Novel multifunctional oligopeptides

Abstract

A compound of formula I is provided, A-Q-B I wherein Q represents a structural fragment of formula II, wherein: the wavy line and m have the meaning given in the description, and wherein A and B have the meaning given in the description, but can represent a peptide component having the following amino acid sequence: [W-Lys-X 1 -Ser-U-X 2 -Y] n -W-Lys-X 1 -Ser-U-X 2 -Y- - - (SEQ ID No: 3) wherein the dotted line, n, W, X 1 , U, X 2 and Y have the meaning given in the description, which is useful in medicine, including as pharmaceutical excipients, adhesives and film-forming materials, and / or which is useful in the treatment of diseases characterized by inflammation, including wounds; burns and mucosal disorders, such as anorectal diseases, inflammatory bowel diseases, gynaecological diseases and dental diseases.

Description

Technical Field

[0001] This invention relates to novel peptides, the use of such peptides as pharmaceutical active ingredients or otherwise in human medicine, and pharmaceutical compositions comprising said peptides. Specifically, this invention relates to the use of these peptides and compositions in the treatment of a variety of conditions, including inflammation. Background Technology

[0002] Inflammation is typically characterized as a localized tissue response to the invasion of, for example, microorganisms, certain antigens, damaged cells, or physical and / or chemical agents. The inflammatory response is generally a protective mechanism used to destroy, dilute, or isolate harmful agents and damaged tissue, and to initiate tissue healing.

[0003] Inflammation can be caused by physical trauma, infection, some chronic diseases (e.g., psoriasis and autoimmune diseases such as rheumatoid arthritis), and / or chemical and / or physiological responses to external stimuli (e.g., as part of an allergic reaction). It can involve a complex series of events in which inflammatory mediators increase blood flow and dilate local blood vessels, leading to redness and heat, fluid exudation, often resulting in local swelling, white blood cell migration to the inflamed area, and pain.

[0004] Many conditions / disorders are characterized by and / or caused by abnormal, tissue-damaging inflammation. These conditions are typically characterized by the activation of immune defense mechanisms, resulting in a greater harm than benefit to the host, and are usually associated with varying degrees of tissue redness or congestion, 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, such as increased blood flow through local vasodilation, resulting in redness and heat; extravasation of white blood cells and plasma, often leading to local swelling; activation of sensory nerves (causing pain in some tissues); and loss of function. These inflammatory changes are triggered by a series of cellular and biochemical events involving cells such as neutrophils, monocytes, macrophages, and lymphocytes, as well as inflammatory mediators such as vasoactive amines, cytokines, complement factors, and reactive oxygen species.

[0006] Furthermore, inflammation plays a crucial role in the wound healing process. Therefore, wounds and burns can be classified as inflammation-related conditions. The conventional wisdom in this field is that anti-inflammatory drugs should not be applied directly to open wounds, as this would hinder wound healing.

[0007] Fibrosis is defined by the excessive accumulation of fibrous connective tissue (components of the extracellular matrix (ECM), such as collagen and fibronectin) in and around inflamed or damaged tissue. Although collagen deposition is typically a reversible part of wound healing, it can often evolve into a progressively irreversible fibrotic reaction if tissue damage is severe or if the wound healing response itself becomes disordered. Furthermore, fibrosis is known to be a leading cause of morbidity and death in many chronic inflammatory diseases, as well as end-stage liver disease, kidney disease, idiopathic pulmonary fibrosis (IPF), and heart failure. It is also a pathological feature of many chronic autoimmune diseases, such as scleroderma, rheumatoid arthritis, Crohn's disease, ulcerative colitis, myelofibrosis, and systemic lupus erythematosus. Fibrosis can also influence the pathogenesis of many progressive muscle diseases, metastases, and transplant rejection.

[0008] Mussel adhesion protein (MAP) (also known as mussel byssal protein (mefp)) is a protein secreted by marine mollusks such as the mussel (Mytilus edulis), the thick-shelled mussel (Mytilus coruscus), and the green mussel (Pernaviridis). Eleven independent adhesion protein subtypes have been identified from mussels, including collagen pre-COL-P, pre-COL-D, and pre-COL-NG; mussel byssal matrix proteins PTMP (proximal filament matrix protein) and DTMP (distal filament matrix protein); and mfp proteins mfp-2 (sometimes referred to as “mefp-2”, which can be used interchangeably below), mfp-3 / mefp-3, mfp-4 / mefp-4, mfp-5 / mefp-5, mfp-6 / mefp-6, and the most preferred mfp-1 / mefp-1 (see, for example, Zhu et al., Advances in Marine Science, 2014, 32, 560-568 and Gao et al., Journal of Anhui Agr. Sci., 2011, 39, 19860-19862).

[0009] A key portion of mefp-1 consists of 70 to 90 tandem decapeptide repeats of the following sequence: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys (SEQ ID No: 1; see Waite, Int. J. Adhesion and Adhesives, 1987, 7, 9-14). This decapeptide sequence can be isolated as a low molecular weight derivative of naturally occurring MAP, or it can be synthesized, for example, as described in J. Chem. Soc., Perkin Trans., 1987, 1, 613-618. See also Dalsin et al., J. Am. Chem. Soc., 2003, 125, 4253-4258.

[0010] Decapeptide analogs have also been disclosed, particularly Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys (SEQ ID No:2). See, for example, US 5,616,311 and WO 96 / 39128.

[0011] The use of lysine amino acid residues in the preparation of multiantigenic peptides has been disclosed in, for example, TamProc.Natl.Acad.,Sci.USA,1988,85,5409-5413, Rao et al.,J.Am.Chem.Soc.,1994,116,6975-6976,US 5,229,490 and WO 2010 / 038220.

[0012] The use of peptide-based scaffolds as drug delivery mediators has been disclosed. See, for example, Brokx et al., J. Control. Release, 2002, 78, 115-123.

[0013] There is clearly a need for new and / or improved medicines that can be used to treat inflammation and conditions characterized by inflammation. Summary of the Invention

[0014] According to a first aspect of the invention, a compound of formula I is provided herein.

[0015] AQB I

[0016] in:

[0017] A and B independently represent Z or A. 1 -Q 1 -B 1 ;

[0018] Q represents a structural fragment of formula II.

[0019]

[0020] in:

[0021] The wavy line represents the attachment point of Q to A and / or B; and

[0022] m represents an integer from 1 to 4;

[0023] A 1 and B 1 Independently represent Z or A 2 -Q 2 -B 2 ;

[0024] A 2 and B 2 Independently represent Z or ZQ 3 -Z;

[0025] Q 1 Q 2 and Q 3 Independently representing the structural fragment of Formula III,

[0026]

[0027] in:

[0028] The wavy lines adjacent to the NH group represent Q. 1 Q 2 and Q 3 With A 1 and / or B 1 A 2 and / or B 2 The attachment point of Z; and the wavy lines adjacent to the O atom represent Q respectively. 1 Q 2 and Q 3 With Q, Q 1 and Q 2 The attachment point; and m is as defined above;

[0029] In each case where it is used, Z represents a peptide component having the following amino acid sequence:

[0030] [W-Lys-X 1 -Ser-UX 2 -Y] n -W-Lys-X 1 -Ser-UX 2 -Y---(SEQ ID No:3)

[0031] in:

[0032] The dashed lines represent the attachment points of Z to the rest of the molecule;

[0033] n represents 0 or an integer from 1 to 4; and

[0034] In each case where they are used:

[0035] W represents a 1 or 2 amino acid sequence, wherein the amino acid is selected from one or more of the group consisting of Lys, Ala, DOPA and 3,4-dihydrocinnamic acid (HCA) residues, provided that, when present, the HCA residue is located at the N-terminus of the peptide sequence Z.

[0036] X 1 Represents Pro, Hyp, or diHyp;

[0037] U stands for Tyr or DOPA;

[0038] X 2 Represents Ser, Pro, Hyp, or diHyp; and

[0039] Y represents a sequence of 1 to 5 (e.g., 1 to 4) amino acids, wherein the amino acids are selected from one or more of the group consisting of: Lys, Ala, Pro, Hyp, diHyp, Thr, DOPA, and Tyr.

[0040] And the regioisomers, stereoisomers, and pharmaceutically or cosmetically acceptable salts of the compound.

[0041] The compounds, regioisomers, stereoisomers, and salts mentioned herein are collectively referred to as "the compounds of this invention".

[0042] The compounds of the present invention that may be mentioned include these compounds, wherein:

[0043] W represents a 1 or 2 amino acid sequence, wherein the amino acid is selected from one or more of the following groups: Lys, Ala and DOPA;

[0044] X 1 It stands for Pro;

[0045] X 2 Represents Ser, Pro, or Hyp;

[0046] Y represents a sequence of 1 to 5 (e.g., 1 to 4) amino acids, wherein the amino acids are selected from one or more of the following groups: Lys, Ala, Pro, Hyp, Thr, DOPA, and Tyr.

[0047] Preferred compounds of the present invention that may be mentioned are those in which m represents 1, 3, or more preferably 4, such that Q, Q 1 Q 2 and Q 3One or more of the representatives Lys, or more appropriately, the "Lys segments" (where appropriate) as defined above as "structural segments of Equations II and III".

[0048] In each case where they are used, Q, Q 1 Q 2 and Q 3 Each can be attached to zero, one, or two Z groups.

[0049] In this respect, preferred compounds of the present invention include these compounds, wherein: one of A or B represents Z, and the others represent A. 1 -Q 1 -B 1 Or more preferably, A and B both represent Z or both represent A. 1 -Q 1 -B 1 ,

[0050] In each case, Q 1 Preferably, it represents the Lys fragment, and Z is as defined above.

[0051] Further preferred compounds of the present invention include those compounds, wherein:

[0052] A 1 and B 1 One represents Z, and the others represent A. 2 -Q 2 -B 2 Or more preferably,

[0053] A 1 and B 1 Both represent Z or both represent A 2 -Q 2 -B 2 ,

[0054] In each case, Q 2 Preferably, it represents the Lys fragment, and Z is as defined above.

[0055] Further preferred compounds of the present invention include those compounds, wherein:

[0056] A 2 and B 2 One represents Z, and the others represent ZQ. 3 -Z; or more preferably,

[0057] A 2 and B 2 All represent Z or all represent ZQ 3 -Z,

[0058] In each case, Q 3 Preferably, it represents the Lys fragment, and Z is as defined above.

[0059] More preferably, the compound of the present invention includes A. 2 and B 2 Those that all represent Z.

[0060] The peptide components of the compounds of the present invention that may be mentioned include those in which n is 0, 1, or 4, or more preferably n is 0.

[0061] Preferred compounds of the present invention include these compounds, wherein:

[0062] X 1 Represents Hyp or more preferably Pro;

[0063] X 2 Represents Hyp;

[0064] W represents HCA, HCA-Ala-, preferably Ala or Lys-Ala or more preferably DOPA or DOPA-Ala-; and / or

[0065] Y represents a 5, preferably 3 or more preferably 4 amino acid sequence, wherein the amino acid is selected from one or more of the group consisting of Lys, Ala, Hyp, Thr, DOPA and Tyr.

[0066] More preferably, the compounds of the present invention further include those wherein Y represents a 4-amino acid sequence selected from the group consisting of: -Pro-Y 1 -Y 2 -Lys-, or more preferably, -Hyp-Y 1 -Y 2 -Lys- and -Thr-Y 1 -Y 2 -Lys, where Y 1 and Y 2 Each is independently selected from the following groups: Pro, or more preferably, Ala, Hyp, Thr, DOPA, and Tyr.

[0067] Further preferred compounds of the present invention include those wherein the amino acid sequence defined by Y is selected from the group consisting of:

[0068] -Pro-Thr-DOPA-Lys-;

[0069] -Pro-Thr-Tyr-Lys-;

[0070] -Thr-Tyr-Pro-Lys-; and

[0071] -Thr-DOPA-Pro-Lys-; and more preferably,

[0072] -Hyp-Thr-Tyr-Lys-;

[0073] -Hyp-Thr-DOPA-Lys-;

[0074] -Hyp-Thr-Ala-Lys-;

[0075] -Thr-Tyr-Hyp-Lys-;

[0076] -Thr-DOPA-Hyp-Lys-; and

[0077] -Thr-Ala-Hyp-Lys-.

[0078] When Y represents a 2-amino acid sequence, the preferred compounds of the present invention include those wherein the amino acid sequence defined by Y is selected from the group consisting of -Hyp-Thr-, -Thr-Tyr-, -Pro-Thr-, and -Thr-DOPA-.

[0079] Other preferred compounds of the invention may be mentioned, wherein the amino acid sequence defined by Y is selected from the group consisting of -Thr-Tyr-Lys-, -Tyr-Pro-Lys-, -DOPA-Pro-Lys-, -Hyp-Thr-Tyr-, -Hyp-Thr-Tyr-Hyp-Lys-, and more preferably the group consisting of -Thr-Tyr-Hyp-Lys-DOPA- and -Hyp-Thr-DOPA-.

[0080] The compounds of the present invention that may be mentioned include these compounds, wherein:

[0081] U stands for Tyr; and / or

[0082] W stands for Ala.

[0083] In this regard, further compounds of the invention may be mentioned, including those compounds wherein Z is selected from the group consisting of:

[0084] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 2);

[0085] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 1);

[0086] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 4);

[0087] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 5);

[0088] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys-DOPA---(SEQ ID No: 6);

[0089] Ala-Lys-Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys--(SEQ ID No: 7); and

[0090] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp -Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No:8).

[0091] The compounds of the present invention that may be mentioned include these compounds, wherein:

[0092] U stands for Tyr; and / or

[0093] W stands for Lys-Ala-.

[0094] In this regard, further compounds of the invention may be mentioned, including those compounds wherein Z is selected from the group consisting of:

[0095] Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr---(SEQ ID No: 9);

[0096] Lys-Ala-Lys-Hyp-Ser-Tyr-Hyp-Hyp-Thr-DOPA---(SEQ ID No:10); and more preferably,

[0097] Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA---(SEQ ID No: 11).

[0098] Further compounds of the invention that may be mentioned include these compounds, wherein: U represents Tyr; and / or

[0099] W stands for HCA, HCA-Ala-, or more preferably DOPA or DOPA-Ala-.

[0100] In this regard, the compounds of the present invention that may be mentioned include those compounds, wherein Z is selected from the group consisting of:

[0101] HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 12);

[0102] HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 13);

[0103] HCA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 14);

[0104] HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 15);

[0105] More preferably, Z is selected from the group consisting of:

[0106] DOPA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 16);

[0107] DOPA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 17);

[0108] DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 18); and DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 19).

[0109] Other compounds of the invention that may be mentioned include those in which U represents DOPA.

[0110] In this regard, the compounds of the present invention that may be mentioned include those compounds, wherein Z is selected from the group consisting of:

[0111] Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 20);

[0112] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 21);

[0113] Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA---(SEQ ID No: 22);

[0114] Lys-Ala-Lys-Hyp-Ser-DOPA-Hyp-Hyp-Thr-DOPA---(SEQ ID No: 23);

[0115] More preferably, Z is selected from the group consisting of:

[0116] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 24); and

[0117] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp -Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ IDNo:25).

[0118] Further compounds of the invention that may be mentioned include these compounds, wherein: U represents DOPA; and / or

[0119] W stands for HCA, HCA-Ala-, or more preferably DOPA or DOPA-Ala-.

[0120] Therefore, the specific compounds mentioned in this invention include those compounds, wherein Z is selected from the group consisting of:

[0121] HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 26);

[0122] HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 27);

[0123] HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 28);

[0124] HCA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 29);

[0125] More preferably, Z is selected from the group consisting of:

[0126] DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 30);

[0127] DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 31);

[0128] DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 32);

[0129] DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No:33); and

[0130] DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 34).

[0131] The compounds of the present invention that may be mentioned include these compounds, wherein:

[0132] Both A and B represent Z;

[0133] One or preferably two Z groups represent:

[0134] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No:4),

[0135] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No:5),

[0136] HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 12),

[0137] HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No:15),

[0138] DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys—(SEQ ID No: 18),

[0139] Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 20),

[0140] Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA---(SEQ ID No: 22),

[0141] HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 26),

[0142] DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No:31),

[0143] DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No:32),

[0144] Or more preferably, one or more preferably two Z groups represent:

[0145] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No:4), or

[0146] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 5);

[0147] Or even more preferably, one or more Z groups represent:

[0148] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No:2), or

[0149] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 1),

[0150] and

[0151] Q represents the Lys fragment.

[0152] Further compounds of the invention that may be mentioned include these compounds, wherein:

[0153] Both A and B represent A. 1 -Q 1 -B 1 ;

[0154] A 1 and B 1 All represent Z;

[0155] One or preferably two Z groups represent:

[0156] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No:4),

[0157] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No:5),

[0158] Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA---(SEQ ID No: 11),

[0159] HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 12),

[0160] HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 13),

[0161] DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 18),

[0162] Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 20),

[0163] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 21),

[0164] Lys-Ala-Lys-Hyp-Ser-Tyr-Hyp-Hyp-Thr-DOPA---(SEQ ID No: 10),

[0165] HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 28),

[0166] DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No:30),

[0167] DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No:32),

[0168] Or more preferably, one or more preferably two Z groups represent:

[0169] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No:2), or

[0170] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No:1); and

[0171] Q 1 Represents the Lys fragment.

[0172] Further compounds of the invention that may be mentioned include these compounds, wherein:

[0173] Both A and B represent A. 1 -Q 1 -B 1 ;

[0174] A 1 and B 1 All represent A 2 -Q 2 -B 2 ;

[0175] A 2 and B 2 All represent Z;

[0176] One or preferably two Z groups represent:

[0177] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No:4),

[0178] Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No:5),

[0179] HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 12),

[0180] DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 18),

[0181] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 21),

[0182] DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No:32),

[0183] Or more preferably, one or more preferably two Z groups represent:

[0184] DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 19),

[0185] Or even more preferably, one or more Z groups represent:

[0186] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No:2), or

[0187] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No:1); and

[0188] Q 1 and Q 2 All of these represent Lys segments.

[0189] Further compounds of the invention that may be mentioned include these compounds, wherein:

[0190] Both A and B represent A. 1 -Q 1 -B 1 ;

[0191] A 1 and B 1 All represent A 2 -Q 2 -B 2 ;

[0192] A 2 and B 2 All represent ZQ 3 -Z;

[0193] One or preferably two Z groups represent:

[0194] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No:2), or

[0195] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No:1); and

[0196] Q 1 Q 2 and Q 3 All of these represent Lys segments.

[0197] According to a further aspect of the present invention, a (separated) peptide compound is provided having the following amino acid sequence:

[0198] [Ala-Lys-X 1 -Ser-UX 2 -Y] p -Ala-Lys-X 1 -Ser-UX 1 -YG(SEQ ID No:35)

[0199] in

[0200] p represents an integer from 1 to 4;

[0201] G may be absent (in which case Y is a C-terminal amino acid), or G may represent DOPA or dopamine (or more appropriately, a "dopamine fragment"); and

[0202] X 1 、U、X 2 Y is as defined above.

[0203] And the regioisomers, stereoisomers, and pharmaceutically or cosmetically acceptable salts of the compound.

[0204] The compounds, regioisomers, stereoisomers, and salts mentioned herein are collectively referred to as "the linear long-chain compounds of the present invention".

[0205] As used herein, the terms dopamine and dopamine fragment refer to the structural fragment of formula IV.

[0206]

[0207] The wavy line represents the attachment point with respect to Y.

[0208] In the linear long-chain compounds of the present invention, the preferred values ​​of p are arranged in ascending order of 2, 3, 1 and 4.

[0209] The preferred values ​​of U, X, and Y mentioned above for the compounds of the present invention are also preferred for the linear long-chain compounds of the present invention.

[0210] The specific linear long-chain compounds of the present invention that may be mentioned are those in which G is absent.

[0211] In this regard, specific linear long-chain peptide compounds include those having the following amino acid sequence: Ala-Lys-Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Ser-Hyp-Thr-Tyr-Lys (SEQ ID No:36);

[0212] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-H yp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys(SEQ ID No:37);

[0213] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys(SEQ ID No:38);

[0214] Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys(SEQ ID No:39);

[0215] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys(SEQ ID No:40);

[0216] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys(SEQ ID No:41); Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys(SEQ ID No:42); and

[0217] Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hy p-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys(SEQ ID No:43).

[0218] To avoid any doubt, the compounds of the present invention as defined above, whether compounds of Formula I or linear long-chain peptide compounds of SEQ ID No:35, are collectively referred to as "compounds of the present invention" in the following text.

[0219] As used herein, Pro represents proline, Ala represents alanine, Ser represents serine, Tyr represents tyrosine, Hyp represents hydroxyproline (including 3-hydroxyproline (3Hyp) and 4-hydroxyproline (4Hyp)), diHyp represents dihydroxyproline (including 3,4-dihydroxyproline (3,4diHyp), 3,5-dihydroxyproline (3,5diHyp), and 4,5-dihydroxyproline (4,5diHyp)), Thr represents threonine, Lys represents lysine, Ala represents alanine, DOPA represents 3,4-dihydroxyphenylalanine, Orn represents ornithine, and Dap represents diaminopropionic acid. The 3,4-dihydrocinnamic acid (HCA) residue is essentially a DOPA residue, but unlike carboxylic acids attached to N-terminal amino acids (Lys or Ala), it lacks a -NH2 group at the 2- or α-carbon position.

[0220] The compounds of the present invention, whether in salt form or other forms, include regioisomers within the amino acids of a peptide (e.g., diHyp, Hyp, and Tyr moieties), and mixtures of such regioisomers. For example, Tyr is defined to include not only tyrosine (4-hydroxyphenylalanine) but also 2- and 3-hydroxyphenylalanine. Hyp is defined to include 4-hydroxyproline (4Hyp), 3-hydroxyproline (3Hyp), and 5-hydroxyproline (5Hyp). More preferably, the Hyp residue is 4-hydroxyproline. Similarly, diHyp is defined to include 3,4-dihydroxyproline (3,4diHyp), 3,5-dihydroxyproline (3,5diHyp), and 4,5-dihydroxyproline (4,5diHyp). More preferably, the diHyp residue is 3,4-dihydroxyproline (3,4diHyp).

[0221] In addition to the standard central carbon atom of the amino acids in the compounds of the present invention (which is generally, but not exclusively, L-configured), certain amino acids in the sequence contain additional chiral carbon atoms. All such stereoisomers and mixtures thereof (including racemic mixtures) are included within the scope of the present invention. In this regard, 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, but it is preferred that the Hyp used in the compounds of the present invention is 4-hydroxy-L-proline. Similarly, the corresponding definitions can be applied to diHyp, where the two hydroxyl groups may also be cis or trans relative to each other. In any case, individual enantiomers of compounds of Formula I that can form part of the compounds of the present invention (as well as the isolated peptide sequences of SEQ ID Nos: 4 to 26) are included within the scope of the present invention.

[0222] The compounds of this invention can be in the form of salts. Salts that may be mentioned include pharmaceutically acceptable 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 this invention with one or more equivalents of a suitable acid or base, optionally in a solvent or in a salt-insoluble medium, followed by removing the solvent or medium using standard techniques (e.g., in a vacuum, by freeze-drying or by filtration). Salts can also be prepared by exchanging the counterion of the compound of this invention in salt form with another counterion, for example using a suitable ion exchange resin.

[0223] Preferred salts include, for example, acetates, hydrochlorides, hydrogen sulfates, maleates, methanesulfonates, toluenesulfonates, alkaline earth metal salts (such as calcium and magnesium salts), or alkali metal salts (such as sodium and potassium salts). Most preferably, the compounds of the present invention may be in the form of acetates.

