Mitochondrial targeted photosensitizer compound and preparation method and application thereof

Abstract

The present invention belongs to the technical field of medical materials, and discloses a preparation method and application of a compound shown in formula I. The compound can be used as a mitochondrial targeted photosensitizer to achieve mitochondrial specific fluorescent staining, generate reactive oxygen species under light, and effectively kill cancer cells. It can be used for mitochondrial targeted photodynamic therapy in living cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical materials, and in particular relates to a mitochondrial targeted photosensitizer compound and a preparation method and application thereof. Background Art

[0002] Photodynamic therapy (PDT) is a targeted therapy that uses light, photosensitizers and oxygen molecules to selectively treat tumors and other diseases through photodynamic reactions. Excitation light, photosensitizers and tissue oxygen content are the three major elements of PDT, and none of them can be missing. The targeting of photosensitizers and the production of oxygen determine the efficacy of PDT.

[0003] Mitochondria are the "power plants" of cells and have important physiological functions. Mitochondria-targeted photosensitizers can generate reactive oxygen species under light, thereby destroying the structure of mitochondria and effectively improving the killing efficiency of cancer cells. Currently, there is little research on mitochondrial photosensitizers, so there is an urgent need to develop mitochondrial-targeted photosensitizers and then carry out application research in the biomedical field.

[0004] Existing photosensitizers usually do not have mitochondrial targeting, and the killing efficiency of tumor cells is low. In order to achieve spatiotemporally controllable mitochondrial targeted photodynamic therapy, it is necessary to prepare sensitive, highly targeted, and effective mitochondrial targeted photosensitizers. Summary of the invention

[0005] A compound of formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate or pharmaceutically acceptable salt:

[0006]

[0007] Among them, X - is selected from anions; n is selected from an integer of 1-20; p is selected from an integer of 0-3; q is selected from an integer of 0-4; t is selected from an integer of 0-5;

[0008] Each R 1 , R 2 , R 3 are the same or different and are independently selected from hydrogen, halogen, C 1-30 Alkyl, C 1-30 Alkoxy, C 6-30 Aryl, 5-20 membered heteroaryl.

[0009] According to an embodiment of the present invention, X - can be selected from halogen anions; n can be selected from integers of 1-12; p can be selected from integers of 0-1; q can be selected from integers of 0-2; t can be selected from integers of 0-2;

[0010] Each R1 , R 2 , R 3 may be the same or different and are independently selected from hydrogen, halogen, C 1-12 Alkyl, C 1-12 Alkoxy, C 6-14 Aryl, 5-14 membered heteroaryl.

[0011] According to an embodiment of the present invention, X - Can be selected from Cl - Br - ,I - ; n can be selected from 3, 4, 5, 6, 7 or 8; p is selected from 0 or 1; q is selected from 0, 1 or 2; t is 0;

[0012] Each R 1 , R 2 , R 3 may be the same or different and independently selected from hydrogen, F, Cl, Br, I.

[0013] According to an embodiment of the present invention, X - Br - ; n is 6; p is 0; q is selected from 0, 1 or 2; t is 0;

[0014] R 1 is hydrogen; each R 2 are the same or different and are independently selected from hydrogen, Cl or I; R 3 For hydrogen.

[0015] According to an embodiment of the present invention, the compound represented by formula I can be selected from the following structures:

[0016]

[0017]

[0018] The present invention also provides a method for preparing the compound represented by formula I, comprising the following steps:

[0019]

[0020] (1) Compound A reacts with compound B to obtain compound C;

[0021] (2) Compound C reacts with compound D to obtain a compound represented by formula I;

[0022] Among them, R 1 , R 2 , R 3 , X - , n, p, q, and t have the definitions described above.

[0023] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound represented by Formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate or pharmaceutically acceptable salt.

[0024] According to an embodiment of the present invention, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

[0025] The present invention also provides the use of at least one of the compound represented by Formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate or pharmaceutically acceptable salt as a photosensitizer, for example, as a mitochondrial targeting photosensitizer.

