Mucin hypersecretion inhibitors and methods of application thereof

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to peptides inhibiting mucin hypersecretion. The peptides have an amino acid sequence containing up to 24 amino acid residues of the sequence GAQFSKTAAKGEAAAERPGEAAVA which can have at least one amino acid substitute in said sequence selected from a group consisting of the substitute of A by K, the substitute of F, K, G, Q, S, T and/or E for A; or the substitute of Q for E.

EFFECT: preparation of a pharmaceutical composition on the basis of the peptides for mucin hypersecretion inhibition.

28 cl, 9 tbl, 5 ex

 

The technical field

The invention mainly relates to compositions comprising the peptides, and methods of use thereof.

The level of technology

Mucus is a biological fluid, capable of forming gels. It is a mixture of components, including water and secretions from the various cells. The mucines, also called glycoproteins of mucus or epithelial glycoproteins, are the major component of mucus and are glycoconjugates, which are characterized by numerous oligosaccharide side chains linked to the backbone of the peptide through the N - and O-linkages. Hypersecretion of mucin (a glycoprotein component of mucus) is observed in some respiratory diseases, including asthma, chronic bronchitis and cystic fibrosis (CF), and is a risk factor for death in patients with these diseases.

Airway the mucines released to the surface of the respiratory tract from the cells Goblet on the surface of the epithelium and cells of the submucosal mucus glands. The total number of surface fluid (mucus) in the respiratory tract is the result of speed mucus secretion in combination with the speed of its removal (by epithelial reabsorption, evaporation, ciliary transport and cough transport), i.e. the differences between the near the capacity of mucus secretion and the rate of its removal. Under "normal" conditions, the rate of secretion and removal of mucus is balanced so that the tracheobronchial tree is covered with only a thin surface layer of the liquid. Mucus hypersecretion (if it is not accompanied by a simultaneous increase in the uninstall process of mucus) leads to an overall increase in the number of mucus in normal conditions, which leads to the accumulation of mucus in the respiratory tract, which may in turn lead to airway obstruction and increased latency of the inhaled particles and microbial components.

Mucus hypersecretion contributes to the pathogenesis of a large number of inflammatory diseases of the respiratory tract in humans as well as animals other than humans. Increased secretion of mucus is manifested in chronic diseases such as asthma, chronic obstructive pulmonary disease (COPD) and chronic bronchitis; in genetic diseases such as cystic fibrosis; allergic States (atopy, allergic inflammation); if bronchiectasia; and in some acute infectious respiratory diseases such as pneumonia, rhinitis, flu and cold.

In many of these respiratory diseases hypersecretion of mucus is accompanied by an increased presence of airway inflammatory cells. These cells contribute to the atology of these diseases through the damage and destruction of tissues, produced by inflammatory mediators released from these cells. One example of such destruction by chronic inflammation observed in patients with cystic fibrosis, where released from neutrophil mediators (e.g. myeloperoxidase induce desquamation of epithelial tissues of the respiratory tract.

The respiratory tract of mammals covered by the lines of a thin layer of mucus produced and Sekretareva epithelial respiratory cells (cells Goblet) and submucosal glands. For diseases such as asthma, COPD, chronic bronchitis and cystic fibrosis, mucus hypersecretion is a common lesion. Excess mucus may contribute to the obstruction, susceptibility to infection and even the destruction of the walls of the respiratory tract and the surrounding tissue. The major components of mucus are satinowye glycoproteins that are synthesized secretory cells (i.e. cells Goblet cells and mucus) and accumulated inside the granules associated with the cytoplasmic membrane. The mucines are a family of glycoproteins that are secreted by epithelial cells, including cells of the respiratory, gastrointestinal and female reproductive tracts. The mucines responsible for the viscoelastic properties of mucus, and know of at least eight genes Muci the and. Cm. Patent application U.S. 10/180,753 (Publication No. U.S. 2003/0013652). Mucociliary damage caused by hypersecretion of mucin and/or hyperplasia of mucus cells leads to blockage of the respiratory tract mucus, which contributes to chronic infection, airway obstruction, and sometimes death. Many diseases of the Airways such as chronic bronchitis, chronic obstructive pulmonary disease, bronchiectasis, asthma, cystic fibrosis and bacterial infection, characterized by overproductive mucin. Cm. patent application U.S. No. 10/180,753 (Publication No. U.S. 2003/0013652). When appropriate stimulation satinowye granules are released by a process of exocytosis, in which the granules translucida to the periphery of the cells, where membrane granules fuse with the plasma membrane, leading to the secretion of the contents into the lumen.

Despite the obvious pathophysiological importance of this process, intracellular signaling mechanisms linking stimulation on the cell surface, releasing Musinovich granules were identified only recently. Cm. Li et al.,Journal of Biological Chemistry, 276: 40982-40990 (2001). I believe that monitorowanie rich in alanine protein substrate-kinase (MARCKS) is required for the secretion of mucus bronchial epithelial cells. Suggested that MARCKS binds on different sites with membrane and secretory granules and with the actin cytoskeleton, to serve as a physical connection between the reduction of the cytoskeleton and musinovym granules, and may play a role in the direction of secretory granules to the binding sites on the cell membrane. Cm. Singer et al., “A MARCKS-related peptide blocks mucus hypersecretion in a mouse model of asthma”,Nature Medicine, 10: 193-196 (2004). MANS peptide (myr-peptide 1) is monitorowania N-terminal amino acid sequence of 24 amino acids of the protein, called "Monitorowanie rich in alanine substrate With a kinase, which is usually referred to as MARCKS protein. It is shown that a fragment of MARCKS from 24 amino acids, monitorowania N-terminal sequence of the peptide (MANS), inhibits release of mucin in vitro and blocks mucus hypersecretion in a mouse model of asthma. Cm. Li et al. and Singer et al., above.

Known the importance of monitorowania to facilitate the translocation of peptides across membranes through lipid bilayer. A recent study demonstrated this importance, showing that demeritorious peptides do not pass through the cell membrane compared to monitorowanie peptides. Cm. A.Harishchandran et al., “Interaction of a Pseudosubstrate Peptide of Protein Kinase C and its Myristoylated Form with Lipid Vesicles. Only the Myzistoylated Form Translocates into Lipid Bilayer.”Biochem. Biophys. Acta, 1713: 73-82 (2005).

The invention

In one aspect, presents a peptide which is less than 24 am is nakilat and has the amino acid sequence, selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference amino acid sequence as PEPTIDE 1, which is also known as the MANS peptide, and (b) amino acid sequence essentially identical to the amino acid sequence defined in (a). One or more amino acids of the peptide optionally independently chemically modified, and the peptide has a mucin-inhibiting effect when administered to a mammal in a mucin-inhibiting amount.

In another aspect, presents a peptide which is less than 24 amino acids and has an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference amino acid sequence as the PEPTIDE 1; and (b) amino acid sequence essentially identical to the sequence defined in (a). N-terminal and C-terminal amino acid of the peptide is optionally independently chemically modified. The peptide has a mucin-inhibiting effect when administered to a mammal in a mucin-inhibiting amount and has a greater mucin-inhibiting effect in the mammal than the MANS peptide, with the introduction in equal concentrations.

In the following aspect presents a peptide which is less than 24 amino acids and has an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference amino acid sequence as the PEPTIDE 1; and (b) amino acid sequence essentially identical to the amino acid sequence defined in (a). N-terminal and C-terminal amino acid of the peptide is optionally independently chemically modified. The peptide has a greater solubility in water than the MANS peptide, and has a mucin-inhibiting effect when administered to a mammal in a mucin-inhibiting amount.

In another aspect, presents a method of inhibiting mucin hypersecretion in a mammal. The method includes the introduction of peptide mammal in a mucin-inhibiting amount that inhibits the secretion of mucin. The peptide consists of less than 24 amino acids and has an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference amino acid sequence as the PEPTIDE 1; and (b) amino acid sequence essentially identical to the sequence of ODA is divided in (a). One or more amino acids of the peptide optionally independently chemically modified.

In the following aspect presents a method of inhibiting mucin hypersecretion in a mammal. The method includes the introduction of a peptide to a mammal in a mucin-inhibiting amount that inhibits the secretion of mucin. The peptide consists of less than 24 amino acids and has an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference amino acid sequence as the PEPTIDE 1; and (b) amino acid sequence essentially identical to the sequence defined in (a). N-terminal and C-terminal amino acid of the peptide is optionally independently chemically modified, and the peptide has a greater mucin-inhibiting effect in the mammal than the MANS peptide, with the introduction in equal concentrations.

In the following aspect presents a method of inhibiting mucin hypersecretion in a mammal. The method includes the introduction of a peptide to a mammal in a mucin-inhibiting amount that inhibits the secretion of mucin. The peptide consists of less than 24 amino acids and has an amino acid sequence selected from the group consisting of: (a) amino acid sequence, Meuse is from 4 to 23 consecutive amino acids of the reference amino acid sequence, defined as PEPTIDE 1; and (b) amino acid sequence essentially identical to the sequence defined in (a). N-terminal and C-terminal amino acid of the peptide is optionally independently chemically modified, and the peptide has a greater solubility in water than the MANS peptide.

In another aspect, presents a peptide which is less than 24 amino acids and has the amino acid sequence of the variant amino acid sequence having from 4 to 23 consecutive amino acids of the reference amino acid sequence as the PEPTIDE 1. N-terminal and C-terminal amino acids of the peptide optionally chemically modified. The peptide has a mucin-inhibiting effect when administered to a mammal in a mucin-inhibiting amount has greater solubility in water than the MANS peptide, and has a greater mucin-inhibiting effect in the mammal than the MANS peptide, with the introduction in equal concentrations.

The peptides of the present invention is used for reducing and/or inhibiting mucin hypersecretion (i.e. decrease to normal or lower than normal levels) in the treatment of diseases and in the treatment of symptoms of diseases in which there is hypersecretion of mucin, as it is seen in States of chronic diseases, such as what STMA, chronic obstructive pulmonary disease (COPD) and chronic bronchitis; in genetic diseases such as cystic fibrosis; allergic States (atopy, allergic inflammation); if bronchiectasia; and in some acute infectious respiratory diseases such as pneumonia, rhinitis, flu and cold.

In the following embodiment presents the peptide consists of a sequence selected from the group consisting of: (a) amino acid sequences that have sequence GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1); and (b) amino acid sequence essentially identical to the sequence defined in (a); in which N-terminal amino acid of the peptide is not monitorowania and C-terminal amino acid of the peptide is optionally independently chemically modified, and the peptide has the effect of inhibiting mucin hypersecretion when administered to a mammal in an amount which inhibits mucin hypersecretion. This peptide is used for the treatment of hypersecretion of mucin in lung diseases.

Detailed description of preferred embodiments of the invention

The present invention relates to methods and compositions designed for a variety of purposes, including inhibition of mucin hypersecretion (i.e. inhibition of release of mucin) and Engibarov is the production of mucus (sometimes referred to here, as inhibition of mucus secretion) in a mammal. However, before describing further details of this invention, first you will define the following terms.

Definition

"Mucin-inhibiting effect, mucin-inhibiting activity or inhibition of the secretion of mucin" means reducing the amount of secretion of mucin (i.e. release of mucin) and does not necessarily mean the complete cessation of secretion of mucin. The introduction of the composition, with the mucin-inhibiting effect, leads to reduced secretion of mucin compared with that which could be observed or would be expected in the absence of such compositions. In one aspect, the amount of decrease in the secretion of mucin may be from about 5% number, which is secreted or hypersecretory above normal levels, up to about 100% of a quantity that is secreted or hypersecretory higher than normal levels of secretion. In another aspect, the amount of decrease in the secretion of mucin may be from about 5% number, which is secreted or hypersecretory above normal levels (i.e. about 5% of the amount greater above normal levels), up to the quantity that is below the normal level of secretion, such as about 50% of the amount secreted by normal levels of secretion.

"Goo-inhibiting the function effectively", "goo-inhibiting activity or inhibition of production of mucus" means the reduction of the production of mucus and does not necessarily mean the complete cessation of production of mucus. Introduction composition, with mucus-inhibiting effect, leads to a decrease in the production of mucus in comparison with what could be or might be expected in the absence of such a composition.

"Mucin-inhibiting amount" of a composition is an amount that reduces or inhibits the secretion of mucin (i.e. the release of mucin) compared with that which could be observed or would be expected in the absence of such compositions, such amount, which reduces the secretion of mucin from about 5% to about 100% of the amount of mucin, which hypersecretory above normal levels.

"Goo-inhibitory amount" of a composition is an amount that reduces or inhibits the production of mucus in comparison with what could be observed in the absence of such a composition.

In the reference peptide GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1) in the N-terminal position of the reference peptide G is in position 1; A in position 2 is adjacent to G in position 1; Q at position 3 is adjacent to A in position 2; F at position 4 is adjacent to Q at position 3; S in position 5 is adjacent to F in position 4; K is in position 6 at eget to S at position 5; T at position 7 is adjacent to K at position 6; A at position 8 is adjacent to T in position 7; A at position 9 is adjacent to A at position 8; K at position 10 is adjacent to A at position 9; G at position 11 is adjacent to K at position 10; E at position 12 is adjacent to G at position 11; A at position 13 is adjacent to E in position 12; A at position 14 is adjacent to A in position 13; A at position 15 is adjacent to A in position 14; E at position 16 is adjacent to A in position 15; R at position 17 is adjacent to E in position 16; P at position 18 is adjacent to R in position 17; G at position 19 is adjacent to P in position 18; E at position 20 is adjacent to G at position 19; A at position 21 is adjacent to E in position 20; A at position 22 is adjacent to A at position 21; V at position 23 is adjacent to A at position 22; A at position 24 is adjacent to V at position 23, in which the position of the 24 C-terminal position of the reference peptide.

"Variant" of a reference peptide or variant 4-23 amino acid segment of the reference peptide is a peptide that has an amino acid sequence that differs from the amino acid sequence of the reference peptide or amino acid sequence segment of the reference peptide, respectively, in at least one position of the reference amino acid of the peptide or amino acid sequence segment of the reference peptide, sootvetstvenno is, but which retains the mucin - or mucus-inhibitory activity, which usually varies between 0.1 and 10 times the activity of the reference peptide or segment, respectively, preferably between 0.2 and 6 times the activity of the reference peptide or segment, respectively, more preferably between 0.3 and 5-fold the activity of the reference peptide or segment, respectively. "Variant" of the reference amino acid sequence or variant 4-23 amino acid segment of the reference amino acid sequence is an amino acid sequence which differs by at least one amino acid from the reference amino acid sequence or segment of the reference amino acid sequence, respectively, but has the amino acid sequence of a peptide which retains the mucin - or mucus-inhibitory activity of the peptide or segment, respectively, encoded by the reference amino acid sequence, the activity of which usually varies between 0.1 and 10 times the activity of the peptide or segment of the reference sequence, respectively, preferably between 0.2 and 6 times the activity of the peptide or segment of the reference sequence, accordingly, more preferably between 0.3 and 5-fold activity of the peptide or segmentational sequence, respectively. Variant peptides with substitutions of amino acids or amino acid sequences with substitutions of amino acids may vary (i.e. different) from the reference peptide or a reference amino acid sequence one or more amino acid substitutions in the reference amino acid sequence; deletion variant peptide or deletion variant amino acid sequence may vary (i.e. different) from the reference peptide or a reference amino acid sequence one or more amino acid deletions in the reference amino acid sequence; and variant peptide with insertions of amino acids or amino acid sequences with insertions of amino acids may vary (i.e. different) from the reference sequence peptide or a reference amino acid sequence one or more amino acid insertions in the reference sequence. Variant peptide or variant amino acid sequences may be obtained by replacing one or more amino acids (for example, replacement of at least, 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) in the reference sequence, or may be obtained by deletion of one or more amino acids (e.g., deletions, at least, 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) in the reference members is Telenesti, or can be obtained by inserting one or more amino acids (for example, inserting at least, 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) in the reference sequence, or by combinations thereof in any order. Option 4-23 amino acid peptide segment with substitutions of amino acids or option 4-23 amino acid sequence segment with substitutions of amino acids may vary (i.e. different) from the reference 4-23 amino acid peptide segment or from the reference 4-23 amino acid sequence of a segment of one or more amino acid substitutions in the reference amino acid sequence segment; deletion option 4-23 amino acid peptide segment or 4-22 amino acid deletion variant amino acid sequence segment may vary (i.e. different) from 5-23 reference amino acid peptide segment or 5-23 reference amino acid sequence of a segment of one or more amino acid deletions in the reference amino acid sequence segment; and 4-23 variant amino acid peptide with insertions of amino acids or 4-23 amino acid variant amino acid sequence with insertions of amino acids may vary (i.e. different) from 4-22 reference amino acid sequence of the peptide or 4-22 reference amino acid aminoxy the pilot sequence one or more amino acid insertions in the reference sequence. 4-23 amino acid variant of the peptide or 4-23 amino acid variant amino acid sequences may be obtained by replacing one or more amino acids (for example, replacement of at least, 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) 4-23 amino acid segment of the reference amino acid sequence, or can be obtained by deletion of one or more amino acids (e.g., a deletion of at least, 1, 2, 3, 4, 5, 6, 7 or 8 amino acids), respectively, in most of the reference amino acid sequence, or may be obtained by inserting one or more amino acids (for example, inserting at least, 1, 2, 3, 4, 5, 6, 7 or 8 amino acids), respectively, in the smaller of the reference sequence, or by combinations thereof. Preferably the variant peptide or amino acid sequence differs from the reference peptide or segment of the reference peptide, or from the reference amino acid sequence, or from a segment of the reference amino acid sequence, respectively, less than 10 amino acid substitutions, deletions and/or insertions; more preferably less than 8 amino acid substitutions, deletions and/or insertions; even more preferably less than 6 amino acid substitutions, deletions and/or insertions; and even more preferably less than 5 amino acid substitutions, deletions and/or standing is to; and even more preferably less than 4 amino acid substitutions, deletions and/or insertions. Most preferably, the variant amino acid sequence differs from the reference peptide or segment of the amino acid sequence of one or two, or three amino acids.

"Sequence identity" with respect to amino acid sequences of the two peptides is the number of positions with identical amino acids divided by the number of amino acids in the shorter of the two sequences.

"Essentially identical" with respect to the comparison of amino acid sequences of two peptides or comparison of amino acid sequences of two peptide segments (e.g. segments of the reference peptide amino acid sequence) means that the amino acid sequence of peptides or segments of the peptide has at least 75% sequence identity, preferably at least 80% sequence identity, more preferably at least 90% sequence identity and most preferably at least 95% identity to the sequence.

The term "peptide"used here includes the peptide, as well as pharmaceutically acceptable salts of the peptide.

"Isolated" peptide, use the pot here means a natural peptide that separated or substantially separated from cellular components (e.g., nucleic acids and other peptides), which naturally accompany it, by purification, recombinant synthesis or by chemical synthesis, and includes not natural recombinante or chemically synthesized peptides that have been purified or substantially purified from cellular components, biological materials, chemical precursors and other chemicals.

The following three-letter and single-letter abbreviations of amino acids are used throughout the text: Alanine: (Ala); Arginine: (Arg) R, Asparagine: (Asn) N, Aspartic acid (Asp) D, Cysteine: (Cys), Glutamine: (Gln) Q, Glutamic acid (Glu, E, Glycine: (Gly) G Histidine: (His) H, Isoleucine: (Ile) I, Leucine: (Leu) L-Lysine: (Lys) K, Methionine: (Met) M, Phenylalanine: (Phe) F Proline: (Pro) P, serine: (Ser), Threonine: (Thr) T, Tryptophan: (Trp) W, Tyrosine: (Tyr Y Valine: (Val) V. Additional three-letter symbols of amino acids used here, include in parentheses (Hyp) for hydroxyproline (Nle) for norleucine, (Orn) for ornithine, (Pyr) for pyroglutamic acid (Sar) for sarcosine. Conventionally amino (or N-terminal end of the peptide is located at the left end of the written amino acid sequence of the peptide and the carboxyl (or C-terminal end is at the right end of the written amino acid is based sequence. Amino acid sequence of the peptide can be written in single-letter symbols to represent amino acids that are covalently bound to the peptide of the peptide amide bonds.

