CN112480419B - Cisplatin carrier with MMP-2 response and preparation method thereof, cisplatin complex and preparation method thereof - Google Patents
Cisplatin carrier with MMP-2 response and preparation method thereof, cisplatin complex and preparation method thereof Download PDFInfo
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Abstract
The invention provides a cisplatin carrier with MMP-2 response and a preparation method thereof, and a cisplatin complex and a preparation method thereof. The cisplatin carrier provided by the invention has a structure shown in a formula I, a specific branched chain-thiol polyethylene glycol-maleimide polypeptide is grafted on a side chain of polyamino acid, the cisplatin carrier can be used as a cisplatin shell, has certain shielding capability and MMP-2 responsiveness, can be enriched at a tumor tissue part by enhancing a permeation-retention (EPR) effect, can prevent the toxicity of cisplatin in other normal environments of a human body, can quickly respond and automatically remove under the stimulation of active oxygen over-expressed at a tumor prevention part, and releases the cisplatin to play a role in a tumor part. Therefore, it can effectively avoid sudden release and nonspecific interaction in the blood circulation system after intravenous injection, thereby reducing toxicity.
Description
Technical Field
The invention relates to the technical field of high-molecular drug carriers, in particular to a cisplatin carrier with MMP-2 response and a preparation method thereof, and a cisplatin complex and a preparation method thereof.
Background
Tumors have become one of the most serious diseases threatening human health. The cancer treatment means commonly used in clinic include chemotherapy, radiotherapy, surgery and the like. Among them, chemotherapy is among the most common and important therapeutic approaches. However, clinically used antitumor drugs have many defects in application, such as: poor water solubility and stability, and great toxic and side effects of the medicine. In order to solve the problems, the medicament can be combined with a medicament carrier to improve the water solubility and the stability of the medicament and achieve the controlled release of the medicament, thereby reducing the toxic and side effects of the medicament on normal tissues and fully exerting the efficacy of the medicament.
Cisplatin (cis-diamminedichloroplatinum, abbreviated as CDDP) is a widely used anti-tumor drug, and was first discovered in 1965 by B.Rosenborg et al. Cisplatin has the characteristics of wide anticancer spectrum, strong action, synergistic action with various antitumor drugs, no cross drug resistance and the like, so cisplatin is also one of the most commonly used drugs in combined chemotherapy. However, cisplatin also has the problems of large toxic and side effects, short half-life in blood and the like, and particularly has kidney toxicity, so that the dosage of cisplatin is limited, and the application effectiveness of cisplatin is greatly limited.
Aiming at the problem, a Kataoka Research group of Tokyo university in Japan and a Chen academic Research group of Changchun institute of Chinese academy of sciences complex and carry cisplatin by using polyglutamic acid polymer as a carrier, and the prepared polymer-cisplatin nano-micelle obviously reduces the toxicity of cisplatin and can realize passive accumulation at tumor sites (N.Nishiyama, et al.cancer Research,63(2003)9877 one-shot 8983; Chinese patent 201210382696. X).
Although various cisplatin analogs have been developed to reduce toxic side effects and improve therapeutic effects, they still have the problems of short half-life in blood, significant nonspecific effects, and the like, which are common to small-molecule platinum drugs, and cause toxicity in the hematopoietic system, digestive system, and nervous system.
Disclosure of Invention
In view of the above, the present invention aims to provide a cisplatin carrier having MMP-2 response and a preparation method thereof, and a cisplatin complex and a preparation method thereof. The cisplatin carrier and the cisplatin complex provided by the invention can effectively avoid sudden release and nonspecific interaction in a blood circulation system after intravenous injection, thereby reducing toxicity.
The invention provides a cisplatin carrier with MMP-2 response, which has a structure shown in a formula I:
wherein:
40≤n≤120；
120≤x≤160，b＝0，5≤y≤15，1≤a≤y，
or
120≤x≤160，1≤b≤x，y＝a＝0。
Preferably, the first and second liquid crystal materials are,
140≤x≤155，b＝0，8≤y≤12，1≤a≤y，
or
140≤x≤155，1≤b≤x，y＝a＝0。
The invention also provides a preparation method of the cisplatin carrier with MMP-2 response in the technical scheme, which comprises the following steps:
A) mixing N-hexylamine, N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride and gamma benzyl glutamic acid-N-cyclic carboxylic anhydride for reaction to obtain polyamino acid nanoparticles;
B) dissolving the polyamino acid nanoparticles in halogenated acetic acid, and carrying out acidolysis in the presence of hydrogen bromide and acetic acid to remove carbobenzoxy groups, thereby obtaining a compound shown in a formula IV;
C) under the action of a catalyst, a functionalized nano shell compound and a compound shown in a formula IV are mixed and react to obtain a cisplatin carrier with MMP-2 response shown in the formula I;
the functionalized nano shell compound is a compound shown in a formula II and/or a compound shown in a formula III;
wherein:
40≤n≤120；
120≤x≤160，5≤y≤15，1≤a≤y，
or
120≤x≤160，y＝a＝0。
Preferably, in the step A), the reaction temperature is 15-50 ℃ and the reaction time is 2-7 days.
Preferably, in the step C), the reaction temperature is 10-30 ℃ and the reaction time is 1-3 h;
the catalyst is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine.
The invention also provides a cis-platinum complex which is formed by matching cis-diamminedichloroplatinum with a carrier;
the carrier is the cisplatin carrier with MMP-2 response in the technical scheme or the cisplatin carrier with MMP-2 response prepared by the preparation method in the technical scheme.
The invention also provides a preparation method of the cisplatin complex in the technical scheme, which comprises the following steps:
mixing the carrier and cis-diammine platinum dichloride for reaction to form a cis-platinum complex;
the carrier is the cisplatin carrier with MMP-2 response in the technical scheme or the cisplatin carrier with MMP-2 response prepared by the preparation method in the technical scheme.