[0026] The present invention also provides the use of at least one of the compound represented by formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate or pharmaceutically acceptable salt in the preparation of photodynamic anti-tumor drugs.

[0027] According to an embodiment of the present invention, the drug may be a photosensitizing drug or a fluorescence-mediated drug; preferably, the drug may be targeted and enriched in the mitochondria of tumor cells.

[0028] According to an embodiment of the present invention, the tumor may be cancer, such as cervical cancer.

[0029] The present invention also provides the use of the compound represented by formula I, its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate or pharmaceutically acceptable salt in treating and / or preventing tumors.

[0030] The present invention also provides a method for treating and / or preventing tumor diseases, comprising administering to a patient a therapeutically or preventively effective amount of a compound of Formula I, or at least one of its racemates, stereoisomers, tautomers, isotope-labeled substances, solvates or pharmaceutically acceptable salts.

[0031] Beneficial Effects

[0032] Compared with the prior art, the present invention has the following advantages and effects:

[0033] The compound represented by formula I of the present invention can achieve specific fluorescence staining of mitochondria under illumination (intensity of illumination is 1 to 100 mW / cm 2 ) generates reactive oxygen species, which can effectively kill cancer cells.

[0034] The compound of the present invention can realize mitochondria-targeted photodynamic therapy in living cells under the condition of light irradiation, and has the advantages of strong cell entry ability and high photodynamic activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 Schematic diagram of compound Ⅰ-1 converting oxygen into active oxygen under ultraviolet light.

[0036] Figure 2 This is the H NMR spectrum of compound II-1 in Example 1.

[0037] Figure 3 This is the NMR carbon spectrum of compound II-1 in Example 1.

[0038] Figure 4 This is the H NMR spectrum of compound I-1 in Example 1.

[0039] Figure 5 This is the NMR carbon spectrum of compound I-1 in Example 1.

[0040] Figure 6 This is the H NMR spectrum of compound II-2 in Example 1.

[0041] Figure 7 This is the NMR carbon spectrum of compound II-2 in Example 1.

[0042] Figure 8 This is the H NMR spectrum of compound I-2 in Example 1.

[0043] Fig. 9 It is the NMR carbon spectrum of compound I-2 in Example 1.

[0044] Fig.10 This is the H NMR spectrum of compound II-3 in Example 1.

[0045] Fig.11 This is the NMR carbon spectrum of compound II-3 in Example 1.

[0046] Fig.12 This is the H NMR spectrum of compound I-3 in Example 1.

[0047] Fig.13 It is the NMR carbon spectrum of compound I-3 in Example 1.

[0048] Fig.14 In Example 2, 2',7'-dichlorodihydrofluorescein was used as an active oxygen indicator, and compound I-1 or I-2 (10 μM) was used as a photosensitizer. White light (100 mW cm -2 ) under irradiation, the fluorescence intensity of the indicator changes with the illumination time.

[0049] Fig.15 This is the co-staining image of compound Ⅰ-1 and MitoTracker Red in HeLa cells in Example 3.

[0050] Fig.16This is the co-staining image of compound Ⅰ-2 and MitoTracker Red in HeLa cells in Example 4.

[0051] Fig.17 The different concentrations of compound I-1 or I-2 in Example 5 were detected under white light (100 mW cm -2 ) Changes in cell survival rate under irradiation.

[0052] Definition and explanation of terms

[0053] Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. The definitions of groups and compound structures after such combinations and combinations should be understood to be within the scope of the specification and / or claims of this application.

[0054] Unless otherwise specified, the numerical ranges recorded in this specification and claims are equivalent to recording at least each specific integer value therein. For example, the numerical range "1-30" is equivalent to recording each integer value in the numerical range "1-30", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30.

[0055] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0056] The term "C 1-30 The term "alkyl" is understood to mean a linear or branched saturated monovalent hydrocarbon radical having 1 to 30 carbon atoms. For example, it means a linear or branched alkyl radical having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 1, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 carbon atoms. 1-12The term “alkyl” refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, or the like or isomers thereof.