Table IX contains the list of amino acid sequences in the format of single-letter abbreviations, respectively, together with the corresponding peptide number and SEQ ID NO. The reference amino acid sequence of a peptide listed as peptide 1. Amino acid sequences of the peptides of the invention having the amino acid sequence of from 4 to 23 consecutive amino acids of the reference amino acid sequence listed as peptides 2-231 together with the amino acid sequence of random N-terminal sequence (RNS), including amino acids MANS peptide, designated as peptide 232. Amino acid sequence illustrative amino acid sequence variants of the peptides of the invention as described herein also listed as peptides 233-245 and 247-251. Assume that the listed variants of the peptides are not limiting group of peptides and presents only to serve as illustrative examples of peptides of the invention. Also presents illustrative reverse serial amino acid is inost and illustrative random amino acid sequence of peptide according to the invention. It is implied that shown in the table opposite and random amino acid sequences are not illustratively according to the invention.

Amino acid sequence of a peptide listed in the Table IX, may be chemically modified. For example, if the amino acid sequence of a peptide listed in the Table IX, chemically modified at the N-terminal amino groups with the formation of amide carboxylic acid, the resulting peptide is sometimes referred here by a combination of the identifier of the carboxylic acid represented by the console, which is connected by a hyphen with a peptide room. For example, in relation to peptide 79 as an example, the N-terminal monitorowanie peptide 79 can sometimes be described as "monitorowanie peptide 79 or myr-peptide 79", N-terminal acetylated peptide 79 can sometimes be described as "acetyl-peptide 79" or "Ac-peptide 79". Cyclic variant of the peptide 79 may be designated as "cyclic peptide 79" or "cyc-peptide 79". Also, for example, if the amino acid sequence of a peptide listed in the Table IX, chemically modified C-terminal carboxyl group, for example, with an amine, such as ammonia, with the formation of the C-terminal amide, the resulting peptide is sometimes referred here by a combination of the identifier OST the TKA amine, represented by a suffix, which is connected by a hyphen with the number of peptide. Thus, for example, C-terminal amide peptide 79 can sometimes be described as "peptide-NH2". When the N-terminal amino group of the peptide (e.g., peptide 79) chemically modified, for example, myristoleic group and the C-terminal carboxyl group is chemically modified, for example, ammonium group, amide formation as described above, the resulting peptide can sometimes be denoted using symbols and prefixes, and suffixes, such as "myr-peptide 79-NH2".

The invention includes peptides having amino acid sequences comprising less than 24 amino acids from the amino acid sequences of related amino acid sequences MANS peptide (i.e. MANS peptide is myristoyl-PEPTIDE 1 and reference 24-amino acid sequence of MANS peptide is a PEPTIDE of 1). The peptides of this invention consist of amino acid sequences containing less than 24 amino acids, and may consist of: 8-14, 10-12, 9-14, 9-13, 10-13, 10-14, at least 9, at least 10, or from a similar number of amino acids. Peptides are usually unbranched chain, but can also be cyclic peptides. In addition, peptides can be selected peptides is.

In relation to PEPTIDE 1, reference 24 amino acid sequence segment of 23 consecutive amino acids of the reference amino acid sequence, sometimes referred to here as a 23-Mer. Similarly, the segment of 22 consecutive amino acids of the reference sequence, sometimes referred to here as a 22-Mer; a sequence of 21 amino acids as a 21-Mer; the sequence of the 20 amino acids as 20-Mer; a sequence of 19 amino acids as a 19-Mer; a sequence of 18 amino acids as an 18-Mer; a sequence of 17 amino acids as a 17-Mer; a sequence of 16 amino acids as 16-Mer; a sequence of 15 amino acids as 15-Mer; a sequence of 14 amino acids as a 14-Mer; a sequence of 13 amino acids as a 13-Mer; the sequence of the 12 amino acids as a 12-Mer; a sequence of 11 amino acids as 11-Mer; a sequence of 10 amino acids as 10-Mer; a sequence of 9 amino acids with 9-Mer; a sequence of 8 amino acids as an 8-Mer; a sequence of 7 amino acids such as 7-Mer; a sequence of 6 amino acids as 6-Mer; a sequence of 5 amino acids as 5-Mer; and a sequence of 4 amino acids as 4-Mer. In one aspect of any of these amino acid sequences, labeled "4-Mer-23-Mer", which are themselves peptide and (sometimes marked here as H 2N-peptide-COOH)may be independently chemically modified, for example, chemical modification and chemical modification can be selected from the group consisting of: (i) formation of amide N-terminal amino group (H2N-peptide-), such as, for example, preferably C1 or C2 (acetic acid) and to C22 carboxylic acid; (ii) formation of amide at the C-terminal carboxyl group (-peptide-COOH), such as, for example, with ammonia or with C1-C22 primary or secondary amine; and (iii) combinations thereof.

The peptides have an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence PEPTIDE 1; (b) a sequence essentially similar to the amino acid sequence defined in (a); and (c) the variant amino acid sequence defined in (a)and variant selected from the group consisting of variants with substitutions of amino acids, deletion variants, variants with insertions of amino acids and their combinations. In some embodiments, the peptides have an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 8 to 14 consecutive amino acids of the reference sequence PEPTIDE 1; (b) aminoxy the pilot sequence, essentially identical to the sequence defined in (a); and (c) the variant amino acid sequence defined in (a)and variant selected from the group consisting of variants with substitutions of amino acids, deletion variants, variants with insertions of amino acids and their combinations. In yet another embodiment, the peptides have an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 10 to 12 consecutive amino acids of the reference sequence PEPTIDE 1; (b) amino acid sequence essentially identical to the sequence defined in (a); and (c) the variant amino acid sequence defined in (a)and variant selected from the group consisting of variants with substitutions of amino acids, deletion variants, variants with insertions of amino acids and their combinations. In further embodiments, the peptides have an amino acid sequence having at least 9, at least 10, 9-14, 9-13, 10-13, 10-14, or such number of consecutive amino acids of the reference sequence PEPTIDE 1; amino acid sequence essentially they are identical; or their variant and variant selected from the group consisting of variants with substitutions of amino acids, deletion variants, variants with insertions linakis is from, and combinations thereof. As explained further below, one or more amino acids, peptides (for example, N-terminal and/or C-terminal amino acids) may be optionally independently chemically modified; in some embodiments, one or more amino acids of the peptide will be chemically modified, while in other embodiments, none of the amino acids of the peptide is not chemically modified. In one aspect of the preferred modification may occur at the amino (H2N) the group of N-terminal amino acid of the peptide or peptide segment (and the amino group forms a peptide amide bond, if present within the peptide sequence, and not in the N-terminal position). In another preferred aspect, the modification can occur in the carboxyl (-COOH) group of C-terminal amino acids of the peptide or peptide segment (and the carboxyl group forms a peptide amide bond, if present within the peptide sequence, and not in the C-terminal position). In another preferred aspect, the modification can occur in the N-terminal amino (H2N-) group, and the C-terminal carboxyl (-COOH) group.

In some embodiments, the amino acid sequence of the peptide begins with the N-terminal amino acids of the reference sequence PEPTIDE 1. For example, the peptide can have the amino acid on sledovatelnot, selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence PEPTIDE 1, which begins with the N-terminal amino acids of the reference sequence (i.e., the PEPTIDE 2, PEPTIDE 4, PEPTIDE 7 PEPTIDE 11, PEPTIDE 16, PEPTIDE 22, PEPTIDE 29, 37 PEPTIDE, PEPTIDE 46, PEPTIDE 56, PEPTIDE 67, PEPTIDE 79, PEPTIDE 92, PEPTIDE 106, PEPTIDE 121, PEPTIDE 137, 154 PEPTIDE, PEPTIDE 172, PEPTIDE 191 or PEPTIDE 211); (b) amino acid sequences essentially similar to the amino acid sequence defined in (a); and (c) the variant amino acid sequence defined in (a).

In other embodiments, the amino acid sequence of the peptide ends C-terminal amino acid of the reference sequence PEPTIDE 1. For example, the peptide can have the amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence PEPTIDE 1, which ends the C-terminal amino acid of the reference sequence (i.e., the PEPTIDE 3, PEPTIDE 6, PEPTIDE 10, PEPTIDE 15, PEPTIDE 21, PEPTIDE 28, PEPTIDE 36, PEPTIDE 45, 55 PEPTIDE, PEPTIDE 66, PEPTIDE 78, PEPTIDE 91, PEPTIDE 105, PEPTIDE 120, PEPTIDE 136, PEPTIDE 153, PEPTIDE 171, PEPTIDE 190, PEPTIDE 210 and PEPTIDE 231); (b) amino acid p is coherence, essentially similar to the amino acid sequence defined in (a); and (c) the variant amino acid sequence defined in (a).

In other embodiments, the amino acid sequence of the peptide does not begin with N-terminal amino acids of the reference sequence PEPTIDE 1, and often begins with the amino acids in position 2 and to the amino acid at position 21 of the reference sequence PEPTIDE 1. For example, the peptide can have the amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence PEPTIDE 1, which begins with any of the amino acids between positions 2 and 21 of the reference sequence. These peptides can be of length from 4 to 23 consecutive amino acids and can present peptides in the middle of the reference sequence PEPTIDE 1; (b) amino acid sequences essentially similar to the amino acid sequence defined in (a); and (c) the variant amino acid sequence defined in (a). These peptides disclosed in Table IX.

Peptide amino acid sequences, which are used in this invention in order to inhibit mucin hypersecretion in a mammal, and which is used to reduce the IC number of mucin hypersecretion in a mammal, and used in methods of inhibiting mucin hypersecretion and ways of reducing hypersecretion of mucin, include amino acid sequences of the selected peptides and amino acid sequences of the peptides of the present invention that do not contain N-terminal and/or C-terminal chemically modified group, and the peptide amino acid sequence selected from the group consisting of a 23-Mer (i.e. peptides having a sequence of 23 amino acids): PEPTIDE 2; and PEPTIDE 3; 22-mers (i.e. peptides having a sequence of 22 amino acids): PEPTIDE 4; PEPTIDE 5; and PEPTIDE 6; 21-Mer (i.e. peptides having a sequence of 21 amino acids): PEPTIDE 7; PEPTIDE 8; PEPTIDE 9; and PEPTIDE 10; 20-mers (i.e. peptides having the sequence of 20 amino acids): PEPTIDE 11; PEPTIDE 12; PEPTIDE 13; PEPTIDE 14; and PEPTIDE 15; 19-Mer (i.e. peptides having a sequence of 19 amino acids): PEPTIDE 16; PEPTIDE 17; PEPTIDE 18; PEPTIDE 19; PEPTIDE 20; and PEPTIDE 21; 18-mers (i.e. peptides having a sequence of 18 amino acids): PEPTIDE 22; PEPTIDE 23; peptide 25; peptide 26; peptide 27; and peptide 28; 17-mers (i.e. peptides having a sequence of 17 amino acids): peptide 29; peptide 30; peptide 31; peptide 32; peptide 33; peptide 34; peptide 35; and peptide 36; 16-mers (i.e. peptides having a sequence of 16 amino acids): peptide 37; PE is Ted 38; peptide 39; 40 peptide; peptide 41; 42 peptide; peptide 43; 44 peptide; peptide 45; 15-mers (i.e. peptides having a sequence of 15 amino acids): peptide 46; peptide 47; peptide 48; peptide 49; peptide 50; peptide 51; peptide 52; peptide 53; peptide 54; and 55 peptide; 14-mers (i.e. peptides having a sequence of 14 amino acids): peptide 56; 57 peptide; peptide 58; peptide 59; peptide 60; peptide 61; peptide 62; 63 peptide; peptide 64; peptide 65; and peptide 66; 13-mers (i.e. peptides having a sequence of 13 amino acids): peptide 67; peptide 68; peptide 69; peptide 70; peptide 71; peptide 72; peptide 73; peptide 74; peptide 75; peptide 76; peptide 77; and peptide 78; 12-mers (i.e. peptides having a sequence of 12 amino acids): peptide 79; peptide 80; peptide 81; peptide 82; peptide 83; peptide 84; peptide 85; peptide 86; peptide 87; 88 peptide; peptide 89; 90 peptide; peptide 91; 11-mers (i.e. peptides having a sequence of 11 amino acids): peptide 92; peptide 93; peptide 94; peptide 95; peptide 96; peptide 97; peptide 98; 99 peptide; peptide 100; peptide 101; peptide 102; peptide 103; peptide 104; and peptide 105; 10-mers (i.e. peptides having a sequence of 10 amino acids): peptide 106; peptide 107; peptide 108; peptide 109; peptide 110; peptide 111; peptide 112; peptide 113; peptide 114; peptide 115; peptide 116; 117 peptide; peptide 118; 119 peptide; peptide 120; 9-mers (i.e. peptides having a sequence of 9 amino acids): peptide 121; peptide 122; peptide 123; peptide 124; peptide 125; Atid 126; peptide 127; peptide 128; peptide 129; peptide 130; peptide 131; peptide 132; peptide 133; peptide 134; 135 peptide; peptide 136; 8-mers (i.e. peptides having a sequence of 8 amino acids): peptide 137; peptide 138; peptide 139; 140 peptide; peptide 141; peptide 142; peptide 143; peptide 144; peptide 145; peptide 146; peptide 147; peptide 148; 149 peptide; peptide 150; 151 peptide; peptide 152; and peptide 153; 7-Mer (i.e. peptides having a sequence of 7 amino acids): peptide 154; 155 peptide; peptide 156; peptide 157; peptide 158; peptide 159; peptide 160; peptide 161; 162 peptide; peptide 163; peptide 164; peptide 165; peptide 166; 167 peptide; peptide 168; peptide 169; peptide 170; and peptide 171; 6-mers (i.e. peptides having a sequence of 6 amino acids): peptide 172; peptide 173; peptide 174; peptide 175;

peptide 176; peptide 177; peptide 178; peptide 179; peptide 180; peptide 181; peptide 182; peptide 183; peptide 184; peptide 185; peptide 186; peptide 187; peptide 188; peptide 189; and peptide 190; 5-mers (i.e. peptides having the sequence of 5 amino acids): peptide 191; peptide 192; peptide 193; peptide 194; peptide 195; peptide 196; peptide 197; peptide 198; peptide 199; peptide 200; peptide 201; peptide 202; peptide 203; peptide 204; peptide 205; peptide 206; peptide 207; peptide 208; peptide 209; and peptide 210; and 4-mers (i.e. peptides having a sequence of 4 amino acids): peptide 211; peptide 212; peptide 213; peptide 214; peptide 215; peptide 216; 217 peptide; peptide 218; peptide 219; 220 peptide; peptide 221; peptide 222; peptide 223; pepcid; peptide 225; peptide 226; 227 peptide; peptide 228; peptide 229; peptide 230; and peptide 231.

Preferred amino acid sequences of the selected peptides and chemically modified at the N-terminal and/or C-terminal group of the peptides of the present invention is selected from the group consisting of a 23-Mer: PEPTIDE 2; and PEPTIDE 3; 22-mers: PEPTIDE 4; PEPTIDE 5; and PEPTIDE 6; 21-mers: PEPTIDE 7; PEPTIDE 8; PEPTIDE 9; and PEPTIDE 10; 20-mers: PEPTIDE 11; PEPTIDE 12; PEPTIDE 13; PEPTIDE 14; and PEPTIDE 15; 19-mers: PEPTIDE 16; PEPTIDE 17; PEPTIDE 18; PEPTIDE 19; PEPTIDE 20; and PEPTIDE 21; 18-mers: PEPTIDE 22; PEPTIDE 23; peptide 24; peptide 25; peptide 26; peptide 27; and peptide 28; 17-mers: peptide 29; peptide 30; peptide 31; peptide 32; peptide 33; peptide 34; peptide 35; and peptide 36; 16-mers: peptide 37; peptide 38; peptide 39; 40 peptide; peptide 41; 42 peptide; peptide 43; 44 peptide; peptide 45; 15-mers: peptide 46; peptide 47; peptide 48; peptide 49; peptide 50; peptide 51; peptide 52; peptide 53; and the peptide 54; 14-mers: peptide 56; 57 peptide; peptide 58; peptide 59; peptide 60; peptide 61; 62 peptide; peptide 63; and peptide 64; 13-mers: peptide 67; peptide 68; peptide 69; peptide 70; peptide 71; peptide 72; peptide 73; peptide 74; and the peptide 75; 12-Mer: peptide 79; peptide 80; peptide 81; peptide 82; peptide 83; peptide 84; peptide 85; peptide 86; and peptide 87; 11-mers: peptide 92; peptide 93; peptide 94; peptide 95; peptide 96; peptide 97; peptide 98; 99 peptide; peptide 100; 10-mers: peptide 106; peptide 107; peptide 108; peptide 109; peptide 110; peptide 111; peptide 112; pepti the 113; and peptide 114; 9-mers: peptide 122; peptide 123; peptide 124; peptide 125; 126 peptide; peptide 127; peptide 128; and peptide 129; 8-mers: peptide 139; 140 peptide; peptide 141; peptide 142; peptide 143; peptide 144; and peptide 145; 7-mers: peptide 157; peptide 158; peptide 159; peptide 160; peptide 161; and peptide 162; 6-mers: peptide 176; peptide 177; peptide 178; peptide 179; and peptide 180; 5-mers: peptide 196; peptide 197; peptide 198; and peptide 199; and 4-mers: peptide 217; and peptide 219.

More preferred amino acid sequences of the selected peptides and chemically modified at the N-terminal and/or C-terminal group of the peptides of the present invention is selected from the group consisting of a 23-Mer: peptide 2; and peptide 3; 22-mers: peptide 4; peptide 5; and peptide 6; 21-mers: peptide 7; peptide 8; peptide 9; and peptide 10; 20-mers: peptide 11; peptide 12; peptide 13; peptide 14; and peptide 15; 19-mers: peptide 16; peptide 17; peptide 18; peptide 19; peptide 20; and peptide 21; 18-mers: peptide 22; peptide 23; peptide 24; peptide 25; peptide 26; peptide 27; and peptide 28; 17-mers: peptide 29; peptide 30; peptide 31; peptide 32; peptide 33; peptide 34; peptide 35; and peptide 36; 16-mers: peptide 37; peptide 38; peptide 39; 40 peptide; peptide 41; 42 peptide; peptide 43; 44 peptide; peptide 45; 15-mers: peptide 46; peptide 47; peptide 48; peptide 49; peptide 50; peptide 51; peptide 52; peptide 53; and the peptide 54; 14-mers: peptide 56; 57 peptide; peptide 58; peptide 59; peptide 60; peptide 61; 62 peptide; peptide 63; and peptide 64; 13-mers: peptide 67; peptide 68; peptide 69; peptide 70 peptide 71; peptide 72; peptide 73; peptide 74; peptide 80; peptide 81; peptide 82; peptide 83; peptide 84; peptide 85; peptide 86; and peptide 87; 11-mers: peptide 92; peptide 93; peptide 94; peptide 95; peptide 96; peptide 97; peptide 98; 99 peptide; peptide 100; 10-mers: peptide 106; peptide 108; peptide 109; peptide 110; peptide 111; peptide 112; peptide 113; and peptide 114; 9-mers: peptide 124; peptide 125; 126 peptide; peptide 127; peptide 128; and peptide 129; 8-mers: peptide 141; peptide 142; peptide 143; peptide 144; and peptide 145; 7-mers: peptide 159; peptide 160; peptide 161; and peptide 162; 6-mers: peptide 178; peptide 179; and peptide 180; 5-mers: peptide 198; and peptide 199; and 4-Mer: peptide 219.

In other embodiments, the amino acid sequence of the peptide comprises sequentially located the remains of A, K, G, and E as in the peptide 219 of the reference sequence PEPTIDE 1. For example, the peptide can have the amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence PEPTIDE 1, which includes consistently located the remains of A, K, G, and E as in the peptide 219 of the reference sequence PEPTIDE 1 (for example, PEPTIDE 219, PEPTIDE 45, PEPTIDE 79, PEPTIDE 67, PEPTIDE 80 and so on); (b) amino acid sequences essentially similar to the amino acid sequence defined in (a); and (c) the variant amino acid serial the major, defined in (a).

