peptide compound. The acyl group has a carbonyl group linked to a saturated or unsaturated (mono- or polyunsaturated), branched or unbranched, hydrocarbon chain. The acyl group preferably is acetyl or a fatty acid. The fatty acid used as the acyl amino terminal capping group may contain a hydrocarbon chain that is saturated or unsaturated and that is either branched or unbranched. Preferably, the hydrocarbon chain of the acyl group is 1 to 25 carbon atoms in length, and more preferably the length of the hydrocarbon chain is 1-22 carbon atoms in length, such as in DHA. For example, fatty acids that are useful in their corresponding acyl form, as amino terminal capping groups linked to the peptide compounds of this invention include, but are not limited to: caprylic acid (C8:0), capric acid (C10:0), lauric acid (C 12:0), myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:l), C16:2, stearic acid (C18:0), oleic acid (C18:l), vaccenic acid (C18.-1-7), linoleic acid (C18:2-6), α-linolenic acid (C18:3-3), eleostearic acid (C18:3-5), /3-linolenic acid (C18:3-6), C18:4-3, gondoic acid (C20:l), C20:2-6, dihomo-γ-linolenic acid (C20:3-6), C20:4-3, arachidonic acid (C20:4-6), eicosapentaenoic acid (C20:5-3), docosenoic acid (C22:l), docosatetraenoic acid (C22:4-6), docosapentaenoic acid (C22:5-6), docosapentaenoic acid (C22:5-3), docosahexaenoic acid (C22:6-3), and nervonic acid (C24:l-9). Two fatty acids that may be used as acyl amino terminal capping groups for the peptide compounds described herein are a palmitoyl moiety and a docosahexaenoyl moiety.

[0082] In addition, in certain cases the amino terminal capping group may be a lysine residue or a polylysine peptide. In one embodiment, the polylysine peptide consists of two, three, four, five or six lysine residues. Longer polylysine peptides may also be used. Another amino terminal capping group that may be used in the peptide compounds described herein is an arginine residue or a polyarginine peptide. In one embodiment, the polyarginine peptide consists of two, three, four, five, or six arginine residues, although longer polyarginine peptides may also be used. An amino terminal capping group of the peptide compounds described herein may also be a peptide containing both lysine and arginine. Li one embodiment, the lysine and arginine containing peptide is two, three, four, five or six residue combinations of the two amino acids in any order, although longer peptides that contain lysine and arginine may also be used. Lysine and arginine containing peptides used as amino terminal capping groups in the peptide compounds described herein may be conveniently incorporated into whatever process is used to synthesize the peptide compounds to yield the derivatized peptide compound containing the amino terminal capping group.

[0083] Yet another example of an amino terminal capping group may be one or more methionine or methionine sulfoxide. Methionine sulfoxide may be prepared using methionine and DMSO. Methionine sulfoxide containing peptide compounds may be prepared by treating such methionine-containing peptide compounds with DMSO.

