Heme derivatives and their preparation and use

Abstract

A heme derivative and its preparation method and use. Specifically, a heme derivative as shown in formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof is provided. The above heme derivative has excellent peroxidase activity, and therefore can be used to scavenge free radicals and to treat or prevent diseases associated with free radicals. Z1-Z2(I)

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry and pharmacotherapy, and specifically relates to heme derivatives and preparation methods and uses thereof. Background Art

[0002] Peroxidase is widely present in animals, plants and aerobic microorganisms. It can catalyze hydrogen peroxide, oxidize phenolic and amine compounds and hydrocarbon oxidation products, and has the dual role of eliminating the toxicity of hydrogen peroxide and phenols, amines, aldehydes and benzene.

[0003] Peroxidase is of great significance for the prevention and treatment of various diseases caused by free radical damage. Although natural peroxidase is an effective antioxidant drug, it has the disadvantages of limited sources, difficult purification, poor stability, large molecular weight, and short half-life in vivo. Therefore, people try to use enzyme mimics to prepare peroxidase mimics with medicinal value.

[0004] Octapeptide (MP-8), nonapeptide (MP-9), and undecapeptide (MP-11) hydrolyzed from cytochrome C (Cyt C) all contain covalently bound heme and a histidine residue in the peptide segment. They are found to be a good class of peroxidase mimics with good antioxidant activity. People have conducted a large number of studies on them as peroxidase mimics, showing that they have significant preventive and therapeutic effects on a variety of diseases related to free radical damage. Since the heme molecules on MP-8, MP-9, and MP-11 are connected to the short peptide through a thioether bond, they are not easy to synthesize chemically in vitro. They are generally obtained by in vitro enzymatic hydrolysis of cytochrome C and further purification. There are many purification steps, low purity, and they are easily degraded in vitro, which limits their medicinal value.

[0005] In summary, there is an urgent need in the art to develop a new class of peroxidase mimetics that are easy to synthesize artificially and stable in vitro. Summary of the invention

[0006] The purpose of the present invention is to provide a new type of peroxidase mimics that are easy to synthesize artificially and stable in vitro.

[0007] In the first aspect of the present invention, there is provided a heme derivative as shown in Formula I or a pharmaceutically acceptable salt, enantiomer, diastereomer or racemate thereof,

[0008] Z1-Z2 (I)

[0009] In the formula, Z1 is a substituted or unsubstituted porphyrin ring, wherein the porphyrin ring forms a planar coordination structure with the metal M; Z2 is a first coordination structure, wherein the first coordination structure further forms a stereo coordination structure with the metal M; and “-” is a chemical bond.

[0010] In another preferred embodiment, the first coordination structure has an imidazole group, and the imidazole group and the metal M form a stereo coordination structure.

[0011] In another preferred embodiment, the imidazole group is 4-imidazolyl, more preferably

[0012] In another preferred embodiment, the heme derivative is as shown in Formula Ia

[0013]

[0014] In the formula,

[0015] M is a coordination metal, and M represents a metal element selected from the eighth group, the first subgroup, the second subgroup and the seventh subgroup of the periodic table (preferably, M represents a metal element selected from the following group: iron, manganese, cobalt, nickel and zinc; more preferably, M represents a metal element selected from the following group: iron, manganese, zinc); more preferably, M is a divalent or trivalent metal;

[0016] X is selected from the group consisting of -C(R')2-, -N(R')-, -O-, -S-;

[0017] Y is each independently selected from the group consisting of -C(R')2-, -N(R')-, -C(O)-, -O-, -S-;

[0018] n = 0, 1 or 2;

[0019] The additional condition is: -X-(Y) n - is of chemically stable structure;

[0020] R1 and R2 are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3 to 7 membered heterocyclyl, substituted or unsubstituted C6-C10 aryl (preferably phenyl), substituted or unsubstituted 5 to 10 membered heteroaryl; or, R1 and R2 and the carbon to which they are attached together form a substituted or unsubstituted C3-C7 cycloalkyl (preferably C3-C5 cycloalkyl) or a 3 to 7 membered heterocyclyl;

[0021] A ais an amino acid residue or a divalent linking group having 1 to 5 chain atoms (preferably, a divalent linking group having 3 or 4 chain atoms);

[0022] R t is selected from the group consisting of H, hydroxy, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3-7 membered heterocyclyl;

[0023] R 4a Selected from the following group: H, hydroxyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2;

[0024] R 5a , R 5a '、R 6a and R 6a 'Each is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted -C(O)-C1-C6 alkyl;

[0025] R' is independently selected from: H, substituted or unsubstituted C1-C6 alkyl;

[0026] Unless otherwise specified, the substitution refers to the replacement of one or more (preferably 1 to 3) hydrogen atoms in the group by a substituent selected from the following group: halogen (preferably, the halogen is F, C1 or Br), hydroxyl, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2 (preferably dimethylamino), C1-C6 alkoxy (preferably methoxy or ethoxy), -S-C1-C6 alkyl, -O-C1-C6 haloalkyl, -S-C1-C6 haloalkyl.

[0027] In another preferred embodiment, the divalent linking group having 1 to 5 chain atoms is a C1-C5 alkylene group.

[0028] In another preferred embodiment, A a -R t Selected from: -C1-C5 alkylamino, -C1-C5 alkyloxy, amino acid residue -NH2, amino acid residue -OH, amino acid residue -O-C1-C6 alkyl, amino acid residue -NH-C1-C6 alkyl.

[0029] In another preferred embodiment, -X-(Y) n - is selected from the group consisting of -N(R')-, -N(R')-C(R')2-, and -N(R')-C(R')2-C(R')2-.

[0030] In another preferred embodiment, the amino acid residue is derived from a natural amino acid or a non-natural amino acid.

[0031] In another preferred embodiment, the amino acid residue is derived from D-type and / or L-type amino acids.

[0032] In another preferred embodiment, the amino acid residue is a residue derived from the following group of amino acids: Ala (A), Arg (R), Asn (N), Asp (D), Cys (C), Gln (Q), Glu (E), Gly (G), His (H), Ile (I), Leu (L), Lys (K), Met (M), Phe (F), Pro (P), Ser (S), Thr (T), Trp (W), Tyr (Y), Val (V).

[0033] In another preferred embodiment, the amino acid residue is a residue derived from an amino acid selected from the following group: L-glycine (L-Gly), L-alanine (L-Ala), β-alanine (β-Ala), L-glutamic acid (L-Glu), L-aspartic acid (L-Asp), L-histidine (L-His), L-arginine (L-Arg), L-lysine (L-Lys), L-valine (L-Val), L-serine (L-Ser), L-threonine (L-Thr).

[0034] In another preferred embodiment, the amino acid residue refers to a group formed by removing one H from the N-terminal -NH2 of the amino acid and removing -OH from the C-terminal -COOH.

[0035] In another preferred embodiment, -X-(Y) n -C(R1)R2-CO- constitutes an amino acid residue structure.

[0036] In another preferred embodiment, the amino acid residue is derived from a natural amino acid or a non-natural amino acid.

[0037] In another preferred embodiment, the amino acid residue is derived from D-type and / or L-type amino acids.

[0038] In another preferred embodiment, in Formula Ia, R 5a and R 5a ' are the same group.

[0039] In another preferred embodiment, in Formula Ia, R 6a and R6a ' are the same group.

[0040] In another preferred embodiment, in Formula Ia,

[0041] R 5a and R 5a ' are both H, and R 6a and R 6a ' are each independently selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted -C(O)-C1-C6 alkyl (preferably, R 6a and R 6a ' is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl); or,

[0042] R 6a and R 6a ' are both H, and R 5a and R 5a ' are each independently selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted -C(O)-C1-C6 alkyl (preferably, R 6a and R 6a ' is selected from the group consisting of substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl).

[0043] In another preferred embodiment, in Formula Ia, R1 and R2 are each independently selected from the following group: H, methyl, ethyl, propyl, butyl, and phenyl.

[0044] In another preferred embodiment, in Formula Ia, at least one of R1 and R2 is hydrogen or methyl.

[0045] In another preferred embodiment, in Formula Ia, R1 and R2 are both hydrogen.

[0046] In another preferred embodiment, in Formula Ia, It is S configuration, R configuration, or a combination thereof; preferably, it is S configuration; and wherein R2 is hydrogen, and R1 is selected from the following group: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3 to 7 membered heterocyclyl (preferably, R1 is selected from the following group: methyl, ethyl, propyl, butyl, phenyl).

[0047] In another preferred embodiment, in Formula Ia, It is S configuration, R configuration, or a combination thereof; preferably, it is S configuration, R configuration, or a combination thereof; preferably, it is S configuration.

[0048] In another preferred embodiment, in Formula Ia, X is selected from the following group: -NH-, -O-.

[0049] In another preferred embodiment, the heme derivative as shown in Formula Ia can have a structure as shown in Formula Ia-i

[0050]

[0051] in,

[0052] Dashed lines refer to: none, coordination bond, covalent bond, or hydrogen bond;

[0053] X, Y, R1, R2, R t ,M,n,R 4a , R 5a , R 5a '、R 6a , R 6a ' and A a The definition of is the same as that in Formula Ia.

[0054] In another preferred embodiment, A a , R', X, Y, R1, R2, R t , M and n are corresponding groups in Table A(a) or Table C or in the compounds of formula I prepared in the Examples.

[0055] In another preferred embodiment, R 4a , R 5a , R 5a '、R 6a and R 6a ' is the group corresponding to the compound shown in Table A(a).