Examples of peptide segments that contain the amino acid sequence AKGE reference peptide amino acid sequence of PEPTIDE 1, include (a) 23-measures: peptide 2; and peptide 3; 22-measures: peptide 4; peptide 5; and peptide 6; 11-measures: peptide 7; peptide 8; peptide 9; and peptide 10; 20-measures: peptide 11; peptide 12; peptide 13; peptide 14; and peptide 15; 19-measures: peptide 16; peptide 17; peptide 18; peptide 19; peptide 20; and the peptide 21; 18-measures: peptide 22; peptide 23; peptide 24; peptide 25; peptide 26; peptide 27; and peptide 28; 17-measures: peptide 29; peptide 30; peptide 31; peptide 32; peptide 33; peptide 34; peptide 35; and peptide 36; 16-measures: 37 peptide; peptide 38; peptide 39; 40 peptide; peptide 41; 42 peptide; peptide 43; 44 peptide; peptide 45; 15-measures: peptide 46; peptide 47; peptide 48; peptide 49; peptide 50; peptide 51; peptide 52; peptide 53; and the peptide 54; 14-measures: peptide 56; 57 peptide; peptide 58; peptide 59; peptide 60; peptide 61; 62 peptide; peptide 63; and peptide 64; 13-measures: peptide 67; peptide 68; peptide 69; peptide 70; peptide 71; peptide 72; peptide 73; peptide 74; and the peptide 75; 12-measures: peptide 79; peptide 80; peptide 81; peptide 82; peptide 83; peptide 84; peptide 85; peptide 86; and peptide 87; 11-measures: peptide 93; peptide 94; peptide 95; peptide 96; peptide 97; peptide 98; 99 peptide; peptide 100; 10-measures: peptide 108; peptide 109; peptide 110; peptide 111; peptide 112; peptide 113; and peptide 114; 9-measures: peptide 124; peptide 125; 126 peptide; peptide 127; peptide 128; and peptide 129; 8-measures: peptide 141; peptide 142; peptide 143; peptide 144; and peptide 15; 7-measures: peptide 159; peptide 160; peptide 161; and peptide 162; 6-measures: peptide 178; peptide 179; and peptide 180; 5-measures: peptide 198; and peptide 199; and 4-Mer: peptide 219, (b) a sequence essentially similar to the amino acid sequence defined in (a); and (c) a variant amino acid sequence defined in (a)and variant selected from the group consisting of variants with substitutions of amino acids, deletion variants, variants with insertions amino acids and combinations thereof, where the segment comprises or consists of 4-23 consecutive amino acids.

In another preferred embodiment the peptide sequence has an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 10 to 23 consecutive amino acids of the reference sequence peptide 1; (b) a sequence essentially similar to the amino acid sequence defined in (a); and (c) the variant amino acid sequence defined in (a)and variant selected from the group consisting of variants with substitutions of amino acids, deletion variants, variants with insertions of amino acids and combinations thereof, where the preferred amino acid sequences include the 23-Mer: peptide 2; 22-Mer: peptide 4; 21-Mer: peptide 7; 20-Mer: peptide 11; 19-Mer: peptide 16; 18-Mer: peptide 22; 17-Mer: peptide the ID 29; 16-Mer: peptide 37; 15-Mer: peptide 46; 14-Mer: peptide 56; 13-Mer: peptide 67; 12-Mer: peptide 79; 11-Mer: peptide 92; and 10-Mer: peptide 106.

In further embodiments the amino acid sequence of the peptide begins with the N-terminal amino acids of the reference sequence PEPTIDE 1 and includes consistently located the remains of A, K, G, and E as in the peptide 219 of the reference sequence PEPTIDE 1, while in other embodiments the amino acid sequence of the peptide ends C-terminal amino acid of the reference sequence PEPTIDE 1 and includes consistently located the remains of A, K, G, and E as in the peptide 219 of the reference sequence PEPTIDE 1.

The peptides may include one or more amino acid deletions, substitutions and/or insertions relative to the reference amino acid sequence. Preferably, the substitutions can be conservative amino acid substitutions or substitutions can be conservative amino acid substitutions. In some embodiments the peptides, including peptides with amino acid sequences that are essentially identical to a reference amino acid sequence or the variant will not have deletions or insertions, compared with the corresponding successive amino acids of the reference amino acid sequence, but mo is ut to be conservative or not conservative replacement. Amino acid substitutions that can be made in the reference amino acid sequence of the peptides of the invention include, but are not limited to, the following substitutions: alanine (A) can be replaced by a lysine (K), valine (V), leucine (L) or isoleucine (I); glutamic acid (E) can be replaced by aspartic acid (D); glycine (G) may be replaced by Proline (P); lysine (K) may be replaced by arginine (R), glutamine (Q) or aspargine (N); phenylalanine (F) may be replaced by a leucine (L), valine (V), isoleucine (I) or alanine (A); Proline (P) can be replaced by a glycine (G); glutamine (Q) can be replaced by glutamic acid (E) or asparagine (N); arginine (R) may be replaced by lysine (K), glutamine (Q) or asparagine (N); serine (S) may be replaced by a threonine (T); threonine (T) can be replaced by a serine (S); valine (V) may be replaced by a leucine (L), isoleucine (I), methionine (M), phenylalanine (F), alanine (A) or norleucine (Nle). For example, substitutions can be made in the reference amino acid sequence of the peptides of the invention include the substitution of phenylalanine (F) by alanine (A) (for example, the amino acid at position 4 of the reference amino acid sequence), a substitution of glutamine (Q) to glutamic acid (E) (for example, the amino acid at position 3 of the reference amino acid sequence), a substitution of alanine (A lysine (K) (for example, the amino acid in position 2 and/or 8 of the reference amino acid sequence), and/or substitution of threonine (T) by serine (S) (e.g., the amino acid in position 7 of the reference amino acid sequence).

When substitutions include amino acid sequences of the peptides of the invention (and peptides include both modified and chemically modified peptides, for example, using N-terminal and/or C-terminal modifications, such as amide formation), in relation to a reference amino acid sequence, preferably at least 80% sequence identity between the amino acid sequence of the peptide and the reference amino acid sequence. Peptides having from 5 to 23 amino acids and comprising one amino acid substitution relative to a reference amino acid sequence will be from about 80% to about 96% (i.e. ~95,7%) sequence identity to the reference amino acid sequence. Peptides having from 10 to 23 amino acids and comprising one amino acid substitution relative to a reference amino acid sequence will be from about 90% to about 96% (i.e. ~95,7%) sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and includes one aminoxy the pilot replacement in relation to a reference amino acid sequence, will be from about 95% to about 96% (i.e. ~95,7%) sequence identity to the reference amino acid sequence. Peptides having from 10 to 23 amino acids and comprising two amino acid substitutions relative to a reference amino acid sequence will be from about 80% to about 92% (i.e. ~91.3 percent) sequence identity to the reference amino acid sequence. Peptides having from 16 to 23 amino acids and comprising two amino acid substitutions relative to a reference amino acid sequence will be from about 87,5% to about 92% (i.e. ~91.3 percent) sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and comprising two amino acid substitutions relative to a reference amino acid sequence will be from about 90% to about 92% (i.e. ~91.3 percent) sequence identity to the reference amino acid sequence. Peptides having from 15 to 23 amino acids and comprising three amino acid substitutions relative to a reference amino acid sequence will be from about 80% to about 87% sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and comprising three amino acid substitutions relative to et the reference amino acid sequence, will have from about 85% to about 87% sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and comprising four amino acid substitutions relative to a reference amino acid sequence will be from about 80% to about 83% (i.e. ~82,6%) sequence identity to the reference amino acid sequence.

In the peptides of this invention in relation to consistently located to the amino acid sequence of the reference peptide (which is 24-Merom) by the replacement of one amino acid in the sequence of 23 consecutive amino acids (23-Mer), selected from the reference sequence of 24 amino acids, receive a peptide with an amino acid sequence that has 95,65% (or ~96%) sequence identity with the amino acid segment in the reference peptide with which the 23-Mer has its own identity. Similarly, the replacement of two, three, four and five amino acids in the specified 23-Mer receive a peptide with an amino acid sequence that has 91,30% (or ~91%), 86,96% (or ~87%), 82,61% (or ~83%) and 78,27% (or ~78%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one, two, three, four and five amino acids in the 22nd floor as the given peptide with the amino acid sequence, which has 95,45% (~95%), 90,91% (or ~91%), 86,36% (or ~86%), 81,82% (or ~82%) and 77,27% (or ~77%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one, two, three, four and five amino acids in the 21-Mer receive a peptide with an amino acid sequence that has 95,24% (~95%), 90,48% (~91%), 85,71% (~86%), 80,95 (~81%) and 76,19% (~76%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one, two, three, four and five amino acids in the 20 least get to a peptide with an amino acid sequence that has 95,00% (95%), 90,00% (90%), 85,00% (85%), 80,00% (80%) and 75,00% (75%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, the replacement of one, two, three and four amino acids in the 19-Mer receive a peptide with an amino acid sequence that has 94,74% (~95%), 89,47% (~89%), 84,21% (~84%) and 78,95% (~78%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, the replacement of one, two, three and four amino acids in the 18-Mer receive a peptide with an amino acid sequence that has 94,44% (~94%), 88,89% (~89%), 83,33% (~83%) and 77,78% (~79%) sequence identity with the amino acid sequence of a reference peptide is, respectively. Similarly, the replacement of one, two, three and four amino acids in the 17-Mer receive a peptide with an amino acid sequence that has 94,12% (~94%), 88,23% (~88%), 82,35% (~82%) and 76,47% (~76%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, the replacement of one, two, three and four amino acid 16-Mer receive a peptide with an amino acid sequence that has 93,75% (~94%), 87,50% (~88%), 81,25% (~81%) and 75,00% (75%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one, two and three amino acids in the 15-Mer receive a peptide with an amino acid sequence that has 93,33% (~93%), 86,67% (~87%) and 80.00% (80%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one, two and three amino acids in the 14-Mer receive a peptide with an amino acid sequence that has 92,86% (~93%), 85,71% (~86%) and 78,57% (79%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one, two and three amino acids in the 13-Mer receive a peptide with an amino acid sequence that has 92,31% (~92%), 84,62% (~85%) and 76,92% (~77%) sequence identity with the amino acid pic what ecovitality reference peptide, respectively. Similarly, replacement of one, two and three amino acids in the 12-Mer receive a peptide with an amino acid sequence that has 91,67% (~92%), 83,33% (~83%) and 75,00% (75%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one or two amino acids in the 11-Mer receive a peptide with an amino acid sequence that has 90,91% (~91%) and 81,82% (~82%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one or two amino acids in the 10-Mer receive a peptide with an amino acid sequence that has 90,00% (90%) 80,00% (80%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one or two amino acids in the 9-Mer receive a peptide with an amino acid sequence that has 88,89% (~89%) and 77,78% (~78%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, replacement of one or two amino acids in the 8-Mer receive a peptide with an amino acid sequence that has 87,50% (~88%) and 75,00% (75%) sequence identity with the amino acid sequence of the reference peptide, respectively. Similarly, the replacement of one amino acid is you 7-least 6-Mer, 5-Mer and 4-Mer receive a peptide with an amino acid sequence that has 85,71% (~86%), 83,33% (~83%), 80,00% (80%) and 75,00% (75%) sequence identity with the amino acid sequence of the reference peptide, respectively. Preferred amino acid sequences of this invention have more than 80% sequence identity with the amino acid sequence of the reference sequence, more preferably, if the sequence identity with the amino acid sequence of the reference sequence will be 81%-96%, and more preferably, if the sequence identity with the amino acid sequence of the reference sequence will be 80%-96%. Preferred amino acid sequences may not necessarily be chemically bound N-terminal amino group of the peptide amide bond with a linear component With2-C22aliphatic carboxylic acid, more preferably a linear component With2-C16aliphatic carboxylic acid, most preferably a linear component With2or16aliphatic carboxylic acids and optionally chemically bound in the C-terminal carboxyl group of the peptide amide bond with the amino group, such as ammonia or primary or secondary s is nagrampa, such as linear C1-C16aliphatic primary amino group.

Examples of peptide with the substitution of amino acids 79, 12-measure, include, for example peptide 238, where Q at position 3 of the peptide 79 replaced by E in the sequence 238; peptide 233, where A is in position 2 of the peptide 79 replaced by K in the peptide 233; peptide 234, where A in position 8 of the peptide 79 replaced by K in the peptide 234; peptide 235, where A at positions 2 and 8 of the peptide 79 replaced by K in the peptide 235; peptide 237, where F in position 4 of the peptide 79 was replaced by A peptide 237; peptide 239, where K in position 10 peptide 79 was replaced by A in peptide 239; peptide 240, where G in position 11 peptide 79 was replaced by A in peptide 240; and peptide 241, where E in position 12 peptide 79 was replaced by A in peptide 241.

Examples of peptide 106 replacing amino acids, 10-measure, include, for example peptide 236, where F in position 4 of the peptide 106 was replaced by A in peptide 236; peptide 242, where G in position 1 of the peptide 106 was replaced by A in peptide 242; 243 peptide, where Q at position 3 of the peptide 106 was replaced by A in peptide 243; peptide 244, where S is in position 5 of the peptide 106 was replaced by A in peptide 244; peptide 245, where K in position 6 peptide 106 was replaced by A in peptide 245; peptide 247, where T is in position 7 of the peptide 106 was replaced by A in peptide 247; peptide 248, where K in position 10 peptide 106 was replaced by A in peptide 248; peptide 249, where both K at positions 6 and 10 peptide 106 was replaced by A in peptide 249.

Examples options PE the Chida 137 replacing amino acids, 8-measure, include, for example peptide 250, where F in position 4 of the peptide 137 was replaced by A in peptide 250.

Examples of the variant peptide 219 replacing amino acids, 4-measure include, for example peptide 251, where K in position 2 of the peptide 219 replaced by A peptide 251.

Variant of the peptide with the substitution of amino acids, as described here, can be represented in the form of a selected peptide or in the form of chemically modified peptide, such as, for example, N-terminal amide, such as myristoylated, acetylene and the like, as described herein, such as, for example, C-terminal amide, such as an amide formed with ammonia, such as being both N-terminal and C-terminal amide.

When deletions are included in the amino acid sequence of the peptides of the invention in relation to a reference amino acid sequence, preferably at least 80% sequence identity between the amino acid sequence of the peptide and the reference amino acid sequence. Peptides having from 5 to 23 amino acids and consisting of one amino acid deletions relative to the reference peptide, will have 80% to about 96% (i.e. ~95,7%) sequence identity to the reference amino acid sequence. Peptides having from 10 to 23 amino acids and consisting of one amino acid deletions relative to the reference peptide, the Udut be from about 90% to about 96% (i.e. ~95,7%) sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and consisting of one amino acid deletions relative to the reference peptide, will be from about 95% to about 96% (i.e. ~95,7%) sequence identity to the reference amino acid sequence. Peptides having from 10 to 23 amino acids and comprising two amino acid deletions relative to the reference peptide, will be from about 80% to about 92% (i.e. ~91.3 percent) sequence identity to the reference amino acid sequence. Peptides having from 16 to 23 amino acids and comprising two amino acid deletions relative to the reference peptide, will be from about 87,5% to about 92% (i.e. ~91.3 percent) sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and comprising two amino acid deletions relative to the reference peptide, will be from about 90% to about 92% (i.e. ~91.3 percent) sequence identity to the reference amino acid sequence. Peptides having from 15 to 23 amino acids and comprising three amino acid deletions relative to the reference peptide, will be from about 80% to about 87% sequence identity to the reference amino acid sequentially the flesh. Peptides having from 20 to 23 amino acids and comprising three amino acid deletions relative to the reference peptide, will be from about 85% to about 87% sequence identity to the reference amino acid sequence. Peptides having from 20 to 23 amino acids and comprising four amino acid deletions relative to the reference peptide, will be from about 80% to about 83% (i.e. ~82,6%) sequence identity to the reference amino acid sequence.

As stated above, one or more amino acids of the peptides can also be chemically modified. Any known in the field of amino acid modification can be made with amino acids, peptides, using any known in the field of fashion.

In some embodiments the N-terminal and/or C-terminal amino acid may be modified. For example, the N-terminal amino acid peptides can be alkylated, liderovna or allerban N-terminal amino group (H2N-), and, for example, C-terminal amino acid peptides can be liderovna or etherification on the C-terminal carboxyl group (-COOH). For example, the N-terminal amino group can be modified by acylation by including any acyl or fatty acid acyl groups, amide forming, including acetyl g is the SCP (i.e. CH3-C(=O)-) or meritorious group. In some embodiments the N-terminal amino group may be modified by the inclusion of any acyl group having the formula-C(O)R, in which R represents a linear or branched alkyl group containing 1-15 carbon atoms, or may be modified by the inclusion of any acyl group having the formula-C(O)R1in which R1represents a linear alkyl group containing 1-15 carbon atoms. C-terminal amino acid of the peptides may also be chemically modified. For example, the C-terminal carboxyl group of the C-terminal amino acids can be chemically modified by the incorporation of an amino group instead of the hydroxyl group (i.e liderovna). In some embodiments the N-terminal and/or C-terminal amino acids not chemically modified.

The peptide can be allerban the amino group of the N-terminal amino acids, forming a N-terminal amide with an acid selected from the group consisting of:

(i)2-C24aliphatic carboxylic acids (saturated or not necessarily saturated) (for example, N-terminal amide of acetic acid, propanoic acid, butane acid, hexanoic acid, octanoic acid, decanoas acid, dodecanol acid, tetradecanoic acid (myristic acid), hexadecanoic acid, 9-hexadec the new acid, octadecanoic acid, 9-octadecenoic acid, 11-octadecenoic acid, 9,12-octadecadienoic acid, 9,12,15-octadecatrienoic acid, 6,9,12 - octadecatrienoic acid, eicosanol acid, 9-Aksenovo acid, 5,8,11,14-eicosatetraenoic acid, 5,8,11,14,17-eicosapentaenoic acid, docosenoic acid, 13-docosenoic acid, 4,7,10,13,16,19-docosahexaenoic acid, tetracosanoic acid and the like);

(ii) triperoxonane acid; and

(iii) benzoic acid; and

(iv)1-C24aliphatic alkylsulfonic acid, which forms an aliphatic alkylsulfonate, in which the structure of the carbon chain With1-C24aliphatic alkyl sulfonic acid is similar to the structure of the aliphatic alkyl chain carboxylic acid in the aliphatic alkyl carboxylic acids described above. For example, the peptide can be allerban using a carboxylic acid group, represented as (C1-C23)-alkyl-C(O)OH, by condensation with dehydration by activating the carboxyl group of the acid with the formation of amide represented as (C1-C23)-alkyl-C(O)-NH-peptide. Similarly, the sulfonamide can be formed by reaction of the derivatives of sulfonic acid (represented as a (C1-C23)-alkyl-S(O2)-X, where X is a halogen or OCH3or another joint which may leaving group) with N-terminal amino group with the formation of sulfonamida, represented as (C1-C23)-alkyl-S(O2)-NH-peptide.

In another embodiment of the N-terminal amino group of the N-terminal amino acids can be alkylated With1-C24aliphatic alkyl group, and the structure of the aliphatic alkyl groups such as described above. The alkylation may be effected, for example, aliphatic alkylhalogenide or aliphatic ether alkylsulfonic acid (mesilate, toilet etc), preferably using primary alkylhalogenide or primary ether alkylsulfonic acid. N-terminal amino acid may be modified by the terminal amino group by including any acyl or aliphatic acyl fatty acid group with amide formation, including acetyl group (i.e.- C(O)CH3), meritorious group, butanoyloxy group, hexanoyl, octanoyl group, technology group, dodecanoyl group, tetradecanoyl group, hexadecanoyl group, 9-hexadecanoyl group, octadecanoyl group, 9-octadecanoyloxy group, 11-octadecanoyloxy group, 9,12-octadecadienyl group, 9,12,15-octadecatrienoic group, 6,9,12-octadecatrienoic group, eicosanol group, 9-eicosanol group, 5,8,11,14-eicosatetraenoic group, 5,8,11,14,17-eicosapentaenoic the group, docosanol group, 13 - docosenoic group, 4,7,10,13,16,19-docosahexaenoyl group, tetracosanoic group, and each group is covalently attached to the terminal amino group of the peptide amide bond.