[0084] Yet another example of an amino terminal capping group useful in the invention is a thyronine group (i.e., thyronyl). The α-carboxyl group of a thyronine residue may be conjugated to an α-amino group of an amino acid residue by condensation to form a peptide bond, e.g., during standard Merrifield synthesis of a peptide compound. Thyronines useful as amino terminal capping groups of the peptide compounds described herein include but are not limited to thyronine having no iodine substitution ("T₀"), monoiodothyronine ("T₁,"), diiodothyronine ("T₂"), triiodothyronine ("T₃"), and tetraiodothyronine ("T₄"). Li the case of iodothyronines, a position that is substituted with iodine (iodinated) on one or both of the phenyl groups of a particular species of iodothyronine may vary, but preferably occurs at phenyl position 3, 3', 5, 5', or a combination thereof. Diagrams of several examples of thyronine groups useful in the invention are shown in Figure 1 in a form in which each thyronine group may be linked at its carbonyl atom to another molecule (e.g., linked in a peptide bond to the amino terminal amino group of a peptide). The 3, 3', 5, 5 '-tetraiodothyronine group (3, 3', 5, 5' T₄) is derived from the thyroxine hormone (also referred to as "thyroxine T₄"). The α-amino group of a thyronine may be free or linked to another moiety (e.g., blocked), as desired, using standard reactions known in the art, e.g., treatment with acetic anhydride or lipoic acid may be used to couple an acetyl group or a lipoic group to the α-amino group of thyronine. [0085] The peptide compounds useful in the methods of the invention may contain a carboxy terminal capping group. The primary purpose of this group is usually to prevent intramolecular cyclization, or inactivating intermolecular crosslinking, or polymerization. Cyclization, crosslinking, or polymerization of a peptide compound described herein may abolish all or so much of the activity of the peptide compound that it cannot be used in the therapeutic or prophylactic compositions and methods of the invention. However, as noted above, a carboxy terminal capping group may provide additional benefits to a peptide compound described herein, such as enhanced efficacy, reduced side effects, and/or other desirable biochemical properties. An example of a useful carboxy terminal capping group is a primary or secondary amine in an amide linkage to the carboxy terminal amino acid residue. Such amines may be added to the α-carboxyl group of the carboxy terminal amino acid of the peptide using standard amidation chemistry.

[0086] In addition, peptide compounds described herein may contain one or more D- amino acid residues in place of one or more L-amino acid residues, provided that the incorporation of the one or more D-amino acids does not abolish all, or so much of the activity of the peptide compound that it cannot be used in the methods of the invention. Incorporating D- amino acids in place of L-amino acids may advantageously provide additional stability to a peptide compound.

[0087] After being produced or synthesized, a peptide compound that is useful in the methods of the invention may be purified using methods known in the art. Such purification should provide a peptide compound of the invention in a state dissociated from significant or detectable amounts of undesired side reaction products, dissociated from unattached or unreacted moieties used to modify the peptide compound, and dissociated from other undesirable molecules, including but not limited to other peptides, proteins, nucleic acids, lipids, and carbohydrates. Standard methods of peptide purification may be employed to obtain isolated peptide compounds of the invention, including but not limited to various high pressure (or performance) liquid chromatography (HPLC) and non-HPLC peptide isolation protocols, such as size exclusion chromatography, ion exchange chromatography, phase separation methods, electrophoretic separations, precipitation methods, salting in/out methods, and immunochromatography.

[0088] A particularly preferred method of isolating peptide compounds useful in compositions and methods of the invention employs reversed phase HPLC, using an alkylated silica column such as C₄, C₈ or C₁₈-silica. A gradient mobile phase of increasing organic content is generally used to achieve purification, for example, acetonitrile in an aqueous buffer, usually containing a small amount of trifluoroacetic acid. Ion exchange chromatography can also be used to separate peptide compounds based on their charge.

[0089] The degree of purity of the peptide compound may be determined by various methods, including identification of a major large peak on HPLC. A peptide compound that produces a single peak that is at least 95% of the input material on an HPLC column is preferred. Even more preferable is a polypeptide that produces a single peak that is at least 97%, at least 98%, at least 99% or at least 99.5% of the input material on an HPLC column. [0090] In order to ensure that a peptide compound obtained using any of the techniques described above is the desired peptide compound for use in methods of the present invention, the compound's composition may be analyzed by any of a variety of analytical methods known in the art. Such composition analysis may be conducted using high resolution mass spectrometry to determine the molecular weight of the peptide. Alternatively, the amino acid content of a peptide can be confirmed by hydrolyzing the peptide in aqueous acid, and separating, identifying and quantifying the components of the mixture using HPLC, or an amino acid analyzer. Protein sequenators, which sequentially degrade the peptide and identify the amino acids in order, may also be used to determine the sequence of the peptide. Since some of the peptide compounds contain amino and/or carboxy terminal capping groups, it may be necessary to remove the capping group or the capped amino acid residue prior to a sequence analysis. Thin-layer chromatography (TLC) methods may also be used to authenticate one or more constituent groups or residues of a desired peptide compound. Purity of a peptide compound may also be assessed by electrophoresing the peptide compound in a polyacrylamide gel, followed by staining to detect the separated protein components in the gel. C. Preparation Of Pharmaceutical Compositions