Preferably, the reaction is carried out under the condition that the pH value is 8-10;
the reaction temperature is 30-45 ℃, and the reaction time is 2-5 days.
Preferably, the molar ratio of the glutamic acid unit to the cis-diamminedichloroplatinum in the carrier is (1-10) to 1.
Preferably, the reaction is carried out in solvent water.
According to the carrier with the structure of the formula I, a specific branched chain-thiol polyethylene glycol-maleimide polypeptide is grafted on the side chain of polyamino acid, the carrier can be used as a shell of cisplatin, has certain shielding capability and MMP-2 responsiveness, is enriched at a tumor tissue part through enhancing the osmotic retention (EPR) effect, can prevent the toxicity of the cisplatin in other normal environments of a human body, can quickly respond and automatically remove the cisplatin under the stimulation of active oxygen over-expressed at a tumor prevention part, and releases the cisplatin to play a role in a tumor part. Therefore, it can effectively avoid sudden release and nonspecific interaction in the blood circulation system after intravenous injection, thereby reducing toxicity.
Experimental results show that the cis-diammine-platinum dichloride complex has the characteristics of slow release capability and accelerated release under MMP-2 response.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a nuclear magnetic spectrum of N (. epsilon.) -benzyloxycarbonyl-L-lysine-N-cyclic lactam prepared in example 4;
FIG. 2 is a nuclear magnetic spectrum of gamma-benzylglutamic acid-N-cyclic anhydride prepared in example 5;
FIG. 3 is a nuclear magnetic spectrum of poly (L-lysine-co- γ -glutamic acid) of formula IV prepared in example 13;
FIG. 4 is a nuclear magnetic spectrum of cisplatin carrier nanoparticle of formula I prepared in example 25;
FIG. 5 is a graph showing the release profiles of cisplatin complexes prepared in accordance with the present invention under various conditions;
FIG. 6 is a graph showing the effect of MTT in the antitumor test in example 79.
Detailed Description
The invention provides a cisplatin carrier with MMP-2 response, which is characterized by having a structure shown in a formula I:
wherein:
40≤n≤120；
120≤x≤160，b＝0，5≤y≤15，1≤a≤y，
or
120≤x≤160，1≤b≤x，y＝a＝0。
In the structure of the formula I, n is more than or equal to 40 and less than or equal to 120; the number average molecular weight of the corresponding polyethylene glycol unit is preferably 2000-5000.
In the present invention, when y ≠ 0 or y ≠ 0, the structure of formula I is divided into two cases:
x is more than or equal to 120 and less than or equal to 160, b is 0, y is more than or equal to 5 and less than or equal to 15, and a is more than or equal to 1 and less than or equal to y, namely the compound of the formula I has a structure shown in the formula I-1:
among them, it is preferable that x is 140. ltoreq. x.ltoreq.155, y is 8. ltoreq. y.ltoreq.12, and a is 1. ltoreq. y.
Or
X is more than or equal to 120 and less than or equal to 160, b is more than or equal to 1 and less than or equal to x, and y is 0, namely the compound of the formula I has a structure shown in the formula I-2:
among them, it is preferable that 140. ltoreq. x.ltoreq.155 and 1. ltoreq. b.ltoreq.x.
According to the carrier with the structure of the formula I, a specific branched chain-thiol polyethylene glycol-maleimide polypeptide is grafted on the side chain of polyamino acid, the carrier can be used as a shell of cisplatin, has certain shielding capability and MMP-2 responsiveness, is enriched at a tumor tissue part through enhancing the osmotic retention (EPR) effect, can prevent the toxicity of the cisplatin in other normal environments of a human body, can quickly respond and automatically remove the cisplatin under the stimulation of active oxygen over-expressed at a tumor prevention part, and releases the cisplatin to play a role in a tumor part. Therefore, it can effectively avoid sudden release and nonspecific interaction in the blood circulation system after intravenous injection, thereby reducing toxicity.
The invention also provides a preparation method of the cisplatin carrier with MMP-2 response in the technical scheme, which is characterized by comprising the following steps:
A) mixing N-hexylamine, N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride and gamma benzyl glutamic acid-N-cyclic carboxylic anhydride for reaction to obtain polyamino acid nanoparticles;
B) dissolving the polyamino acid nanoparticles in halogenated acetic acid, and carrying out acidolysis in the presence of hydrobromic acid and acetic acid to remove carbobenzoxy to obtain a compound shown in a formula IV;
C) under the action of a catalyst, a functionalized nano shell compound and a compound shown in a formula IV are mixed and react to obtain a cisplatin carrier with MMP-2 response shown in the formula I;
the functionalized nano shell compound is a compound shown in a formula II and/or a compound shown in a formula III;
wherein:
40≤n≤120；
120≤x≤160，5≤y≤15，1≤a≤y，
or
120≤x≤160，y＝a＝0。
With respect to step a): n-hexylamine, N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride and gamma-benzylglutamic acid-N-cyclic carboxylic anhydride are mixed and react to obtain the polyamino acid nanoparticles.
In the present invention, the n-hexylamine structure is represented by the following formula a, and the source thereof is not particularly limited, and may be a general commercially available product.
In the present invention, the N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride (also known as "N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride" or "N6-benzyloxycarbonyl-L-lysine cyclic anhydride") has the structure shown in the following formula b, and the source thereof is not particularly limited, and it can be a commercially available product or prepared by a conventional preparation method known to those skilled in the art.
The N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride can be prepared by the following preparation method: carrying out condensation reaction on N (epsilon) -carbobenzoxy-L-lysine and bis (trichloromethyl) carbonate to obtain N (epsilon) -carbobenzoxy-L-lysine-N-cyclic internal anhydride.