[0057] The term "C 6-20 "Aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic (such as fused, bridged, spiro) or tricyclic hydrocarbon ring having 6 to 20 carbon atoms, which may be a single aromatic ring or a polyaromatic ring fused together, preferably "C 6-14 The term "C 6-14 The term "aryl" is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C 6-14 or a ring having 9 carbon atoms ("C9 aryl"), for example indanyl or indenyl, or a ring having 10 carbon atoms ("C 10 aryl), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C 13 aryl), such as fluorenyl, or a ring having 14 carbon atoms ("C 14 aryl”), such as anthracenyl. When the C 6-20 When the aryl group is substituted, it may be monosubstituted or polysubstituted. Also, there is no limitation on the substitution position, for example, it may be substituted at the ortho position, para position or meta position. Representative aryl groups include: phenyl, naphthyl, anthracenyl, pyrenyl.

[0058] The term "5-20 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic (e.g. fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, for example "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and, in each case, furthermore, may be benzo-fused. "Heteroaryl" also refers to a radical in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring. Representative heteroaryl groups include pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, thiazolyl, indolyl, azanaphthyl, azaanthryl, azapyrenyl.

[0059] In the present invention, "photosensitizer" refers to a class of molecules that effectively generate active oxygen under light irradiation. DETAILED DESCRIPTION

[0060] The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[0061] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0062] Example 1

[0063] Synthesis of compound II-1:

[0064]

[0065] Compound III-1 (85 mg, 0.5 mmol) and compound IV-1 (150 mg, 0.5 mmol) were dissolved in anhydrous ethanol (10 mL), the reaction solution was refluxed for 3 hours, the precipitate was filtered and washed three times with ethanol, and further vacuum dried to obtain compound II-1 (179 mg, yield 79%). 1H NMR (CDCl3, 400MHz): δ8.60(s,1H),8.56(s,1H),7.30–7.21(m,3H),6.96(d,J=8.8H z,1H),6.51(s,2H),4.00(t,J=6.4Hz,1H),3.43(t,J=6.4Hz,1H),1.90–1.51(m,8H); 13 C NMR(CDCl3,100MHz):164.7,164.0,162.1,161.8,158.1,134.0,132.7,131.1,124.3,118 .6,118.4,110.5,108.3,101.6,68.1,33.7,32.6,28.8,27.9,25.2; HRMS(ESI):m / z[M+H] + Calculated value C 20 H 23 BrClN2O3:453.0581; Exp. value:453.1001.

[0066] Synthesis of compound I-1:

[0067]

[0068] Compound II-1 (45.2 mg, 0.1 mmol) was dissolved in pyridine (10 mL), and the reaction solution was refluxed for 4 hours, then dried, and further recrystallized in a mixed solvent of n-hexane / ethyl acetate to obtain compound I-1 (47 mg, yield 88%). 1 HNMR (DMSO-d6, 400MHz): δ11.42 (s, 1H), 11.17 (s, 1H), 9.12 (d, J = 6.4Hz, 2H), 8.93 (s, 1H), 8.88(s,1H),8.61(t,J=7.6Hz,1H),8.17(t,J=6.4Hz,2H),7.74(s,1H),7.57(d,J=8.4Hz,2 H),7.00(d,J=8.8Hz,2H),6.55(d,J=8.4Hz,1H),6.51(s,1H),6.53(t,J=7.2Hz,2H),4.01( t,J=6.4Hz,2H),1.95(t,J=7.2Hz,2H),1.72(t,J=6.4Hz,2H),1.47–1.34(t,J=6.4Hz,4H); 13C NMR(DMSO-d6,100MHz):163.6,163.1,160.8,160.8,159.3,157.0,145.4,144.7,132.8,132.7,132.3 ,128.6,128.0,123.0,120.1,118.5,118.4,111.2,107.6,101.3,67.6,60.7,30.6,28.2,25.1,24.8.

