Novel conjugates of montelukast and peptides

Abstract

Peptide-containing compounds and positional isomers, stereoisomers, and pharmaceutically or cosmetically acceptable salts of the peptide-containing compounds are provided, wherein the peptide-containing compounds include a peptide component that is an amino acid sequence of 2 to 45 amino acids, and the peptide component is covalently attached to one or more compounds of formula (I). [Formula 1] (In the formula, R 1 is selected from the group consisting of -C(CH3)2OH, -COCH3, -C(CH3)=CH2, and -C(CH3)2H, and n is 0, 1, or 2. The compound of formula I is preferably montelukast, montelukast styrene, or hydrogenated montelukast styrene.Peptide-containing compounds are particularly useful for treating inflammation, including wounds, hemorrhoids, burns, psoriasis, acne, atopic dermatitis, allergic rhinitis, allergic conjunctivitis, chronic obstructive pulmonary disease, and inflammatory bowel disease (such as ulcerative colitis).Peptide-containing compounds are also useful for treating idiopathic pulmonary fibrosis.

Description

[Technical Field]

[0001] The present invention relates to novel conjugates of known anti-inflammatory compounds covalently attached to specific peptide sequences, the use of such novel conjugates in human medicine, and pharmaceutical compositions containing them. Specifically, the present invention relates to the use of these conjugates and compositions in the treatment of inflammation. [Background technology]

[0002] Inflammation is typically characterized as a localized tissue response to invasion by, for example, microorganisms, specific antigens, damaged cells, or physical and / or chemical agents. The inflammatory response is usually a protective mechanism that destroys, weakens, or sequesters both the harmful substance and the injured tissue, and initiates tissue healing.

[0003] Inflammation can result from physical trauma, infection, certain chronic diseases (e.g., autoimmune diseases such as psoriasis and rheumatoid arthritis), and / or a chemical and / or physiological response to an external stimulus (e.g., part of an allergic reaction). A complex series of events can be involved in which inflammatory mediators increase blood flow and local vascular dilation, resulting in redness and heat, exudation of fluid, often resulting in local swelling, leukocyte migration to the inflamed area, and pain.

[0004] Many pathologies / disorders are characterized by and / or caused by abnormal tissue damage inflammation.These pathologies are typically characterized by the activation of immune defense mechanisms, resulting in more harmful effects than beneficial effects for the host, and are generally associated with varying degrees of tissue redness or hyperemia, swelling, hyperthermia, pain, itching, cell death, tissue destruction, cell proliferation, and / or loss of function.Examples include inflammatory bowel disease, rheumatoid arthritis, multiple sclerosis, psoriasis, glomerulonephritis, and transplant rejection.

[0005] Typically, a complex series of events leads to inflammatory changes, including increased blood flow due to local vascular dilation resulting in redness and heat, extravasation of leukocytes and plasma often resulting in local swelling, activation of sensory nerves (resulting in pain in some tissues), and loss of function. These inflammatory changes are caused by a cascade of cellular and biochemical events involving cells such as neutrophils, monocytes, macrophages, and lymphocytes, along with inflammatory mediators such as vasoactive amines, cytokines, complement factors, and reactive oxygen species.

[0006] In particular, inflammation plays an important role in the wound healing process.Therefore, wounds and burns can be classified as inflammation-related pathologies.The conventional thinking in the art is that anti-inflammatory drugs should not be applied directly to open wounds, because it is harmful to the progress of wound healing.

[0007] Montelukast is an orally active nonsteroidal immunomodulatory compound administered orally to the gastrointestinal tract for the maintenance treatment and prevention of seasonal allergy symptoms (see, e.g., Hon et al., Drug Design, Development and Therapy, 8, 839 (2014)). It acts primarily by blocking the action of leukotriene D4 (as well as leukotrienes C4 and E4) on the cysteinal leukotriene receptor CysLT1 in the airway.

[0008] International Patent Applications WO 02 / 34237 and WO 2003 / 020200 disclose active agents covalently attached to polypeptides, more specifically to homo- and heteropolymers of naturally occurring and / or synthetic amino acids.

[0009] Unexpectedly, we previously discovered that montelukast exhibits anti-inflammatory effects when administered topically, for example, to the skin (see unpublished International Patent Application No. PCT / CN2018 / 094441). We have now discovered that covalently binding montelukast to specific amino acid sequences creates new compounds with potential use in a variety of conditions, including the topical treatment of skin conditions such as inflammation and wounds. Summary of the Invention

[0010] According to a first aspect of the present invention, there is provided a peptide-containing compound comprising a peptide component that is an amino acid sequence of 2 to 45 amino acids, wherein the peptide component is covalently attached to one or more compounds of formula I: [ka] During the ceremony, R 1 is -C(CH3)2OH, -COCH3, -C(CH3)=CH2, and —C(CH3)2H; a peptide-containing compound, wherein n is 0, 1, or 2; and positional isomers, stereoisomers, and pharmaceutically or cosmetically acceptable salts of the peptide-containing compounds, Hereinafter, the peptide-containing compounds, positional isomers, stereoisomers, and salts are collectively referred to as "compounds of the present invention."

[0011] The compounds of the present invention contain at least one primary amide linkage by which one or more compounds of Formula I are coupled to a peptide component. The amide linkage is formed by reaction of a carboxylic acid group(s) of one or more compounds of Formula I with one or more free -NH groups of the peptide component. The amide linkage can be formed at the N-terminal -NH group and / or through one or more other free -NH groups in amino acids present in the peptide sequence.

[0012] Therefore, it is preferred that at least one of the amino acids in the peptide component of the compound of the present invention contains a positively charged group (i.e., a free -NH group) that is not N-terminal. The amino acids in the peptide component of the compound of the present invention can be naturally occurring (but not necessarily proteogenic) amino acids and / or synthetic amino acids. Preferably, the amino acids in the peptide component are naturally occurring amino acids.

[0013] For example, at least one of the amino acids not present at the N-terminus of the peptide component may comprise asparagine (Asn), more preferably arginine (Arg), and even more preferably lysine (Lys).

[0014] In this regard, it is preferred that at least about 5%, more preferably at least about 10%, such as at least about 15%, including at least about 20%, of the amino acids present in the peptide component (by number and / or weight) comprise such amino acids (i.e., Asn, Arg, and / or Lys).

[0015] The peptide component of the compounds of the invention can be a homopolymer of amino acids. Alternatively, the peptide component of the compounds of the invention can be a heteropolymer of two or more different amino acids. Preferably, the peptide component of the compounds of the invention is a heteropolymer of two or more different naturally occurring amino acids (e.g., at least three different amino acids, preferably four different amino acids, and most preferably at least five different amino acids).

[0016] Peptide moieties that can be coupled to compounds of Formula I include peptides with known antibacterial and / or anti-inflammatory properties, or fragments or minor variants thereof.

[0017] Fragments of such peptides include "portions" of the complete amino acid sequence that may exhibit such antibacterial and / or anti-inflammatory properties.

[0018] Also included are amino acid sequences that are (e.g., minor) variants of such complete amino acid sequences or fragments thereof, which may be synthesized by chemical and / or biological processes (e.g., chemical modification of naturally occurring peptides or direct synthesis). By "(e.g., minor) variants of complete amino acid sequences or fragments thereof" is meant mutations in their respective sequences that do not measurably adversely affect the required properties of the relevant complete peptide or its fragment.

[0019] Antimicrobial peptides that may be mentioned in this regard include Cecropin A: H-Lys-Trp-Lys-Leu-Phe-Lys-Lys-Ile-Glu-Lys-Val-Gly-Gln-Asn-Ile-Arg-Asp-Gly-Ile-Ile-Ly s-Ala-Gly-Pro-Ala-Val-Ala-Val-Val-Gly-Gln-Ala-Thr-Gln-Ile-Ala-Lys-NH2 (SEQ ID NO: 1), Cecropin B: H-Lys-Trp-Lys-Val-Phe-Lys-Lys-Ile-Glu-Lys-Met-Gly-Arg-Asn-Ile-Arg-Asn-Gly-Ile-Val-Lys-Ala-Gly-Pro-Ala-Ile-Ala-Val-Leu-Gly-Glu-Ala-Lys-Ala-Leu-NH2 (SEQ ID NO: 2), and / or LL-37: Cecropins such as H-Leu-Leu-Gly-Asp-Phe-Phe-Arg-Lys-Ser-Lys-Glu-Lys-Ile-Gly-Lys-Glu-Phe-Lys-Arg-Ile-Val-Gln-Arg-Ile-Lys-Asp-Phe-Leu-Arg-Asn-Leu-Val-Pro-Arg-Thr-Glu-Ser-NH2 (SEQ ID NO: 3) are included.

[0020] Anti-inflammatory peptides that may be mentioned include 11 identified individual adhesive protein subtypes derived from mussels, including collagens pre-COL-P, pre-COL-D, and pre-COL-NG; mussel adhesive proteins (MAPs), also known as mussel foot proteins (mefps), which are proteins secreted by marine shellfish species such as Mytilus edulis, Mytilus coruscus, and Perna viridis; mussel foot matrix proteins PTMP (proximal thread matrix protein) and DTMP (distal-proximal thread matrix protein); and mfp proteins mfp-2 (sometimes referred to as "mefp-2" and hereafter used interchangeably), mfp-3 / mefp-3, mfp-4 / mefp-4, mfp-5 / mefp-5, mfp-6 / mefp-6, and most preferably mfp-1 / mefp-1 (see, e.g., Zhu et al., Advances in Marine Science, 32, 560 (2014) and Gao et al, Journal of Anhui Agr. Sci., 39, 19860 (2011)).

[0021] The majority of mefp-1 consists of 70-90 tandem repeats of the decapeptide Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (SEQ ID NO: 4; see Waite, Int. J. Adhesion and Adhesives, 7, 9 (1987)). This decapeptide sequence can be isolated as a low-molecular-weight derivative of naturally occurring MAP or synthesized as described, for example, by Yamamoto in J. Chem. Soc., Perkin Trans. 1, 613 (1987). See also Dalsin et al., J. Am. Chem. Soc., 125, 4253 (2003).

[0022] In this regard, peptide components that may be used in the compounds of the present invention comprise one to four (preferably three, more preferably two) repeating units of the above-mentioned decapeptide sequences, or most preferably one of the above-mentioned decapeptide sequences, or a variant (as defined above) of any of these options.

[0023] Further provided is a peptide-containing compound as defined above, wherein at least about 5%, e.g., at least about 10% (by number) of the amino acids in the peptide component contain an aromatic group such as tyrosine and / or 3,4-dihydroxyphenylalanine (DOPA). Alternatively, none of the amino acids in the peptide chain contain an aromatic group, i.e., none of the amino acids are tyrosine and / or DOPA.

[0024] More preferably, the peptide component in the compound of the invention comprises 5 to 30 amino acids, such as 6 to 20 amino acids, including 7 to 15 (e.g., 10, up to 12) amino acids in the amino acid sequence.

[0025] Compounds of the invention which may be mentioned in this regard include those in which the peptide component has the amino acid sequence: X-Pro-YZ (SEQ ID NO: 5), During the ceremony, X represents a chain of 1 to 2 amino acid residues independently selected from the group consisting of Ala and Lys; Y is selected from the group consisting of Ser and pSer; Z represents a chain of 1 (e.g., 2, such as 3) to 7 amino acid residues each independently selected from the group consisting of Tyr, pTyr, Hyp (i.e., 3Hyp or 4Hyp), Thr, pThr, DOPA, and Lys; At least one of the Ala and / or Lys residues is attached to one or more compounds of formula I as defined above.

[0026] A particular amino acid sequence that may be mentioned is Ala-Pro-Ser-Hyp-Hyp-Thr (SEQ ID NO: 6).

[0027] Preferred compounds of the invention that may be mentioned include those in which the peptide component has the amino acid sequence: G 1 -Lys-Pro-G 2 -T-Hyp-G 3 -Lys (SEQ ID NO: 7) wherein: G 1 represents absence or Ala, G 2 is selected from the group consisting of Ser and pSer; T is selected from the group consisting of DOPA, or more preferably, Tyr and pTyr; Hyp is selected from the group consisting of 3Hyp and 4Hyp; G 3 represents a chain of 1 to 4 (e.g., 1 to 3) amino acid residues independently selected from the group consisting of Tyr, pTyr, 3Hyp, 4Hyp, Thr, pThr, and DOPA; At least one of the Lys residues and / or Ala residues, if present, is attached to one or more compounds of formula I as defined above.

[0028] Peptide components that may be mentioned include those with the amino acid sequence: Lys-Pro-G 2 -T-Hyp-G 3 -Lys (SEQ ID NO: 8) wherein G 2 , T, Hyp, and G 3 is defined above, and G 2 is more preferably Ser, and G 3 is more preferably Tyr or especially DOPA.

[0029] However, it is more preferable that: G 1 represents Ala, G 2 represents Ser, T represents Tyr, G3 -V 1 -Thr-Tyr-V 2 and V represents 1 to 4 (e.g., 1 to 3) amino acid residues independently selected from the group consisting of Tyr, 3Hyp, 4Hyp, and Thr, such as a chain of amino acids represented by 1 binds to Hyp and V 2 is attached to Lys and V 1 and V 2 independently represent none, one or two Hyp groups.

[0030] In this regard, Array-V 1 -Thr-Tyr-V 2 is a subarray: -Hyp-Thr-Tyr- -Hyp-Thr-Tyr-Hyp-, or The order of priority can be expressed as -Thr-Tyr-Hyp-.

[0031] Thus, preferred peptide components include the amino acid sequence: Ala-Lys-Pro-G 2 -T-Hyp-Hyp-Thr-G 4 -Lys (SEQ ID NO: 9) wherein G 2 , T, and Hyp are defined above, and G 4 is selected from the group consisting of Tyr, pTyr, 3Hyp, 4Hyp, Thr, pThr, and DOPA; 2 is more preferably Ser, T is more preferably Tyr, and G 4 is more preferably Tyr or DOPA.

[0032] A preferred peptide component has the amino acid sequence: Ala-Lys-Pro-G 2 -T-Hyp-Thr-G 4 -Hyp-Lys (SEQ ID NO: 10) Further included are those in which: 2 , T, Hyp, and G 4is defined above, and G 4 is more preferably DOPA, or especially Tyr.

[0033] A preferred peptide component has the amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (SEQ ID NO: 4), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-DOPA-Lys (SEQ ID NO: 11), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 12), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 13), Lys-Pro-Ser-Tyr-Hyp-DOPA-Lys (SEQ ID NO: 14), and Lys-Pro-Ser-pTyr-Hyp-DOPA-Lys (SEQ ID NO: 15) is included.

[0034] Of the above list of amino acid sequences, Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 12) is the most preferred.

[0035] Further preferred peptide components include the amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-DOPA-Lys (SEQ ID NO: 16) and The amino acid sequence of the ...

[0036] Further preferred peptide components include the amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 18), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 19), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 20), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 21), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 22), Ala-Lys-Pro-pSer-Tyr-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 23), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 24), Ala-Lys-Pro-pSer-Tyr-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 25), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-DOPA-Hyp-Lys (SEQ ID NO: 26), and This includes Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-Tyr-Hyp-Lys (SEQ ID NO: 27).

[0037] Of the above list of amino acid sequences, Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 24) is most preferred.

[0038] Further preferred peptide components include the amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA (SEQ ID NO: 28), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-DOPA (SEQ ID NO: 29), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr (SEQ ID NO: 30), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr (SEQ ID NO: 31), Lys-Pro-Ser-Tyr-Hyp-DOPA (SEQ ID NO: 32), Lys-Pro-Ser-pTyr-Hyp-DOPA (SEQ ID NO: 33), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-DOPA (SEQ ID NO: 34), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-Tyr (SEQ ID NO: 35), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp (SEQ ID NO: 36), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-DOPA-Hyp (SEQ ID NO: 37), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp (SEQ ID NO: 38), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr-Hyp (SEQ ID NO: 39), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp (SEQ ID NO: 40), Ala-Lys-Pro-pSer-Tyr-Hyp-Thr-DOPA-Hyp (SEQ ID NO: 41), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp (SEQ ID NO: 42), Ala-Lys-Pro-pSer-Tyr-Hyp-Thr-Tyr-Hyp (SEQ ID NO: 43), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-DOPA-Hyp (SEQ ID NO: 44), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-pThr-Tyr-Hyp (SEQ ID NO: 45), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp (SEQ ID NO: 46), and Ala-Lys-Pro-Ser-Tyr-Hyp (SEQ ID NO: 47) is included.

[0039] Those skilled in the art will appreciate that metabolic products of the compounds of the invention that may be formed following administration are included within the scope of the present invention.

[0040] In particular, compounds of the present invention that contain peptide components having the amino acid sequences of SEQ ID NO:28 through SEQ ID NO:47 may be formed as metabolites of the corresponding compounds of the present invention that contain amino acids associated with the C-terminus that may be cleaved from other compounds of the present invention following administration.

[0041] For example, compounds of the invention that include peptide components having the amino acid sequences of SEQ ID NO:30, SEQ ID NO:46, and SEQ ID NO:47 may be formed as metabolites of compounds of the invention that include peptide components having the amino acid sequence of SEQ ID NO:12. Similarly, compounds of the invention that include peptide components having the amino acid sequences of SEQ ID NO:42 and SEQ ID NO:47 may be formed as metabolites of compounds of the invention that include peptide components having the amino acid sequence of SEQ ID NO:24.

[0042] Nevertheless, compounds of the invention that include peptide components having the amino acid sequences of SEQ ID NO:28 through SEQ ID NO:47 are also themselves compounds of the invention and can be made, formulated, and administered to patients in exactly the same manner as other compounds of the invention described herein and / or exemplified below.

[0043] For the avoidance of doubt, as used herein, Pro represents proline, Ala represents alanine, Ser represents serine, pSer represents phosphoserine, Tyr represents tyrosine, pTyr represents phosphotyrosine, and Hyp represents hydroxyproline (3Hyp represents 3-hydroxyproline, and 4Hyp represents 4-hydroxyproline). Thr represents threonine, pThr represents phosphothreonine (or phosphonothreonine), and Lys, Ala, and DOPA are as defined above. Phosphonated derivatives of amino acids containing a free hydroxy group (e.g., Ser, Tyr, and Thr) contain an -OP(=O)(OH) group instead of -OH.

[0044] One or more compounds of Formula I can be linked to the aforementioned peptide components via one or more, for example, Ala or Lys moieties (including an N-terminal Ala or Lys moiety and / or a C-terminal Lys moiety).

[0045] Three compounds of formula I, preferably two compounds of formula I, more preferably one compound of formula I, can be covalently attached to the aforementioned peptide moiety in the compounds of the invention.

[0046] Advantageously, the peptide component of the compounds of the present invention comprises at least one amino acid residue containing at least one free -NH2 group. Particular amino acid residues containing at least one free -NH2 group include Asn, Gln, preferably Arg, and most preferably Lys. For example, the peptide component may comprise G 1 -Lys-Pro-G 2 -T-Hyp-G 3 -Lys (SEQ ID NO: 7), at least one of the lysine residues has a free -NH2 group (i.e., at least one of the Lys residues is not covalently attached to a compound of formula I).

[0047] In examples of compounds of the invention in which two or more (e.g., three, preferably two) compounds of Formula I are covalently attached to a peptide component, the peptide component preferably comprises at least one amino acid residue containing at least one free -NH group. For example, the peptide component comprises at least one Lys residue that is not covalently attached to a compound of Formula I.

[0048] Without wishing to be bound by theory, it is believed that the presence of a free -NH group in the peptide component (e.g., when one or more of the possible Lys residues is not covalently attached to the compound of Formula I) is beneficial to the (e.g., aqueous) solubility of the compounds of the invention.

[0049] Compounds of formula I contain a double bond and may therefore exist as E (entgegen) and Z (zusammen) geometric isomers about the double bond. All such isomers and mixtures thereof are included within the scope of the present invention. For the avoidance of doubt, [ka] indicates that in the compound, the 7-chloroquinoline ring can be positioned either cis (as a Z geometric isomer) or trans (as an E geometric isomer) across the double bond to the central 1,3-disubstituted phenyl ring. Preferably, the 7-chloroquinoline ring is positioned trans across the double bond to the central 1,3-disubstituted phenyl ring, i.e., the E geometric isomer.

