CN114409663B - Zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugates targeting Mcl-1 enzyme and methods of making the same - Google Patents
Zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugates targeting Mcl-1 enzyme and methods of making the same Download PDFInfo
- Publication number
- CN114409663B CN114409663B CN202210103571.2A CN202210103571A CN114409663B CN 114409663 B CN114409663 B CN 114409663B CN 202210103571 A CN202210103571 A CN 202210103571A CN 114409663 B CN114409663 B CN 114409663B
- Authority
- CN
- China
- Prior art keywords
- chloro
- methylbenzo
- thiophene
- carboxylic acid
- dmf
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
- A61K41/0076—PDT with expanded (metallo)porphyrins, i.e. having more than 20 ring atoms, e.g. texaphyrins, sapphyrins, hexaphyrins, pentaphyrins, porphocyanines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/14—Drugs for genital or sexual disorders; Contraceptives for lactation disorders, e.g. galactorrhoea
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Gynecology & Obstetrics (AREA)
- Pregnancy & Childbirth (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Endocrinology (AREA)
- Reproductive Health (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugate targeting Mcl-1 enzyme and a preparation method thereof, belonging to the technical field of pharmaceutical chemistry. 3-nitrophthalonitrile is taken as an original raw material, 3- (4-carboxymethylester phenoxy) phthalonitrile is obtained through nucleophilic reaction, 1- (4-carboxyphenoxy) phthalocyanine is generated through cyclization reaction and nucleophilic reaction, and finally target product zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugate is generated through nucleophilic substitution reaction. The conjugate can enhance the killing effect on triple negative breast cancer cells and provides a new idea for improving photodynamic therapy.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical chemistry, and in particular relates to a zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugate targeting Mcl-1 enzyme and a preparation method thereof.
Background
Cancer is a major disease that jeopardizes human health. The existing treatment methods of cancers include chemotherapy, radiotherapy, surgical excision and photodynamic therapy, wherein photodynamic therapy has great potential due to selectivity, minimal invasion and low toxicity. The photosensitizer used in photodynamic therapy mainly comprises porphyrin and phthalocyanine, the phthalocyanine is a macrocyclic planar structure, and the targeting performance of the phthalocyanine is insufficient, so that the photodynamic therapy effect can be seriously reduced, and the improvement of the targeting performance of the phthalocyanine is an urgent problem to be solved.
Mcl-1 enzyme is highly expressed in some tumors, such as triple negative breast cancer, but is hardly expressed in normal tissues. Clinical data indicate that Mcl-1 enzyme is involved in the entire stage of the tumor, from its early stage to its metastasis and spread, and thus Mcl-1 enzyme is a potential target. 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid is an inhibitor of Mcl-1 kinase, which can be targeted to Mcl-1 enzymes.
Based on the important role of Mcl-1 enzyme in tumor development, the present invention proposes to combine the active domains of phthalocyanine and 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid, and to use the targeting of 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid to Mcl-1 enzyme to enhance the killing effect of the conjugate on triple negative breast cancer cells.
Disclosure of Invention
Aiming at the problem that phthalocyanine is easy to aggregate, the invention provides a zinc phthalocyanine-3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugate targeting Mcl-1 enzyme and a preparation method thereof. Targeting of 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid to Mcl-1 enzyme is utilized, and the targeting of phthalocyanine is enhanced by combining phthalocyanine and enzyme, so that the singlet oxygen yield is improved, and the treatment efficiency is further improved.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
zinc phthalocyanine-3-chloro-6-methylbenzo [ b ] targeting Mcl-1 enzyme]Thiophene-2-carboxylic acid conjugates having the chemical structural formula:
the preparation method of the zinc phthalocyanine-5-bromo-1-benzofuran-2-carboxylic acid conjugate targeting Mcl-1 enzyme comprises the following steps: 1) 3-Nitrophthalonitrile, methyl p-hydroxybenzoate and K 2 CO 3 Dissolving in DMF (dimethylformamide) according to a molar ratio of 1:1:2, reacting for 24-48 hours at normal temperature, adding a certain amount of water into the reaction liquid to precipitate white precipitate, filtering and drying to obtain a compound 1a, wherein the chemical structural formula is as follows:
wherein the volume ratio of water to DMF is 5:1;
2) Dissolving compound 1a and phthalonitrile in n-amyl alcohol, heating to 110-130 deg.C, adding zinc source or aluminium source, stirring uniformly, adding DUB (1, 8-diazabicyclo [ 5.4.0)]Undec-7-ene), reflux-reacting for 36-48 h, spin-drying n-amyl alcohol, adding excessive 20wt% sodium hydroxide aqueous solution, reflux-reacting for 24-48 h, spin-drying solution, adding 20wt% hydrochloric acid aqueous solution to adjust pH to about 3.0, filtering to obtain blue solid, taking the blue solid to pass through a silica gel column, and taking the volume ratio of EA (ethanol) -DMF as washing, wherein the volume ratio of EA (ethanol) -DMF is 100:1Eluting the stripping agent until no phthalocyanine exists, eluting by using the volume ratio of EA-DMF of 20:1, and collecting a target product 2a, wherein the chemical structural formula is as follows:
wherein the molar ratio of the compound 1a, phthalonitrile, DUB and zinc source is 1:7:8:4, and the zinc source is zinc chloride or zinc acetate;
3) 3-chloro-6-methylbenzo [ b ]]Thiophene-2-carboxylic acid, NHS (N-hydroxysuccinimide) and DCC (N, N-dicyclohexylcarbodiimide) are dissolved by DCM (dichloromethane), the mixture is dried after being reacted for 24 to 48 hours at normal temperature, the product is filtered by a neutral silica gel column, and a white solid 3a is collected by using the DCM as an eluent, wherein the chemical structural formula is as follows:
wherein 3-chloro-6-methylbenzo [ b ] is used]Thiophene-2-carboxylic acid, NHS and DCC in a molar ratio of 1:1:1;
4) Dissolving the product 2a, NHS, EDCI (carbodiimide) with THF (tetrahydrofuran), reacting for 48-72 h at normal temperature, adding diamine compound into the solution, refluxing for 1h, spin-drying the solution, adding water into a round bottom flask, shaking uniformly, pouring out the water, spin-drying, adding white solid 3a and DMF, reacting for 2h at 100-120 ℃, spin-drying, passing through a silica gel column, and using DCM-CH 3 Eluting with a volume ratio of OH of 100:1 to remove impurities, and then using DCM-CH 3 Eluting and purifying the OH with the volume ratio of 30:1, collecting the product, and repeatedly passing through the column for 2-3 times under the same eluting and purifying conditions to obtain a pure target product; wherein the molar ratio of the product 2a, NHS, EDCI, the hexamethylenediamine compound and the yellow solid 3a is 1:1:1:5:1.
The zinc phthalocyanine-3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate targeting Mcl-1 enzyme is applied to synthesis of photodynamic anti-tumor drugs.
The invention has the beneficial effects that:
1) The conjugate synthesized by the invention has stable structure, no isomer and easy synthesis;
2) The introduction of the 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid active domain in the invention can enhance the uptake of conjugates by tumor cells, and the 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid active domain can target Mcl-1 enzyme, so that the killing effect of the conjugates on triple-negative breast cancer cells is enhanced, and the effect of photodynamic therapy is enhanced.
Drawings
FIG. 1 is a process scheme for the synthesis of the phthalocyanine-3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate BC-Pc according to the present invention.
FIG. 2 is a process scheme for the synthesis of C6-Pc for the comparative example.
FIG. 3 is a graph showing the ultraviolet-visible absorption spectra of BC-Pc (A) and C6-Pc (B) synthesized in the examples in DMF.
FIG. 4 is a graph showing the toxicity of BC-Pc and C6-Pc under light conditions for MDA-MB-231 cells and HELF cells.
Detailed Description
In order to make the contents of the present invention more easily understood, the technical scheme of the present invention will be further described with reference to the specific embodiments, but the present invention is not limited thereto.
EXAMPLE 1 preparation of Zinc phthalocyanine 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate (BC-Pc)
3-nitrophthalonitrile is taken as an original raw material, 3- (4-carboxymethylphenoxy) phthalonitrile is obtained through nucleophilic reaction, 1- (4-carboxyphenoxy) zinc phthalocyanine is generated through cyclization reaction and alkaline hydrolysis reaction, and zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2 is generated through nucleophilic substitution reaction among 1- (4-carboxyphenoxy) zinc phthalocyanine, hexamethylenediamine, 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid active ester
The target product of the carboxylic acid conjugate (3-chloro-6-methyl-N- (6- (4- (phthalocyanine-1-yloxy) benzamido) hexyl) benzob-thiophen-2-carboxamide, BC-Pc). The synthetic route for the zinc phthalocyanine 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate is shown in FIG. 1.