Wherein, the specific process is preferably as follows: n (epsilon) -benzyloxycarbonyl-L-lysine and bis (trichloromethyl) carbonate were mixed in an organic solvent, heated, and subjected to a condensation reaction under anhydrous conditions. In the present invention, the molar ratio of N (epsilon) -benzyloxycarbonyl-L-lysine to bis (trichloromethyl) carbonate is preferably 1: 0.1 to 1.2, more preferably 1: 0.3 to 1, and most preferably 1: 0.5 to 0.8. The organic solvent is preferably tetrahydrofuran. The volume of the organic solvent and the mass ratio of the N (epsilon) -benzyloxycarbonyl-L-lysine are preferably (8-12) mL: 1g, and more preferably 10 mL: 1 g. In the invention, the mixing temperature is preferably 10-40 ℃, more preferably 15-35 ℃, and most preferably 20-30 ℃. The condensation reaction temperature is preferably 30-80 ℃, more preferably 35-70 ℃, most preferably 40-60 ℃, and the condensation reaction time is preferably 0.1-5 hours, more preferably 0.15-3 hours, and most preferably 0.2-2 hours.
After the above condensation reaction, it is preferable to precipitate the obtained reaction product with petroleum ether, separate the obtained precipitate, and then wash, recrystallize and dry the obtained separated product to obtain N (e) -benzyloxycarbonyl-L-lysine-N-cyclic lactam. The method of washing, recrystallization and drying in the present invention is not particularly limited, and washing, recrystallization and drying means known to those skilled in the art may be used.
In the present invention, the structure of the gamma-benzylglutamic acid-N-carboxyanhydride (also called "glutamic acid 5-benzyl ester N-carboxyanhydride") is represented by the following formula c, and the source thereof is not particularly limited, and it may be a general commercially available product or prepared according to a preparation method known to those skilled in the art.
The gamma-benzyl glutamic acid-N-ring carboxylic anhydride can be prepared by the following preparation method: carrying out condensation reaction on the gamma-benzyl glutamic acid and bis (trichloromethyl) carbonate to obtain the gamma-benzyl glutamic acid-N-cyclic internal anhydride.
Wherein, the specific process is preferably as follows: firstly, mixing the gamma-benzyl glutamic acid and the bis (trichloromethyl) carbonate in an organic solvent, then heating, and carrying out condensation reaction under the anhydrous condition. In the present invention, the molar ratio of the gamma-benzylglutamic acid to the bis (trichloromethyl) carbonate is preferably 1: 0.1 to 1.2, more preferably 1: 0.3 to 1, and most preferably 1: 0.5 to 0.8. The organic solvent is preferably tetrahydrofuran. The volume of the organic solvent and the mass ratio of the gamma-benzylglutamic acid are preferably (8-12) mL to 1g, and more preferably 10mL to 1 g. In the invention, the mixing temperature is preferably 10-40 ℃, more preferably 15-35 ℃, and most preferably 20-30 ℃. The condensation reaction temperature is preferably 30-80 ℃, more preferably 35-70 ℃, most preferably 40-60 ℃, and the condensation reaction time is preferably 0.1-5 hours, more preferably 0.15-3 hours, and most preferably 0.2-2 hours.
After the above condensation reaction, it is preferable to precipitate the obtained reaction product with petroleum ether, separate the obtained precipitate, and then wash, recrystallize and dry the obtained separated product to obtain γ -benzylglutamic acid-N-cyclic carboxylic anhydride. The method of washing, recrystallization and drying in the present invention is not particularly limited, and washing, recrystallization and drying means known to those skilled in the art may be used.
According to the invention, after three raw materials of N-hexylamine, N-benzyloxycarbonyl-L-lysine-N-endocyclic carboxylic anhydride and gamma-benzylglutamic acid-N-endocyclic carboxylic anhydride are obtained, the three raw materials are preferably dissolved in an organic solvent and then react under the condition of protective gas to obtain the polyamino acid nanoparticles.
In the invention, the dosage ratio of N-hexylamine, N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride and gamma-benzylglutamic acid-N-cyclic carboxylic anhydride is divided into two cases: (1) the final product corresponds to formula I-1 above: the molar ratio is preferably 1: 5-15: 120-160, more preferably 1: 8-12: 140-155, even more preferably 1: 9-11: 150-155, and most preferably 1: 10: 150. (2) The final product corresponds to formula I-2 above: the molar ratio is preferably 1: 0 to (120-160), more preferably 1: 0 to (140-155), even more preferably 1: 0 to (150-155), and most preferably 1: 0: 150.
In the present invention, the organic solvent is preferably N, N-Dimethylformamide (DMF). The preferable dosage ratio of the organic solvent to the n-hexylamine is (1-2) mL: 1.0 mg.
In the present invention, the kind of the protective gas is not particularly limited, and may be a conventional inert gas known to those skilled in the art, such as nitrogen, helium, argon, or the like. The pressure of the gas is not particularly limited, and the gas can be obtained at normal pressure.
In the invention, the reaction temperature is preferably 15-50 ℃, more preferably 20-40 ℃, and most preferably 25-35 ℃. The reaction time is preferably 2 to 7 days, more preferably 3 to 5 days, and most preferably 3 days.
In the present invention, after the reaction, it is preferable to further include: and pouring the obtained reaction liquid into anhydrous ether, carrying out suction filtration to obtain a solid, and then carrying out vacuum drying on the solid to obtain the polyamino acid nano particles. Wherein the temperature of the vacuum drying is preferably 20-40 ℃, and more preferably 25-35 ℃; the vacuum drying time is preferably 12-24 hours, and more preferably 18-24 hours. And (4) carrying out the post-treatment to obtain the polyamino acid nanoparticles.
With respect to step B): and dissolving the polyamino acid nano particles in halogenated acetic acid, and carrying out acidolysis in the presence of hydrogen bromide and acetic acid to remove carbobenzoxy to obtain the compound shown in the formula IV.