C-terminal carboxyl group of the C-terminal amino acids of the peptides of the invention can also be chemically modified. For example, the C-terminal amino acid may be chemically modified by reaction of the C-terminal carboxyl group of the peptide with the amine with the formation of amide groups, such as amide ammonium; amide With1-C24aliphatic alkylamine, preferably linear aliphatic alkylamine; amide hydroxyl-substituted C2-C24aliphatic alkylamine; linear amide 2-(C1-C24aliphatic alkyl)acetylamino; and amide omega-methoxy-poly(ethyleneoxy)n-etilenovomu (also designated as omega-methoxy-PEG-alpha-amino group or an omega-methoxy-(polyethylene glycol)-amino group), where n is from 0 to 10. C-terminal carboxyl group of the C-terminal amino acids of the peptide may also be in the form of an ether selected from the group consisting of ether With1-C24aliphatic Olkiluoto alcohol and ether 2-(omega-methoxy-poly(ethyleneoxy)nethanol-groups, where n is from 0 to 10.

C-terminal carboxyl group is PE is Chida, which can be represented by a formula of the peptide-C(O)OH, can also be liderovna by transformation into an activated form such as a carboxylic acid halide, carboxylic acid anhydride, N-hydroxysuccinimide ester, pentafluorophenyl ester (OPfp), 3-hydroxy-2,3-dihydro-4-oxo-benzo-trisonomy ether (ODhbt), and the like, to facilitate the reaction with ammonia or primary or secondary amine, preferably ammonia and primary amines, while it is preferable that other reactive group of the peptide were protected synthetic chemically compatible protective groups that are well known in the technical field of peptide synthesis, especially solid-phase peptide synthesis, such protective groups as benzyl ether, tert-butyl methyl ether, phenyl ether, etc. of the Obtained amide peptide can be represented by a formula of the peptide-C(O)-NR3R4(and amide is located on the C-end of the peptide), in which R3and R4independently selected from the group consisting of hydrogen; C1-C24of alkyl, such as methyl, ethyl, butyl, isobutyl, cyclopropylmethyl, hexyl, dodecyl, tetradecyl and the like, as described above.

C-terminal carboxylic acid C-terminal amino acids can also be converted into amide hydroxy-substituted C2-C24aliphatic alkylamine (where the hydroxyl is Naya group attached to the carbon atom lighter than the nitrogen atom of the amino group)such as 2-hydroxyethylamine, 4-hydroxyethylamine and 12-hydroxycobalamin and the like.

C-terminal carboxylic acid can also be converted into amide hydroxyl substituted C2-C24aliphatic alkylamine, in which the hydroxyl group can be azetilirovanna, forming an ester with2-With24aliphatic carboxylic acid as described above. Preferably amide peptide on the C-end of the peptide represented by the formula peptide-C(O)-R5R6where R5is hydrogen, R6selected from the group consisting of hydrogen, C1-C24aliphatic alkyl and hydroxy-substituted C2-C24the alkyl.

C-terminal carboxyl group of the C-terminal amino acids can also be converted into a linear amide 2-(C1-C24aliphatic alkyl)oxyethylene. This amide can be obtained, for example, by reaction With1-C24aliphatic alcohol with potassium hydride in diglyme with 2 chloroethanol to get linear With1-C24aliphatic alkylation, which can be converted into amine by oxidation to the aldehyde followed by reductive amination to an amine (for example, using ammonia), or by transformation in alkylhalogenide (for example, using thionyl chloride) followed by treatment of the amino is, such as ammonia.

C-terminal carboxylic acid C-terminal amino acids can also be converted into amide omega-methoxy-poly(ethyleneoxy)n-ethylamine, where n is from 0 to 10, which can be obtained from the corresponding omega-methoxy-poly(ethyleneoxy)nethanol, for example, by conversion of the alcohol to the amine as described above.

In another embodiment of the C-terminal carboxyl group can be converted into amide peptide represented by the formula peptide-C(O)-NR7R8in which R7is hydrogen, R8represents a linear 2-(C1-C24aliphatic alkyl)oxyethyl group, in which1-C24aliphatic alkyl portion is the same as described above, and includes such groups as methoxyethyl (i.e. CH3O-CH2CH2-), 2-dodecyloxy and the like; or R7is hydrogen, R8is omega-methoxy-poly(ethyleneoxy)n-ethyl group, where n poly(ethylenoxy) part is from 0 to 10, that is formed groups such as 2-methoxyethyl (i.e. CH3O-CH2CH2-), omega-methoxyethoxymethyl (i.e. CH3O-CH2CH2O-CH2CH2-) until CH3O-(CH2CH2O)10-CH2CH2-.

C-terminal carboxyl group of the C-terminal amino acids of the peptide may also be in the form of ether1-the 24aliphatic Olkiluoto alcohol, and aliphatic alkyl portion of the alcohol such as described above. C-terminal carboxyl group of the C-terminal amino acids of the peptide may be in the form of ether 2-(omega-methoxy-poly(ethyleneoxy)n)-ethanol group, where n is from 0 to 10, which can be obtained from the reaction of 2-methoxyethanol in the form of 2-methoxyethanol sodium with the stoichiometric quantity of ethylene oxide and stoichiometric quantity depends on the size n.

Side chain amino acid peptides can also be chemically modified. For example, the phenyl group of phenylalanine or tyrosine may be substituted by the Deputy selected from the group consisting of:

With1-C24aliphatic alkyl groups (i.e. linear or branched and/or saturated or unsaturated, and/or containing cyclic group, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, cyclopropyl, 2-methylcyclopropyl, cyclohexyl, octyl, decyl, dodecyl, hexadecyl, octadecyl, eicosanol, docosanol, tetracosane, 9-hexadecanoyl, 9-octadecenoyl, 11-octadecenoyl, 9,12-octadecadienyl, 9,12,15-octadecatrienoic, 6,9,12-octadecatrienoic, 9-eicosene, 5,8,11,14-eicosatetraenoic, 5,8,11,14,17-eicosapentaenic, 13-docosanol and 4,7,10,13,16,19-docosahexaenic;

With1-C24aliphatic alkyl groups, semese the Noah hydroxyl group at least one carbon atom away from the site of unsaturation, examples of such hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxydiphenyl and the like;

With1-C24aliphatic group, a substituted hydroxyl group, which etherification2-C24aliphatic carboxyl group of such acids as acetic acid, butane acid, hexanoic acid, octanoic acid, cekanova acid, dodekanisa acid, tetradecanoic acid, hexadecanol acid, 9-hexadecanoate acid, octadecanoic acid, 9-octadecenoate acid, 11-octadecenoyl acid, 9,12-octadecadienoic acid, 9,12,15-octadecatrienoic acid, 6,9,12-octadecatrienoic acid, Aksenova acid, 9-Aksenova acid, 5,8,11,14-eicosatetraenoic acid, 5,8,11,14,17-eicosapentaenoic acid, docosanoate acid, 13-docosanoate acid, 4,7,10,13,16,19-docosahexaenoic acid, tetracosanoic acid and the like, dicarboxylic acid such as succinic acid, or hydroxycitrate, such as lactic acid, where the total number of carbon atoms of the ether substituents is in the range from 3 to 25;

halogen, such as fluorine-, chlorine - and iodine; nitro; amino, such as NH2methylamino, dimethylamino; trifluoromethyl; carboxyl (-COOH);

With1-C24alkoxy (such as may be formed by alkylation of tyrosine), such as methoxy, this is XI, propyloxy, isopropoxy, bucalossi, isobutoxy, cyclopropylamino, 2-methylcyclopropyl, cyclohexyloxy, octyloxy, decyloxy, dodecyloxy, hexadecylamine, octadecylamine, eicosanoic, docosanoic, tetracosanoic, 9 hexadecenoic, 9 octadecanoyloxy, 11 octadecanoyloxy, 9,12-octadecadienoate, 9,12,15-octadecatrienoic, 6,9,12-octadecatrienoic, 9 eicosenoic, 5,8,11,14-eicosatetraenoate, 5,8,11,14,17-eicosapentaenoate, 13 docosenoic and 4,7,10,13,16,19-docosahexaenoate; and

With2-C24hydroxyalkyloxy, such as 2-hydroxyethyloxy, and their esters with carboxylic acids, as described above, or with triperoxonane acid.

The hydroxyl group of serine can be etherification with substituent selected from the group consisting of:

group C2-C24aliphatic carboxylic acids, such as described above;

group triperoxonane acids;

the group of benzoic acid.

The Epsilon amino group of lysine can be chemically modified, for example, through the formation of amide with: group C2-C24aliphatic carboxylic acids (for example, by reaction of the amine with a chemically activated form of a carboxylic acid such as the acid chloride acid anhydride, N-hydroxysuccinimide ester, pentafluorophenyl ester (OPfp), 3-hydroxy-2,3-dihydro-4-oxo-benzo-trisonomy (ODhbt) EF the R, and the like), as described above, or a group of benzoic acid or group of amino acids. In addition, the Epsilon amino group of lysine can be chemically modified by alkylation of one or two1-C24aliphatic alkyl groups.

Carboxyl group of glutamic acid may be modified by formation of amide with an amine, such as ammonia, With1-C24primary aliphatic alkylamine (the alkyl part of which is such as described above), including methylamine; or the amino group of amino acids.

Carboxyl group of glutamic acid may be modified by formation of ether with1-C24aliphatic hydroxyl group, as described above, preferably the ether is formed with a primary alcohol With1-C24aliphatic alkyl, such as methanol, ethanol, propan-1-ol, n-dodecanol and the like, as described above.

The peptides of the invention have the mucin-inhibiting effect and/or mucus-inhibitory effect in the introduction of a mammal in a mucin - and/or mucus-inhibiting amount. The peptides may also be (1) greater mucin-inhibiting effect in the mammal than MANS peptide, with the introduction in equal concentrations, (2) greater mucus-inhibitory effect in a mammal, than has the MANS peptide, with the introduction in equal concentrations, and/and and (3) to have a greater solubility in water, than MANS peptide.

And MARCKS peptide and MANS peptide each includes meritorious group associated with the amino group of the N-terminal amino acid by an amide bond. However, as revealed here, inhibiting the hypersecretion of mucin activity of the peptides of the invention are not related to the presence of myristoleic group at the N-terminal amino acid sequence of the peptide. Actually, I found that certain peptides of the invention that do not contain N-terminal chemical modification, are inhibitory hypersecretion of mucin activity. Also found that certain peptides of the invention, which contain the N-terminal chemical modification with groups other than myristoleic, such as acetyl group at the N-terminal amide, show inhibitory hypersecretion of mucin activity. Actually, N-terminal acetylated version of the MANS peptide (not monitorowania) can manifest in the methods of the invention inhibitory hypersecretion of mucin activity. In addition, the peptide amino acid sequence comprising the amino acid sequence of the MANS peptide and its variants, as described here, can manifest in the methods of the invention inhibitory hypersecretion of mucin activity.

In one aspect this invention provides a method of inhibiting mucin hypersecretion in a mammal, comprising the introduction is of the mammalian peptide, inhibits mucin hypersecretion in an amount which inhibits the hypersecretion of mucin, and the peptide has an amino acid sequence selected from the group consisting of: (a) amino acid sequences that have sequence GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO. 1); and (b) amino acid sequence essentially identical to the sequence defined in (a); in which N-terminal amino acid of the peptide is not monitorowania, and C-terminal amino acid of the peptide is optionally independently chemically modified, and the peptide has the effect of inhibiting mucin hypersecretion when administered to a mammal in number, the inhibitory hypersecretion of mucin. In one embodiment of the N-terminal amino acid of the peptide is preferably azetilirovanna. In another embodiment, the peptide exhibits at least one of the properties: (a) greater effect of inhibiting mucin hypersecretion in a mammal, than shows SEQ ID NO. 1, in which N-terminal amino acid monitorowania, with the introduction of the specified mammal in equal concentrations, or (b) greater solubility in water than that of SEQ ID NO. 1, in which N-terminal amino acid monitorowania, at equal concentrations in the same liquid.

In some embodiments the N-terminal amino group can be modified by incorporating acyl group having the th formula-C(O)R, in which R represents a linear or branched alkyl group having from 1 to 15 carbon atoms, or may be modified by incorporating acyl group having the formula C(O)R1in which R1represents a linear alkyl group having from 1 to 15 carbon atoms. C-terminal amino acid of the peptides may also be chemically modified. For example, the C-terminal carboxyl group of the C-terminal amino acids can be chemically modified by the incorporation of an amino group in place of the hydroxyl group (i.e liderovna). In some embodiments the N-terminal and/or C-terminal amino acids not chemically modified.

In some embodiments the peptides can have a greater half-life in bronchoalveolar lavagno fluid (BALF)than in plasma, and may also have a greater half-life in the mucus of the person (for example, in the mucus of a person with cystic fibrosis)than in plasma.

The peptides can be used in methods of inhibiting the secretion of mucin and/or production of mucus in mammals, the treatment of hypersecretion of mucin and/or mucus in mammals, the treatment of pulmonary diseases of mammals, causing hypersecretion of mucin and/or mucus (such as, for example, asthma, chronic bronchitis, COPD and cystic fibrosis). The conditions that may be approaching them to ensure peptides, include, but are not limited to, inflammatory, viral or bacterial respiratory disease (e.g. asthma, chronic obstructive pulmonary disease (COPD), colds, rhinitis, acute or chronic bronchitis, pneumonia and titanicby cough), allergic conditions (e.g., atopy, allergic inflammation), bronchiectasis, emphysema, bronchial asthma and certain genetic condition (e.g., cystic fibrosis). The peptides can also be suitable for treating conditions and diseases, and for use in the methods described in patent application U.S. No. 10/180753 (publication number US 2003/0013652) and 09/256154 and international application PCT/US00/05050 (international publication number WO 00/50062), the full content of which is given here by reference.

In addition to hypersecretion of mucin associated with the disease, the term hypersecretion of mucin also includes ATP-induced mucin hypersecretion as well as hypersecretion of mucin induced secretory tool, and induced hypersecretion of mucin.

In the preferred embodiment of the invention is a peptide which inhibits the hypersecretion of mucin and having the amino acid sequence of 4-23 consecutive amino acids of the reference amino acid sequence of PEPTIDE 1, peptide, ingebi the existing hypersecretion of mucin, chosen from:

(a) the group consisting of peptide 2, peptide 3, peptide 4, peptide 5 peptide 6, peptide 7 peptide 8, peptide 9, peptide 10, peptide 11, peptide 12, peptide 13, peptide 14, peptide 15, peptide 16, peptide 17, peptide 18, peptide 19, peptide 20, peptide 21, peptide 22, peptide 23, peptide 24, peptide 25, peptide 26, peptide 27, peptide 28, peptide 29, peptide 30, peptide 31, peptide 32, peptide 33, peptide 34, 35 peptide, peptide 36, 37 peptide, peptide 38, peptide 39, 40 peptide, peptide 41, 42 peptide, peptide 43, 44 peptide, peptide 45, peptide 46, peptide 47, peptide 48, peptide 49, peptide 50, peptide 51, peptide 52, peptide 53, peptide 54, peptide 56, 57 peptide, peptide 58, peptide 59, peptide 60, peptide 61, 62 peptide, peptide 63, peptide 64, peptide 67, peptide 68, peptide 69, peptide 70, peptide 71, peptide 72, peptide 73, peptide 74, peptide 75, peptide 79, peptide 80, peptide 81, peptide 82, peptide 83, peptide 84, peptide 85, peptide 86, peptide 87, peptide 92, peptide 93, peptide 94, peptide 95, peptide 96, peptide 97, peptide 98, 99 peptide, peptide 100, peptide 106, peptide 107, peptide 108, peptide 109, peptide 110, peptide 111, peptide 112, peptide 113, peptide 114, peptide 122, peptide 123, peptide 124, peptide 125, 126 peptide, peptide 127, peptide 128, peptide 129, peptide 139, 140 peptide, peptide 141, peptide 142, peptide 143, peptide 144, peptide 145, peptide 157, peptide 158, peptide 159, peptide 160, peptide 161, 162 peptide, peptide 176, peptide 177, PEP is IDA 178, peptide 179, peptide 180, peptide 196, peptide 197, peptide 198, peptide 199, 217 peptide and peptide 219, as described above; and

(b) amino acid sequence having 80%-96% sequence identity with the sequence defined in (a);

where the amino group of the N-terminal amino acids of amino acid sequence of the peptide that inhibits mucin hypersecretion, optionally covalently linked by an amide bond with a carboxylic acid selected from the group consisting of myristic acid and acetic acid, and

where the carboxyl group of the C-terminal amino acids of amino acid sequence of the peptide that inhibits mucin hypersecretion, optionally covalently linked by an amide bond with ammonia.

The peptide can be injected locally or systemically (for example, in the form of a pharmaceutical composition of the peptide of the invention and a pharmaceutically acceptable carrier and may be entered in any part of the body of a mammal, including, but not limited to, those parts of the body that produce mucus and/or mucin (for example, preferably in the respiratory passages, nasal cavity, oral cavity, trachea, lungs, gastrointestinal tract, eyes, reproductive tract, etc). The peptides can be introduced in various ways, including, but not limited to, local injection, parenteral administration, rectal the e introduction, pulmonary introduction, nasal introduction, inhalation, insufflation and oral administration. Pulmonary administration can be performed, for example, aerosol, spray, inhaler dry powder, metered dose inhaler and the like.

The peptides can be prepared and introduced as a pharmaceutical composition, suitable for any pharmaceutically effective route of administration. The peptides of the invention (or pharmaceutical composition based on them) can be in a form suitable for oral use (for example, in the form of tablets, pellets, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example, in the form of creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example, in the form of finely ground powder or a liquid aerosol), or for the introduction of insufflating (for example, in the form of finely crushed powder)or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous or intramuscular dosing or as suppository for rectal dosing).

The peptides can be introduced into the respiratory tract of the mammal by any suitable method, including generating Aero is Olya, which includes the peptide. The spray can be solid or liquid and may be water-based. Suitable particle size such aerosol can be produced in various ways known in the art. Particles of this aerosol is typically less than 50 microns in size, preferably in the range from about 0.1 micrometer to about 10 micrometers in size, more preferably in the range from about 1 micrometer to about 10 micrometers in size, and even more preferably in the range from about 2 micrometers to about 7 micrometers in size, and preferably from about 2 to about 7 micrometers.

Nasal introduction of the peptide or pharmaceutical composition of the peptide) can be performed, for example, by applying a spray of an aqueous solution or suspension of the peptide, or by applying aqueous solutions or suspensions of the peptide as drops or droplets, such as a dropper or pipette, for example, in the form of pharmaceutically acceptable, preferably sterile saline solution (NaCl), which may not necessarily be superiror. Sterilization of an aqueous solution may be performed, for example, by using sterile filtration of aqueous solution comprising the peptide, optionally in the presence of pharmaceutically acceptable additives such as preservatives,antioxidants, buffer salts, tone modifiers and the like. Sterilization of the water suspension comprising the peptide can be performed, for example, by irradiation or sterile filtration, when the suspension material particle size less than 0.2 micrometer, for example, the suspension comprising a micelle, a liposome, such as stabilized by phospholipids of the liposome, or a similar particle that can pass through a filter of 0.2 micrometer. An aqueous solution (e.g., isotonic saline or hypertonic saline, and optionally sugar, which is not chemically react with the peptide) a therapeutically effective amount of the peptide of the invention may be sterilized, for example, by using sterile filtration, and then an aliquot containing a therapeutically effective amount of a peptide is placed in a sterile vessel, lyophilized with the formation of a dry powder and sealed preferably in the presence of an inert atmosphere or gas, using a sterile cork or cap. Before applying the dry powder can be rehydration by adding sufficient quantity of water for the formation of a single dose of isotonic, which can be introduced as an aerosol into the respiratory tract of a mammal.

The amount of peptide or pharmaceutical composition based on it, the input melicope the surrounding, may vary depending on numerous factors, including, but not limited to, peptide, the type of mammal, the degree of disease of a mammal, the weight of the mammal and the age of the mammal. In addition, it may be desirable to maintain some level of secretion of mucin/production of mucus (e.g., normal or basal level) in a mammal, the past treatment of the peptide. In one embodiment man is preferred mammals from the point of view of the need for treatment, and the number of the input peptide that is entered in single or multiple dose sufficient to inhibit the hypersecretion of mucin. A single dose of the peptide of the present invention may vary from about 0.1 mg to 30 mg per kilogram of body weight, preferably from about 0.1 mg to about 15 mg per kilogram of body weight, and more preferably from about 0.1 mg to about 7.5 mg per kilogram of body weight. The number of doses and duration of treatment depends on the patient's response to treatment. Adjust the number of unit doses, number of doses and duration of treatment may be determined by a health care worker depending on the symptoms and the severity of these symptoms.