[0056] In another preferred embodiment, the heme derivative as shown in Formula Ia is selected from Table A(a); Table A(a)

[0057]

[0058]

[0059]

[0060] In another preferred embodiment, the heme derivative is as shown in Formula Ib-1 or Formula Ib-2;

[0061]

[0062] In various

[0063] M is a coordination metal, and M represents a metal element selected from the eighth group, the first subgroup, the second subgroup and the seventh subgroup of the periodic table (preferably, M represents a metal element selected from the following group: iron, manganese, cobalt, nickel and zinc; more preferably, M represents a metal element selected from the following group: iron, manganese, zinc); more preferably, M is a divalent or trivalent metal;

[0064] X is selected from the group consisting of -C(R')2-, -N(R')-, -O-, -S-;

[0065] Y is each independently selected from the group consisting of -C(R')2-, -N(R')-, -C(O)-, -O-, -S-;

[0066] n = 0, 1 or 2;

[0067] The additional condition is: -X-(Y) n - is of chemically stable structure;

[0068] R1 and R2 are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3 to 7 membered heterocyclyl, substituted or unsubstituted C6-C10 aryl (preferably phenyl), substituted or unsubstituted 5 to 10 membered heteroaryl; or, R1 and R2 and the carbon to which they are attached together form a substituted or unsubstituted C3-C7 cycloalkyl (preferably C3-C5 cycloalkyl) or a 3 to 7 membered heterocyclyl;

[0069] R 3b For-(A m ) o -R t ;in,

[0070] A m is an amino acid residue or a divalent linking group having 1 to 5 chain atoms (preferably 3 or 4 chain atoms);

[0071] R t is selected from the group consisting of H, hydroxy, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3-7 membered heterocyclyl;

[0072] o=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0073] R 4bSelected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2;

[0074] R 5b , R 5b '、R 6b and R 6b 'Each is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -C(O)-C1-C6 alkyl;

[0075] R 7b and R 7b 'Each is independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted -C(O)-C1-C6 alkyl;

[0076] R' is independently selected from: H, substituted or unsubstituted C1-C6 alkyl;

[0077] Unless otherwise specified, the substitution refers to the replacement of one or more (preferably 1 to 3) hydrogen atoms in the group by a substituent selected from the following group: halogen (preferably, the halogen is F, Cl or Br), hydroxyl, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2 (preferably dimethylamino), C1-C6 alkoxy (preferably methoxy or ethoxy), -S-C1-C6 alkyl, -O-C1-C6 haloalkyl, -S-C1-C6 haloalkyl.

[0078] In another preferred embodiment, the divalent linking group having 1 to 5 chain atoms is a C1-C5 alkylene group.

[0079] In another preferred embodiment, (A m ) o -R t Selected from: C1-C18 alkylamino, C1-C18 alkyloxy, (amino acid residue) o -NH2, (amino acid residue) o -OH, (amino acid residue) o -O-C1-C6 alkyl, (amino acid residue) o -NH-C1-C6 alkyl.

[0080] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, R 5b and R 5b ' are the same group.

[0081] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, R 6b and R 6b ' are the same group.

[0082] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, R 5b and R 5b 'All H, or R 6b and R 6b 'All are H.

[0083] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, R 7b and R 7b ' are the same group.

[0084] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, R 7b and R 7b 'All are H.

[0085] In another preferred embodiment, the definition of the amino acid residue is the same as that of the amino acid residue in formula Ia.

[0086] In another preferred embodiment, -X-(Y) n -C(R1)R2-CO- constitutes an amino acid residue structure.

[0087] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, R1 and R2 are defined as in Formula Ia.

[0088] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, S configuration, R configuration, or a combination thereof; preferably, S configuration; and wherein R1 and R2 are defined as in Formula Ia.

[0089] In another preferred embodiment, in Formula Ib-1 and / or Formula Ib-2, It is S configuration, R configuration, or a combination thereof; preferably, it is S configuration, R configuration, or a combination thereof; preferably, it is S configuration.

[0090] In another preferred embodiment, the heme derivatives shown in Formula Ib-1 or Formula Ib-2 can have structures shown in Formula Ib-1i and Ib-2i, respectively.

[0091]

[0092] in,

[0093] Dashed lines refer to: none, coordination bond, covalent bond, or hydrogen bond;

[0094] X, Y, R1, R2, M, n, R 3b , R4b , R 5b , R 5b '、R 6b , R 6b '、R 7b and R 7b The definition of ' is the same as that in Formula Ib-1 and / or Ib-2.

[0095] In another preferred embodiment, R', o, A m ,X,Y,R1,R2,R t , M and n are corresponding groups in the compounds of formula I in Table A(b) or Table C or prepared in the Examples.

[0096] In another preferred embodiment, R 3b , R 4b , R 5b , R 5b '、R 6b , R 6b '、R 7b and R 7b ' is the group corresponding to the compound shown in Table A(b).

[0097] In another preferred embodiment, the heme derivative represented by Formula Ib-1 or Formula Ib-2 is selected from Table A(b);

[0098] Table A(b)

[0099]

[0100]

[0101] In another preferred embodiment, the heme derivative is as shown in Formula Ic;

[0102]

[0103] In the formula,

[0104] M is a coordination metal, and M represents a metal element selected from the eighth group, the first subgroup, the second subgroup and the seventh subgroup of the periodic table (preferably, M represents a metal element selected from the following group: iron, manganese, cobalt, nickel and zinc; more preferably, M represents a metal element selected from the following group: iron, manganese, zinc); more preferably, M is a divalent or trivalent metal;

[0105] X is selected from the group consisting of -C(R')2-, -N(R')-, -O-, -S-;

[0106] Y is each independently selected from the group consisting of -C(R')2-, -N(R')-, -C(O)-, -O-, -S-;

[0107] n = 0, 1 or 2;

[0108] The additional condition is: -X-(Y) n - is of chemically stable structure;

[0109] R1 and R2 are each independently selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3 to 7 membered heterocyclyl, substituted or unsubstituted C6-C10 aryl (preferably phenyl), substituted or unsubstituted 5 to 10 membered heteroaryl; or, R1 and R2 and the carbon to which they are attached together form a substituted or unsubstituted C3-C7 cycloalkyl (preferably C3-C5 cycloalkyl) or a 3 to 7 membered heterocyclyl;

[0110] R 3c -(Am) o -R t ;in,

[0111] A m is an amino acid residue or a divalent linking group having 1 to 5 chain atoms (preferably 3 or 4 chain atoms);

[0112] R t is selected from the group consisting of H, hydroxy, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C5 cycloalkyl, substituted or unsubstituted 3-7 membered heterocyclyl;

[0113] o=0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0114] R 4c Selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted amino, substituted or unsubstituted NH(C1-C6 alkyl), substituted or unsubstituted N(C1-C6 alkyl)2;

[0115] R 5c and R 5c 'Each independently selected from the following group: H, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkyl-C(O) (preferably, each independently selected from the following group: H, substituted or unsubstituted C2-C6 alkenyl);

[0116] R' is independently selected from: H, substituted or unsubstituted C1-C6 alkyl;

[0117] Unless otherwise specified, the substitution refers to the replacement of one or more (preferably 1 to 3) hydrogen atoms in the group by a substituent selected from the following group: halogen (preferably, the halogen is F, Cl or Br), hydroxyl, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2 (preferably dimethylamino), C1-C6 alkoxy (preferably methoxy or ethoxy), -S-C1-C6 alkyl, -O-C1-C6 haloalkyl, -S-C1-C6 haloalkyl.

[0118] In another preferred embodiment, R 3c Same definition as R 3b .

[0119] In another preferred embodiment, in Formula Ic, R 5c and R 5c ' are the same group.

[0120] In another preferred embodiment, in Formula Ic, R 5c and R 5c ' are H or substituted or unsubstituted C2-C6 alkenyl.

[0121] In another preferred embodiment, the amino acid residue is defined as the amino acid residue in Formula Ia.

[0122] In another preferred embodiment, -X-(Y) n -C(R1)R2-CO- constitutes an amino acid residue structure.

[0123] In another preferred embodiment, in Formula Ic, R1 and R2 are defined as in Formula Ia.

[0124] In another preferred embodiment, in Formula Ic, It is S configuration, R configuration, or a combination thereof; preferably, it is S configuration; and wherein R1 and R2 are defined as in Formula Ia.

[0125] In another preferred embodiment, in Formula Ic, It is S configuration, R configuration, or a combination thereof; preferably, it is S configuration, R configuration, or a combination thereof; preferably, it is S configuration.

[0126] In another preferred embodiment, the heme derivative as shown in formula Ic can have a structure as shown in formula Ic-i

[0127]

[0128] in,

[0129] Dashed lines refer to: none, coordination bond, covalent bond, or hydrogen bond;

[0130] X, Y, R1, R2, M, n, R 3c , R 4c , R 5c and R 5c The definition of ' is the same as that in Formula Ib-1 and / or Ib-2.

[0131] In another preferred embodiment, R', R t , o, Am, X, Y, R1, R2, M and n are corresponding groups in the compounds of formula I in Table A(c) or Table C or prepared in the Examples.

[0132] In another preferred embodiment, R 3b , R 4b , R 5b , R 5b '、R 6b , R 6b '、R 7b and R 7b ' is the group corresponding to the compound shown in Table A(c).