[0050] Particular compounds of formula I that may be mentioned include those in which R 1 is selected from the group consisting of -C(CH3)2OH, -COCH3, and -C(CH3)=CH2.

[0051] Preferred compounds of formula I include R 1 is —C(CH 3 ) 2 OH and / or n is 0 (e.g., montelukast).

[0052] For the avoidance of doubt, montelukast has the following chemical structure: [ka]

[0053] Other compounds of formula I include R 1 is -C(CH3)=CH2 and / or n is 0 (e.g., montelukast styrene).

[0054] For the avoidance of doubt, montelukast styrene has the following chemical structure: [ka]

[0055] Other compounds of formula I that may be mentioned include those in which R 1 but Included are those where n is -C(CH3)2H and / or n is 0 (e.g., hydrogenated montelukast styrene).

[0056] For the avoidance of doubt, hydrogenated montelukast styrene has the following chemical structure: [ka]

[0057] The compounds of the present invention, whether in the form of a salt or not, include positional isomers (e.g., Hyp and Tyr moieties) within the amino acids of the peptide components, as well as mixtures of such positional isomers. For example, the definition of Tyr includes not only tyrosine (4-hydroxyphenylalanine) but also 2- and 3-hydroxyphenylalanine, and the definition of Hyp includes 4-hydroxyproline (4Hyp), 3-hydroxyproline (3Hyp), and 5-hydroxyproline (5Hyp). It is more preferred that the Hyp residue is 4-hydroxyproline.

[0058] Also, in addition to the standard central carbon atom of the amino acids in the peptide component of the compounds of the present invention (usually, but not exclusively, in the L-configuration), certain amino acids within the sequence contain an additional chiral carbon atom. All such stereoisomers and mixtures thereof (including racemic mixtures) are included within the scope of the present invention. In comparison, the definition of Hyp includes trans-4-hydroxy-L-proline, cis-4-hydroxy-L-proline, trans-3-hydroxy-L-proline, cis-3-hydroxy-L-proline, trans-5-hydroxy-L-proline, and cis-5-hydroxy-L-proline, although Hyp used in the compounds of the present invention is preferably 4-hydroxy-L-proline. Similarly, individual enantiomers of compounds of Formula I that may form part of the compounds of the present invention are included within the scope of the present invention.

[0059] The compound of the present invention may be in the form of salt.The salts that can be mentioned include pharmaceutically acceptable salts and / or cosmetically acceptable salts, such as pharmaceutically and / or cosmetically acceptable acid addition salts and base addition salts.Such salts can be formed by conventional means, for example, by reacting the compound of the present invention with one or more equivalents of a suitable acid or base in any solvent or in a medium in which the salt is insoluble, and then removing the solvent or the medium using standard techniques (for example, by vacuum, lyophilization or filtration).Salts can also be prepared by exchanging the counterion of the active ingredient in the form of salt with another counterion, for example, by using a suitable ion exchange resin.

[0060] Preferred salts include, for example, acetate, hydrochloride, bisulfate, maleate, mesylate, tosylate, alkaline earth metal salts such as calcium and magnesium, or alkali metal salts such as sodium and potassium salts. Most preferably, the compounds of the present invention may be in the form of acetate salts.

[0061] The compounds of the present invention can be prepared by conventional techniques. For example, the compounds of the present invention can be made by coupling one or more compounds of formula I to a peptide component, for example as described below. The compounds of the present invention (and their peptide components) can be synthesized from available starting materials using appropriate reagents and reaction conditions. In this regard, those skilled in the art can refer in particular to "Comprehensive Organic Synthesis" by BM Trost and I. Fleming, Pergamon Press, 1991. Further references that can be used include "Heterocyclic Chemistry" by JA Joule, K. Mills and GF Smith, 3 rdedition, published by Chapman & Hall, "Comprehensive Heterocyclic Chemistry II" by A.R. Katrittzky, C.W. Rees and E.F.V. Scriven, Pergamon Press, 1996, and "Science of Synthesis", Volumes 9-17 (Hetarenes and Related Ring Systems), Georg Thieme Verlag, 2006.

[0062] Peptide components of the components of the invention may be made, if desired, by standard peptide synthesis techniques using standard amino acid coupling techniques, and standard coupling reagents and solvents, for example as described below.

[0063] Those skilled in the art will understand that the substituents defined herein and thereon may be modified one or more times after or during the processes described above for preparing the compounds of the present invention by methods well known to those skilled in the art. Examples of such methods include substitution, reduction, oxidation, dehydrogenation, alkylation, dealkylation, acylation, hydrolysis, esterification, etherification, halogenation, and nitration. Precursor groups can be converted to different such groups or to groups defined in the compounds of the present invention at any time during the reaction sequence. Those skilled in the art may also refer to "Comprehensive Organic Functional Group Transformations" by A.R. Katrittzky, O. Meth-Cohn and C.W. Rees, Pergamon Press, 1995, and / or "Comprehensive Organic Transformations" by R.C. Larock, Wiley-VCH, 1999.

[0064] The compounds of the invention can be isolated from their reaction mixtures and, if necessary, purified using conventional techniques such as those known to those skilled in the art. Thus, the processes for preparing the compounds of the invention described herein may comprise, as a final step, the isolation and optional purification of the compounds of the invention.

[0065] It will be appreciated by those skilled in the art that in the processes described above and below, the functional groups of intermediate compounds may need to be protected by protecting groups. Protection and deprotection of functional groups may be carried out before or after the reactions in the schemes mentioned above.

[0066] Protecting groups can be applied and removed according to techniques well known to those skilled in the art and as described below. For example, protected compounds / intermediates described herein can be chemically converted to unprotected compounds using standard deprotection techniques. The type of chemistry involved will dictate the need and type of protecting groups and the sequence for achieving synthesis. The use of protecting groups is fully described in "Protective Groups in Organic Synthesis", 3rd edition, T.W. Greene & P. ​​G.M. Wutz, Wiley-Interscience (1999), the contents of which are incorporated herein by reference.

[0067] The compounds of the present invention are useful because they have pharmacological activity.Therefore, the compounds of the present invention are useful as human and animal medicines.Therefore, they are indicated as pharmaceuticals (and / or veterinary medicine), but can also be used as cosmetics and / or as part of medical devices.

[0068] While the compounds of the invention may have pharmacological activity themselves, certain pharmaceutically acceptable (e.g., "protected") derivatives of the compounds of the invention exist or may be prepared, which may have no such activity but can be orally administered and subsequently metabolized or chemically converted in the body to form the compounds of the invention. Such compounds (which may have some pharmacological activity, provided that such activity is appreciably less than the activity of the active compound to which they are metabolized / converted) may therefore be described as "prodrugs" of the compounds of the invention.

[0069] As used herein, reference to a prodrug will include a compound that forms an experimentally detectable amount of a compound of the invention within a given time period after administration. All prodrugs of the compounds of the invention are included within the scope of the invention.

[0070] The compounds of the present invention are particularly useful in the treatment of inflammation.

[0071] "Treatment of inflammation" includes treatment of inflammation in any organ of the body (including soft tissues, joints, nerves, vasculature, internal organs, particularly mucosal surfaces, and specifically skin), regardless of cause, and also includes all such inflammatory disorders or conditions and / or disorders or conditions characterized (e.g., as a symptom) by inflammation.

[0072] Inflammatory conditions can be (and typically are) characterized by the activation of immune defense mechanisms, resulting in effects that are more harmful than beneficial to the host. Such conditions are generally associated with varying degrees of tissue redness or hyperemia, swelling, edema, hyperthermia, pain (including soreness), fluid exudation, itching (pruritus), cell death, and tissue destruction, cell proliferation, and / or loss of function.

[0073] Inflammatory conditions that may be mentioned include arteritis, diabetes, metabolic syndrome, rosacea, asthma and allergy, ankylosing spondylitis, chronic obstructive pulmonary disease, gouty arthritis, inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), multiple sclerosis, osteoarthritis, pancreatitis, prostatitis, psoriatic arthritis, rheumatoid arthritis, tendonitis, bursitis, Sjogren's syndrome, systemic lupus erythematosus, uveitis, urticaria, vasculitis, mastocytosis, diabetic vascular complications, migraine, atherosclerosis and related cardiovascular disorders.Another condition characterized by inflammation that may be mentioned is chronic obstructive pulmonary disease (COPD).Another condition characterized by inflammation that may be mentioned is inflammatory bowel disease, including Crohn's disease and especially ulcerative colitis.

[0074] Further inflammatory conditions that may be mentioned in particular include inflammation of the skin or mucous membranes (including the oral, nasal, ocular, vaginal, cervical, and / or anorectal mucosa, more particularly the oral or nasal mucosa), such as inflammation resulting from infection (such as viral and / or bacterial infection) or allergic / atopic conditions (such as rhinitis (e.g., allergic rhinitis), pharyngitis, periodontitis, gingivitis, xerophthalmia, conjunctivitis (e.g., allergic conjunctivitis), dermatitis, urticaria (hives), and food allergies), as well as other inflammatory conditions such as herpes, drug eruptions, polymorphous light eruption, sunburn, early symptoms of skin cancer (erythematous skin lesions), pathological alopecia (including after skin grafts), chemical rash, psoriasis, erythema multiforme, folliculitis, eczema, and otitis externa. A disease state that may be mentioned is polymorphous light eruption.

[0075] More specifically, compound can be used for treating certain pathologies characterized by inflammation and / or associated with inflammation.Such pathologies can include wounds (including abrasions (scratches), incisions (including surgical incisions), lacerations, punctures, avulsions, contusions and scars), burns (including inflammation caused by post-burn surgery, such as skin grafting), and other pathologies such as hemorrhoids.Wounds can be acute or chronic, and / or can be caused by one or more inflammatory disorders as defined herein.

[0076] Skin or mucosal wounds can result from internal or external physical injury to the membrane surface, or can be caused by (ie, be a symptom of) an underlying physiological disorder.

[0077] Physical (e.g., "open") wounds can be caused by sharp objects (cuts, incisions, punctures) or blunt objects / mechanical forces (lacerations, abrasions, avulsions), physical blows (contusions), heat or chemicals (burns and blisters), ultraviolet light (sunburn), cold (chills or frostbite). Wounds can be superficial (damage to the epidermis and / or dermis only) or full-thickness wounds (damage below the epidermis and / or dermis). In severe cases, subcutaneous and / or submucosal tissues, such as muscles, bones, joints, and even internal organs, may be damaged.

[0078] The compounds of the present invention can be used to alleviate pain (including pain) associated with inflammation and / or wounds. Specifically, the compounds of the present invention can be used to alleviate procedural pain and / or non-procedural pain. Those skilled in the art will understand that the term "procedural pain" (i.e., surgical pain) refers to acute pain associated with medical investigation and treatment carried out for medical purposes. The term "non-procedural" refers to general pain associated with inflammation and / or wounds (e.g., pain associated with dental ulcers, burns, and / or scars), and is not the result of a specific medical intervention.

[0079] The compounds of the invention can be used to treat inflammation, pain (including pain), and / or pruritus (itching) associated with the wound itself and the healing process, as well as to prevent exudation of fluid from the wound, the risk of infection, and physiological responses resulting from inflammation and / or the wound healing process, such as scarring and melanin pigmentation.

[0080] Scarring is the result of inflammation and / or wound healing and is a general term for the formation of fibrous tissue that is the result of such inflammation / healing.

[0081] The compound of the present invention can also be useful for suppressing the production of melanin pigmentation, which may or may not be caused by inflammation and / or wound healing.The compound of the present invention can also be useful for suppressing the pigmentation caused by skin diseases such as melanoma, freckles, melanin pigmentation, butterfly rash and other pigmentation-related disorders, and melanoma-related skin cancer, as well as sun exposure or acne.

[0082] Wounds can also occur as a result of disease or injury (e.g., inflammation). Such wounds can include blisters and / or ulcers of the skin and mucous membranes. These are common conditions that are often long-lasting and difficult to treat. Skin tissue can often be damaged, removed, liquefied, infected, and / or necrotic. Ulcers, especially if infected, can have secondary health consequences and are difficult and expensive to heal. They can also cause significant psychological stress and economic loss to patients, affecting both their overall well-being and quality of life.

[0083] Alternatively, inflammatory skin conditions or diseases in which the compounds of the invention find particular utility include the treatment of psoriasis, acne, eczema and dermatitis, particularly allergic / atopic dermatitis, as well as rhinitis, particularly allergic rhinitis, haemorrhoids, chronic obstructive pulmonary disease, and mucositis characterized by ulcerative colitis.

[0084] Psoriasis is a chronic inflammatory skin disease that tends to recur (some patients never recover). Clinical symptoms of psoriasis mainly include erythema and scaling. It can occur all over the body, but is more commonly observed on the scalp and hands and feet.

[0085] Acne is a chronic inflammatory skin disease of the follicular (pilosebaceous) system, the development of which is closely related to factors such as excessive sebum, obstruction of the pilosebaceous ducts (including closed and open comedones), bacterial infection, and inflammatory reactions. It tends to occur at a young age and is characterized by polymorphic skin lesions on the face. Therefore, the term acne includes both ordinary acne and acne rosacea (i.e., red nose).

[0086] Eczema is an intensely itchy inflammatory reaction of the skin caused by various internal and external factors. It has three stages: acute, subacute, and chronic. The acute stage tends to produce exudate, while the chronic stage involves infiltration and hypertrophy. Skin lesions are often itchy and easily recur.

[0087] Dermatitis is a common skin disorder characterized by roughness, redness, itchiness, eczema, and dryness. The small lumps, intractable ulcers, and pigmented patches caused by dermatitis can develop into basal cell carcinoma, squamous cell carcinoma, and malignant melanoma if not treated promptly. Dermatitis can be caused by a variety of internal and external infectious or non-infectious factors, including substances (contact dermatitis) or allergies (allergic / atopic dermatitis). Also included are seborrheic dermatitis (seborrheic eczema) and all forms of steroid-dependent dermatitis (including photosensitive seborrheic dermatitis, perioral dermatitis, rosacea-like dermatitis, steroid rosacea, steroid-induced rosacea, iatrosacea, steroid-dermatitis-like rosacea, topical corticosteroid-induced rosacea-like dermatitis, and more specifically, facial corticosteroid-addicted dermatitis (FCAD) or facial corticosteroid-dependent dermatitis (FCDD), characterized by flushing, erythema, telangiectasias, atrophy, papules, and / or pustules in the facial area after long-term treatment (including uncontrolled use, abuse, or misuse) with topical corticosteroids (see, e.g., Xiao et al., J. Dermatol., 42, 697 (2015) and Lu et al., Clin. & Exp. Dermatol., 35, 618 (2009)).

[0088] Rhinitis is the irritation and inflammation of the mucous membrane inside the nose.The common symptoms of rhinitis include stuffy nose, runny nose, sneezing and postnasal drip.The most common type of rhinitis is allergic rhinitis, which is caused by allergens such as pollen, dust, mold or certain animal skin flakes.Surprisingly, it has been found that the patients with allergic rhinitis treated with the compound of the present invention can experience relief of itchy eyes, even when the compound of the present invention is administered intranasally (i.e., to the nasal mucous membrane).

[0089] Hemorrhoids are swellings caused by large inflammations of the hemorrhoidal blood vessels found in or around the rectum and anus. Symptoms include bleeding (i.e., wounds) after passing stool, prolapsed hemorrhoids, mucus secretion, and itching, burning, redness, and swelling in the anal area. Hemorrhoids are thought to be the result of increased abdominal pressure, for example, as a result of constipation or diarrhea.

[0090] Chronic obstructive pulmonary disease (COPD) is the name for a group of lung conditions that cause difficulty breathing, including emphysema (damage to the alveoli) and chronic bronchitis (long-term airway inflammation). COPD occurs when the lungs become inflamed, damaged, and narrowed. The damage to the lungs is usually irreversible, resulting in impaired airflow in and out of the lungs. Symptoms of COPD include shortness of breath, a productive cough, frequent chest infections, and persistent wheezing. The most common cause of the disease is smoking, but other risk factors include high levels of air pollution and occupational exposure to dust, chemicals, and smoke.

[0091] The compounds of the present invention may have a positive effect on the relief of erythema, redness and swelling, edema, blisters, and bullous pemphigoid caused by various conditions, including those generally and specifically mentioned herein, and may inhibit the exudation of subcutaneous tissue fluid and suppress the itching and pain caused by such inflammatory conditions.

[0092] Other inflammatory conditions that may be mentioned include: (a) Mucosal inflammation such as inflammation caused by oral mucositis, aphthous ulcers, otitis media, laryngitis, tracheitis, esophagitis, gastritis, enteritis, and enterocolitis (including bacterial dysentery, chronic amebic dysentery, schistosomiasis, nonspecific ulcerative colitis, and regional enteritis), cervicitis and endocervicitis, endometritis, inhalation injuries, and the like, as well as inflammation of the mucosa associated with cancer and infections (e.g., viral infections such as the common cold or influenza) affecting mucosal surfaces such as the oral cavity, nasopharynx, ears, throat, trachea, gastrointestinal tract, and cervix. (b) For example, fractures, pyogenic infections of bones and joints, inflammation due to rheumatic bone disease, and pyogenic osteomyelitis (acute, chronic, localized, sclerotic, post-traumatic), septic arthritis, bone tumors (osteoma, osteoid osteoma, chondroma), bone cysts, osteoclastoma, primary bone sarcomas (osteosarcoma, chondrosarcoma, osteofibrosarcoma, Ewing's sarcoma, non-Hodgkin's lymphoma, myeloma, chordoma), metastatic bone tumors, tumor-like lesions of bone (bone cysts, aneurysmal bone cysts, eosinophilic granuloma, fibrous dysplasia), and orthopedic inflammation associated with rheumatoid arthritis. (c) Nerve inflammation such as peripheral polyneuritis, facial neuritis, peripheral neuritis, subcutaneous neuritis, ulnar neuritis, and intercostal neuritis. (d) Inflammation of subcutaneous and submucosal soft tissues, such as myositis, dendritis, tendonitis, cystitis, lymphadenitis, cryptitis, tonsillitis, synovitis, and fasciitis, as well as inflammation of soft tissues caused by injury, contusion, or tear of muscle, ligament, fascia, tendon, synovium, fat, joint capsule, and lymphatic tissue. (e) Vascular inflammation such as allergic leukocytoclastic vasculitis, allergic cutaneous vasculitis, polyarteritis nodosa, thromboangiitis, granulomatous vasculitis, lymphocytic vasculitis, vasculitis with abnormal blood composition, and rheumatoid vasculitis, as well as vascular inflammation associated with vascular cancer caused by allergic leukocytoclastic vasculitis, polyarteritis nodosa, thromboangiitis, granulomatous vasculitis, lymphocytic vasculitis, vasculitis with abnormal blood composition, and rheumatoid vasculitis. (f) Inflammation of internal organs such as the heart, stomach, intestines, lungs, liver, spleen, kidneys, pancreas, bladder, ovaries, and prostate, including, but not limited to, the treatment of pericarditis, myocarditis, endocarditis, pneumonia, hepatitis, splenitis, nephritis, pancreatitis, cystitis, oophoritis, prostatitis, and gastric ulcers. (g) Inflammation of the eye and surrounding area, such as conjunctivitis, keratitis (e.g., acute superficial keratitis, nummular keratitis, interstitial keratitis, disciform keratitis, neurotrophic keratitis, mucoplaque keratitis, herpes simplex keratitis, herpes zoster keratitis, bacterial keratitis, fungal keratitis, acanthamoeba keratitis, Onchocerca volvulus keratitis, superficial punctate keratitis, ulcerative keratitis, lagophthalmos keratitis, photokeratitis, and acute hyperemia during contact lens wear), and optic neuritis. (h) Inflammation of the gums and oral cavity, such as periodontitis, gingivitis, and dental ulcers. (i) Rheumatoid inflammation such as rheumatoid vasculitis, rheumatoid arthritis, rheumatic bone disease, ankylosing spondylitis, bursitis, Crohn's disease, gout, infectious arthritis, juvenile idiopathic arthritis, osteoarthritis, osteoporosis, polymyalgia rheumatica, polymyositis, psoriatic arthritis, scleroderma, Sjogren's syndrome, spondyloarthropathy, systemic lupus erythematosus, and tendonitis.