[0224] The compounds of the present invention can be prepared by conventional techniques, such as standard amino acid coupling techniques, using standard coupling agents and solvents, as described below. The compounds of the present invention can be synthesized from available starting materials using appropriate reagents and reaction conditions. In this regard, those skilled in the art may refer in particular to “Comprehensive Organic Synthesis”, B.M. Trost and I. Fleming, Pergamon Press, 1991. Other applicable references include “Heterocyclic Chemistry”, J.A. Joule, K. Mills, and G.F. Smith, 3rd edition, Chapman & Hall; “Comprehensive Heterocyclic Chemistry II”, A.R. Katritzky, C.W. Rees, and E.F. V. Criven, Pergamon Press, 1996; and “Science of Synthesis”, Volumes 9–17 (Heterenes and Related Ring Systems), Georg Thieme Verlag, 2006.

[0225] The compounds of the present invention can be isolated from their reaction mixtures and, if necessary, purified using conventional techniques known to those skilled in the art. Therefore, the methods for preparing the compounds of the present invention as described herein may include isolating and optionally purifying the compounds of the present invention (as a final step).

[0226] Those skilled in the art will recognize that, in the methods described above and below, it may be necessary to protect the functional groups of the intermediate compound with protecting groups. Protection and deprotection of functional groups can be carried out before or after the reaction.

[0227] Protecting groups can be applied and removed using techniques well known to those skilled in the art and described below. For example, standard deprotection techniques can be used to chemically convert the protected compound / intermediate described herein into an unprotected compound. The type of chemistry involved will determine the need for and type of protecting groups, as well as the order in which the synthesis is completed. The use of protecting groups is fully described in “Protective Groups in Organic Synthesis,” 5th edition, TW Greene & P. ​​GMWutz, Wiley-Interscience (2014), the contents of which are incorporated herein by reference.

[0228] The compounds of this invention are intended for use in human and animal medicine. Therefore, although they may also be used in cosmetics and / or as part of medical devices, they are indicated as pharmaceuticals (and / or in veterinary science).

[0229] The compounds of the present invention themselves (and the isolated peptide sequences) may also possess pharmacological activity, but certain pharmaceutically acceptable (e.g., "protected") derivatives of the compounds of the present invention may exist or can be prepared, which may not possess such activity, but may be administered and subsequently metabolized or chemically transformed to form the compounds of the present invention. Such compounds (which may possess some pharmacological activity, provided that this activity is significantly lower than that of the active compounds obtained by their metabolism / transformation) can therefore be described as "prodrugs" of the compounds of the present invention.

[0230] As used herein, references to prodrugs will include compounds that form experimentally detectable amounts of the compounds of the present invention within a predetermined time after administration. All prodrugs of the compounds of the present invention are included within the scope of the present invention.

[0231] When the compounds of the present invention have pharmacological activity, they are particularly useful in the treatment of inflammation.

[0232] The term “treatment of inflammation” includes the treatment of inflammation (regardless of cause) in any organ of the body, including soft tissues, joints, nerves, vascular systems, internal organs, especially mucosal surfaces, particularly the skin, and also includes all such inflammatory disorders or conditions, and / or disorders or conditions characterized by inflammation (e.g., as symptoms).

[0233] Inflammatory disorders and / or conditions can (and typically) be characterized by the activation of immune defense mechanisms that produce effects that are more harmful than beneficial to the host. Such conditions are usually associated with varying degrees of tissue redness or congestion, swelling, edema, hyperthermia, pain (including aching), fluid exudation, itching (pruritus), cell death and tissue destruction, cell proliferation and / or loss of function.

[0234] Inflammatory conditions that may be mentioned include arteritis, diabetes, metabolic syndrome, rosacea, asthma and allergies, ankylosing spondylitis, chronic obstructive pulmonary disease (COPD), gouty arthritis, inflammatory bowel diseases (such as Crohn's disease and ulcerative colitis), multiple sclerosis, osteoarthritis, pancreatitis, prostatitis, psoriatic arthritis, rheumatoid arthritis, tendinitis, bursitis, Sjögren's syndrome, systemic lupus erythematosus, uveitis, urticaria, vasculitis, mastocytosis, diabetic vascular complications, migraines, atherosclerosis, and related cardiovascular disorders. A disease state characterized by inflammation that may be mentioned is chronic obstructive pulmonary disease (COPD). Other disease states characterized by inflammation that may be mentioned are inflammatory bowel diseases, including Crohn's disease and, especially, ulcerative colitis. Other disease states characterized by inflammation that may be mentioned are gynecological conditions such as cervicitis, vaginitis (e.g., radiation vaginitis), and colpitis. Diseases affecting the gastrointestinal tract, such as gastric ulcers (e.g., gastritis, gastric ulcers, gastric cancer, and other gastric mucosal diseases), as well as gastroesophageal reflux disease (GERD), constipation, and gastritis; inflammation associated with cancer and infections (e.g., viral infections such as the common cold or influenza).

[0235] Inflammatory conditions that may be more specifically mentioned include inflammation of the skin or mucous membranes (including the mucous membranes of the mouth, nose, eyes, vagina, cervix, and / or anorectal region, especially the mucous membranes of the mouth or nose), such as inflammation caused by infections (such as viral and / or bacterial infections), or allergic / atopic conditions (such as rhinitis (e.g., allergic rhinitis), pharyngitis, periodontitis, gingivitis, dry eye, conjunctivitis (e.g., allergic conjunctivitis), dermatitis, urticaria (hives), and food allergies); and other inflammatory conditions such as herpes, drug eruption, polymorphic light eruption, sunburn, early signs of skin cancer (erythematous dermatitis), pathological alopecia (including post-skin grafting), chemical rashes, psoriasis, erythema multiforme, folliculitis, eczema, and otitis externa. A disease state that may be mentioned is polymorphic light eruption.

[0236] More specifically, the compounds can be used to treat certain conditions characterized by and / or associated with inflammation. Such conditions can include wounds (including abrasions (scratches), incisions (including surgical incisions), lacerations, punctures, avulsions, bruises, and scarring) and burns (including inflammation following burns caused by surgical procedures such as skin grafts) and other conditions (such as hemorrhoids). Wounds can be acute or chronic, and / or can be caused by one or more inflammatory disorders as defined herein.

[0237] Wounds on the skin or mucous membranes can be caused by internal or external physical damage to the membrane surface, or by an underlying physiological disorder (i.e., its symptoms).

[0238] Physical (e.g., "open") wounds can be caused by: sharp objects (cuts, incisions, punctures) or blunt objects / mechanical forces (tears, abrasions, avulsions), physical impacts (bruises), heat or chemicals (burns and blisters), ultraviolet radiation (sunburn), or cold (frostbite). Wounds can be superficial (damage to the epidermis and / or dermis only) or full-thickness wounds (damage beneath the epidermis and / or dermis). In severe cases, subcutaneous and / or submucosal tissues, such as muscles, bones, joints, or even internal organs, may be damaged.

[0239] The compounds of this invention can be used to relieve pain (including dull pain) associated with inflammation and / or wounds. In particular, the compounds of this invention can be used to relieve procedural and / or non-procedural pain. Those skilled in the art should understand that the term "procedural pain" (i.e., surgical pain) refers to acute pain associated with medical research and treatment performed for healthcare purposes. The term "non-procedural" refers to general pain associated with inflammation and / or injury (e.g., pain associated with tooth ulcers, burns, and / or scars) and is not the result of a specific medical intervention.

[0240] The compounds of this invention can not only be used to treat inflammation, pain (including dull pain) and / or pruritus (itching) associated with the wound itself and the healing process, but also to prevent fluid leakage from the wound, the risk of infection, and to prevent physiological reactions (such as scarring and hyperpigmentation) caused by inflammation and / or the wound healing process.

[0241] Scarring is a result of inflammation and / or wound healing, and is a general term for the formation of fibrous tissue as a result of such inflammation / healing.

[0242] The compounds of this invention can also be used to inhibit the production of melanin, which may or may not be caused by inflammation and / or wound healing. The compounds of this invention can also be used to inhibit disorders associated with melanin deposition, such as melasma, freckles, melanosis, facial rashes and other pigmentation disorders, skin cancer with melanoma, and pigmentation disorders or skin diseases (like acne) caused by sun exposure.

[0243] Wounds can also occur as a result of (e.g., inflammatory) diseases or disorders. Such wounds can include blisters and / or ulcers of the skin and mucous membranes. They are common conditions that are often long-lasting and difficult to treat. Skin tissue may frequently be damaged, removed, liquefied, infected, and / or necrotic. Ulcers can lead to secondary health consequences (especially if they are infected), are difficult to heal, and are expensive to treat. They can also cause significant psychological stress and financial burden on patients, thus affecting overall well-being and quality of life.

[0244] In alternatives, inflammatory skin conditions or diseases in which the compounds of the present invention are found to be particularly useful include psoriasis, acne, eczema, and dermatitis, especially allergic dermatitis / atopic dermatitis, as well as in the treatment of mucosal inflammations such as, for example, rhinitis (especially allergic rhinitis), hemorrhoids, chronic obstructive pulmonary disease, and ulcerative colitis.

[0245] Psoriasis is a chronic inflammatory skin disease with a tendency to relapse (and some patients never recover throughout their lives). The clinical manifestations of psoriasis primarily include erythema and scaling. It can occur all over the body, but is more commonly observed on the scalp and extremities.

[0246] Acne is a follicular (pilosebaceous unit) chronic inflammatory skin disease that is closely associated with major factors such as excessive sebum production, blockage of the pilosebaceous duct (including closed comedones and open comedones), bacterial infection, and inflammatory response. It tends to occur in adolescence and is characterized by polymorphic skin lesions on the face. Therefore, the term acne includes acne vulgaris and rosacea (i.e., rhinophyma).

[0247] Eczema is an inflammatory skin reaction characterized by intense itching, caused by a variety of internal and external factors. It has three phases: acute, subacute, and chronic. In the acute phase, there is a tendency for exudation, while the chronic phase includes infiltration and hypertrophy. Skin lesions are typically itchy and prone to recurrence.

[0248] Dermatitis is a common skin condition characterized by roughness, redness, itching, eczema, and dryness. If left untreated, small patches, persistent ulcers, and pigmented spots caused by dermatitis can develop into basal cell carcinoma, squamous cell carcinoma, and malignant melanoma. Dermatitis can be caused by a variety of internal and external infectious or non-infectious factors, including substances (contact dermatitis) or allergies (anaphylactic / atopic dermatitis). This also includes seborrheic dermatitis (seborrheic eczema) and all forms of steroid-dependent dermatitis (including photosensitive seborrheic dermatitis, perioral dermatitis, rosacea-like dermatitis, steroid-induced rosacea, steroid-induced rosacea, iatrosacea, rosacea-like steroid dermatitis, topical corticosteroid-induced rosacea-like dermatitis, and more particularly facial corticosteroid-addictive dermatitis (FCAD) or facial corticosteroid-dependent dermatitis (FCDD), characterized by flushing, erythema, telangiectasia, atrophy, papules and / or pustules in the facial area after prolonged use (including uncontrolled use, abuse or misuse) of topical corticosteroids; see, for example, Xiao et al., J. Dermatol., 2015, 42, 697-702 and Lu et al., Clin. Exp. Dermatol., 2009, 35, 618-621.

[0249] Rhinitis is an irritation and inflammation of the mucous membranes inside the nose. Common symptoms of rhinitis include nasal congestion, runny nose, sneezing, and postnasal drip. The most common type of rhinitis is allergic rhinitis caused by allergens such as pollen, dust, mold, or dander from certain animals. Surprisingly, it has been found that even when the compounds of the present invention are applied nasally (i.e., to the nasal mucosa), patients with allergic rhinitis treated with the compounds of the present invention experience relief from eye itching.

[0250] Hemorrhoids are swelling caused by inflammation of the blood vessels inside or around the rectum and anus. Symptoms include bleeding after bowel movements (i.e., injury), prolapsed hemorrhoids, mucus discharge, and itching, pain, redness, and swelling in the anal area. Hemorrhoids are thought to be a result of increased abdominal pressure, such as as a result of constipation or diarrhea.

[0251] Chronic obstructive pulmonary disease (COPD) is a 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 and leads to impaired airflow into and out of the lungs. Symptoms of COPD include shortness of breath, expectorant 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 fumes.

[0252] The compounds of this invention can have a positive effect in alleviating erythema, redness and swelling, edema, blisters and bullous pemphigoid caused by various conditions, including those mentioned herein, and can inhibit the exudation of subcutaneous tissue fluid and suppress itching and pain caused by such inflammatory conditions.

[0253] Other inflammatory conditions that may be mentioned include:

[0254] (a) Mucosal inflammation, such as oral mucositis, aphthous ulcers, otitis media, laryngitis, tracheitis, esophagitis, gastritis, enteritis and enterocolitis (including bacterial dysentery, chronic amoebic dysentery, schistosomiasis, nonspecific ulcerative colitis and regional enteritis), cervicitis and cervical endometritis, endometritis, inflammation caused by inhalation injury, etc., and mucosal inflammation associated with cancers and infections (e.g., viral infections, such as the common cold or influenza) affecting mucosal surfaces (such as those in the mouth, nasopharynx, ear, throat, trachea, gastrointestinal tract, cervix, etc.).

[0255] (b) Orthopedic inflammation associated with, for example, fractures, purulent infections of bones and joints, inflammation caused by rheumatic bone diseases and purulent osteomyelitis (acute, chronic, local, sclerotic, post-traumatic), purulent arthritis; bone tumors (osteosarcoma, osteoid osteoma, chondroma), bone cysts, osteoclastoma, primary osteosarcoma (osteosarcoma, chondrosarcoma, osteofibrosarcoma, Ewing sarcoma, non-Hodgkin's lymphoma, myeloma, chordoma), metastatic bone tumors, bone tumor-like lesions (bone cysts, aneurysmal bone cysts, eosinophilic granulomas, fibrous dysplasia); and rheumatic arthritis.

[0256] (c) Neurological inflammation, such as peripheral polyneuritis, facial neuritis, peripheral neuritis, subcutaneous neuritis, ulnar neuritis, intercostal neuritis, etc.

[0257] (d) Subcutaneous and submucosal soft tissue inflammation, such as myositis, ligamentitis, tendinitis, panniculitis, cystitis, lymphadenitis, bubonadentitis, tonsillitis, synovitis, fasciitis, and soft tissue inflammation caused by injury, contusion, or tear of muscles, ligaments, fascia, tendons, synovium, fat, joint capsules, and lymphatic tissue.

[0258] (e) Vascular inflammation, such as allergic leukocytoclastic vasculitis, allergic cutaneous vasculitis, polyarteritis nodosa, thrombotic vasculitis, granulomatous vasculitis, lymphocytic vasculitis, vasculitis with abnormal blood composition, and rheumatic vasculitis, as well as vascular inflammation associated with vascular carcinoma caused by allergic leukocytoclastic vasculitis, polyarteritis nodosa, thrombotic vasculitis, granulomatous vasculitis, lymphocytic vasculitis, vasculitis with abnormal blood composition, and rheumatic vasculitis.

[0259] (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 pericarditis, myocarditis, endocarditis, pneumonia, hepatitis, spleen inflammation, nephritis, pancreatitis, cystitis, oophoritis, prostatitis, and gastric ulcers.

[0260] (g) Inflammation of the eye and surrounding area, such as conjunctivitis, keratitis (e.g., acute epithelial keratitis, nummular keratitis, interstitial keratitis, discoid keratitis, neurotrophic keratitis, myxomatous keratitis, herpes simplex keratitis, herpes zoster keratitis, bacterial keratitis, fungal keratitis, Acanthamoeba keratitis, onchocercal keratitis, superficial punctate keratitis, ulcerative keratitis, exposure keratitis, photokeratitis, and acute red eye from contact lenses), optic neuritis, etc.

[0261] (h) Inflammation of the gums and mouth, such as periodontitis, gingivitis, tooth ulcers, etc.

[0262] (i) Inflammation associated with rheumatic diseases, such as rheumatic 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, Sjögren's syndrome, spondyloarthritis, systemic lupus erythematosus, tendinitis, etc.

[0263] The compounds of this invention can also be used to treat certain specific diseases of the digestive system, such as gastroesophageal reflux disease (GERD), characterized by a sour taste in the mouth, nausea, heartburn, painful swallowing and / or sore throat, increased salivation (heartburn), nausea, chest pain, and cough. GERD can cause esophageal damage, including reflux esophagitis (i.e., inflammation of the esophageal epithelium, which may cause ulceration at or around the junction of the stomach and esophagus), esophageal stricture (i.e., persistent esophageal stricture caused by reflux-induced inflammation), Barrett's esophagus (i.e., intestinal metaplasia (i.e., a change in the epithelial cells of the distal esophagus from squamous to columnar intestinal epithelium)) and / or esophageal adenocarcinoma (a form of cancer)).

[0264] The compounds of this invention can also be used to treat certain specific diseases of the respiratory system, such as cystic fibrosis, common interstitial pneumonia, allergic pneumonia, asbestosis, emphysema, pulmonary heart disease, and pulmonary embolism. A specific disease condition that may be mentioned is idiopathic pulmonary fibrosis (IPF).

[0265] Interstitial lung disease (IPF) is a diffuse and fatal interstitial lung disease characterized by alveolar epithelial damage, massive proliferation of pulmonary fibroblasts, and excessive deposition of extracellular matrix, ultimately leading to irreversible lung tissue damage. In the later stages of the disease, subjects with IPF experience respiratory failure and death. The compounds of this invention have been found to be useful for treating IPF and / or alleviating symptoms associated with the disease.

[0266] The compounds of this invention are also particularly useful for treating the following lung and / or fibrotic conditions (whether or not otherwise mentioned herein): pulmonary fibrosis, renal fibrosis, liver fibrosis, silicosis, acute bronchitis, chronic bronchitis, tracheobronchitis, bronchial asthma, status asthmaticus, bronchiectasis, upper respiratory tract infections (including the common cold and influenza), allergic airway inflammation, bacterial pneumonia, viral pneumonia, mycoplasma pneumonia, rickettsial pneumonia, radiation pneumonia, and pneumococcal disease (including Staphylococcus aureus). Streptococcal and Gram-negative bacillus pneumonia, pulmonary candidiasis (including aspergillosis, mucormycosis, histoplasmosis, actinomycosis and nocardiosis), pulmonary fungal infection, cryptococcosis, lung abscess, allergic pneumonia, extrinsic allergic alveolitis, pulmonary eosinophilia (including Leoffer's syndrome and eosinophilia), obstructive emphysema, pulmonary edema, pulmonary tuberculosis, respiratory alkalosis / acidosis, acute lung injury, interstitial lung disease, empyema, pulmonary fibroma and cor pulmonale.

[0267] The compounds of this invention are useful for specific mucosal disorders and diseases, including, for example, anorectal diseases such as diarrhea, hemorrhoids, abscesses, fistulas, fissures, anal pruritus, anal sinusitis, warts, and rectal prolapse; inflammatory bowel diseases, including Crohn's disease, particularly ulcerative colitis; gynecological diseases such as cervicitis, vaginitis, pelvic pain and disorders; and dental diseases such as periodontitis.

[0268] By increasing the production of SOD (superoxide dismutase) and reducing lipid oxidation, the compounds of the present invention can further possess antioxidant activity. Therefore, the compounds of the present invention can be considered to have antioxidant properties.

[0269] The compounds of the present invention may also have antipyretic properties, which allow for the treatment of fever and / or relief of its symptoms; for example, by lowering the body temperature of the subject, this results in a reduction of fever. Therefore, the compounds of the present invention and formulations containing them can be considered antipyretics.

[0270] According to another aspect of the invention, a method for treating inflammation, inflammatory disorders, and / or disorders / conditions characterized by inflammation (e.g., as a symptom) is provided, the method comprising administering the compound of the invention or a salt thereof to a patient in need of such treatment.

[0271] To avoid ambiguity, in the context of this invention, the terms “treatment,” “therapeutic,” and “method of treatment” include therapeutic or palliative treatment for patients in need and preventative treatment and / or diagnosis for patients susceptible to inflammation and / or inflammatory disorders.

[0272] The compounds of the present invention may further possess antiviral properties, such as those that, contrary to the treatment of any symptoms (such as pain and / or inflammation) of any viral infection or disease, allow for the treatment of viral infection by its very nature, i.e., by interfering with viral replication within the host. Such antiviral properties may also allow for the prevention of the onset of such infection or disease, protection of host cells from (e.g., further) viral infection, prevention or cessation of the spread of viral infection or disease (within a single host, or from one host to a new host), or allow for the prevention of viral reactivation after it has been latent in the host.

[0273] According to another aspect of the invention, a method for treating a viral infection is provided, the method comprising administering the compound of the invention or a salt thereof to a patient in need of such treatment.

[0274] Viral infections that may be mentioned include those caused by viruses from the following families: Adenoviridae (e.g., adenovirus), Papillomaviridae (e.g., human papillomavirus), Polyomaviridae (e.g., BK virus; JC virus), Herpesvirusidae (e.g., herpes simplex virus type 1; herpes simplex virus type 2; varicella-zoster virus; Epstein-Barr virus). Human cytomegalovirus; human herpesvirus (type 8); poxviridae (e.g., smallpox); hepatoviridae (e.g., hepatitis B virus); parvoviridae (e.g., parvovirus B19); astroviridae (e.g., human astrovirus); calicivirusidae (e.g., norovirus; Norwalk virus); pituriviridae (e.g., Coxsackievirus, hepatitis A virus; poliovirus; rhinovirus); coronavirusidae (e.g., severe acute respiratory syndrome virus); flaviviridae (e.g., hepatitis C virus; yellow fever virus; dengue virus; West Nile virus; tick-borne encephalitis virus); retroviridae (e.g., human immunodeficiency virus; HIV); clovenviridae (e.g., rubella virus); arenaviridae (e.g., Lassa virus); Bunyaviridae (e.g., hantavirus; Crimean-Congo hemorrhagic fever virus); hantavirus Viruses, including the Filoviridae family (e.g., Ebola virus; Marburg virus; Ravn virus), Orthomyxoviridae family (e.g., influenza viruses, including influenza A viruses (e.g., H1N1 and H3N2 viruses), influenza B viruses, or influenza C viruses), Paramyxoviridae family (e.g., measles virus; mumps virus; parainfluenza virus, respiratory syncytial virus), Rhabdoviridae family (e.g., rabies virus), Hepatitis Viridae family (e.g., hepatitis E virus), Reoviridae family (e.g., rotavirus; circovirus; coltivirus; Banna virus), and viruses not assigned to a family (such as hepatitis D virus).

[0275] Viruses that may be mentioned more specifically include herpes simplex virus type 1 and herpes simplex virus type 2, human papillomavirus, influenza virus and parainfluenza virus.

[0276] The compounds of the present invention may further possess antibacterial and / or bacterial inhibitory properties, such as those contrary to the treatment of any symptoms (e.g., pain and / or inflammation) of any bacterial infection or disease, which allow for the treatment of bacterial infections by their very nature, i.e., by interfering with the growth or proliferation of bacteria in the host. Therefore, the compounds of the present invention can be considered bactericides and / or preferably bacterial inhibitors.

[0277] Such antibacterial properties can also allow for the prevention of the onset of such infection or disease, protection of host cells from (e.g., further) bacterial infection, prevention or cessation of the spread of bacterial infection or disease (within a single host or from one host to a new host), or prevention of the reactivation of bacteria after they have been latent in the host.