The method comprises the following specific steps:
1) 2.00g (11.6 mmol) of 3-nitrophthalic acid methyl ester, 1.757g (11.6 mmol) of methylparaben, 3.202g (23.2 mmol) of K 2 CO 3 Dissolving in 20ml Dimethylformamide (DMF), and standing at room temperatureAfter 24 hours of reaction, 100ml of water was added to the reaction solution to precipitate a white precipitate, which was filtered and dried to obtain 2.5g of compound 1a in 78% yield, 1 H NMR(400MHz,Chloroform-d)δ8.13(d,J＝8.2Hz,2H),7.64(t,J＝8.2Hz,1H),7.54(d,J＝7.7Hz,1H),7.16(dd,J＝17.6,8.2Hz,3H),3.93(s,3H)；
the chemical structural formula of the compound 1a is:
2) 0.500g (1.8 mmol) of compound 1a and 1.448g (12.6 mmol) of phthalonitrile are dissolved in 50ml of n-amyl alcohol, after heating to 130 ℃, 1.318g (7.2 mmol) of zinc acetate is added, after stirring uniformly, 2.189g (14.4 mmol) of DBU is added, the solution turns green, then reflux reaction is carried out for 36h, then n-amyl alcohol is dried by spinning, 20ml of 20wt% sodium hydroxide aqueous solution is added, reflux reaction is carried out for 48h, 50ml of 20wt% hydrochloric acid aqueous solution is added into the solution after spinning, so as to adjust pH to 3.0, and blue solid is obtained by filtering; 300mg of the blue solid was applied to a silica gel column, washed with EA: DMF=100:1 (v/v) until no zinc phthalocyanine was present, eluted with EA: DMF=20:1 (v/v), and collected to give 200mg of the objective product 2a in 15% yield, 1 H NMR(400MHz,DMSO-d 6 )δ12.59(s,1H),9.12–8.76(m,6H),8.49(s,1H),8.26–7.69(m,10H),7.40(s,2H)。
the chemical structural formula of the target product 2a is as follows:
3) 0.633g (2.8 mmol) of 3-chloro-6-methylbenzo [ b]Thiophene-2-carboxylic acid, 0.327g (2.8 mmol) of N-hydroxysuccinimide (NHS), 0.576g (2.8 mmol) of N, N-Dicyclohexylcarbodiimide (DCC) were dissolved with 20 mM DCM, reacted at room temperature for 24h and then spun dry, eluted with DCM, passed through a neutral silica gel column, collected to give 0.6g of white solid 3a with a yield of 50%, 1 HNMR (500 MHz,) δ7.89 (d, j=8.4hz, 1H), 7.65-7.63 (m, 1H), 7.37-7.33 (m, 1H), 2.91 (s, 4H), 2.53 (s, 3H), white solid 3a has the chemical formula:
4) 100mg (0.14 mmol) of the target product 2a, 16.1mg (0.14 mmol) of N-hydroxysuccinimide, 26.74mg (0.14 mmol) of carbodiimide (EDCI) are dissolved in 20ml of THF (tetrahydrofuran) and reacted at room temperature for 48 hours, 81.2mg (0.70 mmol) of hexamethylenediamine is added to the solution, the reaction is carried out for 1 hour under reflux, the solution is dried by spinning, 50ml of water is added to a round bottom flask, a few shakes are carried out, the water is poured off, and then the mixture is dried by spinning, 45.22mg (0.14 mmol) of white solid 3a and 10ml of DMF are added to the flask and reacted for 2 hours at 100℃and after spinning the DMF is dried by spinning, DCM (dichloromethane): CH 3 OH (methanol) =100:1 (v/v) to remove impurities, and DCM: CH 3 OH=30:1 (v/v) elution and purification, collecting the product, repeating the column 3 times with the same eluting and purifying conditions, to obtain 20mg of pure product zinc phthalocyanine 3-chloro-6 methylbenzo [ b ]]Thiophene-2-carboxylic acid conjugate (BC-Pc) in a yield of 7%, 1 HNMR(500MHz,DMSO-d 6 )δ9.10–8.96(m,6H),8.54(d,J＝7.4Hz,1H),8.30(t,J＝5.7Hz,1H),8.22(t,J＝5.7Hz,1H),8.14–7.93(m,9H),7.81(t,J＝8.8Hz,1H),7.75(d,J＝0.6Hz,1H),7.60(d,J＝8.3Hz,1H),7.42–7.38(m,2H),7.27(dd,J＝8.3,0.9Hz,1H),3.18(tt,J＝14.2,7.2Hz,4H),2.37(s,3H),1.36–1.23(m,8H).HRMS(ESI):Calcd C 55 H 39 ClN 10 O 3 SZn[M+H] + :1019.1980,found:1019.1950.
the chemical structural formula of the zinc phthalocyanine 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate is as follows:
example 2
3-nitrophthalonitrile is taken as an original raw material, 3- (4-carboxymethylphenoxy) phthalonitrile is obtained through nucleophilic reaction, 1- (4-carboxyphenoxy) zinc phthalocyanine is generated through cyclization reaction and alkaline hydrolysis reaction, and a target product of BC-Pc is generated through nucleophilic substitution reaction among 1- (4-carboxyphenoxy) zinc phthalocyanine, hexamethylenediamine and 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid active ester.