[0091] The peptide compounds useful in the methods of the invention will treat or prevent any disease described herein in a subject when administered to a subject at various concentrations ranging from nanograms (ng) to milligrams (mg) of peptide compound per milliliter (ml). In some cases, the potency is similar to that exhibited by various hormones, such as luteinizing hormone releasing hormone (LHRH) or human growth hormone. Accordingly, the peptide compounds described herein may be prepared, stored, and used employing much of the available technology already applied to the preparation, storage, and administration of known therapeutic hormone peptides.

[0092] The methods of the invention employ a peptide compound, described herein, in a pharmaceutical composition for administration to a subject to treat or prevent any of the diseases described herein in such subject. Accordingly, the methods of this invention comprise the use of any of the peptide compounds described herein, or pharmaceutically acceptable salts thereof, as the active ingredient (also called the "pharmaceutical agent") of a pharmaceutical composition. Pharmaceutical compositions employed in the invention may further comprise one or more other pharmaceutically acceptable ingredients, including an excipient (a compound that provides a desirable property or activity to the composition, but other than or in addition to that of the active ingredient), a carrier, an adjuvant, a diluent, a vehicle, or the like. [0093] Pharmaceutical compositions of this invention can be administered to a subject, such as a mammal, and especially a human patient, in a manner similar to other therapeutic, prophylactic, and diagnostic agents, and especially compositions comprising therapeutic hormone peptides. The dosage to be administered, and the mode of administration will depend on a variety of factors including age, weight, sex, condition of the subject, and genetic factors, and will ultimately be decided by the attending physician or veterinarian. In general, the dosage required for diagnostic sensitivity or therapeutic efficacy will range from about 0.001 to 25.0 mg/kg of host body mass (also referred to herein as body weight).

[0094] Pharmaceutically acceptable salts of the peptide compounds useful in this invention include, for example, those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, malic, pamoic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, tannic, carboxymethyl cellulose, polylactic, polyglycolic, and benzenesulfonic acids. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts. Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N-(C_1-4 alkyl)₄ ⁺ salts.

[0095] This invention also envisions the "quaternization" of any basic nitrogen containing groups of a peptide compound disclosed herein, provided such quaternization does not destroy the ability of the peptide compound to treat or prevent any of the diseases described herein. Such quaternization may be especially desirable where the goal is to use a peptide compound containing only positively charged residues. For example, when charged amino acid residues are present in a peptide compound, they are preferably either all basic (positively charged) or all acidic (negatively charged) which prevents formation of cyclic peptide compounds during storage or use. Cyclic forms of the peptide compounds may be inactive and potentially toxic. Thus, a quaternized peptide compound is a preferred form of a peptide compound containing basic amino acids. Even more preferred is the quaternized peptide compound in which the carboxy terminal carboxyl group is converted to an amide to prevent the carboxyl group from reacting with any free amino groups to form a cyclic compound. Any basic nitrogen can be quaternized with any agent known to those skilled in the art, including, for example, lower alkyl halides, such as methyl, ethyl, propyl and butyl chloride, bromides and iodides; dialkyl sulfates, including dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; and aralkyl halides including benzyl and phenethyl bromides. Water or oil soluble or dispersible products may be obtained by such quaternization or acids such as acetic acid and hydrochloric acid. [0096] It should be understood that the peptide compounds described herein may be modified by appropriate functionalities to enhance selective biological properties, and in particular the ability to treat or prevent a disease affecting the cardiac, pulmonary and/or nervous system. Such modifications are known in the art and include those which increase the ability of the peptide compound to penetrate or be transported into a given biological system (e.g., circulatory system, lymphatic system), increase oral availability, increase solubility to allow administration by injection, alter the metabolism of the peptide compound, and alter the rate of excretion of the peptide compound. In addition, peptide compounds may be altered to a pro-drug form such that the desired peptide compound is created in the body of an individual as the result of the action of metabolic or other biochemical processes on the pro-drug. Such pro-drug forms typically demonstrate little or no activity in in vitro assays. Some examples of pro-drug forms may include ketal, acetal, oxime, and hydrazone forms of compounds, which contain ketone or aldehyde groups. Other examples of pro-drug forms include the hemi-ketal, hemi-acetal, acyloxy ketal, acyloxy acetal, ketal, and acetal forms.