[0278] According to another aspect of the invention, a method for treating bacterial infections is provided, the method comprising administering the compound of the invention or a salt thereof to a patient requiring such treatment.

[0279] As disclosed herein, the compounds of the present invention may further possess anticancer properties, such as those that, in contrast to the treatment of any symptoms of cancer (such as pain and / or inflammation), allow for the treatment of cancer in its essence, i.e., by interfering with the cancer itself. Such anticancer properties may also include prevention of the onset of the disease, for example, by treating inflammation and thus preventing its onset.

[0280] According to another aspect of the present invention, a method for treating cancer is provided, the method comprising administering the compound of the present invention or a salt thereof to a patient in need of such treatment.

[0281] Specific cancers that may be mentioned include oral cancer, nasopharyngeal cancer, middle ear cancer, conjunctivitis, laryngitis, pharyngitis, laryngitis, tracheitis, esophagitis, gastritis, enterocolitis, cervicitis, endometritis, and skin cancer caused by oral mucositis, rhinitis, otitis media, conjunctival cancer, laryngeal cancer, tracheal cancer, esophageal cancer, gastric cancer, intestinal cancer, cervical cancer, endometrial cancer, and skin cancer. A specific skin cancer that may be mentioned is basal cell carcinoma.

[0282] "Patients" include reptile patients, bird patients, and preferably mammal patients (especially humans).

[0283] According to the present invention, the compounds of the present invention are preferably administered topically or systemically, for example, orally, intravenously or intra-arterially (including via intravascular and other perivascular devices / dosage forms (e.g., stents)), intramuscularly, through the skin, subcutaneously, via mucosa (e.g., sublingual or buccal), rectum, vagina, intradermally, transdermally, via the nose, via the lungs (e.g., trachea or bronchus), preferably topically, or via any other parenteral route, in the form of a pharmaceutical preparation comprising one or more compounds, in one or more pharmaceutically acceptable dosage forms.

[0284] When the condition to be treated is rhinitis or inflammation caused by a viral infection of the airways (such as an upper respiratory tract infection, such as the common cold and influenza), inhalation (e.g., nasal administration) is particularly useful.

[0285] Lung administration is particularly useful when the condition to be treated is COPD or IPF. Topical application can be enhanced by producing a spray containing the active ingredient, for example by using a powder aerosol or by using appropriate nebulization techniques or devices (such as a nebulizer) with the aid of water mist.

[0286] When the condition to be treated is hemorrhoids or ulcerative colitis, rectal administration is particularly useful, using appropriate means of delivery such as injectable foam solutions or suppositories.

[0287] Administration to the lower gastrointestinal tract can also be achieved via parenteral delivery, particularly via oral delivery, using standard delayed or extended release coating techniques known to those skilled in the art. Specifically, different portions of the upper or lower intestine can be targeted. For example, colonic administration can also be achieved using a drug delivery device that initially targets the colon via oral or parenteral administration.

[0288] In an alternative, the compounds of the present invention can be administered via direct systemic parenteral administration. Such administration can be useful in methods of treating inflammatory and / or fibrotic disorders or conditions of one or more internal organs in a patient.

[0289] The internal organs that may be mentioned include the stomach, intestines, pancreas, liver, spleen, bladder, vascular system, ovaries, prostate, with the heart and kidneys being preferred, and the lungs being even more preferred.

[0290] Visceral organ fibrosis includes acute and / or severe internal fibrosis, characterized by excessive accumulation of fibrous connective tissue (as described above) in and around inflamed or damaged tissue. Therefore, the formulations of the present invention can be used to treat or prevent fibrosis (as described above) and the associated morbidity and mortality. Thus, visceral organ fibrosis (e.g., acute and / or severe) that can be treated with the formulations of the present invention includes fibrosis of the liver, kidneys, lungs, cardiovascular system (including the heart and vascular system), pancreas, spleen, central nervous system (neurofibrosis), myelofibrosis, eyes, vagina, cervix, etc.

[0291] Inflammatory conditions of internal organs include any serious condition (i.e., a condition requiring intensive medical treatment) or any condition that can develop into a serious condition, in which some inflammatory component is evident (because it can be characterized by detectable inflammation), and in which the onset is obvious (or anticipated) and / or life-threatening.

[0292] The inflammatory conditions that may be mentioned include one or more acute disorders or conditions of visceral organs (i.e., one or more conditions requiring immediate medical intervention or conditions that may develop into conditions requiring immediate medical intervention), characterized by inflammation (e.g., as a symptom) in one or more visceral organs (including any of the organs mentioned above), such as acute internal damage. By treating such acute inflammatory disorders, the formulations of the present invention can prevent or halt the development of symptoms (acute or chronic) associated with such conditions, and can also halt the progression of morbidity and / or death associated with such conditions.

[0293] Therefore, acute inflammatory conditions that may be mentioned include conditions such as peritonitis, pancreatitis, colitis, proctitis, gastritis, duodenitis, pharyngitis, GERD, periodontitis, and stomatitis. Specific acute inflammatory conditions that may be mentioned include acute injury to one or more internal organs (including any of those mentioned above), such as acute lung injury, inhalation injury (such as burns), acute respiratory distress syndrome (ARDS), severe acute respiratory syndrome (SARS), and multi-organ inflammation, injury, and / or failure.

[0294] These conditions can be caused by internal or external trauma (such as injury or burns), or by infections such as viruses, bacteria, or fungi.

[0295] For example, proctitis (which includes eosinophilic, gonococcal, and / or ulcerative proctitis) can be caused by inflammatory bowel disease, infection, radiation (e.g., for cancer), medications (such as antibiotics), surgery, or allergic conditions (such as food intolerance).

[0296] For example, multi-organ inflammation, injury, and / or failure can be caused by extensive and / or traumatic external injuries (including traumatic and / or extensive external burns). Traumatic external burns will be understood to include second-degree burns, and more specifically, third-degree and fourth-degree burns. Extensive external burns should be understood to include burns covering at least approximately 10% of the patient's body surface area, for example, at least approximately 15%, including burns covering at least approximately 20%. External (and internal) burns can be caused by exposure to heat, chemicals, etc.

[0297] Acute inflammatory and / or fibrotic conditions can also be caused by sepsis or septic shock, which can be caused by viral, bacterial, or fungal infections. Furthermore, acute lung injury, ARDS, and especially SARS, can be caused by viruses such as coronaviruses, including the novel SARS coronavirus type 2 (SARS-CoV-2).

[0298] Therefore, in addition, one or more of the aforementioned (e.g., acute) inflammatory conditions may (and indeed will likely in some cases) lead to some form of internal tissue damage and / or dysfunction of related internal tissues. Related tissues include (e.g., mucosal) tissues, such as the respiratory epithelium. Such tissue damage can also cause one or more of the fibrotic conditions mentioned above. For example, it is known that SARS disease caused by the novel coronavirus SARS-CoV-2 (2019 coronavirus disease or COVID-19) can lead to fibrosis in many cases, caused by one or more of a variety of factors, including inflammation.

[0299] In this respect, the compounds of the present invention and their salts are particularly useful in treating related inflammatory and / or fibrotic conditions, as such conditions are often characterized by one or more comorbidities. By the term "characterized by comorbidities," we assume that the primary condition in question simultaneously causes (and indeed preferably) one or more other medical conditions, including those described above, which may influence each other and / or overlap in some way.

[0300] Therefore, the following is provided:

[0301] A method for treating at least one inflammatory and / or fibrotic disorder or condition of one or more visceral organs in a patient, the method comprising direct systemic parenteral administration of the compound of the present invention or a pharmaceutically acceptable salt thereof to a patient in need of such treatment;

[0302] • A method for treating a patient with two or more inflammatory and / or fibrotic disorders or conditions of one or more internal organs, the method comprising direct systemic parenteral administration of the compound of the present invention or a pharmaceutically acceptable salt thereof to a patient requiring such treatment; and

[0303] • A method for reducing the incidence of morbidity and / or death associated with or potentially associated with one or more inflammatory and / or fibrotic disorders or conditions of one or more internal organs in a patient, the method comprising direct systemic parenteral administration of the compound of the present invention or a pharmaceutically acceptable salt thereof to a patient in need of such treatment.

[0304] When the compounds of the present invention / their salts are administered directly and parenterally, they can be administered intravenously, intra-arterially, intravascularly, perivascularly, intramuscularly, through the skin and / or subcutaneously, for example by direct injection or by any other parenteral route, in pharmaceutically acceptable dosage forms of the compounds of the present invention or their salts.

[0305] Therefore, pharmaceutically acceptable formulations for such administration may comprise the compound of the present invention mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier, the pharmaceutically acceptable adjuvant, diluent, or carrier being selected appropriately, taking into account the intended route of direct parenteral administration and standard pharmaceutical practice. Such pharmaceutically acceptable carriers may be chemically inert to the active compound and may be free of harmful side effects or toxicity under the conditions of use. Such pharmaceutically acceptable carriers may also impart immediate or modulated release of the compound of the present invention.

[0306] Therefore, formulations for injection may be in the form of aqueous formulations such as suspensions and / or more preferably solutions (e.g., (optionally) buffered aqueous formulations (e.g., solutions), such as formulations containing physiological saline (e.g., solutions), phosphate formulations (e.g., solutions), acetate formulations (e.g., solutions), or borate formulations (e.g., solutions), or lyophilized powders (e.g., injections) that can be reconstituted with a medium such as an aqueous medium prior to use).

[0307] Formulations for injection may include other suitable excipients known to those skilled in the art, such as solvents (e.g., water), cosolvents, solubilizers (e.g., cyclodextrin), wetting agents, suspending agents, emulsifiers, thickeners, chelating agents, antioxidants, reducing agents, antimicrobial preservatives, fillers, and / or protectants.

[0308] Preparations for injection are preferably buffered to a physiologically acceptable pH value (e.g., pH between about 4.5 and about 9.5, such as between about 6 and about 9, such as between about 6.5 and about 8.5) using standard techniques, and / or may further contain a tension modifier (such as sodium chloride).

[0309] While the preferred delivery methods described above for the compounds of the present invention include topical application to sites of inflammation (e.g., mucous membranes (including oral and / or nasal mucosa, lungs, anorectal region and / or colon), or more preferably skin), using appropriate (e.g., pharmaceutically acceptable and topically acceptable) media and / or commercially available formulations suitable for application to the skin and / or appropriate mucosal surfaces, they may also include oral, intravenous, skin or subcutaneous, nasal, intramuscular, intraperitoneal or pulmonary delivery.

[0310] Injection is particularly useful for applying the compounds of the invention in suspension form to, for example, the dermis (e.g., intradermal injection), joint cavity, or eye.

[0311] Intradermal injection (e.g., intradermal administration) is particularly useful for applying the compounds of the invention, in solution or suspension form (e.g., dermal fillers), into the dermis. This is particularly useful as a means of administration for the treatment of melanin deposition as described above, or for the use of the compounds of the invention in the treatment of, for example, wrinkles.

[0312] Injection application is particularly useful for filling (e.g., surgical sites in the nasal cavity, anal fistulas, spaces between the gums and tooth roots, or sinuses). It can also be used for shaping support and / or lubrication.

[0313] The compounds of the present invention are typically administered in the form of one or more pharmaceutical formulations, for example, mixed with (e.g., pharmaceutically acceptable) adjuvants, diluents, or carriers, the form of which may be chosen appropriately taking into account the intended route of administration (e.g., topical application to the relevant mucous membranes (including the lungs) or preferably the skin) and standard pharmaceutical or other (e.g., cosmetic) practices. Such pharmaceutically acceptable carriers may be chemically inert to the active compound and may be free of harmful side effects or toxicity under the conditions of use. Such pharmaceutically acceptable carriers may also impart immediate or modulated release to the compounds of the present invention.

[0314] Suitable pharmaceutical formulations may be commercially available or otherwise prepared according to techniques described in the literature, such as Remington, The Science and Practice of Pharmacy, 22nd edition, Pharmaceutical Press (2012) and Martindale, The Complete Drug Reference, 38th edition, Pharmaceutical Press (2014), and the documents mentioned therein, all relevant disclosures of which are hereby incorporated by reference. In other respects, those skilled in the art can prepare suitable formulations comprising the compounds of the present invention without inventive step using conventional techniques.

[0315] The compounds of the present invention may be in the form of aqueous formulations such as emulsions, suspensions and / or solutions (e.g., (optionally) buffered aqueous formulations (e.g., solutions), formulations such as those containing physiological saline (e.g., solutions), formulations containing phosphate (e.g., solutions), formulations containing acetate (e.g., solutions) or formulations containing borate (e.g., solutions)) or lyophilized powders.

[0316] The compounds of the present invention can be further and / or combined with suitable excipients in alternative embodiments to prepare:

[0317] • Gel formulations (for said gel formulations, suitable gel matrix materials include cellulose derivatives, carbomer and alginate, tragacanth gum, gelatin, pectin, carrageenan, gellan gum, starch, xanthan gum, cationic guar gum, agar, non-cellulose polysaccharides, sugars (such as glucose), glycerol, propylene glycol, vinyl polymers, acrylic resins, polyvinyl alcohol, carboxyvinyl polymers, and especially hyaluronic acid);

[0318] • Detergent (for the detergent, suitable matrix materials include cellulose derivatives, glycerol, non-cellulose polysaccharides, polyethylene glycol and propylene glycol of different molecular weights);

[0319] • Paste or ointment (for the paste or ointment, suitable paste base materials include glycerin, petrolatum, paraffin, polyethylene glycol of different molecular weights, etc.);

[0320] • Creams or foams (for said creams or foams, suitable excipients (e.g., foaming agents) include hydroxypropyl methylcellulose, gelatin, polyethylene glycol of varying molecular weights, sodium lauryl sulfate, sodium fatty alcohol polyoxyethylene ether sulfonate, corn gluten flour, and acrylamide);

[0321] • Powdered aerosols (for said powdered aerosols, suitable excipients include mannitol, glycine, dextrin, dextrose, sucrose, lactose, sorbitol, and polysorbate, such as dry powder inhalers); and / or

[0322] • Liquids for oral or inhalation use, such as water (aerosol) sprays (for said liquids, suitable excipients include viscosity modifiers, such as hyaluronic acid, sugars (such as glucose and lactose), emulsifiers, buffers, alcohols, water, preservatives, sweeteners, flavorings, etc.);

[0323] • Injectable solutions or suspensions (which may be aqueous or other forms, and for which suitable excipients include solvents and cosolvents, solubilizers, wetting agents, suspending agents, emulsifiers, thickeners, chelating agents, antioxidants, reducing agents, antimicrobial preservatives, buffers and / or pH adjusters, fillers, protectants and tension modifiers), and specific injectable solutions or suspensions may be mentioned including dermal fillers (i.e., injectable fillers or soft tissue fillers), particularly when the compounds of the present invention are combined with hyaluronic acid.

[0324] Where appropriate, such formulations may also contain humectants such as glycerol, glycerin, polyethylene glycol, trehalose, petrolatum, paraffin oil, silicone oil, hyaluronic acid and its salts (e.g., sodium and potassium salts), octanoic / caprylic triglycerides, etc.; and / or antioxidants such as vitamins and glutathione; and / or pH adjusters such as acids, bases, and pH buffers. Additionally, surfactants / emulsifiers may be included, such as cetyl alcohol, fatty acids (e.g., stearic acid), sodium lauryl sulfate, sorbitan esters (e.g., sorbitan stearate, sorbitan oleate, etc.), monoacylglycerol esters (such as glyceryl monostearate), polyethoxylated alcohols, polyvinyl alcohol, polyol esters, polyoxyethylene alkyl ethers (e.g., polyoxyethylene sorbitan monooleate), polyoxyethylene castor oil derivatives, ethoxylated fatty acid esters, polyoxyglycerol esters, lauryl dimethylamine oxide, choline... Juice salts (e.g., sodium deoxycholate, sodium cholate), lipids (e.g., fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterols, prenol, glycolipids, polyketides), phospholipids, N,N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide, poloxamer, lecithin, sterols (e.g., cholesterol), glycolipids, polysorbates, etc.); preservatives, such as phenoxyethanol, ethylhexylglycerin, etc.; and thickeners, such as acryloyldimethyl taurate / VP copolymer. Specifically, particularly in cream formulations, stearic acid, glyceryl monostearate, hexadecyl alcohol, sorbitan stearate, cetyl alcohol, caprylic / capric glyceride, etc., may be included.

[0325] The compounds of the present invention, and formulations comprising them (e.g., pharmaceuticals) (e.g., aqueous solutions, gels, creams, ointments, lotions, foams, pastes, and / or dry powders as described above), can be further combined with suitable matrix materials to prepare dressings or therapeutic patches for application to biological surfaces, such as skin or mucous membranes. Thus, such formulations can be used to impregnate matrix materials, such as gauze, nonwoven fabric, or silk paper. Alternatively, therapeutic patches can be, for example, adhesive bandages, face masks, eye masks, hand masks, foot masks, etc.

[0326] Vaseline can be used to apply such dressings to wounds, but we have also discovered that PEG-based ointments (e.g., PEG 400) can be combined with matrix materials to prepare dressings without the use of Vaseline.

[0327] The compounds of the present invention can also be used in combination with solid supports such as nasal dressings (e.g., for preventing nosebleeds), dermal scaffolds (e.g., for wound healing) or artificial bone (e.g., in the case of bone grafts / implants).

[0328] The compounds of this invention can be administered by inhalation in the form of suspension, dry powder, or solution. Suitable inhalation devices include pressurized metered-dose inhalers (pMDIs) (which may be manually actuated or breath-actuated and may be used with or without a standard spacer device), dry powder inhalers (DPIs) (which may be single-dose, multi-dose, and power-assisted), and soft mist inhalers (SMIs) or nebulizers (wherein the aerosol drug is delivered in a fine mist at a slower rate than when the spray is delivered using a pMDI).

[0329] In pMDI, the compounds of the present invention can be administered as a pressurized suspension of micronized particles distributed in a propellant (e.g., an HFA along with excipients such as mannitol, lactose, sorbitol, etc.) or as an ethanol solution to deliver one or more doses between about 20 μL and about 100 μL per actuation. Actuation can be performed manually (e.g., by pressing) or by inhalation (breath actuation), which involves a spring-driven flow triggering system.

[0330] In a DPI (Digital Inhalation Product), the compounds of the present invention can be administered as micronized drug particles (between about 1 μm and about 5 μm in size) within a capsule (alone or blended with a larger particle size inactive excipient (e.g., mannitol), the capsules of which can be pre-loaded or manually loaded into the device. Inhalation from a DPI can depolymerize the drug particles and disperse them within the airway.

[0331] In SMI, the compound of the present invention can be stored as a solution in a cartridge loaded into the device. A spring releases the dose into a micro-pump, such that when a button is pressed, the dose is released, thereby releasing a jet of the drug solution.

[0332] The compounds of the present invention can also be administered in the form of a fine mist of atomized solution using various nebulizers. Nebulizers may include respiratory-enhanced jet nebulizers (in which, with the assistance of a compressor, an airflow moves through a jet to atomize the drug solution); respiratory-actuated jet nebulizers (in which, after inhalation by a patient, an airflow moves through a tube with the assistance of a compressor to atomize the drug solution); ultrasonic nebulizers (in which the vibration of a piezoelectric crystal causes atomization by heating); and vibrating mesh nebulizers (in which a piezoelectric crystal causes a mesh plate to vibrate, causing atomization to deliver very fine droplets while the temperature of the solution does not change significantly during atomization).

[0333] According to another aspect of the invention, a method for preparing a pharmaceutical composition / formulation as defined herein is provided, the method comprising associating a compound of the invention as defined above with one or more pharmaceutically acceptable excipients as defined above.

[0334] The compounds of the present invention can also be combined with one or more growth factors in treatment, said growth factors being selected from platelet-derived growth factors (including platelet-derived growth factor, PDGF), osteosarcoma-derived growth factor (ODGF), epidermal growth factor (EGF), transforming growth factor (TGFα and TGFβ), fibroblast growth factor (αFGF, βFGF), insulin-like growth factor (IGF-I, IGF-II), nerve growth factor (NGF), interleukin-type growth factor (IL-1, IL-3), erythropoietin (EPO), and colony-stimulating factor (CSF).

[0335] According to another aspect of the invention, a (e.g., pharmaceutical) composition is provided comprising the compound of the invention and one or more pharmaceutically acceptable excipients, such as adjuvants, diluents, or carriers. Preferred formulations are suitable for topical application to, for example, mucous membranes (including oral and / or nasal mucosa, lungs, anorectal region, and / or colon) or more preferably the skin, and therefore comprise topically acceptable adjuvants, diluents, or carriers.

[0336] Therefore, pharmaceutical compositions comprising the compounds of the present invention are further provided that are suitable for, adapted to and / or packaged for, and presented for topical application (e.g., to mucous membranes, including oral and / or nasal mucous membranes, lungs, anorectal regions and / or colon, or preferably to the skin), and the use of such formulations in treating disorders comprising inflammation, inflammatory disorders and / or conditions characterized by inflammation (e.g., as symptoms) by means of direct topical application of said formulations (e.g., to mucous membranes, including oral and / or nasal mucous membranes, lungs, anorectal regions and / or colon, or preferably to the skin).

[0337] Regarding this aspect of the invention, for the avoidance of doubt, topical formulations containing the compounds of the invention can be used for any and all conditions described herein, including (as mentioned, defined, or described above) in the treatment of any and all inflammatory disorders and / or in the treatment of any and all disorders characterized by inflammation. Similarly, any and all topical formulations containing the compounds of the invention mentioned herein include any and all of those mentioned, defined, or described herein. Any and all of the relevant disclosure herein, in connection with this aspect of the invention, is hereby incorporated by reference.

[0338] Topical formulations containing the compounds of the present invention (e.g., liquid-based or solution-based formulations, e.g., aqueous) can be particularly used for wound healing and can relieve pain (including dull pain) and, in particular, pruritus / itching associated with the wound itself and the wound healing process. Such topical formulations containing the compounds of the present invention can be particularly used to prevent and / or inhibit fluid exudation from the wound, especially during the acute inflammatory phase, such as during the first 48 hours after a burn or wound. This prevents the risk of infection and other physiological reactions. Such topical formulations containing the compounds of the present invention can also be particularly used to prevent and / or inhibit scarring and hyperpigmentation (see above), whether or not related to the wound or other aspects.

[0339] The compounds of this invention can be applied continuously or intermittently. The method of application can also be determined by the timing and frequency of application, but in the case of therapeutic treatment of inflammation, it also depends on the severity of the condition.

[0340] Depending on the impairment, the patient to be treated, and the route of administration, the compounds of the present invention can be administered to patients in need at different therapeutically effective doses.

[0341] Similarly, the amount of the compounds of the present invention in the preparation will depend on the severity of the condition and the patient to be treated, but can be determined by a person skilled in the art.

[0342] In any case, a medical practitioner or other technician will be able to routinely determine the actual dosage, which will be most suitable for the individual patient, depending on the severity of the condition and the route of administration. The dosages mentioned herein are examples of average cases; of course, it is natural that there may be individual cases with higher or lower dosage ranges, and such cases are within the scope of this invention.

[0343] The dosage can be administered between once and four times a day (e.g., three times a day).

[0344] In all cases, the appropriate concentration of the compounds of the present invention in aqueous products, calculated as free (non-salt) compounds, can be from about 0.01 (e.g., about 0.1) to about 15.0 mg / mL.