The method comprises the following specific steps:
1) 2.00g (11.6 mmol) of 3-nitrophthalic acid methyl ester, 1.757g (11.6 mmol) of methylparaben, 3.202g (23.2 mmol) of K 2 CO 3 Dissolving in 20ml Dimethylformamide (DMF), reacting at normal temperature for 48h, adding 100ml water to the reaction solution to precipitate white precipitate, filtering and oven drying to obtain 2.5g compound 1a with yield of 80%, 1 H NMR(400MHz,Chloroform-d)δ8.13(d,J＝8.2Hz,2H),7.64(t,J＝8.2Hz,1H),7.54(d,J＝7.7Hz,1H),7.16(dd,J＝17.6,8.2Hz,3H),3.93(s,3H)；
the chemical structural formula of the compound 1a is:
2) 0.500g (1.8 mmol) of compound 1a and 1.448g (12.6 mmol) of phthalonitrile are dissolved in 50ml of n-amyl alcohol, after heating to 110 ℃, 0.982g (7.2 mmol) of zinc chloride is added, after stirring uniformly, 2.189g (14.4 mmol) of DBU is added, the solution turns green, then reflux reaction is carried out for 48 hours, then n-amyl alcohol is dried by spinning, 20ml of 20wt% sodium hydroxide aqueous solution is added, reflux reaction is carried out for 36 hours, 50ml of 20wt% hydrochloric acid aqueous solution is added into the solution after spinning, so as to adjust pH to 3.0, and blue solid is obtained by filtering; 300mg of blue solid was taken and applied to a silica gel column, washed with EA: DMF=100:1 (v/v) until no zinc phthalocyanine was present, eluted with EA: DMF=20:1 (v/v), collected to give 200mg of the desired product 2a in 16% yield, 1 HNMR(400MHz,DMSO-d 6 )δ12.59(s,1H),9.12–8.76(m,6H),8.49(s,1H),8.26–7.69(m,10H),7.40(s,2H)。
the chemical structural formula of the target product 2a is as follows:
3) 0.633g (2.8 mmol) of 3-chloro-6-methylbenzo [ b]Thiophene-2-carboxylic acid, 0.327g (2.8 mmol) of N-hydroxysuccinimide (NHS), 0.576g (2.8 mmol) of N, N-Dicyclohexylcarbodiimide (DCC) were dissolved with 20 mM DCM, reacted at room temperature for 48h and then spun dry, eluted with DCM, passed through a neutral silica gel column, collected to give 0.6g of white solid 3a with a yield of 55%, 1 HNMR(500MHz,)δ7.89(d,J＝8.4Hz,1H),7.65–7.63(m,1H),7.37–7.33 (m, 1H), 2.91 (s, 4H), 2.53 (s, 3H), white solid 3a has the chemical formula:
4) 100mg (0.14 mmol) of the target product 2a, 16.1mg (0.14 mmol) of N-hydroxysuccinimide, 26.74mg (0.14 mmol) of carbodiimide (EDCI) are dissolved in 20ml of THF (tetrahydrofuran), after a reaction at normal temperature of 72h, 81.2mg (0.70 mmol) of hexamethylenediamine is added to the solution, the reaction is carried out for 1 hour under reflux, the solution is dried by spinning, 50ml of water is added to a round-bottomed flask, after a few shakes, the water is poured off, and then dried by spinning, 45.22mg (0.14 mmol) of white solid 3a and 10ml of DMF are added to the flask, after spinning the DMF for 2h at 120℃the temperature, the mixture is dried by spinning with DCM (dichloromethane): CH 3 OH (methanol) =100:1 (v/v) to remove impurities, and DCM: CH 3 OH=30:1 (v/v) elution and purification, collecting the product, repeating the column 2 times with the same eluting and purifying conditions, to obtain 20mg of pure product zinc phthalocyanine 3-chloro-6 methylbenzo [ b ]]Thiophene-2-carboxylic acid conjugate (BC-Pc) in a yield of 9%, 1 HNMR(500MHz,DMSO-d 6 )δ9.10–8.96(m,6H),8.54(d,J＝7.4Hz,1H),8.30(t,J＝5.7Hz,1H),8.22(t,J＝5.7Hz,1H),8.14–7.93(m,9H),7.81(t,J＝8.8Hz,1H),7.75(d,J＝0.6Hz,1H),7.60(d,J＝8.3Hz,1H),7.42–7.38(m,2H),7.27(dd,J＝8.3,0.9Hz,1H),3.18(tt,J＝14.2,7.2Hz,4H),2.37(s,3H),1.36–1.23(m,8H).HRMS(ESI):Calcd C 55 H 39 ClN 10 O 3 SZn[M+H] + :1019.1980,found:1019.1950.
the chemical structural formula of the zinc phthalocyanine 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate is as follows:
example 3
3-nitrophthalonitrile is taken as an original raw material, 3- (4-carboxymethylphenoxy) phthalonitrile is obtained through nucleophilic reaction, 1- (4-carboxyphenoxy) zinc phthalocyanine is generated through cyclization reaction and alkaline hydrolysis reaction, and a target product of BC-Pc is generated through nucleophilic substitution reaction among 1- (4-carboxyphenoxy) zinc phthalocyanine, hexamethylenediamine and 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid active ester.