In one embodiment, the selected peptide consists of less than 24 sequentially location is the R amino acids and has the amino acid sequence, selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids (or in other embodiments 8 to 14 consecutive amino acids, or 10-12 consecutive amino acids) of the reference sequence PEPTIDE 1, where the amino acid sequence begins with the N-terminal amino acids of the reference sequence; (b) a sequence essentially similar to the amino acid sequence defined in (a); (c) amino acid sequence having from 4 to 23 consecutive amino acids (or in other embodiments 8 to 14 consecutive amino acids, or 10-12 consecutive amino acids the reference sequence PEPTIDE 1, where the amino acid sequence ends with the C-terminal amino acid of the reference sequence; and (d) sequence essentially similar to the amino acid sequence defined in (C). N-terminal amino group of the N-terminal amino acids and/or C-terminal carboxyl group of the C-terminal amino acids may not necessarily be chemically modified as follows:

(1) N-terminal amino group of the N-terminal amino acids of the peptide are not necessarily presented in the form of an amide selected from the group consisting of:

amid the 2-C24aliphatic carboxylic acid,

amide triperoxonane acid,

amide benzoic acid, and

amide With1-C24aliphatic alkylsulfonic acid; or

N-terminal amino group of the N-terminal amino acids of the peptide optionally alkylated by a group selected from the group consisting of:

With1-C24aliphatic alkyl groups,

linear 2-(C1-C24aliphatic alkyl)oxyethyl group,

omega-methoxy-poly(ethyleneoxy)n-ethyl group, where n is from 0 to 10; and

(2) the C-terminal carboxyl group of the C-terminal amino acids of the peptide are not necessarily presented in the form of an amide selected from the group consisting of:

amide ammonium,

amide With1-C24aliphatic alkylamine,

amide hydroxy-substituted C2-C24aliphatic alkylamine,

amide linear 2-(C1-C24aliphatic alkyl)acetylamino, and

amide omega-methoxy-poly(ethyleneoxy)n-etilenovomu, where n is from 0 to 10; or

C-terminal carboxyl group of the C-terminal amino acids of the peptide are not necessarily presented in the form of an ether selected from the group consisting of:

ether1-C24aliphatic Olkiluoto alcohol,

ether 2-(omega-methoxy-poly(ethyleneoxy)n)-ethanol group,where n is from 0 to 10.

The peptide has the effect of inhibiting the release of mucin when administered to a mammal in a mucin-inhibiting amount. The peptide may also have greater mucin-inhibiting effect in the mammal than the MANS peptide, with the introduction in equal concentrations and/or greater solubility in water than the MANS peptide.

In another embodiment, the selected peptide consists of less than 24 amino acids and has the amino acid sequence of the variant amino acid sequence having from 4 to 23 consecutive amino acids (or, in other embodiments, 8-14 consecutive amino acids, or 10-12 consecutive amino acids) of the reference sequence PEPTIDE 1, where the amino acid sequence begins with the N-terminal amino acids of the reference sequence, or where amino acid sequence is over the C-terminal amino acid of the reference sequence. N-terminal amino group of the N-terminal amino acids and/or C-terminal carboxyl group of the C-terminal amino acids may not necessarily be chemically modified as follows:

(1) N-terminal amino group of the N-terminal amino acids of the peptide are not necessarily presented in the form of an amide selected from the group consisting of:

amide With2-C24aliphatic carboxylic acid is you,

amide triperoxonane acid,

amide benzoic acid, and

amide With1-C24aliphatic alkylsulfonic acid; or

N-terminal amino group of the N-terminal amino acids of the peptide optionally alkylated by a group selected from the group consisting of:

With1-C24aliphatic alkyl groups,

linear 2-(C1-C24aliphatic alkyl)oxyethyl group,

omega-methoxy-poly(ethyleneoxy)n-ethyl group, where n is from 0 to 10; and

(2) the C-terminal carboxyl group of the C-terminal amino acids of the peptide are not necessarily presented in the form of an amide selected from the group consisting of:

amide ammonium,

amide With1-C24aliphatic alkylamine,

amide hydroxy-substituted C2-C24aliphatic alkylamine,

amide linear 2-(C1-C24aliphatic alkyl)acetylamino, and

amide omega-methoxy-poly(ethyleneoxy)n-etilenovomu, where n is from 0 to 10; or

C-terminal carboxyl group of the C-terminal amino acids of the peptide are not necessarily presented in the form of an ether selected from the group consisting of:

ether1-C24aliphatic Olkiluoto alcohol,

ether 2-(omega-methoxy-poly(ethyleneoxy)n)-ethanol group, where n is from 0 to 10.

The peptide has a mucin-in ibrowse effect when administered to a mammal in a mucin-inhibiting amount. The peptide may also have greater mucin-inhibiting effect in the mammal than the MANS peptide, with the introduction in equal concentrations and/or greater solubility in water than the MANS peptide.

In a further embodiment of the method of inhibiting mucin hypersecretion in a mammal includes an introduction to a mammal selected peptide in the mucin-inhibiting amount that inhibits the secretion of mucin. The selected peptide consists of less than 24 amino acids and has an amino acid sequence selected from the group consisting of: (a) amino acid sequence having from 4 to 23 consecutive amino acids (or, in other embodiments, 8-14 consecutive amino acids, or 10-12 consecutive amino acids) of the reference sequence PEPTIDE 1, where the amino acid sequence begins with the N-terminal amino acids of the reference sequence; (b) a sequence essentially similar to the amino acid sequence defined in (a); (c) amino acid sequence having from 4 to 23 consecutive amino acids (or, in other embodiments, 8-14 consecutive amino acids, or 10-12 consecutive amino acids) of the reference sequence PEPTIDE 1, where the amino acid sequence engaged in the W C-terminal amino acid of the reference sequence; and (d) sequence essentially similar to the amino acid sequence defined in (C). N-terminal amino group of the N-terminal amino acids and/or C-terminal carboxyl group of the C-terminal amino acids may not necessarily be chemically modified as follows:

(1) N-terminal amino group of the N-terminal amino acids of the peptide are not necessarily presented in the form of an amide selected from the group consisting of:

amide With2-C24aliphatic carboxylic acid,

amide triperoxonane acid,

amide benzoic acid, and

amide With1-C24aliphatic alkylsulfonic acid; or

N-terminal amino group of the N-terminal amino acids of the peptide optionally alkylated by a group selected from the group consisting of:

With1-C24aliphatic alkyl groups,

linear 2-(C1-C24aliphatic alkyl)oxyethyl group,

omega-methoxy-poly(ethyleneoxy)n-ethyl group, where n is from 0 to 10; and

(2) the C-terminal carboxyl group of the C-terminal amino acids of the peptide are not necessarily presented in the form of an amide selected from the group consisting of:

amide ammonium,

amide With1-C24aliphatic alkylamine,

amide hydroxy-substituted C2-C24aliphatic alkylamine,

amide linear 2-(C1/sub> -C24aliphatic alkyl)acetylamino, and

amide omega-methoxy-poly(ethyleneoxy)n-etilenovomu, where n is from 0 to 10; or

C-terminal carboxyl group of the C-terminal amino acids of the peptide are not necessarily presented in the form of an ether selected from the group consisting of:

ether1-C24aliphatic Olkiluoto alcohol,

ether 2-(omega-methoxy-poly(ethyleneoxy)n)-ethanol group, where n is from 0 to 10.

The peptide may have a greater mucin-inhibiting effect in the mammal than the MANS peptide, with the introduction in equal concentrations and/or greater solubility in water than the MANS peptide.

The peptides of the invention can be obtained by any suitable method, including the method of solid-phase peptide synthesis, such as, for example, using fluorenylmethoxycarbonyl (FMoc) strategy and a suitable peptide synthesizer, such as a peptide synthesizer CS-Bio, or using tert-butyloxycarbonyl (Boc) strategy and a suitable peptide synthesizer, such as the ABI 430A Peptide Synthesizer. Protected amino acids suitable for use in FMoc and Boc synthesis, such as those available from Calbiochem, EMD Bioscience, San Diego, CA. In solid-phase peptide synthesis of C-terminal carboxyl group of the target peptide covalently attached to the polymer is odlozte in the form of N-alpha-protected amino acids. The protective group of N-alpha-amino group is then removed and condense the second N-alpha-protected amino acid with a fixed amino acid through the formation of amide bond with an unprotected N-alpha-amino group of amino acids associated with the resin. These stages repeat with the appropriate protected amino acids in the target peptide sequence until the target sequence. At the end of the synthesis the relationship between the C-terminal amino acid and the polymer substrate is cleaved, releasing the peptide. The peptide can be isolated and purified HPLC. Used cleaning methods HPLC include ion-exchange chromatography and reversed-phase chromatography. Solutions of peptides can be evaporated or liofilizovane to get the selected peptide in solid form. Peptides containing oxidizable groups such as the remains of methionine, cysteine, tryptophan, preferably supported in an atmosphere free from oxygen, and when they are included in the composition, and stored in the form of a solution or suspension, the use of free oxygen from the solvent.

Condensation of activated esters with aminocom.com peptide bound to the resin during the synthesis, can be produced, for example, using an excess, e.g., 4-fold excess of amino acids and benzotriazol-1-yl-oxy-Tris-dimethylamino-phosphonium exaft is hosphate, and excess, such as 6-fold excess of N,N-diisopropylethylamine. Peptides during FMo synthesis otscheplaut from the resin using triperoxonane acid/thioanisole/triisopropylsilane/methanol (for example, in the ratio of 90:5:2,5:2,5, vol./about./about./about.) at 20°C for 4 hours, and peptides during Boc synthesis otscheplaut using, for example, HF/anisole (9:1, vol/about.) at 4°C for 1 hour.

In FMoc peptide strategy for the first FMoc amino acid is fixed on an insoluble polymer substrate through cyclotouring linker. Removing protection perform the processing of amino acid base, such as piperidine. The second FMoc amino acid condense using pre-activated compounds or in situ activation (the condensation reaction can be performed in situ using activating reagents, such as known in peptide chemistry, DCC, HBTU, TBTU, BOP, BOP-Cl, and the like). After the target peptide was synthesized with a peptide bound to the resin, remove the protection, and it is cleaved from the solid substrate by acid hydrolysis of weak acids such as triperoxonane acid (TFA) or TMSBr in the presence of an absorber, such as, for example, tilne connection, phenol and water. In one aspect before removing the protection of functional groups in the side chains and cleavage from the resin terminal amino group of the peptide can be processed with the carboxylic acids (for example, aliphatic carboxylic, triperoxonane, benzoic and the like), for example, varieties of aliphatic carboxylic acids, such as an activated form of aliphatic carboxylic acids, such as pendaftar ether, similar to the method of formation of the peptide bond with the formation of amide carboxylic acid, or varieties of aliphatic sulfonic acids (such as sulfonyl chloride), with the formation of sulfonamida on the N-end of the peptide. Alternatively, the N-terminal amine can be alkylated, for example, aliphatic alkylating agent (for example, aliphatic mesilate or tosylate)obtained by reaction of the corresponding sulphonylchloride and bases, such as pyridine, aliphatic alcohol, and the alcohol can be obtained by reduction (for example, lithium aluminum hydride) aliphatic carboxylic acid. In another aspect, the corresponding D-amino acids (for example, up to four D-amino acids) with other L-amino acids can be used in the peptide synthesis procedure to obtain the peptide of the invention, which optionally may be chemically modified as described above. In another aspect, formed when the target peptide amino acid sequence, for example, solid-phase synthesis and when the selected group protecting the side chain, so that in order to keep the stage of cleavage of the ether and release of the peptide from the resin, the resulting protected peptide side chain and having a free carboxylic acid at the C-end of the peptide. C-terminal carboxylic acid can be converted into an activated ester (e.g., pentafluorophenyl ether and treated with the amine, such as ammonia, forming amide (presented as peptide-C(O)-NH2), or, as described above, processed aliphatic amine, forming aliphatic amide peptide, and any remaining protective groups can be removed to obtain the target peptide of the invention. In addition, the C-terminal carboxylic ester can be formed from the C-terminal carboxylic acid and aliphatic alcohol with the condensation with dehydration, such as using a carbodiimide reagent. Containing acidic functional groups of amino acids such as aspartic acid and glutamic acid can be converted into amides and esters in a manner analogous to the above-described method, and the Epsilon amino group of lysine can be transformed into amides and aliphatic amines, as described above for the chemistry of end amino groups.

Examples of protected amino acids using FMoc solid-phase synthesis of the peptides of the invention include the following non-limiting examples: pentafluorophenyl ester N-alpha-Fmoc-L-alanine; pentafluorophenyl ester N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxime the ZIL)-alanine; pentafluorophenyl ester N-alpha-Fmoc-glycine; pentafluorophenyl ester N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-glycine; g-2-phenylisopropyl ester N-alpha-Fmoc-L-glutamic acid; alpha-4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohex)-3-methylbutyl]amino}benzyl ester N-alpha-Fmoc-L-glutamic acid; gamma-4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohex)-3-methylbutyl]amino}benzyl ester N-alpha-Fmoc-L-glutamic acid; alpha-allyl ether of N-alpha-Fmoc-L-glutamic acid; g-benzyl ester N-alpha-Fmoc-L-glutamic acid; alpha-tert-butyl ester N-alpha-Fmoc-L-glutamic acid; gamma-tert-butyl ester N-alpha-Fmoc-L-glutamic acid; pentafluorophenyl ester of gamma-tert-butyl ester N-alpha-Fmoc-L-glutamic acid; panafcortelone ester N-alpha,Epsilon-Di-Fmoc-L-lysine; N-alpha,Epsilon-Di-Fmoc-L-lysine; N-alpha-1-(4,4-dimethyl-2,6-dioxocyclohex-1-ilidene)ethyl-N-Epsilon-Fmoc-L-lysine; N-alpha-Fmoc-L-lysine; N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-N-Epsilon-tert-butoxycarbonyl-L-lysine; N-alpha-Fmoc-N-Epsilon-1-(4,4-dimethyl-2,6-dioxocyclohex-1-ilidene)-3-methylbutyl-L-lysine; N-alpha-Fmoc-N-Epsilon-1-(4,4-dimethyl-2,6-dioxocyclohex-1-ilidene)-ethyl-L-lysine; N-alpha-Fmoc-N-Epsilon-2-chloro-CBZ-L-lysine; N-alpha-Fmoc-N-Epsilon-4-methyldecyl-L-lysine; N-alpha-Fmoc-N-Epsilon-acetyl-L-lysine; N-alpha-Fmoc-N-Epsilon-benzyloxycarbonyl-L-lysine; N-alpha-Fmoc-N-Epsilon-tert-Boc-L-l the zine; pentafluorophenyl ester N-alpha-Fmoc-N-Epsilon-tert-Boc-L-lysine; N-alpha-Fmoc-N-Epsilon-TRIFLUOROACETYL-L-lysine; N-alpha-Fmoc-4-chloro-L-phenylalanine; N-alpha-Fmoc-4-cyano-L-phenylalanine; N-alpha-Fmoc-4-fluoro-L-phenylalanine; N-alpha-Fmoc-4-nitro-L-phenylalanine; N-alpha-Fmoc-L-phenylalanine; pentafluorophenyl ester N-alpha-Fmoc-L-phenylalanine; N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-phenylalanine; N-alpha-Fmoc-N-alpha-methyl-L-phenylalanine; pentafluorophenyl ester N-alpha-Fmoc-L-Proline; N-alpha-Fmoc-gamma-(4,4'-dimethoxybenzyl)-L-glutamine; pentafluorophenyl ester N-alpha-Fmoc-gamma-trityl-L-glutamine; N-alpha-Fmoc-L-glutamine; pentafluorophenyl ester N-alpha-Fmoc-L-glutamine; N-alpha-Fmoc-N-gamma-trityl-L-glutamine; N-methoxy-N-methylamide N-alpha-Fmoc-NG-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-L-arginine; N-alpha-Fmoc-NG-2,2,4,6,7-pentamethylcyclopentadiene-5-sulfonyl-L-arginine; N-alpha-Fmoc-NG-2,2,5,7,8-pentamethylchroman-6-sulfonyl-L-arginine; N-alpha-Fmoc-NG-4-methoxy-2,3,6-trimethylbenzenesulfonyl-L-arginine; pentafluorophenyl ester N-alpha-Fmoc-NG-4-methoxy-2,3,6-trimethylbenzenesulfonyl-L-arginine; N-alpha-Fmoc-NG-nitro-L-arginine; N-alpha-Fmoc-NG-tosyl-L-arginine; N-alpha-Fmoc-O-(2-acetamido-2-deoxy-3,4,6-tri-O-acetyl-alpha-D-galactopyranosyl)-L-serine; N-alpha-Fmoc-L-serine; N-alpha-Fmoc-O-benzyl-L-phosphoserine; N-alpha-Fmoc-O-benzyl-L-serine; N-alpha-Fmoc-O-tert-butyl-L-serine; N-alpha-Fmoc-O-tert-is util-L-serine N-hydroxysuccinimide; N-alpha-Fmoc-O-trityl-L-serine; N-alpha-Fmoc-L-threonine; N-alpha-Fmoc-O-benzyl-L-posttraining; N-alpha-Fmoc-O-benzyl-L-threonine; N-alpha-Fmoc-O-tert-butyl-L-threonine; N-alpha-Fmoc-O-trityl-L-threonine; N-alpha-Fmoc-O-(2-acetamido-2-deoxy-3,4,6-tri-O-acetyl-alpha-D-galactopyranosyl)-L-threonine; N-alpha-Fmoc-L-valine; pentafluorophenyl ester N-alpha-Fmoc-L-valine; N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-valine; pentafluorophenyl ester N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-valine; N-alpha-Fmoc-N-alpha-methyl-L-valine; N-alpha-Fmoc-O-(bis-dimethylamino-phosphono)-L-tyrosine; N-alpha-Fmoc-L-tyrosine; N-alpha-Fmoc-O-2,6-dichlorobenzyl-L-tyrosine; N-alpha-Fmoc-O-2-bromo-CBZ-L-tyrosine; N-alpha-Fmoc-O-2-chlorotrityl-L-tyrosine; N-alpha-Fmoc-O-benzyl-L-phosphotyrosine; N-alpha-Fmoc-O-methyl-L-tyrosine; N-alpha-Fmoc-O-phospho-L-tyrosine; N-alpha-Fmoc-O-tert-butyl-L-tyrosine; pentafluorophenyl ester N-alpha-Fmoc-O-tert-butyl-L-tyrosine; beta-1-adamantylamine ester N-alpha-Fmoc-L-aspartic acid; beta-2-adamantly ester N-alpha-Fmoc-L-aspartic acid; beta-2-phenylisopropylamine ester N-alpha-Fmoc-L-aspartic acid; beta-4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohex)-3-methylbutyl]-amino}benzyl ester N-alpha-Fmoc-L-aspartic acid; alpha-4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohex)-3-methylbutyl]-amino}benzyl ester N-alpha-Fmoc-L-aspartic acid; alpha-allyl ether of N-alpha-Fmoc-L-aspartic acid beta-benzyl e is Il N-alpha-Fmoc-L-aspartic acid; alpha-tert-butyl ester N-alpha-Fmoc-L-aspartic acid beta-tert-butyl ester N-alpha-Fmoc-L-aspartic acid; pentafluorophenyl ester of beta-tert-butyl ester N-alpha-Fmoc-L-aspartic acid; pentafluorophenyl ester of beta-1-adamantinoma ester N-alpha-Fmoc-L-aspartic acid; pentafluorophenyl ester of beta-2-adamantinoma ester N-alpha-Fmoc-L-aspartic acid; N-alpha-Fmoc-L-leucine; pentafluorophenyl ester N-alpha-Fmoc-L-leucine; N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-leucine; pentafluorophenyl ester N-alpha-Fmoc-N-alpha-(2-Fmoc-hydroxy-4-methoxybenzyl)-leucine; N-alpha-Fmoc-N-alpha-methyl-L-leucine; N-alpha-Fmoc-L-isoleucine; pentafluorophenyl ester N-alpha-Fmoc-L-isoleucine; N-alpha-Fmoc-N-alpha-methyl-L-isoleucine; N-alpha-Fmoc-beta-2,4,6-trimethoxybenzyl-L-asparagine; pentafluorophenyl ester N-alpha-Fmoc-beta-trityl-L-asparagine; N-alpha-Fmoc-L-asparagine; pentafluorophenyl ester N-alpha-Fmoc-L-asparagine; N-alpha-Fmoc-N-beta-(3,4,6-tri-O-acetyl-2-(acetylamino)-deoxy-2-beta-glyukopiranozil)-L-asparagine; N-alpha-Fmoc-N-beta-trityl-L-asparagine; N-alpha-Fmoc-N-im-methyldecyl-L-histidine; cyclohexylammonium salt of N-alpha-Fmoc-N-im-tert-Boc-L-histidine; N-alpha-Fmoc-N-im-tosyl-L-histidine; N-alpha-Fmoc-N-im-trityl-L-histidine; N-alpha-Fmoc-S-atsetamidometil-L-cysteine; pentafluorophenyl ester N-alpha-Fmoc-S-atsetamidometil-L-cysteine; N-alpha-Fmoc-S-benzyl-L-cysteine; N-alpha-Fmoc-S-p-m is kribensis-L-cysteine; N-alpha-Fmoc-S-p-methoxytrityl-L-cysteine; N-alpha-Fmoc-S-tert-butyl-L-cysteine; pentafluorophenyl ester N-alpha-Fmoc-S-tert-butyl-L-cysteine; N-alpha-Fmoc-S-tert-butylthio-L-cysteine; pentafluorophenyl ester N-alpha-Fmoc-S-tert-butylthio-L-cysteine; N-alpha-Fmoc-S-trityl-L-cysteine; pentafluorophenyl ester N-alpha-Fmoc-S-trityl-L-cysteine. These and other amino acid reagents, which are used in solid-phase synthesis of peptides, commercially available, for example, from Calbiochem Corporation. Aliphatic carboxylic acids are also available from Sigma-Aldrich Chemical Company.