[0345] In all cases, the appropriate topical dose of the compounds of the present invention, calculated as free (non-salt) compounds, is from about 0.05 μg to about 50 μg / cm³. 2 Treatment area, such as about 0.1 μg (e.g., about 0.5 μg) to about 20 μg / cm². 2 Treatment area, ranging from approximately 1 μg to approximately 10 μg / cm² 2 Treatment area, such as approximately 5 μg / cm² 2 Within the range of the processing area.

[0346] Appropriate doses of the compounds of the invention for nasal administration (e.g., by inhalation) are in the range of about 0.01 μg to about 2000 mg, for example, between about 0.1 μg and about 500 mg, or between 1 μg and about 100 mg. Specific doses for nasal administration that may be mentioned include doses between about 10 μg and about 1 mg, particularly about 0.1 mg (i.e., about 100 μg). Daily nasal administration of about 0.1 mg of the compounds of the invention has been found to be particularly effective in the treatment of conditions associated with inflammation of the nasal passages and mucous membranes (such as rhinitis (e.g., allergic rhinitis)) and / or conditions associated with sinus surgery.

[0347] Appropriate doses of the compounds of the invention for pulmonary administration (e.g., by inhalation) are in the range of about 0.01 μg to about 2000 mg, for example, between about 0.1 μg and about 500 mg, or between 1 μg and about 100 mg. Specific doses for pulmonary administration that may be mentioned include doses between about 10 μg and about 10 mg, particularly doses between about 0.6 mg (i.e., 60 μg) and 6 mg (e.g., for the treatment of COPD or IPF).

[0348] We prefer that the pH value of the formulation containing the compound of the present invention is in the range of about 1.0 to about 9.0 (e.g., about 3.0 to about 8.0).

[0349] In any case, within the context of this invention, the dosage administered to mammals, particularly humans, should be sufficient to produce a therapeutic response in mammals within a reasonable timeframe (as described above). Those skilled in the art will recognize that the exact dosage and composition, as well as the selection of the most appropriate delivery method, will also be particularly influenced by: the pharmacological properties of the formulation, the nature and severity of the condition being treated, the recipient's physical condition and mental acuity, the age, condition, weight, sex, and response of the patient to be treated, the stage / severity of the disease, and genetic differences between patients.

[0350] The compounds of this invention can be used in human and animal medicine. In this respect, and as described above, the compounds of this invention that possess appropriate levels of relevant pharmacological (or biological) activity can be used in human and / or animal medicine.

[0351] Certain compounds of the present invention, particularly those of Formula I, and / or linear long-chain compounds of the present invention (preferably where the long-chain compound W represents HCA, HCA-Ala, or more preferably DOPA or DOPA-Ala, and / or U represents DOPA) may have, in addition to the aforementioned biological activities, adhesive properties.

[0352] These adhesion properties stem from the fact that the relevant W and / or U groups can cross-link to form a three-dimensional network.

[0353] The compounds of this invention can adhere to a variety of substrates, including inorganic substrates (such as glass, metal, etc.) and organic substrates (such as biological tissues).

[0354] In this regard, such compounds of the present invention can also be used as wound surface repair products, wound surface protection products, medical bioadhesive products, medical coating products, industrial coating products (e.g., in corrosion protection of ships, electronic equipment, pipelines, etc.), biochemical reagents, medical products, sterilization products, culture dishes for cell culture, etc.

[0355] The compounds of this invention can form a film on the surfaces of various skin and mucous membrane wounds (such as burns, scalds, ulcers, frostbite, and bedsores) to aid in recovery. These compounds can also be used in surgery, for example, for the closure of surgical incisions, adhesion of fractured bone, adhesion of mucous membranes, and as coatings for human implants such as artificial bone, cartilage scaffolds, periosteum, artificial joints, dental implants, occlusive stents, spinal fusion devices, spinal spacers, and organ patches.

[0356] According to another aspect of the invention, compounds of formula I and / or linear long-chain compounds of the invention (preferably wherein the linear long-chain compound W represents HCA, HCA-Ala, or more preferably, DOPA or DOPA-Ala, and U represents DOPA) are provided as adhesives or film-forming materials.

[0357] As discussed above, naturally occurring MAPs are known for their adhesive properties. However, it should be remembered that such adhesive properties may stem from the fact that they are high-molecular-weight linear peptides that can exist in multiple conformations, enabling intermolecular and intramolecular reactions / crosslinking of DOPA residues in the molecule and thus achieving adhesion. In contrast, the compounds of the present invention as defined above are not linear peptides or proteins, but rather, for example, multi-branched lower-molecular-weight residues, and surprisingly, properties similar to those of naturally occurring MAPs (both adhesive and biological properties) have been observed.

[0358] Such crosslinking can be achieved by a variety of chemical means (e.g., iodine vapor, glutaraldehyde, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (EDC / NHS), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine hydrochloride (DMTMM) or other water-soluble condensing agents) or enzymatic means (e.g., tyrosinase or as described below).

[0359] Regardless of the level of pharmacological activity that the compounds of the present invention may have, they can in any case (and / or may further) be combined with an active pharmaceutical ingredient in combination therapy (as described below), or function as a pharmaceutically acceptable excipient (adjuvant, diluent or carrier) or part thereof, as part of a medical device and / or as part of a drug-medical device combination.

[0360] Certain compounds of the present invention can therefore be described as novel multifunctional excipients that can be used in a variety of applications in the pharmaceutical field. In this respect, such compounds of the present invention include those that can be used as adhesives and / or as film-forming agents (as described above), and furthermore, such compounds of the present invention and / or different compounds of the present invention can alternatively and / or additionally be used as delayed-release polymers, as adhesives, as suspending agents, as gelling agents, as coating agents, as diluents, or as carriers of active ingredients (pharmaceuticals) of various solubilities.

[0361] The compounds of the present invention, particularly suitable for use as pharmaceutical excipients, are suitable for large-scale production and may not pose significant toxicity risks, and therefore can be described and listed as “generally recognized as safe” (GRAS) by the U.S. Food and Drug Administration (FDA).

[0362] Such compounds of the present invention can also be used as excipients in veterinary science and cosmetics.

[0363] According to another aspect of the invention, a pharmaceutical formulation is provided comprising an active pharmaceutical ingredient mixed with a pharmaceutically acceptable excipient system (such as a pharmaceutically acceptable adjuvant, diluent, or carrier system), the excipient system comprising one or more compounds of the invention.

[0364] Furthermore, the compounds of the present invention can be combined with active pharmaceutical ingredients and thus can be used as part of a pharmaceutical-medical device combination comprising one or more active pharmaceutical ingredients and one or more compounds of the present invention, wherein the one or more compounds of the present invention constitute a medical device component of the combination.

[0365] When used as a medical device or a component of a pharmaceutical-medical device combination, or as part thereof, those skilled in the art will understand that the relevant compounds of the present invention will be used in human or animal medicine, optionally in combination with an active pharmaceutical ingredient, in a manner that affects the structure and / or one or more functions of the human or animal body, and will achieve their primary intended purpose without the need for chemical action in or on said human or animal body (optionally in a manner that does not depend on the metabolism of the compounds of the present invention to achieve any of their primary intended purpose).

[0366] In this respect, the compounds of the present invention can be combined with a variety of known pharmaceutical active ingredients, and can be combined in this way regardless of whether the compounds of the present invention are used:

[0367] • As a standalone active pharmaceutical ingredient in combination therapy;

[0368] • As a medical device or as part thereof;

[0369] • As a drug-medical device combination or as a medical device component of a drug-medical device combination; or

[0370] • As a pharmaceutically acceptable excipient.

[0371] Such patients may also be (and / or may already be) receiving therapy based on the administration of one or more of these other known pharmaceutical active ingredients, which we mean by receiving a prescription dose 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.

[0372] Pharmaceutical active agents that can be co-administered with the compounds of the present invention include any agent or drug capable of producing a physiological effect (whether therapeutic or preventive against a particular disease state or symptom) in a living subject (including, in particular, mammals and especially, human subjects (patients)).

[0373] Furthermore, the compounds of the present invention (such as those that can be crosslinked as described above) can be used as pharmaceutical excipients and can be mixed with such pharmaceutical active ingredients before or after the crosslinking and / or at least partial crosslinking as described above to form a stable pharmaceutical composition, wherein the compounds of the present invention act as excipients, such as carriers. When used in this manner, it can be found that the compounds of the present invention can actively influence the physical, chemical, and / or biological properties of such active ingredients, including their physical and / or chemical stability and / or their metabolism after administration.

[0374] Pharmaceutical active agents that can be used with the compounds of the present invention may be selected, for example, from anti-inflammatory agents, pro-inflammatory agents, antibiotics, antibacterial agents and / or antibacterial agents, antiviral agents (e.g., protease inhibitors), anesthetics and wound healing agents (e.g., growth factors).

[0375] Bioactive agents may be selected, for example, from anti-inflammatory agents, pro-inflammatory agents, antibiotics, antibacterial agents and / or antibacterial agents, antiviral agents (e.g., protease inhibitors), anesthetics and wound healing agents (e.g., growth factors).

[0376] Non-limiting examples of anti-inflammatory drugs that may be used include anti-inflammatory agents for the treatment of rheumatic diseases and / or arthritis (such as cataflam, betamethasone, naproxen, cyclosporine, chondroitin, celecoxib, etodoxacin, meclofenamic acid, disalicylate, methylprednisolone, and piroxicam); anti-inflammatory agents for the treatment of osteoarthritis (such as sulindac, meloxicam, fenolofen, etodoxacin, and nabumetone); and anti-inflammatory agents for the treatment of inflammation and its symptoms (…). For example, anti-inflammatory agents for fever, pain, itching, and / or swelling (such as mefenamic acid, indomethacin, aspirin, ketorolac, flumethrin, clotiprednisolone, hydrocortisone, flumethrin, bromofenac, prednisolone acetate, indomethacin, and ibuprofen); anti-inflammatory agents used to treat allergies and their symptoms (such as benzimidil, diphenhydramine, naphazoline, antazoline, prednisolone, lodusamide, pyrimisulfan, oxymetazoline, ketotifen, naphazoline, emestine fumarate). fumarate, olopatadine, azelastine, tranilastine, levocabastine, cortisone, ephedrine, cetirizine, levocetirizine, pseudoephedrine, fexofenadine, terfenadine, loratadine, and alexis; anti-inflammatory agents (such as budesonide, cizzonide, nedolome, dexamethasone, ambroxol, and proxetine) used to treat respiratory diseases (including asthma and / or COPD); and drugs used to treat skin diseases. Anti-inflammatory agents for skin diseases (such as mometasone, triamcinolone, desonide, sulfacetamide, tacrolimus, allantoin, and triamcinolone); anti-inflammatory agents for the treatment of mastocytosis (such as cromoglycine); anti-inflammatory agents for the treatment of gout (such as diclofenac and febuxostat); anti-inflammatory agents for the treatment of conjunctivitis (such as hydroxybenzazole, pranoprofen, and zinc sulfate); anti-inflammatory agents for the treatment of eye diseases (e.g., dextran 70, thyroxine / iothlonine, and ocular extractive), any of the above-mentioned known or commercially available pharmaceutically acceptable salts, and any combination of the above-mentioned compounds and / or salts.

[0377] Anti-inflammatory agents that may be mentioned include endogenous (and / or exogenous) lipid-based pro-regression, anti-inflammatory molecules or mediators, such as lipoxygenin, regressors, and protectants. Pro-inflammatory agents that may be mentioned include prostaglandins (e.g., latanoprost, prostacyclin E1, and prostacyclin E2) and leukotrienes (e.g., leukotriene B4).

[0378] Non-limiting examples of antibacterial drugs that may be used include chloramphenicol, ofloxacin, levofloxacin, tobramycin, norfloxacin, ciprofloxacin, lomefloxacin, lincomycin, fluconazole, enoxacin, furazolidone, nitrofurazone, rifampin, fenofibrate, gentamicin, cetirizine, neomycin, roxithromycin, silver sulfadiazine, clarithromycin, clindamycin, metronidazole, azithromycin, sulfamidone, sulfamethoxazole, acetaminophen, chloramphenicol, pseudoephedrine, mupirocin, amoxicillin, amoxicillin / clavulanic acid, trimethoprim / sulfamethoxazole, cephalexin, moxifloxacin, any known or commercially available pharmaceutically acceptable salt of the foregoing drugs, and any combination of the foregoing compounds and / or salts.

[0379] Non-restricted examples of antiviral drugs that may be used include tobramycin, ribavirin, acyclovir, morpholine guanidine, foscarnet, ganciclovir, idoxuridine, trifluuridine, brivudine, vidarabine, entecavir, telbivudine, foscarnet, zidovudine, doxorinosine, zalcitabine, stavudine, lamivudine, abacavir, emtricitabine, nevirapine, deraviridine, efavirenz, etravirine, rilpivirine, saquinavir, ritonavir, indinavir, nelfinavir, amphenavir, lopinavir, ritonavir, atazanavir, fosamprenavir, tepranavir, darunavir, terabhivir, and boceprevir. The following are listed: cimetvir, asunaprevir, raltegravir, elvitegravir, dulutegravir, rsv-igiv, palivizumab, docosanol, enfuvirtide, maraviro, vzig, varizig, acyclovir, ganciclovir, famciclovir, valacyclovir, penciclovir, valganciclovir, cidofovir, tenofovir disoproxil fumarate, adefovir dipivoxil, fomivirsin, prodafiloyl, imiquimod, sinecatechin, interferon-α2b (recombinant, human), any known or commercially available pharmaceutically acceptable salts of the aforementioned drugs, and any combination of the aforementioned compounds and / or salts.

[0380] Non-limiting examples of anesthetic agents that may be used include articaine, dextropropoxyphene, sevoflurane, cophenylcaine, lidocaine, prilocaine, promocaine, benzocaine, dibucaine, diclonine, tetracaine, bupivacaine, and any known or commercially available pharmaceutically acceptable salts of the foregoing drugs, as well as any combination of the foregoing compounds and / or salts.

[0381] Non-limiting examples of wound healing agents that may be used include basic fibroblast growth factor (recombinant, human; recombinant, bovine), epidermal growth factor (recombinant, human; yeast), rhEFG(I), acidic fibroblast growth factor (recombinant, human), granulocyte-macrophage stimulating factor (recombinant, human), silver sulfadiazine, zinc sulfadiazine, fusidic acid, bacitracin, chlorhexidine, silver nitrate, triethanolamine, ethacridine, retinoids, calf blood deproteinized extract, carraghenate, aminoiodide peptide, and any known or commercially available pharmaceutically acceptable salts of the foregoing drugs, as well as any combination of the foregoing compounds and / or salts.

[0382] Such active pharmaceutical ingredients include those that can be applied topically to the skin or mucous membranes, for example, together with the compounds of the present invention. In this regard, preferred active ingredients from the above list include cyclosporine, chondroitin, clotipreno, flumethonone, bromofenac, prednisolone acetate, indomethacin, oxymetazoline, ketotifen, naphazoline, emetine fumarate, olopatadine, azelastine, tranilastine, levocetirizine, cortisone, ephedrine, cetirizine, pseudoephedrine, levocetirizine, fexofenadine, terfenadine, loratadine, alexis, dexamethasone, ambroxol, sulfacetamide, and sulfacetamide. Tacrolimus, allantoin, triamcinolone, cromoglycine, nedocromil, diclofenac, hydroxybenzylazole, pranoprofen, zinc sulfate, dextran 70, thyroxine / iodoxelonine, ophthalmic peptide, chloramphenicol, ofloxacin, levofloxacin, tobramycin, norfloxacin, ciprofloxacin, lomefloxacin, lincomycin, fluconazole, enoxacin, furazolidone, nitrofurazone, rifampin, phenoxybenzoic acid, gentamicin, cetirizine, neomycin, roxithromycin, silver sulfadiazine, clarithromycin Sulfamethoxazole, chloramphenicol, tobramycin, ribavirin, acyclovir, morpholine guanidine, phosphonoformic acid, ganciclovir, interferon-α2b (recombinant, human), articaine, dextropropoxyphene, sevoflurane, cophenylcaine, lidocaine, prilocaine, promocaine, benzocaine, dibucaine, diclonine, tetracaine, bupivacaine, basic fibroblast growth factor (recombinant, human; recombinant, bovine), epidermal growth factor (recombinant, Human; yeast), rhEFG(I), acidic fibroblast growth factor (recombinant, human), granulocyte-macrophage stimulating factor (recombinant, human), silver sulfadiazine, zinc sulfadiazine, fusidic acid, bacitracin, chlorhexidine, silver nitrate, triethanolamine, ethacridine, vitamin A-like substances, calf blood deproteinized extract, carrageenan, aminoiodide peptide, and any known or commercially available pharmaceutically acceptable salts of the foregoing drugs, and any combination of the foregoing compounds and / or salts.

[0383] Other pharmaceutically active ingredients that can be co-administered with the compounds of the present invention include those that can be administered to treat one or more of the gastrointestinal disorders mentioned above.

[0384] Non-restrictive examples of gastrointestinal drugs include oxalazine, sulfasalazine, domperidone, erythromycin, berberine, dexamethasone, cefuroxime axetil, levofloxacin, mesalazine, belladonna, sulfobenzidine, azathioprine, sulfasalazine, live Bacillus species (such as Clostridium butyricum, Bacillus licheniformis, Bacillus cereus), probiotics (such as Bifidobacterium), tegafur, nifuratel, amoxicillin, ampicillin, nystatin, allicin, cefadroxil, dacronin, carmoflurane, fluorouracil, mosapride, thrombin, pantoprazole, cimetidine, cisapride, ethylenediamine diacetamide, nimustine, and famotidine. Barium sulfate, aminocaproic acid, roxatidine acetate, vincristine, azasetron, lentinan, bismuth salts combined with, for example, magnesium salts (e.g., aluminates, potassium citrate), magnesium trisilicate, bicarbonate, vitamin U, aluminum hydroxide, belladonna extract, famotidine and calcium carbonate, magnesium hydroxide, aluminum magnesium carbonate, proton pump inhibitors (e.g., omeprazole, lansoprazole, rabeprazole, pantoprazole, dexlansoprazole, or esomeprazole), glycine, trypsin, allantoin, aluminum hydroxide, sodium L-glutamine guanylate, rebampette, rotundine, quxipite, lafutidine, thymoprotein, Hericium erinaceus The following are listed as pharmaceutically acceptable salts: erinaceus, esolamide maleate, nizatidine, L-glutamine and sodium azulene sulfonate (guanidinium sodium), ranitidine, bismuth citrate, lactobacillus, bisacordine, dimethicone, Clostridium butyricum, loperamide hydrochloride, dibazol, secnidazole, zinc acephate, montmorillonite, tegafur / gemeracil / oteracil, famotidine, oteracil, desoxyfluorouridine, capecitabine, and any of the aforementioned drugs that are known or commercially available.

[0385] Pharmaceutical active ingredients that may be mentioned for use in combination with the compounds of the present invention include active ingredients (other anti-inflammatory agents) that can be used to treat inflammation and / or inflammatory disorders.

[0386] Anti-inflammatory agents that can be used in combination with the compounds of the present invention in the treatment of inflammation include therapeutic agents that can be used to treat inflammation and / or diseases characterized by inflammation as one of their symptoms (including those described above). Depending on the condition to be treated, such anti-inflammatory agents may include NSAIDs (e.g., aspirin), aminosalicylates (e.g., 5-aminosalicylic acid (mesalazine)), leukotriene receptor antagonists (e.g., montelukast, prolucast, and zafirlucast), corticosteroids, analgesics, and certain enzymes (such as trypsin), as described below. The compounds of the present invention may also be combined with leukotrienes (e.g., cysteyl leukotrienes and leukotriene B4).

[0387] Other preferred agents that can be combined with the compounds of the present invention include LTB4 (for treating wounds and burns), NSAIDs (e.g., aspirin) or montelukast (generally used to treat inflammation) and trypsin (for treating mucosal inflammation associated with, for example, viral infections).

[0388] The compounds of the present invention can also be combined with other therapeutic agents, which are known to produce inflammation as a side effect when applied.

[0389] The conjugates of the present invention can also be combined with stem cells (e.g., totipotent stem cells, pluripotent stem cells (such as embryonic or induced pluripotent stem cells), pluripotent stem cells (such as mesenchymal stem cells), oligopotent stem cells (such as hematopoietic stem cells), or unipotent stem cells (such as muscle stem cells)).

[0390] Other known pharmaceutically active ingredients can also be used in combination with the compounds of the present invention in various ways.

[0391] For example, the conjugates of the present invention can be “combined” with (or with other) pharmaceutical active ingredients (or “therapeutic agents”) for use together in the same (e.g., pharmaceutical) formulation, or individually (simultaneously or sequentially) in different (e.g., pharmaceutical) formulations.

[0392] Therefore, such combination products provide for the combined administration of the compounds of the present invention with (or other) therapeutic agents, and can thus be presented as single formulations wherein at least one of these formulations contains the compounds of the present invention and at least one contains (or other) therapeutic agents, or can be presented (i.e., formulated) as combination formulations (i.e., presented as a single formulation comprising the compounds of the present invention and (or other) therapeutic agents).

[0393] Therefore, the following further information is provided:

[0394] (1) A formulation (e.g., a pharmaceutical preparation) comprising the compound of the present invention; another pharmaceutically active ingredient; and optionally, a pharmaceutically acceptable inactive excipient (e.g., an adjuvant, diluent, or carrier), said formulation hereinafter referred to as a “combination formulation”; and

[0395] (2) A kit comprising the following components:

[0396] (A) The compounds of the present invention, optionally in the form of a formulation (e.g., a pharmaceutical preparation) mixed with a pharmaceutically acceptable inactive excipient (e.g., an adjuvant, diluent, or carrier); and

[0397] (B) Another active pharmaceutical ingredient, optionally in the form of a formulation (e.g., a drug) mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier.

[0398] The components (A) and (B) are each provided in a form suitable for combined application with each other.

[0399] In another aspect of the invention, a method for preparing a combination formulation (1) as defined above is provided, the method comprising associating the compound of the invention, other pharmaceutically active ingredients and at least one (e.g., pharmaceutically acceptable) excipient.

[0400] In another aspect of the invention, a method for preparing a kit (2) as defined above is provided, the method comprising associating components (A) and (B). As used herein, references to association are intended to make the two components suitable for and to be administered in combination with each other.

[0401] Therefore, regarding the method for preparing a kit as defined above, which involves "associating" two components with each other, we include the following: the two components of the kit can:

[0402] (i) provided separately (i.e., independently of each other), the individual formulations being subsequently put together for use in combination therapy; or

[0403] (ii) Individual components packaged together and presented as a “combination pack” for use in combination therapy.

[0404] Therefore, a further kit is provided, comprising:

[0405] (I) One of components (A) and (B) as defined herein; and

[0406] (II) Instructions for using the component in combination with another of the two components.

[0407] Regarding the aforementioned kit, although the compounds of the present invention may be provided in the form of a formulation (e.g., a pharmaceutical preparation) mixed with one or more other pharmaceutically acceptable excipients (e.g., adjuvants, diluents, or carriers), they are not provided together with such other pharmaceutically acceptable excipients when the purpose of providing the compounds of the present invention is primarily to function as a medical device or excipient. In any case, it is preferable that the (other) pharmaceutically active ingredients of the kit are provided in the form of a pharmaceutical preparation mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier.