[0093] The compounds of the present invention can also be used to treat certain special diseases of the respiratory system, such as pulmonary cystic fibrosis, common interstitial pneumonia, allergic pneumonia, asbestosis, emphysema, cor pulmonale, pulmonary embolism, etc. A special disease state that may be mentioned is idiopathic pulmonary fibrosis.

[0094] Idiopathic pulmonary fibrosis is a diffuse and fatal pulmonary interstitial disease, with pathological characteristics including alveolar epithelial damage, extensive proliferation of pulmonary fibroblasts, and excessive deposition of extracellular matrix, which ultimately leads to irreversible lung tissue damage.In the later stage of the disease, the subject with idiopathic pulmonary fibrosis experiences respiratory failure and death.It has been found that the compound of the present invention can be useful in treating idiopathic pulmonary fibrosis and / or alleviating the symptoms associated with this disease.

[0095] The compounds of the invention are useful in treating the following pulmonary and / or fibrotic conditions (whether or not otherwise described herein): pulmonary fibrosis, renal fibrosis, hepatic fibrosis, silicosis, acute bronchitis, chronic bronchitis, tracheobronchitis, bronchial asthma, severe asthma, bronchiectasis, upper respiratory tract infections including colds and influenza, allergic airway inflammation, bacterial pneumonia, viral pneumonia, mycoplasmal pneumonia, reckettsia, radiation pneumonitis, pneumococcal (including staphylococcus, streptococcus, and gram-negative bacillus) pneumonia, pulmonary candidiasis (including aspergillosis, mucormycosis, histoplasmosis, actinomycosis, and nocardiosis), pulmonary mycoses, cryptococcosis, lung abscess, anaphylactic pneumonia (Leoffer's syndrome), and / or pulmonary fibrosis. It is particularly useful in the treatment of pulmonary eosinophilia (eosinophilic syndrome), extrinsic allergic alveolitis, pulmonary eosinophilia (hypereosinophilia), obstructive emphysema, pulmonary edema, pulmonary tuberculosis, respiratory alkalosis (acidosis), acute lung injury, interstitial lung disease, empyema, pulmonary fibroma, and cor pulmonale.

[0096] Particular mucosal disorders and diseases in which the compounds of the invention are found to be useful include anorectal diseases such as diarrhea, hemorrhoids, abscesses, fistulas, anal fissures, pruritus ani, anal sinusitis, warts and rectal prolapse; inflammatory bowel diseases including Crohn's disease, especially ulcerative colitis; gynecological diseases such as cervicitis, vaginitis, pelvic pain and disorders; and dental diseases such as paradentitis.

[0097] The compounds of the present invention may further have antioxidant effects by increasing SOD (superoxide dismutase) production and reducing lipid oxidation, and therefore the compounds of the present invention may be considered to have antioxidant properties.

[0098] The compounds of the present invention may also have antipyretic properties, allowing for the treatment of fever and / or alleviating its symptoms, for example, by reducing the body temperature of a subject (which results in a reduction in fever). Thus, the compounds of the present invention and formulations containing them may be considered to be antipyretics.

[0099] According to a further aspect of the present invention, there is provided a method for treating inflammation, inflammatory disorders, and / or disorders / conditions characterized by inflammation (e.g., as a symptom), comprising administering a compound of the present invention to a patient in need of such treatment.

[0100] For the avoidance of doubt, in the context of the present invention, the terms "treatment", "therapy" and "treatment" include therapeutic or palliative treatment of patients in need of treatment, as well as prophylactic treatment and / or diagnosis of patients susceptible to inflammation and / or inflammatory disorders.

[0101] The compounds of the present invention may further have antiviral properties that may allow for the treatment of viral infections or viral diseases by interfering with viral replication within the host, as opposed to treating any viral infection or any symptoms of a viral infection, such as pain and / or inflammation. Such antiviral properties may also allow for the prevention of the onset of such infections or diseases, the protection of cells within the host from (e.g., further) viral infections, the prevention or arrest of the spread of viral infections or diseases (within a single host or from one host to new hosts), or the prevention of viral reactivation after latency in the host.

[0102] According to a further aspect of the present invention there is provided a method for treating a viral infection which comprises administering to a patient in need of such treatment a compound of the present invention, or a salt thereof.

[0103] Viral infections that may be mentioned include adenoviridae (e.g., adenovirus), papillomaviridae (e.g., human papillomavirus), polyomaviridae (e.g., BK virus, JC virus), herpesviridae (e.g., herpes simplex type 1, herpes simplex type 2, varicella-zoster virus, Epstein-Barr virus, human cytomegalovirus, human herpesvirus type 8), poxviridae (e.g., smallpox), hepadnaviridae (e.g., Hepatitis B virus), parvoviridae (e.g., parvovirus B19), astroviridae (e.g., human astrovirus), caliciviridae (e.g., norovirus, Norwalk virus), picornaviridae (e.g., coxsackievirus, hepatitis A virus, poliovirus, rhinovirus), coronaviridae (e.g., acute respiratory syndrome virus), flaviviridae (e.g., hepatitis C virus, yellow fever virus, dengue virus, West Nile virus), virus, tick-borne encephalitis virus), retroviridae (e.g., human immunodeficiency virus, HIV), togaviridae (e.g., rubella virus), arenaviridae (e.g., Lassa virus), bunyaviridae (e.g., hantavirus, Crimean-Congo hemorrhagic fever virus, hantavirus), filoviridae (e.g., Ebola virus, Marburg virus, Ravn virus), orthomyxoviridae (e.g., influenza A viruses (e.g., H1N1 and H3N2 viruses) Viruses not assigned to a family, such as influenza viruses, influenza B viruses, or influenza C viruses, paramyxoviridae (e.g., measles virus, mumps virus, parainfluenza virus, respiratory syncytial virus), rhabdoviridae (e.g., rabies virus), hepeviridae (e.g., hepatitis E virus), reoviridae (e.g., rotavirus, orbivirus, coltivirus, bannavirus), and hepatitis D virus, are also included.

[0104] Viruses that may be mentioned more specifically include herpes simplex viruses type 1 and type 2, human papilloma viruses, influenza viruses, and parainfluenza viruses.

[0105] The compounds of the present invention may further have antibacterial and / or bacteriostatic properties that may allow for the treatment of the bacterial infection itself, i.e., the treatment of a bacterial infection or disease by interfering with the growth or proliferation of bacteria within the host, as opposed to treating any symptoms of the bacterial infection or disease, such as pain and / or inflammation. Thus, the compounds of the present invention may be considered to be bactericidal and / or preferably bacteriostatic agents.

[0106] Such antibacterial properties may also enable the prevention of the onset of such infection or disease, the protection of cells in a host from (e.g., further) bacterial infection, the prevention or arrest of the spread of bacterial infection or disease (within a single host or from one host to a new host), or the prevention of bacterial reactivation after latency in the host.

[0107] According to a further aspect of the present invention there is provided a method for treating a bacterial infection which comprises administering to a patient in need of such treatment a compound of the present invention, or a salt thereof.

[0108] The compounds of the present invention may further have anti-cancer properties that may allow for the treatment of cancer itself, i.e., by interfering with cancer, as opposed to treating any symptoms of cancer, such as pain and / or inflammation. Such anti-cancer properties may also include the prevention of the onset of such diseases, for example, by treating inflammation and thereby preventing such onset.

[0109] According to another aspect of the present invention, there is provided a method for treating cancer, comprising administering to a patient in need of such treatment a compound of the present invention, or a salt thereof.

[0110] Specific cancers that may be mentioned include oral cavity cancer, nasopharyngeal cancer, middle ear cancer, conjunctival cancer, throat cancer, tracheal cancer, esophageal cancer, stomach cancer, intestinal cancer, cervical cancer, endometrial cancer, skin cancer caused by oral mucositis, rhinitis, otitis media, conjunctivitis, pharyngitis, laryngitis, tracheitis, esophagitis, gastritis, enterocolitis, cervicitis, endometritis, erythematous skin lesions, etc. A specific skin cancer that may be mentioned is basal cell carcinoma.

[0111] "Patient" includes reptilian, avian, and preferably mammalian (especially human) patients.

[0112] According to the present invention, the compounds of the present invention are administered in the form of a pharmaceutical preparation comprising the compound(s) in a pharmaceutically acceptable dosage form(s), preferably locally or systemically, for example orally, intravenously, or intra-arterially (including intravascular and other perivascular devices / dosage forms (e.g., stents)), intramuscularly, cutaneously, subcutaneously, transmucosally (e.g., sublingually or bucally), rectally, intravaginally, transdermally, intranasally, pulmonary (e.g., trachea or bronchus), preferably topically, or by any other parenteral route.

[0113] Administration by inhalation (eg, intranasal) is particularly useful when the condition being treated is rhinitis or inflammation resulting from a viral infection of the respiratory tract (eg, upper respiratory tract infections such as colds and influenza).

[0114] Pulmonary administration is particularly useful when the condition being treated is COPD or IPF. Local administration forms can be enhanced by, for example, using powder aerosols or by creating a spray containing the active ingredient by using an aqueous mist using suitable atomizing technology or device, such as a nebulizer.

[0115] Anorectal administration is particularly useful when the condition being treated is hemorrhoids or ulcerative colitis, using an appropriate delivery means such as an injectable foam solution or suppository.

[0116] Administration to the lower gastrointestinal tract can also be achieved by parenteral, particularly oral delivery, by using standard delayed-release or sustained-release coating technology known to those skilled in the art.Particularly, separate parts of the upper or lower intestine can be targeted.For example, colonic administration can also be achieved by colon-targeted drug delivery means that is first administered orally or parenterally.

[0117] Preferred modes of delivery of the compounds of the invention include local delivery to the site of inflammation (e.g., mucosa including oral and / or nasal mucosa, lung, anorectal and / or colon), or more preferably, skin), in a suitable (e.g., pharmaceutically and topically acceptable) vehicle, and / or a commercially available formulation, suitable for application to the skin and / or suitable mucosal surface, but may also include oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal, or pulmonary delivery.

[0118] The compounds of the present invention will generally be administered in the form of one or more, e.g., pharmaceutical formulations, mixed with (e.g., pharmaceutically acceptable) adjuvants, diluents, or carriers, which may be selected with due consideration of the intended route of administration (e.g., topical administration to relevant mucous membranes (including the lungs) or preferably to the skin) and standard pharmaceutical or other (e.g., cosmetic) practice. Such pharmaceutically acceptable carriers may be chemically inert to the active compounds and may be free of adverse side effects or toxicity under the conditions of use. Such pharmaceutically acceptable carriers may also provide immediate or modified release of the active ingredient.

[0119] Suitable pharmaceutical formulations are commercially available or can be found in the literature, for example, Remington The Science and Practice of Pharmacy, 22 nd edition, Pharmaceutical Press (2012), and Martindale-The Complete Drug Reference, 38 thEdition, Pharmaceutical Press (2014), as well as documents referenced therein, the relevant disclosures of all of which documents are incorporated herein by reference. Otherwise, the preparation of suitable formulations containing the compounds of the present invention can be achieved in ways not of the present invention by those skilled in the art using conventional techniques.

[0120] The compounds of the present invention may be in the form of an aqueous formulation such as an emulsion, suspension, and / or solution (e.g., an (optionally) buffered aqueous formulation (e.g., solution) such as a saline-containing formulation (e.g., solution), a phosphate-containing formulation (e.g., solution), an acetate-containing formulation (e.g., solution), or a borate-containing formulation (e.g., solution), or a lyophilized powder.

[0121] The active ingredient may additionally and / or alternatively be combined with suitable excipients to prepare: gel formulations (for which suitable gel matrix materials include cellulose derivatives, carbomers and alginates, tragacanth gum, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharides, sugars such as glucose, glycerin, propanediol, vinyl polymers, acrylic resins, polyvinyl alcohol, carboxyvinyl polymers, and, in particular, hyaluronic acid), Lotions (suitable matrix materials for which include cellulose derivatives, glycerin, non-cellulose polysaccharides, polyethylene glycols of different molecular weights, and propanediol), Pastes or ointments (in this regard, suitable paste matrix materials include glycerin, petrolatum, paraffin, polyethylene glycols of various molecular weights, etc.); Creams or foams (suitable excipients (e.g., foaming agents) in this regard include hydroxypropyl methylcellulose, gelatin, polyethylene glycol of various molecular weights, sodium dodecyl sulfate, sodium fatty alcohol polyoxyethylene ether sulfonate, corn gluten powder, and acrylamide); powder aerosols (suitable excipients for which include mannitol, glycine, dextrin, dextrose, sucrose, lactose, sorbitol, and polysorbates, e.g., dry powder inhalants); and / or Oral or inhaled liquids, e.g., water (aerosol) sprays (suitable excipients for these include viscosity regulators such as hyaluronic acid, sugars such as glucose and lactose, emulsifiers, buffers, alcohol, water, preservatives, sweeteners, flavors, etc.). Injection solutions or suspensions (which may be aqueous or other, for which suitable excipients include solvents and co-solvents, solubilizers, wetting agents, suspending agents, emulsifying agents, thickening agents, chelating agents, antioxidants, reducing agents, antimicrobial preservatives, buffers and / or pH adjusters, bulking agents, protectants, and tonicity modifiers).

[0122] Moisturizing agents such as glycerol, glycerin, polyethylene glycol, trehalose, glycerol, petrolatum, paraffin oil, silicone oil, hyaluronic acid and its salts (for example, sodium salt and potassium salt), octanoic acid / capric acid triglyceride, and / or antioxidants such as vitamins and glutathione, and / or pH adjusters such as acids, bases, and pH buffers may also be included in such formulations as needed.In addition, hexadecanol (cetyl alcohol), fatty acids (for example, stearic acid), sodium dodecyl sulfate (sodium lauryl sulfate), sorbitan esters (for example, sorbitan stearate, sorbitan oleate, etc.), monoacylglycerides (glyceryl monostearate, etc.), polyethoxylated alcohols, polyvinyl alcohols, polyol esters, polyoxyethylene alkyl ethers (for example, polyoxyethylene sorbitan monooleate), polyoxyethylene castor oil derivatives, ethoxylated fatty acid esters, Polyoxylglycerides, lauryldimethylamine oxide, bile salts (e.g., sodium deoxycholate, sodium cholate), phospholipids, N,N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide, poloxamer, lecithin, sterols (e.g., cholesterol), sugar esters, surfactants / emulsifiers such as polysorbates, preservatives such as phenoxyethanol and ethylhexylglycerin, and thickeners such as acryloyldimethyltaurate / VP copolymer may also be included. In particular, stearic acid, glyceryl monostearate, hexadecanol, sorbitan stearate, cetyl alcohol, octanoic / capric glycerides, etc. may be included, especially in cream formulations.

[0123] The compounds of the present invention and (e.g., pharmaceutical) formulations containing them (e.g., solutions, gels, creams, ointments, lotions, foams, pastes, and / or dry powders as described above) can be further combined with a suitable matrix material to prepare bandages or therapeutic patches for application to biological surfaces such as skin or mucosal surfaces. Thus, such formulations can be used to impregnate matrix materials such as gauze, nonwoven fabrics, or silk paper. Alternatively, the therapeutic patch can be, for example, a bandage, a face mask, an eye mask, a hand mask, a foot mask, etc.

[0124] Although petrolatum may be used for use in applying such dressings to wounds, we have also found that a PEG (e.g., PEG 400) based ointment can be combined with a matrix material to prepare a dressing without the need for petrolatum.

[0125] The compounds of the present invention can be administered for inhalation by suspension, dry powder, or solution. Suitable inhalation devices include pressurized metered dose inhalers (pMDIs), which can be manually or breath-activated and can be used with or without a standard spacer device; dry powder inhalers (DPIs), which can be single-dose, multi-dose, power-assisted; and soft mist inhalers (SMIs) or nebulizers, which deliver aerosolized medication in a fine mist at a slower rate than the spray delivered using, for example, a pMDI.

[0126] In pMDIs, the compounds of the invention may be administered as a pressurized suspension of micronized particles dispersed in a propellant (e.g., with an excipient such as HFA, mannitol, lactose, sorbitol, etc.) or as an ethanol solution to deliver one or more metered doses of about 20 to about 100 μL with each actuation. Actuation can be by hand (e.g., pushing) or by inhalation (breath actuation) and involves a spring-driven float-trigger system.

[0127] In DPIs, the compounds of the invention may be administered in the form of micronized drug particles (between about 1 and about 5 μm in size), either alone or mixed with larger particle-sized inert excipients (e.g., mannitol), in capsules that may be preloaded or manually filled into the device. Inhalation from a DPI may break down the drug particles and allow them to disperse in the respiratory tract.

[0128] In an SMI, the compound of the invention can be stored as a solution in a cartridge that is loaded into the device. A spring can release the dose into a micropump so that the dose is released when a button is pressed, releasing a jet of drug solution.

[0129] Various nebulizers can also be used to administer the compounds of the present invention in the form of a fine mist of aerosolized solution, including exhalation-enhanced jet nebulizers (with the aid of a compressor, airflow moves through a jet, aerosolizing the medicinal solution), exhalation-activated jet nebulizers (with the aid of a compressor, airflow moves through a tube, aerosolizing the medicinal solution after the patient inhales), ultrasonic nebulizers (piezoelectric crystals vibrate and heat to cause aerosolization, resulting in nebulization), and vibrating mesh nebulizers (piezoelectric crystals vibrate mesh plates to cause aerosolization, generating very fine droplets without significantly changing the temperature of the solution during nebulization).

[0130] According to a further aspect of the present invention there is provided a process for the preparation of a pharmaceutical composition / formulation as defined herein, which process comprises bringing into association a compound of the invention as defined above with one or more pharmaceutically acceptable excipients as defined above.

[0131] The compounds of the present invention may also be combined in treatment with one or more growth factors selected from platelet-type growth factors (including platelet-derived growth factor, PDGF), osteosarcoma-derived growth factor (ODGF), epidermal growth factor (EGF), transforming growth factors (TGFα and TGFβ), fibroblast growth factors (αFGF, βFGF), insulin-like growth factors (IGF-I, IGF-II), nerve growth factor (NGF), interleukin-type growth factors (IL-1, IL-1, IL-3), erythropoietin (EPO), and colony-stimulating factors (CSF).

[0132] According to a further aspect of the present invention, there is provided a (e.g., pharmaceutical) composition comprising a compound of the present invention and one or more pharmaceutically acceptable excipients, such as an adjuvant, diluent, or carrier. Preferred formulations are suitable for topical application, for example, to mucosa (including the oral and / or nasal mucosa, lung, anorectal and / or colon), or more preferably, to the skin, and therefore include a topically acceptable adjuvant, diluent, or carrier.

[0133] Thus, there is further provided a pharmaceutical composition comprising a compound of the invention that is suitable, adapted, and / or packaged and presented for topical administration (e.g., to the oral and / or nasal mucosa, lungs, anorectal region and / or colon, or preferably to the skin), and the use of such a formulation in the treatment of disorders, including inflammation, inflammatory disorders, and / or conditions (e.g., as a symptom) characterized by inflammation, by direct topical administration of the formulation (e.g., to the mucosa, including the oral and / or nasal mucosa, lungs, anorectal region and / or colon, or preferably to the skin).

[0134] For the avoidance of doubt, with regard to this aspect of the present invention, topical formulations comprising the compounds of the present invention can be used in the treatment of any and all inflammatory disorders(ies) and / or in the treatment of any and all conditions characterized by inflammation as mentioned, defined, or described herein above, in any and all conditions described herein, including the treatment of inflammation.Similarly, the topical formulations comprising the compounds of the present invention that may be mentioned include any and all of those mentioned, defined, or described above.Any and all of the relevant disclosures herein are incorporated herein by reference in connection with this aspect of the present invention.

[0135] Topical formulations (e.g., liquid or (e.g., aqueous) solution-based) containing the compounds of the present invention may be particularly useful in wound healing and may alleviate pain (including soreness) and pruritus / itch, particularly associated with the wound itself and the wound healing process. Such topical formulations containing the compounds of the present invention may be particularly useful in preventing and / or suppressing exudation of fluid from the wound after a burn or wound, especially during the acute inflammatory phase, e.g., the first 48 hours. This prevents the risk of infection and other physiological reactions. Such topical formulations containing the compounds of the present invention may also be particularly useful in preventing and / or suppressing scarring and melanin pigmentation (see above), whether or not associated with a wound.