In the present invention, the halogenated acetic acid is preferably trifluoroacetic acid and/or dichloroacetic acid, and more preferably trifluoroacetic acid. The mass ratio of the volume of the halogenated acetic acid to the polyamino acid nanoparticles is preferably (8-12) mL to 1g, and more preferably 10mL to 1 g.
In the present invention, the source of the hydrogen bromide in acetic acid is not particularly limited, and may be a commercially available product. In the invention, in the acetic acid solution of the hydrogen bromide, the volume ratio of acetic acid to the hydrogen bromide is preferably 1: 0.5-5, and more preferably 1: 2.
In the invention, the acidolysis temperature is preferably 20-50 ℃, and more preferably 30-35 ℃; the acidolysis time is preferably 0.5-4 h, more preferably 1-2 h, and most preferably 1 h. Through the acidolysis reaction, the poly amino acid nanoparticles remove carbobenzoxy.
In the present invention, after the removal of benzyloxycarbonyl group by the acid hydrolysis reaction, it is preferable to further include: and (3) placing the obtained reaction solution into diethyl ether for suction filtration, dissolving the solid product obtained by suction filtration with deionized water, dialyzing, and freeze-drying to obtain the poly (L-lysine-co-gamma-glutamic acid) with the structure shown in the formula IV. Wherein, the dialysis is preferably carried out by adopting a dialysis bag with the molecular weight cutoff of 3500; the dialysis time is 4 days, and the dialysate is changed every 4 h. The freeze-drying method is not particularly limited in the invention, and the freeze-drying method known to those skilled in the art can be adopted; the temperature of the freeze-drying is preferably-20 ℃, and the time of the freeze-drying is preferably 72 h.
Wherein;
120≤x≤160，5≤y≤15，1≤a≤y，
or
120≤x≤160，y＝a＝0。
With respect to step C): under the action of a catalyst, the functionalized nano shell compound and the compound of the formula IV are mixed and reacted to obtain the cisplatin carrier with MMP-2 response shown in the formula I.
In the invention, the functionalized nano shell compound is a compound of formula II and/or a compound of formula III:
wherein, when preparing the final product y ≠ 0 (i.e., formula I-1 as described above), the compound of formula II is used to react with the compound of formula IV. When preparing the final product where y is 0 (i.e. formula i-2 as described above), a compound of formula III is used to react with a compound of formula IV.
According to the invention, the compounds of the formula II are preferably prepared by: the thiolated polyethylene glycol is reacted with a maleimide-based polypeptide (MI-PLGLAG-COOH) to produce a compound of formula II.
In the invention, the structure of the thiolated polyethylene glycol is shown as the following formula d, wherein the polymerization degree n is preferably 40-120, and the number average molecular weight of the corresponding polyethylene glycol is preferably 2000-5000. The source of the thiolated polyethylene glycol is not particularly limited, and the thiolated polyethylene glycol may be a commercially available product.
In the present invention, the structure of the maleimide-based polypeptide (MI-PLGLAG-COOH) is shown as the following formula e:
wherein PLGLAG is a polypeptide segment, specifically Pro-Leu-Gly-Leu-Ala-Gly. The maleimide-based polypeptide of the present invention is not particularly limited in its origin, and may be generally commercially available or prepared by a conventional method (e.g., solid-state synthesis) well known to those skilled in the art.
In the present invention, the reaction is preferably carried out in an organic solvent medium. The organic solvent is preferably one or more of N, N-dimethylformamide (i.e. DMF) and dichloromethane.
In the invention, the molar ratio of the maleimide-based polypeptide (MI-PLGLAG-COOH) to the thiolated polyethylene glycol is preferably (1.1-2.0) to 1, more preferably (1.3-1.6) to 1, and most preferably 1.5 to 1. The dosage ratio of the maleimide-based polypeptide (MI-PLGLAG-COOH) to the organic solvent is preferably 1g to (5-10) mL.
In the invention, the reaction temperature is preferably 15-40 ℃; the reaction time is preferably 5-7 h. Stirring is preferably accompanied during the reaction. After the above reaction, a solution containing the compound of formula II is formed, preferably also: the glacial ethyl ether is settled and dried to obtain the compound of the formula II.
According to the invention, the compounds of the formula III are preferably prepared by: mixing thiolated polyethylene glycol with maleimide-based polypeptide (MI-PLGLAG-NH)2) Reacting to generate the compound shown in the formula III.
In the present invention, the maleimide-based polypeptide (MI-PLGLAG-NH)2) The structure of (a) is shown as the following formula f:
the source of the maleimide-based polypeptide of the formula f is not particularly limited in the present invention, and it may be a commercially available product or prepared by a conventional method (e.g., solid-state synthesis) known to those skilled in the art.
Except for the above maleimide-based polypeptide raw materials, other contents such as the type of thiolated polyethylene glycol, the amount of thiolated polyethylene glycol used together with maleimide-based polypeptide, the above-mentioned operation process, reaction conditions, etc. are consistent when the compound of formula II is uniformly prepared, and are not described herein again.
The present invention, after obtaining a functionalized nanoshell compound (i.e., a compound of formula II and/or a compound of formula III), reacts it with a compound of formula IV. In the present invention, the reaction is preferably carried out in an organic solvent medium. In the present invention, the organic solvent is preferably N, N-dimethylformamide.
In the invention, the molar ratio of the functionalized nano shell compound (i.e. the compound of formula II and/or the compound of formula III) to the core of the compound of formula IV is preferably (1-20) to 1, more preferably (5-15) to 1, and most preferably 10: 1. The volume ratio of the total mass of the functionalized nano shell compound and the compound shown in the formula IV to the organic solvent is 1g to (8-12) mL, and the preferable volume ratio is 1g to 10 mL.