[0408] To provide repeatable dosage, the kits described herein may contain more than one appropriate amount / dosage of the compound of the present invention (e.g., formulations containing it), and / or more than one appropriate amount / dosage of other pharmaceutically active ingredients (e.g., formulations containing it). If there is more than one formulation containing an amount / dosage of any of the aforementioned substances, or more than one amount / dosage of any of the aforementioned substances, such formulations may be the same or different in terms of the amount of any compound, one or more chemical compositions, and / or one or more physical forms.

[0409] Regarding the kits described herein, we use "in combination with" to mean that the corresponding components are administered sequentially, separately, and / or simultaneously during the treatment of the relevant condition.

[0410] Therefore, regarding the combination product according to the invention, the term "in combination with" includes the administration of two components of the combination product (the compound of the invention and other pharmaceutically active ingredients) together or in sufficiently close proximity in time (optionally repeatedly) to enable the patient to receive a beneficial effect during the treatment of the relevant condition, said beneficial effect being greater than if a formulation containing the compound of the invention or (e.g., containing) another agent were administered alone (optionally repeatedly) in the absence of the other component during the same treatment. The determination of whether the combination provides a greater beneficial effect with respect to a specific condition and during its treatment will depend on the condition to be treated or prevented, but can be routinely performed by those skilled in the art.

[0411] Furthermore, in the context of the kit according to the invention, the term "in combination with" includes that one or the other of the two components may be administered before, after, and / or simultaneously with the other component (optionally repeatedly). When used in this context, the terms "simultaneous administration" and "simultaneous administration with" include that individual amounts / doses of the relevant compound of the invention and other pharmaceutically active ingredients are administered within 48 hours (e.g., 24 hours) of each other.

[0412] Regarding the above-mentioned combination formulations and kits, it is preferred that the other active pharmaceutical ingredient is an anti-inflammatory agent, or an agent known to cause inflammation as a side effect, as described above.

[0413] Wherever the word “about” is used herein (e.g., in the context of amounts such as the concentration and / or dosage, molecular weight, or pH of the active ingredient and / or the compounds of the invention), it should be appreciated that such variables are approximate and can therefore vary by ±10% relative to the numbers specified herein, for example ±5%, preferably ±2% (e.g., ±1%). In this respect, the term “about 10%” means, for example, ±10% with respect to the number 10, i.e., between 9% and 11%.

[0414] The advantages of the compounds of this invention lie in their diverse applications, including:

[0415] • As a bioactive agent, it is used for a variety of conditions characterized by inflammation, whether the condition is an inflammatory disease of an organ as described above, or is related to or characterized by inflammation (e.g., wounds or burns), and / or for surgical and / or cosmetic applications.

[0416] • In combination with active pharmaceutical ingredients, in combination therapy, or as part of the following by exhibiting a more inert function:

[0417] Pharmaceutically acceptable excipients (e.g., adjuvants, diluents, or carriers).

[0418] o Medical devices, and / or

[0419] The medical device component of a drug-medical device combination.

[0420] The compounds, uses, and methods described herein may also have the following advantages: in the treatment of the conditions mentioned above, they may be more convenient and effective for physicians and / or patients, less toxic, have a wider range of activity, be more potent, produce fewer side effects, or have other useful pharmacological properties, whether for the treatment of inflammation, inflammatory disorders, or disorders characterized by inflammation as a symptom (including wounds) or otherwise.

[0421] The present invention is illustrated by the following embodiments, wherein... Figure 1 The proportion of unhealed wounds in a mouse model of acute wounds is shown; Figure 2 The amount of VEGF measured in wound tissue of a diabetic wound mouse model is shown. Figure 3 The measured amount of TGFβ1 is shown; Figure 4 This demonstrates the effect on swelling induced by acute inflammation in a mouse ear swelling model; Figure 5 and 6 The levels of Evans blue in rectal and anal tissues are shown, indicating the vascular permeability of the test compound; Figure 7 , 8 Figures 9 and 1 show the effects on body weight, ulcer surface area, and overall appearance in a TNBS-induced ulcerative proctitis model, respectively. Figure 10 These are photographic evidence of the effects of testing compounds on surgical wound healing; Figure 11 The bioadhesion properties of the compounds of the present invention are shown; and Figure 12 and Figure 13 The plasma concentrations of mesalazine and montelukast versus time are shown, respectively, when administered with or without the compounds of the present invention. Example

[0422] Example 1

[0423] (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2 (SEQ ID No:40)

[0424] Fmoc-Lys(Boc)-Wang resin (9.15 g, GLS180322-41301, GL Biochem, Shanghai, China) was loaded into a glass reaction column.

[0425] Dichloromethane (DCM, 200 mL; Shandong Jinling Chemical Industry Co., Ltd., Shandong, China) was added to the column and the resin was allowed to soak for about half an hour. The DCM was then removed by vacuum filtration.

[0426] The resin was washed three times with N,N-dimethylformamide (DMF, 200 mL; Shandong Shitaifeng Fertilizer Industry Co. Ltd., Shandong, China).

[0427] A 20% piperidine solution (200 mL; Shandong Shitaifeng Fertilizer Industry Co. Ltd., Shandong, China) was added to DMF as a deprotection solution and the reaction was carried out for 20 minutes. The solution was then removed by vacuum filtration and the column was washed six times with DMF.

[0428] Fmoc-DOPA (acetone condensate)-OH (4.14 g; GLS190219-21003, GL Biochem, Shanghai, China) and 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethylamineonium tetrafluoroborate (TBTU, 2.89 g; GLS170805-00705, GLBiochem, 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). After reacting for 30 minutes, a Kaiser test was performed with a small amount of resin; the yellow color of the solution and the appearance of a colorless gel indicated the completion of the reaction. The solvent was removed by vacuum filtration.

[0429] Repeat the above coupling steps to couple the same amount (in mol) of the remaining amino acids: 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.

[0430] After Fmoc-Ala-OH is coupled to the resin, the coupling steps are repeated starting with Fmoc-Lys(Boc)-OH and then with Fmoc-DOPA (acetone condensation)-OH, 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.

[0431] In a separate procedure, after coupling Fmoc-Ala-OH to the resin, a deprotection step was performed to remove the Fmoc protection on Dopa. The resin was washed three times with DMF (200 mL each time). A 20% piperidine solution (200 mL) in DMF was added as the deprotection solution and reacted for 20 minutes. The resin was then washed three times each with the following solvents: DMF (200 mL each time), DCM (200 mL each time), and methanol (200 mL each time; Xilong Scientific Co., Ltd., Guangdong, China). The resin was dried under vacuum for approximately 2 hours.

[0432] 160.0 mL (i.e., 10 mL / g dry resin) of lysis buffer was added to submerge the peptide-containing compound bound to the resin. The lysis buffer consisted of 95% trifluoroacetic acid (TFA), 2.5% water, and 2.5% triisopropylsilane (Tis). After lysis for approximately 2 hours, the solid support was removed by filtration, and the filtrate was collected under reduced pressure. The filtrate was precipitated with 1600 mL (i.e., 10 mL / ml filtrate) of diethyl ether (Xilong Scientific Co., Ltd., Guangdong, China), and the precipitate was collected by filtration. The precipitate was dried under vacuum for approximately 2 hours, yielding 7.53 g of the crude title compound.

[0433] The crude product was first analyzed in pure water as a 1 mg / mL sample and detected using a Shimadzu LCMS-8050 system. The analytical column was an Agilent ZORBAX Eclipse SB-C18 (4.6 × 250 mm, 5 μm column; detection: UV at 220 nm; solvent A: 0.1% TFA in MeCN, solvent B: 0.1% TFA in water, linear gradient from 5% to 90% of solvent A concentration over 50 minutes; flow rate: 1.0 mL / min; sample volume: 10 μL).

[0434] The target peak was eluted at 11.926 min and had the expected molecular weight (MS: m / z 2380.6) with a purity of 60.345%.

[0435] Then, 7.5 g of the crude product was dissolved in 80 mL of pure water and purified using an LC3000 semi-precipitation apparatus. The preparative column model was a Dubhe-C18 model (Hanbon Sci.&Tech.Co.,Ltd., Jiangsu, China) (50*250 mm). Column; Detection: UV at 220 nm). Calculate the appropriate elution gradient from the LCMS detection procedure (solvent A: 0.1% TFA in MeCN, solvent B: 0.1% TFA in water, linear gradient from 5% to 20% solvent A concentration over 30 minutes; flow rate 60.0 mL / min). Collect fractions and analyze using a Shimadzu LC-20 HPLC system (column as described above, except the linear gradient is from 5% to 30% solvent A concentration over 25 minutes).

[0436] The fractions with a purity of 98% were then mixed for anion exchange. This was performed using an LC3000 semi-preparative apparatus (preparative column model: Dubhe-C18, as described above). The fractions were diluted once with pure water and loaded directly onto the column, followed by washing the column with 0.37% ammonium acetate in pure water for approximately 20 minutes at a flow rate of 60 mL / min, then washing with pure water for another 20 minutes, and then eluting with the following gradient (solvent A: 0.1% HAc in MeCN, solvent B: 0.1% HAc in water, linear gradient from 5% to 20% of solvent A concentration over 30 minutes; flow rate 60.0 mL / min). The fractions were collected and analyzed using a Shimadzu LC-20 HPLC system (column and conditions as described above). The fractions with a purity of 98% were mixed and lyophilized to give 3.06 g of purified title compound.

[0437] Example 2

[0438] (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys) 3- 5 (SEQ ID No: 41, 42 and 43)

[0439] Repeat one or more procedures as described in Example 1. Once the amino acid is coupled to the resin, repeat the procedure as described in Example 1 three more times to provide (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)5 (SEQ ID No:43).

[0440] Five decapeptide repeat product peaks were detected by LCMS at 13.511 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; gradient: 0–25 min, 5%–30% B; flow rate: 1.0 mL / min; volume: 10 μL), and the compounds were separated.

[0441] MS (products of five decapeptide repeat sequences): m / z 5924.6

[0442] To provide (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)3 (SEQ ID No:41) and (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)4 (SEQ ID No:42), the procedure as described in Example 1 may be repeated once or twice if necessary.

[0443] MS (products of three decapeptide repeat sequences): m / z is 3562.0

[0444] MS (products of four decapeptide repeat sequences): m / z is 4743.3

[0445] Example 3

[0446] (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2-Lys(SEQ ID No:45)

[0447] Fmoc-Lys(fmoc)-Wang resin (9.9 g, GLS191010-41303, GL Biochem, Shanghai, China) was loaded into a glass reaction column.

[0448] The method described above is the same as the first method described in Example 1, except that Fmoc-Lys(Boc)-OH is first coupled to the resin, and then Fmoc-Dopa(acetone)-OH, 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 are coupled. In addition, the amounts of amino acids, TBTU and DIPEA are twice that of Example 1 (in moles).

[0449] Repeating the essentially same method yielded another batch of crude title compounds (yield 7.89 g). Analysis showed the target peak was eluted at 11.376 min, with the expected molecular weight (MS: m / z 2508.8). Purity was 68.985%.

[0450] The 7.8 g crude product was then purified as described in Example 1 above, and freeze-dried to give 2.57 g of pure title compound.

[0451] Example 4

[0452] (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2-Orn or -Dap (SEQ ID No:46 and 47)

[0453] The method is the same as that described in Example 3, except that the resin used is Fmoc-Orn(fmoc)-Wang resin or Fmoc-Dap(fmoc)-Wang resin.

[0454] Example 5

[0455] [(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2-Lys]2-Lys(SEQ ID No:48)

[0456] The method is the same as that described in Example 3, starting with Fmoc-Lys(fmoc)-OH, followed by Fmoc-Lys(boc)-OH, Fmoc-Dopa (acetone condensation)-OH, 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, and the amounts of amino acids, TBTU, and DIPEA are twice (in moles) compared to Example 3.

[0457] Repeating the essentially same method yielded another batch of crude title compounds (yield 15.29 g). Analysis showed the target peak was eluted at 11.563 min, with the expected molecular weight (MS: m / z 5127.62). Purity was 52.126%.

[0458] The 15.2 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.96 g of pure title compound.

[0459] Example 6

[0460] [(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2-Orn]2-Orn or Dap]2-Dap(SEQ ID No: 49 and 50)

[0461] The method described is the same as that in Example 5, except that Fmoc-Orn(fmoc)-Wang resin or Fmoc-Dap(fmoc)-Wang resin is used instead. The first amino acid coupled to the resin is Fmoc-Orn(fmoc)-OH or Fmoc-Dap(fmoc)-OH (as the case may be), instead of Fmoc-Lys(fmoc)-OH.

[0462] Example 7

[0463] {[(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2-Lys]2-Lys}2-Lys(SEQ ID No:51)

[0464] The method is the same as that described in Example 5, except that the first amino acid coupled to the resin is Fmoc-Lys(fmoc)-OH, followed by Fmoc-Lys(fmoc)-OH, and then Fmoc-Lys(boc)-OH, Fmoc-DOPA (acetone condensation)-OH, 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, and the amounts of amino acids, TBTU and DIPEA are twice that of Example 5 (in moles).

[0465] Repeating the substantially same method yielded another batch of crude title compounds (yield 28.14 g). Analysis showed the target peak was eluted at 11.753 min, with the expected molecular weight (MS: m / z 10365.2). Purity was 30.423%.

[0466] The 28.1 g crude product was then purified as described in Example 1 above, and freeze-dried to give 5.72 g of pure title compound.

[0467] The compound of Example 7 is referred to as "Compound B" below.

[0468] Example 8

[0469] {[(Dopa-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys)2-Lys]2-Lys}2-Lys(SEQ ID No:52)

[0470] The method is the same as that described in Example 7, except that the fourth amino acid coupled to the resin is Fmoc-Tyr(tBu)-OH instead of Fmoc-DOPA (acetone condensation)-OH, and after Fmoc-Ala-OH is coupled to the resin, another amino acid, Fmoc-DOPA (acetone condensation)-OH, is coupled to the resin.

[0471] MS: m / z 11671.1

[0472] The compound of Example 8 is referred to as "Compound A" below.

[0473] Example 9

[0474] {[(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2-Orn or Dap]2-Orn or Dap}2- Orn or Dap (SEQ ID No: 53 and 54)

[0475] The method is the same as that described in Example 7, except that Fmoc-Orn(fmoc)-Wang resin or Fmoc-Dap(fmoc)-Wang resin is used instead. The first two amino acids coupled to the resin are Fmoc-Orn(fmoc)-OH or Fmoc-Dap(fmoc)-OH (whichever is appropriate), instead of Fmoc-Lys(fmoc)-OH.

[0476] Example 10

[0477] (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2 (SEQ ID No:40) self-crosslinking product mixture

[0478] 4 mg of the product from Example 1 (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2 (SEQ ID No: 40) and 0.33 mg of mushroom tyrosinase (Sigma: T3824-250 KU, 2687 units / mg) were added to 2.2 mL of phosphate buffer solution (100 mM, pH 6.5) containing 25 mM ascorbic acid. The mixture was stirred for 2 hours. Then, 0.15 mL of 1 M HCl solution was added to the mixture to stop the reaction.

[0479] Samples were taken for MALDI-TOF mass spectrometry analysis. The results revealed that the molecular weight of the linear peptide (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys)2 with two repeating sequences could be increased by 2 to 6 times compared to its original molecular weight.

[0480] Example 11

[0481] Synthesis of other bifurcated branched peptides

[0482] The following peptides were synthesized using essentially the same methods as those described in Example 3 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence:

[0483] (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys)2-Lys (SEQ ID No: 55);

[0484] (Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys (SEQ ID No: 56);

[0485] (Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys (SEQ ID No: 57);

[0486] (DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys (SEQ ID No: 58);

[0487] (Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys(SEQ ID No:59); and

[0488] (DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys (SEQ ID No: 60).

[0489] The crude yields and purities, retention times, MS values, and final yields of these peptides are shown in Table 1 below.

[0490] Table 1

[0491]

[0492]

[0493] Example 12

[0494] Synthesis of other tetrabranched peptides

[0495] The following peptides were synthesized using essentially the same methods as those described in Example 5 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence:

[0496] [(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys)2-Lys]2-Lys (SEQ ID No: 61);

[0497] [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 62);

[0498] [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 63);

[0499] [(DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 64);

[0500] [(Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 65); hereinafter referred to as "compound C"); and

[0501] [(DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 66).

[0502] The crude yields and purities, retention times, MS values, and final yields of these peptides are shown in Table 2 below.

[0503] Table 2

[0504]

[0505] Example 13

[0506] Synthesis of other octapeptides

[0507] The following peptides were synthesized using essentially the same methods as those described in Example 7 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence:

[0508] {[(Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys)2-Lys]2-Lys}2-Lys (SEQ IDNo:67);

[0509] {[(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys}2-Lys (SEQ IDNo:68);

[0510] {[(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys}2-Lys(SEQ IDNo:69);

[0511] {[(DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys}2-Lys(SEQID No:70);

[0512] {[(Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys}2-Lys (SEQ IDNo:71); and

[0513] {[(DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys}2-Lys (SEQ ID No: 72).

[0514] The crude yields and purities, retention times, MS values, and final yields of these peptides are shown in Table 3 below.

[0515] Table 3

[0516]

[0517] Example 14

[0518] (HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys(SEQ ID No:73)

[0519] The title compound was prepared using essentially the same method as in Example 3 above, except that it was finally coupled with 3,4-dihydroxyhydrocinnamic acid (3.28 g, Macklin, Shanghai, China) to obtain 7.78 g of crude title compound.

[0520] Analysis showed that the target peak was eluted at 10.684 minutes, possessing the expected molecular weight (MS: m / z 2805.0). The purity was 62.283%.

[0521] The 7.7 g crude product was then purified as described in Example 1 above, and freeze-dried to give 2.46 g of pure title compound.

[0522] Example 15

[0523] [(HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys(SEQ ID No: 74)

[0524] The title compound was prepared using essentially the same method as in Example 5 above, except that it was finally coupled with 3,4-dihydroxyhydrocinnamic acid (6.56 g) to obtain 15.77 g of crude title compound.

[0525] Analysis showed that the target peak was eluted at 10.727 min, with the expected molecular weight (MS: m / z 5720.1). The purity was 48.274%.

[0526] The 15.7 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.59 g of pure title compound.

[0527] Example 16

[0528] {[(HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys}2-Lys(SEQ ID No:75)

[0529] The title compound was prepared using essentially the same method as in Example 7 above, except that it was finally coupled with 13.12 g of 3,4-dihydroxyhydrocinnamic acid to obtain 28.69 g of crude title compound.

[0530] Analysis showed that the target peak was eluted at 10.833 min, with the expected molecular weight (MS: m / z 11551.1). The purity was 28.821%.

[0531] The 28.6 g crude product was then purified as described in Example 1 above, and freeze-dried to give 5.62 g of pure title compound.

[0532] Example 17

[0533] [(Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys)2-Lys]2-Lys(SEQ ID No:76)

[0534] The title compound was prepared using essentially the same method as described in Example 5 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 14.97 g of crude title compound.

[0535] Analysis showed that the target peak was eluted at 11.578 minutes, possessing the expected molecular weight (MS: m / z 5191.2). The purity was 52.553%.

[0536] The 14.9 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.87 g of pure title compound.

[0537] Example 18

[0538] [(HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys)2-Lys]2-Lys(SEQ ID No: 77)

[0539] The title compound was prepared using essentially the same method as described in Example 15 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 15.66 g of crude title compound.

[0540] Analysis showed that the target peak was eluted at 10.697 minutes, possessing the expected molecular weight (MS: m / z 5720.0). The purity was 49.033%.

[0541] The 15.6 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.62 g of pure title compound.

[0542] Example 19

[0543] (HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys)2-Lys(SEQ ID No:78)

[0544] The title compound was prepared using essentially the same method as described in Example 14 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 7.83 g of crude title compound.

[0545] Analysis showed that the target peak was eluted at 10.594 minutes, possessing the expected molecular weight (MS: m / z 2869.1). The purity was 61.036%.

[0546] The 7.8 g crude product was then purified as described in Example 1 above, and freeze-dried to give 2.51 g of pure title compound.

[0547] Example 20

[0548] [(DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys)2-Lys]2-Lys(SEQ ID No:79)

[0549] The title compound was prepared using essentially the same method as described in Example 5 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 14.67 g of crude title compound.

[0550] Analysis showed that the target peak was eluted at 11.554 minutes, with the expected molecular weight (MS: m / z 5191.3). The purity was 50.576%.

[0551] The 14.6 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.64 g of pure title compound.

[0552] Example 21

[0553] (DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys)2-Lys(SEQ ID No:80)

[0554] The title compound was prepared using essentially the same method as described in Example 3 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 7.75 g of crude title compound.

[0555] Analysis showed that the target peak was eluted at 11.059 minutes, possessing the expected molecular weight (MS: m / z 2540.6). The purity was 65.384%.

[0556] The 7.7 g crude product was then purified as described in Example 1 above, and freeze-dried to give 2.36 g of pure title compound.

[0557] Example 22

[0558] (HCA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys)2-Lys(SEQ ID No:81)

[0559] The title compound was prepared using essentially the same method as described in Example 14 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 7.45 g of crude title compound.

[0560] Analysis showed that the target peak was eluted at 10.489 minutes, possessing the expected molecular weight (MS: m / z 2446.1). The purity was 65.457%.

[0561] The 7.3 g crude product was then purified as described in Example 1 above, and freeze-dried to give 2.27 g of pure title compound.

[0562] Example 23

[0563] [(HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys)2-Lys]2-Lys(SEQ ID No:82)

[0564] The title compound was prepared using essentially the same method as described in Example 15 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 14.79 g of crude title compound.

[0565] Analysis showed that the target peak was eluted at 11.235 minutes, possessing the expected molecular weight (MS: m / z 5067.5). The purity was 53.853%.

[0566] The 14.7 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.37 g of pure title compound.

[0567] Example 24

[0568] [(Lys-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr)2-Lys] 2- Lys (SEQ ID No:83)

[0569] The title compound was prepared using essentially the same method as described in Example 5 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 14.26 g of crude title compound.

[0570] Analysis showed that the target peak was eluted at 11.478 min, possessing the expected molecular weight (MS: m / z 5063.3). The purity was 49.642%.

[0571] The 14.2 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.33 g of pure title compound.

[0572] Example 25

[0573] (Lys-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA)2-Lys(SEQ ID No:84)

[0574] The title compound was prepared using essentially the same method as described in Example 3 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 7.37 g of crude title compound.

[0575] Analysis showed that the target peak was eluted at 10.672 min, possessing the expected molecular weight (MS: m / z 2540.2). The purity was 61.252%.

[0576] The 7.3 g crude product was then purified as described in Example 1 above, and freeze-dried to give 2.28 g of pure title compound.

[0577] Example 26

[0578] [(Lys-Ala-Lys-Hyp-Ser-DOPA-Hyp-Hyp-Thr-DOPA)2-Lys]2-Lys(SEQ ID No:85)

[0579] The title compound was prepared using essentially the same method as described in Example 5 above, except that appropriate amino acids were used in the appropriate peptide coupling sequence to obtain 14.06 g of crude title compound.

[0580] Analysis showed that the target peak was eluted at 11.223 minutes, possessing the expected molecular weight (MS: m / z 5255.3). The purity was 50.577%.

[0581] The 14.0 g crude product was then purified as described in Example 1 above, and freeze-dried to give 4.25 g of pure title compound.

[0582] Example 27

[0583] Airbag model

[0584] Healthy adult male C57BL / 6 mice weighing between 20g and 30g were provided by Changzhou Cvens Experimental Animal Co., Ltd. Prior to any experiments, the mice were held in captivity under standard conditions (constant temperature or 22℃±2℃, with a 12-hour light-dark cycle) and fed standard mouse diet and water for approximately one week. The mice were randomly divided into 9 groups, as shown in Table 4, with 7 mice in each group.