[0133] In another preferred embodiment, the heme derivative as shown in Formula Ic is selected from Table A(c);

[0134] Table A(c)

[0135]

[0136]

[0137]

[0138]

[0139] In a second aspect of the present invention, there is provided a compound of formula IIa,

[0140]

[0141] In the formula,

[0142] A a ' is an amino acid residue, or a divalent linking group having 1 to 5 chain atoms (preferably 3 or 4 chain atoms);

[0143] p = 1, 2, 3, 4, 5, 6, 7, 8 or 9;

[0144] And-(A a ') p - at least one end is -NR'-, -O- or -S-; and -(Aa ') p -Through the NR'-, -O- or -S- a connect;

[0145] R',M,X,Y,n,R1,R2,R t , A a , R 4a , R 5a , R 5a '、R 6a and R 6a ' is defined as in Formula Ia.

[0146] In another preferred embodiment, the compound of formula IIa is a prodrug of a heme derivative as shown in formula Ia.

[0147] In another preferred embodiment, A a ' is an amino acid residue.

[0148] In another preferred embodiment, the amino acid residue is defined as described above.

[0149] In another preferred embodiment, -(A a ')p- at least one end is -NH-; and -(A a ') p -Through the -NH- a connect.

[0150] In another preferred embodiment, A a , A a ', R', X, Y, R1, R2, R t , M and n are corresponding groups in the compounds of formula I in Table B or Table C or prepared in the Examples.

[0151] In another preferred embodiment, p, R 4a , R 5a , R 5a '、R 6a and R 6a ' is the group corresponding to the compound shown in Table B.

[0152] In another preferred embodiment, the compound of formula IIa is selected from the compounds shown in Table B

[0153] Table B

[0154]

[0155]

[0156]

[0157] In a third aspect of the present invention, a pharmaceutical composition is provided, comprising:

[0158] (i) the heme derivative as described in the first aspect and / or the compound of formula IIa as described in the second aspect;

[0159] and (ii) a pharmaceutically acceptable carrier.

[0160] In another preferred embodiment, the heme derivative is as shown in Formula Ia, Formula Ib-1 or Formula Ib-2, or Formula Ic.

[0161] In the fourth aspect of the present invention, there is provided a use of the heme derivative as described in the first aspect and / or the compound of formula IIa as described in the second aspect for preparing a drug or composition for treating or preventing diseases associated with free radicals.

[0162] In another preferred embodiment, the free radical-related diseases include: diseases caused by free radical damage, diseases caused by free radical accumulation and / or diseases caused by peroxide accumulation.

[0163] In another preferred embodiment, the disease associated with free radicals is selected from the following group:

[0164] Complications of diabetes, Oppenheimer's disease, hypertension, Keshan disease, heart failure, atherosclerosis, myocardial infarction, coronary heart disease, acute lung injury, pneumonia, chronic obstructive pulmonary disease, asthma, pulmonary fibrosis, ischemia-reperfusion injury, brain trauma, stroke, Alzheimer's disease, allergic reaction, Hashimoto's disease, lupus erythematosus, dermatomyositis, autoimmune vasculitis, amyotrophic lateral sclerosis, arthritis, cataract, glaucoma, retrolental fibrosis, retinal oxidative damage, optic neuritis, radiation sickness, dermatitis herpetiformis, immersion dermatitis, solar dermatitis, glomerulonephritis, hepatitis, cirrhosis, obesity, diabetes and its complications, tumors, or a combination thereof.

[0165] In another preferred embodiment, the disease associated with free radicals is selected from the following group: ischemia-reperfusion injury, diabetic complications, Oppenheimer's disease, stroke, cataract, or a combination thereof.

[0166] In a fifth aspect of the present invention, there is provided a method for scavenging free radicals, comprising the steps of:

[0167] The subject is incubated with the heme derivative as described in the first aspect and / or the compound of formula IIa as described in the second aspect, thereby clearing free radicals in the subject.

[0168] In another preferred embodiment, the object is a cell.

[0169] In another preferred embodiment, the method is non-therapeutic.

[0170] In another preferred embodiment, the method is non-therapeutic in vitro.

[0171] In a sixth aspect of the present invention, a method for treating or preventing a disease associated with free radicals is provided, wherein the method comprises the steps of:

[0172] A safe and effective amount of the heme derivative as described in the first aspect, the compound of formula IIa as described in the second aspect, and / or the composition as described in the third aspect is administered to a subject in need.

[0173] In another preferred embodiment, the subject is a human or a non-human mammal.

[0174] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0175] The inventors have conducted extensive and in-depth research. It was unexpectedly discovered that heme or its derivatives connected with a tripeptide chain and the middle amino acid residue is His have similar or even better catalase activity than heme or its analogs connected with only 2 peptides, and have better water solubility and are easier to make into medicines. In addition, since only a tripeptide is required, the difficulty of synthesis is greatly reduced, thereby providing a new catalase mimetic as shown in Ia. In addition, the inventors also unexpectedly discovered that when the peptide chain is connected to a specific site of heme or its analog, its catalase activity can be further improved. In addition, the inventors have also discovered a class of prodrugs that themselves also have certain catalase activity, and these prodrugs can be further hydrolyzed into compounds with excellent enzyme activity in vivo. Based on this, the inventors have completed the present invention.

[0176] the term

[0177] Herein, the heme derivative of the present invention, the heme derivative short peptide of the present invention, the catalase mimetic of the present invention and the compound of the present invention can be used interchangeably to refer to the heme derivative as described in the first aspect.

[0178] As used herein, "compounds of formula Ia", "heme derivatives of formula Ia" and "heme analogs shown in formula Ia" are used interchangeably and refer to compounds having a structure as shown in formula Ia in the first aspect. Similarly, "compounds of formula Ib-1", "heme derivatives of formula Ib-1" and "heme analogs shown in formula Ib-1" are used interchangeably; "compounds of formula Ib-2", "heme derivatives of formula Ib-2" and "heme analogs shown in formula Ib-2" are used interchangeably; "compounds of formula Ic", "heme derivatives of formula Ic" and "heme analogs shown in formula Ic" are used interchangeably.

[0179] As used herein, "halogen" refers to F, Cl, Br, and I. More preferably, the halogen atom is selected from F, Cl and Br.

[0180] As used herein, "C1-C6 alkyl" refers to a straight or branched alkyl group including 1-6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, or similar groups. Preferably, the alkyl group includes a straight or branched alkyl group of 1-4 carbon atoms (i.e., "C1-C4 alkyl"). The term "alkenyl" refers to an unsaturated alkyl group having one or more double bonds. Similarly, the term "alkynyl" refers to an unsaturated alkyl group having one or more triple bonds. Examples of such unsaturated alkyl groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologues and isomers.

[0181] As used herein, "C1-C6 alkoxy" includes straight or branched alkoxy groups of 1-6 carbon atoms. For example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, or similar groups. Similarly, the alkoxy group is a straight or branched alkoxy group including 1-4 carbon atoms, i.e., "C1-C4 alkoxy".

[0182] As used herein, "C3-C6 cycloalkyl" refers to a cycloalkyl group containing 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like.

[0183] As used herein, the term "heterocycloalkyl" or "heterocyclyl" refers to a saturated or unsaturated cycloalkyl containing one to five heteroatoms selected from N, O and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Heterocycloalkyl can be a monocyclic, bicyclic or polycyclic system. Non-limiting examples of heterocycloalkyl include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S, S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. Heterocycloalkyl can be connected to the rest of the molecule via a ring carbon or a heteroatom. For terms such as cycloalkylalkyl and heterocycloalkylalkyl, it is meant that the cycloalkyl or heterocycloalkyl is attached to the rest of the molecule through an alkyl or alkylene linker. For example, cyclobutylmethyl- is a cyclobutyl ring attached to a methylene linker to the rest of the molecule.

[0184] Unless otherwise stated, the term "aryl" represents a polyunsaturated (usually aromatic) hydrocarbon radical, which can be a monocyclic or polycyclic (maximum three rings) fused together or covalently attached. The term "heteroaryl" refers to an aryl (or ring) containing 1 to 5 heteroatomic atoms selected from N, O, and S, wherein nitrogen and sulfur atoms are optionally oxidized, and nitrogen atom is optionally quaternized. Heteroaryl can be attached to the remainder of the molecule by heteroatoms. The non-limiting example of aryl includes phenyl, naphthyl and biphenyl, and the non-limiting example of heteroaryl includes pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuranyl (isobenzo ] aryl, thiazolyl, pyrrolyl, thiazolyl, furyl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridine, benzothiazolyl, benzofuranyl, benzothiophenyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furanyl, thienyl and the like. Substituents for each of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.

[0185] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthio, aralkyl) includes both aryl and heteroaryl rings as defined above. Thus, the term "aralkyl" is meant to include those groups in which an aryl group is attached to an alkyl group that is attached to the rest of the molecule (e.g., benzyl, phenethyl, pyridylmethyl, etc.).

[0186] The term "alkylene" by itself or as part of another substituent refers to a divalent radical derived from an alkane, for example -CH2CH2CH2CH2-.

[0187] Unless otherwise indicated, the term "halo" or "halogen" by itself or as part of another substituent refers to a fluorine, chlorine, bromine, or iodine atom. In addition, terms such as "haloalkyl" are meant to include monohaloalkyl or polyhaloalkyl. For example, the term "C 1-4 "Haloalkyl" is meant to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0188] Unless otherwise defined, herein, amino acids include natural amino acids or non-natural amino acids, including D-type and / or L-type amino acids. Examples of amino acids include, but are not limited to, Ala (A), Arg (R), Asn (N), Asp (D), Cys (C), Gln (Q), Glu (E), Gly (G), His (H), Ile (I), Leu (L), Lys (K), Met (M), Phe (F), Pro (P), Ser (S), Thr (T), Trp (W), Tyr (Y), Val (V). Preferably, in this article, the amino acid is an amino acid selected from the group consisting of L-glycine (L-Gly), L-alanine (L-Ala), β-alanine (β-Ala), L-glutamic acid (L-Glu), L-aspartic acid (L-Asp), L-histidine (L-His), L-arginine (L-Arg), L-lysine (L-Lys), L-valine (L-Val), L-serine (L-Ser), and L-threonine (L-Thr).