[0069] Synthesis of compound II-2:

[0070]

[0071] Compound III-2 (148 mg, 0.5 mmol) and compound IV-1 (150 mg, 0.5 mmol) were dissolved in anhydrous ethanol (10 mL), the reaction solution was refluxed for 3 hours, the precipitate was filtered and washed three times with ethanol, and further vacuum dried to obtain compound II-2 (194 mg, yield 67%). 1 H NMR (CDCl3, 400MHz): δ8.58(s,1H),8.48(s,1H),7.79(s,1H),7.30–7.21(m,2H),6.53–6.50( m,2H),4.00(t,J=6.4Hz,1H),3.43(t,J=6.4Hz,1H),1.92–1.82(m,4H);1.51(t,J=6.4Hz,1H); 13 CNMR(CDCl3,100MHz):165.3,164.3,162.2,160.6,157.1,141.0,134.2,131.4 ,125.0,117.7,110.4,108.4,101.6,85.5,68.1,33.7,32.6,28.8,27.9,25.2.

[0072] Synthesis of compound I-2:

[0073]

[0074] Compound II-2 (58 mg, 0.1 mmol) was dissolved in pyridine (10 mL), and the reaction solution was refluxed for 4 hours, then dried by spin drying, and further recrystallized in a mixed solvent of n-hexane / ethyl acetate to obtain compound I-2 (51 mg, yield 77%). 1H NMR(DMSO-d6,400MHz): δ11.34(br s,2H),9.09(d,J=8.4Hz,2H),8.96(s,1H),8.84(s,1H),8.58(t,J=7.6H z,1H),8.15(t,J=6.4Hz,2H),7.87(s,1H),7.70(s,1H),7.55(d,J=8.4Hz ,1H),6.55–6.50(m,2H),4.60(t,J=7.2Hz,2H),3.99(t,J=6.0Hz,2H),1 .94(t,J=6.4Hz,2H),1.70(t,J=6.4Hz,2H),1.46–1.33(t,J=6.4Hz,4H); 13 C NMR(DMSO-d6,100MHz):163.5,163.2,161.3,160.9,145.4,144.7,139.9,132.4 ,130.7,128.0,118.4,111.2,107.6,101.4,67.6,60.7,30.6,28.1,25.1,24.8.

[0075] Synthesis of compound II-3:

[0076]

[0077] Compound III-3 (68 mg, 0.5 mmol) and compound IV-1 (150 mg, 0.5 mmol) were dissolved in anhydrous ethanol (10 mL), the reaction solution was refluxed for 3 hours, the precipitate was filtered and washed three times with ethanol, and further vacuum dried to obtain compound II-3 (176 mg, yield 81%). 1 H NMR (CDCl3, 400MHz): δ8.66(s,1H),8.61(s,1H),7.37–7.32(m,2H),7.24(d,J=8.4Hz,1H),7.02(d,J=8.0Hz,1H),6.95(t,J=7.2Hz,1 H),6.53–6.50(m,2H),4.00(t,J=6.4Hz,2H),3.43(t,J=6.4Hz,2H),1.90(t,J=6.4Hz,2H),1.81(t,J=6.0Hz,2H),1.51–1.50(m,4H); 13C NMR(CDCl3,100MHz):164.0,163.9,163.2,162.1,159.6,133.9,133.1,132.3,119.7,11 7.4,117.1,110.6,108.2,101.7,68.1,33.7,32.6,28.8,27.9,25.2; HRMS(ESI):m / z[M] + Calculated value C 20 H 23 BrN2O3:418.0892; Exp. value:418.1667.