[0585] General anesthesia was induced using 3% chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China; 1 mL / 10 g body weight) delivered intraperitoneally. The hair on the entire back was shaved and removed the day before the sterile air injection.

[0586] An air sac was created in mice by subcutaneous injection of sterile air (5 mL) into the intrascapular region. Three days later, air was injected again (3 mL) to maintain the sac. To induce acute inflammation, three days after the second injection, the animals were given an injection of sterile carrageenan solution (CP Kelco, Taixing, Jiangsu, China; 1%, 0.5 mL; produced by adding 0.1 g of carrageenan powder to a beaker containing 10 mL of 0.9% saline solution and stirring). Mice were pretreated with the test sample or medium 1 hour before and 23 hours after the carrageenan injection into the subcutaneous air sac. Animals were sacrificed 24 hours after the carrageenan injection.

[0587] Skin biopsies were performed from an air-filled bag. A portion of the biopsy was fixed in formalin (produced by adding ultrapure water to a total volume of up to 500 mL of 40% formaldehyde solution (Nanchang Rain Dew Experimental Equipment Co., Ltd., Nanchang, Hubei Province, China)) and analyzed by histological embedding, sectioning, and staining in solid paraffin.

[0588] Histological specimens were analyzed, and inflammation and edema scores were estimated. Inflammation scores were estimated by observing hematoxylin- and eosin (HE)-stained sections under an optical microscope. Scores were assigned based on the perceived level of inflammation (ranging from 1 to 3) (e.g., 1 point (mild) for a small number of inflammatory cells scattered throughout the area; 2 points (moderate) for a large number of inflammatory cells observed; and 3 points (severe) for diffuse infiltration). A similar scoring system was used for the level of edema after overall observation (3 points for most severe and 1 point for mild). Scores for each group are presented in Table 5.

[0589] After conducting some preliminary experiments to validate the model, experiments were carried out in which mice were treated by administering test samples or mediators according to Table 4 below. Compound A and Compound B were dissolved in saline solution at the concentrations indicated in Table 4 below (L = low dose, M = medium dose, H = high dose).

[0590] Compound A and Compound B were synthesized as described in Examples 8 and 7 above, respectively. Before use, the peptide powder was stored at -20°C. Dexamethasone (Dex) was obtained from Shanghai Aladdin Bio-Chem Technology Co., Ltd., Shanghai, China.

[0591] Table 4

[0592] Group deal with Dosage (μg / mouse) Comparison Untreated mice / Model Model + Salt Water / Dex Model + Dexamethasone 400 AL Model + Compound A 50 AM Model + Compound A 250 AH Model + Compound A 1250 BL Model + Compound B 50 BM Model + Compound B 250 BH Model + Compound B 1250

[0593] The histological specimens were analyzed and scored as outlined above, and the results are shown in Table 5 below.

[0594] Table 5

[0595] Group Inflammation score Edema score Total score Comparison 0.00 1.14 1.14 Model 2.30 2.71 5 Dex 0.57 1.57 2.14 AL 1.86 1.71 3.57 AM 1.57 1.57 3.14 AH 1.27 1.71 2.98 BL 1.83 2.00 3.83 BM 1.80 2.14 3.94 BH 1.56 1.57 3.13

[0596] Histological analysis showed that compounds A and B had certain anti-inflammatory effects compared with the model group.

[0597] Example 28

[0598] Acute wound model

[0599] Six- to eight-week-old male C57BL / 6 mice were provided by Changzhou Cvens Experimental Animal Co., Ltd. (Changzhou, Jiangsu Province, China). Before any experiments, the mice were kept in captivity under standard conditions (constant temperature of 22℃±2℃, with a 12-hour light-dark cycle) and fed standard mouse diet and water for approximately one week.

[0600] General anesthesia was induced by intraperitoneal administration of 3% chloral hydrate (1 mL / 10 g body weight). Hair on the back was shaved with an infant razor and depilated with cream. The skin area was wiped clean and sterilized twice with 75% alcohol.

[0601] Two circular incisions were made along the midline of the back using a 12mm EMS skin biopsy piercing instrument (Electron Microscopy Sciences, POBox 550, Hatfield Industrial Road 1560, PA 19440). These two incisions were tangent to each other, and the skin between the circles was cut along the superior and inferior tangent lines. The incisions were trimmed with scissors. The full thickness of the skin was removed to reach the fascia. The incisions were oval-shaped and left open without sutures.

[0602] The following different medications were applied topically to the wound once daily from day 0 to day 7 (see Table 6 below). The control group had no wounds. The model group received the same amount of saline. Each group consisted of 10 mice, except for the control group which had 5 mice.

[0603] 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 physiological saline to prepare a solution with a concentration of 5000 IU / mL. The dose of rhEGF used in this experiment was 1285 IU / wound. Compound A and compound B were dissolved in saline at the concentrations indicated in Table 6 (L = low dose, M = medium dose, H = high dose).

[0604] Table 6

[0605] Group Medication dosage (μg / wound / day) Comparison / Model / EGF <![CDATA[1285IU(500IU / cm 2 )]]> AL 1.65 AM 16.45 AH 164.5 BL 1.65 BM 16.45 BH 164.5

[0606] Photographs of each wound were taken every other day, starting from day 0. The photographs were scanned into a computer, and the wound area was calculated using ImageJ image analysis software (National Institutes of Health).

[0607] The area of ​​unhealed wounds is expressed as a percentage of the original wound area:

[0608] A t / A0×100%,

[0609] Where A0 and A t These refer to the initial area on day 0 and the wound area on the measurement date (time t), respectively.

[0610] The rate of unhealed wounds is Figure 1 As shown in the figure, all wounds in the test group healed faster than those in the model group. Compounds A and B appeared to significantly improve wound healing, especially at medium concentrations of compound A and high concentrations of compound B.

[0611] Example 29

[0612] Diabetic wound model

[0613] Male db / db mice (C57BL / KsJ-db / db, weighing 35-45g / mouse) aged 8 to 12 weeks were provided by Changzhou Cvens Experimental Animal Co., Ltd. Prior to any experiments, the mice were kept in captivity under standard conditions (constant temperature of 22℃±2℃, with a 12-hour light-dark cycle) and fed standard mouse diet and water for approximately one week.

[0614] General anesthesia was induced using 3% chloral hydrate (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China; 1 mL / 10 g body weight) intraperitoneally. Hair on the back was shaved with an infant razor and then treated with depilatory cream. The skin area was wiped clean and sterilized twice with 75% alcohol.

[0615] A circular incision was made on the back using an 18mm diameter EMS skin biopsy puncturer. Full-thickness skin was removed, and the incision depth reached the fascia. The incision was left open without sutures.

[0616] Different medications were applied topically to the wound once daily from day 0 to day 18, at a dose of 50 μL / wound, as shown in Table 7 below. The control group had no wounds.

[0617] The same amount of saline was given to the model group. Each group consisted of 12 mice, except for the control group which had 8 mice. For the control group, skin fragments removed during wound formation were used as samples on day 7.

[0618] Purchase and prepare recombinant human epidermal growth factor (rhEGF) according to the manufacturer's instructions. Dissolve lyophilized rhEGF powder (100,000 IU / vial) in 20 mL of physiological saline to prepare a solution with a concentration of 5,000 IU / mL. The working dose of rhEGF used in this experiment was 1285 IU / wound.

[0619] Dissolve compounds A and B in saline solution at the concentrations indicated in Table 7 below (L = low dose, M = medium dose, H = high dose). Apply 50 μL of each solution to the wound surface daily.

[0620] Table 7

[0621] Group Medication dosage (μg / wound / day) Comparison <![CDATA[ / ]]> Model physiological saline EGF 1285IU AL 1.65μg AM 16.45μg AH 164.5μg BL 1.65μg BM 16.45μg BH 164.5μg

[0622] Vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) play important roles in wound healing. VEGF and TGF-β1 are commonly co-expressed in tissues where angiogenesis occurs. The levels of these two factors in wound tissue were also examined, and their effects were shown. Figure 2 and 3 middle.

[0623] The results showed that at different times, the levels of VEGF and TGF-β1 in all test groups were higher than those in the model group, indicating that compounds A and B stimulated the production of VEGF and TGF-β1.

[0624] Example 30

[0625] Mouse ear swelling model

[0626] Thirty healthy male BALB / c mice, aged 6-8 weeks and with an average weight of 18-25g, were provided by Changzhou Cvens Experimental Animal Co., Ltd. and housed and cared for for approximately one week before the experiment. The temperature was maintained at approximately 25°C to 27°C, humidity at 74%, with a 12-hour light-dark cycle, and free access to food and water. The mice were randomly divided into 6 groups, as shown in Table 8, with 5 mice in each group. Compound A and Compound B were dissolved at the concentrations indicated in Table 8 below (L = low dose, M = medium dose, H = high dose).

[0627] Table 8

[0628] Group Drug concentration Model / Dex Cream 10μg / g AL 0.5mg / g AH 1mg / g BL 0.5mg / g BH 1mg / g

[0629] A hydrogel containing the amounts of peptides described in Table 8, as well as methylcellulose (2.5%), propylene glycol (11%), and glycerol (11%), was prepared, with the pH adjusted to 5.5 by adding acetic acid (pH adjuster; 0 to 0.5 g). All excipients were obtained from Sinopharm Chemical Reagent Co., Ltd. The gel was prepared with water for injection.

[0630] Dexamethasone acetate cream (Dex cream; 5 mg / 10 g (meaning 5 mg Dex is contained in 10 g of cream), Fuyuan Pharmaceutical Co. Ltd., Anhui, China) was used as a positive control.

[0631] The left ear of each mouse was used as a self-control. The right ear of each mouse was treated with the above-mentioned compounds at specified concentrations (Table 8).

[0632] Approximately 0.1 g of each gel and Dex cream was applied to the right ear (internal and external) of each group of mice. A blank gel matrix was applied to the ears of the model groups. One hour later, 20 μL of xylene (Shanghai Aladdin Bio-ChemTechnology Co., Ltd.) was applied to the same ear of each mouse.

[0633] Forty minutes after xylene administration, mice were euthanized by cervical dislocation. The left and right ears were removed. A piece of each ear was harvested from the same site using an 8mm diameter EMS skin biopsy bag. The weight was recorded, and the swelling percentage was calculated as a percentage using the following formula:

[0634] (Right ear weight - Left ear weight) / Left ear weight × 100

[0635] The results are shown in Table 9 and Figure 4 As shown in the image.

[0636] Table 9

[0637] Model DEX AL AH BL BH Swelling rate 79％ 35％ 63％ 43％ 43％ 38％ SD 0.166 0.198 0.184 0.231 0.095 0.17

[0638] The results showed that compounds A and B could significantly eliminate edema caused by inflammation.

[0639] Example 31

[0640] Preparation of membranes coated with compounds A and B

[0641] Cut 0.2 μm microfiltration membranes (Jinteng Corp., China) into 2.5 cm diameter discs and place them in three separate containers (10-15 discs per container). Add approximately 10 mL of 5 mg / mL stock solutions of compound A and compound B to each container, ensuring all membranes are completely submerged. While shaking, add PBS buffer (pH 8.0) dropwise to each container. Check the pH of the reaction mixture periodically until it reaches 7. Carefully seal the containers and shake continuously for 8 hours. Then pour off the reaction mixture and wash the membranes with 5 mL of PBS buffer each time until the wash eluent becomes colorless. Finally, place the membranes in a cool, dry place. Use the resulting membranes coated with compounds A and B for antioxidant testing.

[0642] Antioxidant capacity (AC) was measured using a modified DPPH (2,2-diphenyl-1-picrylhydrazine) method (as follows). Each membrane of equal weight was cut into small pieces and placed in separate 2 mL centrifuge tubes. A 0.1 mM solution of DPPH in methanol was prepared, and 600 μL of the DPPH solution was added to ensure complete submersion of all fragments. The tubes were then kept at room temperature in the dark for 3 hours, followed by centrifugation for 5 min. 300 μL of the supernatant was added to a 96-well plate, and measurements were taken at 517 nm using a microplate reader. The AC value for each membrane was calculated according to the formula shown below, where A0 is the absorbance of the DPPH solution alone, and A... m It is the absorbance of each membrane sample.

[0643] AC(%)=(A0-A m ) / A0×100%

[0644] The antioxidant properties of membranes coated with compounds A and B were tested on days 0 (D0), 3 (D3), 7 (D7), and 10 (D10) to examine stability. The results are shown in Table 10.

[0645] Table 10

[0646] sample D0 D3 D7 D10 Membrane coated with compound A 70.85％ 68.16％ 65.82％ 68.82％ Membrane coated with compound B 68.00％ 65.52％ 68.97％ 65.97％

[0647] The results showed that both compound A and compound B could be coated onto the membrane. The coated membrane exhibited antioxidant properties, which lasted for at least 10 days.

[0648] Example 32

[0649] Croton oil-induced rat anal swelling model I

[0650] A croton oil mixture was prepared by mixing one part distilled water, four parts pyridine (Nanjing Chemical Reagent Co., Ltd.), five parts ether (China Pharmaceutical Group Chemical Reagents Co., Ltd.), and ten parts 6% croton oil (Shanghai Yuanye Biotechnology Co., Ltd.) ether solution.

[0651] Six- to eight-week-old Sprague Dawley (SD) rats, half male and half female, with an average weight of 180-220g, were provided by Changzhou Cvens Experimental Animal Co., Ltd. (Changzhou, Jiangsu Province, China). Prior to any experiments, the rats were kept in captivity under standard conditions (constant temperature of 22℃ ± 2℃, with a 12-hour light-dark cycle) and fed a standard mouse diet and water for approximately one week.

[0652] Fifty-six rats were randomly divided into seven groups (as shown in Table 11 below), with eight rats in each group. Compound A, Compound B, and Ma Yinglong Hemorrhoid Ointment were dissolved at the concentrations indicated in Table 11 below (L = low dose, M = medium dose, H = high dose).

[0653] Table 11

[0654] Group deal with Drug concentration Volume (μL) Comparison physiological saline / 200 Model Blank Gel / 200 MY-L Ma Yinglong Hemorrhoid Ointment Commercially available ointments 200 AL Compound A 0.5mg / g 200 AH Compound A 1mg / g 200 BL Compound B 0.5mg / g 200 BH Compound B 1mg / g 200

[0655] Rats were anesthetized by inhalation of isoflurane (China Pharmaceutical Group Chemical Reagents Co., Ltd.). The skin around the anus was disinfected with a 75% alcohol swab. Then, 0.16 mL of croton oil mixture was slowly dripped onto a cotton swab and inserted 0.5 cm into the rat's anus. The rat was lifted to keep its head upward (holding this position for 10 seconds), then the cotton swab was removed, and the croton oil mixture was evenly applied to the surrounding skin. The same volume was given to the control group, but with olive oil.

[0656] One hour after modeling, each group of rats was treated according to Table 11. The positive control drug was Ma Yinglong Hemorrhoid Ointment (MaYinglong Pharmaceutical Group Co., Ltd.). Gels of compounds A and B were prepared as described in Example 14. The drugs were administered twice daily, once in the morning and once in the evening, for three consecutive days.

[0657] Draw 200 μL of the corresponding drug into a 1 mL syringe (without the needle). Insert the syringe into the rectal canal and push approximately 160 mL of the corresponding test substance 1.5 cm into the rectal canal. Apply the remaining test substance to the skin around the anus. Hold the skin around the anus firmly for 1 minute to prevent drug leakage.

[0658] On the morning of the fourth day, 30 minutes after drug administration (200 μL / 100 g), 1% Evans blue (EB) was injected into the tail vein. The rats were euthanized by cervical dislocation 30 minutes later.

[0659] The rat was placed in a supine position on a dissecting board, and its abdomen was opened. The rectal and anal tissue (15 mm in length) was separated and weighed, and EB dye present in the tissue was extracted using 1 mL of formamide.

[0660] Transfer all samples to a 55°C water bath or heating block. Incubate for 24 hours, then extract EB from the tissue. Centrifuge the formamide / EB mixture to precipitate any remaining tissue debris. Measure absorbance at 610 nm using 500 μL of formamide as a blank.

[0661] The content of EB in rectal and anal tissues was calculated using the extravasated EB amount (in ng / mg tissue) to assess vascular permeability. Results were... Figure 5 The study showed that the gels containing compounds A and B could reduce inflammatory swelling induced by croton oil application, as indicated by changes in EB content across different treatments. Decreased EB concentrations indicated vascular permeability.

[0662] Example 33

[0663] Radiation proctitis

[0664] A gel containing 0.5g of compound C was prepared (see Example 12 above), the gel also comprising the following components: methylcellulose (2.2g; Shandong Guangda Technology Development Co., Ltd., Shandong, China), glycerol (11g) and propylene glycol (11g, both from Sinopharm Chemical Reagent Co., Ltd.) and purified water (75.3g).

[0665] For a 1.5 mg / g gel, the amounts of compound C and water were adjusted accordingly.

[0666] Mix methylcellulose and water together and stir until a homogeneous colloidal suspension is formed. Then, add peptide powder, glycerin, and propylene glycol to the methylcellulose / water mixture and stir the resulting mixture rapidly for 5 minutes to obtain the final product.

[0667] Male Wistar rats weighing 180-220g were obtained from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. All animals were kept in standard cages with a 12-hour light-dark cycle, fed standard rodent diets and tap water.

[0668] Rats were anesthetized with an intraperitoneal injection of 10% chloral hydrate (3.3 mL / kg). The rats were restrained in a supine position by their tails and limbs and strapped to cardboard. Irradiation was delivered using an Elekta Synergy medical linear accelerator (Elekta Limited, UK). All animals, except the sham group, received a single, continuous pelvic irradiation. The distance from the animal to the source was 100 cm. The irradiated area was 2 cm × 5 cm, 5 cm upward from the anus. The dose was 17.5 Gy at a dose rate of 600 cGy / min.

[0669] After irradiation, the animals were returned to their cages to recover naturally. Animals in the sham surgery group (“sham”) were anesthetized intraperitoneally without irradiation. Daily food intake and body weight were measured in the rats, and overall observations were performed daily.

[0670] Day 1 (D1) was defined as the drug administration day, 24 hours after modeling. Rats in the sham surgery group and the model group (“Model”) were given a blank gel. Rats in the treatment group were given a rectal dressing containing the gel (300 μL / rat) at a high dose (1.5 mg / g; “CH”) or a low dose (0.5 mg / g; “CL”) once daily for 7 consecutive days (D1 to D7). To reduce intestinal motility and prolong the duration of gel presence in the rectum, all animals were given an intraperitoneal injection of 6 mL / kg of 5% chloral hydrate daily prior to administration. The drug was introduced into the rectum approximately 6 cm via an intragastric needle. Sampling was performed on day 8 (D8). Rats were fasted for at least 12 hours prior to administration.

[0671] Rats were anesthetized by intraperitoneal injection of chloral hydrate and euthanized by exsanguination via the posterior artery. The colon and rectum, approximately 7 cm long, were separated about 0.3 cm from the edge of the perianal hairline. The specimens were trimmed, and 1 cm samples of the proximal and distal colon and rectum were removed separately by the same person.

[0672] The intestinal tract was then dissected longitudinally, photographed, and weighed. The samples were fixed in 10% formaldehyde solution for 48 hours and stained with hematoxylin and eosin (HE), and then examined by a pathologist (unaware of the study) using an optical microscope.

[0673] Each sample was graded as follows: 0 = normal or minor changes that could not be (definitely) attributed to radiation; 1 = minor radiation damage (mild inflammation and / or minor crypt changes); 2 = mild damage (more significant inflammation and / or crypt damage); 3 = moderate damage (must have significant epithelial loss, with variable degree of inflammation); and 4 = severe damage (ulceration, necrosis).

[0674] The rate of weight gain at day 8 (defined as: body weight at day 8 - initial body weight) / initial body weight x 100%) is shown in Table 12 below. A higher rate indicates better physiological condition. Compound C was dissolved at the concentrations indicated in Table 12 below (L = low dose, H = high dose).

[0675] Table 12

[0676] Group dose average value(%) Standard deviation False / 5.7030 3.7042 Model / -1.7047 5.2056 CL 0.5mg / g 0.8076 5.5920 CH 1.5mg / g 3.7538 4.0502

[0677] The results showed that the gel containing compound C prevented weight loss caused by radiation proctitis in a dose-dependent manner.

[0678] Example 34

[0679] Radiation vaginitis

[0680] A 45-year-old female patient diagnosed with cervical cancer was treated with radiation therapy. Radiation was delivered using high-energy 6-12MV X-rays. The radiation dose was 1.8-2.0 Gy, administered five times a week. The radiation therapy was planned to be completed within four weeks.

[0681] Two weeks after her first irradiation, she began to experience pain and eventually developed vaginal bleeding and ulcers. This was diagnosed as radiation vaginitis.

[0682] The patient began using a gel prepared as described in Example 33 above, at a dose of 1.5 mg / g x 3 g, packaged in a special applicator for vaginal use. She used it twice daily during her radiation therapy. After 3 days, her bleeding stopped and her pain subsided. After her radiation therapy was completed, she continued using the gel for another two weeks. Her doctor examined her vagina and found that the ulcers had disappeared and there were no other signs of damage.

[0683] Example 35

[0684] Croton oil-induced rat anal swelling model II

[0685] In 50 rats randomly divided into 5 groups, the procedure was performed using compound C gel as described in Example 33 above, which was essentially the same as the procedure described in Example 32 above (with compound C at doses of 0.5 mg / g (low) and 1.5 mg / g (high) instead of the corresponding compound A and compound B gels).

[0686] As described in Example 32, the content of EB in rectal and anal tissue (8 mm in length) was calculated to assess vascular permeability, and the results were... Figure 6 The results are shown in the figure. These results demonstrate that compound C gel reduces inflammatory swelling induced by croton oil application in a dose-dependent manner.

[0687] Example 36

[0688] Ulcerative proctitis model

[0689] Fifty SD rats were randomly divided into five groups of ten rats each (sham surgery; model (blank gel; model); positive control (sulfasalazine, SSZ, 360 mg / kg; SSZ); compound C (1.5 mg / g dose (high)) and compound C (0.5 mg / g dose (low) (both prepared as described in Example 33 above)).

[0690] After a 24-hour fast, the animals were anesthetized with isoflurane. Except for the sham surgery group, the other four groups were anesthetized and administered 0.5 mL of 2,4,6-trinitrobenzenesulfonic acid (TNBS; 1 mL; Dalian Meilun Biotechnology Co., Ltd., China) solution in ethanol (6.05 mL; Shanghai Aladdin Biochemical Technology Co., Ltd., China) and 18.071 mL of sterile water for injection (i.e., 18 mg TNBS / rat) via rectal irrigation through a latex tubing.

[0691] The length of the flexible tube inserted into the rectum was approximately 8 cm. After removing the tube, the rats were held under isoflurane anesthesia for another 15 minutes (day 0 of modeling), and then the animals were returned to their cages. The sham-operated group was given the same volume of physiological saline.

[0692] On the second day after modeling (Day 1), following anesthesia (intraperitoneal injection of pentobarbital (35 mg / kg, 1.5%, 0.233 mL / 100 g)), the gel was administered rectally at a dose of 0.5 mL / rat (model group and compound C group). The anus was clamped for 1 hour after administration and then released. In the positive control group, sulfasalazine was administered to SSZ via oral tube feeding. This was repeated once daily for 7 consecutive days (Day 1 to Day 7). The sham surgery group received no treatment.