[0189] As used herein, the term "amino acid residue" refers to a group formed by removing one H from the N-terminal -NH2 of an amino acid and removing -OH from the C-terminal -COOH.

[0190] Active ingredients

[0191] As used herein, the term "compound of the present invention" refers to a compound (heme derivative) of Formula I. The term also includes enantiomers, diastereomers or racemates, various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of Formula I, or prodrugs thereof.

[0192] In another preferred embodiment, the compound of the present invention is a heme derivative as shown in Formula Ia, Ib-1 or Ib-2, or Ic as described in the first aspect.

[0193] In another preferred embodiment, the compound of the present invention is the compound as shown in IIa as described in the second aspect, which is used as a prodrug of the compound of formula Ia.

[0194] In a specific embodiment, the present invention provides a heme derivative or a salt thereof as shown in Formula III,

[0195]

[0196]

[0197] in,

[0198] The chiral carbon atoms C* and C*2 are each independently S-type, R-type, or a combination thereof;

[0199] n = 0 or 1;

[0200] m = 0 or 1;

[0201] A - Selected from the group consisting of chloride ion, bromide ion, hydroxide ion;

[0202] X is selected from the group consisting of -CH2-, -N(R')-, -O-, -S-;

[0203] Y is selected from the following group: -CH2- or -CH2CH2-, -C(R')2, -N(R')-, -O-;

[0204] R1 and R2 are each independently selected from: hydrogen, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy; or R1 and R2 and the carbon atoms connected thereto together form a C3-C7 cycloalkyl or a 3- to 7-membered heterocyclic ring,

[0205] R3 is selected from: -C1-C18 alkoxy, -C1-C18 alkylamino, a polypeptide containing 1 to 6 alpha (α) or beta (β) amino acids (residues), wherein the C-terminal substituent of the polypeptide is selected from the following group: hydroxyl, amino, C1-C6 alkoxy, C1-C6 alkylamino.

[0206] R4 is selected from the group consisting of hydroxy, amino, methoxy, C1-C6 alkoxy, C1-C6 alkylamino;

[0207] R5 and R6 are each independently selected from: hydrogen, vinyl, formyl, C1-C4 substituted or unsubstituted alkyl, and the substituent is selected from methoxy, ethoxy, fluorine, chlorine, bromine, dimethylamino, amino or hydroxyl.

[0208] M is a metal of Group VIII, Group I, Group II, or Group VII in the Periodic Table (as a coordination metal), preferably iron, manganese, cobalt, nickel, or zinc.

[0209] In another preferred embodiment, in particular, R4 and X can form a ring through C1-C8 alkanes, alkenes, and alkynes containing heteroatoms, wherein the heteroatoms refer to N, O, and S.

[0210] In another preferred embodiment,

[0211] n=0, the chiral carbon atom C* is S-type;

[0212] m=0,

[0213] X is -NH- or -O-;

[0214] R1 is selected from: hydrogen, methyl, ethyl, isopropyl, or phenyl; R2 is a hydrogen atom,

[0215] R3 is selected from the group consisting of: amino, C1-C18 alkyl, hydroxyl, a polypeptide comprising 1 to 4 alpha or beta amino acids (residues); preferably, the amino acids (residues) are derived from: L-glycine, L-alanine, beta-alanine, L-glutamic acid, L-aspartic acid, L-histidine, L-arginine, L-lysine, L-valine, and / or L-serine;

[0216] M is selected from the group consisting of iron, manganese and zinc.

[0217] In another preferred embodiment,

[0218] n=1,m=0 or 1,A- is chloride ion or hydroxide ion;

[0219] Y is -CH2- or -NH-,

[0220] R1 is hydrogen or methyl, preferably hydrogen; R2 is hydrogen or methyl, preferably hydrogen;

[0221] R3 is selected from the group consisting of amino, hydroxyl, methoxy, and a polypeptide consisting of 1-4 alpha amino acids, preferably, the amino acid is selected from L-alanine, L-glutamic acid, L-aspartic acid, L-arginine, L-lysine, L-valine, and L-serine, and the C-terminal substituent is amino or hydroxyl;

[0222] M is selected from iron, manganese and zinc.

[0223] In another preferred embodiment,

[0224] R3 is selected from a polypeptide of 1 or 4 alpha amino acids, preferably, the amino acids are selected from L-alanine, L-glutamic acid, L-aspartic acid, L-arginine, L-lysine, L-valine, L-serine, and the C-terminal substituent is an amino group;

[0225] R4 is selected from the group consisting of: hydroxy, amino, methoxy, ethoxy, N,N-dimethyl (ie -N(CH3)2), preferably hydroxy.

[0226] In another preferred embodiment,

[0227] Each R5 is methyl; and each R6 is selected from: hydrogen, methyl, vinyl, ethyl, 1-methoxyethyl, 1-bromoethyl or formyl, preferably hydrogen.

[0228] or

[0229] Each R6 is methyl; and each R5 is selected from: hydrogen, methyl, vinyl, ethyl, 1-methoxyethyl, 1-bromoethyl or formyl, preferably hydrogen;

[0230] M is preferably iron or manganese.

[0231] In another preferred embodiment, the compound of the present invention is selected from the compounds in Table C

[0232] Table C

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240]

[0241]

[0242] The term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Acids suitable for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid. Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0243] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of the compound of the present invention with water.

[0244] Prodrug

[0245] The present invention also provides a prodrug of the heme derivative represented by Formula I (more preferably, the heme derivative represented by Formula Ia).

[0246] The term "prodrug" includes a substance which may be biologically active or inactive, and which undergoes metabolism or chemical reactions in the human body after being taken by an appropriate method to be converted into a compound of Formula I (such as Formula Ia), or a salt or solution composed of a compound of Formula I (such as Formula Ia). The prodrug includes (but is not limited to) carboxylate, carbonate, phosphate, nitrate, sulfate, sulfone, sulfoxide, amino compound, carbamate, azo compound, phosphoramide, glucoside, ether, acetal and the like forms of the compound.

[0247] In a specific embodiment, the present invention also provides a prodrug as shown in formula IIa. This type of prodrug can be recognized by enzymes in vivo to form a compound (tripeptide derivative of heme) as shown in formula Ia. Specifically, the prodrug as shown in formula II can be recognized by pancreatic amylase in the intestine to hydrolyze the peptide bond between the 3rd and 4th amino acid residues.

[0248] Pharmaceutical compositions and methods of administration

[0249] Since the compounds of the present invention have excellent peroxidase activity, they have excellent effects of catalyzing hydrogen peroxide, oxidizing phenolic and amine compounds and hydrocarbon oxidation products, and eliminating the toxicity of hydrogen peroxide and phenolic, amine, aldehyde, and benzene. Therefore, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and prodrugs, as well as pharmaceutical compositions containing the compounds of the present invention as the main active ingredients can be used to treat, prevent, and alleviate diseases caused by the accumulation of peroxides and / or free radicals. According to the prior art, the compounds of the present invention can be used to treat the following diseases: ischemia-reperfusion injury, diabetic complications, Oppenheimer's disease, etc.

[0250] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or its prodrug or its pharmacologically acceptable salt, and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention / dose, more preferably, contains 10-500 mg of the compound of the present invention / dose. Preferably, the "one dose" is a capsule or tablet.

[0251] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers Wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0252] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0253] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0254] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.

[0255] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

[0256] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0257] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0258] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0259] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0260] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0261] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0262] Preparation method

[0263] The preparation methods of the compounds of formula I (heme derivatives) or their prodrugs of the present invention are described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention belongs.

[0264] Typically, the method for preparing the compound of formula I (heme derivative) or its prodrug of the present invention comprises the steps of:

[0265] coupling fragment 1 and fragment 2 to obtain a compound of formula I (heme derivative) or a prodrug thereof;

[0266] Wherein, fragment 1 has a substituted or unsubstituted porphyrin ring structure that forms a planar coordination structure with the metal M (preferably, the fragment 1 is heme or a heme analogue);

[0267] Fragment 2 has a first coordination structure capable of further forming a stereo-coordination structure with the metal M (preferably, the fragment 2 is a peptide chain coupled to a resin or not coupled thereto).

[0268] In another preferred embodiment, fragment 1 has a structure selected from the following group:

[0269]

[0270] Wherein, the definitions of each group are as described above

[0271] In another preferred embodiment, fragment 2 has a structure selected from the following group, in each formula, the active group (such as -NH-, -NH2) can be optionally protected by a protecting group (such as Fmoc-) (such as -N(Fmoc)-, -NH(Fmoc)):

[0272] HX-(Y) n -C(R1)R2-C(O)NH-His-A a -R t ,

[0273] HX-(Y) n -C(R1)R2-C(O)NH-His-(A) o -R t ,

[0274] HX-(Y) n -C(R1)R2-C(O)NH-His-A a -(A') P -R t ,

[0275] HX-(Y) n -C(R1)R2-C(O)NH-His-A a -NH-resin,

[0276] HX-(Y) n -C(R1)R2-C(O)NH-His-(A) o -NH-resin,

[0277] HX-(Y) n -C(R1)R2-C(O)NH-His-A a -(A') P -NH-resin;

[0278] Wherein, each group is defined as above.

[0279] In a specific embodiment, the preparation method provided by the present invention comprises the steps of:

[0280] (1) synthesizing a peptide chain, for example, by solid phase peptide synthesis; and

[0281] (2) reacting the peptide chain synthesized in step (1) with a heme-like moiety, and optionally cleaving and removing the resin to obtain a compound of formula I.