[0136] The administration of the active ingredient can be continuous or intermittent. The mode of administration can also be determined by the timing and frequency of administration, and in the case of therapeutic treatment of inflammation, by the severity of the condition.

[0137] Depending on the disorder and patient being treated, as well as the route of administration, the compounds of the present invention may be administered to a patient in need of treatment in different therapeutically effective doses.

[0138] Similarly, the amount of active ingredient in the formulation will depend on the severity of the condition being treated and the patient, but may be determined by one skilled in the art.

[0139] In any event, a practitioner or other skilled artisan can routinely determine the actual dosage that will be most suitable for an individual patient, depending on the severity of the condition and the route of administration. The dosages mentioned herein are exemplary of the average case, and there can, of course, be individual instances in which higher or lower dosage ranges are merited, and these are within the scope of this invention.

[0140] The dose can be administered one to four (eg, three) times daily.

[0141] Suitable concentrations of the compounds of the invention in the aqueous solution product can be from about 0.01 (eg, about 0.1) to about 15.0 mg / mL, in all cases calculated as the free (non-salt) peptide.

[0142] A suitable topical dose of the compounds of the invention is about 5 μg / cm, in all cases calculated as the free (non-salt) compound. 2 Approximately 1 to 10 μg / cm of the treatment area 2 ) treatment area, about 0.1 (e.g., about 0.5) to about 20 μg / cm 2 Approximately 0.05 to approximately 50 μg / cm of treatment area 2 within the therapeutic range of

[0143] Suitable doses of the compounds of the present invention for nasal administration (e.g., by inhalation) range from about 0.01 μg to about 2000 mg, for example, from about 0.1 μg to about 500 mg, or from 1 μg to about 100 mg. Specific doses for nasal administration that may be mentioned include doses of about 10 μg to about 1 mg, particularly about 0.1 mg (i.e., about 100 μg). Nasal administration of about 0.1 mg of the compounds of the present invention per day has been found to be particularly effective in treating conditions associated with inflammation of the nasal cavity and mucous membranes, such as rhinitis (e.g., allergic rhinitis).

[0144] Suitable doses of the compounds of the invention for pulmonary administration (e.g., by inhalation) range from about 0.01 μg to about 2000 mg, for example, from about 0.1 μg to about 500 mg, or from 1 μg to about 100 mg. Particular doses for pulmonary administration that may be mentioned include doses from about 10 μg to about 10 mg, in particular doses from about 0.6 mg (i.e., 60 μg) to 6 mg (e.g., for use in the treatment of COPD or idiopathic pulmonary fibrosis).

[0145] The pH value of a formulation containing the compound of the present invention is preferably within the range of about 1.0 to about 9.0 (for example, about 3.0 to about 8.0).

[0146] In any case, the dose administered to a mammal, particularly a human, in the context of the present invention must be sufficient to bring about a therapeutic response in the mammal over a reasonable time frame (as described above).Those skilled in the art will recognize that the selection of the exact dose and composition and the most appropriate delivery regimen will also be influenced by, among other things, the pharmacological properties of the formulation, the nature and severity of the condition being treated, the physical and mental state of the recipient, and the age, condition, weight, sex and response of the patient being treated, the stage / severity of the disease, and genetic differences between patients.

[0147] In the uses and methods described herein, the compounds of the present invention may be combined with one or more active ingredients (other anti-inflammatory agents) useful in the treatment of inflammation and / or inflammatory disorders. Accordingly, such patients may also (and / or already) be undergoing a therapy based on the administration of one or more of such other active ingredients, by which is meant receiving prescribed doses of one or more of the active ingredients mentioned herein before, in addition to, and / or after treatment with the compounds of the present invention.

[0148] The anti-inflammatory agent that can be used in combination with the compound of the present invention in the treatment of inflammation includes the therapeutic agent that is useful for treating inflammation and / or the disease characterized by inflammation as one of its symptoms.Depending on the pathological condition to be treated, such anti-inflammatory agent can also include NSAIDs, leukotriene receptor antagonists (for example, montelukast itself), corticosteroids, analgesics, and certain enzymes such as trypsin, as described below.The compound of the present invention can also be combined with leukotriene B4 (LTB4).

[0149] In this context, the compounds of the invention may also be combined with one or more mussel adhesive proteins (MAPs) for use in treating inflammation, including any adhesive protein that may be derived from a mussel species such as Mytilus edulis (blue mussel), including full-length proteins including all subtypes that are or may be derived from mussels, such as collagens pre-COL-P, pre-COL-D, and pre-COL-NG, mussel foot matrix proteins PTMP and DTMP, and more preferably mfps or mefps (such as mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, and especially mefp-1), including mixtures or combinations of any of these proteins, such as mefps. Naturally occurring MAP can be prepared, for example, by mixed adsorption chromatography (see Chinese Patent No. ZL200710179491.0), by carboxymethyl ion exchange chromatography (see Chinese Patent No. ZL200710179492.5), and / or by salting out and dialysis (Chinese Patent No. ZL200910087567.6). Commercial sources of MAP include USUN Bio Co. (China, sold as MAP Medical Device®), BD Biosciences (USA), Kollodis (Korea), and Biopolymer (Sweden). Alternatively, MAP may be produced using known recombinant DNA methods.

[0150] Derivatives (e.g., pharmaceutically acceptable derivatives) of MAPs may be combined with the compounds of the invention, including, for example, compounds having molecular weights ranging from about 500 Da to about 2,000 Da (e.g., about 1,500, such as about 1,200, including about 800 Da). Such derivatives may also include other compounds containing amino acid sequences that are the same as or are (e.g., minor) variants (as defined above) of sequences identified in naturally occurring MAPs, which can be synthesized by chemical and / or biological processes (e.g., chemical modification of naturally occurring MAPs or direct synthesis).

[0151] For example, as discussed herein above, the sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (mefp-1 decapeptide, SEQ ID NO: 4), and Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 12) The isolated decapeptide compounds of are pharmaceutically acceptable low molecular weight derivatives of MAP that can be combined with the compounds of the present invention.

[0152] Other preferred agents that can be combined with the compounds of the invention include LTB4 (for treating wounds and burns), montelukast (for treating inflammation generally), and trypsin (for treating inflammation of the mucous membranes associated with, for example, viral infections).

[0153] The compounds of the present invention may also be combined with other therapeutic agents that are known to cause inflammation as a side effect when administered.

[0154] When the compounds of the invention are "combined" with other therapeutic agents in this manner, the active ingredients may be administered together in the same formulation or separately (simultaneously or sequentially) in different formulations.

[0155] Such combination products provide for the administration of a compound of the invention in combination with another therapeutic agent, and therefore may be presented as separate formulations (at least one of which contains a compound of the invention and at least one of which contains the other therapeutic agent) or may be presented (i.e., formulated) as a combined preparation (i.e., presented as a single formulation containing a compound of the invention and the other therapeutic agent).

[0156] therefore, (1) A pharmaceutical formulation comprising a compound of the present invention, another anti-inflammatory agent or a drug known to cause inflammation as a side effect, and a pharmaceutically acceptable excipient (e.g., an adjuvant, diluent, or carrier) (this formulation is hereinafter referred to as a "combination preparation"); and (2) A parts kit comprising: (A) a pharmaceutical formulation comprising a compound of the present invention mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier; (B) a pharmaceutical formulation containing another anti-inflammatory agent, or an agent known to cause inflammation as a side effect, mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier; There is further provided a kit of parts, wherein components (A) and (B) are each provided in a form suitable for administration in combination with the other.

[0157] In a further aspect of the present invention, there is provided a process for preparing a combination preparation as defined above, which comprises associating a compound of the present invention, another anti-inflammatory agent, or an agent known to cause inflammation as a side effect, with at least one (e.g., pharmaceutically acceptable) excipient.

[0158] In a further aspect of the present invention, there is provided a process for preparing a kit-of-parts as defined herein above, comprising associating components (A) and (B). As used herein, reference to associating may mean that the two components are suitable for administration in combination with each other.

[0159] Thus, by "associating" two components with each other, with respect to the process for the preparation of a kit-of-parts as defined herein above, we mean that the two components of the kit-of-parts: (i) may be provided as separate formulations (i.e., independently of each other) and then combined for use in conjunction with each other in combination therapy; or (ii) They may be packaged and presented together as separate components of a "combination pack" for use in conjunction with each other in combination therapy.

[0160] therefore, (I) one of components (A) and (B) as defined herein, (II) A kit of parts is further provided that includes the component together with instructions for use in combination with the other of the two components.

[0161] The kits of parts described herein may contain two or more formulations containing suitable amounts / doses of a compound of the invention and / or two or more formulations containing suitable amounts / doses of another anti-inflammatory agent to provide for repeated administration. When two or more formulations (containing any of the active compounds) are present, such formulations may be the same or different with respect to the dose, chemical composition(s), and / or physical form(s) of any of the compounds.

[0162] With respect to the kits of parts described herein, by "administration in combination with," we include sequential, separate, and / or simultaneous administration of respective formulations comprising a compound of the invention and another anti-inflammatory agent over the course of treatment of the relevant condition.

[0163] Thus, with respect to a combination product according to the invention, the term "administration in combination with" includes administration of the two components of the combination product (a compound of the invention and another anti-inflammatory agent) either together or sufficiently closely in time (optionally repeatedly) so as to permit a greater beneficial effect to the patient over the course of treatment of the relevant conditions than would occur if, over the same course of treatment, either a formulation comprising the compound of the invention or a formulation comprising the other agent were administered alone (optionally repeatedly) without the other component. Determining whether a combination provides a greater beneficial effect with respect to the treatment of a particular condition, and over the course of treatment, will depend on the condition being treated or prevented, but can be routinely accomplished by one of ordinary skill in the art.

[0164] Furthermore, in the context of the kit-of-parts according to the invention, the term "in combination with" includes that one or the other of the two formulations may be administered before, after, and / or simultaneously (optionally repeatedly) with the administration of the other component. When used in this context, the terms "co-administered" and "administered simultaneously with" include administration of individual doses of the relevant compound of the invention and the other anti-inflammatory agent within 48 hours (e.g., 24 hours) of each other.

[0165] Whenever the term "about" is used herein in the context of quantities such as concentration and / or dosage of an active ingredient, molecular weight, or pH, it will be understood that such variables are approximate and thus may vary from the numerical values ​​specified herein by ±10%, e.g., ±5%, and preferably ±2% (e.g., ±1%). In this regard, the term "about 10%" means, for example, ±10% of the numerical value 10, i.e., 9% to 11%.

[0166] The compounds of the present invention have the advantage that they can be used in a variety of conditions characterized by inflammation, whether the condition itself is an organic inflammatory disease or is associated with or characterized by inflammation (e.g., wounds, burns, or viral infections).

[0167] The uses and methods described herein may also have advantages over similar compounds or methods (treatments) known in the prior art in treating the above-mentioned conditions, whether for use in treating inflammation, inflammatory disorders, or disorders characterized by inflammation as a symptom (including wounds), or other methods, in that they may be more convenient for the physician and / or patient, more effective, less toxic, have a broad spectrum of activity, be more potent, cause fewer side effects, or have other useful pharmacological properties over similar methods (treatments).

[0168] The present invention is illustrated by the following examples, in which for various compounds, including the compounds of the present invention, Figure 1 shows the swelling rate in a mouse ear swelling model, Figure 2 shows the Hyp content (and therefore the recovery level), Figure 3 shows the vascular endothelial growth factor and transforming growth factor-beta 1 levels of wound tissue in an acute wound mouse model, Figure 4 shows the rate of unhealed wounds in a diabetic wound mouse model, Figure 5 shows skin regeneration, Figure 6 shows the fibroblast proliferation score, Figure 7 shows the ratio of remaining wound area compared to the initial wound, and Figures 8-11 show various markers of wound healing (skin regeneration, fibrogenic proliferation, inflammation, and inflammatory processes, respectively). FIG. 12 shows the results of histopathological analysis for edema levels in different groups in a further diabetic wound model; FIGS. 13 and 17 both show the effect of compounds of the invention on edema caused by acute inflammation in a further mouse ear swelling model; FIG. 14 shows the rate of unhealed wounds in a further acute wound mouse model; FIG. 15 shows a comparison between compounds of the invention and known anti-inflammatory steroids in a mouse ear swelling model; and FIG. 16 shows the IL-1β content of FIG. 1 in lung tissue for different compounds of the invention in a mouse lung injury model. [Brief explanation of the drawings]

[0169] [Figure 1] 1 shows the swelling rate in a mouse ear swelling model. [Figure 2]Hyp content (and therefore recovery level) is indicated. [Figure 3] FIG. 1 shows vascular endothelial growth factor and transforming growth factor-beta 1 levels in wound tissue in an acute wound mouse model. [Figure 4] 1 shows the rate of non-healing wounds in a diabetic wound mouse model. [Figure 5] Shows skin regeneration. [Figure 6] Fibroblast proliferation scores are shown. [Figure 7] The ratio of the remaining wound area compared to the initial wound is shown. [Figure 8] 1 shows the results of histopathological analysis for various markers of wound healing. [Figure 9] 1 shows the results of histopathological analysis for various markers of wound healing. [Figure 10] 1 shows the results of histopathological analysis for various markers of wound healing. [Figure 11] 1 shows the results of histopathological analysis for various markers of wound healing. [Figure 12] 10 shows the level of edema in different groups in a further diabetic wound model. [Figure 13] Figure 1 shows the effect of compounds of the invention on edema caused by acute inflammation in a further mouse ear swelling model. [Figure 14] 1 shows the rate of non-healing wounds in a further acute wound mouse model. [Figure 15] 1 shows a comparison between compounds of the present invention and known anti-inflammatory steroids in a mouse ear swelling model. [Figure 16] FIG. 1 shows the IL-1β content in lung tissue of different compounds of the present invention in a mouse lung injury model. [Figure 17] Figure 1 shows the effect of compounds of the invention on edema caused by acute inflammation in a further mouse ear swelling model. [Example]

[0170] Example 1 Synthesis of montelukast styrene-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (i.e., montelukast styrene covalently bound at the N-terminus to amino acid SEQ ID NO: 12) To synthesize 3 mmol of peptide SEQ ID NO: 12, the following procedure was followed.

[0171] Fmoc-Lys-Boc-Wang resin (9.15 g, GLS180322-41301, GL Biochem, Shanghai, China) was packed into a glass reaction column.

[0172] Methylene chloride (DCM, 200 mL, Shandong Jinling Chemical Industry Co Ltd., Shandong, China) was added to the column and the resin was allowed to soak for approximately 30 minutes. Then, the DCM was removed by vacuum filtration.

[0173] The resin was washed three times with N,N-dimethylformamide (DMF, 200 mL, Shandong Shitaifeng Fertilizer Industry Co Ltd, Shandong, China).

[0174] A 20% piperidine solution in DMF (200 mL, Shandong Shitaifeng Fertilizer Industry Co Ltd, Shandong, China) was added as a deprotection solution and reacted for 20 min. Then, the solution was removed by vacuum filtration, and the column was washed with DMF six times.

[0175] Fmoc-Tyr(tBu)-OH (4.14 g, GLS170916-36901, GL Biochem, Shanghai, China) and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylaminium tetrafluoroborate (TBTU, 2.89 g, GLS170805-00705, GL Biochem, Shanghai, China) were added to the resin. DMF (150 mL) was added to the reaction column, followed by N,N-diisopropylethylamine (DIPEA, 2.33 g, Suzhou Highfine Biotech Co. Ltd., Jiangsu, China). A color reaction was detected on the resin after 30 min, indicating the reaction was complete. The solvent was removed by vacuum filtration.

[0176] The above coupling steps were repeated to couple the remaining amino acids in the same amounts (in moles): Fmoc-Thr(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH, and Fmoc-Ala-OH.

[0177] Finally, montelukast (5.47 g, MedChemExpress, MCE China, Shanghai, China) was added to the resin. Then, the liquid was drained after 15 min, and the column was washed with DMF, DCM, and methanol three times each.

[0178] A 91.5 mL (i.e., 10 mL per gram of resin) lysate consisting of 95% trifluoroacetic acid (TFA), 2.5% water, and 2.5% triisopropylsilane (Tis) was added to soak the resin-bound peptide-containing compound. The side chains were also deprotected during cleavage. After cleavage, the solid support was removed by filtration, and the filtrate was concentrated under reduced pressure. The cleaved peptide was precipitated with diethyl ether and lyophilized to yield 600 mg of crude title compound.

[0179] 1 mg of the crude product was dissolved in 1 mL of a 1:3 mixture of acetonitrile and water and detected using a P3000A HPLC pump and an LC3000 semi-preparative instrument (preparative column model: GS-120-10-C18-AP 30 mm, Beijing Chuangxin Tonggheng Science & Technology Co., Ltd., Beijing, China). A suitable gradient was calculated for elution, and the target peak was detected by LCMS at 11:035 min (analytical column model: GS-120-5-C18-BIO, 4.6 × 250 mm, detection: UV at 220 nm, solvent A: 0.1% TFA in MeCN, solvent B: 0.1% TFA in water, flow rate: 1.0 mL / min, volume: 10 μL).

[0180] The crude compound was desalted using an anion exchange resin, analyzed, and lyophilized. After purification, approximately 50 mg of purified peptide was obtained, which was retested for confirmation.

[0181] MS: m / z 866.90[M+2H] 2+ .

[0182] Based on the characterization data available and presented herein, it is understood that the compound prepared in this example is the one identified above as the title compound. Otherwise, the compound prepared in Example 1 is a compound of the invention in which n is 0 in a compound of Formula I and the compound of Formula I is covalently attached at the N-terminus of amino acid SEQ ID NO: 12. In any event, the compound of Example 1 is hereinafter referred to as "Compound A."

[0183] Example 2 Synthesis of Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (montelukast styrene) (i.e., montelukast styrene covalently linked to amino acid SEQ ID NO: 4 at the C-terminal Lys) A similar procedure to that described in Example 1 above was used starting with Fmoc-Lys(Dde)-OH (CAS number: 150629-67-7) on Wang resin.

[0184] A 25% solution of piperidine in DMF (200 mL, Shandong Shitaifeng Fertilizer Industry Co Ltd, Shandong, China) was added to remove the protecting Fmoc group.

[0185] The second protected amino acid Fmoc-DOPA(acetonide)-OH was added along with TBTU and DIPEA until the reaction was complete.

[0186] The above coupling steps were repeated to couple the remaining amino acids in the same amounts (in moles): Fmoc-Thr(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH, and Fmoc-Ala-OH.

[0187] The peptidyl resin was placed in a flask and treated with 2% hydrazine monohydrate (25 mL / g) in DMF. The flask was stoppered, and the mixture was allowed to stand at room temperature for 3 minutes. The resin was then washed with DMF. Montelukast (5.47 g, MedChemExpress, MCE China, Shanghai, China) was added to the resin along with TBTU and DIPEA, and the mixture was allowed to react for 1 hour.

[0188] The protected peptidyl resin was treated with 91.5 mL of lysate (a mixture of 95% TFA, 2.5% water, and 2.5% Tis) for 1 hour. After cleavage, the solid support was removed by filtration, and the filtrate was concentrated under reduced pressure. The cleaved peptide was precipitated with diethyl ether and lyophilized to give approximately 600 mg of the crude title compound.

[0189] After purification, 50 mg of pure product was obtained.

[0190] MS: m / z 875.75[M+2H] 2+ .

[0191] Based on the characterization data available and presented herein, it is understood that the compound prepared according to this example is the one identified above as the title compound. Otherwise, the compound prepared in Example 2 is a compound of the invention in which n is 0 in the compound of Formula I and the compound of Formula I is covalently attached to amino acid SEQ ID NO: 4 on the C-terminal Lys. In any event, the compound of Example 2 is hereinafter referred to as "Compound B."

[0192] Example 3 Synthesis of montelukast styrene-Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr-Lys (i.e., montelukast styrene covalently bound at the N-terminus to amino acid SEQ ID NO: 13) A procedure similar to that described in Example 4 below was used starting with Fmoc-Lys(Dde)-OH (CAS number: 150629-67-7) on Wang resin.