In the present invention, the reaction is carried out under the action of a catalyst. The catalyst is preferably 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine. The mass ratio of the catalyst to the compound shown in the formula IV is preferably (0.3-0.5) to 1. In the catalyst, the mass ratio of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride to 4-dimethylaminopyridine is preferably 1: 1-2. Preferably, the functionalized nano shell compound (i.e. the compound of formula II and/or the compound of formula III) is dissolved in the solvent, and the catalyst is added, mixed uniformly and dialyzed by the solvent; and adding a compound shown in the formula IV for reaction to form a compound shown in the formula I.
In the invention, the mixing temperature is preferably 10-30 ℃, and the mixing time is preferably 1-3 h. The reaction temperature is preferably 10-30 ℃; the reaction time is preferably 1-3 h. After the above reaction, it preferably further comprises: the resulting reaction solution was subjected to aqueous phase dialysis and freeze-drying. After the post-treatment, the cisplatin carrier with MMP-2 response shown in I is obtained:
the invention also provides a cis-platinum complex which is formed by matching cis-diamminedichloroplatinum with a carrier; wherein, the carrier is the cisplatin carrier with the MMP-2 response shown in the formula I in the technical scheme or the cisplatin carrier with the MMP-2 response shown in the formula I prepared by the preparation method in the technical scheme.
The structure of cis-diamminedichloroplatinum (namely cisplatin) is shown as the following formula g:
the cisplatin shown in the formula g is matched with the cisplatin carrier shown in the formula I, specifically, the cisplatin shown in the formula g reacts with carboxyl on the cisplatin carrier shown in the formula I, Cl of the cisplatin is removed, COO-of the compound shown in the formula I is connected, and the cisplatin carrier shown in the formula I is used for encapsulating the cisplatin to form a cross-linking structure due to the fact that the cisplatin contains two Cl, the cisplatin nanoparticle shown in the formula I serves as a shell, and the cisplatin is encapsulated in the nanoparticle.
The invention also provides a preparation method of the cisplatin complex in the technical scheme, which comprises the following steps: mixing the carrier and cis-diammine platinum dichloride for reaction to form a cis-platinum complex; wherein, the carrier is the compound shown in the formula I in the technical scheme.
In the invention, the reaction is preferably carried out in an aqueous medium, and specifically, the carrier compound shown in the formula I is dissolved in water, and then cis-diamminedichloroplatinum is added for reaction. In the invention, the mass ratio of glutamic acid units (i.e. units corresponding to the polymerization degree x-b) to cis-diamminedichloroplatinum in the carrier of the formula I is preferably (1-10) to 1, more preferably (4-8) to 1, and most preferably 6 to 1.
In the present invention, it is preferable to further adjust the pH after the addition and before the reaction; in the present invention, the pH is preferably adjusted to 8 to 10, more preferably 8.5 to 9.5, and most preferably 9. The kind of the pH adjustor used in the present invention for adjusting pH is not particularly limited, and may be a conventional alkaline adjustor well known to those skilled in the art. After the pH was adjusted, the reaction was carried out.
In the invention, the reaction temperature is preferably 30-45 ℃; the reaction time is preferably 2 to 5 days. In the present invention, after the above reaction, it is preferable to further perform: the resulting reaction solution was dialyzed and lyophilized. The method of dialysis and lyophilization is not particularly limited in the present invention, and may be performed in a conventional manner well known to those skilled in the art. Wherein the temperature of the freeze-drying is preferably-20 ℃, and the time of the freeze-drying is preferably 72 h. After the post-treatment, the cisplatin complex with MMP-2 response is obtained.
The cis-platinum coordination compound with MMP-2 response provided by the invention is formed by the coordination of cis-diamminedichloroplatinum and a compound with a structure shown in a formula I; the compound shown in the formula I is used as a carrier, polyethylene glycol is grafted on a side chain of polyamino acid, the carrier has good biocompatibility, degradability and solubility, and the carrier is grafted with a specific branched chain-thiol polyethylene glycol-maleimide polypeptide on the side chain of the polyamino acid, can be used as a shell of cisplatin, has certain shielding capability and MMP-2 responsiveness, is enriched at a tumor tissue part through enhancing an osmotic retention (EPR) effect, can prevent the toxicity of the cisplatin in other normal environments of a human body, can quickly respond and automatically remove the cisplatin under the stimulation of active oxygen over-expressed at a tumor prevention part, releases the cisplatin, and plays a role in a tumor part. Therefore, it can effectively avoid the sudden release and non-specific interaction in the blood circulation system after intravenous injection, thereby reducing the toxic and side effects and improving the therapeutic effect. Experimental results show that the cis-diammine-platinum dichloride complex has the characteristics of slow release capability and accelerated release under MMP-2 response.
For a further understanding of the invention, reference will now be made to the preferred embodiments of the invention by way of example, and it is to be understood that the description is intended to further illustrate features and advantages of the invention, and not to limit the scope of the claims.
Examples 1 to 3: preparation of polyethylene glycol-PLGLAG (i.e., a Compound of formula II) of varying molecular weights
Example 1
2.0g of mercaptopolyethylene glycol (molecular weight 5000) and 0.3g of maleimide-based polypeptide (MI-PLGLAG-COOH) were dissolved in 30mL of dichloromethane, reacted at 25 ℃ with stirring for 36 hours, precipitated twice with diethyl ether, and dried to obtain the compound of formula II.
Example 2
1.6g of mercaptopolyethylene glycol (molecular weight 4000) and 0.3g of maleimide-based polypeptide (MI-PLGLAG-COOH) were dissolved in 30mL of dichloromethane, reacted at 25 ℃ with stirring for 36h, precipitated twice with diethyl ether, and dried to obtain the compound of formula II.
Example 3
0.8g of mercaptopolyethylene glycol (molecular weight 2000) and 0.3g of maleimide-based polypeptide (MI-PLGLAG-COOH) were dissolved in 30mL of dichloromethane, reacted at 25 ℃ with stirring for 36h, precipitated twice with diethyl ether, and dried to obtain the compound of formula II.