[0282] In a specific embodiment, the compounds of the present invention can be synthesized according to the following method:

[0283] The preparation method of the compound of the present invention is synthesized by Fmoc solid phase peptide synthesis; specifically,

[0284] Raw materials and reagents: The peptide resin is Rink Amide MBHA resin. The amino acid derivatives are Fmoc-β-Ala-OH,

[0285] Fmoc-His(Trt)-OH, Fmoc-Thr(But)-OH, Fmoc-β-Thr(But)-OH, Fmoc-Val-OH, Fmoc-β-Val-OH, Fmoc-Glu(otBu)-OH, Fmoc-γ-Glu(otBu)-OH, Fmoc-Lys(Boc)-OH, etc.

[0286] The coupling reaction condensation agent is benzotriazole-1-oxy-tris(dimethylamino)phosphine hexafluorophosphate (BOP), 1-hydroxybenzotriazole

[0287] (HOBT), HATU, HBTU, HCTU, T3P;

[0288] The bases are N-methylmorpholine (NMM), triethylamine, and pyridine.

[0289] The deprotecting agent is 5-40% piperidine / DMF solution, 5-40% piperidine / dichloromethane solution.

[0290] The cleavage reagent was trifluoroacetic acid.

[0291] The resin was swollen with DMF / DCM for 30 minutes, then deprotected for 20 minutes, washed with DMF, and the amino acid derivative and condensing agent were added to the reactor, reacted at room temperature for 2 hours, washed, deprotected again, and the amino acid derivative and condensing agent were added, and the process was repeated until all amino acids were connected. Finally, heme and condensing agent were added, reacted at room temperature for 8 hours, and washed. The cleavage reagent was added to the reactor again, reacted at room temperature for 1 hour, and then filtered. The filtrate was precipitated in ether to obtain a crude product.

[0292] Purification of crude product: Purification was performed by high performance liquid chromatography. The eluent was trifluoroacetic acid aqueous solution and acetonitrile. The flow rate was 20 ml / min and the monitoring wavelength was 387 nm.

[0293] The main advantages of the present invention include:

[0294] (a) The compounds of the present invention are easy to synthesize;

[0295] (b) The compound of the present invention has excellent catalase activity and can effectively scavenge DPPH·free radicals.

[0296] (c) The compounds of the present invention have good in vitro stability and are not easily deteriorated.

[0297] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the unrecorded specific conditions in the following examples are usually carried out under normal conditions, according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts.

[0298] Example 1 Solid Phase Synthesis of D1 (Dh-β-Ala-His-Ala)

[0299]

[0300] Raw materials: Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Ala-OH coupling reaction condensation agent is benzotriazole-1-oxy-tris(dimethylamino)phosphine hexafluorophosphate (PyBOP), 1-hydroxybenzotriazole (HOBT), N-methylmorpholine (NMM). Deprotection agent is 20% piperidine / DMF solution. Cutting agent is trifluoroacetic acid.

[0301] Method 1:

[0302] 1. Weigh 0.1282 g of Rink Amide resin, put it into the treated reactor, add 10 mL of dichloromethane, shake it thoroughly to resuspend the resin, and let it stand at room temperature for 1 hour to fully swell the resin.

[0303] 2. Remove the residual liquid in the reactor under reduced pressure, add 5 ml of N, N-dimethylformamide to the reactor, plug the upper stopper, and shake it thoroughly for 1 min. Filter by suction, and wash with 5 ml of N, N-dimethylformamide 3 times. Add 5 ml of 20% piperidine / DMF solution to the reactor, shake it slowly, and shake it in a shaker for 30 min. Filter by suction under reduced pressure, wash with 5 ml of N, N-dimethylformamide 3 times, take a small amount of resin, and detect it with ninhydrin. The resin is dark blue, and the deprotection is considered complete.

[0304] 3. Add Fmoc-Ala-OH (93.40 mg, 3 eq); HOBt (40.53 mg, 3 eq); Py BOP: (156.12 mg, 3 eq) to the reactor, add 3 mL DMF to dissolve and mix with the resin, add NMM (66 μL, 3 eq) in DMF (8 mL) to the reactor, plug the upper stopper, and place it in a shaker for 4 hours. Filter under reduced pressure, and wash the resin 3 times with DMF (5 mL / time). Detect with ninhydrin. If and only if the heated solution is light yellow and the resin beads are almost colorless, the coupling is considered complete.

[0305] 4. Deprotect the resin after coupling with alanine, and replace Fmoc-Ala-OH (93.40 mg, 3 eq) with Fmoc-His(Trt)-OH (186 mg, 3 eq); repeat steps 2 and 3;

[0306] 5. Deprotect the resin after coupling with histidine, and replace Fmoc-Ala-OH (93.40 mg, 3 eq) with Fmoc-β-Ala-OH (93.40 mg, 3 eq); repeat steps 2 and 3;

[0307] 6. Deprotect the resin coupled with β-alanine and repeat step 2;

[0308] 7. Add hematin (45.2 mg, 0.8 eq); PyBOP (62.4 mg, 1.2 eq); HOBt (20.3 mg, 1.5 eq) to the reactor; add 3 mL of dichloromethane to mix and suspend the resin, add a dichloromethane (10 mL) solution of NMM (22 μL, 1 eq), place it in a shaker and shake for 10 hours, filter it under reduced pressure, the resin is black, wash the resin with dichloromethane 4 times, 10 mL each time, until the dichloromethane solution is colorless. Wash the resin again with anhydrous methanol 3 times, 10 mL each time, until the methanol solution is colorless. Dry the resin in vacuo.

[0309] 8. Add cleavage reagent to the reactor: trifluoroacetic acid: water: Tis = 95: 2.5: 2.5 (v / v / v), 6 mL, slowly shake until the resin is evenly dispersed, and place in a shaker to shake for 1.5 hours. Slowly drop the mixture into 30 mL of anhydrous ether precooled to -20°C, centrifuge at 4°C 6000 rpm for 5 minutes, and discard the supernatant. Wash the precipitate with ether (30 mL), centrifuge to remove the ether, repeat 3 times, and dry the precipitate to obtain 120 mg of crude product.

[0310] 9. The crude product was dissolved in water / methanol and purified using a C18, C8 or C3 reverse phase column. The eluent was a 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min and the detection wavelength was 214 nm. The main peak with a longer retention time was collected and freeze-dried to obtain 44 mg of the pure product.

[0311] Since these compounds contain iron coordination, HNMR cannot be detected, so these compounds are identified by ESI mass spectrometry. ESI-MS m / z = 842 [M] +

[0312] Example 2 Solid Phase Synthesis of D2 (Dh-β-Ala-His-Arg)

[0313]

[0314] Fmoc-Arg(Pbf)-OH was used to replace Fmoc-Ala-OH in Example 1, and the synthesis method was referred to Method 1 to prepare Compound D2. ESI-MS m / z=927[M] +

[0315] Example 3 Solid Phase Synthesis of D3 (Dh-β-Ala-His-Asp)

[0316]

[0317] Fmoc-Asp(OtBu)-OH was used to replace Fmoc-Ala-OH in Example 1, and the synthesis method was referred to Method 1 to prepare Compound D3. ESI-MS m / z=886[M] +

[0318] Example 4 Solid Phase Synthesis of D4 (Dh-β-Ala-His-Glu)

[0319]

[0320] Fmoc-Glu(OtBu)-OH was used to replace Fmoc-Ala-OH in Example 1, and the synthesis method was referred to Method 1 to prepare Compound D4. ESI-MS m / z=900[M] +

[0321] Example 5 Solid Phase Synthesis of D5 (Dh-β-Ala-His-Lys)

[0322]

[0323] Fmoc-Lys(Boc)-OH was used to replace Fmoc-Ala-OH in Example 1, and the synthesis method was referred to Method 1 to prepare Compound D5. ESI-MS m / z=899[M] +

[0324] Example 6 Solid Phase Synthesis of D6 (Dh-β-Ala-His-Pro)

[0325]

[0326] Fmoc-Pro-OH was used to replace Fmoc-Ala-OH in Example 1, and the synthesis method was referred to Method 1 to prepare Compound D6. ESI-MS m / z=868[M] +

[0327] Example 7 Solid Phase Synthesis of D7 (Dh-β-Ala-His-Thr)

[0328]

[0329] Fmoc-Thr(Boc)-OH was used to replace Fmoc-Ala-OH in Example 1, and the synthesis method was referred to Method 1 to prepare Compound D7. ESI-MS m / z=872[M] +

[0330] Example 8 Solid Phase Synthesis of D8 (Dh-Tle-His-Glu)

[0331]

[0332] Fmoc-β-Ala-OH in Example 4 was replaced by Fmoc-Tle-OH, and the synthesis method was similar to that of compound D4 to prepare compound D8. ESI-MS m / z=942[M] +

[0333] Example 9 Solid Phase Synthesis of D9 (Dh-Tle-His-Asp)

[0334]

[0335] Fmoc-β-Ala-OH in Example 3 was replaced by Fmoc-Tle-OH, and the synthesis method was similar to that of compound D3 to prepare compound D9. ESI-MS m / z=928[M] +

[0336] Example 10 Solid Phase Synthesis of D10 (Dh-Aib-His-Glu)

[0337]

[0338] Fmoc-Aib-OH was used to replace Fmoc-β-Ala-OH in Example 4, and the synthesis method was similar to that of compound D4 to prepare compound D10. ESI-MS m / z=914[M] +

[0339] Example 11 Solid Phase Synthesis of D11 (Dh-Aib-His-Asp)