The method comprises the following specific steps:
1) 2.00g (11.6 mmol) of 3-nitrophthalic acid methyl ester, 1.757g (11.6 mmol) of methylparaben, 3.202g (23.2 mmol) of K 2 CO 3 Dissolving in 20ml Dimethylformamide (DMF), reacting at normal temperature for 36h, adding 100ml water to the reaction solution to precipitate white precipitate, filtering and oven drying to obtain 2.5g compound 1a with yield of 80%, 1 H NMR(400MHz,Chloroform-d)δ8.13(d,J＝8.2Hz,2H),7.64(t,J＝8.2Hz,1H),7.54(d,J＝7.7Hz,1H),7.16(dd,J＝17.6,8.2Hz,3H),3.93(s,3H)；
the chemical structural formula of the compound 1a is:
2) 0.500g (1.8 mmol) of compound 1a and 1.448g (12.6 mmol) of phthalonitrile are dissolved in 50ml of n-amyl alcohol, after heating to 120 ℃, 1.318g (7.2 mmol) of zinc acetate is added, after stirring uniformly, 2.189g (14.4 mmol) of DBU is added, the solution turns green, then reflux reaction is carried out for 40h, then n-amyl alcohol is dried by spinning, 20ml of 20wt% sodium hydroxide aqueous solution is added, reflux reaction is carried out for 36h, 50ml of 20wt% hydrochloric acid aqueous solution is added into the solution after spinning, so as to adjust pH to 3.0, and blue solid is obtained by filtering; 300mg of blue solid was taken and applied to a silica gel column, washed with EA: DMF=100:1 (v/v) until no zinc phthalocyanine was present, eluted with EA: DMF=20:1 (v/v), collected to give 200mg of the desired product 2a in 16% yield, 1 HNMR(400MHz,DMSO-d 6 )δ12.59(s,1H),9.12–8.76(m,6H),8.49(s,1H),8.26–7.69(m,10H),7.40(s,2H)。
the chemical structural formula of the target product 2a is as follows:
3) 0.633g (2.8 mmol) of 3-chloro-6-methylbenzo [ b]Thiophene-2-carboxylic acid, 0.327g (2.8 mmol) of N-hydroxysuccinimide (NHS), 0.576g (2.8 mmol) of N, N-Dicyclohexylcarbodiimide (DCC) were dissolved in 20 mM DCC, reacted at room temperature for 36hSpin-drying, passing through neutral silica gel column with DCM as eluent, collecting 0.6g of white solid 3a with yield of 55%, 1 HNMR (500 MHz,) δ7.89 (d, j=8.4hz, 1H), 7.65-7.63 (m, 1H), 7.37-7.33 (m, 1H), 2.91 (s, 4H), 2.53 (s, 3H), white solid 3a has the chemical formula:
4) 100mg (0.14 mmol) of the target product 2a, 16.1mg (0.14 mmol) of N-hydroxysuccinimide, 26.74mg (0.14 mmol) of carbodiimide (EDCI) are dissolved in 20ml of THF (tetrahydrofuran), after reaction at normal temperature for 60h, 81.2mg (0.70 mmol) of hexamethylenediamine is added to the solution, the reaction is carried out for 1 hour under reflux, the solution is dried by spinning, 50ml of water is added to a round-bottomed flask, after a few shakes, the water is poured off, and then dried by spinning, 45.22mg (0.14 mmol) of white solid 3a and 10ml of DMF are added to the flask, after spinning the DMF for 2h at 110℃the temperature, the mixture is dried by spinning with DCM (dichloromethane): CH 3 OH (methanol) =100:1 (v/v) to remove impurities, and DCM: CH 3 OH=30:1 (v/v) elution and purification, collecting the product, repeating the column 3 times with the same eluting and purifying conditions, to obtain 20mg of pure product zinc phthalocyanine 3-chloro-6 methylbenzo [ b ]]Thiophene-2-carboxylic acid conjugate (BC-Pc) in a yield of 9%, 1 HNMR(500MHz,DMSO-d 6 )δ9.10–8.96(m,6H),8.54(d,J＝7.4Hz,1H),8.30(t,J＝5.7Hz,1H),8.22(t,J＝5.7Hz,1H),8.14–7.93(m,9H),7.81(t,J＝8.8Hz,1H),7.75(d,J＝0.6Hz,1H),7.60(d,J＝8.3Hz,1H),7.42–7.38(m,2H),7.27(dd,J＝8.3,0.9Hz,1H),3.18(tt,J＝14.2,7.2Hz,4H),2.37(s,3H),1.36–1.23(m,8H).HRMS(ESI):Calcd C 55 H 39 ClN 10 O 3 SZn[M+H] + :1019.1980,found:1019.1950.