[0078] Synthesis of compound I-3:

[0079]

[0080] Compound II-3 (42 mg, 0.1 mmol) was dissolved in pyridine (10 mL), the reaction solution was refluxed for 4 hours, then dried, and further recrystallized in a mixed solvent of n-hexane / ethyl acetate to obtain compound I-3 (46 mg, yield 92%). 1 H NMR (DMSO-d6, 400MHz): δ11.45 (s, 1H), 11.12 (s, 1H), 9.11 (d, J = 5.6Hz, 2H), 8.93 (s, 1H), 8.90(s,1H),8.60(t,J=6.4Hz,1H),7.65(d,J=7.2Hz,1H),7.54(d,J=8.4Hz,1H),7.37(t,J =7.2Hz,1H),6.95(d,J=8.8Hz,2H),6.53(d,J=8.8Hz,1H),6.49(s,1H),4.61(t,J=7.2Hz,2 H),3.99(t,J=6.8Hz,2H),1.70(t,J=7.2Hz,2H),1.44(t,J=6.8Hz,2H),1.45–1.34(m,4H); 13 C NMR(DMSO-d6,100MHz): δ163.0,162.9,161.6,160.7,158.4,145.4,144.7,132.9,132.8,130.7,128.0 ,119.5,118.2,116.4,111.2,107.5,101.3,67.6,60.6,30.5,28.1,25.0,24.8; HRMS(ESI):m / z[M+Na] + Calculated value C 25 H 28 BrN3NaO3:520.1212; Exp. value:520.8701.

[0081] Example 2

[0082] 2',7'-dichlorodihydrofluorescein was used as an active oxygen indicator to detect compound Ⅰ-1 or 1-2 (10 μM) in aqueous solution under white light (100 mW cm -2 ) ability to generate active oxygen under irradiation. The higher the fluorescence intensity over time, the more active oxygen is generated ( Fig.14 ), so the amount of active oxygen generated can be effectively controlled by controlling the light intensity and time.

[0083] Example 3

[0084] Compound Ⅰ-1 (10 μM) was added to the culture medium of HeLa cells and further co-stained with MitoTracker Red; Fig.15 From left to right in the figure are the cell fluorescence staining image of compound Ⅰ-1; the cell fluorescence staining image of MitoTracker Red; and the cell imaging overlay image.

[0085] Example 4

[0086] Compound I-2 (10 μM) was added to the culture medium of HeLa cells and further co-stained with MitoTracker Red; Fig.16 From left to right in the figure are the cell fluorescence staining image of compound Ⅰ-2; the cell fluorescence staining image of MitoTracker Red; and the cell imaging overlay image.

[0087] Example 5

[0088] Different concentrations of compound Ⅰ-1 or 1-2 were added to the HeLa cell culture medium and further exposed to white light (100 mWcm -2 ) Changes in cell survival rate under irradiation ( Fig.17 ).

[0089] The above is an explanation of the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A compound represented by formula I or a pharmaceutically acceptable salt thereof: in, X - Selected from Br - ; n is selected from 6; q is selected from 0, 1 or 2; Each R 2 are the same or different and are independently selected from hydrogen, F, Cl, Br, I.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: X - Br - ; n is 6; q is selected from 0, 1 or 2; Each R 2 are the same or different and are independently selected from hydrogen, Cl or I.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound shown in formula I is selected from the following structures:

4. A method for preparing the compound according to any one of claims 1 to 3, characterized in that: The following steps are involved: (1) Compound A reacts with compound B to obtain compound C; (2) Compound C reacts with compound D to obtain a compound represented by formula I; Among them, R 2 , X - , n, q have the definitions as described in any one of claims 1-3.

5. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds according to any one of claims 1 to 3 or pharmaceutically acceptable salts thereof.

6. Use of at least one of the compounds according to any one of claims 1 to 3 or pharmaceutically acceptable salts thereof in the preparation of a photosensitizer.

7. The use according to claim 6, characterized in that: The photosensitizer is a mitochondrial targeted photosensitizer.

8. Use of at least one of the compounds according to any one of claims 1 to 3 or pharmaceutically acceptable salts thereof in the preparation of photodynamic anti-tumor drugs.

9. The use according to claim 8, characterized in that: The drug is a photosensitizing drug.

10. The use according to claim 8, characterized in that: The drug is a fluorescence-mediated drug.

11. The use according to claim 8, characterized in that: The drug is targeted and enriched in the mitochondria of tumor cells.

12. The use according to claim 8, characterized in that: The tumor is a cancer.

13. The use according to claim 8, characterized in that: The tumor is cervical cancer.

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