Example 4: preparation of N (epsilon) -benzyloxycarbonyl-L-lysine-N-cyclic lactam
Mixing 1g N-benzyloxycarbonyl-L-lysine with 0.6g of bis (trichloromethyl) carbonate at 25 ℃, adding tetrahydrofuran, heating to 50 ℃ for reaction for 2h, after the reaction is finished, settling the reaction mixture in excessive petroleum ether, separating, washing, recrystallizing and drying to obtain N (epsilon) -benzyloxycarbonyl-L-lysine-N-cyclic lactam, wherein a nuclear magnetic spectrum is shown in figure 1, and figure 1 is a nuclear magnetic spectrum of the N (epsilon) -benzyloxycarbonyl-L-lysine-N-cyclic lactam prepared in example 4.
Example 5: preparation of gamma-benzylglutamic acid-N-cyclic anhydride
Mixing 1g of gamma-benzyl glutamic acid and 0.6g of bis (trichloromethyl) carbonate at 25 ℃, adding tetrahydrofuran, heating to 50 ℃ for reaction for 2 hours, settling the reaction mixture in excessive petroleum ether after the reaction is finished, separating, washing, recrystallizing and drying to obtain the gamma-benzyl glutamic acid-N-cyclic internal anhydride, wherein the nuclear magnetic spectrum refers to figure 2, and figure 2 is the nuclear magnetic spectrum of the gamma-benzyl glutamic acid-N-cyclic internal anhydride prepared in example 5.
Examples 6 to 14: preparation of polyamino acid nanoparticles with different contents
Example 6
27.22g of γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 2.64g of N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, reacted at 25 ℃ under nitrogen for 3 days with stirring, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, wherein 120 γ -benzylglutamic acids and 10N (e) -benzyloxycarbonyl-L-lysine (i.e., x 120, y 10) were polymerized per N-hexylamine on average.
Example 7
27.22g of γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 1.32g of N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, reacted at 25 ℃ under nitrogen for 3 days with stirring, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average, 120 γ -benzylglutamic acids and 5N (e) -benzyloxycarbonyl-L-lysine (i.e., x 120, y 5) were polymerized per N-hexylamine.
Example 8
27.22g of γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 3.96g of N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, reacted at 25 ℃ under nitrogen for 3 days with stirring, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average, 120 γ -benzylglutamic acids and 15N (e) -benzyloxycarbonyl-L-lysine (i.e., x 120, y 15) were polymerized per N-hexylamine.
Example 9
34.03g of the γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 2.64g of the N (. epsilon. -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, and stirred under nitrogen atmosphere at 25 ℃ for 3 days, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average, 150 γ -benzylglutamic acids and 10N (. epsilon. -benzyloxycarbonyl-L-lysine (i.e., x. 150, y. 10) were polymerized per N-hexylamine.
Example 10
34.03g of the γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 1.32g of the N (. epsilon. -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed homogeneously, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, stirred under nitrogen atmosphere at 25 ℃ for 3 days, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average, 150 γ -benzylglutamic acids and 5N (. epsilon. -benzyloxycarbonyl-L-lysine (i.e., x. 150, y. 5) were polymerized per N-hexylamine.
Example 11
34.03g of the γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 3.96g of the N (. epsilon. -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed homogeneously, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, stirred under nitrogen atmosphere at 25 ℃ for 3 days, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average, 150 γ -benzylglutamic acids and 15N (. epsilon. -benzyloxycarbonyl-L-lysine (i.e., x. 150, y. 15) were polymerized per N-hexylamine.
Example 12
36.30g of γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 2.64g of N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, reacted at 25 ℃ under nitrogen for 3 days with stirring, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average, 160 γ -benzylglutamic acids and 10N (e) -benzyloxycarbonyl-L-lysine (i.e., x 160, y 10) were polymerized per N-hexylamine.
Example 13
36.30g of γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 1.32g of N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, reacted at 25 ℃ under nitrogen for 3 days with stirring, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average 160 γ -benzylglutamic acids and 5N (e) -benzyloxycarbonyl-L-lysine (i.e., x 160, y 5) were polymerized per N-hexylamine.
Example 14
36.30g of γ -benzylglutamic acid-N-cyclic anhydride prepared in example 5 and 3.96g of N (e) -benzyloxycarbonyl-L-lysine-N-cyclic anhydride prepared in example 4 were mixed uniformly, added to a solution of N, N-dimethylformamide containing 0.087g of N-hexylamine, reacted at 25 ℃ under nitrogen for 3 days with stirring, the resulting solution was poured into 100mL of anhydrous ether, and the solid was collected by suction filtration and dried under vacuum to obtain polyamino acid nanoparticles, on average 160 γ -benzylglutamic acids and 15N (e) -benzyloxycarbonyl-L-lysine (i.e., x 160, y 15) were polymerized per N-hexylamine.
Examples 15 to 23: the polyamino acid nanoparticles prepared in examples 6 to 14 were subjected to N-benzyloxycarbonyl and benzyl removal to prepare compounds of formula IV
1g of polyamino acid nanoparticles prepared in examples 6 to 14 [ i.e., poly (N (epsilon) -benzyloxycarbonyl-L-lysine-co-gamma-benzylglutamic acid) ] are respectively weighed and dissolved in 10mL of trifluoroacetic acid, 3mL of acetic acid solution of hydrogen bromide (concentration of 33 wt%) is added, and the mixture is stirred and reacted for 1 hour at room temperature; pouring the reaction solution into 100mL of diethyl ether, performing suction filtration, dissolving the obtained solid with water, dialyzing the solution in deionized water for 3d by using a dialysis bag with the molecular weight cutoff of 3500, changing the dialyzate every 4h, freeze-drying the obtained solution to obtain poly (L-lysine-co-gamma-glutamic acid) shown in the formula IV, wherein the nuclear magnetic spectrum is shown in figure 3, the figure 3 is the nuclear magnetic spectrum of the poly (L-lysine-co-gamma-glutamic acid) shown in the formula IV prepared in example 13, and the number average molecular weight of the poly (L-lysine-co-gamma-glutamic acid) prepared in example 13 is 23000 through nuclear magnetic calculation.