The peptides can also be obtained by solid-phase synthesis using Boc strategy, in which the first Boc amino acid is attached to an insoluble polymeric substrate by HF-degradable linker. Removal of protection by removing the Boc group is accomplished by treatment of Boc amino acids TFA. Second Boc amino acid and then condensed, using pre-activated derivatives or in-situ activation. After the target peptide was synthesized with a peptide bound to the resin, remove the protection, and it unsnaps from the solid substrate by cleavage using strong acids such as HF, TFMSOTf or TMSOTf. Additive such compounds as thiol add to protect the peptide from the carbocations generated by splitting. These protective groups are compatible with the HF splitting: Ag(Mts); Cys(4-MeOBzl); His(Z); Arg(Tos); Glu(OBzl); Lyz(Cl-Z); Asp(OBzl); Glu(OcHex); Ser(Bzl); Asp(OcHex); His(Bom); Thr(Bzl); Cys(Acm); His(Dnp); Trp(CHO); Cys(4-MeBzl); His(Tos); Tyr(Br-Z); Asp(OtBu); His(Trt). These protective groups are compatible with TFMSOTf cleavage: Arg(Mts); His(Bom); Met(O); Asp(OBzl); His(Dnp); Ser(Bzl); Cys(Acm); His(Tos); Thr(Bzl); Cys(4-MeBzl); His(Z); Trp(CHO); Glu(OBzl); Lyz(Cl-Z); Tyr(Br-Z). These protective groups are compatible with TMSOTf cleavage: Arg(Mts); Glu(OcHex); Trp(CHO); Arg(Mbs); His(Bom); Trp(Mts); Asp(OBzl); Lyz(Cl-Z); Tyr(Br-Z); Asp(OcHex); Met(O); Tyr(Bzl); Cys(Acm); Ser(Bzl); Tyr(Cl-Bzl); His(Bom); Thr(Bzl). Condensation of amino acids with carboxylic acids amines for the formation of peptide amide bonds can be made in the Boc strategy using carbodiimides, such as dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), tert-butyl methyl and tert-butylether-carbodiimide; BOP; PyBroP; PyBOP; HBTU; TBTU and HATU, and each of these reagents to activate the necessary Foundation, and each operates through the formation of symmetrical anhydride. Alternatively can be used reagents mixed anhydrides of carboxylic acid and carbonic acid or carboxylic and phosphinic acids prepared using the reaction of isobutyl or isopropylcarbamate and substituted phosphine chlorides with N-alpha-protected amino acid or N-carboxyanhydride (NCA) or can be used Leuchs anhydride. As Fmoc strategy, N-terminal amino group and C-terminal carboxylic acid can be chemically mo is inficirovanyh according to the strategy described above for the synthesis.

EXAMPLES

The invention will be further explained by the following illustrative examples, and have in mind that these examples, the invention is not limited. These examples present the results of testing specific peptides, which are typical representatives of peptides identified in the application. The peptides listed with all modifications of N-terminal amino group of the N-terminal amino acids and/or C-terminal carboxyl group of the C-terminal amino acids from the left and right sides of the amino acid sequence, respectively, and labeled Peptide # tested peptides. Table IX contains the key to peptide sequences. Myr - and Ac - abbreviations myristoyl and acetyl, respectively, which are covalently linked by an amide bonds with the N-terminal amino acid in the corresponding amino acid sequence of the peptide; -NH2means amide ammonium, which is covalently linked to the C-end amino acid sequence of the peptide.

Example 1A - relative effectiveness of test peptides in a murine model of asthma

I. Protocols and methods

The experiments were designed to test whether or not the MANS peptide and other test related peptides inhibit hypersecretion in rat Airways in vivo. Ovalbumin-sensibi sirovina murine model of allergic inflammation and asthma, used in these studies, is the same as described by Singer et al. (2004),supra. As a negative control, control peptide containing the N-terminal meritorious group and the same amino acids that MANS peptide, but located in a random order (i.e. random N-terminal sequence, RNS, myristoyl-peptide 232), was tested in parallel with the alleged active peptides. BP2 mice aged 6-8 weeks were immunized twice a week, by entering 1 μg of ovalbumin intravenously. After a 14-day sensitization, animals were treated with sprayed-ovalbumin showed that after 72 hours causes prolonged hyperplasia cells Goblet. At the time, equal to 72 hours, delivered secretory tool metafolin (60 mm), using a spray system Buxco, providing high-quality aerosol within 90 seconds. 15 minutes before the introduction of secretory means nutritarian was injected 50 μl of the test peptide in 3 different concentrations (10 ám, 100 ám and 140 ám). RNS peptide was tested only at the highest dose level (50 ál 140 μm solution). MANS as well as RNS peptide, were easily dissolved in 120 mm sodium acetate solution, pH 7. Various controls used in the experiment are summarized in Table I below. Each experiment was performed using 6 mice at each point, and each set of experiments n is tarali 3 times. To test variations from line to line, the experiment was repeated with Balb/C mice by a similar Protocol. Both stimulated and non-stimulated secretion of mucin in mice treated with 120 mm sodium acetate solution (data not shown)were identical with the saline control. After the introduction of methacholine animals were killed, and for analysis of secreted mucin was taken bronchoalveolar lavage (BAL) from 6 animals in each group.

Table I shows the basic Protocol of the experiment.

No
Table I - main Protocol metacholine-induced hypersecretion of mucin in the presence of test peptide
The test group#
Mice
ProcessingPeptidesIntroduction secretory tools
Saline control6Free from endotoxin solution of 0.9% NaClNoYes
Saline control6Free from endotoxin solution of 0.9% NaClNo No
The test peptide6Free from endotoxin solution of 0.9% NaCl + 140 μm each test peptideEach test peptideYes
The test peptide6Free from endotoxin solution of 0.9% NaCl + 100 μm of each test peptideEach test peptideYes
The test peptide6Free from endotoxin solution of 0.9% NaCl + 100 μm of each test peptideEach test peptideNo
The test peptide6Free from endotoxin solution of 0.9% NaCl + 10 mm each test peptideEach test peptideYes
The test peptide6Free from endotoxin solution of 0.9% NaCl + 10 mm each test peptideEach test peptide
The negative control peptide (RNS)6Free from endotoxin solution of 0.9% NaCl + 140 μm peptide RNSRNSYes

II. Analysis of BAL fluid

Mice were killed with translocation of the cervical vertebrae with consequent rapid exposure of the trachea and the setting of the catheter through a small incision. Collected 0.5 ml of fluid from bronchoalveolar lavage (BAL) and then 3 x dilution in 1 ml PBS containing PMSF (5 mm), EDTA (5 mm) and DTT (5 mm). BAL fluid was divided into cellular and acellular fractions by short centrifugation, prior to any analysis undertaken. Cellfree supernatant analyzed for the presence of mucin by ELISA method using anti-satinowye antibodies, which shows that they react with murine mucin. More precisely, in this analysis, we used mouse anti-MUC5AC antibodies that recognize the hydrocarbon portion of the secreted mucin. The data obtained for each BAL sample were normalized with respect to the total amount of protein, as determined in the analysis according to Bradford. The content of mucin reflected as a difference between the values obtained with anti-musinovym antibodies, and the values obtained with the control not musinovym and what citelli. All ELISA data was statistically processed using univariate analysis of variance (ANOVA). Reviewed experimental data significantly different from control when p<0,05.

Tables II, III, IV, V and VI summarize the effect of different peptides on the secretion of mucin in Balb/C and BP2 mice. As shown in table II, at 100 μm concentration of the test peptides secretion of mucin is in the range from 8% to 56% of control (i.e. in the absence of peptide).

Table II - Secretion of mucin in a murine model of asthma (Experiments on Balb/C mice)
The peptide (peptide # ID)% of controlThe average error
No peptide1004,2
MANS, myr-peptide 1to 33.85,8
RNS, myr-peptide 232965,2
myr-peptide 795010,8
myr-peptide 2332114,2
myr-peptide 23484,2
myr-peptide 235133,3
peptide 79103,3
myr-peptide 234-NH227,81,6
myr-peptide 10633,65,8
peptide 106to 33.83,3
myr-peptide 106-NH229,34,2
myr-peptide 236-NH2512,0
AC-peptide 106a 21.518,3
myr-peptide 137554,2
myr-peptide 137-NH2563,3
All peptides were tested at a concentration of 100 μm. All values are accurate relative to control (p<0,001), with the exception of myr-peptide 137 (p<0,1) and RS.

Table III - Secretion of mucin in a murine model of asthma (Experiments on Balb/C mice)
The peptide (peptide # ID)% of controlThe average error
No peptide1008,1
MANS, myr-peptide 14011,8
RNS, myr-peptide 23281**2,8
myr-peptide 11545,1
myr-peptide 3735**11,8
myr-peptide 794810,3
myr-peptide 1538*10,6
myr-peptide 4535**10,3
myr-peptide 916814,7
myr-peptide 153 507,3
All peptides were tested at a concentration of 100 ám.
*=the value of reliable relative to control (p<0,05); **=reliable values relative to control (p<0,01).

Table IV - Secretion of mucin in a murine model of asthma (Experiments on Balb/C mice)
The peptide (peptide # ID)% of controlThe average error
No peptide1005,3
MANS, myr-peptide 1 (100 μm)29**4,2
RNS, myr-peptide 232 (100 μm)1088,5
peptide 237 (10 μm)67*+8,5
peptide 237 (100 μm)8**the 3.8
myr-peptide 106 (10 μm)74*+10,6
myr-peptide 106 (100 μm)24** 9,6
peptide 106 (10 μm)64*4,2
peptide 106 (100 μm)40**9,6
myr-peptide 106-NH2(10 μm)67*+4,2
myr-peptide 106-NH2(100 µm)17**5,3
*=the value of reliable relative to control (p<0,05); **=reliable values relative to control (p<0.01), and +=value reliable relative to 100 μm treatment (p<0,05).

Table V - Secretion of mucin in a murine model of asthma (Experiments on BP2 mice) (% of control)
Secretion of mucin (% of control)
Peptide (Conc.)10 µm100 mm140 mcm
MANS peptide35128
RNS peptide- 100100

The experiments were designed to determine the duration of the test peptides in a murine model of asthma. As described in Method 1A, Balb/C mice were immunized with ovalbumin. After 14 days, animals were injected with ovalbumin in aerosol form. After 72 hours delivered by aerosolization secretory tool metafolin. The tested peptides (50 l 100 M solution) was injected nutritarian for 30 min, 60 min or 120 min before the introduction of methacholine. Animals were killed and took BAL analysis of secreted mucin. The results of this experiment are given in Table VI.

td align="center"> 100
Table VI - Duration of test peptides in mice (experiments on Balb/C mice) (% of control)
Peptide30 min60 min120 min
Control100100100
MANS, myr-peptide 174450
RNS, myr-peptide 232100100
Ac-peptide 106217270
peptide 106234857

Alternative test mouse model and a way of quantifying the mucin in the lungs of mice that are suitable to assess the activity of the peptides of the present invention. This method is described in Evans et al. (Am. J. Respir. Cell Mol. Biol. Vol.31, pp 382-394, 2004). Briefly, Balb/C female mice (age 5-8 weeks, 20-25 g each) sensibiliser weekly for four weeks by intraperitoneal injection of 100 μl of a solution containing 2.2 mg of aluminium alum and 20 μg of ovalbumin in normal saline. 7 days after the last intraperitoneal injection to mice injected through the introduction of aerosol over 30 minutes in 2.5% solution of ovalbumin dissolved in normal saline. The aerosol generating a spray of compressed air AeroMist CA-209 (CIS-US, Inc., Bedford, MA).

Three days after injection of ovalbumin in 50 μl of the test peptide is delivered for about 2-3 minutes in each nostril mouse aliquot of 10 µl. Fifteen minutes later, mice treated with about 5 minutes sprayed with 100 mm ATP solution. After 20 minutes of anaesthetize mice intraperitoneal inj what Ktsia mixture of ketamine, of xylazine and acepromazine, and light perfusion saline through the right heart ventricle to clear the blood from the lung tissue. Under deep anesthesia, the animals tracheostomies using 20-caliber cannula with a blunt tip, and killed by exsanguination through the abdominal aorta. The fixative (4% paraformaldehyde in 0.1 M phosphate buffer, pH 7.0) pour nutritarian under pressure of 10-15 cm, and light fixin situwithin 30 minutes, removed from the thoracic cavity and fixed overnight at 4°C. Light is placed in paraffin and cut into 6 μm sections.

For fluorescent labeling of tissue mucin stain, using the fluorescent Schiff staining with iodine acid (PAFS). First fabric oxidized in 1% iodine acid (10 min), washed, treated acriflavine fluorescent Schiff's reagent (0.5% acriflavin HCl weight/vol., 1% sodium metabisulfite weight/vol., of 0.01 N. HCl) for 20 min, washed twice in deionized H2Oh, and washed 2x 5 min in acid alcohol (0.1 G. of HCl in 70% ethanol). Slides are dehydrated in gradient ethanol and allow to air dry in the dark. Once dried PAFS-stained preparations paste to cover the glass preparative environment with Canadian balsam (50% resin of Canada balsam, 50% methyl salicylate; Fisher Cheicals).

For quantification of mucin PAFS-stained preparations examined under 40x lens. Fix image 10 of the axial regions of the bronchi, the camera control using MagnaFire 2.1 (Optronics). Play PAFS images taken by excitation of the samples using a two-step excitation filter (peaks at 500 nm and 573 nm), and the observation of samples carried out using two-stage emission filter with peaks at 531 nm (green) and 628 nm (red). For each area of the image is first obtained using only red absorbing channel camera (exposure 590 MS). The same image is then fixed using both channels of the camera - red and green (590 MS red, 450 MS green). For morphometric analysis, then measure the bulk density and the intensity of fluorescence staining of mucin using ImagePro Plus. Bulk density of staining of mucin in the epithelium of the respiratory tract stereological count. Briefly, the ratio of the painted surface area to total surface area of the epithelium, divided by the length of the border dimension, which is the product of the total surface area of the epithelium, the length of the basal membrane and the geometric constants of 4/π. As a result, data are presented as the amount of intracellular mucin content per unit area of surface is the base membrane. Secretion of mucin expressed as a fraction of total content of epithelium.

Example 1B - the impact of the introduction of the powdered peptide AC-peptide 106 on the mucin hypersecretion in a mouse model metaplasia cells Goblet and airway obstruction

The efficiency of the powdered peptide Ac-peptide 106 on the mucin hypersecretion in a mouse model of asthma was determined at two concentrations. Briefly, 5-8-week-old Balb/C mice were immunized with ovalbumin on a weekly basis, as described above in Example 1A, for 3 weeks. On the 28th day the mice were injected within 30 minutes sprayed with 2.5% ovalbumin in normal saline. Three days after injection of ovalbumin each group of mice (n=2) were injected powdered isotonic Ac-peptide 106 at concentrations of 10 mm or 30 mm for 1 hour using AeroMist nebulizer CA-209. Total average aerodynamic diameter of the particles made 1.49 μm (in the range 0.4-4.7 µm). For these concentrations of Ac-peptide 106 and for a given particle size calculated deposition Ac-peptide 106 in the lungs was 0.38 mg/kg body weight and 1.1 mg/kg body weight, respectively.

Introduction Ac-peptide 106 followed immediately by the introduction of aerosol secretory means adenosine triphosphate (ATP) (100 mm in saline) for 5 minutes. Mice were killed, lungs were collected for 20 minutes. The lungs were washed in saline solution, the anchor is ovali paraformaldehyde, placed in paraffin and sectionlevel. Conducted quantitative analysis sections for the presence of mucin according to a quantitative immunohistochemical method of Evans et al. (Am. J. Respir. Cell Mol. Biol. Vol.31, pp 382-394, 2004). The results showed that Ac-peptide 106 was effective as an inhibitor on 30-67% of ATP-induced mucin hypersecretion at concentrations of 0.38 and 1.1 mg/kg body weight, respectively.

Example 2 - Qualitative assessment of the solubility of the tested peptides in 0.5 normal saline

Various test peptides (1-5 mg each) were carefully weighed into individual glass 2 ml vials with screw caps and thereto was added an aliquot of 25 μl of 0.5 N. salt solution at 25°C and normal pressure, pH 6.5. The solubility was evaluated visually. If the test peptide was dissolved completely in the first aliquot of saline solution, its solubility was calculated, and claimed a larger value than the calculated amount. Thus, if 3.5 mg of the test peptide is dissolved immediately in the first 25 μl of the aliquot of 0.5 N. salt solution, its solubility as set >140 mg/ml similarly, if 1.7 mg of the test peptide is not dissolved in 1.7 ml of 0.5 N. salt solution, its solubility set as <1 mg/ml the results of the evaluation of the different solubility of the tested peptides own the Ana in the list in Table VII.

The solubility of various peptides in half-normal saline solution can also be determined using one of the following two semi-quantitative methods - Method 1 and Method 2. Method 1 can be used for those peptides whose solubility in half-normal saline solution is less than about 10 mg/ml, whereas Method 2 can be used for peptides which are soluble in half-normal saline at a concentration of more than about 30 mg/ml

Method 1

Each peptide was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 1 mg/ml. Aliquot of this solution is diluted five-fold in half-normal saline to obtain a solution of 0.2 mg/ml of the resulting peptide solution was subjected to HPLC analysis using a C18/5 microbalance. Buffers for elution consist of a 0.1% solution triperoxonane acid (TFA) in water (buffer A) and 0.1% solution of TFA in 100% acetonitrile (buffer B). Peptides diluted in the gradient buffer is In the range from 5% to 45% within 20 minutes the Area under the curve (AUC) for each peptide peak can be thus defined. This AUC value can be used as a standard (AUC Std)to determine the concentration of each of the corresponding peptide in the supernatant of a saturated solution.

A saturated solution of each peptide can be prepared by adding appropriate to the number of peptide to 1 ml of half-normal saline solution to obtain a turbid suspension. The latter then centrifuged at 10000g for about 10 minutes the Supernatant is then subjected to HPLC analysis. The supernatant is then diluted in half-normal saline solution, and if necessary, bred before HPLC analysis. AUC (AUC-Sat) value obtained from this analysis are used to determine the concentration of peptide in a saturated solution according to the following formula:

The concentration of peptide in a saturated solution=AUC-Std × 0,2/AUC-Sat.

Method 2

This method can provide an assessment of the solubility of the peptide in half-normal saline. The method consists of adding the weighted number (denoted as "XX" milligrams) peptide to 1 ml of half-normal saline to dissolve. The results are presented as >XX mg/ml, where XX is the number of peptide added to half-normal saline solution.