[0340]

[0341] Fmoc-Aib-OH was used to replace Fmoc-β-Ala-OH in Example 3, and the synthesis method was referred to the synthesis of compound D3 to prepare compound D11. ESI-MS m / z=900[M] +

[0342] Example 12 Solid Phase Synthesis of D12 (Dh-Aib-His-Asp)

[0343]

[0344] Fmoc-Phg-OH was used to replace Fmoc-β-Ala-OH in Example 4, and the synthesis method was similar to that of compound D4 to prepare compound D12. ESI-MS m / z=962[M] +

[0345] Example 13 Solid Phase Synthesis of D13 (Dh-Abu-His-Asp)

[0346]

[0347] Fmoc-β-Ala-OH in Example 4 was replaced by Fmoc-Abu-OH, and the synthesis method was similar to that of compound D4 to prepare compound D13. ESI-MS m / z=914[M] +

[0348] Example 14 Solid Phase Synthesis of D14 (Dh-Ala-His-Asp)

[0349]

[0350] Fmoc-β-Ala-OH in Example 4 was replaced by Fmoc-Ala-OH, and the synthesis method was referred to the synthesis of compound D4 to prepare compound D14. ESI-MS m / z=900[M] +

[0351] Example 15: D15 (Dh-Ala-His-Glu(OMe)-OMe)

[0352] Synthesis route:

[0353]

[0354] (i) Synthesis of Compound 15-3

[0355] Compound 15-1 (commercially available) (2g, 7.83mmol) and EDCI (1.8g, 9.4mmol) were dissolved in 100mL dichloromethane, and 15-2 (1.37g, 7.83mmol) and triethylamine (1.03g, 10.2mmol) were added under stirring at 20°C, and stirring was continued for 5 hours. The reaction solution was extracted twice with 50ml 1N hydrochloric acid, once with 50mL saturated sodium bicarbonate, and once with 50mL saturated brine. The organic layer was evaporated to dryness to obtain solid 15-3 (3g, yield 92.8%).

[0356] H NMR (CDC13, 300MHz) δ8.51 (s, 1H), 7.35 (s, 1H), 5.04-4.99 (m, 1H), 4.22-4.18 (m, 1H), 3.65 (s, 3H ), 3.55 (s, 3H), 3.01 (d, J=3.6Hz, 2H), 2.40-2.32 (m, 2H), 2.30-2.25 (m, 2H), 1.44 (s, 9H).ESI-MS m / z=413[M+H] +

[0357] (ii) Synthesis of Compound 15-4

[0358] Compound 15-3 (3 g, 7.27 mmol) was dissolved in 30 mL of dichloromethane, and 15 mL of trifluoroacetic acid was added, and stirred at 20°C for 1 hour. The reaction solution was evaporated to dryness to obtain a light yellow oil 15-4. No purification was required and it was used directly in the next step. ESI-MS m / z = 313 [M+H] +

[0359] (iii) Synthesis of Compound 15-6

[0360] The product of the previous step was dissolved in 100 mL of dichloromethane, and 15-5 (1.38 g, 7.27 mmol), EDCI (1.67 g, 8.73 mmol), triethylamine (1.33 mL, 9.46 mmol) were added and stirred at 20°C for 6 hours. The reaction solution was extracted twice with 50 mL of 1N hydrochloric acid, once with 50 mL of saturated sodium bicarbonate, and once with 50 mL of saturated brine. The organic layer was evaporated to dryness, and the obtained oil was chromatographed on a silica gel column with dichloromethane / methanol = 10 / 1 to obtain a colorless oily product 15-6 (1.5 g, yield 42.6%).

[0361] H NMR (CDC13, 300MHz) δ8.52 (s, 1H), 7.34 (s, 1H), 5.05-4.98 (m, 1H), 4.25-4.20 (m, 1H), 3.64 (s, 3 H), 3.55-3.45 (m, 5H), 3.01 (d, J=3.6Hz, 2H), 2.50-2.32 (m, 4H), 2.30-2.25 (m, 2H), 1.43 (s, 9H).

[0362] ESI-MS m / z=484[M+H] +

[0363] Synthesis of compound 15-7

[0364] Compound 15-6 (1 g, 2.07 mmol) was dissolved in 20 mL of dichloromethane, 10 mL of trifluoroacetic acid was added, and the mixture was stirred at 20°C for 1 hour. The reaction solution was evaporated to dryness to obtain a light yellow oil 15-7. No purification was required and it was used directly in the next step. ESI-MS m / z = 384 [M+H] +

[0365] (iv) Synthesis of Compound D15

[0366] The product of the previous step was dissolved in 50 mL of dichloromethane, and hematin (1.17 g, 2.1 mmol), PyBOP (1.13 g, 2.2 mmol), HOBt (440 mg, 3.3 mmol) and N-methylmorpholine (330 mg, 3.3 mmol) were added. The mixture was stirred at 20°C for 10 hours. The reaction solution was directly evaporated to dryness. The obtained black solid was chromatographed on a silica gel column with dichloromethane / methanol = 10 / 1 to obtain a crude product D-15. The crude product was dissolved in water / methanol and purified using a C18 reverse phase column. The eluent was 0.1% TFA aqueous solution for phase A and acetonitrile for phase B. The flow rate was 20 ml / min, and the detection wavelength was 214 nm. The main peak was collected and freeze-dried to obtain 550 mg of a pure product. The yield was 28.6%. ESI-MS m / z = 929 [M] +

[0367] Example 16: D16 (Dh-Ala-His-Glu-OH)

[0368]

[0369] Compound D15 (100 mg, 0.1 mmol) was dissolved in 10 mL of methanol, sodium hydroxide (20 mg, 0.5 mmol) was added, and stirred at 20°C for 1 hour. The reaction solution was concentrated to 2 mL and purified using a C18 reverse phase column. The eluent was 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min, and the detection wavelength was 214 nm. The main peak was collected and freeze-dried to obtain 80 mg of a pure product. The yield was 82%. ESI-MS m / z = 901 [M] +

[0370] Example 17 Solid Phase Synthesis of D17 (Dh-Nle-His-Glu)

[0371]

[0372] Fmoc-β-Ala-OH in Example 4 was replaced by Fmoc-Nle-OH, and the synthesis method was similar to that of compound D4 to prepare compound D17. ESI-MS m / z=942[M] +

[0373] Embodiment 18

[0374] Solid Phase Synthesis of D18(Dh-Nle-His-Asp)

[0375]

[0376] Fmoc-β-Ala-OH in Example 3 was replaced by Fmoc-Nle-OH, and the synthesis method was similar to that of compound D3 to prepare compound D18. ESI-MS m / z=928[M] +

[0377] Example 19 Solid Phase Synthesis of D19 (Dh-Nva-His-Glu)

[0378]

[0379] Fmoc-β-Ala-OH in Example 4 was replaced by Fmoc-Nva-OH, and the synthesis method was similar to that of compound D4 to prepare compound D19. ESI-MS m / z=928[M] +

[0380] Example 20 Solid Phase Synthesis of D20 (Dh-Nva-His-Asp)

[0381]

[0382] Fmoc-β-Ala-OH in Example 3 was replaced by Fmoc-Nva-OH, and the synthesis method was similar to that of compound D3 to prepare compound D20. ESI-MS m / z=914[M] +

[0383] Example 21 Solid Phase Synthesis of D21 (Dh-β-Ala-His-Ala-Ala-Ala-Arg)

[0384]

[0385] Fmoc-Arg(pbf)-OH, Fmoc-Ala-OH, Fmoc-His(Trt)-OH, and Fmoc-β-Ala-OH were used as raw materials. The synthesis method was similar to that of compound D1. Compound D21 was prepared by peptide solid phase synthesis in the order of Arg, Ala, Ala, Ala, His, β-Ala, and heme. ESI-MS m / z=1225[M] +

[0386] Example 22 Solid Phase Synthesis of D22 (Dh-β-Ala-His-Arg-Ala-Ala-Arg)

[0387]

[0388] Fmoc-Arg(pbf)-OH, Fmoc-Ala-OH, Fmoc-His(Trt)-OH, and Fmoc-β-Ala-OH were used as raw materials. The synthesis method was similar to that of compound D1. Compound D22 was prepared by peptide solid phase synthesis in the order of Arg, Ala, Ala, Arg, His, β-Ala, and heme. ESI-MS m / z=1225[M] +

[0389] Example 23 Solid Phase Synthesis of D23 (Dh-β-Ala-His-Ala-Arg-Arg-Ala)

[0390]

[0391] Fmoc-Arg(pbf)-OH, Fmoc-Ala-OH, Fmoc-His(Trt)-OH, and Fmoc-β-Ala-OH were used as raw materials. The synthesis method was similar to that of compound D1. Compound D23 was prepared by peptide solid phase synthesis in the order of Ala, Arg, Arg, Ala, His, β-Ala, and heme. ESI-MS m / z=1225[M] +

[0392] Example 24 Solid Phase Synthesis of D24 (Dh-β-Ala-His-Ala-Arg-Ala-Arg)

[0393]

[0394] Fmoc-Arg(pbf)-OH, Fmoc-Ala-OH, Fmoc-His(Trt)-OH, and Fmoc-β-Ala-OH were used as raw materials. The synthesis method was similar to that of compound D1. Compound D24 was prepared by peptide solid phase synthesis in the order of Arg, Ala, Arg, Ala, His, β-Ala, and heme. ESI-MS m / z=1225[M] +

[0395] Example 25 Solid Phase Synthesis of D25 (Dh-β-Ala-His-Ala-Arg-Pro-Arg)

[0396]