[0097] Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene polyoxypropylene block polymers, polyethylene glycol and wool fat.

[0098] The pharmaceutical compositions used in the methods of this invention may be administered by a variety of routes or modes. These include, but are not limited to, parenteral, oral, intratracheal, sublingual, pulmonary, topical, rectal, nasal, buccal, vaginal, or via an implanted reservoir. Implanted reservoirs may function by mechanical, osmotic, or other means. The term "parenteral", as understood and used herein, includes intravenous, intracranial, intraperitoneal, paravertebral, periarticular, periostal, subcutaneous, intracutaneous, intra-arterial, intramuscular, intra articular, intrasynovial, intrasternal, intrathecal, and intralesional injection or infusion techniques. Such compositions are preferably formulated for parenteral administration, and most preferably for intravenous, intracranial, or intra-arterial administration. Generally, and particularly when administration is intravenous or intra-arterial, pharmaceutical compositions may be given as a bolus, as two or more doses separated in time, or as a constant or non-linear flow infusion.

[0099] The pharmaceutical compositions used in the invention may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. Such a suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3- butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long chain alcohol diluent or dispersant such as those described in Pharmacoplia Halselica. [00100] The pharmaceutical compositions of this invention may be administered in any orally acceptable dosage form including, but not limited to, aqueous solutions and suspensions, capsules, tablets, caplets, pills, oleaginous suspensions and solutions, syrups, and elixirs, hi the case of tablets for oral use, carriers, which are commonly used include lactose and cornstarch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. Capsules, tablets, pills, and caplets may be formulated for delayed or sustained release.

[00101] When aqueous suspensions are to be administered orally, the peptide compound is advantageously combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added. Formulations for oral administration may contain 10%-95% (weight/volume, w/v) active ingredient, and preferably 25%-70% (w/v). Preferably, a pharmaceutical composition for oral administration provides a peptide compound of the invention in a mixture that prevents or inhibits hydrolysis of the peptide compound by the digestive system, but allows absorption into the blood stream. [00102] The pharmaceutical compositions of this invention may also be administered in the form of suppositories for vaginal or rectal administration. These compositions can be prepared by mixing a peptide compound described herein with a suitable non-irritating excipient, which is solid at room temperature but liquid at body temperature, so that the composition will melt in a relevant body space to release the active ingredient. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols. Formulations for administration by suppository may contain 0.5%~l 0% (w/v) active ingredient, preferably l%-2% (w/v). [00103] Topical administration of the pharmaceutical compositions used in the methods of the invention may also be useful. For application topically, the pharmaceutical composition may be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the peptide compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the peptide compounds suspended or dissolved in a pharmaceutically suitable carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical composition may be formulated for topical or other application as a jelly, gel, or emollient, where appropriate. The pharmaceutical compositions used in this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topical administration may also be accomplished via transdermal patches. [00104] The pharmaceutical compositions employed in this invention may be administered by inhalation through the nose or mouth, in which case absorption may occur via the mucus membranes of the nose or mouth, or inhalation into the lungs. Such modes of administration typically require that the composition be provided in the form of a powder, solution, or liquid suspension, which is then mixed with a gas (e.g., air, oxygen, nitrogen, etc., or combinations thereof) so as to generate an aerosol or suspension of droplets or particles. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art. Pharmaceutical compositions of the invention may also be administered to a subject by inhalation.