Table VII - Peptide solubility at 20°C in 0.5 N. saline, pH 6.5
No.The solubility of 0.5 N. salt solution (mg/ml)
myr-peptide 1<5,0
Ac-peptide 1>125
myr-peptide 232 >15
myr-peptide 11>2,0
myr-peptide 37>2,0
myr-peptide 79>2,0
myr-peptide 238>2,0
myr-peptide 233>3,0
myr-peptide 234>3,0
myr-peptide 235>60
peptide 79>60
myr-peptide 79-NH2<1,0
myr-peptide 237<1,0
myr-peptide 237-NH2<1,0
peptide 237>80
myr-peptide 234-NH2<2,0
Ac-peptide 79-NH2>60
Ac-peptide 79>100
Ac-peptide 239>50
Ac-peptide 240N/A
Ac-peptide 241>50
myr-peptide 106>10
peptide 106>70
myr-peptide 106-NH2<10
myr-peptide 236<10
myr-peptide 236-NH2<10
peptide 106-NH2>100
Ac-peptide 106>100
cyclic-peptide 106>150
Ac-peptide 242>100
Ac-peptide 243>50
Ac-peptide 236>80
Ac-peptide 244>120
Ac-peptide 245>100
Ac-peptide 247>100
Ac-peptide 248>100
Ac-peptide 249<1
myr-peptide 121<1,0
Ac-peptide 121>20
myr-peptide 137<1,0
myr-peptide 137-NH2N/A
Ac-peptide 250N/A
Ac-peptide 137>200
myr-peptide 15>80
myr-peptide 45>80
myr-peptide 91<20
myr-peptide 153<10
myr-peptide 143<1,0
Ac-peptide 143>230
myr-peptide 179<1,0
Ac-peptide 179>150
myr-peptide 219<1,0
Ac-peptide 219>200
Ac-peptide 219-NH2>200
Ac-peptide 251 >200
Ac-peptide 93-NH2>90
Ac-peptide 108-NH2>150
Ac-peptide 124-NH2>100
Ac-peptide 141-NH2>200
Ac-peptide 159-NH2>200
Ac-peptide 246<30
Ac-peptide 252<30

Example 3 Stability of the tested peptides in biological fluids

Various test peptides were analyzed to determine the sensitivity of the test peptides to proteolysis in biological fluids such as human plasma, BAL fluid of man and the mucus of patients with CF. In addition, for samples that detected the decrease of concentration, has made kinetic analyses of the first order in order to determine the half-life of the tested peptides. The samples were analyzed or the date of receipt or stored at -20°C and analyzed within the next two days.

I. Collection and processing of biological fluids

A. Plasma of a person:

Samples of fresh human blood was collected in vacuo the main tubes for collecting blood with citrate buffer (in the absence of EDTA or heparin). Red blood cells (RBC) were removed by centrifugation of blood at 4000g for 10 min at 4°C. Aliquots of plasma (0.9 ml) was then mixed with 0.1 ml of the test peptide concentration of 0.5 mg/ml in buffer 75 mm sodium acetate at pH 7.0 and incubated in a water bath, keeping the temperature at 37°C. Identical aliquots of 10 µl were taken over time intervals of 5, 15, 30, 60 and 180 minutes and immediately "blocked"by adding 990 μl of a solution consisting of 50% acetonitrile + 50% water containing of 0.2% formic acid. The samples then were subjected to analysis of liquid chromatography-mass spectrometry (IHMS).

Century BALF person:

The BALF samples collected from COPD patients were obtained and frozen. The BALF samples were thawed, mixed together and centrifuged at 10000g for 10 min at 4-8°C. the Supernatant (0.9 ml) was mixed with 0.1 ml of the test peptide concentration of 0.5 mg/ml in buffer 75 mm sodium acetate at pH 7.0, and then processional and analyzed as described above.

C. Mucus person with CF:

Frozen slime (saliva) of patients with CF were thawed and mixed with 2 volumes of buffer 75 mm sodium acetate at pH 7.0 using a glass tissue homogenizer, centrifuged at 10000g for 10 min at 4-8°C. the Precipitate resuspendable in 1 volume of acetate buffer and centrifuged. The two supernatants were combined and is used, as follows. The supernatant (450 µl) was mixed with 50 μl of the peptide solution with a concentration of 0.5 mg/ml in buffer 75 mm sodium acetate at pH 7.0, and then processional and analyzed as described above.

II. Concentration analysis

All samples were analyzed using LC/MS/MS (MDS/SCIEX API 4000 Model). Chromatography was performed using a Phenomenex Luna C18 column, mass spectrometry was performed using an ionization elektrorazpredelenie positive ions.

A. analysis of the peptides Ac-PEPTIDE 79-NH2, PEPTIDE 106-NH2Ac-PEPTIDE 106 in plasma and BAL fluid, and analysis of peptide myr-PEPTIDE 106, PEPTIDE 106, PEPTIDE 106-NH2AC-PEPTIDE 106 and cyc-PEPTIDE 106 (cyclic peptide) in plasma and mucus person with CF

Calibration standards were analyzed using IHMS at concentrations 0,100 µg/ml, 1.00 μg/ml, 10.0 μg/ml and 100 μg/ml for each peptide, with the exception of calibration standards for peptide myr-PEPTIDE 106, PEPTIDE 106, PEPTIDE 106-NH2cyclic peptide cyc-PEPTIDE 106 in the mucus of the person who analyzed at concentrations 0,100 µg/ml, 1.00 μg/ml, 10.0 μg/ml and 75 μg/ml Standards were prepared in 1% BALF human plasma or in the mucus dissolved in a mixture of 50/50 water/acetonitrile and containing 0.2% formic acid. The magnitude of the sensitivity of the device for each set of standards was picked up, COI is lsua inversely proportional to the square of the concentration of the linearized line of least squares. Concentrations of samples were determined using the angle and the intersection of this line. Concentrations abroad limit calibration was determined by extrapolation of the calibration curve.

C. Analysis of peptide myr-PEPTIDE 234-NH2, myr-PEPTIDE 234, PEPTIDE 106 and myr-PEPTIDE 106 in plasma and BALF

Identical to the calibration standards were analyzed at a concentration of 50 μg/ml of each peptide. The magnitude of the sensitivity of the device for each set of standards was picked up by intersection with 0, using unweighted linear line of least squares. Concentrations of samples were determined using the slope and the intersection of this line.

C. Analysis of peptides PEPTIDE 237, myr-PEPTIDE 106-NH2, myr-PEPTIDE 236 and myr-PEPTIDE 236-NH2in plasma and BALF

For analysis of peptide concentration in plasma, is identical to the calibration standards were analyzed at concentrations of 25 μg/ml and 50 μg/ml of each peptide. The magnitude of the sensitivity of the device for each set of standards was picked up by intersection with 0, using unweighted linear line of least squares. Concentrations of samples were determined using the slope and the intersection of this line. Concentrations abroad limit calibration was determined by extrapolation of the calibration curve.

For analysis of pepti the Noy concentration in the BALF samples one calibration standard concentration of 50 μg/ml, cooked in a mixture of BALF, water, acetonitrile and formic acid, were analyzed with each set of samples.

III. Kinetic profiles

The kinetic profiles of the first order was determined using the program Watson for all samples, which showed a marked decrease in concentration. Watson leads in accordance logarithmically transformed data with the line of least squares to determine parameters such asmax(maximum concentration), the intersection, the coefficient of kinetic, tilt and T1/2(half-life). The obtained kinetic parameters were based on linear selection, and not on the actual values of concentration.

The zero point of the calibration standards in the solution were not included in the kinetic profiles. But the real kinetics of the first order does not require the inclusion of data on the zero point in order to accurately describe the change in concentration as a function of time.

The half-life periods of the tested peptides in plasma (Plasma t1/2), BALF person (BALF t1/2and CF mucus person (Slime t1/2) listed in Table VIII.

Table VIII
ID Peptide #Plasma t1/2/sub>
(hours)
(1thexp.)
Plasma t1/2
(hours)
(2thexp.)
BALF t1/2
(hours)
(1thexp.)
BALF t1/2
(hours)
(2thexp.)
Slime t1/2
(hours)
(1thexp.)
Slime t1/2
(hours)
(2thexp.)
myr-peptide 234-NH23,82--1,70------
myr-peptide 234*--1,08------
peptide 1060,280,151,88--0,230,37
myr-peptide 1061,230,901,03--at 9.533,00
peptide 2370,49--**----
myr-peptide 106-NH2*--10,5212,81----
myr-peptide 2360,72--3,552,94----
myr-peptide 236-NH2*--4,164,08----
Ac-peptide 79-NH22,64--*------
peptide 106-NH20,300,134,00-- 0,080,13
Ac-peptide 106*0,031,89--0,550,47
cyclic-peptide 106--1,04----1,102,00
#: myr and Ac are respectively myristoleic and acetyl groups covalently linked to the peptide on the N-terminal site of the amino group; -NH2is a covalently linked amide at the C-terminal carboxyl group of the peptide; seq no and cyc is an abbreviation denoting the sequence number and cyclic, respectively.
--: The experiment was not performed.
*: Not enough experimental data to calculate the half-life.

The peptide designated as myr-peptide 236 is myristoyl-PEPTIDE 236. All other peptides used in the experiment described above.

Example 4 - Effect of MANS peptide for the secretion of mucus in the upper respiratory tract of primates

The aim of the experiment was to determine the ability of the MANS peptide to inhibit reception is the function of mucus in the upper respiratory tract of adults healthy rhesus macaques. Used test is the standard method for evaluating nasal secretory activity, and primates usually provide a good correlation with such activity in humans.

Methods

For this experiment used a 17 healthy young adult male rhesus macaques that do not have any history of rhinitis. None of the monkeys was not sick rhinitis before or after the study. Nasal mucus secretion each monkey was measured in her left nostril before any processing. This value is taken for 100%

mucus secretion. Monkeys then randomly divided into the following 4 groups:

Group 1: Normal saline control (n=3);

Group 2: sodium Acetate, the control solvent (n=4);

Group 3: RNS peptide, negative control (n=5);

Group 4: MANS peptide, the test peptide (n=5).

Saline was placed in the left nostril all 17 monkeys before any processing. The right nostril was treated with 2.0 ml or saline, sodium acetate, RNS peptide or MANS peptide. Thus, each animal was its own control. Nasal lavage was performed on both nostrils of each animal within 1 hour after processing the test compositions. All lavage were immediately frozen at -80°C and analyzed for the content of mucus by using ELISA method.

A. under Test to which notizie:

Normal saline, sterilized by filtration; sodium Acetate, 150 μm, sterilized by filtration; RNS Peptide - 140 μm solution of 150 μm solution, sterilized by filtration, sodium acetate and MANS Peptide - 140 μm solution of 150 μm solution, sterilized by filtration, sodium acetate.

C. Test animals:

Rhesus macaques; number of animals: 17; gender: all healthy males; age: 3-4 years; body weight: 4-7 kg, the average of 5.03 kg; acclimatization period: 7 days; method of identification: a unique tattoo of a 5-digit ID number; history: all animals for 12-24 months before this test was used to study vaccine, namely immunization against meningitis. All animals also received regular immunization against measles and tetanus during infancy.

C. Keeping of animals:

Animal care: conditions correspond to the standard routine procedures, which are based on the "Guide for the care and use of laboratory animals". Food: standard diet rhesus makaka provided daily; water: freely available municipal water delivered through the automatic water supply system; placement: animals were placed individually in proven corrosion-resistant steel cells, identified with a card showing the number of animals, the code test is, floor, animal code; environment: room temperature were measured daily. Temperature limits for the room was in the range of 20-26°C. the Humidity in the room was measured daily, border humidity was in the range of 40-70%. The light cycle was controlled using an automatic timer (12 hours light, 12 hours dark); staff: staff involved were appropriately qualified and trained to work with primates.

Results

The results present evidence that RNS peptide, the buffer is sodium acetate or normal saline has no effect on the secretion of mucus, while almost 75% MANS peptides inhibit the secretion of mucus.

Example 5 Method of cell cultures, tissues to determine the greater of mucin in bronchial epithelial human cells

HBE1 is a virus-transformed bronchial epithelial cell line human, capable of secreting mucin under cultivation in the boundary conditions of air and fluid. HBE1 cells were cultured in the boundary conditions of air and fluid, as described previously (Li et al., J.Biol.Chem., volume 276, pp 40982-40990, 2001). Briefly, HBE1 cells were cultured in the boundary conditions of air and fluid through rasseivaniya the appropriate number of cells in a 12-hole pure culture plates (Costar, Cambrige, MA), which were p is covered with a thin layer of collagen type I, isolated from rat tail (Collaborative Biomedical, Bedford, MA). First, for 5-7 days at a humidity of 95% and 5% CO2in the environment the cells were maintained immersed in the environment as long as the cells will not be placed close to each other. At this time created boundary conditions of air and fluid by removing the apical part of the environment and carried out the power cells of the lateral. After that, the medium was changed daily. Cells were additionally cultured for another 14 days to reach full differentiation. Accumulated mucin on the apical surface of the cells were removed by washing with salt solution in phosphate buffer, pH 7,2. To collect baseline secretion of mucin, the cells were incubated for 30 min in the same medium and collected mucin, secreted into the apical medium, and evaluated its number using ELISA. To determine the hypersecretion of mucin induced secretory agent, inducing mucin, cells were exposed for 30 min with medium containing 0.5 µm of formalparameterlist (PMA), after which the mucin was collected and evaluated its number using ELISA. To determine the inhibition by the test peptide PMA-induced mucin hypersecretion, cells were preincubated in a medium containing 25 or 50 μm of the test peptide for 15 min, followed the incubation with 0.5 µm PMA for 30 minutes Six wells were used for each test peptide and for each control. The mucin, secreted into the apical medium was collected and evaluated its quantity by the sandwich ELISA using specific mucin antibodies conjugated with alkaline phosphatase (MUC5A) (Zymed Laboratories, San Francisco, CA).

Processing of HBE1 cells with 0.5 μm PMA resulted in increased secretion of mucin by 20%. PMA-induced increase in the secretion of mucin blocked 100% of pre-treatment with 25 μm MANS peptide or treatment with 25 μm Ac-peptide No. 106. Ac-peptide No. 219 at a concentration of 25 μm, not only inhibited 100% of PMA-induced secretion of mucin, but also inhibited the secretion of mucin to the level of 20% smaller than in estimulando control environment. Ac-peptide No. 251 had only a minor effect of inhibition only 6% of the PMA-induced increase in the secretion of mucin.

Table IX contains the list of the peptides of the present invention and corresponding amino acid sequences and corresponding SEQ ID NO.

Table IX. Peptides and amino acid sequence
Peptide No.SequenceSequence ID No.
peptide 1 GAQFSKTAAKGEAAAERPGEAAVASEQ ID NO. 1
peptide 2GAQFSKTAAKGEAAAERPGEAAVSEQ ID NO. 2
peptide 3AQFSKTAAKGEAAAERPGEAAVASEQ ID NO. 3
peptide 4GAQFSKTAAKGEAAAERPGEAASEQ ID NO. 4
peptide 5AQFSKTAAKGEAAAERPGEAAVSEQ ID NO. 5
peptide 6QFSKTAAKGEAAAERPGEAAVASEQ ID NO. 6
peptide 7GAQFSKTAAKGEAAAERPGEASEQ ID NO. 7
peptide 8AQFSKTAAKGEAAAERPGEAASEQ ID NO. 8
peptide 9QFSKTAAKGEAAAERPGEAAVSEQ ID NO. 9
peptide 10FSKTAAKGEAAAERPGEAAVASEQ ID NO. 10
peptide 11GAQFSKTAAKGEAAAERPGESEQ ID NO. 11
peptide 12AQFSKTAAKGEAAAERPGEA SEQ ID NO. 12
peptide 13QFSKTAAKGEAAAERPGEAASEQ ID NO. 13
peptide 14FSKTAAKGEAAAERPGEAAVSEQ ID NO. 14
peptide 15SKTAAKGEAAAERPGEAAVASEQ ID NO. 15
peptide 16GAQFSKTAAKGEAAAERPGSEQ ID NO. 16
peptide 17AQFSKTAAKGEAAAERPGESEQ ID NO. 17
peptide 18QFSKTAAKGEAAAERPGEASEQ ID NO. 18
peptide 19FSKTAAKGEAAAERPGEAASEQ ID NO. 19
peptide 20SKTAAKGEAAAERPGEAAVSEQ ID NO. 20
peptide 21KTAAKGEAAAERPGEAAVASEQ ID NO. 21
peptide 22GAQFSKTAAKGEAAAERPSEQ ID NO. 22
peptide 23AQFSKTAAKGEAAAERPGSEQ ID NO. 23

peptide 24QFSKTAAKGEAAAERPGESEQ ID NO. 24
peptide 25FSKTAAKGEAAAERPGEASEQ ID NO. 25
peptide 26SKTAAKGEAAAERPGEAASEQ ID NO. 26
peptide 27KTAAKGEAAAERPGEAAVSEQ ID NO. 27
peptide 28TAAKGEAAAERPGEAAVASEQ ID NO. 28
peptide 29GAQFSKTAAKGEAAAERSEQ ID NO. 29
peptide 30AQFSKTAAKGEAAAERPSEQ ID NO. 30
peptide 31QFSKTAAKGEAAAERPGSEQ ID NO. 31
peptide 32FSKTAAKGEAAAERPGESEQ ID NO. 32
peptide 33SKTAAKGEAAAERPGEASEQ ID NO. 33
peptide 34KTAAKGEAAAERPGEAASEQ ID NO. 34
peptide 35TAAKGEAAAERPGEAV SEQ ID NO. 35
peptide 36AAKGEAAAERPGEAAVASEQ ID NO. 36
peptide 37GAQFSKTAAKGEAAAESEQ ID NO. 37
peptide 38AQFSKTAAKGEAAAERSEQ ID NO. 38
peptide 39QFSKTAAKGEAAAERPSEQ ID NO. 39
peptide 40FSKTAAKGEAAAERPGSEQ ID NO. 40
peptide 41SKTAAKGEAAAERPGESEQ ID NO. 41
peptide 42KTAAKGEAAAERPGEASEQ ID NO. 42
peptide 43TAAKGEAAAERPGEAASEQ ID NO. 43
peptide 44AAKGEAAAERPGEAAVSEQ ID NO. 44
peptide 45AKGEAAAERPGEAAVASEQ ID NO. 45
peptide 46GAQFSKTAAKGEAAASEQ ID NO. 46
peptide 47 AQFSKTAAKGEAAAESEQ ID NO. 47
peptide 48QFSKTAAKGEAAAERSEQ ID NO. 48
peptide 49FSKTAAKGEAAAERPSEQ ID NO. 49

peptide 50SKTAAKGEAAAERPGSEQ ID NO. 50
peptide 51KTAAKGEAAAERPGESEQ ID NO. 51
peptide 52TAAKGEAAAERPGEASEQ ID NO. 52
peptide 53AAKGEAAAERPGEAASEQ ID NO. 53
peptide 54AKGEAAAERPGEAAVSEQ ID NO. 54
peptide 55KGEAAAERPGEAAVASEQ ID NO. 55
peptide 56GAQFSKTAAKGEAASEQ ID NO. 56
peptide 57AQFSKTAAKGEAAASEQ ID NO. 57
peptide 58QFSKTAAKGEAAAEEQ ID NO. 58
peptide 59FSKTAAKGEAAAERSEQ ID NO. 59
peptide 60SKTAAKGEAAAERPSEQ ID NO. 60
peptide 61KTAAKGEAAAERPGSEQ ID NO. 61
peptide 62TAAKGEAAAERPGESEQ ID NO. 62
peptide 63AAKGEAAAERPGEASEQ ID NO. 63
peptide 64AKGEAAAERPGEAASEQ ID NO. 64
peptide 65KGEAAAERPGEAAVSEQ ID NO. 65
peptide 66GEAAAERPGEAAVASEQ ID NO. 66
peptide 67GAQFSKTAAKGEASEQ ID NO. 67
peptide 68AQFSKTAAKGEAASEQ ID NO. 68
peptide 69QFSKTAAKGEAAASEQ ID NO. 69
peptide 70FSKTAAKGEAAAE SEQ ID NO. 70
peptide 71SKTAAKGEAAAERSEQ ID NO. 71
peptide 72KTAAKGEAAAERPSEQ ID NO. 72
peptide 73TAAKGEAAAERPGSEQ ID NO. 73
peptide 74AAKGEAAAERPGESEQ ID NO. 74
peptide 75AKGEAAAERPGEASEQ ID NO. 75