Examples 24 to 50: preparation of cisplatin vectors of formula I having MMP-2 response
Examples 24 to 32
Weighing a certain amount of the compound of the formula II prepared in example 1 (the molar weight of lysine in the compound of the formula IV prepared in examples 15-23 is equal, namely the molar weight of the structural unit corresponding to y is equal), dissolving the compound of the formula II in 5mL of DMF, adding 0.4g of catalyst (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine in a mass ratio of 1: 1.5), stirring and reacting at 25 ℃ for 2h, dialyzing with DMF, adding 1g of the compound of the formula IV prepared in examples 15-23, stirring and reacting at 25 ℃ for 2h, dialyzing with aqueous phase, and freeze-drying to obtain the cisplatin-supported nanoparticles shown in formula I with MMP-2 response.
The nuclear magnetic spectrum of the product obtained in example 24 is shown in fig. 4, and fig. 4 is the nuclear magnetic spectrum of the cisplatin carrier nanoparticle shown in formula I prepared in example 25, wherein x is 150, y is 10, and a is 7.
Examples 33 to 41
Weighing a certain amount of the compound of the formula II prepared in example 2 (the molar weight of lysine in the compound of the formula IV prepared in examples 15-23 is equal, namely the molar weight of the structural unit corresponding to y is equal), dissolving the compound of the formula II in 5mL of DMF, adding 0.4g of catalyst (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine in a mass ratio of 1: 1.5), stirring and reacting at 25 ℃ for 2h, dialyzing with DMF, adding 1g of the compound of the formula IV prepared in examples 15-23, stirring and reacting at 25 ℃ for 2h, dialyzing with aqueous phase, and freeze-drying to obtain the cisplatin-supported nanoparticles shown in the formula I with MMP-2 response.
Examples 42 to 50
Weighing a certain amount of the compound of the formula II prepared in example 3 (the molar weight of lysine in the compound of the formula IV prepared in examples 15-23 is equal, namely the molar weight of the structural unit corresponding to y is equal), dissolving the compound of the formula II in 5mL of DMF, adding 0.4g of catalyst (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine in a mass ratio of 1: 1.5), stirring and reacting at 25 ℃ for 2h, dialyzing with DMF, adding 1g of the compound of the formula IV prepared in examples 15-23, stirring and reacting at 25 ℃ for 2h, dialyzing with aqueous phase, and freeze-drying to obtain the cisplatin-supported nanoparticles shown in the formula I with MMP-2 response.
Examples 51 to 77: preparation of cisplatin complexes with MMP-2 response
Respectively weighing 100mg of cisplatin carrier nanoparticles shown in formula I prepared in examples 24-50, adding cisplatin according to the mass ratio of glutamic acid units to cisplatin in the carrier of 4: 1, adjusting the pH value of the system to 9, carrying out constant-temperature oscillation reaction at 37 ℃ for 3d, dialyzing, and freeze-drying to obtain the polyamino acid material (namely the cisplatin complex) carrying dichlorodiamineplatinum with MMP-2 response.
Example 78: cisplatin complexes with MMP-2 response release under different conditions
5mg of the cisplatin complex prepared in example 51 was weighed at 37 ℃ and dissolved in 5mL of HEPES buffer solution containing MMP-2 (MMP-2 content: 0.2. mu.g/mL) for 1 hour, and then transferred to a dialysis bag (molecular weight cut-off of the dialysis bag is 3500), two samples were taken, dialyzed against buffer solutions (38mL) having pH 7.4 and pH 5.5, respectively, and 2mL were sampled at 6h, 12h, 24h, 48h, 72h and 96h, and the same amount of buffer solution was added to form samples to be measured at different time nodes. And the MMP-2-free system was used as a control.
The samples to be tested and the control samples are quantitatively analyzed by using inductively coupled plasma mass spectrometry, the change relation of the accumulated release percentage of the cis-platinum along with the increase of time is obtained, the release result is shown in figure 5, figure 5 is a release curve graph of the cis-platinum complex prepared by the invention under different conditions, and as can be seen from figure 5, the cis-platinum complex prepared by the invention has the slow release capacity and the property of accelerating the release under MMP-2 response. The above experiment was carried out on the cisplatin complexes prepared in other examples, and the results also show that the cisplatin complexes prepared in the present invention have sustained release ability and accelerated release property under MMP-2 response.
Example 79: MTT experiment verification of antitumor effect
Using the human ovarian cancer cell line SKOV3 as an example, cells were uniformly plated in a 96-well plate at about 7000 cells per well, and the cisplatin complex prepared in example 51 was added so that the final concentration became a gradient (0.313. mu.g/mL, 0.625. mu.g/mL, 1.25. mu.g/mL, 2.5. mu.g/mL, 5.0. mu.g/mL, 10.0. mu.g/mL, 20.0. mu.g/mL), and the cell activity was tested after two days of incubation. The results are shown in FIG. 6, and FIG. 6 is a graph showing the effect of MTT antitumor assay in example 79. As can be seen from FIG. 6, the cell activity showed a decreasing trend with the increase in the concentration of the cisplatin complex of the present invention, indicating that the cisplatin complex of the present invention has a good MMP-2 responsiveness and a good antitumor effect. The above experiments were carried out on the cisplatin complexes prepared in other examples, and the results also show that the cisplatin complexes prepared in the invention have good MMP-2 responsiveness and anti-tumor effects.
The experimental effects show that the cisplatin carrier shown in the formula I has good slow release capacity and the characteristic of accelerating release under MMP-2 response, and shows good MMP-2 responsiveness and anti-tumor effect.