[00105] The pharmaceutical compositions of the invention may also be formulated as slow release formulations. Such formulations may be administered by implantation, for example, subcutaneously or intramuscularly or by intramuscular injection. Such formulations may be prepared according to techniques well known in the art of pharmaceutical formulation, for example, as an emulsion in an acceptable oil, or ion exchange resins, or as sparingly soluble derivatives (e.g. sparingly soluble salts).

[00106] Pharmaceutical compositions of the invention may be packaged in a variety of ways appropriate to the dosage form and mode of administration. These include but are not limited to vials, bottles, cans, packets, ampoules, cartons, flexible containers, inhalers, and nebulizers. Such compositions may be packaged for single or multiple administrations from the same container. Kits, of one or more doses, may be provided containing the composition in dry powder or lyophilized form and an appropriate diluent, which are to be combined shortly before administration; and instructions for preparation and/or administration of the reconstituted or otherwise prepared pharmaceutical composition. The pharmaceutical composition may also be packaged in single use pre-filled syringes, or in cartridges for auto-injectors and needleless jet injectors. [00107] Multi-use packaging may require the addition of antimicrobial agents such as phenol, benzyl alcohol, meta-cresol, methyl paraben, propyl paraben, benzalcom^'um chloride, and benzethonium chloride, at concentrations that will prevent the growth of bacteria, fungi, and the like, but are non-toxic when administered to a patient.

[00108] Details concerning dosages, dosage forms, modes of administration and compositions are further discussed in a standard pharmaceutical text, such as Remington's

Pharmaceutical Sciences (1990), which is incorporated herein by reference.

[00109] The present invention is described in the following Examples, which are set forth to aid in the understanding of the invention, and should not be construed to limit in any way the scope of the invention as defined in the claims which follow thereafter.

EXAMPLES

Example 1

Synthesis Of Representative Peptide Compounds

[00110] Representative peptide compounds useful in the methods of the invention were synthesized and characterized as indicated below.

[00111] CMX-I: a peptide compound that is a polypeptide having the amino acid sequence Asp GIy Asp GIy Asp Phe Ala He Asp Ala Pro GIu (SEQ ID NO:23).

[00112] CMX-2: a peptide compound having the formula [Ac] Asp GIy GIu Ala (SEQ ID

NO: 14).

[00113] CMX-3: a peptide compound that is a dipeptide having the amino acid sequence

Asp GIy.

[00114] CMX-4: a peptide compound having the formula R₁ Asp GIy, wherein Ri is the iodine-substituted thyronine amino terminal capping group, 3, 5, 3 '-triiodothyronine (3, 5, 3' T₃ shown in Figure 1). [00115] CMX-5: a peptide compound having the formula R₁ Asp GIy, wherein R₁ is a lipoic group.

[00116] CMX-6: a peptide compound having the formula R₁ GIu Ala, wherein R₁ is a lipoic group.

[00117] CMX-8: a peptide compound having the amino acid sequence GIu Ala.

[00118] CMX-9: a peptide compound having the formula R₁ Asp GIy, wherein R₁ is the acylated iodine-substituted thyronine amino terminal capping group, 3, 5, 3 '-triiodothyronine which is acylated.

[00119] CMX-Il: a peptide compound having the formula R₁ GIu Ala, wherein R₁ is lipoic acid moiety.

[00120] The following representative compounds may be synthesized and characterized as indicated below.

[00121] CMX-7: a peptide compound having the formula R₁ GIu Ala, wherein R₁ is the iodine-substituted thyronine amino terminal capping group, 3, 5, 3 '-triiodothyronine (3, 5, 3' T₃ shown in Figure I).

[00122] CMX-10: a peptide compound having the formula R₁ GIu Ala, wherein R₁ is the acylated iodine-substituted thyronine amino terminal capping group, 3, 5, 3 '-triiodothyronine which is acylated.