[0397] Fmoc-Arg(pbf)-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-His(Trt)-OH, and Fmoc-β-Ala-OH were used as raw materials. The synthesis method was similar to that of compound D1. Compound D25 was prepared by peptide solid phase synthesis in the order of Arg, Pro, Arg, Ala, His, β-Ala, and heme. ESI-MS m / z=1251[M] +

[0398] Example 26 Solid Phase Synthesis of D26 (Dh-iso-β-Ala-His-Thr-Val-Glu-Lys)

[0399]

[0400] Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Val-OH, Fmoc-Glu(OtBu)-OH, and Fmoc-Lys(Boc)-OH were used as raw materials. The synthesis method was based on the synthesis of compound D1. The D26 and D27 mixtures were prepared by the synthesis method of acidic dissociation and deprotection according to the connection order of Lys, Glu, Val, Thr, His, β-Ala, and heme. The mixtures were purified by using a C18, C8, or C3 reverse phase column. The eluent was 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min and the detection wavelength was 387 nm. The first main peak with a shorter retention time was collected and lyophilized to obtain compound D26. ESI-MS m / z=1228[M] +

[0401] Example 27 Solid Phase Synthesis of D27 (Dh-β-Ala-His-Thr-Val-Glu-Lys)

[0402]

[0403] The second main peak with a longer retention time in Example 26 was collected and lyophilized to obtain compound D27. ESI-MS m / z=1228[M] +

[0404] Example 28 Solid Phase Synthesis of D28 (Heme-iso-β-Ala-His-Thr-Val-Glu-Lys)

[0405]

[0406] Heme was used to replace the hypoheme in Example 26, and the synthesis method was referred to the synthesis of D26 to prepare compound D28. ES1-MS m / z=1280[M] +

[0407] Example 29 Solid Phase Synthesis of D29 (Heme-β-Ala-His-Thr-Val-Glu-Lys)

[0408]

[0409] The first main peak with a longer retention time in Example 28 was collected and lyophilized to obtain compound D29. ESI-MS m / z=1280[M] +

[0410] Example 30 Solid Phase Synthesis of D30 (Heme-keto-β-Ala-His-Thr-Val-Glu-Lys)

[0411]

[0412] The heme in Example 26 was replaced by diketoheme, and the synthesis method was referred to the synthesis of D26 to prepare compound D30. ESI-MS m / z=1312[M] +

[0413] Example 31 Solid Phase Synthesis of D31 (Dh-DM-β-Ala-His-Thr-Val-Glu-Lys)

[0414]

[0415] Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Val-OH, Fmoc-Glu(OtBu)-OH, and Fmoc-Lys(Boc)-OH were used as raw materials. The synthesis method was based on the synthesis of compound D1. Lys, Glu, Val, Thr, His, β-Ala, hematin, and dimethylamine were connected in order, and then acidic dissociation and deprotection were used to prepare D31. ESI-MS m / z=1255[M] +

[0416] Example 32 Solid Phase Synthesis of D32 (Dh-Nva-His-Thr-Val-Glu-Lys)

[0417]

[0418] Fmoc-Nva-OH was used to replace Fmoc-β-Ala-OH in Example 26, and the synthesis method was referred to the synthesis of D26 / D27 to prepare compound D32. ESI-MS m / z=1256[M] +

[0419] Example 33 Solid Phase Synthesis of D33 (Dh-Ala-His-Thr-Val-Glu-Lys)

[0420]

[0421] Fmoc-Ala-OH was used to replace Fmoc-β-Ala-OH in Example 26, and the synthesis method was referred to the synthesis of D26 / D27 to prepare compound D33. ESI-MS m / z=1228[M] +

[0422] Solid Phase Synthesis of D34 (Dh-Nle-His-Thr-Val-Glu-Lys)

[0423]

[0424] Fmoc-β-Ala-OH in Example 26 was replaced by Fmoc-Nle-OH, and the synthesis method was referred to the synthesis of D26 / D27 to prepare compound D34. ESI-MS m / z=1270[M] +

[0425] Solid Phase Synthesis of D35 (Dh-Aib-His-Thr-Val-Glu-Lys)

[0426]

[0427] Fmoc-Aib-OH was used to replace Fmoc-β-Ala-OH in Example 26, and the synthesis method was referred to the synthesis of D26 / D27 to prepare compound D35. ESI-MS m / z=1242[M] +

[0428] Solid Phase Synthesis of D36 (Dh-Tle-His-Thr-Val-Glu-Lys)

[0429]

[0430] Fmoc-Tle-OH was used to replace Fmoc-β-Ala-OH in Example 26, and the synthesis method was referred to the synthesis of D26 / D27 to prepare compound D36. ESI-MS m / z=1270[M] +

[0431] Solid Phase Synthesis of D37 (Heme-iso-β-Ala-His-Glu)

[0432]

[0433] Heme was used to replace the hypoheme in Example 4. The synthesis method was based on the synthesis of D4. The compound was purified by using a C18 reverse phase column. The eluent was a 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min and the detection wavelength was 387 nm. The first main peak with a shorter retention time was collected and lyophilized to prepare compound D37. ESI-MS m / z=952[M] +

[0434] Example 38 Solid Phase Synthesis of D38 (Heme-β-Ala-His-Glu)

[0435]

[0436] The second main peak with a longer retention time in Example 37 was collected and lyophilized to obtain compound D38. ESI-MS m / z=952[M] +

[0437] Example 39 Solid Phase Synthesis of D39 (Heme-iso-Ala-His-Glu)

[0438]

[0439] Heme was used to replace the hypoheme in Example 3. The synthesis method was based on the synthesis of D3. The compound was purified by using a C18 reverse phase column. The eluent was 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min and the detection wavelength was 387 nm. The first main peak with a shorter retention time was collected and lyophilized to prepare compound D39. ESI-MS m / z=952[M] +

[0440] Example 40 Solid Phase Synthesis of D40 (Heme-Ala-His-Glu)

[0441]

[0442] The second main peak with a longer retention time in Example 39 was collected and lyophilized to obtain compound D40. ESI-MS m / z=952[M] +

[0443] Example 41 Solid Phase Synthesis of D41 (Heme-β-Ala-His)

[0444]

[0445] Using Fmoc-β-Ala-OH and Fmoc-His(Trt)-OH as raw materials, the synthesis method was similar to that of compound D1, and His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D41. ESI-MS m / z=823[M] +

[0446] Example 42 Solid Phase Synthesis of D42 (Mn-β-Ala-His-Thr-Val-Glu-Lys)

[0447]

[0448] Compound 42-1 was used to replace the hypoheme in Example 26, and the synthesis method was referred to the synthesis of D26 / D27 to prepare compound D42. ESI-MS m / z=1227[M] +

[0449] Example 43 Solid Phase Synthesis of D43 (Dh-iso-β-Ala-His-Thr)

[0450]

[0451] Fmoc-Ala-OH in Example 1 was replaced with Fmoc-Thr(Boc)-OH. The synthesis method was referenced to method 1. Purification was performed using a C18 reverse phase column. The eluent was a 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min. The detection wavelength was 387 nm. The first main peak with a shorter retention time was collected and freeze-dried to prepare compound D43. ESI-MS m / z=872[M] +

[0452] Example 44 Solid Phase Synthesis of D44 (Dh-iso-β-Ala-His-Asp)

[0453]

[0454] The second main peak with a longer retention time in Example 3 was collected and lyophilized to obtain compound D44. ESI-MS m / z=886[M] +

[0455] Example 45 Solid Phase Synthesis of D45 (Dh-iso-β-Ala-His-Glu)

[0456]

[0457] The second main peak with a longer retention time in Example 4 was collected and lyophilized to obtain compound D45. ESI-MS m / z=900[M] +

[0458] Example 46 Solid Phase Synthesis of D46 (Dh-iso-β-Ala-His-Lys)

[0459]

[0460] The second main peak with a longer retention time in Example 5 was collected and lyophilized to obtain compound D46. ESI-MS m / z=899[M] +

[0461] Example 47 Solid Phase Synthesis of D47 (Dh-iso-β-Ala-His-Pro)

[0462]

[0463] The second main peak with a longer retention time in Example 6 was collected and lyophilized to obtain compound D47. ESI-MS m / z=868[M] +

[0464] Example 48 Solid Phase Synthesis of D48 (Dh-iso-Ala-His-Glu)

[0465]

[0466] The second main peak with a longer retention time in Example 14 was collected and lyophilized to obtain compound D48. ESI-MS m / z=900[M] +

[0467] Example 49 Solid Phase Synthesis of D4 (Dh-β-Ala-D-His-Glu)

[0468]

[0469] Fmoc-D-His(Trt)-OH was used to replace Fmoc-His(Trt)-OH in Example 4. The synthesis method was referred to method 1 to prepare a mixture of compounds D49 and D50. The mixture was purified by using a C8 reverse phase column. The eluent was a 0.1% TFA aqueous solution in phase A and acetonitrile in phase B. The flow rate was 20 ml / min and the detection wavelength was 387 nm. The first main peak with a shorter retention time was collected and lyophilized to obtain compound D49. ESI-MS m / z=900[M] +

[0470] Example 50 Solid Phase Synthesis of D50 (Dh-iso-β-Ala-D-His-Glu)

[0471]

[0472] The second main peak with a longer retention time in Example 49 was collected and lyophilized to obtain compound D50. ESI-MS m / z=900[M] +

[0473] Example 51 Solid Phase Synthesis of D51 (Dh-β-Ala-His-Thr-Val)

[0474]