peptide 76KGEAAAERPGEAASEQ ID NO. 76
peptide 77GEAAAERPGEAAVSEQ ID NO. 77
peptide 78EAAAERPGEAAVASEQ ID NO. 78
peptide 79GAQFSKTAAKGESEQ ID NO. 79
peptide 80AQFSKTAAKGEASEQ ID NO. 80
peptide 81QFSKTAAKGEAASEQ ID NO. 81
peptide 2 FSKTAAKGEAAASEQ ID NO. 82
peptide 83SKTAAKGEAAAESEQ ID NO. 83
peptide 84KTAAKGEAAAERSEQ ID NO. 84
peptide 85TAAKGEAAAERPSEQ ID NO. 85
peptide 86AAKGEAAAERPGSEQ ID NO. 86
peptide 87AKGEAAAERPGESEQ ID NO. 87
peptide 88KGEAAAERPGEASEQ ID NO. 88
peptide 89GEAAAERPGEAASEQ ID NO. 89
peptide 90EAAAERPGEAAVSEQ ID NO. 90
peptide 91AAAERPGEAAVASEQ ID NO. 91
peptide 92GAQFSKTAAKGSEQ ID NO. 92
peptide 93AQFSKTAAKGESEQ ID NO. 93
pepti is 94 QFSKTAAKGEASEQ ID NO. 94
peptide 95FSKTAAKGEAASEQ ID NO. 95
peptide 96SKTAAKGEAAASEQ ID NO. 96
peptide 97KTAAKGEAAAESEQ ID NO. 97
peptide 98TAAKGEAAAERSEQ ID NO. 98
peptide 99AAKGEAAAERPSEQ ID NO. 99
peptide 100AKGEAAAERPGSEQ ID NO. 100
peptide 101KGEAAAERPGESEQ ID NO. 101

peptide 102GEAAAERPGEASEQ ID NO. 102
peptide 103EAAAERPGEAASEQ ID NO. 103
peptide 104AAAERPGEAAVSEQ ID NO. 104
peptide 105AAERPGEAAVA SEQ ID NO. 105
peptide 106GAQFSKTAAKSEQ ID NO. 106
peptide 107AQFSKTAAKGSEQ ID NO. 107
peptide 108QFSKTAAKGESEQ ID NO. 108
peptide 109FSKTAAKGEASEQ ID NO. 109
peptide 110SKTAAKGEAASEQ ID NO. 110
peptide 111KTAAKGEAAASEQ ID NO. 111
peptide 112TAAKGEAAAESEQ ID NO. 112
peptide 113AAKGEAAAERSEQ ID NO. 113
peptide 114AKGEAAAERPSEQ ID NO. 114
peptide 115KGEAAAERPGSEQ ID NO. 115
peptide 116GEAAAERPGESEQ ID NO. 116
peptide 117EAAAERPGEA SEQ ID NO. 117
peptide 118AAAERPGEAASEQ ID NO. 118
peptide 119AAERPGEAAVSEQ ID NO. 119
peptide 120AERPGEAAVASEQ ID NO. 120
peptide 121GAQFSKTAASEQ ID NO. 121
peptide 122AQFSKTAAKSEQ ID NO. 122
peptide 123QFSKTAAKGSEQ ID NO. 123
peptide 124FSKTAAKGESEQ ID NO. 124
peptide 125SKTAAKGEASEQ ID NO. 125
peptide 126KTAAKGEAASEQ ID NO. 126
peptide 127TAAKGEAAASEQ ID NO. 127

peptide 128AAKGEAAAESEQ ID NO. 128
peptide 129 AKGEAAAERSEQ ID NO. 129
peptide 130KGEAAAERPSEQ ID NO. 130
peptide 131GEAAAERPGSEQ ID NO. 131
peptide 132EAAAERPGESEQ ID NO. 132
peptide 133AAAERPGEASEQ ID NO. 133
peptide 134AAERPGEAASEQ ID NO. 134
peptide 135AERPGEAAVSEQ ID NO. 135
peptide 136ERPGEAAVASEQ ID NO. 136
peptide 137GAQFSKTASEQ ID NO. 137
peptide 138AQFSKTAASEQ ID NO. 138
peptide 139QFSKTAAKSEQ ID NO. 139
peptide 140FSKTAAKGSEQ ID NO. 140
peptide 141SKTAKGE SEQ ID NO. 141
peptide 142KTAAKGEASEQ ID NO. 142
peptide 143TAAKGEAASEQ ID NO. 143
peptide 144AAKGEAAASEQ ID NO. 144
peptide 145AKGEAAAESEQ ID NO. 145
peptide 146KGEAAAERSEQ ID NO. 146
peptide 147GEAAAERPSEQ ID NO. 147
peptide 148EAAAERPGSEQ ID NO. 148
peptide 149AAAERPGESEQ ID NO. 149
peptide 150AAERPGEASEQ ID NO. 150
peptide 151AERPGEAASEQ ID NO. 151
peptide 152ERPGEAAVSEQ ID NO. 152
peptide 153RPGEAAVA SEQ ID NO. 153

peptide 154GAQFSKTSEQ ID NO. 154
peptide 155AQFSKTASEQ ID NO. 155
peptide 156QFSKTAASEQ ID NO. 156
peptide 157FSKTAAKSEQ ID NO. 157
peptide 158SKTAAKGSEQ ID NO. 158
peptide 159KTAAKGESEQ ID NO. 159
peptide 160TAAKGEASEQ ID NO. 160
peptide 161AAKGEAASEQ ID NO. 161
peptide 162AKGEAAASEQ ID NO. 162
peptide 163KGEAAAESEQ ID NO. 163
peptide 164GEAAAERSEQ ID NO. 164
peptide 165 EAAAERPSEQ ID NO. 165
peptide 166AAAERPGSEQ ID NO. 166
peptide 167AAERPGESEQ ID NO. 167
peptide 168AERPGEASEQ ID NO. 168
peptide 169ERPGEAASEQ ID NO. 169
peptide 170RPGEAAVSEQ ID NO. 170
peptide 171PGEAAVASEQ ID NO. 171
peptide 172GAQFSKSEQ ID NO. 172
peptide 173AQFSKTSEQ ID NO. 173
peptide 174QFSKTASEQ ID NO. 174
peptide 175FSKTAASEQ ID NO. 175
peptide 176SKTAAKSEQ ID NO. 176
peptide 177KTAAKGpeptide 178TAAKGESEQ ID NO. 178
peptide 179AAKGEASEQ ID NO. 179

peptide 180AKGEAASEQ ID NO. 180
peptide 181KGEAAASEQ ID NO. 181
peptide 182GEAAAESEQ ID NO. 182
peptide 183EAAAERSEQ ID NO. 183
peptide 184AAAERPSEQ ID NO. 184
peptide 185AAERPGSEQ ID NO. 185
peptide 186AERPGESEQ ID NO. 186
peptide 187ERPGEASEQ ID NO. 187
peptide 188RPGEAASEQ ID NO. 188
peptide 189 SEQ ID NO. 189
peptide 190GEAAVASEQ ID NO. 190
peptide 191GAQFSSEQ ID NO. 191
peptide 192AQFSKSEQ ID NO. 192
peptide 193QFSKTSEQ ID NO. 193
peptide 194FSKTASEQ ID NO. 194
peptide 195SKTAASEQ ID NO. 195
peptide 196KTAAKSEQ ID NO. 196
peptide 197TAAKGSEQ ID NO. 197
peptide 198AAKGESEQ ID NO. 198
peptide 199AKGEASEQ ID NO. 199
peptide 200KGEAASEQ ID NO. 200
peptide 201GEAAASE ID NO. 201
peptide 202EAAAESEQ ID NO. 202
peptide 203AAAERSEQ ID NO. 203
peptide 204AAERPSEQ ID NO. 204
peptide 205AERPGSEQ ID NO. 205

peptide 206ERPGESEQ ID NO. 206
peptide 207RPGEASEQ ID NO. 207
peptide 208PGEAASEQ ID NO. 208
peptide 209GEAAVSEQ ID NO. 209
peptide 210EAAVASEQ ID NO. 210
peptide 211GAQFSEQ ID NO. 211
peptide 212AQFSSEQ ID NO. 212
peptide 213QFSK SEQ ID NO. 213
peptide 214FSKTSEQ ID NO. 214
peptide 215SKTASEQ ID NO. 215
peptide 216KTAASEQ ID NO. 216
peptide 217TAAKSEQ ID NO. 217
peptide 218AAKGSEQ ID NO. 218
peptide 219AKGESEQ ID NO. 219
peptide 220KGEASEQ ID NO. 220
peptide 221GEAASEQ ID NO. 221
peptide 222EAAASEQ ID NO. 222
peptide 223AAAESEQ ID NO. 223
peptide 224AAERSEQ ID NO. 224
peptide 225AERPSEQ ID NO. 225
ERPGSEQ ID NO. 226
peptide 227RPGESEQ ID NO. 227
peptide 228PGEASEQ ID NO. 228
peptide 229GEAASEQ ID NO. 229
peptide 230EAAVSEQ ID NO. 230
peptide 231AAVASEQ ID NO. 231

peptide 232GTAPAAEGAGAEVKRASAEAKQAFSEQ ID NO. 232
peptide 233GKQFSKTAAKGESEQ ID NO. 233
peptide 234GAQFSKTKAKGESEQ ID NO. 234
peptide 235GKQFSKTKAKGESEQ ID NO. 235
peptide 236GAQASKTAAKSEQ ID NO. 236
peptide 237GAQASKTAAKGE SEQ ID NO. 237
peptide 238GAEFSKTAAKGESEQ ID NO. 238
peptide 239GAQFSKTAAAGESEQ ID NO. 239
peptide 240GAQFSKTAAKAESEQ ID NO. 240
peptide 241GAQFSKTAAKGASEQ ID NO. 241
peptide 242AAQFSKTAAKSEQ ID NO. 242
peptide 243GAAFSKTAAKSEQ ID NO. 243
peptide 244GAQFAKTAAKSEQ ID NO. 244
peptide 245GAQFSATAAKSEQ ID NO. 245
peptide 246KAATKSFQAGSEQ ID NO. 246
peptide 247GAQFSKAAAKSEQ ID NO. 247
peptide 248GAQFSKTAAASEQ ID NO. 248
peptide 249GAQFSATAAA SEQ ID NO. 249
peptide 250GAQASKTASEQ ID NO. 250
peptide 251AAGESEQ ID NO. 251
peptide 252GKASQFAKTASEQ ID NO. 252

While the invention is described in detail and with reference to specific embodiments, it will be no doubt to anyone skilled in the art that various changes and modifications can be made without departing from the scope of the essence and scope of the invention.

1. Dedicated terminal modified peptide that contains the amino acid sequence selected from the group consisting of:
(a) sequence of 24 amino acids having the sequence GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO: 1)or amino acid sequences that have at least one amino acid substitution in the specified reference sequence selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or substitution of Q on E;
(b) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO: 1)or amino acid sequences that have at least one the amino acid replacement in the specified reference sequence, selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or substitution of Q on E, and the amino acid sequence of the peptide begins with the N-terminal amino acids of the reference sequence; and
(C) amino acid sequence having from 4 to 23 consecutive amino acids of the reference sequence GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO: 1)or amino acid sequences that have at least one amino acid substitution in the specified reference sequence selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or substitution of Q on E, and the amino acid sequence of the peptide begins with amino acid position 2 to amino acid position 21 of the reference sequence (SEQ ID NO: 1); and
(i) the C-terminal carboxyl group of the C-terminal amino acids of peptide liderovna and N-terminal amino group of the N-terminal amino acids of peptide azetilirovanna, or
(ii) C-terminal carboxyl group of the C-terminal amino acids of peptide liderovna and N-terminal amino group of the N-terminal amino acids of the peptide is not chemically modified, or
(iii) C-terminal carboxyl group of the C-terminal amino acids of the peptide is not chemically modified and the N-terminal amino group of the N-terminal amino acids of peptide azetilirovanna, and when serial amino acid is inost peptide represents (a) or (b), N-terminal amino acid is not monitorowania; and
when the amino acid sequence of (b) and (C) consists of 4 amino acids, the sequence contains SEQ ID NO: 211-221 or 223-230;
where the peptide has the effect of inhibiting mucin hypersecretion when administered to a mammal in number, the inhibitory hypersecretion of mucin.

2. The peptide according to claim 1, where the amino acid sequence of the peptide begins with the N-terminal amino acids of the reference sequence.

3. The peptide according to claim 1, where the amino acid sequence of the peptide begins with amino acid position 2 to amino acid position 21 of the reference sequence.

4. The peptide according to claim 1, where the amino acid sequence of the peptide ends C-terminal amino acid of the reference sequence.

5. The peptide according to claim 1, where the amino acid sequence of the peptide comprises sequentially located the remains of AKGE (SEQ ID NO: 219) reference sequence.

6. The peptide according to any one of claims 1 to 5, where amino acid sequence of the peptide consists of 4-16 consecutive amino acids of the reference sequence or amino acid sequence that has at least one amino acid substitution in the specified reference sequence selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or is amany Q on E.

7. The peptide according to any one of claims 1 to 5, where amino acid sequence of the peptide consists of from 8 to 14 consecutive amino acids of the reference sequence or amino acid sequence that has at least one amino acid substitution in the specified reference sequence selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or substitution of Q on E.

8. The peptide according to any one of claims 1 to 5, where amino acid sequence of the peptide consists of 10-12 consecutive amino acids of the reference sequence or amino acid sequence that has at least one amino acid substitution in the specified reference sequence selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or substitution of Q on E.

9. The peptide according to any one of claims 1 to 5, where one or more additional amino acids of the peptide is independently alkylated, lidirovali, etilirovany or tarifitsirovana.

10. The peptide according to any one of claims 1 to 5, where N-terminal amino acid of the peptide azetilirovanna N-terminal amino group.

11. The peptide according to claim 1, which is selected from the group consisting of acetyl-PEPTIDE 106, acetyl-PEPTIDE 103-NH2, PEPTIDE 106-NH2, PEPTIDE 234-NH2and acetyl-PEPTIDE 219.

12. The peptide according to claim 1, which is selected from the group consisting of SEQ ID NO: 15, SEQ ID NO: 45, SEQ ID NO: 91, SEQ ID N: 153, SEQ ID NO: 79, SEQ ID NO: 233, SEQ ID NO: 234 and SEQ ID NO: 235.

13. The peptide according to claim 1, which is selected from the group consisting of SEQ ID NO: 233-245 and SEQ ID nos: 247-251.

14. The peptide according to any one of claims 1 to 5, 11, 12 or 13, where the sequence contains the amino acid sequence of the reference sequence that has at least one amino acid substitution selected from the group consisting of replacement And To replace F, K, G, Q, S, T and/or E in A; or substitution of Q on E.

15. The peptide according to claim 1, having the amino acid sequence GAQFSKTAAKGEAAAERPGEAAVA (SEQ ID NO: 1), where N-terminal amino acid of the peptide azetilirovanna.

16. The peptide according to any one of claims 1 to 5 or 12-13, where the C-terminal amino acid of the peptide liderovna on the C-terminal carboxyl group of the amino group selected from the group consisting of: ammonia, C1-C24 aliphatic alkylamino, hydroxyl-substituted C2-C24 aliphatic alkylamines, linear 2-(C1-C24 aliphatic alkyl)acetylamino or omega-methoxy-poly(ethyleneoxy)n-etilenovomu, where n is from 0 to 10.

17. The peptide according to any one of claims 1 to 5 or 12-13, which exhibits at least one of the properties: (a) greater effect of inhibiting mucin hypersecretion in a mammal than showing the sequence of SEQ ID NO: 1, in which the N-terminal amino acid monitorowania, with the introduction of the specified mammal in equal concentrations, or (b) high solubility in the ode, than the sequence SEQ ID NO: 1, in which the N-terminal amino acid monitorowania, at equal concentrations in the same liquid.

18. The pharmaceutical composition inhibiting the hypersecretion of mucin containing a therapeutically effective amount of a peptide according to any one of claims 1 to 17 and, optionally, a pharmaceutically acceptable carrier and/or diluent.

19. The peptide according to any one of claims 1 to 5, 11-13, 15 for use as drugs having activity of inhibiting the hypersecretion of mucin.

20. The peptide according to any one of claims 1 to 5, 11-13, 15 for use in the treatment of hypersecretion of mucin.

21. The peptide according to claim 20, where the hypersecretion of mucin is a symptom of lung disease.

22. The peptide according to item 21, where pulmonary disease is a asthma, chronic bronchitis, COPD, cystic fibrosis or rhinitis.

23. The use of the peptide according to any one of claims 1 to 17 for the manufacture of a medicinal product for the treatment of hypersecretion of mucin.

24. The application of item 23, where hypersecretion of mucin is a symptom of lung disease.

25. The application of paragraph 24, where pulmonary disease is a asthma, chronic bronchitis, COPD, cystic fibrosis or rhinitis.

26. Method of inhibiting mucin hypersecretion in a mammal, comprising the administration to a mammal inhibits mucin hypersecretion number FA the pharmaceutical composition according p, which inhibits the hypersecretion of mucin.

27. The method according to p where hypersecretion of mucin is a symptom of lung disease.

28. The method according to item 27, where pulmonary disease is a asthma, chronic bronchitis, COPD, cystic fibrosis, or rhinitis.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention discloses a complementary peptide having at least a sequence presented in the description and complementary to a primary immunogenic acetylcholine receptor region involved in myasthenia gravis, having at least the sequence SEQ ID NO:1 with tryptophan in position 8 and bearing at least one optionally substituted hydrocarbon group. There is disclosed therapeutic composition for treating myasthenia gravis based on the complementary peptide and applied for preparing a vaccine used in therapeutic and preventive treatment of myasthenia gravis in mammals. There are described methods of producing the complementary peptide and a drug and a method of treating myasthenia gravis in mammals with using the therapeutic composition.

EFFECT: invention provides the peptide showing an antigen behaviour and which can be used in treating MG in mammals, particularly dogs and humans.

22 cl, 11 dwg, 5 tbl, 3 ex

FIELD: medicine.

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22 cl, 12 ex, 3 dwg, 2 tbl

FIELD: chemistry; biochemistry.

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8 cl, 4 dwg, 1 tbl, 5 ex

FIELD: biotechnology, medicine, oncology, peptides.

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2 dwg, 2 ex

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25 cl, 15 dwg, 6 ex

FIELD: medicine, pharmaceutics.

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30 tbl, 14 ex

FIELD: medicine.

SUBSTANCE: invention relates to compounds of formula

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27 cl, 1 tbl, 5 ex

FIELD: medicine, pharmaceutics.

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4 dwg, 9 tbl, 15 ex

FIELD: medicine, pharmaceutics.

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6 cl, 3 tbl, 2 ex

FIELD: chemistry.

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13 cl, 22 tbl, 281 ex

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5 dwg, 1 tbl, 4 ex

FIELD: chemistry.

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5 ex

FIELD: medicine.

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4 dwg, 2 ex

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25 cl, 15 dwg, 6 ex

FIELD: medicine.

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27 cl, 41 dwg, 35 ex

New application // 2416426

FIELD: medicine, pharmaceutics.

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EFFECT: creating the drug allied with a class of proteins naturally formed in an organism.

32 cl, 6 ex, 3 dwg

FIELD: chemistry.

SUBSTANCE: method of producing iron-containing succinylated casein involves the following steps: a) reaction of casein with at least one succinylation agent to form an aqueous suspension of succinylated casein, where step (a) involves the following steps: (a1) suspension of casein in water, (a2) if necessary, bringing pH of the aqueous suspension to at least 6, (a3) addition of at least one succinylation agent while maintaining pH of at least 6 by adding at least one base, (a4) precipitation of the obtained succinylated casein after adding the succinylation agent by bringing pH to approximately 2-7 to obtain an aqueous suspension of succinylated casein, having pH of approximately 2-7, b) reaction of the aqueous suspension of succinylated casein obtained at step (a) with at least one iron salt to form iron-containing succinylated casein, where step (b) involves addition of at least one iron salt to the aqueous suspension of succinylated casein while keeping pH of casein equal to at least 2 by adding at least one base to obtain iron-containing succinylated casein.

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12 cl, 5 ex, 1 tbl

FIELD: chemistry; biochemistry.

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22 cl, 8 dwg, 1 ex

FIELD: medicine.

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EFFECT: higher anticancer activity of the compounds.

3 cl, 6 dwg, 4 ex

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