[00123] Amino terminal capping groups, as indicated by the bracketed groups "[DHA]-",

"[Lip]-", and "[Ac]", which represent an all cis-docasahexaenoic moiety, a lipoic acid moiety, and an acetyl moiety, respectively, could be attached to the α-amino group of the amino terminal amino acid residue of the indicated peptide compounds (Shashoua and Hesse, Life Sci. 58:1347-

1357 (1996)). The T₃ iodothyronine group was attached by condensation to form a peptide bond with the α-amino group of the amino terminal aspartic acid residue in the CMX-4 peptide compound during Merrifield synthesis. [00124] The peptide compounds were synthesized using standard procedures. The peptides were synthesized using the solid-phase Merrifield process (Merrifield, J. Am. Chem. Soc, 85:2149-2154 (1963)) or by a traditional solution phase process. The Merrifield process allows the synthesis of a peptide of a specific amino acid sequence bound on a polymeric resin. Each newly synthesized peptide was then released from the resin by treating with trifluoroacetic acid (TFA). The resultant trifluoroacetic acid peptide salt was purified by ether precipitation according to standard procedures (see, Groos and Meienhofer, In The peptides, analysis, synthesis, biology, vol. 2, (Academic Press, New York 1983)).

[00125] For amino terminal substituted peptides (i.e., peptides containing an acyl amino terminal capping group), each peptide was synthesized with blocked side chains using solid- phase Merrifield synthesis (see above). The bound peptide was then treated with an equimolar amount of an anhydride of one of the following acids: acetic acid, DHA, or lipoic acid, in the presence of 4-dimethylaminopyridine under argon atmosphere. The reaction was carried out for about three hours to obtain amino terminal coupling. Evidence of complete amino terminal coupling was obtained prior to peptide isolation. This was established by monitoring the ninhydrin staining properties of the resin bound peptides using standard procedures (Kaiser et al., Anal. Biochem., 34: 595-598 (1970)). The amino terminal coupled (capped) peptide molecule was then released from the resin by treatment with TFA and purified by precipitation with cold ether followed by HPLC using methanolic HCl (50:50) as the eluant.

[00126] The final peptide products were white solids after lyophilization. Structures were confirmed by amino acid analysis, by migration as a single peak on HPLC, and molecular weight determinations by mass spectrometry. For most applications, it was essential to remove residual TFA from the peptide compound. This was achieved by repeated dissolution of the peptide in glacial acetic acid followed by concentration in vacuum in rotary evaporator. Example 2 Experimental Protocols for Measuring Inflammation Induced by Urban Dusts in Rats

[00127] Urban dusts in vials (PMlO) was purchased from the National Institute of

Standards & Technology (NIST) in Gaithersburg, MD, U.S. A (reference no. SRM 1649a). The composition was resuspended in sterile saline to a working concentration of 1 mg/ml and sonicated in a waterbath for 5 minutes.

[00128] Sprague-Dawley male rats weighing between 300-400 grams were anaesthetized with Ketamine (Vetalar) and medetomidine (Dormitor) at doses 20% lower than recommended by the manufacturers. Rats were then intratracheally instilled with different concentrations of urban dusts (SRM 1649a solution) in a final volume of 250 ml. Following instillation, animals were given a reversal agent (anti-sedan agent) to counteract the anaesthetic effects. Anti-sedan agent was given at least 30 minutes after the anaesthetic injection.

[00129] Twenty-four hours after exposure, rats were sacrificed using an overdose of pentobarbital solution. Lungs were removed and bronchoalveolar lavages (BAL) were performed using 8 ml of sterile saline, which was repeated 4 times. The lavaged lungs were snap-frozen in liquid nitrogen for future analysis. The first lavage was collected separately to use for biochemical analysis, whereas lavages two through four were collected in the same Falcon tube. Following centrifuge at 1,200 rpm for 6 minutes, BAL from the first lavage was aliquoted into eppendorf tubes and stored at -80 degree for further analysis.