[0193] A 25% solution of piperidine in DMF (200 mL, Shandong Shitaifeng Fertilizer Industry Co Ltd, Shandong, China) was added to remove the protecting Fmoc group.

[0194] The second protected amino acid Fmoc-Tyr(tBu)-OH (GLS170916-36901, GL Biochem, Shanghai, China) was added along with ByBOP and DIPEA until the reaction was complete.

[0195] The above coupling steps were repeated to couple the remaining amino acids in the same amounts (in moles): Fmoc-Thr(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-pSer(tBu)-OH, Fmoc-Pro-OH, Fmoc-Lys(Boc)-OH, and Fmoc-Ala-OH.

[0196] The peptidyl resin was placed in a flask and treated with 2% hydrazine monohydrate (25 mL / g) in DMF. The flask was stoppered, and the mixture was allowed to stand at room temperature for 3 minutes. The resin was then washed with DMF. Montelukast (1.8 g, MedChemExpress, MCE China, Shanghai, China) was added to the resin along with TBTU and DIPEA, and the mixture was allowed to react for 1 hour.

[0197] A lysate (10 mL per gram of resin) consisting of 95% trifluoroacetic acid (TFA), 2.5% water, and 2.5% triisopropylsilane (Tis) was added to soak the resin-bound peptide-containing compound. The side chains were also deprotected during cleavage. After cleavage, the solid support was removed by filtration, and the filtrate was concentrated under reduced pressure. The cleaved peptide was precipitated with diethyl ether and lyophilized to yield 1.4 g of crude peptide.

[0198] 1 mg of crude product was dissolved in 1 mL of a mixture of acetonitrile and water (1:3) and detected using a P3000A HPLC pump and an LC3000 semi-preparative instrument (preparative column model: GS-120-10-C18-AP 30 mm, Beijing Chuangxintongheng Science & Technology Co., Ltd., Beijing, China). A suitable gradient was calculated for elution, and the target peak was detected by LCMS (analytical column model: GS-120-5-C18-BIO, 4.6*250 mm).

[0199] The crude compound was desalted using an anion exchange resin, analyzed, lyophilized, and retested for confirmation.

[0200] After purification, 98 mg of pure product was obtained (approximately 7% yield from crude product).

[0201] MS: m / z 907.3 [M+2H] 2+ .

[0202] Based on the characterization data available and presented herein, it is understood that the compound prepared in this example is the one identified above as the title compound. Otherwise, the compound prepared in Example 3 is a compound of the invention in which n is 0 in a compound of Formula I and the compound of Formula I is covalently attached at the N-terminus of amino acid SEQ ID NO: 13. In any event, the compound of Example 3 is hereinafter referred to as "Compound C."

[0203] Example 4 Synthesis of Ala-Lys(montelukast styrene)-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys(montelukast styrene) (i.e., two molecules of montelukast styrene covalently linked to amino acid SEQ ID NO: 12 via Lys residues). To synthesize 1 mmol of peptide SEQ ID NO: 12, the following procedure was followed.

[0204] Fmoc-Lys(Dde)-OH (CAS number: 150629-67-7) on Wang resin (3 g, GL Biochem, Shanghai, China) was packed into a glass reaction column.

[0205] Methylene chloride (DCM, 60 mL, Shandong Jinling Chemical Industry Co Ltd., Shandong, China) was added to the column and the resin was allowed to soak for approximately 30 minutes. Then, the DCM was removed by vacuum filtration.

[0206] The resin was washed three times with N,N-dimethylformamide (DMF, 60 mL, Shandong Shitaifeng Fertilizer Industry Co Ltd, Shandong, China).

[0207] A 20% piperidine solution in DMF (30 mL, Shandong Shitaifeng Fertilizer Industry Co Ltd, Shandong, China) was added as a deprotection solution and reacted for 20 min. Then, the solution was removed by vacuum filtration, and the column was washed with DMF six times.

[0208] Fmoc-Tyr(tBu)-OH (1.4 g, GLS170916-36901, GL Biochem, Shanghai, China) and benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (ByBOP, 1.56 g, Suzhou Highfine Biotech Co. Ltd, Jiangsu, China) were added to the resin, followed by DIPEA (1 mL, Suzhou Highfine Biotech Co. Ltd, Jiangsu, China). A color reaction was detected on the resin after 30 min, indicating the reaction was complete. The solvent was removed by evaporation.

[0209] The above coupling steps were repeated to couple the remaining amino acids in the same amounts (in moles): Fmoc-Thr(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-4-Hyp(tBu)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Lys(Dde)-OH, and Boc-Ala-OH.

[0210] The peptidyl resin was placed in a flask and treated with 2% hydrazine monohydrate (25 mL / g) in DMF. The flask was stoppered, and the mixture was allowed to stand at room temperature for 3 minutes. The resin was then washed with DMF. Montelukast (3.6 g, MedChemExpress, MCE China, Shanghai, China) was added to the resin along with TBTU and DIPEA, and the mixture was allowed to react for 1 hour.

[0211] A lysate (30 mL, 10 mL per gram of resin) consisting of 95% trifluoroacetic acid (TFA), 2.5% water, and 2.5% triisopropylsilane (Tis) was added to soak the resin-bound peptide-containing compound. The side chains were also deprotected during cleavage. After cleavage, the solid support was removed by filtration, and the filtrate was concentrated under reduced pressure. The cleaved peptide was precipitated with diethyl ether and lyophilized to yield 1.6 g of crude peptide.

[0212] 1 mg of crude product was dissolved in 1 mL of a mixture of acetonitrile and water (1:3) and detected using a P3000A HPLC pump and an LC3000 semi-preparative instrument (preparative column model: GS-120-10-C18-AP 30 mm, Beijing Chuangxintongheng Science & Technology Co., Ltd., Beijing, China). A suitable gradient was calculated for elution, and the target peak was detected by LCMS (analytical column model: GS-120-5-C18-BIO, 4.6*250 mm).

[0213] The crude compound was desalted using an anion exchange resin, analyzed, lyophilized, and retested for confirmation. 1.6 g of crude peptide yielded 20 mg of purified peptide (90%-95% purity).

[0214] MS: m / z 762.2 [M+3H] 3+ .

[0215] Based on the characterization data available and presented herein, it is understood that the compound prepared in this example is the one identified above as the title compound. Otherwise, the compound prepared in Example 4 is a compound of the invention in which n is 0 in a compound of Formula I and the compound of Formula I is covalently attached at the Lys terminus of amino acid SEQ ID NO: 12. In any event, the compound of Example 4 is hereinafter referred to as "Compound D."

[0216] Example 5 Mouse ear swelling model I Thirty-five healthy male BALB / c mice, 6-8 weeks old and weighing an average of 18-25 g, were supplied by Changzhou Cvens Experimental Animal Co., Ltd. and housed and cared for for approximately one week prior to the experiment. The temperature of the enclosure was 25-27°C, humidity was 74%, and the mice were kept on a 12-hour alternating light-dark cycle with free access to food and water. The mice were randomly divided into seven groups, with five mice in each group, as described in Table 1 below.

[0217] The left ear of each mouse served as a control. The right ear of each mouse was treated with various different treatments, as summarized in Table 1 below. 20 μL of xylene (Shanghai Aladdin Bio-Chem Technology Co., Ltd., Shanghai, China) was applied to both the inside and outside of the right ear of each mouse. After approximately 4 minutes, the ear began to swell. Next, 0.08 g of each study treatment or vehicle was applied to the right ear of each group. The mice were then returned to their cages.

[0218] A montelukast sodium-based cream was made consisting of the following ingredients: montelukast sodium (200 mg, Arromax Pharmatech Co., Ltd., Suzhou, China), stearic acid (2 g), glyceryl monostearate (2 g), hexadecanol (2 g), glycerin (5 g), and sodium hydroxide (0.25 g) (all from Sinopharm Chemical Reagent Co., Ltd., Shanghai, China), ammonium acryloyldimethyltaurate / VP copolymer (0.13 g, Clariant Chemical (Guangzhou) Co., Ltd., Guangzhou, China), phenoxyethanol (0.3 g) and ethylhexylglycerin (0.1 g) (both from Shanghai Rayson Chemicals Co., Ltd., Shanghai, China), and purified water (88.42 g).

[0219] Stearic acid, glycerin monostearate, and hexadecanol were mixed and heated to 85°C with stirring until the mixture was completely dissolved. Ammonium acryloyldimethyltaurate / VP copolymer, purified water, and sodium hydroxide were mixed with stirring at 85°C to form a homogeneous colloidal suspension. Montelukast sodium, glycerin, phenoxyethanol, and ethylhexylglycerin were then mixed with stirring until the montelukast was completely dissolved.

[0220] The copolymer / water mixture was added to the stearic acid-containing mixture and emulsified by rapid stirring for 5 minutes using an emulsifier. The resulting emulsion was cooled to 55°C, and the montelukast-containing mixture was added with mixing. The resulting mixture was allowed to cool to room temperature to obtain the final product.

[0221] Dexamethasone cream (DEX) was made using the same procedure, except that montelukast was replaced with 0.4 mg of dexamethasone (Shanghai Aladdin Bio-Chem Technology Co., Ltd, Shanghai, China).

[0222] A gel containing Compound A ("A gel") was made consisting of the following components: 0.5 g of Compound A powder (obtained from GL Biochem, Shanghai, China, prepared as described in Example 1 above), methylcellulose (2.2 g, Shandong Guangda Technology Development Co., Ltd., Shandong Province, China), glycerin (11 g) and 11 g of propanediol (both from Sinopharm Chemical Reagent Co. Ltd.), and purified water (75.3 g).

[0223] Methylcellulose and water were mixed together and stirred until a uniform colloidal suspension was formed. Compound A powder, glycerin, and propanediol were then added to the methylcellulose / water mixture, and the resulting mixture was rapidly stirred for 5 minutes to obtain the final product.

[0224] A gel based on Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (Compound 1 (SEQ ID NO: 12), 1.5 g, obtained as a powder from GL Biochem, Shanghai, China, prepared by essentially the same process as described in Example 1 above, without the final coupling of montelukast) was prepared by the same process described above for Compound A ("1 gel").

[0225] Vehicle-1 in Table 1 below is a cream base that does not contain any active ingredients. Vehicle-2 in Table 1 below is a gel base that does not contain any active ingredients. Both were made using the same procedure as above, without the addition of any active ingredients. [Table 1]

[0226] Mice were sacrificed by cervical dislocation 40 minutes later. Both ears were amputated. Ear sections were collected from the same area on both ears using an 8 mm diameter skin pouch (Electron Microscopy Sciences, Hatfield, PA, USA). Weights were recorded, and the swelling rate was calculated as follows:

[0227] Swelling rate = (right ear weight - left ear weight) / left ear weight x 100% The results are shown in Table 2 below and in FIG. [Table 2]

[0228] The above results show that both test compounds reduce xylene-induced swelling.

[0229] Example 6 Acute Wound Model I Male C57BL / 6 mice, 6–8 weeks old, were supplied by Changzhou Cvens Experimental Animal Co. Ltd. Before the experiments were performed, the mice were housed under standardized conditions (constant temperature or 22 ± 2°C, alternating 12-hour light / dark cycles) and fed standard mouse chow containing water for approximately 1 week.

[0230] General anesthesia was induced using intraperitoneal 3% chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China, 1 mL / 10 g body weight). The hair on the back was shaved with a baby shaver and depilated with cream. The skin area was wiped and sterilized twice with 75% alcohol.

[0231] A single round wound was created in the midline of the back using an 18 mm diameter EMS skin biopsy punch (Electron Microscopy Sciences, PO Box 550, 1560 Industry Road, Hatfield, PA 19440). Full-thickness skin was removed, reaching the fascia. The wound was left open without sutures.

[0232] Different drugs were administered topically at 50 μL per wound once daily from day 0 to day 7. The model groups received the same amount of saline. As shown in Table 3 below, there were seven groups containing 56 mice in this experiment.

[0233] Recombinant human epidermal growth factor (rhEGF, Shanghai Haohai Biological Technology Co., Ltd., Shanghai, China) was purchased and prepared according to the manufacturer's instructions. Lyophilized rhEGF powder (100,000 IU / vial) was dissolved in 20 mL of saline to produce a solution with a concentration of 5,000 IU / mL. The working dose of rhEGF in this experiment was 1,285 IU / wound.

[0234] Compound B was obtained as a powder from GL Biochem and prepared as described in Example 2 above. Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (Compound 2, SEQ ID NO: 4) was obtained as a powder from GL Biochem and prepared essentially as described in Example 2 above, but without final coupling of montelukast. The powder was stored at -20°C and dissolved in saline at the concentrations shown in Table 3 below (L and H indicate low and high doses, respectively). [Table 3]

[0235] Hydroxyproline (Hyp) is a non-proteinogenic amino acid found in collagen, which contains approximately 12-14% Hyp by mass. Therefore, the Hyp content in tissue hydrolysates is a direct measure of the amount of collagen present, and the Hyp content presented in wound tissue directly indicates the level of recovery. The Hyp content of each group, as well as on days 4 and 7 after the start of treatment, are shown in Figure 2.

[0236] The results showed that the test compounds improved Hyp content in all treatment groups. Low doses of both Compound 2 and Compound B showed an early effect (day 4 (D4)). High doses of both test compounds also showed an accelerated effect on Hyp production on day 7 (D7).

[0237] Vascular endothelial growth factor (VEGF) and transforming growth factor-beta 1 (TGF-β1) play important roles in the wound healing process. VEGF and TGF-β1 are often co-expressed in tissues where angiogenesis occurs. The contents of these two factors in wound tissues have also been detected and are shown in Figure 3.

[0238] The results showed that the two peptides could stimulate the production of VEGF and TGF-β1.

[0239] Example 7 Diabetic Wound Model I Similar experiments using essentially the same protocol as described in Example 6 above were carried out in 8- to 12-week-old male db / db mice (C57BL / KsJ-db / db, weighing 35-45 g / mouse, supplied by Changzhou Cvens Experimental Animal Co. Ltd.).

[0240] An EMS skin biopsy punch with a diameter of 18 mm was used to create the wound.

[0241] Different drugs were administered topically at 50 μL per wound once daily from day 0 to day 12. The model group received the same amount of saline. There were seven groups containing 52 mice in this experiment, as shown in Table 4 below. [Table 4]

[0242] Each wound was photographed every other day starting on day 0. The photographs were scanned into a computer, and the wound area was calculated using ImageJ image analysis software (National Institutes of Health, USA).

[0243] The non-healed wound area was expressed as a percentage of the original wound area. A t / A0×100%, In the formula, A0 and A t refer to the initial area on day 0 and the wound area on the measurement day (time t), respectively.

[0244] The rate of unhealed wounds is shown in Figure 4. The results showed that Compound A was the most effective in improving wound healing and was superior to the combination of Compound 1 and montelukast.

[0245] The tissue specimens were analyzed, and skin regeneration, fibroblast proliferation, collagen regeneration score (Masson score), and inflammation score were estimated as follows:

[0246] The HE and Masson-stained sections were observed under a light microscope and scored (1, 2, or 3 points) according to the following criteria: skin regeneration score was 1 point if the area covered by newly generated skin was less than one-third of the wound area, 2 points if the newly generated skin covered more than one-third but less than two-thirds of the wound area, and 3 points if the area covered by newly generated skin was at least two-thirds of the wound area.

[0247] The skin regeneration scores are shown in FIG.

[0248] Fibroblast proliferation was scored according to the following criteria and is presented in Figure 6. [Table 5]

[0249] The results of the pathological analysis showed that Compound A and Compound 1 could promote skin regeneration and fibroblast proliferation. The conjugate Compound A was slightly superior, especially in terms of fibroblast proliferation score.

[0250] Example 8 Diabetic Wound Model II Similar experiments using essentially the same protocol as described in Example 7 above were performed on 8- to 12-week-old male db / db mice (C57BL / KsJ-db / db, weighing 35-45 g / mouse, supplied by Changzhou Cvens Experimental Animal Co. Ltd.).

[0251] Different concentrations of Compound A and Compound 1 ("A" and "1", respectively, as shown in Table 5 below) were prepared in substantially the same manner as described in Examples 6 and 7 above. Medium- and low-dose montelukast sodium ("Mon" in Table 5 below, MedChemExpress, MCE China, Shanghai, China) were dissolved in ultrapure water to obtain solutions with the concentrations listed in Table 5 below (L, M, and H indicate low, medium, and high doses, respectively). Considering the low solubility of montelukast in water, high-dose montelukast test samples were prepared by dissolving montelukast in 100% ethanol and then adding ultrapure water to form a solution with a concentration of 20 μg / μL in 20% ethanol.

[0252] As shown in Table 5 below, different drugs were administered topically at 50 μL / wound once daily from day 0 to day 12. The control group did not receive a wound. [Table 6]

[0253] The model group received the same amount of saline. There were eight mice in each group. Four mice were included in the control group. Skin pieces collected during wound creation were used as samples on day 7 for the control group.

[0254] The effect of the drugs on wound healing during the first 12 days is shown in Table 6 and Figure 7, which show the percentage of the remaining wound area of ​​the initial wound in the different groups (±standard deviation in the case of Table 6). [Table 7]

[0255] The data show that low doses of Compound 1 promoted wound healing in the early stages after wound injury, while medium doses of Compound A promoted better effects in the later stages.

[0256] Histological specimens were analyzed as described in Example 7 above, except that the collagen deposition scoring criteria for Masson-stained samples were as follows: no clear blue staining was given 0 points, blue fibers appearing in a dispersed pattern were scored as 1 point, if more blue fibers appeared this was scored as 2 points, and diffuse blue was given 3 points.

[0257] The results of histopathological analysis are shown in Figures 8 to 11 and indicate that all treatment groups promoted wound healing, and that the medium dose of Compound A in particular showed a significant promoting effect on collagen deposition.

[0258] The level of edema in the different groups was also assessed by histopathological analysis and the results are shown in Figure 12. The data show that the medium dose of Compound A produced the best results.

[0259] Example 9 Mouse ear swelling model II A similar experiment using essentially the same protocol as described in Example 5 above was conducted on 35 healthy male BALB / c mice. The mice were randomly divided into seven groups with five mice in each group, as described in Table 7 below. Compounds A, B, C, and D were all obtained from GL Biochem Ltd.

[0260] In Table 7 below, hydrogels of Compounds A, B, C, and D were prepared consisting of the listed amounts of active ingredient along with methylcellulose (2.5%), propanediol (11%), glycerol (11%), and acetic acid (pH adjuster, 0-0.5 g). All excipients were obtained from Sinopharm Chemical Reagent Co. Ltd. The gels were made up with water for injection.

[0261] Dexamethasone acetate cream (5 mg DEX in 10 g cream, Fuyuan Pharmaceutical Co. Ltd., Anhui Province, China) was used as a positive control.

[0262] Next, 40 μL of various therapeutic agents was applied to the right ear of each group. [Table 8]

[0263] The results obtained are shown in Figure 13. All the conjugates had a very good effect on eliminating the edema caused by acute inflammation.

[0264] Example 10 Acute Wound Model II Similar experiments using essentially the same protocol as described in Example 6 above were performed in 6-8 week old male C57BL / 6 mice. Two circular wounds were created in the midline of the back using a 12 mm diameter EMS skin biopsy punch. The two circles were tangential to each other, and the skin between the circles was cut along the upper and lower tangent lines. The wounds were trimmed using scissors. The wounds were oval in shape.

[0265] Different drugs were administered topically at 50 μL per wound once daily from day 0 to day 7. The model group received the same amount of saline. There were 10 groups containing 80 mice in this experiment, as shown in Table 8 below. [Table 9]

[0266] As described in Example 6 above, photographs were taken of each wound every other day from day 0 to depict the unhealed wound area.

[0267] The rate of unhealed wounds is shown in Figure 14. The results show that all four conjugates (A, B, C, and D) showed comparable effects in promoting wound healing compared to the other groups.

[0268] Example 11 Cream formulation Sorbitan stearate (0.6 g), polysorbate-80 (1 g), hexadecanol (2 g), octanoic / decanoic acid glyceride (5 g), liquid paraffin (4 g), monostearic acid glyceride (2 g), and petrolatum (5 g) (all from Sinopharm Chemical Reagent Co. Ltd.) were mixed, stirred, and heated to 85°C until the mixture was completely dissolved.