[0475] Using Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, and Fmoc-Val-OH as raw materials, the synthesis method was referenced to the synthesis of compound D1, and Val, Thr, His, β-Ala, and heme were connected in order, and then acidic dissociation and deprotection were used to prepare D51. ESI-MS m / z=791[M] +

[0476] Example 52 Solid Phase Synthesis of D52 (Dh-β-Ala-His-Thr-Glu)

[0477]

[0478] Using Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, and Fmoc-Glu(OtBu)-OH as raw materials, the synthesis method was referenced to the synthesis of compound D1, and Glu, Thr, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D52. ESI-MS m / z=1001[M] +

[0479] Example 53 Solid Phase Synthesis of D53 (Dh-β-Ala-His-Thr-Lys)

[0480]

[0481] Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, and Fmoc-Lys(Boc)-OH were used as raw materials. The synthesis method was similar to that of compound D1. Lys, Thr, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D53. ESI-MS m / z=1000[M] +

[0482] Example 54 Solid Phase Synthesis of D54 (Dh-β-Ala-His-Glu-Lys)

[0483]

[0484] Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Glu(OtBu)-OH, and Fmoc-Lys(Boc)-OH were used as raw materials. The synthesis method was similar to that of compound D1. Lys, Glu, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D54. ESI-MS m / z=1028[M] +

[0485] Example 55 Solid Phase Synthesis of D55 (Dh-β-Ala-His-Val-Glu)

[0486]

[0487] Using Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Val-OH, and Fmoc-Glu(OtBu)-OH as raw materials, the synthesis method was referenced to the synthesis of compound D1, and Glu, Val, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D55. ESI-MS m / z=999[M] +

[0488] Example 56 Solid Phase Synthesis of D56 (Dh-β-Ala-His-Val-Lys)

[0489]

[0490] Using Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Val-OH, and Fmoc-Lys(Boc)-OH as raw materials, the synthesis method was based on the synthesis of compound D1, and Lys, Val, His, β-Ala, and heme were connected in order, and then acidic dissociation and deprotection were used to prepare D56. ESI-MS m / z=998[M]+

[0491] Example 57 Solid Phase Synthesis of D57 (Dh-β-Ala-His-Thr-Val-Glu)

[0492]

[0493] Using Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Val-OH, and Fmoc-Glu(OtBu)-OH as raw materials, the synthesis method was referenced to the synthesis of compound D1, and Glu, Val, Thr, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D57. ESI-MS m / z=1100[M] +

[0494] Example 58 Solid Phase Synthesis of D58 (Dh-β-Ala-His-Thr-Glu-Lys)

[0495]

[0496] Use Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Glu(OtBu)-OH,

[0497] Fmoc-Lys(Boc)-OH was used as the raw material. The synthesis method was similar to that of compound D1. Lys, Glu, Thr, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D58. ESI-MS m / z=1129[M] +

[0498] Example 59 Solid Phase Synthesis of D59 (Dh-β-Ala-His-Thr-Val-Lys)

[0499]

[0500] Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Val-OH, and Fmoc-Lys(Boc)-OH were used as raw materials. The synthesis method was similar to that of compound D1. Lys, Val, Thr, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D59. ESI-MS m / z=1099[M] +

[0501] Example 60 Solid Phase Synthesis of D60 (Dh-β-Ala-His-Val-Glu-Lys)

[0502]

[0503] Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Val-OH, Fmoc-Glu(OtBu)-OH, and Fmoc-Lys(Boc)-OH were used as raw materials. The synthesis method was similar to that of compound D1. Lys, Glu, Val, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D60. ESI-MS m / z=1127[M] +

[0504] Example 61 Solid Phase Synthesis of D61 (Dh-β-Ala-His-Glu-Lys-Val)

[0505]

[0506] Use Fmoc-β-Ala-OH, Fmoc-His(Trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Lys(Boc)-OH,

[0507] Fmoc-Val-OH was used as the raw material. The synthesis method was similar to that of compound D1. Val, Lys, Glu, His, β-Ala, and heme were connected in the order of acidic dissociation and deprotection to prepare D61. ESI-MS m / z = 1127 [M] +

[0508] Example 62 Solid Phase Synthesis of D62 (Dh-iso-β-Ala-His-Thr-Glu)

[0509]

[0510] The second main peak with a longer retention time in Example 52 was collected and lyophilized to obtain compound D62. ESI-MS m / z=1001[M] +

[0511] Example 63 Solid Phase Synthesis of D63 (Dh-iso-β-Ala-His-Glu-Lys)

[0512]

[0513] The second main peak with a longer retention time in Example 54 was collected and lyophilized to obtain compound D64. ESI-MS m / z=1028[M] +

[0514] Example 64 Solid Phase Synthesis of D64 (Dh-iso-β-Ala-His-Thr-Val-Glu)

[0515]

[0516] The second main peak with a longer retention time in Example 57 was collected and lyophilized to obtain compound D64. ESI-MS m / z=1100[M] +

[0517] Example 65 Solid Phase Synthesis of D65 (Dh-iso-β-Ala-His-Thr-Glu-Lys)

[0518]

[0519] The second main peak with a longer retention time in Example 58 was collected and lyophilized to obtain compound D65. ESI-MS m / z=1129[M]

[0520] Test Example 1: Determination of the activity of peroxidase mimics

[0521] The peroxidase activity was determined by detecting the absorbance change of hydrogen peroxide at 240 nm using hydrogen peroxide and ascorbic acid as dual substrates. The experiment was conducted in a 37°C circulating water bath, with the reaction carried out in a 50mmol / L pH 7.0 phosphate buffer system, an ascorbic acid concentration of 0.5mmol / L, a D1-D42 concentration of 2μmol / L, and a hydrogen peroxide concentration of 1mmo / L.

[0522] Taking MP-11 as a positive reference, the structure of MP-11 is as follows:

[0523]

[0524] The amount of enzyme required to convert 1 mmol / L substrate into product per second is defined as one unit of enzyme activity.

[0525] Table 1 Peroxidase activity assay of compounds 1-42

[0526]

[0527]

[0528] It can be seen from the test results in Table 1 that the compound of the present invention has excellent peroxidase activity, which is comparable to that of the positive control MP-11.

[0529] Test Example 2 Determination of free radical activity in scavenging 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH)

[0530] DPPH· was prepared into a solution with a concentration of 0.1mmol / L using anhydrous ethanol, and compound 1-42 was prepared into a solution with a concentration of 0.2mM using ultrapure water. 500μl of DPPH· and 500μl of the compound to be tested were taken respectively, mixed, and allowed to stand in a light-proof and sealed environment at 25°C for 30min. The absorbance at 517nm was detected using a U2501 UV-visible spectrophotometer, and the scavenging activity of compound 1-42 against DPPH· free radicals was calculated.

[0531] Table 2 DPPH·free radical activity determination of compounds 1-42

[0532]

[0533] It can be seen from the test results in Table 2 that the compounds of the present invention have excellent DPPH· free radical scavenging activity.

[0534] Test Example 3 Cell activity assay

[0535] Mitochondria of living cells contain dehydrogenase. Dehydrogenase can reduce exogenous MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium odorant, trade name Thiazole Blue) to insoluble purple crystals of formazan and deposit them in cells. After dissolving it with dimethyl sulfone, the optical density value can reflect the amount of formazan and the strength of mitochondrial enzyme activity. PC-12 cells were inoculated in 96-well plates and cultured overnight at 37°C. The supernatant was discarded, and the cells were pretreated with LY294002 for 1 hour (the application of LY294002 aggravated the damage effect of ischemia and hypoxia on cells), 15μM of the test compound was added, 15μL of MTT was added to each well, and the culture was continued for 4 hours. The culture medium was aspirated, 150μl of DMSO was added to each well, and the mixture was shaken for 10 minutes. The absorbance value at 540nm was detected by a multifunctional microplate reader, and the cell proliferation was detected by MTT method.

[0536] Table 3 Determination of protective effect on oxidative damage of nerve cells

[0537]

[0538] From the results in Table 3, it can be seen that the survival rate of nerve cells in the test compound group is significantly better than that in the negative control group, indicating that such compounds have excellent protective effects on oxidative damage of nerve cells.

[0539] Test Example 4

[0540] In order to determine the optimal relative position of heme and histidine in the primary structure of the molecule, we used alanine as the first peptide chain and connected heme and histidine at both ends to synthesize a series of Dh-An-H derivatives of different lengths (as shown in the table below):

[0541]

[0542]

[0543] The above peptides were synthesized by solid phase synthesis using C 18 After purification by reverse phase column chromatography, the molecular weights of the compounds were determined by MS to be 699.1, 770.1, 841.2, 912.2, 983.2, 1054.3, and 1125.3, respectively, which were consistent with the theoretical values. The enzyme activity was determined by using Vc and H2O2 as substrates to determine the peroxidase activity, where the amount of enzyme required to oxidize 1 μg of ascorbic acid per minute was defined as one activity unit. The results are shown in Table 4.

[0544] Table 4 Peroxidase activity determination of Dh-An-H derivatives

[0545]

[0546] It can be seen from Table 4 that when there is an alanine between heme and histidine, that is, Dh-A1-H, the enzyme activity is the highest; increasing or decreasing the number of alanines between them will lead to a gradual decrease in enzyme activity.

[0547] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A heme derivative or a pharmaceutically acceptable salt thereof, characterized in that: The heme derivative is selected from the following table; 2. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (i) the heme derivative according to claim 1; and (ii) a pharmaceutically acceptable carrier.

3. A method for scavenging free radicals, comprising the steps of: The subject is incubated with the heme derivative of claim 1 to thereby clear free radicals in the subject; and the method is non-therapeutic in vitro.

4. The method according to claim 3, characterized in that The object is a cell.

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