the chemical structural formula of the zinc phthalocyanine 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid conjugate is as follows:
comparative example Synthesis of N-hexyl-4- (Zinc phthalocyanine-1-oxy) benzamide
The process route for N-hexyl-4- (zinc phthalocyanine-1-oxy) benzamide (N-hexyl-4- (phthalocyanine-1-yloxy) benzamide, C6-Pc) is shown in FIG. 2. The method comprises the following specific steps:
100mg (0.14 mmol) of 2a, 16.1mg (0.14 mmol) of N-hydroxysuccinimide and 26.74mg (0.14 mmol) of EDCI are dissolved in 10ml of THF and reacted at normal temperature for 48h, after which 14.14mg (0.14 mmol) of N-hexylamine are added to the solution and reacted at reflux for 1 hour, the solution is spun dry with DCM: CH 3 OH=110:1 (v/v) eluting to remove impurities, followed by DCM: CH 3 OH=45:1 (v/v) elution and purification, collecting the product, repeating the column 3 times with the same eluting and purifying conditions, to obtain 30mg of purer product C6-Pc with a yield of 30%, 1 HNMR(400MHz,Chloroform-d)δ9.15(d,J＝44.7Hz,6H),8.79(s,1H),8.31–7.70(m,10H),7.63(s,2H),3.54(s,2H),1.40(s,9H),0.98(s,3H)，HRMS(ESI):Calcd C 45 H 33 N 9 O 2 Zn[M+H] + 796.2121, found:796.2083, C6-Pc has the following chemical structural formula:
application example 1 cytotoxicity test
The MTT method is also called a colorimetric method, and is a method for detecting the survival state of cells. Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT into formazan insoluble in water and deposit in cells, and dead cells cannot reduce MTT into formazan. Dimethyl sulfoxide can dissolve formazan in cells, and the wavelength value of the formazan at 570nm is measured by an enzyme-labeled instrument, so that the number of living cells can be indirectly reflected. The amount of MTT crystals formed is proportional to the number of cells over a range of cell numbers.
Selecting cells MDA-MB-231 and HELF with good growth state under a microscope, when the Cell density is about 90%, digesting with 1ml of pancreatin (containing EDTA) for 3 minutes (wherein Hela cells are digested for 1 minute), then adding 2ml of culture medium to stop digestion, blowing uniformly, counting by a Cell counting plate, diluting the Cell solution to 6 ten thousand cells/ml by using the DMEM culture medium, adding 100 mu L of the Cell culture medium solution into each well of a 96-well plate, setting 6 compound wells for each concentration of BF-Pc and C6-Pc medicines, setting a Cell blank group and a solvent blank group (6 compound wells for each group), and placing the 96-well plate into a 37 ℃ incubator for 24 hours.
Adding the medicine: BC-Pc was diluted to 200.0, 100.0, 50.0, 25.0, 12.5, 6.2, 3.1. Mu.M with 5vol% castor oil (CEL) in DMSO, C6-Pc to 25, 12.5, 6.2, 3.1, 1.6, 0.8, 0.4. Mu.M respectively with 5vol% castor oil in DMSO, then the 7 concentrations of drug were diluted 100-fold with DMEM medium, old medium in 96-well plates was aspirated with a lance, 100-fold dilution of 5vol% CEL in DMSO was added to the cell control, the solvent control was added to DMFM medium, the remaining drug control was added to the above-prepared solutions, and the 96-well plates were placed in the incubator for 24 hours.
Phototoxicity: taking out the 96-well plate, carefully sucking out the medicine-containing culture medium in the wells by using a gun, washing each well with physiological saline for 3 times to remove medicines which are not taken in, finally adding 100 mu L of the culture medium into each well, irradiating for 2min by using an LED lamp with the wavelength of 670nm, and then putting the irradiated plate back into the incubator.
Test OD value: mu.l of 5mg/ml MTT solution was added to each well of the 96-well plate, the 96-well plate was placed in an incubator at 37℃for 4 hours, then the solution of each well was sucked out with a discharge gun, 100. Mu.l of DMSO solution was added to each well again, and the mixture was shaken for 30 minutes with a shaker, and absorbance at 570nm was measured with a microplate reader.