[0693] The rats were observed daily for their overall condition, disease activity index (DIA), and body weight. On the second day after final administration (day 8), the animals were dissected, the entire colon was removed, and the colonic contents were cleared. Body weight was measured, and the colon was then opened longitudinally to expose the ulcer surface. The ulcer surface was measured, its overall appearance was assessed, and photographs were taken.

[0694] The results are respectively in Figure 7 (weight), Figure 8 (ulcer surface) and Figure 9 As shown in (General Appearance), and it is demonstrated that compound C reduces the severity of TNBS-induced ulcerative proctitis in a dose-dependent manner and thus promotes ulcer healing.

[0695] Example 37

[0696] Experimental gastric ulcer model

[0697] This experiment used SPF-grade SD rats, with 10 rats in each group. According to the evaluation methods for gastric ulcer health foods published by the State Food and Drug Administration of China, compound B was observed to have a protective effect against acute gastric ulcers induced by anhydrous ethanol after 30 days of continuous tube feeding.

[0698] The different groups and dosages are shown in Table 13 below.

[0699] Table 1 3

[0700]

[0701]

[0702] Open the omeprazole enteric-coated capsules (20 mg / capsule; Heilongjiang Norgas Pharmaceutical Co., Ltd., China) and dissolve the powder in water to prepare a 2 mg / mL solution. Dissolve compound B (see Example 7 above) powder in water to prepare two solutions with concentrations of 0.25 mg / mL and 0.75 mg / mL. The administration volume is 2 mg / kg.

[0703] All drugs were administered orally via tube feeding as shown in Table 13, once daily for 30 consecutive days. Rats had free access to water. On day 30, rats were fasted for 24 hours after the last drug administration. On day 31, each rat in all experimental groups (except the control group) was given 1.0 mL of anhydrous ethanol via tube feeding.

[0704] One hour later, all rats were euthanized and dissected to expose the intact stomach, which was then ligated at the pylorus. A 10% formaldehyde solution was administered into the stomach via irrigation and the stomach was immobilized for 20 minutes. After immobilization, the stomach was dissected and cut along the larger bend. The stomach contents were flushed off the lining with saline to expose the gastric mucosa.

[0705] The length and width of gastric mucosal bleeding were measured under a stereomicroscope or visually using calipers. A score was assigned based on the assessment criteria shown in Table 14 below to evaluate the damage.

[0706] Table 14

[0707]

[0708] The results are shown in Table 15 below.

[0709] Table 15

[0710] Group Damage score Comparison 0 Model 28 BL 24 BH 18 Omeprazole 20

[0711] The results showed that compound B could reduce alcohol-induced gastric bleeding and thus protect the gastric mucosa.

[0712] Example 38

[0713] Radiation damage in the nasal cavity

[0714] A 50-year-old male patient was diagnosed with paranasal carcinoma and underwent radiation therapy. Two weeks after treatment, he developed inflammation of the nose and sinuses, and experienced nasal congestion or difficulty breathing. The mucus in his nasal cavity and sinuses became thick and dry.

[0715] The patient used the nasal spray every two hours during the day and then packaged it into a nasal spray bottle. The nasal spray contained compound B dissolved in water at a concentration of 0.5 mg / mL (see Example 7 above). After three days, his nose became clear and was no longer congested. He continued to use the nasal spray as his radiation therapy progressed, and his nasal discomfort did not worsen.

[0716] Example 39

[0717] Radiation-induced stomatitis

[0718] A 79-year-old male patient was diagnosed with oral cancer and underwent radiation therapy.

[0719] Three weeks later, oral mucositis developed into ulcers on the mucous membranes lining the mouth, throat, and esophagus. This was extremely painful, and the patient was unable to eat.

[0720] The patient then used a solution of compound B (0.5 mg / mL; prepared as described in Example 38 above) as a mouthwash as needed. Pain was significantly reduced almost immediately upon application, and this relief lasted for one or two hours. The patient's stomatitis did not worsen as his treatment continued.

[0721] Example 40

[0722] Postoperative pain and bleeding

[0723] A 54-year-old man underwent surgery to remove hyperplastic black tissue from his right leg. Figure 10 As shown in the left figure. Immediately after the surgery, he applied 0.5 mg / mL of compound B spray (as described in Example 38, but packaged in a normal spray bottle) directly to the wound.

[0724] Taken 1 hour after surgery Figure 10The image on the right shows the bleeding has stopped, and there is only mild pain. The wound was fully healed after a week. The doctor said that with standard care, patients typically need at least ten days to fully recover from this type of procedure.

[0725] Example 41

[0726] Itching relief

[0727] A 36-year-old woman had suffered from hemorrhoids for many years. She usually didn't experience pain or bleeding, but she did experience itching, which greatly bothered her. She also had constipation.

[0728] The patient began using compound C gel (0.5 mg / g x 3 g; see Example 33 above) packaged in a rectal applicator, which she used once daily before bedtime.

[0729] The next day, she felt the itching lessen, and the itching disappeared after a week. At the same time, the patient's constipation became less frequent.

[0730] Example 42

[0731] Ulcerative colitis

[0732] A 39-year-old woman was diagnosed with an acute flare-up of ulcerative colitis. She had to defecate more than 10 times a day and was exhibiting severe bleeding from her colon.

[0733] She was treated with oral 5-aminosalicylic acid (also known as mesalazine or mesalamine) for 3 days, but her symptoms did not change.

[0734] Subsequently, over the first two days, she was given the same compound C gel as described in Example 41 above, three times at the same dosage.

[0735] She had to urinate less frequently, to 3 to 4 times a day. She continued using the gel once a day for another 7 days, after which her symptoms disappeared.

[0736] Example 43

[0737] Bioadhesive

[0738] The test was conducted in rats. Two approximately 1 cm incisions were made on the left side of SD rats (under isoflurane inhalation anesthesia). The left incision was left untreated, while the right incision was covered with a pinch of compound C powder. Each incision was closed with forceps for 10 seconds. The rats were then returned to their cages. After approximately 20 minutes, the rats awoke and began to move around. The untreated wound reopened, but the treated wound remained closed. After 24 hours, both incisions had almost healed, and the treated wound was smoother than the untreated one. (Left image) Figure 11 The image in the middle was taken immediately after the surgery, while the image on the right was taken 24 hours later.

[0739] Example 44

[0740] Pre-crosslinked compound I of the present invention was prepared using glutaraldehyde.

[0741] At room temperature, 100 mg of any of the peptides in SEQ ID No: 45, 48, 51, 57, 58, 63, 64, 69, or 70 was reacted with 2-100 mL of 0.01-0.5 M buffers (such as 0.01 M sodium acetate (pH 3.0), 0.1 M sodium acetate (pH 5.0), 0.2 M sodium phosphate (pH 6.0), and 0.5 M sodium phosphate (pH 8.0)) containing 0.01%-5.0% glutaraldehyde for 1-300 minutes. At the end of the reaction, a certain amount of NaHSO3 (equivalent to 80% glutaraldehyde) was added to terminate the reaction. The formulation was then thoroughly dialyzed against water to give one or more of the corresponding title compounds.

[0742] Example 45

[0743] Pre-crosslinked compound II of the present invention was prepared using glutaraldehyde.

[0744] Following essentially the same method as described in Example 44 above, 100 mg of [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 63) was reacted with 10 mL of 0.1 M sodium acetate buffer (pH 5.0) containing 0.05% glutaraldehyde for 10 minutes at room temperature. The degree of crosslinking was determined by size exclusion chromatography (SE-HPLC).

[0745] Example 46

[0746] The pre-crosslinked compound I of the present invention was prepared using an amide formation method.

[0747] At room temperature, mix 100 mg of any of the peptides in SEQ ID No: 45, 48, 51, 57, 58, 63, 64, 69 or 70 with 2-100 mL of pure water or 0.01-0.5 M buffers of different pH values ​​(approximately 3.0 to 8.0) (such as 0.01 M sodium acetate (pH 4.0), 0.05 M MES buffer (pH 5.0), 0.1 M MES buffer (pH 6.0) and 0.5 M sodium phosphate (pH 7.0)) for 0.5-72 hours, said buffers containing 1-500 mg of each condensing agent (such as N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (EDC / NHS) or 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholine hydrochloride (DMTMM) or other water-soluble condensing agents). At the end of the reaction, the formulation was thoroughly dialyzed against water to remove DMTMM and one or more corresponding title compounds were given.

[0748] Example 47

[0749] The pre-crosslinked compound II of this invention was prepared using an amide formation method.

[0750] Using essentially the same method as described in Example 46 above, 100 mg of [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 63) was reacted with 10 mL of 0.05 M MES buffer (pH 5.5) containing 100 mg DMTMM for 6 hours at room temperature. The degree of crosslinking was determined by size exclusion chromatography (SE-HPLC).

[0751] Example 48

[0752] Pre-crosslinked compound I of the present invention was prepared using the DOPA oxidation method.

[0753] 100 mg of any of the peptides listed in SEQ ID No: 45, 48, 51, 57, 58, 63, 64, 69, or 70 was reacted with 2-100 mL of pure water or with 0.01-0.5 M buffers of varying pH values ​​(approximately 3.0 to 8.0), such as 0.01 M sodium acetate (pH 5.0), 0.05 M MES buffer (pH 5.0), 0.5 M sodium phosphate (pH 7.0), and 0.1 M Tris buffer (pH 8.0)) containing 1-500 mg of each oxidant (such as peroxide, periodate) or multiple phenolases (such as tyrosinase). At the end of the reaction, 0.5%-5% (v / v) of 1 M HCl solution was added to the mixture to stop the reaction, yielding the pre-crosslinked title compound.

[0754] Example 49

[0755] Pre-crosslinked compound II of the present invention was prepared using the DOPA oxidation method.

[0756] Using essentially the same method as described in Example 48 above, 100 mg of [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 63) was reacted with 10 mg of mushroom tyrosinase (Sigma: T3824-250 KU, 2687 units / mg) in 10 mL of phosphate buffer (100 mM, pH 6.5). The mixture was stirred for 2 hours. Then, 1.5 mL of 1 M HCl solution was added to the mixture to stop the reaction. Samples were taken for MALDI-TOF mass spectrometry analysis. The results revealed that the molecular weight of the product increased by 2 to 6 times compared to its original molecular weight.

[0757] Example 50

[0758] Effect of the compounds of this invention on the activity of human influenza A virus H1N1

[0759] Prepare serum-free 1640 medium (RPMI 1640 medium; GIBCO / BRL; Thermo Fisher Scientific China, Nanjing, China) according to the manufacturer's instructions. Before use, prepare a complete medium containing 10% serum by adding newborn calf serum (Zhejiang Tianhang Biotechnology Co., Ltd., Luoshe, China), or prepare a maintenance solution by adding 2% of the same serum.

[0760] 40 mg of compound C was dissolved in 2 mL of aqueous sodium chloride (in water for injection, Jiangsu Hengrui Medicine Co., Ltd., Jiangsu Province, China) to prepare a 20 mg / mL stock solution.

[0761] Add 0.05 mL of stock solution to 1.95 mL of complete culture medium to prepare a 500 μg / mL drug solution (use maintenance solution instead of complete culture medium in tests 3 and 4 below). Then prepare working solutions with concentrations of 250, 125, 62.5, 31.25, 15.625, 7.8125, 3.9063, 1.9531, and 0.9766 μg / mL by two dilutions.

[0762] Cytotoxicity of Compound C

[0763] Vero cells were seeded in 96-well plates and grown into a monolayer. 0.2 mL of compound C at different concentrations (as above) was added to each well. For each concentration, it was repeated in 3 wells. Solvent and normal cell culture were used as negative controls. Cells were cultured at 35°C (5% CO2) for 24 h. 10 μL of Cell Counting Kit-8 (CCK-8, Sigma) was added to each well, mixed thoroughly, and cultured at 37°C for 2 h. OD was detected by enzyme-linked immunosorbent assay (ELISA). 450 The absorbance value (optical density) was used. The cell viability of untreated cells was set to 100%, and the cytotoxicity rate was calculated. Cytotoxicity rate (%) = (average absorbance of untreated cells - average absorbance of drug-treated wells) / average absorbance of untreated cells × 100%. The median lethal concentration (LC50) of the test drug was calculated. The results showed that compound C had no cytotoxicity at the test concentration.

[0764] The effect of compound C on the cytopathic effect of H1N1 after direct action.

[0765] Vero cells were seeded into 24-well plates and set aside until 70%–80% confluence was achieved. The virus was mixed with different concentrations of the test drug. The final concentrations of compound C were 2 μg / mL, 4 μg / mL, 8 μg / mL, 16 μg / mL, 32 μg / mL, 64 μg / mL, and 128 μg / mL. 0.1% SDS (manufactured by AMRESCO LLC, Robbins, Ohio, USA, and packaged by Bioharp Company, Hefei, China; purity: 99%) was used as a positive control and mixed with the virus for 1 hour. The cell culture medium was removed, and the cells were washed three times with PBS, then the virus / drug mixture was added and incubated for 1 hour. Untreated virus was used as a negative control. After 1 hour of incubation, the culture was washed three times with PBS and cultured continuously for 16–24 hours. RNA was extracted and quantitatively analyzed using a one-step QRT PCR kit to detect antiviral activity.

[0766] Calculate the viral inhibition rate. Set the cells in the non-drug-challenged group to 0%. Viral inhibition rate (%) = (1 - viral RNA% in the drug-treated group) × 100%. Calculate the IC50 at 50% effective concentration.

[0767] The results showed that the IC50 of compound C was >32 μg / mL in this test.

[0768] Example 51

[0769] [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 63) Effects of pre-administration on the pharmacokinetics of mesalazine

[0770] A solution of [(Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 63; hereinafter referred to as “sMAP”#) was prepared by weighing 22.5 mg and adding physiological saline to 15 mL to obtain a concentration of 1.5 mg / mL. Rats were treated by administering the test sample. Mesalazine suppositories were melted in a water bath at 40 °C to prepare a mesalazine suppository suspension.

[0771] Twelve SD rats (six males and six females) were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. and housed in a barrier facility for seven days. The temperature was maintained at 20°C–26°C, humidity at 40%–70%, with alternating light and dark cycles for 12 hours, and free access to food and water. As shown in Table 16 below, the rats were randomly divided into two groups of six (three males and three females) each.

[0772] Table 16

[0773] Group deal with dose level Dosage-volume ratio (per rat) Dosage route 1 Mesalazine 1g 200μL Rectal administration 2 Mesalazine + sMAP solution 1g 200μL Rectal administration

[0774] Rats were anesthetized by inhalation of 2% isoflurane. The rats were placed in a supine position, and the stool at the end of the rectum was squeezed out. The skin area around the anus was disinfected with 75% alcohol swabs.

[0775] The first group was given mesalazine suppository suspension directly, while the second group was given mesalazine suppository suspension 15 min after sMAP enema treatment (details of enema dosage and administration method are shown in Table 17 below).

[0776] Table 17

[0777]

[0778] Use a 2mL syringe to aspirate the air bag cavity to ensure there is no air left in the air bag after filling with water. Insert the catheter (2-WAY 8Ch / Fr 3-5mm) into the rat's anus to a depth of approximately 3cm (2cm from the end of the balloon; cut the tip of the catheter below the tip of the urethral catheter, and shorten it as much as possible while maintaining the integrity of the air cavity. Remove the guidewire and seal the urethral catheter with neutral silicone sealant, ensuring a smooth seal to minimize damage to the rectal mucosa). Using a 1mL syringe, draw the mesalazine suppository suspension onto a disposable feeding needle (soft needle) and insert it (approximately 3.5cm) into the anus. Slowly inject 200μL of the syringe contents into each rat.

[0779] While maintaining the perianal skin to secure the catheter and feeding needle, inject 1 mL of water into the air sac cavity using a 2 mL syringe to inflate it, and then quickly release the needle to observe for any gel leakage.

[0780] After injection, secure the catheter by wrapping it around the base of the tail with tape (lift the catheter approximately 3-5 cm from the anus, then cut it after connection and secure it with tape around the base of the tail). The pressure-sensitive tape should be applied based on the non-relaxation standard of the urinary catheter; the catheter should not be too tight and should not cause the animal to chew or feel uncomfortable. The catheter should be secured with tape twice: first, the catheter should be knotted, and second, the remaining catheter should be secured to the end of the tail. The 3 cm mark on the catheter should be at the anus, and the catheter should not be removed when the knot is tied. Return the rat to its cage after administration.

[0781] The anal plug was left in the rectum for 4 hours to prolong the drug's retention time in the animal. The plug was dislodged within 4 hours of drug administration, and the approximate time of dislodgement was recorded.

[0782] Process blood samples as follows: Coat all centrifuge tubes with EDTA-K2 and store them in a refrigerator (2°C–8°C) or an ice-filled cooler, protected from light before use; transfer the collected blood to centrifuge tubes and manually mix by inverting the tubes at least 5 times, then store in an icebox protected from light. Subsequently, within 2 hours of blood collection, centrifuge the samples at 1800g for 10 minutes at 2°C–8°C. After centrifugation, transfer the collected plasma samples to newly labeled centrifuge tubes, divide them into two equal groups, and store them protected from light at below -70°C.

[0783] Plasma concentrations of mesalazine were analyzed using LC-MS / MS. AUC, Cmax, Cmax, and any other required parameters were calculated using WinNonlin software. Microsoft Office Excel was used for statistical analysis of the data, including mean, standard deviation (SD), and coefficient of variation (CV). Differences in pharmacokinetic parameters between the two groups were compared.

[0784] Detailed pharmacokinetic parameters are shown in Table 18 below. The plasma concentration versus time curve is shown in... Figure 12 middle.

[0785] Table 18

[0786]

[0787] The results showed that the C of mesalazine in group 2 最大 and AUC 最后 The results showed that when mesalazine and sMAP (SEQ ID No:66) were administered in combination in rats, sMAP reduced the absorption and systemic exposure of mesalazine, improved safety, and prolonged the residence time of local application to enhance local efficacy.

[0788] Example 52

[0789] Effects of pre-administration of the compounds of the present invention on the pharmacokinetics of montelukast

[0790] The experiment was conducted in a manner similar to that of Example 51, except that a sodium montelukast suspension was used instead of a mesalazine suppository suspension.

[0791] Montelukast sodium suspension was prepared by weighing an appropriate amount of montelukast sodium and adding it to water to obtain a concentration of 1 mg / mL. A combined formulation was prepared by weighing an appropriate amount of montelukast sodium and synthetic MAP and adding them to water to obtain a suspension containing 1 mg / mL montelukast sodium and 1.5 mg / mL sMAP, and was used immediately after preparation.

[0792] Twelve SD rats (6 males and 6 females) were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. and housed in a barrier facility for 7 days. The temperature was maintained at 20℃-26℃, humidity at 40%-70%, with alternating light and dark cycles for 12 hours, and free access to food and water. As shown in Table 19 below, the rats were randomly divided into two groups of six rats each (3 males and 3 females per group).

[0793] Table 19

[0794]

[0795] Dosage administration, blood sample collection, and pharmacokinetic analysis were performed in the same manner as in Example 51. Table 20 below shows detailed pharmacokinetic parameters, while plasma concentration versus time curves are presented. Figure 13 middle.

[0796] Table 20

[0797]

[0798] The results showed that the C of montelukast in group 2 最大 and AUC 最后 The results showed that when montelukast was administered in combination with sMAP (SEQ ID No:66) in rats, sMAP reduced the absorption and systemic exposure of montelukast, improved safety, and prolonged the residence time of local application to enhance local efficacy.

[0799] Example 53

[0800] Pre-administration of the compounds of this invention to influence the stability and / or pharmacokinetics of different drugs.

[0801] A sMAP solution was prepared by weighing 22.5 mg and adding physiological saline to 15 mL to obtain a concentration of 1.5 mg / mL. Rats were treated by administering the test samples. Test sample 1 was a 5-aminosalicylic acid (mesalazine) suppository, which was melted in a water bath at 40°C. Test sample 2 was montelukast sodium gel, which was prepared as described in Example 52.

[0802] Eighteen SD rats (nine males and nine females) were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. and housed in a barrier facility for 7 days. The temperature was maintained at 20℃-26℃, humidity at 40%-70%, with 12-hour light-dark cycles, and free access to food and water. As shown in Table 21, the rats were randomly divided into three groups of six rats each (three males and three females per group).

[0803] Table 21

[0804] Group deal with dose level Dose Volume Dosage route 1 Mesalazine 1g 200μl Rectal administration 2 Mesalazine + sMAP 1g 200μl Rectal administration 3 Montlust + sMAP 1mg 200μl Rectal administration

[0805] Rats were anesthetized by inhalation of 2% isoflurane. The rats were placed in a supine position, and feces at the end of the rectum were squeezed out. The skin area around the anus was disinfected using 75% alcohol swabs. Group 1 was administered a suspension of 5-aminosalicylic acid (mesalazine) suppositories directly, while groups 2 and 3 were administered montelukast sodium gel and a suspension of 5-aminosalicylic acid (mesalazine) suppositories (respectively) 15 min after treatment with an enema of MAP solution (details of enema dosage and method are in Table 17 below).

[0806] Table 22

[0807] Group deal with Dosage level ( / rat) Dosage-volume ratio (per rat) 1 Mesalazine —— —— 2 Mesalazine + sMAP 1.5mg 1000μl 3 Montlust + sMAP 1.5mg 1000μl

[0808] Use a 2ml syringe to aspirate the air bag cavity to ensure there is no air left in the air bag after filling with water. Insert the catheter (2-WAY 8Ch / Fr 3-5mm) into the rat's anus to a depth of approximately 3cm (2cm from the end of the balloon; cut the tip of the catheter below the tip of the urethral catheter and shorten it as much as possible while maintaining the integrity of the air cavity. Remove the guidewire and seal the urethral catheter with neutral silicone sealant, ensuring a smooth seal to minimize damage to the rectal mucosa). Using a 1mL syringe, draw up a suspension of mesalazine sodium gel (UP-611 gel (5mg / g)) and 5-aminosalicylic acid (mesalazine) suppositories (separately) onto a disposable feeding needle (soft needle) and insert it (approximately 3.5cm) into the anus. Slowly inject 200μL of the syringe contents into each rat.

[0809] While maintaining the perianal skin to secure the catheter and feeding needle, inject 1 mL of water into the air sac cavity using a 2 mL syringe to inflate it, and then quickly release the needle to observe for any gel leakage.

[0810] After injection, secure the catheter by wrapping it around the base of the tail with tape (lift the catheter approximately 3-5 cm from the anus, then cut it after connection and secure it with tape around the base of the tail). The pressure-sensitive tape should be applied based on the non-relaxation standard of the urinary catheter; the catheter should not be too tight and should not cause the animal to chew or feel uncomfortable. The catheter should be secured with tape twice: first, the catheter should be knotted, and second, the remaining catheter should be secured to the end of the tail. The 3 cm mark on the catheter should be at the anus, and the catheter should not be removed when the knot is tied. Return the rat to its cage after administration.

[0811] The anal plug was left in the rectum for 4 hours to prolong the drug's retention time in the animal. The plug was dislodged within 4 hours of drug administration, and the approximate time of dislodgement was recorded.

[0812] Process blood samples as follows: Coat all centrifuge tubes with EDTA-K2 and store them in a refrigerator (2°C–8°C) or an ice-filled cooler, protected from light before use; transfer the collected blood to centrifuge tubes and manually mix by inverting the tubes at least 5 times, then store in an icebox protected from light. Subsequently, within 2 hours of blood collection, centrifuge the samples at 1800 rpm for 10 minutes at 2°C–8°C. After centrifugation, transfer the collected plasma samples to newly labeled centrifuge tubes, divide them into two equal groups, and store them protected from light at below -70°C.

[0813] Pharmacokinetic Analysis: Plasma concentrations of 5-aminosalicylic acid (mesalazine) and montelukast were analyzed using LC-MS / MS and relevant standard operating procedures (SOPs). AUC, C-maximum, T-maximum, and any other required parameters were calculated using WinNonlin software. Microsoft Office Excel was used for statistical analysis of the data, including mean, standard deviation (SD), and coefficient of variation (CV). Differences in pharmacokinetic parameters were compared between the control group receiving the MAP enema and the control group not receiving the MAP enema. Detailed analytical documentation is retained in the study file.

[0814] Example 54

[0815] The compounds of this invention are used as coatings.

[0816] Prepare 0.01-0.5M buffer solutions with different pH values ​​(6.0-9.0), including 0.01M sodium phosphate (pH 6.0), 0.1M sodium bicarbonate (pH 8.5), 0.2M sodium carbonate (pH 9.0), and 0.5M sodium phosphate (pH 7.0).

[0817] Add the buffer solution to the container, and the container surface will be coated (e.g., a cell culture surface). Calculate the volume of the surface to be immersed and the total area already covered by the buffer solution. Cover 90% of the total volume (covered surface) with the buffer solution, while the remaining 10% of the total volume is covered by a solution of one or more compounds of the present invention (such as SEQ ID No: 48, 51, 54, 60, 61, 66, 67, 72, 73, etc.) (at different concentrations, e.g., 1 μg / mL to 100 mg / mL) or in a pre-crosslinked form (see above).

[0818] Thoroughly mix the buffer solution and the compound of the present invention (i.e., the coating solution) in the container and incubate at 4°C–80°C for approximately 10 minutes to 48 hours to coat the surface. Then, pour out or transfer the coating solution and wash the surface with water (using the same total volume as the coating solution).

[0819] The total amount (mg) of added sMAP or pre-crosslinked sMAP can be divided by the total area (cm²) already covered by the solution. 2 To roughly calculate the coating density (mg / cm³) 2 ).

[0820] Example 55

[0821] {[(DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys}2-Lys(SEQ Cell culture plates coated with ID No. 70

[0822] Using a method substantially the same as that described in Example 54 above, a 0.1M sodium bicarbonate (pH 8.5) solution was freshly prepared by dissolving 8.4g of sodium bicarbonate in pure water to a total volume of 1L. The pH of the solution was approximately 8.5.

[0823] Prepare two cornering Costar 24-well cell culture plates. A total solution volume of 400 μL per well is sufficient to cover the bottom surface of the cell culture plate. The total surface area to be covered by the solution is approximately 3 cm². 2 Add 360 μL of 0.1 M sodium bicarbonate (pH 8.5) solution (prepared as above) to each well to be coated.

[0824] 10 mL of 2.25 mg / mL and 1.125 mg / mL peptides were prepared by dissolving 22.5 mg and 11.25 mg of peptides in pure water to a total volume of 10 mL, respectively. {[(DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys ]2-Lys}2-Lys(SEQ ID No: 70) Solution. Add 40 μL of 2.25 mg / mL peptide solution to one cornering costar 24-well cell culture plate pre-filled with 0.1 M sodium bicarbonate (pH 8.5) solution and mix well. Add 40 μL of 1.125 mg / mL peptide solution to the other cornering costar 24-well cell culture plates pre-filled with 0.1 M sodium bicarbonate (pH 8.5) solution and mix well. Then, place both plates at room temperature to allow the solution to coat for 18 hours. Pour off the coating solution and wash the coated wells once with the same volume of pure water as the coating solution.

[0825] The total amount of peptides added (mg) can be divided by the total area (cm²) already covered by the solution. 2 This allows for the approximate calculation of the coating density (mg / cm³) of the coated cell culture plate. 2 Therefore, the coating density is 30 μg / cm³. 2 and 15μg / cm 2 .

[0826] Example 56

[0827] The compound of this invention serves as a carrier for heparin sodium used in infusion management.

[0828] A 1 mg / mL solution of [(DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys)2-Lys]2-Lys (SEQ ID No: 64) was obtained by dissolving it in purified water using the method described in Example 55 of the foregoing embodiments.

[0829] The peptide solution was pumped into the catheter at a flow rate of 0.5 mL / min and left in the catheter for 30 minutes, and then dried at room temperature.

[0830] Heparin sodium powder (185 USP units / mg, Aladdin) was prepared into a 200 μg / mL solution using physiological saline. The heparin sodium solution was pumped into a catheter containing the peptide layer at a flow rate of 0.1 mL / min. The heparin sodium was adsorbed onto the inner wall of the catheter via electrostatic interactions.

[0831] The catheters prepared using the above methods can be used in blood transfusion systems, such as ECMO.

[0832] Example 57

[0833] Preparation of Omeprazole Enteric-Coated Tablets

[0834] Each tablet contains 7.5 mg of sMAP and 20 mg of omeprazole. The formulation is based on 10,000 tablets. Table 21 shows the various steps and the amount of each ingredient in the formulation.

[0835] Table 23

[0836]

[0837]

[0838] Omeprazole granules

[0839] Disodium hydrogen phosphate and Tween 80 were dissolved in purified water by stirring. Omeprazole was added and the mixture was stirred thoroughly. After adding microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose and low-substituted hydroxypropyl cellulose to the mixture, the prepared omeprazole solution was added and stirred thoroughly, granulated, dried and set aside.

[0840] Omeprazole granules with spacer coating

[0841] Hydroxypropyl methylcellulose was slowly added to purified water and stirred until it became clear and transparent. Omeprazole granules were then placed in a coating machine for coating. After coating, the material was dried at 40°C–50°C.

[0842] sMAP particles

[0843] sMAP was dissolved in purified water and then stirred to dissolve it. The prepared amounts of microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose, and low-substituted hydroxypropyl cellulose were mixed, and the prepared sMAP solution was added. The mixture was granulated, dried, and formed into pellets.

[0844] tablet

[0845] After mixing omeprazole-coated granules with sMAP granules, magnesium stearate was added, and the suspension was mixed and then compressed.

[0846] Isolation coat

[0847] Hydroxypropyl methylcellulose was slowly added to purified water, and the mixture was stirred until it became clear and transparent. Omeprazole tablets were then placed in a coating machine for coating and subsequently dried.

[0848] Enteric coating

[0849] Dissolve polyethylene glycol in purified water. Add Eudragit L30D-55, stir the mixture thoroughly, and sieve. Place the core in a coating pan, coat, dry, and sample for testing.

[0850] Example 58

[0851] Preparation of famotidine enteric-coated tablets

[0852] Each tablet contains 7.5 mg of sMAP and 20 mg of famotidine. The formulation is based on 10,000 tablets. Table 22 shows the various steps and the amount of each ingredient in the formulation.

[0853] Table 24

[0854]

[0855]

[0856] Famotidine granules

[0857] Add Tween 80 to purified water, stir and dissolve. Add famodin evenly to the mixture. After adding microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose and low-substituted hydroxypropyl cellulose to the mixture, add the prepared omeprazole solution, stir the mixture evenly, granulate, dry and set aside.

[0858] sMAP particles

[0859] sMAP was dissolved in purified water and then stirred to dissolve. Microcrystalline cellulose, mannitol, lactose, hydroxypropyl methylcellulose and low-substituted hydroxypropyl cellulose were mixed, the prepared sMAP solution was added, granulation was performed, drying was carried out and pelleting was completed.

[0860] tablet

[0861] Mix famotidine granules with sMAP granules. Add magnesium stearate, mix the mixture, and then compress it.

[0862] Isolation coat

[0863] Hydroxypropyl methylcellulose is slowly added to purified water and stirred until it becomes clear and transparent. Famotidine tablets are then placed in a coating machine, coated, and dried.

[0864] Enteric coating

[0865] Polyethylene glycol was dissolved in purified water, Eudragit L30D-55 was added, and the mixture was stirred until homogeneous and then sieved. The core was placed in a coating pan for coating, dried, and samples were taken for testing.

[0866] Example 59

[0867] Preparation of Omeprazole Enteric-Coated Capsules

[0868] Each capsule contains 7.5 mg of sMAP and 20 mg of omeprazole. The formulation is based on a calculation of 10,000 capsules. Table 23 shows the various steps and the amount of each ingredient in the formulation.

[0869] Table 23

[0870]

[0871] Omeprazole granules

[0872] Add disodium hydrogen phosphate and Tween 80 to purified water, stir and dissolve. Add omeprazole and stir the mixture to disperse it evenly. Add microcrystalline cellulose, dextrin, mannitol, hydroxypropyl methylcellulose and low-substituted hydroxypropyl cellulose to the mixture, and then add to the solution for granulation, drying and pelleting. Set aside.

[0873] Omeprazole packing spacer

[0874] Hydroxypropyl methylcellulose was slowly added to purified water and stirred until it became clear and transparent. Omeprazole granules were then placed in a coating machine for coating. After coating, the material was dried at 40°C–50°C.

[0875] sMAP particles

[0876] sMAP was dissolved in purified water and then stirred to dissolve. Microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose and low-substituted hydroxypropyl cellulose were mixed, added to the solution, and then granulated, dried and made into pellets.

[0877] Filled capsules

[0878] Mix omeprazole-coated granules and sMAP granules. Add magnesium stearate, and then fill enteric-coated capsules with the mixture.

[0879] Example 60

[0880] Preparation of famotidine enteric-coated capsules

[0881] Each capsule contains 7.5 mg of sMAP and 20 mg of famotidine. The formulation is based on 10,000 pills. Table 24 below shows the various steps and the amount of each ingredient added to the formulation.

[0882] Table 24

[0883]

[0884]

[0885] Famotidine granules

[0886] Add Tween 80 to purified water and stir the solution to dissolve. Add Modine evenly to the mixture. Mix microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose, and low-substituted hydroxypropyl cellulose, then add to the solution, followed by granulation, drying, and pelletizing. Set aside.

[0887] Famotidine granules with separation layer coating

[0888] Hydroxypropyl methylcellulose was slowly added to purified water and stirred until the mixture became clear and transparent. Famotidine granules were then placed in a coating machine for coating. After coating, the material was dried at 40°C–50°C.

[0889] sMAP particles

[0890] sMAP was dissolved in purified water, and the mixture was then stirred to dissolve. Microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose, and low-substituted hydroxypropyl cellulose were mixed and then added to the solution, followed by granulation, drying, and pelleting.

[0891] Filled capsules

[0892] Mix famotidine granules and sMAP granules. Add magnesium stearate, and then use the mixture to fill enteric-coated capsules.

[0893] Example 61

[0894] Preparation of Omeprazole / sMAP Capsules for Proctitis

[0895] Table 25 below shows the various steps and the amount of each ingredient added to the preparation.

[0896] Table 25

[0897]

[0898] Omeprazole granules

[0899] Add disodium hydrogen phosphate and Tween 80 to purified water. Stir the mixture and dissolve. Stir omeprazole into the mixture and disperse evenly. Mix microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose, and low-substituted hydroxypropyl cellulose, then add to the solution, followed by granulation, drying, and pelleting. Set aside.

[0900] Omeprazole granules with spacer coating

[0901] Hydroxypropyl methylcellulose was slowly added to purified water, and the mixture was stirred until it became clear and transparent. Omeprazole granules were then placed in a coating machine for coating. After coating, the material was dried at 40°C–50°C.

[0902] sMAP particles

[0903] sMAP was dissolved in purified water and then stirred to dissolve. Microcrystalline cellulose, mannitol, hydroxypropyl methylcellulose and low-substituted hydroxypropyl cellulose were mixed and then added to the solution, followed by granulation, drying and pelleting.

[0904] Filled capsules

[0905] After mixing omeprazole-coated granules and sMAP granules, magnesium stearate was added to the mixture, and then the mixture was used to fill hydroxypropyl methylcellulose empty capsules.

Claims

1. A compound of formula I, in: A and B independently represent Z or A. 1 -Q 1 -B 1 ; Q represents a structural fragment of formula II. in: The wavy line represents the attachment point of Q to A and / or B; and m represents the integer 4; A 1 and B 1 Independently represent Z or A 2 -Q 2 -B 2 ; A 2 and B 2 Independently represent Z or ZQ 3 -Z; Q 1 Q 2 and Q 3 Independently representing the structural fragment of Formula III, in: The wavy lines adjacent to the NH group represent Q. 1 Q 2 and Q 3 With A 1 and / or B 1 A 2 and / or B 2 The attachment point of Z; The wavy lines adjacent to C=O represent Q. 1 Q 2 and Q 3 With Q, Q 1 and Q 2 The attachment point; and m is as defined above; In each case where it is used, Z represents a peptide component having the following amino acid sequence: [W-Lys-X 1 -Ser-U-X 2 -Y] n -W-Lys-X 1 -Ser-U-X 2 -Y---(SEQ ID No: 3) in: The dashed lines represent the attachment points of Z to the rest of the molecule; n represents 0; and In each case where they are used: W is selected from the groups HCA, HCA-Ala-, Ala, DOPA, Lys-Ala-, and DOPA-Ala-; X 1 It represents Pro or Hyp; U stands for Tyr or DOPA; X 2 It represents Hyp; and Y represents a 4-amino acid sequence, wherein the amino acids are selected from one or more of the following groups: Lys, Ala, Hyp, Thr, DOPA, and Tyr. And the regioisomers, stereoisomers, and pharmaceutically or cosmetically acceptable salts of the compound.

2. The compound according to claim 1, wherein A and B both represent Z or both represent A. 1 -Q 1 -B 1 .

3. The compound according to claim 1, wherein A 1 and B 1 Both represent Z or both represent A 2 -Q 2 -B 2 .

4. The compound according to claim 1, wherein A 2 and B 2 All represent Z or all represent ZQ 3 -Z.

5. The compound according to claim 1, wherein X 1 It stands for Pro.

6. The compound according to claim 5, wherein Y represents an amino acid sequence selected from the group consisting of: -Hyp-Y 1 -Y 2 -Lys- and -Thr-Y 1 -Y 2 -Lys-, where Y 1 and Y 2 Each of them is independently selected from the following groups: Ala, Hyp, Thr, DOPA, and Tyr.

7. The compound according to claim 5, wherein the amino acid sequence defined by Y is selected from the group consisting of -Hyp-Thr-Tyr-Lys-, -Hyp-Thr-DOPA-Lys-, -Hyp-Thr-Ala-Lys-, -Thr-Tyr-Hyp-Lys-, -Thr-DOPA-Hyp-Lys- and -Thr-Ala-Hyp-Lys-.

8. The compound according to claim 1, wherein U represents Tyr.

9. The compound according to claim 1, wherein W represents Ala or Lys-Ala-.

10. The compound according to claim 1, wherein Z is selected from the group consisting of: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 2); Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys--- (SEQ ID No: 1); Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 4); and Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys--- (SEQ ID No: 5).

11. The compound according to claim 1, wherein W represents HCA, HCA-Ala-, DOPA, or DOPA-Ala-.

12. The compound according to claim 11, wherein Z is selected from the group consisting of: HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 12); HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 13); HCA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys--- (SEQ ID No: 14); HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 15); DOPA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 16); DOPA-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 17); DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 18); and DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 19).

13. The compound according to claim 1, wherein U represents DOPA.

14. The compound according to claim 13, wherein Z is selected from the group consisting of: Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 20); and Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys--- (SEQ ID No: 21).

15. The compound according to claim 1, wherein Z is selected from the group consisting of: HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 26); HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys--- (SEQ ID No: 27); HCA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 28); HCA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 29); DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 30); DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 31); DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 32); DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 33;and DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys--- (SEQ ID No: 34).

16. The compound according to claim 1, wherein both A and B represent Z, and one or both Z groups represent: Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 4), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 5), HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 12), HCA-Lys-Pro-Ser-Tyr-Hyp-Thr-Ala-Hyp-Lys--- (SEQ ID No: 15), DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys—(SEQ ID No: 18), Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 20), HCA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 26), DOPA-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-Ala-Lys---(SEQ ID No: 31), DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 32), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 2), or Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 1); and Q represents the Lys fragment.

17. The compound according to claim 1, wherein both A and B represent A. 1 -Q 1 -B 1 A 1 and B 1 All represent Z, one or two Z groups represent: Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 4), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 5), HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 12), HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 13), DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 18), Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 20), Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 21), DOPA-Lys-Pro-Ser-DOPA-Hyp-Thr-Ala-Hyp-Lys---(SEQ ID No: 30), DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 32), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 2), or Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 1); and Q 1 Represents the Lys fragment.

18. The compound according to claim 1, wherein both A and B represent A. 1 -Q 1 -B 1 A 1 and B 1 All represent A 2 -Q 2 -B 2 A 2 and B 2 All represent Z, one or two Z groups represent: Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 4), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 5), HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 12), DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 18), Ala-Lys-Pro-Ser-DOPA-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 21), DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 32), DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys---(SEQ ID No: 19), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 2), or Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys---(SEQ ID No: 1); and Q 1 and Q 2 All of these represent Lys segments.

19. The compound according to claim 1, wherein both A and B represent Z and both Z groups represent: Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-Tyr-Lys--- (SEQ ID No: 2), Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Thr-DOPA-Lys--- (SEQ ID No: 1), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys---(SEQ ID No: 4), Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-DOPA-Hyp-Lys---(SEQ ID No: 5), HCA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 12), DOPA-Ala-Lys-Pro-Ser-Tyr-Hyp-Thr-Tyr-Hyp-Lys--- (SEQ ID No: 18), Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys--- (SEQ ID No: 20), or DOPA-Ala-Lys-Pro-Ser-DOPA-Hyp-Thr-DOPA-Hyp-Lys--- (SEQ ID No: 32).

20. A pharmaceutical formulation comprising the compound according to any one of claims 1 to 19.

21. The pharmaceutical formulation of claim 20, further comprising a pharmaceutically or cosmetically acceptable adjuvant, diluent, or carrier.

22. The pharmaceutical formulation of claim 20, which is suitable for, adapted to and / or packaged for, and presented for external application, wherein the pharmaceutically or cosmetically acceptable adjuvant, diluent or carrier is a topical adjuvant, diluent or carrier.

23. The pharmaceutical preparation according to claim 20, wherein it is in the form of a gel, spray, cream, ointment or powder.

24. The pharmaceutical preparation according to claim 20, further comprising one or another pharmaceutically active ingredient.

25. A kit comprising the following components: (A) the compound according to any one of claims 1 to 19 or the pharmaceutical preparation according to any one of claims 20 to 23; and (B) A pharmaceutical formulation comprising one or more active pharmaceutical ingredients mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier. The components (A) and (B) are each provided in a form suitable for combined application with each other.

26. The pharmaceutical formulation of claim 24 or the kit of claim 25, wherein the active pharmaceutical ingredient is an anti-inflammatory agent, a pro-inflammatory agent, an antibiotic, an antibacterial agent and / or an antiprobiotic agent, an antiviral agent, an anesthetic agent and / or a wound healing agent.

27. The pharmaceutical formulation or kit according to claim 26, wherein the active pharmaceutical ingredient is an anti-inflammatory agent.

28. The use of the compound according to any one of claims 1 to 19, the formulation according to any one of claims 20 to 24, 26 or 27, or the kit according to any one of claims 25 to 27 in the manufacture of a medicament for treating inflammation, inflammatory disorders, and / or disorders characterized by inflammation.

29. The use according to claim 28, wherein the inflammatory barrier is a wound or burn or causes a wound.

30. The use according to claim 29, wherein the obstruction causing the wound is hemorrhoids or ulcerative colitis.

31. The pharmaceutical preparation according to claim 20, wherein the one or more compounds or salts thereof are applied topically as an external preparation.

32. The pharmaceutical preparation according to claim 31, wherein the relevant condition is treated by direct topical application to the skin.

33. The pharmaceutical preparation according to claim 31, wherein the relevant condition is treated by direct topical application to a mucosal surface.

34. The pharmaceutical formulation of claim 20, wherein the one or more compounds are administered orally, intravenously, through the skin or subcutaneously, via the nose, intramuscularly, intraperitoneally, via the lungs, or via the anus or rectum.

35. The pharmaceutical formulation according to claim 24, wherein the compound according to any one of claims 1 to 19 functions as an excipient, a medical device, or a component of a pharmaceutical-medical device combination.

36. The pharmaceutical formulation of claim 24, wherein the compound is any one of claims 1 to 7 or 11 to 19, wherein W is according to claim 11 and / or U is according to claim 13, and the compound is crosslinked before or after administration to a subject.

37. The kit according to claim 25, wherein the one or more compounds or salts thereof are applied topically as an external formulation.

38. The kit of claim 37, wherein the relevant condition is treated by direct topical application to the skin.

39. The kit of claim 37, wherein the related condition is treated by direct topical application to the mucosal surface.

40. The kit of claim 25, wherein the one or more compounds are administered orally, intravenously, through the skin or subcutaneously, via the nose, intramuscularly, intraperitoneally, via the lungs, or via the anus or rectum.

41. The kit according to claim 25, wherein the compound according to any one of claims 1 to 19 acts as an excipient, a medical device, or a component of a pharmaceutical-medical device combination.

42. The kit of claim 25, wherein the compound is any one of claims 1 to 7 or 11 to 19, wherein W is according to claim 11 and / or U is according to claim 13, and the compound is cross-linked before or after administration to the subject.

43. The use according to claim 28, wherein the one or more compounds or salts thereof are applied topically as an external preparation.

44. The use according to claim 43, wherein the relevant condition is treated by direct topical application to the skin.

45. The use according to claim 43, wherein the related condition is treated by direct topical application to the mucosal surface.

46. ​​The use according to claim 28, wherein the one or more compounds are administered orally, intravenously, through the skin or subcutaneously, via the nose, intramuscularly, intraperitoneally, via the lungs, or via the anus or rectum.

47. The use according to claim 28, wherein the compound according to any one of claims 1 to 19 serves as an excipient, a medical device, or a component of a pharmaceutical-medical device combination.

48. The use according to claim 28, wherein the compound is according to any one of claims 1 to 7 or 11 to 19, wherein W is according to claim 11 and / or U is according to claim 13, and the compound is crosslinked before or after administration to the subject.

49. Use of the compound according to any one of claims 1 to 19 in the preparation of products used as adhesives or film-forming materials.

50. The use according to claim 49, wherein the use is as a wound surface repair product, wound surface protection product, medical bioadhesive product, medical coating product, industrial coating product, biochemical reagent, medical product, sterilization product, or culture container for cell culture.

51. The use according to claim 49, wherein a film is formed on the surface of a skin or mucous membrane wound to aid in recovery.

52. The use according to claim 49, wherein the use is for closing surgical incisions, adhering to fractured bones, adhering to mucous membranes, or covering human implants.

53. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 19 and a pharmaceutically or cosmetically acceptable adjuvant, diluent, or carrier, which serves as an adhesive or film-forming material.

54. The pharmaceutical formulation of claim 53, which is suitable for, adapted to and / or packaged for, and presented for external application, wherein the pharmaceutically or cosmetically acceptable adjuvant, diluent or carrier is a topical adjuvant, diluent or carrier.

55. The pharmaceutical preparation according to claim 53, wherein it is in the form of a gel, spray, cream, ointment or powder.

Images