[0269] Methylcellulose (0.5 g), glycerin (4 g), trehalose (0.5 g), polyethylene glycol 200 (4 g), phenoxyethanol (0.3 g), and ethylhexylglycerol (0.1 g) (all from Sinopharm Chemical Reagent Co. Ltd.) were mixed with purified water (69.45 g), stirred, and heated to 85°C to obtain a uniform colloidal suspension.

[0270] The two mixtures obtained above were mixed with silicone oil (0.5 g) using an emulsifier under rapid stirring for 5 minutes, and the resulting emulsion was cooled to 55°C.

[0271] Compound A (50 mg, see Example 1 above) was dissolved in purified water (1 g) and mixed with the emulsion mixture with stirring until homogeneous. The resulting mixture was allowed to cool to room temperature to obtain the final product.

[0272] Example 12 Spray Formulation I Hydroxypropylmethylcellulose (HPMC, 0.1 g), hydroxyethylcellulose (0.1 g), glucose (5 g), phenoxyalcohol (0.5 g) (all from Sinopharm Chemical Reagent Co., Ltd.), and purified water (93.25 g) were stirred together and heated to 85°C to obtain a homogeneous colloidal suspension. The mixture was then cooled to room temperature.

[0273] Compound A (50 mg, see Example 1 above) was dissolved in 1 g of purified water. This solution was added to the colloid mixture. After uniform mixing, the final product was obtained.

[0274] Example 13 Spray Formulation II A second spray was prepared using substantially the same procedure as described in Example 12 above by adding the same aqueous solution of Compound A, along with the same amounts of other ingredients and 94.05 g of purified water, to a colloidal mixture made from slightly more HPMC and hydroxyethyl cellulose (0.2 g each).

[0275] Example 14 Gel Formulation I This formulation was obtained using substantially the same procedure as described in Examples 12 and 13 above, by adding the same aqueous solution of Compound A, along with the same amounts of other ingredients and 91.45 g of purified water, to a colloidal mixture made from HPMC and hydroxyethyl cellulose (1 g each).

[0276] Example 15 Gel Formulation II A second gel was obtained using substantially the same procedure as described in Examples 12-14 above by adding the same aqueous solution of Compound A to a colloidal mixture made from 0.5 g of HPMC and 1.5 g of hydroxyethyl cellulose, along with the same amounts of other ingredients and 91.45 g of purified water.

[0277] Example 16 Gel Formulation III A third gel was obtained using essentially the same procedure as described in Examples 12-15 above. Methylcellulose (2.2 g), propanediol (11 g, both from Sinopharm Chemical Reagent Co., Ltd.), and glycerol (11 g) were first mixed with purified water (74.75 g). The same aqueous solution of Compound A was added to the resulting colloidal mixture to obtain the final product.

[0278] Example 17 Clinical Case I - Patient with Allergic Rhinitis A 45-year-old female patient with allergic rhinitis presented with periodic runny nose and nasal congestion.

[0279] Spray Formulation I (see Example 12 above) packaged in a nasal spray bottle was administered to each nostril separately, two to three times daily for five days.

[0280] Patients were instructed to discontinue use of existing medications (oral montelukast sodium and budesonide) from the first dose of the spray formulation.

[0281] The runny nose and congestion were significantly relieved by the second dose. The patient found that he no longer felt the need to take oral montelukast sodium while on the new formulation. Budesonide was found to lose its effectiveness within several months of use.

[0282] Example 18 Clinical Case II - Relief of Burn Patient Symptoms A male patient was recovering from a severe second-degree burn (VAS score 4-5) and was experiencing severe itching on the inside of his upper arm.

[0283] Spray Formulation I (see Example 12 above) was applied to the wound and the itching was relieved within 1 minute.

[0284] Example 19 Clinical Case III - Relief of symptoms in injured patients The patient underwent surgery and subsequently experienced severe pain from the surgical incision.

[0285] Spray Formulation I (see Example 12 above) was administered to the incision and pain was relieved within 1 minute.

[0286] Example 20 Clinical Case IV - Patient with Allergic Rhinitis Thirty-eight subjects with seasonal and / or persistent allergic rhinitis were enrolled in this study. The majority of subjects had suffered from the condition for years and had attempted treatment with several medications, including steroids. Subjects were instructed to discontinue use of their existing medications after the first dose of the spray formulation.

[0287] Spray Formulation I (see Example 12 above) packaged in a nasal spray bottle was administered to each nostril separately, twice daily for 7 days.

[0288] Four subjects did not complete the study. Feedback collected from the remaining 34 subjects is presented below in Table 9.

[0289] The symptom incidence rate is equal to the number of subjects with a particular symptom divided by the total number of subjects. The efficacy rate is equal to the number of subjects whose symptoms were alleviated divided by the total number of subjects with a particular symptom. [Table 10]

[0290] Patients found the spray formulation easy to administer and did not cause irritation. Nasal congestion was immediately relieved, followed by sneezing and itchy eyes. Sixteen of the subjects were checked by a clinician after using the spray for seven days. Fifty percent of patients had less turbinate swelling, 68.75% had less nasal secretions, and 43% had reduced mucosal edema. No side effects were reported.

[0291] Example 21 Clinical Case V - Sore throat relief by nebulization An 80-year-old Caucasian male experienced sensations associated with the onset of a cold. Symptoms included an itchy and sore throat and nasal congestion. Five milliliters of spray formulation I (see Example 12 above) was loaded into a portable nebulizer (Feellife Medical Inc., Shenzhen, China). The suction nozzle of the nebulizer was placed in the mouth, and the device was turned on. Treatment lasted approximately 10 minutes. Only one inhalation was administered. The next morning, all cold symptoms had disappeared.

[0292] Example 22 Clinical Case VI - Relief of surgical pain in burn patients A patient with large, deep second-degree burns covering his entire back was admitted to the burn department of Beijing Shishuitan Hospital. He was treated for severe burns and suffered excruciating surgical pain with a VAS score of 7-9 every time a bandage was changed.

[0293] Spray formulation I (see Example 12 above) packed in a spray bottle was sprayed directly onto the burn surface. After 5 minutes, the dressing was removed and replaced with a new one. According to the clinician's assessment, after using the spray, the pain of the operation was reduced by about two-thirds.

[0294] Example 23 Clinical Case VII - Relief of surgical pain in laser surgery patients The study examined two subjects who underwent skin pigmentation removal surgery using fractional laser treatment.

[0295] Spray formulation I (see Example 12 above) packed in a spray bottle was sprayed onto the surface of the surgical area. After 10 minutes, the laser surgery began. According to the clinician's assessment, after using the spray, the pain of the surgery was reduced by about one-third.

[0296] It is common for subjects who undergo such laser surgery to experience burning pain for about 30 minutes after treatment, however, in this study, the subjects did not experience any burning pain afterwards.

[0297] Example 24 Clinical Case VIII - Relief of fever and cough A 5-year-old boy caught a cold and developed a severe cough. His temperature rose to 38°C overnight and he complained of a sore throat.

[0298] Spray Formulation I (see Example 12 above) was bottled and administered as an oral spray four times daily. The following day, the fever and sore throat symptoms disappeared. The cough disappeared after three days.

[0299] Example 25 Clinical Case IX - Relief of Contact Dermatitis A 53-year-old woman presented with contact dermatitis on her neck, which caused rash and itching when she wore a metal necklace.

[0300] Spray formulation I (see Example 12 above) packed in a spray bottle was sprayed onto the affected area. The itching sensation was relieved within 5 minutes. After two doses (once in the evening and once the following morning), all symptoms disappeared.

[0301] Example 26 Clinical Case X - Relief of colds The patients were a 42-year-old woman and her 10-year-old son. They both had a cold and were suffering from a sore throat and runny nose.

[0302] Spray Formulation I (see Example 12 above) packaged in a spray bottle was administered as an oral spray. After two doses (one in the evening and one the following morning), all symptoms disappeared.

[0303] Example 27 Clinical Case XI - Allergic Skin Disorders A 27-year-old woman with sensitive skin presented with an acne-like allergic skin disorder accompanied by slight itching on her face. She also had patches of redness and swelling on her face.

[0304] Spray formulation I (see Example 12 above) was applied in a spray bottle directly to the affected area of ​​the face, two sprays at a time, three times daily. The itching sensation was relieved within 30 minutes. The lesions completely disappeared after two weeks.

[0305] Example 28 Animal Model I - Idiopathic Pulmonary Fibrosis (IPF) Experimental animals and group assignment: Seventy-two adult male Sprague Dawley rats were divided into six groups after 7 days of adaptive feeding: a sham-operated (no infection or treatment) group, an IPF model group (no treatment), a high-drug dose test group, a medium-drug dose test group, a low-drug dose test group, and a positive control group.

[0306] The dosage of Compound A (Example 1) was set at 0.5 mg / mL, 0.1 mg / mL, and 0.02 mg / mL as the high, medium, and low doses, respectively. Oral administration of pirfenidone (Etuary®, Beijing Continent Pharmaceutical Co., Ltd., Beijing, China) at a single bolus dose of 30 mg / kg served as a positive control drug.

[0307] Modeling and Administration: A pulmonary fibrosis model was established by intratracheal instillation of bleomycin. Rats were anesthetized and placed supine on the operating table to expose the trachea. Bleomycin (5 mg / kg) in saline was injected into the trachea through the gap between the tracheal cartilage rings. The sham-operated group received an equal volume of saline. Immediately after administration, the rats were lifted vertically and rotated to ensure uniform distribution of the drug. After approximately 5 days, the rats recovered, and then different drugs were administered to the rats for 28 consecutive days according to the model design. The experimental design is shown in Table 10 below. [Table 11]

[0308] The following observational indicators were investigated: 1) Daily general observations of the rats' activity, sensitivity to external stimuli, fur gloss, hair color, mouth, lips, nose, weight, diet, respiration, and mortality. 2) Determination of the lung organ coefficient and lung wet / dry weight ratio of rats (i.e., the ratio of the animal's lung weight organ to the animal's body weight, i.e., the ratio of internal organs to body weight). 3) During the formation of pulmonary fibrosis, the expression of growth factors (TGF-β), tumor necrosis factor-α (TNF-α), and other cytokines involved in the development of fibrosis were measured. Standard ELISA methods were used to detect the contents of TGF-β, TNF-α, IL-1β, malondialdehyde (MDA), and superoxide dismutase (SOD) activity in lung tissue. 4) Detection of the content of collagen and fibrin metabolites (hydroxyproline) in the lungs as a specific indicator for assessing the degree of pulmonary fibrosis. 5) Detection of histopathological changes in lung tissue, which is the most important and objective indicator for evaluating pulmonary fibrosis. The results show that Compound A inhibits the overproduction of TGF-β and inflammatory cytokines. The results also show that the test drug has antioxidant effects by increasing SOD production and reducing lipid oxidation.

[0309] Example 29 Animal Model II - Antitussive Experiments: Ammonia-Induced Cough Method in Mice Sixty mice were randomly divided into five groups according to body weight: a CMC-Na negative control group, a dextromethorphan hydrobromide positive control group, and high, medium, and low dose (of Compound A, Example 1) groups. Each group contained 12 mice, 6 males and 6 females.

[0310] The test drug was administered by nebulization (0.15 mL / min) for 1 minute once daily for 5 days. The positive control drug was administered by intragastric administration at 10 mg / kg once daily for 5 days.

[0311] One, two, and four hours after the last drug administration (either Compound A or a positive control drug), mice were placed in an inverted beaker. One mL of ammonia water (25.0–28.0%) was placed in a boiling water bath, allowed to evaporate, and then placed in the beaker. Mice were stimulated with ammonia vapor for a predetermined time: 63.1, 50.1, 39.8, 31.6, 25.1, 20.0, 15.9, or 12.6 seconds. The logarithmic difference between two adjacent stimulation times was set to 0.1, followed by 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, and 1.1. Next, mice were immediately transferred to a bell jar. A stethoscope was used to detect the number of coughs within one minute. A typical cough occurring three or more times per minute was referred to as a "cough," while a cough occurring less than three times per minute was referred to as a "no cough."

[0312] The ammonia stimulation time for the next mouse was determined according to the sequential principle: if the first mouse was coughing, the next mouse was stimulated for a shorter time, whereas if the first mouse was not coughing, the next mouse was stimulated for a longer time.

[0313] EDT 50 was defined as the ammonia challenge time at which half of the mice developed a "cough" as calculated by the following formula: EDT 50 =lg -1 c / n (C is equal to the sum of r and x, where r is the number of animals in each stimulation time group, x is the logarithm of the stimulation time, and n is the number of animals in each group.) The results are shown in Table 11 below. [Table 12]

[0314] The results show that Compound A has a positive effect on relieving cough.

[0315] Example 30 Animal model III - Expectoration experiments - Phenol red excretion method in mice. Fifty mice were randomly divided into five groups based on their body weight: normal saline, a negative control group, an ammonium chloride positive control group, and high, medium, and low doses of the test drug (Compound A, above, or Compound E, below). Each group contained 10 mice, five males and five females.

[0316] Compound A was administered by nebulization (0.15 mL / min) for 1 minute once a day for 5 days, and a positive control drug was administered by intragastric administration for 5 days.

[0317] Thirty minutes after the last dose of Compound A, 5% phenol red solution was injected intraperitoneally. After another 30 minutes, the mice were sacrificed. The neck skin was removed, and the trachea from the thyroid cartilage to the bifurcation was isolated and immersed in 5% sodium bicarbonate solution with constant shaking. The sodium bicarbonate solution was used to detect the phenol red content.

[0318] The absorbance at 558 nm was detected by spectrophotometry (721G spectrophotometer, Shanghai Jingke, Shanghai, China). The optical density values ​​were used to calculate the phenol red content in the trachea by reference to a phenol red standard curve. The results of each group and the negative control group were tested for significance by t-test. The results of the expectoration experiment are shown in Table 12 below. [Table 13]

[0319] The results show that Compound A reduces mucus production and therefore reduces phlegm.

[0320] Example 31 Effect of Compound A on the activity of human herpes simplex virus type II (HSV-II) Serum-free 1640 medium was prepared using RPMI1640 powder (1000 mL dosage, Thermo Fisher Scientific China), L-glutamine (0.29 g, Sinopharm Chemical Reagent Co. Ltd, Shanghai, China), sodium bicarbonate (2.2 g, Sinopharm Chemical Reagent Co.), HEPE (2.39 g, Thermo Fisher Scientific China), and deionized water (1000 mL).

[0321] The reagents were mixed until dissolved, and the solution was sterilized by filtration. The mixture was formulated as a complete medium containing 10% serum by adding 10% newborn bovine serum before use, or the mixture was formulated as a maintenance solution by adding 2% newborn bovine serum.

[0322] 20 mg of Compound A (Example 1) was dissolved in 1 ml of 0.9% aqueous sodium chloride solution to prepare a 20 μg / μL stock solution. 0.05 mL of the stock solution was added to 1.95 mL of complete (10%) medium to formulate a 500 μg / mL drug solution. In the following antiviral tests 3 and 4, the maintenance solution (2%) was used instead of the complete medium.

[0323] Working solutions with concentrations of 250, 125, 62.5, 31.25, 15.625, 7.8125, 3.9063, 1.9531, and 0.9766 μg / mL were prepared by two-fold dilution.

[0324] 20.34 mg of sodium lauryl sulfonate (SDS, manufactured by AMRESCO LLC, Solon, Ohio, USA, packaged by Biosharp Company, Hefei, China, purity: 99%) was dissolved in 10.17 mL of complete medium to produce a 2000 μg / mL stock solution. A similar stock solution was also prepared in the same manner using the maintenance solution from the antiviral test. Next, working solutions of the concentrations listed above were prepared by two-fold dilution.

[0325] 2.25 mg of acyclovir (ACV, Zhiyuan Pharmaceutical Co., Ltd., Wuxi, China, purity: 99.3%) was dissolved in 2.25 mL of complete medium to form a 1000 μg / mL stock solution. Similar stock solutions were also prepared in the same manner using the maintenance solution from the antiviral test. 0.8 mL of each stock solution was diluted two-fold to provide working solutions with concentrations of 500, 250, and 125 μg / mL. 0.2 mL of each stock solution was added to 1.95 mL of complete medium to provide a concentration of 100 μg / mL, which was then diluted to provide solutions with concentrations of 50, 25, and 12.5 μg / mL.

[0326] 1.HSV-2 virus toxicity test A 0.5 mL suspension of human herpes simplex virus type II (HSV-2, SAV strain, Shanghai Institute of Cell Biology) was inoculated onto a monolayer culture of Vero cells (Shanghai Institute of Cell Biology), and after 1 hour of adsorption, the virus suspension was removed.

[0327] Maintenance solution was added and the cells were cultured at 37°C and 5% CO2 until over 95% of the cells showed clear pathological changes under a microscope (Nikon ECLIPSE TS100 inverted phase-controlled microscope with imaging system). The cells were harvested and subjected to three freeze-thaw cycles, followed by centrifugation at 3000 rpm for 10 minutes in a Model 400C Medical Low-Speed ​​Centrifuge (Beijing Baiyang Centrifuge Co., Ltd.). The supernatant was collected as the virus solution.

[0328] 2×10 5 A Vero cell suspension with a density of (cell number) was inoculated into a 96-well culture plate (Costar, Corning Inc., Oneonta, NY, USA) at 0.1 mL / well and cultured at 37°C under 5% CO2 in a Thermo Scientific CO2 incubator for 18 hours until a monolayer was visible under a microscope. The collected virus was diluted 10-fold with 0.1 mL / well of maintenance solution and inoculated onto the Vero cell monolayer. The maintenance solution was replenished and the cells were cultured at 37°C under 5% CO2. After 24 hours of culture, the cells were observed for pathological changes under a microscope. Each dilution was repeated in three wells. Normal cells served as an experimental control. The virus pathogenicity test was repeated three times.

[0329] Three fields were observed for each well, and the average percentage of pathological cells (P) within the field was determined.

[0330] Median viral infectious dose (TCID 50 , 50% of the tissue culture infectious dose of the virus, is determined by the traditional method of Reed and Muench, i.e., TCID 50 The logarithm of the dilution that gives a mortality rate above 50% is calculated as follows: logarithm of the dilution that gives a mortality rate above 50% - (logarithm difference x logarithm of the dilution factor). Generally, the following formula is used to calculate the "logarithm difference" (logarithm difference is also known as the "proportional distance" or "interpolated value"): logarithm difference = [(mortality rate at dilution above 50%) - 50%] / [(next mortality rate above 50%) - (next mortality rate below 50%)].

[0331] 2. Cytotoxicity of Compound A and control drugs Vero cells were seeded into 96-well culture plates and grown to monolayers. 0.2 mL of either Compound A solution (Example 1) or a control drug (20.34 mg sodium lauryl sulfonate or 2.25 mg acyclovir, as described above) was added to each well containing the correct (as described above) different concentrations of complete medium. This was repeated for each concentration in triplicate wells.

[0332] Solvent and normal cell culture were used as negative controls. Cells were cultured at 37°C and 5% CO2, and cell growth and morphological changes were observed under a microscope for 2 days. Three fields were selected for each well under a microscope, and the percentage of pathological cells was counted and the average value was calculated. The evaluation time point for the test was set at 24 hours, and the median toxic concentration (TC 50 ) and the maximum non-toxic concentration (TC0) were calculated. The experiment was repeated three times.

[0333] Cells were seeded as described above. Vehicle and normal cell culture were used as negative controls. 24 hours after adding Compound A or control drugs, 5 mg / mL 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT, Sigma-Aldrich, Shanghai, China) in PBS (diluted from a 10x stock solution, Sigma-Aldrich, China) was added (20 μL / well), and the culture was continued for 4 hours. The supernatant was then discarded from each well, and 150 μL of dimethyl sulfoxide (DMSO, Sigma-Aldrich, China) was added. The wells were then shaken in the dark at room temperature for 10 minutes.

[0334] Optical absorption value at 550 nm (OD 550 ) was measured by enzyme-linked immunosorbent meter (Multiskan Spectrum, Thermo Scientific, Shanghai, China).