Example 3 Suppression of Inflammation Induced by Urban Dusts in Rats

[00130] The study was designed to instill male Sprague-Dawley rats with urban dusts (PM

10) and test whether the peptide compounds described herein were able to attenuate the inflammatory response by the rats to urban dusts. [00131] Initially, rats were exposed to PMlO at a concentration of 125 μg, 250 μg or 500 μg per rat for 24 hours to determine an optimal dose of PMlO suitable for inducing maximal inflammatory response in rats. As a control, rats were exposed to saline solution (vehicle). As shown in Figure 2, compared to rats exposed to saline solution alone, when exposed to different concentrations of PM 10, rats displayed an increase of neutrophils in the BAL fluid. The maximal inflammatory response was observed in rats instilled with PMlO at a concentration of 250 μg. The maximal level of inflammatory response when exposed to PM 10 at a concentration of 250 μg per rat was about 25%, as compared to 5% inflammation in rats when exposed to saline solution alone.

[00132] Rats were injected intravenously with CMX-Il (Lip GIu Ala) at a concentration of

1 mg/kg and saline solution, respectively, for three days consecutively. On day 3, rats were also instilled with 250 μg of PMlO for 24 hours. Rats were then sacrificed in accordance with the method of Example 2. As shown in Figure 3, exposure to CMX-11 led to up to 74% reduction of neutrophils in the BAL fluid.

Example 4

Treatment of Humans Suffering Pulmonary Inflammation

[00133] A randomized, double-blind, placebo controlled study is conducted.

Approximately 100 patients, both men and women between the ages of 20 and 50, with a diagnosis of pulmonary inflammation, are recruited for participation in the study. [00134] Patients are randomized for administration of a candidate peptide compound in the amount of 0.1 mg/day to 1000 mg/day, or a placebo for twelve weeks. Prior to randomization, patients are evaluated for pulmonary inflammation. The primary endpoint is a comparison between the treatment and placebo groups.

Example 5 Treatment of Human Patients Suffering Chronic Obstructive Pulmonary Disease [00135] A randomized, double-blind, placebo controlled study is conducted.

Approximately 100 patients, both men and women between the ages of 50 and 70, with a diagnosis of chronic obstructive pulmonary disease, are recruited for participation in the study. [00136] Patients are randomized for administration of a candidate peptide compound in the amount of 0.1 mg/day to 1000 mg/day, or a placebo for twelve weeks. Prior to randomization, patients are evaluated for chronic obstructive pulmonary disease. The primary endpoint is a comparison between the treatment and placebo groups.

[00137] While the foregoing invention has been described in some detail for purposes of clarity and understanding, it will be appreciated by one skilled in the art, from a reading of the disclosure, that various changes in form and detail can be made without departing from the true scope of the invention in the appended claims.

Claims

What is claimed is:

1. A method of treating or preventing pulmonary inflammation in a subject in need thereof, comprising administering to the subject a composition comprising an effective amount of at least one peptide compound, wherein the peptide compound is capable of suppressing pulmonary inflammation and contains at least one but less than twenty amino acids.

2. The method of claim 1, wherein the peptide compound contains at least one but less than fifteen amino acids.

3. The method of claim 1, wherein the peptide compound contains at least one but less than seven amino acids.

4. The method of claim 1, wherein the peptide compound is a dipeptide compound.

5. The method of claim 4, wherein the peptide compound is selected from the group consisting of Ri Asp GIy R₂; Ri Asp Ala R₂; Ri GIu GIy R₂; and Ri, GIu Ala R₂; wherein Ri is absent or is an amino terminal capping group, and R₂ is absent or is a carboxy terminal capping group.

6. The method of claim 5, wherein Ri is selected from the group consisting of a lipoic acid moiety (Lip); a glucose-3-O-glycolic acid moiety (Gga); 1 to 6 lysine residues; 1 to 6 arginine residues; a combination of 2 to 6 lysine and arginine residues; a thyronine group; an acyl group; and any combinations thereof.