The above description of the embodiments is only intended to facilitate the understanding of the method of the invention and its core idea. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A cisplatin carrier having MMP-2 response, characterized by having the structure shown in formula I:
wherein:
40≤n≤120；
120≤x≤160，b＝0，5≤y≤15，1≤a≤y，
or
120≤x≤160，1≤b≤x，y＝a＝0；
PLGLAG is polypeptide segment, specifically Pro-Leu-Gly-Leu-Ala-Gly.
2. The cisplatin carrier as described in claim 1,
140≤x≤155，b＝0，8≤y≤12，1≤a≤y，
or
140≤x≤155，1≤b≤x，y＝a＝0。
3. A method for preparing a cisplatin carrier as an MMP-2 response as described in any of claims 1-2, comprising the steps of:
A) mixing N-hexylamine, N-benzyloxycarbonyl-L-lysine-N-cyclic carboxylic anhydride and gamma benzyl glutamic acid-N-cyclic carboxylic anhydride for reaction to obtain polyamino acid nanoparticles;
B) dissolving the polyamino acid nanoparticles in halogenated acetic acid, and carrying out acidolysis in the presence of hydrogen bromide and acetic acid to remove carbobenzoxy groups, thereby obtaining a compound shown in a formula IV;
C) under the action of a catalyst, a functionalized nano shell compound and a compound shown in a formula IV are mixed and react to obtain a cisplatin carrier with MMP-2 response shown in the formula I;
the functionalized nano shell compound is a compound of a formula II or a compound of a formula III;
wherein:
40≤n≤120；
120≤x≤160，5≤y≤15，1≤a≤y，
or
120≤x≤160，y＝a＝0。
4. The method according to claim 3, wherein the reaction temperature in step A) is 15 to 50 ℃ and the reaction time is 2 to 7 days.
5. The preparation method according to claim 3, wherein in the step C), the reaction temperature is 10-30 ℃ and the reaction time is 1-3 h;
the catalyst is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 4-dimethylaminopyridine.
6. A cisplatin complex, which is characterized in that cis-diamminedichloroplatinum is matched with a carrier to form the cisplatin complex;
the carrier is the cisplatin carrier with MMP-2 response described in any one of claims 1-2 or the cisplatin carrier with MMP-2 response prepared by the preparation method described in any one of claims 3-5.
7. A method for producing a cisplatin complex as described in claim 6, comprising:
mixing the carrier and cis-diammine platinum dichloride for reaction to form a cis-platinum complex;
the vector is a cisplatin vector having MMP-2 response as defined in any of claims 1-2.
8. The preparation method according to claim 7, wherein the reaction is carried out at a pH of 8 to 10;
the reaction temperature is 30-45 ℃, and the reaction time is 2-5 days.
9. The method according to claim 7, wherein the molar ratio of glutamic acid units to cis-diamminedichloroplatinum in the carrier is (1-10) to 1.
10. The method according to claim 7, wherein the reaction is carried out in water as a solvent.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009045579A2 (en) * | 2007-06-14 | 2009-04-09 | The Regents Of The University Of California | Multimodal imaging probes for in vivo targeted and non-targeted imaging and therapeutics |
| CN102895669A (en) * | 2012-11-01 | 2013-01-30 | 中国科学院长春应用化学研究所 | Cis-platinum complex and preparation method thereof |
| CN105001426A (en) * | 2015-08-18 | 2015-10-28 | 中国科学院长春应用化学研究所 | Tumor targeting polyamino acid graft copolymer and preparation method thereof |
| CN106420611A (en) * | 2016-11-11 | 2017-02-22 | 天津城建大学 | PH-sensitive chitosan drug-carrying micelle with targeting and fluorescent characteristics and preparation method thereof |
| CN109438715A (en) * | 2018-12-24 | 2019-03-08 | 中国科学院长春应用化学研究所 | A kind of cis-dichlorodiamine platinum complex, preparation method and application with ROS response |
| CN109481695A (en) * | 2018-12-24 | 2019-03-19 | 中国科学院长春应用化学研究所 | A kind of cis-dichlorodiamine platinum complex, preparation method and application with pH response |
| CN111744020A (en) * | 2019-03-27 | 2020-10-09 | 中国科学院宁波材料技术与工程研究所 | Active targeting response type polypeptide drug, preparation method and application thereof |
-
2020
- 2020-11-19 CN CN202011305361.9A patent/CN112480419B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009045579A2 (en) * | 2007-06-14 | 2009-04-09 | The Regents Of The University Of California | Multimodal imaging probes for in vivo targeted and non-targeted imaging and therapeutics |
| CN102895669A (en) * | 2012-11-01 | 2013-01-30 | 中国科学院长春应用化学研究所 | Cis-platinum complex and preparation method thereof |
| CN105001426A (en) * | 2015-08-18 | 2015-10-28 | 中国科学院长春应用化学研究所 | Tumor targeting polyamino acid graft copolymer and preparation method thereof |
| CN106420611A (en) * | 2016-11-11 | 2017-02-22 | 天津城建大学 | PH-sensitive chitosan drug-carrying micelle with targeting and fluorescent characteristics and preparation method thereof |
| CN109438715A (en) * | 2018-12-24 | 2019-03-08 | 中国科学院长春应用化学研究所 | A kind of cis-dichlorodiamine platinum complex, preparation method and application with ROS response |
| CN109481695A (en) * | 2018-12-24 | 2019-03-19 | 中国科学院长春应用化学研究所 | A kind of cis-dichlorodiamine platinum complex, preparation method and application with pH response |
| CN111744020A (en) * | 2019-03-27 | 2020-10-09 | 中国科学院宁波材料技术与工程研究所 | Active targeting response type polypeptide drug, preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| "MMP-2酶刺激相应性纳米给药系统的构建及其体内外评价";皇甫铭一;《CNKI硕士论文全文数据库医药科技辑》;20180705(第07期);第E079-56页 * |
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