And (3) calculating: the viability of the cells was calculated using the absorbance measured, as follows:
cell viability (%) = (sample group absorbance-solvent blank group absorbance)/(cell blank group absorbance-solvent blank group absorbance) ×100.
FIG. 3 is a graph showing the ultraviolet-visible absorption spectra of BC-Pc (A) and C6-Pc (B) in DMF, and it can be seen from the third graph that BC-Pc and C6-Pc are both substantially in a non-aggregated state in DMF.
FIG. 4 is a graph showing the toxicity of BC-Pc and C6-Pc under light conditions for MDA-MB-231 cells and HELF cells. From the figure, it can be seen that the introduction of 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid can enhance the toxic effect of BC-Pc on tumor cells.
The above description is only of the preferred embodiments of the present invention, and all the equivalent structures or equivalent processes or direct or indirect application in other related technical fields are included in the scope of the present invention.
Claims (1)
1. Use of a zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugate targeting Mcl-1 enzyme in the synthesis of photodynamic anti-tumour drugs, characterized in that: the chemical structural formula is as follows:
the preparation method comprises the following steps:
1) 3-Nitrophthalonitrile, methyl p-hydroxybenzoate and K 2 CO 3 Dissolving in DMF, reacting for 24-48 h at normal temperature, adding a certain amount of water into the reaction liquid to precipitate white precipitate, filtering and drying to obtain a compound 1a, wherein the chemical structural formula is as follows:
2) Dissolving a compound 1a and phthalonitrile in n-amyl alcohol, heating to 110-130 ℃, adding a zinc source, stirring uniformly, adding DBU, carrying out reflux reaction for 36-48 h, spin-drying n-amyl alcohol, adding an excessive 20wt% sodium hydroxide aqueous solution, carrying out reflux reaction for 24-48 h, adding a 20wt% hydrochloric acid aqueous solution to adjust pH to 3.0 after spin-drying the solution, filtering to obtain a blue solid, taking the blue solid to pass through a silica gel column, washing until no phthalocyanine exists by taking the volume ratio of EA-DMF as an eluent, eluting by taking the volume ratio of EA-DMF as 20:1, and collecting a target product 2a, wherein the chemical structural formula is:
3) Dissolving 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid, NHS and DCC with DCM, performing a reaction at normal temperature for 24-48 hours, performing spin-drying, passing the product through a neutral silica gel column, and collecting a white solid 3a by taking DCM as an eluent, wherein the chemical structural formula is as follows:
4) Dissolving a product 2a, NHS, EDCI by using THF, reacting for 48-72 h at normal temperature, adding a hexamethylenediamine compound into the solution, refluxing for 1h, spin-drying the solution, adding water into a round-bottomed flask, shaking uniformly, pouring out the water, spin-drying, adding a white solid 3a and DMF, reacting for 2h at 100-120 ℃, spin-drying, passing through a silica gel column, and using DCM-CH 3 Eluting with a volume ratio of OH of 100:1 to remove impurities, and then using DCM-CH 3 Eluting with the volume ratio of OH of 30:1, collecting the product, repeatedly passing through the column for 2-3 times under the same eluting and purifying conditions, and obtaining a pure target product;
3-nitrophthalonitrile, methyl p-hydroxybenzoate and K used in step 1) 2 CO 3 The molar ratio of water to DMF is 1:1:2, the volume ratio of water to DMF is 5:1;
the molar ratio of the compound 1a, phthalonitrile, DBU and zinc source in the step 2) is 1:7:8:4; the zinc source is zinc chloride or zinc acetate;
the molar ratio of 3-chloro-6-methylbenzo [ b ] thiophene-2-carboxylic acid, NHS and DCC used in the step 3) is 1:1:1;
the molar ratio of product 2a, NHS, EDCI, hexamethylenediamine, white solid 3a used in step 4) was 1:1:1:5:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210103571.2A CN114409663B (en) | 2022-01-28 | 2022-01-28 | Zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugates targeting Mcl-1 enzyme and methods of making the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210103571.2A CN114409663B (en) | 2022-01-28 | 2022-01-28 | Zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugates targeting Mcl-1 enzyme and methods of making the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114409663A CN114409663A (en) | 2022-04-29 |
| CN114409663B true CN114409663B (en) | 2023-05-30 |
Family
ID=81278327
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210103571.2A Active CN114409663B (en) | 2022-01-28 | 2022-01-28 | Zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugates targeting Mcl-1 enzyme and methods of making the same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114409663B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115160326A (en) * | 2022-07-30 | 2022-10-11 | 福州大学 | Phthalocyanine complex of targeted IDO enzyme and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5346670A (en) * | 1986-12-24 | 1994-09-13 | British Technology Group U.S.A. Inc. | Phthalocyanine and tetrabenztriazaporphyrin reagents |