7. The method of claim 1, wherein the peptide compound is represented by the following formula: Ri Asp GIy Xaa₃ Xaa₄ Xaa₅ R₂, wherein Rj is absent or is an amino terminal capping group; Xaa₃ is GIu or Leu; Xaa₄ is Ala or GIu, Xaa₅ is absent, Leu, or Ala, and R₂ is absent or is a carboxy terminal capping group.

8. The method of claim 1, wherein the peptide compound is selected from the group consisting of:

R₁ Asp GIy GIu Ala R₂ (SEQ ID NO: 14);

Ri, Asp GIy GIu Ala Leu R₂, (SEQ ID NO: 15);

Ri, Asp GIy Leu GIu Ala R₂, (SEQ ID NO: 16);

Ri, Asn Ala R₂;

Ri, Asn GIy R₂;

Ri, GIn GIyR₂;

Ri GIn Ala R₂;

R₁, Thr VaI Ser R₂;

Ri, Asp GIy Asp R₂; and

R₁ Asn GIy Asn R₂, wherein Ri, is absent or is an amino terminal capping group, and R₂ is absent or is a carboxy terminal capping group.

9. The method of claim 1, wherein the peptide compound has the formula:

Ri Xaai Xaa₂ Asp GIy Xaa₅ Xaa₆ Xaa₇ Xaa₈ Xaag Xaai₀ Xaaπ R₂, wherein R₁ is absent or is an amino terminal capping group; Xaai is absent or any amino acid; Xaa₂ is absent or any amino acid; Xaa₅ is GIu or Leu; Xaa₆ is Ala or GIu; Xaa₇ is absent, Leu, or Ala; Xaa₈ is absent or is any amino acid; Xaa₉ is absent or is any amino acid; Xaaio is absent or is any amino acid; Xaaπ is absent or is any amino acid; and R₂ is absent or is a carboxy terminal capping group.

10. The method of claim 1 , wherein the peptide compound has the formula:

R₁ Xaai Xaa₂ Xaa₃ R₂, wherein R₁ is absent or is an amino terminal capping group; Xaai is Asp, Asn, GIu, GIn, Thr, or Tyr; Xaa₂ is absent or is any amino acid; Xaa₃ is absent or is Asp, Asn, GIu, Thr, Ser, GIy, or Leu; and R₂ is absent or is a carboxy terminal capping group of the peptide compound.

11. The method of claims 1, wherein the peptide compound is administered in conjunction with another drug or with compounds useful for treating or preventing pulmonary inflammation.

12. The method of claim 1, wherein the peptide compound is selected from the group consisting of:

Lip Asp GIy;

Lip Asp Ala;

Lip GIu GIy;

Lip GIu Ala;

Ac Asp GIy;

Ac Asp Ala;

Ac GIu GIy;

Ac GIu Ala; T₃ Asp GIy;

T₃ Asp Ala;

T₃ GIu GIy;

T₃ GIu Ala;

Ac T₃ Asp GIy;

Ac T₃ Asp Ala;

Ac T₃ GIu GIy;

Ac T₃ GIu Ala;

Lip T₃ Asp GIy;

Lip T₃ Asp Ala;

Lip T₃ GIu GIy; and

Lip T₃ GIu Ala.

13. A method of treating or preventing chronic obstructive pulmonary disease in a subject in need thereof, comprising administering to the subject a composition comprising an effective amount of at least one peptide compound containing at least one but less than twenty amino acids.

14. The method of claim 13, wherein the chronic obstructive pulmonary disease is chronic bronchitis.

15. The method of claim 13, wherein the chronic obstructive pulmonary disease is emphysema.

16. A method of preparing a pharmaceutical composition for treating or preventing pulmonary inflammation in a subject in need thereof, comprising mixing a therapeutically effective amount of at least one peptide compound and a pharmaceutically acceptable carrier, wherein the peptide compound contains at least one but less than twenty amino acids.

17. A pharmaceutical composition prepared according to the method of claim 16.

18. A pharmaceutical composition comprising a therapeutically effective amount of at least one peptide compound and a pharmaceutically acceptable carrier, wherein the peptide compound has at least one but less than twenty amino acids.