[0335] 3. Effect of test drugs and SDS on the cytopathic effect of HSV-2 after direct action on the virus Determined TCID 50 HSV-2 virus stored at -80°C (in a Haier DW-86L486 ultra-low temperature freezer) was determined to have a titer of 200 TCID 50 (TCID 50 The value was initially determined for each time point and was 200 TCID 50 200 TCID 50The solution had a viral titer of 100 TCID 50 The mixture was mixed with an equal volume of either Compound A or SDS solution. The mixture was incubated at 37°C for 1 hour in a water bath (DK-8B thermostatically heated water bath, Shanghai Jinghong Biotech Co., Ltd.) and then inoculated into a 96-well culture plate containing a monolayer of Vero cells. 0.1 mL of the mixture was added to each well.

[0336] The supernatant containing the virus and drug was discarded after 1 hour of adsorption. Next, the monolayer Vero cells were washed twice with maintenance solution. Finally, 0.2 mL of maintenance solution was added to each well. The resulting mixture was continuously incubated at 37°C under 5% CO2 until the cytopathic rate of the drug-free culture reached 95% under a microscope. The evaluation time point of the test was set at 24 hours.

[0337] In addition to the experimental groups, three control groups were tested in parallel: a solvent, a no-drug control (virus control), and a normal cell control. Each group consisted of three wells, and the experiment was repeated four times.

[0338] Virus cultures were incubated at 0.1, 1, 10, 100, and 1000 TCID 50 The cytopathic effect was observed at 100 TCID 50 0.1 TCID 50 There should be no cytopathic effect, otherwise the neutralization test would not have been established.

[0339] The evaluation criteria were the same as those in the viral toxicity test. Three fields were observed under a microscope for each well. The average percentage of pathological cells (P) within the field was determined, and the median infectious dose of the virus (TCID 50 ) was calculated according to the method of Reed and Muencl (described above).

[0340] Drug toxicity to Vero cells was determined by cell morphology, and antiviral tests were performed at non-toxic concentrations. After 1 hour of incubation with different concentrations of test drugs and SDS, 100 TCID 50 HSV-2 (SAV strain) was inoculated into monolayer Vero cell cultures.

[0341] The results show that the cytopathic effect of cells caused by viral infection was inhibited to various degrees, suggesting that compound A has an inhibitory effect against HSV-2.

[0342] 4. Effect of test drugs and ACV on HSV-2 (direct method) 100 TCID of virus 50 The solution was diluted to 100 μg / well and inoculated into 0.1 mL of a monolayer Vero cell culture in each well. After 1 hour of adsorption, the supernatant was discarded and the culture was washed twice with maintenance solution. Then, 0.2 mL / well of a solution of different concentrations of Compound A or a control drug (acyclovir) was added. The culture was continuously incubated at 37°C under 5% CO2. Each concentration was replicated three times.

[0343] In addition to the experimental groups, three control groups were tested in parallel: vehicle, no drug control (virus control), and normal cell control.

[0344] During the incubation period, pathological changes were observed under a microscope, and the test was terminated when the cytopathic rate of the virus control reached >95%. The test evaluation time point was 24 hours, and the experiment was repeated three times.

[0345] The criteria were the same as those used in the viral toxicity test, i.e., three fields were selected for microscopic examination in each well, the average percentage of pathological cells (P) in the field was measured, and the average of the three fields was taken.

[0346] The linear regression equation was calculated according to the percentage of cytopathic effect of each reagent concentration group against the drug concentration. IC 50 values ​​were calculated and significance tests of correlation coefficients were also calculated.

[0347] The results show that Compound A has antiviral effects.

[0348] Example 32 Synthesis of montelukast-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (i.e., montelukast covalently bound at the N-terminus to amino acid SEQ ID NO: 12) The procedure described in Example 1 was repeated. A second product peak was detected at 5.813 minutes by LCMS (analytical column model: GS-120-5-C18-BIO, 4.6*250mm, detection: UV at 220nm, solvent A: 0.1% TFA in MeCN, solvent A: 0.1% TFA in water, flow rate 1.0mL / min, volume: 10μL) and compound.

[0349] MS: m / z 875.90[M+2H] 2+ .

[0350] Based on the characterization data available and presented herein, it is understood that the compound isolated in this example is that identified above as the title compound. The compound of Example 32 is hereinafter referred to as "Compound E."

[0351] The yield ratio of compound E to compound A was 1:9.

[0352] Example 33 Synthesis of hydrogenated montelukast styrene-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (i.e., hydrogenated montelukast styrene covalently bound at the N-terminus to amino acid SEQ ID NO: 12) The synthesis of the above compound was exactly the same as the procedure for Compound A described in Example 1, except that montelukast styrene was used as the reagent instead of montelukast.

[0353] MS: m / z 867.91[M+2H] 2+ .

[0354] Based on the characterization data available and presented herein, it is understood that the compound prepared in this example is the one identified above as the title compound. Otherwise, the compound prepared in Example 33 is a compound of the invention in which n is 0 in a compound of Formula I and the compound of Formula I is covalently linked at its N-terminus to amino acid sequence SEQ ID NO: 12. In any event, the compound of Example 33 is hereinafter referred to as "Compound F."

[0355] Example 34 Clinical Case XII - Relief of fever An 11-year-old boy presented with a fever of 39°C at 9:00 PM. The subject also had an intermittent cough and a runny nose.

[0356] A spray formulation of Compound E (2 mg, see Example 32 above) in normal saline (5 mL) was administered as a mist to each nostril over a 5-minute period at 22:00 by nebulization (device: handheld nebulizer, Lifetrons Beaute NS-400).

[0357] At 22:15, the subject fell asleep. Around midnight, the subject began to sweat and his body temperature dropped slightly. A second dose of Compound E (1 mg) in normal saline (2.5 mL) was administered in the same manner at 00:30.

[0358] By 03:30, the subject's temperature had dropped to 37.0°C. By 08:00, the subject was normal. Thus, between 22:15 the previous day and 08:00 the next day, only 30 minutes of coughing was observed in total. There was no observable nasal congestion during the night's sleep. In the morning, the runny nose returned, but was significantly improved compared to 11 hours earlier.

[0359] The subject received a third dose of Compound E (1 mg) in normal saline (2.5 mL) administered in the same manner at 8:30 AM that same morning. Two hours later, his runny nose stopped. The subject remained fever-free, cough-free, and runny nose-free until 3:00 PM that same day.

[0360] At 15:30 on the second day, the subject received a fourth dose of Compound E (1 mg) in normal saline (10 mL) by nebulization (device: Yuyue, air-compressed nebulizer, 403M). At 20:45, the subject's body temperature was 37.1°C. At 21:00, the subject received a fifth dose of Compound E (1 mg) in normal saline (10 mL) by nebulization. By 22:15, the subject's body temperature was 36.8°C.

[0361] Example 35 Comparison of Compounds A and E in a Mouse Ear Swelling Model (III) A similar experiment using essentially the same protocol as described in Example 5 above was conducted on 30 healthy male BALB / c mice. The mice were randomly divided into six groups with five mice in each group, as described in Table 13 below. [Table 14]

[0362] Compounds A and E were obtained from GL Biochem Ltd. and synthesized as described in Examples 1 and 32, respectively. Aqueous solutions of compounds A and E were prepared by dissolving 0.5 mg of powder in 1 mL of saline (0.9% w / v NaCl solution). 40 μL of the prepared solution was applied to the right ear of each group.

[0363] Dexamethasone acetate cream (5 mg DEX per 10 g of cream, Fuyuan Pharmaceutical Co. Ltd., Anhui, China), budesonide nasal spray (32 μg / spray × 120 sprays, 0.64 mg / ml, AstraZeneca AB, SE-151 85, Södertälje, Sweden), and fluticasone propionate nasal spray (50 mcg / spray, 0.05% w / w, Glaxo Wellcome, SA, Avenida de Extremadura no. 3-09400, Aranda de Duero, Burgos, Spain) were used as positive controls. The creams were placed in 1 mL syringes, and the dose was measured based on weight and volume calibration. The spray bottle was opened, and 40 μL of liquid was pipetted and applied to the right ear of each group.

[0364] The results obtained are shown in FIG.

[0365] All conjugates were highly effective in eliminating edema caused by acute inflammation. Compounds A and E had similar anti-inflammatory effects.

[0366] Example 36 Synthesis of montelukast-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys (i.e., montelukast covalently bound at the N-terminus to amino acid SEQ ID NO: 22) and montelukast styrene-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys (i.e., montelukast styrene covalently bound at the N-terminus to amino acid SEQ ID NO: 22) A modified peptide having SEQ ID NO:22 was synthesized using essentially the same procedure as described in Example 1 above, but with the order of coupling steps adjusted to provide for the amino acid sequence shown above.

[0367] Subsequently, montelukast was coupled to the N-terminus of Ala. The two title compounds were separated and purified by LCMS in the same manner as described in Example 32 above. Hereinafter, these compounds are referred to as Compound G (containing montelukast) and Compound H (containing montelukast styrene), respectively.

[0368] Yield ratio of compound G:H 1:7 MS (Compound G): m / z 876.6[M+2H]2 + MS (Compound H): m / z 867.6[M+2H]2 +

[0369] Example 37 In vitro CysLTR1 FLIPR antagonist testing The in vitro antagonist effects of Compounds A and E (see Examples 1 and 32 above, respectively) on the CYSLTR1 cell line were measured using a Fluo-4Direct™ calcium assay kit (catalog number F10471, Thermo Fisher Scientific). For comparison, montelukast sodium and montelukast styrene were also tested, and pranlukast was used as a positive control. Ten concentrations of each compound were tested in duplicate.

[0370] The CYSLTR1 / HEK293 cell line was used. Cells were prepared and 20 μL of the cell suspension was added to a 384-well plate (20K / well, poly-D-lysine protein-coated plate, Greiner #781946). The plate was placed in a 5% CO2 incubator at 37°C overnight.

[0371] Probenecid in FLIPR assay buffer was prepared from the relevant starter pack (catalog number F10471): A 250 mM stock solution of water-soluble probenecid was prepared by adding 1 mL of Fluo-4Direct™ calcium assay buffer to a 77 mg vial containing probenecid (component B of catalog number F10471). 10 mL of Fluo-4Direct™ calcium assay buffer and 200 μL of 250 mM probenecid stock solution were added to one bottle of Fluo-4Direct™ calcium reagent (component A). This 2× Fluo-4Direct™ calcium reagent loading solution was sufficient for two microplates. To completely dissolve the reagent, the solution was vortexed and allowed to stand for 5 minutes (protected from light). The reagent was prepared fresh daily.

[0372] All compounds were dissolved in Fluo-4Direct™ calcium assay buffer (without probenecid) and serially diluted. The cell plate was removed from the incubator, and the medium was gently decanted. 20 μL of compound was transferred to the cell plate, and 20 μL of 2× Fluo-4Direct™ no-wash loading buffer was added. The final concentrations of each compound were 100, 30, 10, 3, 1, 0.3, 0.1, and 0.03 μM. The plate was incubated at 37°C in a 5% CO2 incubator for 50 minutes and at room temperature for 10 minutes. Fluorescence was measured using appropriate instrument settings for excitation at 494 nm and emission at 516 nm.

[0373] The data was analyzed using Prism (GraphPad Software, USA) and the IC50 was calculated for each compound. The results are shown in Table 14 below. [Table 15]

[0374] The results show that montelukast has 11-fold greater affinity for CysLTR1 than montelukast styrene. Thus, compound E has 5-fold greater affinity than compound A. The affinity of the test compounds for CysLTR1 was primarily determined by the structure of the compound of formula I.

[0375] Example 38 In vitro CysLTR1 FLIPR antagonist testing The assay procedure described in Example 37 above was repeated for compound G (see Example 36 above), and for a further compound E (see Example 32 above), as a comparison. The results are shown in Table 15. [Table 16]

[0376] The results showed that compounds G and H had the same level of affinity for CysLTR1, indicating that changes in the amino acid sequence of the peptides had little effect on the affinity of the conjugates.

[0377] Example 38 Lipopolysaccharide-induced lung injury in mice Thirty-six BALB / c male mice weighing 20–22 g were housed in the animal facility at 22–26°C, 55–75% relative humidity, and a 12 / 12-h day / night cycle with food and water available ad libitum.

[0378] Mice were randomly divided into six groups as shown in Table 16 below. [Table 17]

[0379] Compound A was prepared similarly to the procedure described in Example 1 above, and Compound G was prepared similarly to the procedure described in Example 36 above.

[0380] Mice were anesthetized by intraperitoneal injection of 3% chloral hydrate (0.1 mL / 10 g). The tongue was pulled aside with forceps. Lipopolysaccharide (LPS, 1 mg / mL, 50 μL) was pipetted into the posterior pharyngeal wall. The tongue was then released, and the nose was immediately pinched for 30 seconds. The mice were then released from the restraint and returned to their cages to wake up naturally. Control mice were treated with an equal volume of saline.

[0381] Test compounds were administered by nebulization / inhalation 30 minutes after LPS challenge. Mice were sacrificed 24 hours later.

[0382] The thoracic cavity was immediately opened and the entire lung was removed. The lung tissue pieces were accurately weighed, and saline was added at a ratio of 9 mL per 1 g of lung tissue. The tissue was then homogenized and centrifuged at 3000 rpm for 10 minutes. The homogenate was used to detect TGF-β1 using an ELISA kit (Beijing 4A Biotech Co., Ltd., China), and the results are shown in Figure 16.

[0383] The results showed that both compounds of the present invention reduced inflammatory cytokines in lung tissue. The IL-1β concentration in lung tissue of Compound A group was at the same level as that of the low dose group of Compound G. Compound G also showed dose-dependent effectiveness in reducing inflammatory cytokines.

[0384] Thus, both compounds of the present invention have been shown to be effective in treating LPS-induced acute lung injury in mice, although the potency of Compound G was approximately 5-fold greater than that of Compound A.

[0385] Example 39 Idiopathic pulmonary fibrosis (IPF) model in rats Sixty adult SD rats (30 males and 30 females) were purchased from the Zhejiang Experimental Animal Center, China. The animals were housed at 21–26°C and 40–70% relative humidity with free access to food and water.

[0386] After 7 days of adaptation feeding, the rats were randomly divided into 6 groups as shown in Table 17 below. [Table 18]

[0387] The rats were anesthetized and placed supine on the operating table to expose the trachea. Bleomycin (5 mg / kg, injectable bleomycin hydrochloride, Haizheng Pfizer Pharmaceutical Co., Ltd.) and saline were injected into the trachea through the gap between the tracheal cartilage rings.

[0388] Sham-operated rats received an equal volume of saline instead of bleomycin. Immediately after administration, rats were lifted vertically and rotated to ensure uniform distribution of the bleomycin.

[0389] After about 7 days, rats recover, and then according to the model plan, rats are administered different drugs.6.5mg of the test compound of the present invention in powder form is accurately dissolved in 5mL of physiological saline to make a 1.3mg / mL solution.0.15mL of the solution is nebulized and each rat is allowed to inhale.Inhalation is carried out once a day.

[0390] In the pirfenidone group, 12 pirfenidone capsules (Beijing Contini Pharmaceutical Co., Ltd., Beijing, China, 100 mg) were opened and the contents were thoroughly suspended in 25 mL of 0.5% CMC-Na solution to obtain a 48 mg / mL suspension. The dose of pirfenidone was 1.0 mL / 200 g, i.e., 240 mg / kg, and administered orally by gavage to rats.

[0391] After 28 days of administration, the rats were anesthetized by intraperitoneal injection of chloral hydrate and sacrificed. The thoracic cavity was immediately opened and the entire lung tissue was removed. The wet lung weight was measured and the lung index was calculated (wet lung weight / rat weight x 1000), as shown in Table 18 below. [Table 19]

[0392] The results show that compounds A, E, and H reduce pulmonary edema caused by bleomycin induction.

[0393] The right bronchus was ligated, and the left lung was perfused in vitro with formalin solution. The left lung was dissected and fixed in formalin solution for pathological examination. The remaining tissue was stored in a -80°C refrigerator for later use.

[0394] Fixed lung tissues were embedded in paraffin, and serial 4-μm sections were stained with hematoxylin-eosin (HE) and modified Masson's trichrome stain. Fibrotic lung injury was morphologically evaluated using semiquantitative parameters. All morphological changes were scored according to the severity of the injury. Scores were given from 1 to 4, representing very mild, mild, moderate, and severe, respectively. No lesions were scored as 0. The score from HE-stained sections was the sum of the degrees of fibrosis and inflammation. The score from Masson-stained sections was the degree of collagen deposition in the lung interstitium. The results are shown in Table 19 below. [Table 20]

[0395] The results showed that pulmonary fibrosis and bronchopneumonia were more severe in the model group than in the sham-operated group. Compared with the model group, the pathological changes in the drug-treated groups were similar, but the degree of pathological changes was smaller. The order of pathological changes was as follows: model, pirfenidone > Compound E > Compound A > Compound H > sham. These results indicated that compounds A, E, and H could prevent bleomycin-induced pulmonary fibrosis in mice, and their effectiveness was stronger than that of pirfenidone.

[0396] Example 40 Synthesis of Montelukast-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (i.e., montelukast styrene covalently attached at the N-terminus to amino acid SEQ ID NO: 20) A modified peptide having SEQ ID NO:20 was synthesized using essentially the same procedure as described in Example 1 above, but with the order of coupling steps adjusted to provide for the amino acid sequence shown above.

[0397] Subsequently, montelukast was coupled to the N-terminus of Ala. The compound was purified by LCMS in a manner similar to that described in Example 32 above. Hereafter, the title compound is referred to as Compound J.

[0398] MS: m / z 924.15 [M+2H]2+.

[0399] Example 41 Mouse ear swelling model IV Essentially the same protocol as described in Example 5 above was carried out on 15 healthy male BALB / c mice using Compound J (see Example 40 above) as the test compound. The mice were randomly divided into three groups with five mice in each group, as described in Table 20 below.

[0400] Compound J hydrogel was prepared containing 0.5 mg / g of the active ingredient and methylcellulose (2.5%), propanediol (11%), glycerol (11%), and acetic acid (pH adjuster, 0-0.5 g). All excipients were obtained from Sinopharm Chemical Reagent Co., Ltd. The gel was made with water for injection.

[0401] Dexamethasone acetate cream (DEX cream, 5 mg dexamethasone in 10 g cream, Fuyuan Pharmaceutical Co. Ltd., Anhui Province, China) was used as a positive control. Next, 40 μL of various therapeutic agents were applied to the right ear of each group. [Table 21]

[0402] The results are shown in Figure 17. Compound J showed a very good effect in removing edema caused by acute inflammation.

[0403] Example 42 Clinical Cases - Allergic Conjunctivitis A 52-year-old female patient was diagnosed with allergic conjunctivitis and experienced eyelid swelling, itching, and a runny nose.

[0404] Compound G was dispensed in a spray bottle at 0.5 mg / mL in saline. The spray was administered to each eye two to three times daily for seven days.

[0405] The patient felt relief from itchy eyes after one treatment. On the second day of treatment, her eyelids were less swollen. Complete resolution of all symptoms occurred within one week.

[0406] Example 43 Clinical case - ulcerative colitis A hospitalized patient with ulcerative colitis suffered from severe symptoms, including severe abdominal pain and cramps, frequent diarrhea (more than 20 times per day), and was not responding to over-the-counter medications. The patient experienced rectal bleeding, passing small amounts of blood in the stool, urgency to defecate, and fever.

[0407] A hydrogel of Compound G was prepared, consisting of 0.5 mg / g of the active ingredient, methylcellulose (2.5%), propanediol (11%), glycerol (11%), and acetic acid (pH adjuster, 0–0.5 g). All excipients were obtained from Sinopharm Chemical Reagent Co. Ltd. The gel was made with water for injection.

[0408] The patient was given 2 g of the above gel by anal administration once daily. After 2 days of administration, diarrhea decreased to 5-6 times a day and bleeding became less frequent.

[0409] Patients continued to use the gel to determine its long-term effectiveness.

Claims

1. 1. A peptide-containing compound comprising a peptide component, the peptide component having the amino acid sequence: X-Pro-Y-Z (SEQ ID NO: 5) wherein X represents a chain of 1 to 2 amino acid residues independently selected from the group consisting of Ala and Lys; Y is selected from the group consisting of Ser and pSer; Z represents a chain of 1 to 7 amino acid residues independently selected from the group consisting of Tyr, pTyr, 3Hyp or 4Hyp, Thr, pThr, DOPA, and Lys; At least one of the Ala and / or Lys residues is linked to one or more compounds of formula I: 【Chemical 1】 During the ceremony, R １ は、-C(CH ３ ) ２ OH、-COCH ３ 、-C(CH ３ )=CH ２ 、 and -C(CH ３ ) ２ H; a peptide-containing compound, wherein n is 0, 1, or 2; Or a stereoisomer, or a pharmaceutically or cosmetically acceptable salt of said peptide-containing compound.

2. 2. The peptide-containing compound of claim 1, wherein the peptide component is coupled to at least one compound of Formula I via a primary amide linkage formed at the N-terminus of the peptide component.

3. The peptide component has at least one free —NH in one or more amino acids in the sequence that is not the N-terminal amino acid. ２ 3. The peptide-containing compound of claim 1 or 2, which is coupled to at least one compound of formula I via a primary amide linkage formed through a group.

4. 4. The peptide-containing compound of claim 1, wherein at least about 5% of the amino acids are lysine (Lys) and / or at least about 20% of the amino acids are lysine (Lys).

5. 5. The peptide-containing compound of claim 1, wherein at least about 5% of the amino acids of the peptide component contain an aromatic group.

6. The peptide-containing compound according to any one of claims 1 to 5, wherein the peptide component is an antibacterial and / or anti-inflammatory peptide.

7. 7. The peptide-containing compound of claim 1, wherein at least about 10% of the amino acids contain an aromatic group.

8. 8. The peptide-containing compound of claim 5 or 7, wherein the amino acid containing an aromatic group is selected from the group consisting of tyrosine (Tyr) and 3,4-dihydroxyphenylalanine (DOPA).

9. 9. The peptide-containing compound of claim 1, wherein the peptide component comprises one or more amino acid fragments / sequences of a mussel adhesive protein.

10. 10. The peptide-containing compound of claim 9, wherein the mussel adhesive protein is mefp-1.

11. The peptide component has the amino acid sequence: G １ -Lys-Pro-G ２ -T-Hyp-G ３ -Lys (arrangement number 7) comprising or consisting of: G １ represents absent or Ala, G ２ is selected from the group consisting of Ser and pSer; T is selected from the group consisting of DOPA, Tyr, and pTyr; Hyp is selected from the group consisting of 3Hyp and 4Hyp; G ３ represents a chain of 1 to 4 amino acid residues, each independently selected from the group consisting of Tyr, pTyr, 3Hyp, 4Hyp, Thr, pThr, and DOPA; 11. The peptide-containing compound of any one of claims 1 to 10, wherein at least one of the Lys residues and / or Ala residues, if present, is linked to at least one compound of formula I as defined in claim 1.

12. The peptide component has the amino acid sequence: Lys-Pro-G ２ -T-Hyp-G ３ -Lys (array number 8) wherein G of claim 11 ２ , T, Hyp, and G ３ 12. The peptide-containing compound of claim 11, wherein:

13. G １ The peptide-containing compound of claim 11 , wherein represents Ala.

14. G ３ The peptide-containing compound according to any one of claims 11 to 13, wherein represents Tyr or DOPA.

15. The peptide component has the amino acid sequence: Ala-Lys-Pro-G ２ -T-Hyp-Hyp-Thr-G ４ -Lys (array number 9) wherein G comprises or consists of ２ , T, and Hyp are as defined in claim 12; ４ 14. The peptide-containing compound of claim 11 or 13, wherein is selected from the group consisting of Tyr, pTyr, 3Hyp, 4Hyp, Thr, pThr, and DOPA.

16. G ２ The peptide-containing compound according to any one of claims 11 to 14, wherein represents Ser.

17. The peptide-containing compound according to any one of claims 11 to 15, wherein T represents Tyr.

18. G ４ The peptide-containing compound according to any one of claims 15 to 17, wherein represents Tyr or DOPA.

19. The peptide component has the amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (SEQ ID NO: 4), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-DOPA-Lys (SEQ ID NO: 11), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 12), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 13), Lys-Pro-Ser-Tyr-Hyp-DOPA-Lys (SEQ ID NO: 14), and 12. The peptide-containing compound of claim 11, comprising or consisting of Lys-Pro-Ser-pTyr-Hyp-DOPA-Lys (SEQ ID NO: 15).

20. The peptide component has the amino acid sequence:

20. The peptide-containing compound of claim 19, comprising or consisting of Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID NO: 12).

21. G ３ But, -V １ -Thr-Tyr-V ２ represents -, and V １ binds to Hyp, and V ２ is bound to Lys, and V １ and V ２ 18. The peptide-containing compound of any one of claims 11 to 13, 16 or 17, wherein independently represent none, one or two Hyp groups.

22. -V １ -Thr-Tyr-V ２ The peptide-containing compound of claim 21, wherein - represents -Hyp-Thr-Tyr-, -Hyp-Thr-Tyr-Hyp-, or -Thr-Tyr-Hyp-.

23. The peptide component has the amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 18), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 19), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 20), Ala-Lys-Pro-pSer-Tyr-Hyp-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 21), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 22), Ala-Lys-Pro-pSer-Tyr-Hyp-Thr-DOPA-Hyp-Lys (SEQ ID NO: 23), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 24), and 12. The peptide-containing compound of claim 11, comprising or consisting of Ala-Lys-Pro-pSer-Tyr-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 25).

24. The peptide component has the amino acid sequence:

24. The peptide-containing compound of claim 23, comprising or consisting of Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys (SEQ ID NO: 24).

25. 25. The peptide-containing compound of any one of claims 1 to 24, wherein one or more compounds of formula I are linked via an N-terminal Ala or Lys moiety and / or a C-terminal Lys moiety.

26. 26. The peptide-containing compound of any one of claims 1 to 25, wherein one or two compounds of formula I are linked to the peptide component.

27. 27. The peptide-containing compound of claim 26, wherein one compound of formula I is linked to the peptide moiety.

28. In one or more compounds of formula I above, R １ But -C(CH ３ ) ２ OH, -C(CH ３ ) = CH ２ , and -C(CH ３ ) ２ 28. The peptide-containing compound of any one of claims 1 to 27, wherein n is selected from the group consisting of H and / or n is 0.

29. 29. The peptide-containing compound of any one of claims 1 to 28, wherein the one or more compounds of formula I is montelukast, montelukast styrene, or hydrogenated montelukast styrene.

30. 30. The peptide-containing compound of any one of claims 1 to 29, wherein the one or more compounds of formula I is montelukast.

31. 25. The peptide-containing compound of claim 24, wherein the compound of Formula I is montelukast, and the montelukast is covalently attached to an N-terminal Ala residue.

32. 32. The peptide-containing compound according to any one of claims 1 to 31, or a pharmaceutically or cosmetically acceptable salt thereof, for use as a pharmaceutical.

33. 32. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 31, or a pharmaceutically or cosmetically acceptable salt thereof, and a pharmaceutically or cosmetically acceptable adjuvant, diluent, or carrier.

34. 34. The pharmaceutical formulation of claim 33, which is suitable, adapted and / or packaged and presented for topical administration, and wherein the pharmaceutically or cosmetically acceptable adjuvant, diluent or carrier is a topical adjuvant, diluent or carrier.

35. 35. The formulation of claim 33 or claim 34 in the form of a gel, spray, cream, ointment or dry powder.

36. 36. The pharmaceutical formulation of any one of claims 33 to 35, further comprising another anti-inflammatory agent.

37. A kit of parts comprising: (A) a pharmaceutical formulation according to any one of claims 33 to 35; (B) a pharmaceutical formulation comprising another anti-inflammatory agent mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier; A kit of parts, wherein the components (A) and (B) are each provided in a form suitable for administration in combination with the other.

38. Use of a compound according to any one of claims 1 to 31, or a pharmaceutically or cosmetically acceptable salt thereof, a formulation according to any one of claims 33 to 36, or a kit of parts according to claim 37, for the manufacture of a medicament for the treatment of inflammation, an inflammatory disorder, and / or a disorder characterised by inflammation.

39. 39. The use of claim 38, wherein the inflammatory disorder comprises inflammation of one or more of soft tissue, joints, nerves, vasculature, one or more internal organs, one or more mucosal surfaces, or skin.

40. 40. The use according to claim 38 or 39, wherein the inflammatory disorder involves the oral, nasal, ocular, vaginal, cervical and / or anorectal mucosa.

41. The use according to any one of claims 38 to 40, wherein the inflammatory disorder is an allergic or atopic condition.

42. 42. The use of claim 38 or 41, wherein the inflammatory disorder is selected from the group consisting of pharyngitis, periodontitis, gingivitis, xerophthalmia, dermatitis, hives, food allergies, early symptoms of skin cancer, erythematous skin lesions, pathological alopecia, pathological alopecia after skin grafting, erythema multiforme, folliculitis, and otitis externa.

43. The use according to any one of claims 38 to 41, wherein the inflammatory disorder is selected from the group consisting of cystic fibrosis, common interstitial pneumonia, allergic pneumonia, asbestosis, emphysema, cor pulmonale, and pulmonary embolism.

44. The use according to any one of claims 38 to 41, wherein the underlying physiological disorder is characterized by oxidation.

45. 45. The use of claim 44, wherein the peptide-containing compound increases superoxide dismutase production and / or reduces lipid oxidation.

46. 39. The use of claim 38, wherein the disorder comprises inflammation of one or more mucosal surfaces and is selected from oral mucositis, aphthous ulcers, otitis media, laryngitis, tracheitis, esophagitis, gastritis, enteritis and enterocolitis (including bacterial dysentery, chronic amebic dysentery, schistosomiasis, nonspecific ulcerative colitis, and regional enteritis), cervicitis and endocervicitis, endometritis, inflammation caused by inhalation injury, mucosal inflammation associated with cancer, and mucosal inflammation associated with infection.

47. 39. The use of claim 38, wherein the inflammatory disorder comprises inflammation of the heart, stomach, intestine, lung, liver, spleen, kidney, pancreas, bladder, ovary, prostate, pericarditis, myocarditis, endocarditis, pneumonia, hepatitis, splenitis, nephritis, pancreatitis, cystitis, oophoritis, prostatitis or gastric ulcer.

48. 48. The use of claim 47, wherein the disorder is selected from psoriasis, acne, eczema, dermatitis, pathological alopecia, seborrheic dermatitis, rhinitis, hemorrhoids, chronic obstructive pulmonary disease, and inflammatory bowel disease.

49. 49. The use of claim 48, wherein the disorder is chronic obstructive pulmonary disease, emphysema, or chronic bronchitis.

50. 49. The use of claim 48, wherein the inflammatory bowel disease is ulcerative colitis.

51. 48. The use according to any one of claims 38 to 47, wherein the disorder is asthma, allergic rhinitis, or allergic / atopic dermatitis.

52. 39. The use of claim 38, wherein the disorder comprises inflammation of the eye and surrounding area.

53. 53. The use of claim 52, wherein the disorder is conjunctivitis, keratitis, or optic neuritis.

54. 54. The use of claim 53, wherein the disorder is selected from allergic conjunctivitis, acute superficial keratitis, nummular keratitis, interstitial keratitis, disciform keratitis, neurotrophic keratitis, mucoplaque keratitis, herpes simplex keratitis, herpes zoster keratitis, bacterial keratitis, fungal keratitis, acanthamoeba keratitis, Onchocerca volvulus keratitis, superficial punctate keratitis, ulcerative keratitis, lagophthalmos keratitis, photokeratitis, and acute hyperemia during contact lens wear.

55. 39. The use of claim 38, wherein the disorder characterized by inflammation is or results in a wound or a burn.

56. 56. The use of claim 55, wherein the wound comprises an open wound, an acute wound, a chronic wound, a contusion, a chemical burn, a drug rash, a polymorphous light eruption, a chemical rash, a sunburn, chilblains or frostbite.

57. 56. The use of claim 55, wherein the wound comprises an abrasion, scratch, incision, laceration, skin puncture, laceration, contusion, scar, or blister.

58. 58. The use according to any one of claims 55 to 57, wherein the wound results from a skin wound comprising superficial damage to the epidermis and / or dermis; and / or a full thickness skin wound comprising damage below the epidermis and / or dermis.

59. 59. The use according to any one of claims 55 to 58, wherein the wound results from a physical injury to the internal or external surface of the membrane.

60. 60. The use of claim 59, wherein the physical injury comprises surgery or skin grafting after a burn.

61. 59. The use according to any one of claims 55 to 58, wherein the wound is caused by an underlying physiological disorder.

62. 62. The use of any one of claims 55 to 58 or 61, wherein the wound is a chronic wound.

63. 63. The use according to claim 62, wherein the disorder resulting in wounds is hemorrhoids or ulcerative colitis or Crohn's disease.

64. 63. The use according to claim 62, wherein the chronic wound is a skin or mucosal ulcer.

65. 65. The use of claim 64, wherein the ulcer is a skin ulcer that is difficult to heal.

66. 66. Use according to any one of claims 55 to 65, wherein the treatment is for the prevention of physiological responses resulting from the inflammation and / or wound healing process selected from pain (including pain, procedural and / or non-procedural pain) associated with the inflammation and / or wound, pruritus (itching) associated with the wound itself and the healing process, exudation of fluid from the wound, risk of infection, and / or scarring and melanin pigmentation.

67. 62. The use of claim 61, wherein the underlying physiological disorder is associated with melanin pigmentation.

68. 68. The use of claim 67, wherein the physiological disorder associated with melanin pigmentation is melasma, freckles, melanin deposits, butterfly rash, pigmentation, skin cancer with melanoma, or pigmentation caused by sun exposure or skin diseases such as acne.

69. Use of a compound according to any one of claims 1 to 31, or a pharmaceutically or cosmetically acceptable salt thereof, a formulation according to any one of claims 33 to 36, or a kit of parts according to claim 37, for the manufacture of a medicament for the treatment of a pulmonary and / or fibrotic condition.

70. The condition may be pulmonary fibrosis, renal fibrosis, hepatic fibrosis, silicosis, acute bronchitis, chronic bronchitis, tracheobronchitis, bronchial asthma, severe asthma, bronchiectasis, upper respiratory tract infections including colds and influenza, allergic airway inflammation, bacterial pneumonia, viral pneumonia, mycoplasmal pneumonia, reckettsia, radiation pneumonitis, pneumococcal (including staphylococcus, streptococcus, and gram-negative bacillus) pneumonia, pulmonary candidiasis (including aspergillosis, mucormycosis, histoplasmosis, actinomycosis, and nocardiosis), pulmonary mycoses, cryptococcosis, lung abscess, anaphylactic pneumonia (Leoffer's syndrome), 70. The use of claim 69, wherein the allergic disease is selected from the group consisting of pulmonary eosinophilia (eosinophilia), extrinsic allergic alveolitis, pulmonary eosinophilia (hypeosinophilia), obstructive emphysema, pulmonary edema, pulmonary tuberculosis, respiratory alkalosis (acidosis), acute lung injury, interstitial lung disease, empyema, pulmonary fibroma, and cor pulmonale.

71. 71. The use of claim 70, wherein the condition is selected from idiopathic pulmonary fibrosis or acute lung injury.

72. Use of a compound according to any one of claims 1 to 31, or a pharmaceutically or cosmetically acceptable salt thereof, a formulation according to any one of claims 33 to 36, or a kit of parts according to claim 37, for the manufacture of a medicament for the treatment of a viral infection.

73. The viral infection is selected from the group consisting of adenovirus, human papillomavirus, BK virus, JC virus, herpes simplex type 1 virus, herpes simplex type 2 virus, varicella zoster virus, Epstein-Barr virus, human cytomegalovirus, human herpes virus type 8, smallpox, hepatitis B virus, parvovirus B19, human astrovirus, norovirus, Norwalk virus, coxsackievirus, hepatitis A virus, poliovirus, rhinovirus, coronovirus, hepatitis C virus, yellow fever virus, dengue virus, ue 73. The use of claim 72, wherein the infection is caused by one or more viruses selected from: Streptococcus aureus, Tick-borne encephalitis virus, Human immunodeficiency virus, Rubella virus, Lassa virus, Hantavirus, Crimean-Congo hemorrhagic fever virus, Hantavirus, Ebola virus, Marburg virus, Ravn virus, Influenza virus, Measles virus, Mumps virus, Parainfluenza virus, Respiratory syncytial virus, Rhabdoviridae, Hepatitis E virus, Rotavirus, Orbivirus, Coltivirus, Bannavirus, and Hepatitis D virus.

74. 74. The use of claim 73, wherein the virus is selected from adenovirus, human papillomavirus, herpes simplex, type 1 virus, herpes simplex, type 2 virus, rhinovirus, coronavirus, including influenza A virus, influenza B virus, or influenza C virus, parainfluenza virus, and respiratory syncytial virus.

75. 75. The use of claim 74, wherein the virus can cause infection of the respiratory tract.

76. 74. The use of claim 73, wherein the virus is a human immunodeficiency virus.

77. 33. The compound for use according to claim 32, wherein the compound(s) or salts thereof are administered topically in the form of a topical formulation.

78. 78. The compound for use according to claim 77, wherein the condition is treated by direct topical administration to the skin.

79. 78. The compound for use according to claim 77, wherein the condition is treated by direct topical administration to a mucosal surface.

80. 80. The compound for use according to claim 32 or any one of claims 77-79, wherein the compound(s) are administered by oral, intravenous, intra-arterial, intravascular, perivascular, intramuscular, cutaneous, subcutaneous, transmucosal, sublingual, buccal, rectal, intravaginal, transdermal, nasal, pulmonary, tracheal, bronchial, intraperitoneal, anorectal, or other parenteral delivery.

81. 81. A compound for use according to claim 80, characterized in that the administration is to the lungs in the form of a spray, including a powder aerosol or an aqueous mist used for atomization.

82. 82. The compound for use according to claim 81, wherein the spray is a liquid, including a water (aerosol) spray, and the excipients may include one or more of viscosity modifiers, sugars, emulsifiers, buffers, alcohols, and preservatives.

83. 83. The compound for use according to claim 81 or 82, wherein administration is performed from an inhalation device selected from a pressurized metered dose inhaler, a dry powder inhaler, a soft mist inhaler, or a nebulizer.

84. 77. The use according to any one of claims 38 to 76, wherein the compound(s) or salts thereof are administered topically in the form of a topical formulation.

85. 85. The use of claim 84, wherein the condition is treated by direct topical administration to the skin.

86. 85. The use of claim 84, wherein the condition is treated by direct topical administration to a mucosal surface.

87. 87. The use of any one of claims 38-76 or claims 84-86, wherein the compound(s) are administered by oral, intravenous, intra-arterial, intravascular, perivascular, intramuscular, cutaneous, subcutaneous, transmucosal, sublingual, buccal, rectal, intravaginal, transdermal, nasal, pulmonary, tracheal, bronchial, intraperitoneal, anorectal, or other parenteral delivery.

88. 88. The use according to claim 87, characterized in that the administration is to the lungs in the form of a spray comprising a powder aerosol or an aqueous mist used for atomization.

89. 89. The use of claim 88, wherein the spray is a liquid comprising a water (aerosol) spray and excipients may comprise one or more of viscosity modifiers, sugars, emulsifiers, buffers, alcohols, and preservatives.

90. 90. The use of claim 88 or 89, wherein administration is performed from an inhalation device selected from a pressurized metered dose inhaler, a dry powder inhaler, a soft mist inhaler, or a nebulizer.

91. 32. A process for the preparation of a peptide-containing compound according to any one of claims 1 to 31, comprising reacting a compound of formula I according to any one of claims 1 or 28 to 30 with a peptide component according to any one of claims 1 to 24.

92. 37. A process for the preparation of a pharmaceutical formulation according to any one of claims 33 to 36, comprising associating a peptide-containing compound according to any one of claims 1 to 31 with one or more pharmaceutically acceptable adjuvants, diluents, or carriers.

93. 38. A process for the preparation of a kit of parts according to claim 37, comprising associating component (A) of said kit of parts with component (B) of said kit of parts.

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