| CN103073553A (en) * | 2013-01-25 | 2013-05-01 | 山东大学 | Water-soluble naphthalocyanine base compound, preparation method and application of compound as photosensitizer |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030119894A1 (en) * | 2001-07-20 | 2003-06-26 | Gemin X Biotechnologies Inc. | Methods for treatment of cancer or neoplastic disease and for inhibiting growth of cancer cells and neoplastic cells |
| WO2012086857A1 (en) * | 2010-12-21 | 2012-06-28 | (주)파낙스이엠 | Combination body for photodynamic diagnosis or therapy and a production method therefor |
| JP7055742B6 (en) * | 2015-08-18 | 2022-06-07 | ラクテン・メディカル，インコーポレイテッド | Phthalocyanine pigment conjugate and its preservation method |
| CN106432180A (en) * | 2016-09-07 | 2017-02-22 | 华东师范大学 | Compound containing benzo[b]thiophene structure, and derivative, preparation method and application |
| CN112574248A (en) * | 2020-12-10 | 2021-03-30 | 福建师范大学 | Preparation method of di- ((3-chlorobenzothiophene-2-ester group) hexafluorophenoxy) axial substituted silicon phthalocyanine |
| CN113943304B (en) * | 2021-10-21 | 2024-01-30 | 福州大学 | Pim-1 kinase targeted phthalocyanine-5-bromo-1-benzofuran-2-carboxylic acid complex, and preparation method and application thereof |
-
2022
- 2022-01-28 CN CN202210103571.2A patent/CN114409663B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5346670A (en) * | 1986-12-24 | 1994-09-13 | British Technology Group U.S.A. Inc. | Phthalocyanine and tetrabenztriazaporphyrin reagents |
| CN103073553A (en) * | 2013-01-25 | 2013-05-01 | 山东大学 | Water-soluble naphthalocyanine base compound, preparation method and application of compound as photosensitizer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114409663A (en) | 2022-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113943304B (en) | Pim-1 kinase targeted phthalocyanine-5-bromo-1-benzofuran-2-carboxylic acid complex, and preparation method and application thereof | |
| CN112209940A (en) | COX-2 enzyme-targeted phthalocyanine-indometacin complex and preparation method and application thereof | |
| CN107375929B (en) | Photosensitizer and derivatives and application thereof | |
| CN109575061B (en) | Water-soluble anticancer photosensitizer and preparation and application thereof | |
| CN109796483B (en) | Water-soluble cationic photosensitizer and preparation and application thereof | |
| CN114409663B (en) | Zinc phthalocyanine 3-chloro-6 methylbenzo [ b ] thiophene-2-carboxylic acid conjugates targeting Mcl-1 enzyme and methods of making the same | |
| CN104861039B (en) | A kind of phthalocyanine compound, preparation method and the application as single, double photon fluorescence probe in cancer targeting and mitochondrial markers | |
| CN103755713A (en) | Octa-sulfonic phthalocyanine, and preparation method and application thereof | |
| CN106946868B (en) | Nitric oxide donator type coumarin derivative, preparation method and medical usage | |
| CN111704646A (en) | Steroid compound and preparation method and application thereof | |
| CN110372754A (en) | A kind of novel metal complex of iridium and its preparation method and application | |
| CN110577526A (en) | Salt of bromodomain structural protein inhibitor and preparation method and application thereof | |
| CN115160326A (en) | Phthalocyanine complex of targeted IDO enzyme and preparation method and application thereof | |
| CN114685348B (en) | Near-infrared cyanine photosensitizer with AIE (AIE) property and preparation method and application thereof | |
| CN103073553B (en) | Water-soluble naphthalocyanine base compound, preparation method and application of compound as photosensitizer | |
| CN113278036B (en) | Phenothiazine-containing iridium complex and preparation method and application thereof | |
| CN106243114B (en) | Molecular targeted azepine aromatic rings axial substituted phthalocyanine complex and preparation method | |
| CN111808031B (en) | Ferulic acid derivative and preparation method and application thereof | |
| CN114835706B (en) | N≡N ligand and application thereof | |
| CN115947742A (en) | Imidazotetrazine group monosubstituted phthalocyanine conjugate and preparation method and application thereof | |
| CN114057777B (en) | Beta-carboline derivative and preparation method and application thereof | |
| CN114773361B (en) | N-heteronile erythronium salt compound and preparation method and application thereof | |
| CN116271024A (en) | Intracellular self-assembled vascular targeting photosensitizer and preparation method and application thereof | |
| CN113004264B (en) | Aggregation-induced emission photosensitizer, preparation method and application | |
| CN117964630A (en) | NO type porphyrin-ferulic acid derivative, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |