CN108409756A - Multi-functional prodrug of a kind of heterodimer based on camptothecin and its preparation method and application - Google Patents
Multi-functional prodrug of a kind of heterodimer based on camptothecin and its preparation method and application Download PDFInfo
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- CN108409756A CN108409756A CN201810188970.7A CN201810188970A CN108409756A CN 108409756 A CN108409756 A CN 108409756A CN 201810188970 A CN201810188970 A CN 201810188970A CN 108409756 A CN108409756 A CN 108409756A
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- camptothecin
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
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- A—HUMAN NECESSITIES
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
- A61K49/0036—Porphyrins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/0474—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group
- A61K51/0485—Porphyrins, texaphyrins wherein the nitrogen atoms forming the central ring system complex the radioactive metal
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K9/00—Medicinal preparations characterised by special physical form
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- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The present invention provides a kind of multi-functional prodrug of the heterodimer based on camptothecine, it is loaded with camptothecine simultaneously and 2 (1 hexyloxyehtyls) 2 go the dilute Ji Jiaotuo of second to delay chlorophyllin a (HPPH), it is the camptothecine HPPH prodrugs that camptothecin is formed with HPPH by linking group, shown in structural formula such as formula (I)；Wherein, R is linking group, is SS, CH2CH2Or S (CH3)C(CH3) one kind in S；R1It is camptothecin group.The prodrug of the present invention easily can be wrapped to form nano particle by amphipathy macromolecule, and nano particle carrier medicine carrying efficiency height, the drugloading rate of formation are big, and under glutathione existence condition, unmodified camptothecine and HPPH can be gone out with quick release；Nano particle can be absorbed effectively by tumour cell and kill tumour cell simultaneously.The present invention also provides the preparation method of the prodrug, preparation and its applications in preparing cancer treatment drugs.
Description
Technical field
It is specifically a kind of the present invention relates to biological medicine technology, nanometer medicine and drug controlled release technical field
The multi-functional prodrug of the heterodimer based on camptothecin of deoxidization, degradation drug release, and its preparation method and application.
Background technology
Cancer is the world's second largest cause of death.According to the statistics of the World Health Organization, cancer causes 8,800,000 people dead to root within 2015
It dies.People death of the whole world close to 1/6th is due to cancer, wherein about 70% cancer mortality is happened at low income and medium
Income country.Currently, chemotherapy is one of primary treatments of cancer.However, most of chemotherapeutics, such as camptothecine
(camptothecin), taxol (paclitaxel) etc., solubility is limited in water, and has adverse side effect.Therefore,
Modification and the effective drug delivery system of design to chemotherapeutics are always the research focus in drug controlled release field.It receives
Rice drug has been widely studied for treatment of cancer.The use of nano particle can not only improve the water dispersible of these drugs,
But also distribution in their pharmacokinetics and organism can be enhanced, improve therapeutic effect and reduce side effect.
Polymer micelle (polymeric micelles) nano-particle is most important in Nano medication treating cancer field
One of pharmaceutical carrier.Most polymer micelle includes two parts, when the hydrophobic core for drug loading, second is that for changing
The polyethylene glycol (PEG) of kind colloidal stability.Currently, several micellar preparations, for example, Genexol-PM, NK012 and NK105 into
Enter clinical experimental stage, but not yet gets the Green Light in the U.S..The part challenge that these micellar preparations face is low drugloading rate, too early
Drug release, internal drug metabolism is soon and accumulation is few etc..In order to solve these limitations, in addition to developing new drug carrier, also
Small-molecule drug can be modified.Compared with pharmaceutical carrier optimizes, the modification of small-molecule drug is relatively easy, can promote
Drug discovery process.
Camptothecine (camptothecin, CPT) is the alkaloid extracted from camplotheca acuminata earliest, it is that DNA topologys are different
Structure enzyme I (TOPO I) inhibitor has significant antitumor activity in preclinical study.But due to low solubility and it is serious not
Good side effect fails in clinical test.Currently, camptothecin analogues Irinotecan (Irinotecan) and topotecan
(Topotecan) ratified for treating colon cancer by food and drug administration (FDA).Irinotecan in water can
Hydrolysis 7-Ethyl-10-hydroxycamptothecin (SN-38), the latter have antitumor activity of 1000 times than Irinotecan this height.
But Irinotecan can only hydrolyze a small amount of 7-Ethyl-10-hydroxycamptothecin in human body, therefore its lethality to cancer cell
It is apparent impaired.HPPH is a kind of photosensitizer, and full name is that 2- (1- hexyloxyehtyls) -2- goes the dilute Ji-Jiao Tuo of second to delay chlorophyllin-a (2-
[1-hexyloxyethyl]-2-devinyl pyropheophorbide-a).HPPH can consume molecular oxygen after laser irradiation
To generate singlet oxygen to kill cancer cell.CPT, can stable topology isomerase I-DNA as a kind of topoisomerase I inhibitor
Compound causes DNA damage, to kill cancer cell.In addition, CPT is a kind of hypoxia-inducible factor-1 alpha (HIF-1 α) inhibition
Agent, can reduce cancer cell low-oxygen environment survival ability.Therefore, CPT is expected to the cellulotoxic effect of enhancing HPPH inductions.
In addition, the chelating agent that HPPH itself can be marked as fluorescent dye and Cu-64, can pass through fluorescence and quantitative PET imagings prison
Survey drug.Therefore, we are a kind of in the invention while being loaded with camptothecin and the compound of HPPH, as glutathione
(GSH) the multi-functional prodrug of sensibility heterodimer not only has extra high load efficiency and high load capability, can also be
Directly PET drugs imaging in vivo, but also can realize collaboration treatment of cancer.It is worth noting that, the characteristic of PET drugs imaging
Make it possible to the multi-functional prodrug of accurate measurements heterodimer and determines its internal pharmacokinetics and bio distribution.In addition, working as
When the heterodimer is connected by disulfide bond, disulfide bond is easy to be cracked by the GSH of cancer cell middle and high concentration, then passes through cascade
Reaction discharges CPT and HPPH from the multi-functional prodrug of heterodimer.
Currently, the multi-functional prodrug of this heterodimer there is no to be reported both at home and abroad.
Invention content
The primary and foremost purpose of the present invention is to provide a kind of prodrug that camptothecine and HPPH can be discharged in tumor cells selectivity, tool
There is significant inhibition of cancer cell effect, while adverse side effect is low.
It is a further object of the present invention to provide the methods for preparing the prodrug, and building-up process is simple and practicable, and drugloading rate is high.
Another object of the present invention is to provide application of the prodrug in preparing cancer treatment drugs.
The above-mentioned purpose of the present invention is achieved through the following technical solutions：
First, a kind of multi-functional prodrug of the heterodimer based on camptothecin is provided, it be loaded with simultaneously camptothecin and
HPPH is the camptothecin-HPPH prodrugs that camptothecine or derivatives thereof is formed with HPPH by linking group, structure such as formula
(I) shown in：
Wherein,
R isOrIn one kind；R1It is camptothecin group.
In the solution of the present invention, described camptothecine or derivatives thereof can be selected from camptothecine or 7- ethyl -10- hydroxyls are liked
Set any one in alkali.
In preferred embodiments of the present invention, the multi-functional prodrug is that camptothecine passes through the linking group R shapes with HPPH
At prodrug, i.e., the R in the described formula (I) isR1It is camplotheca acuminata base groups；Shown in its structure such as following formula (II)
The present invention also provides a kind of methods preparing the camptothecin-HPPH prodrugs, are with camptothecine or derivatives thereof
It by being reacted to the raw material with corresponding group, is made among camptothecine or derivatives thereof prodrug in organic solvent for raw material
Camptothecin-HPPH the prodrugs are prepared with HPPH reactions again in body, the last intermediate.
The method of the present invention for preparing camptothecin-HPPH prodrugs, specifically includes following steps：
1) triphosgene activation camptothecine or derivatives thereof lactonic ring in organic solvent, is used in the presence of acylation catalyst
On hydroxyl, add bis- thiodiethanols of excessive 2,2'- or 1,6- hexylene glycols reaction or 2,2'- (propane -2,2- bis-
Base is bis- (sulphur)) diethanol, obtain intermediate 1；
2) intermediate 1 and HPPH that step 1) obtains are mixed in organic solvent, is reacted by coupling reagent and generates ester
Key obtains camptothecin-HPPH prodrugs in part of the present invention.
In the currently preferred preparation method, the organic solvent described in step 1) is selected from dichloromethane, chloroform, tetrahydrochysene
Any one in furans, 1,4- dioxane or dimethylformamide.
In the currently preferred preparation method, the acylation catalyst described in step 1) is selected from N- ethyls-N '-[(3- bis-
Methylamino) propyl] two Asia of carbodiimide hydrochloride (EDC), 4- (dimethylamino) pyridine (DMAP) or N, N'- dicyclohexyl carbon
Any one of amine (DCC).
In the currently preferred preparation method, the organic solvent described in step 2) is selected from dichloromethane, chloroform, tetrahydrochysene
Any one in furans, 1,4- dioxane or dimethylformamide.
In the currently preferred preparation method, the coupling reagent described in step 2) is selected from 1- ethyls-(3- dimethylaminos
Base propyl) phosphinylidyne diimmonium salt hydrochlorate (EDC.HCL), N, N- dicyclohexyl carbonic acid diimines (DCC) or 1H- benzotriazole -1-
Any one in base oxygen tripyrrole alkyl hexafluorophosphate (PyBOP).
In a kind of currently preferred embodiment, the method synthetic route for preparing prodrug shown in formula (II) is as follows：
Specifically comprise the following steps：
In the presence of DMAP, in methylene chloride, the hydroxyl of camptothecine is activated using triphosgene, then with excessive 2,
Bis- thiodiethanols of 2'- are reacted, and intermediate CPT-ss-OH is prepared；Again by CPT-ss-OH and HPPH together in dichloromethane
Middle mixing, and EDC and DMAP stirrings are added, prodrug shown in formula (II) is prepared：CPT-ss-HPPH.
In currently preferred another embodiment, it is prepared for prodrug shown in following formula (III)：
Its synthetic route is as follows：
Specifically comprise the following steps：
In the presence of DMAP, in methylene chloride, the hydroxyl of camptothecine is activated using triphosgene, then with excessive 1,
6- hexylene glycols react, and intermediate CPT-cc-OH is prepared；CPT-cc-OH and HPPH are mixed in methylene chloride together again,
And EDC and DMAP stirrings are added, prodrug shown in formula (II) is prepared：CPT-cc-HPPH.
The prodrug of the present invention can directly be used alone as cancer treatment drugs, can also further be made in other pharmacology
Acceptable dosage form is used for treatment of cancer.Therefore, the present invention also provides the camptothecin-HPPH prodrugs can pharmaceutically connect
The preparation received.
In preferred embodiments of the present invention, the preparation is nanoparticle formulations, is excipient outside the nano particle
Before the camptothecine-HPPH that package originally has invention described inside agent and/or pharmaceutical carrier, the excipients and/or pharmaceutical carrier
Medicine.
The excipients and pharmaceutical carrier include but not limited to twain series emulsifier (such as Tween 80, polysorbas20),
Any one in Emulsifier EL-60 (Cremophor EL), liposome (liposome) or amphipathy macromolecule or two
Kind or more mixture.Preferred amphiphilic macromolecule of the present invention.
The amphipathy macromolecule is selected from polyethylene-b-polylactic acid (polyethylene glycol-b-
Polylactide, PEG-b-PLA), polyethylene glycol-b- polycaprolactones (polyethylene glycol-b-
Polycaprolactone, PEG-b-PCL) or the poly- phosphides of polyethylene glycol-b- (polyethylene glycol-b-
Polyphosphoester, PEG-b-PPE) in any one.
The present invention also provides application of the camptothecin-HPPH prodrugs in preparing the cancer treatment drugs.
In currently preferred application scheme, the camptothecin-HPPH prodrugs are carried out using amphipathy macromolecule
Nano particle is prepared in physically encapsulation.
In a kind of preferred embodiment of the present invention, using amphipathy macromolecule as pharmaceutical carrier to the camplotheca acuminata
Bases-HPPH prodrugs carry out physically encapsulation；Specifically by camptothecin-HPPH prodrugs of the present invention and amphipathy macromolecule
Be dissolved in organic solvent (such as tetrahydrofuran) together with certain proportion, then instill stirring or ultrasound aqueous solution in, so
After vapor away organic solvent, be made nano particle.The amphipathy macromolecule is selected from PEG-b-PLA, PEG-b-PCL or PEG-
Any one in b-PPE.
In the present invention, camptothecin and HPPH are joined together to form prodrug by connecting key, and especially the two passes through
When disulfide bond links together, camptothecin and HPPH can be selectively discharged in tumour cell.Currently, there is no both at home and abroad
This prodrug is reported.Prodrug and its nanoparticle formulations of the present invention have the following advantages that：
(1) camptothecin and HPPH have synergistic effect；
(2) camptothecin is modified by sulphation, before connecting key fracture, to the small toxicity of cell；
(3) when camptothecin is connected with HPPH by disulfide bond, the prodrug is to intracellular reducing substances, such as gluathione
Peptide is sensitive, and camptothecin and HPPH can be discharged rapidly after contacting glutathione；
(4) in the prior art, camptothecin and HPPH are not easy to be wrapped up by amphipathy macromolecule, and the drug wrapped up is easy
It is released out of particle, and the prodrug in the present invention, it is more hydrophobic due to molecular weight bigger, cause them from releasing in particle
It puts speed to substantially reduce, advantageously reduces the toxic side effect of drug, improve curative effect；
(5) prodrug that the present invention synthesizes can be extremely efficiently wrapped, and the drugloading rate of nano particle is also very high；
(6) synthesis of prodrug is simple, and the preparation of nano particle is easy, and is conducive to clinical conversion.
Description of the drawings
Fig. 1 is the ESI-MS spectrograms of CPT-ss-HPPH prepared by embodiment 1.
Fig. 2 is CPT-ss-HPPH's prepared by embodiment 11H NMR(300MHz,CDCl3) spectrogram.
Fig. 3 is the ESI-MS spectrograms of CPT-cc-HPPH prepared by embodiment 2.
Fig. 4 is CPT-cc-HPPH's prepared by embodiment 21H NMR (300MHz, CDCl3) spectrogram.
Fig. 5 is the ultraviolet-visible spectrogram of CPT, HPPH and the CPT-ss-HPPH of the preparation of embodiment 1.
Fig. 6 is to be monitored in the experiment that different pharmaceutical generates ROS abilities using anthracene -9,10- dipropionic acids (ADPA) in embodiment 3
With 671 nanometer lasers respectively according to the UV-Vis spectrograms of the different pharmaceutical obtained after 1,2,3,4,5,6 and 7 minute.Wherein, (a)
For the UV-Vis spectrograms of HPPH upon laser irradiation；(b) it is the UV-Vis spectrograms of CPT-ss-HPPH upon laser irradiation；
(c) it is absorbance quantitative analysis to HPPH and CPT-ss-HPPH at 410 nanometers；(d) it is to exist to HPPH and CPT-ss-HPPH
662 nanometers of absorbance quantitative analysis；(e) be HPPH and CPT-ss-HPPH degradation analysis.
Fig. 7 is the TEM image of CPT-ss-HPPH NPs prepared by embodiment 4.
Fig. 8 be embodiment 5 described in vitro drug release experiment in CPT NPs, HPPH NPs, CPT-cc-HPPH NPs and
Drug release rates of the CPT-ss-HPPH in PBS changes over time figure.
Fig. 9 is vitro cytotoxicity measuring result curve figure described in embodiment 6.
Figure 10 is the hydrated diameter distribution map of the CPT-ss-HPPH NP in embodiment 4.
Figure 11 is the zeta potential diagrams of the CPT-ss-HPPH NP in embodiment 4.
Figure 12 be in embodiment 7 the HCT116 tumour cells that arrive of fluorescence microscopy to HPPH NPs, CPT-cc-HPPH
The cellular uptake of NPs and CPT-ss-HPPH NPs.Engineer's scale is 30 microns.
Figure 13 is Positron emission computed tomography (PET) imaging of HCT116 tumor-bearing mices described in embodiment 8.Its
Middle white circle is knub position.
Figure 14 be embodiment 8 described in by PET to drug the enrichment of tumour quantitative analysis (n=3).
Figure 15 is to count determining drug distribution (n=3) by the γ to isolated organ described in embodiment 8.
Figure 16 is interior therapeutic experiment gained tumor growth curve figure described in embodiment 9；Wherein abscissa the 0th and the 4th day
Locate arrow and represent intravenous injection, arrow represents laser irradiation at the 2nd and the 6th day.
Figure 17 is the survival rate of mouse after interior therapeutic experiment gained ejection preparation described in embodiment 9.
Specific implementation mode
The present invention is specifically described below by embodiment, the present embodiment is served only for making further the present invention
It is bright, it should not be understood as limiting the scope of the invention, those skilled in the art makes according to the content of foregoing invention
Some nonessential modifications and adaptations, belong to the scope of the present invention.
Embodiment 1：The preparation of CPT-ss-HPPH
DMAP (1.15mg, 9.42 μm of ol), HPPH (30mg, 4.71 μm of ol) and CPT- are mixed in 20mL dichloromethane
Ss-OH (51.0mg, 94.3 μm of ol).It is molten with the dichloromethane that 5mL EDC HCl (18.1mg, 94.3 μm of ol) are added dropwise under stiring
Liquid.After being stirred at room temperature 24 hours, confirm that HPPH reacts completely by thin-layer chromatography (TLC).It is concentrated using rotary evaporator
Mixture solution then uses gradient ethyl acetate and hexane to pass through flash chromatography (Teledyne ISCO as eluent
CombiFlash it) purifies.Yield 39.9mg (74%).ESI-MS m/z(M+)calcd 1146.46,found 1147.24(M+
H+).1H NMR(300MHz,CDCl3) δ 9.80 (d, J=8.9Hz, 1H), 9.52 (s, 1H), 8.44 (d, J=5.7Hz, 1H),
8.00 (t, J=8.7Hz, 1H), 7.69-7.51 (m, 2H), 7.42-7.28 (m, 2H), 7.21 (d, J=7.5Hz, 1H), 5.91
(p, J=6.5Hz, 1H), 5.68 (d, J=17.2Hz, 1H), 5.38-5.3 (m, 1H), 5.09 (qd, J=20.0,3.0Hz,
2H), 4.81 (d, J=6.0Hz, 1H), 4.69 (dd, J=17.3,1.2Hz, 1H), 4.44-4.38 (m, 1H), 4.40-4.28
(m, 2H), 4.26-4.17 (m, 3H), 3.73 (q, J=7.4Hz, 2H), 3.67 (s, 3H), 3.65-3.55 (m, 1H), 3.36 (d,
J=2.7Hz, 3H), 3.28 (s, 3H), 2.88 (t, J=6.6Hz, 2H), 2.77 (t, J=6.6Hz, 2H), 2.58-2.18 (m,
4H), 2.13 (dd, J=9.2,6.7Hz, 4H), 1.80-1.75 (m, 3H), 1.75-1.68 (m, 3H), 1.59 (s, 6H), 1.22-
1.19 (m, 2H), 0.99 (t, J=7.4Hz, 3H), 0.77 (m, 3H)13C NMR(75MHz,CDCl3)δ196.18,172.89,
171.54,167.36,160.19,155.39,153.53,150.94,148.94,145.77,145.14,141.44,141.34,
139.87,137.77,136.46,135.68,132.54,132.44,130.73,130.61,130.46,130.40,129.43,
129.36,128.32,128.08,127.96,127.83,127.75,127.67,120.16,120.07,105.98,104.19,
95.76,92.70,78.21,77.36,72.98,69.86,67.14,66.64,62.38,51.59,50.06,48.12,
37.29,36.68,31.96,31.87,31.02,30.36,26.22,24.89,23.31,22.72,19.67,17.63,
14.12,12.22,11.50,11.16,7.82.
The ESI-MS that Fig. 1 is CPT-ss-HPPH is composed, and Fig. 2 is CPT-ss-HPPH's1H NMR spectras demonstrate preparation
Compound success.The uv-vis spectra of CPT-ss-HPPH also shows the characteristic spectrum (Fig. 5) of CPT and HPPH.
Embodiment 2：The preparation of CPT-cc-HPPH
DMAP (0.58mg, 4.75 μm of ol), HPPH (15mg, 23.5) and CPT-cc-OH are mixed in 20mL dichloromethane
(23.8mgmg, 47.1 μm of ol).Under stiring with the dichloromethane solution that 5mL EDC HCl (9.1mg, 47.1 μm of ol) are added dropwise.
After being stirred at room temperature 24 hours, confirm that HPPH reacts completely by thin-layer chromatography (TLC).It is concentrated and is mixed using rotary evaporator
Object solution then uses gradient ethyl acetate and hexane to pass through flash chromatography (Teledyne ISCO as eluent
CombiFlash it) purifies.Yield:22.5mg (86%) .ESI-MS m/z (M+)calcd 1110.55,found 1111.32(M
+H+).1H NMR(300MHz,CDCl3) δ 9.78 (d, J=4.1Hz, 1H), 9.52 (s, 2H), 8.50 (s, 1H), 8.19-7.95
(m, 4H), 7.79-7.59 (m, 2H), 7.57-7.45 (m, 2H), 7.27 (s, 2H), 5.98-5.83 (m, 2H), 5.68 (d, J=
17.3Hz, 2H), 5.37 (dd, J=17.1,1.1Hz, 2H), 5.28 (d, J=8.9Hz, 1H), 5.20 (s, 2H), 5.17-5.02
(m, 3H), 4.46 (q, J=7.4Hz, 1H), 4.28 (d, J=8.3Hz, 1H), 4.11-3.98 (m, 2H), 3.99-3.81 (m,
2H),3.79–3.53(m,7H),3.37(s,3H),3.27(s,3H),2.73–2.57(m,1H),2.56–2.39(m,1H),
2.38-2.15 (m, 2H), 2.11 (d, J=6.7,1.4Hz, 5H), 1.80 (d, J=7.3,1.3Hz, 4H), 1.71 (t, J=
7.6Hz, 5H), 1.57 (s, 6H), 1.42 (t, J=6.8Hz, 4H), 1.36-1.14 (m, 11H), 0.98 (t, J=7.5Hz,
3H),0.80–0.73(m,3H).13C NMR(75MHz,CDCl3)δ196.29,173.20,171.59,167.56,160.34,
157.38,155.39,153.91,152.35,150.96,149.10,146.47,145.92,145.13,141.50,139.86,
137.85,136.41,135.73,135.68,132.39,131.10,131.07,130.71,130.55,129.64,128.44
128.36,128.13,128.09,128.03,127.99,120.38,106.09,104.22,96.00,92.71,77.78,
77.36,72.94,69.84,69.06,67.19,64.55,51.75,50.12,49.96,48.18,32.06,31.86,
31.09,30.35,28.44,26.21,25.54,25.32,24.86,23.30,22.71,19.66,17.61,14.11,
12.23,11.49,11.19,7.78。
The ESI-MS that Fig. 3 is CPT-cc-HPPH is composed, and Fig. 4 is the 1H NMR spectras of CPT-cc-HPPH, demonstrates preparation
Compound success.
Embodiment 3：The active oxygen (ROS) of HPPH and CPT-ss-HPPH generates
It is prepared as ROS quenchers to measure HPPH and embodiment 1 by using anthracene -9,10- dipropionic acids (ADPA)
The ROS of CPT-ss-HPPH is generated.In brief, ADPA is dissolved in DMSO, is diluted at 410nm with 1.1 UV-
Vis absorbances.Then the HPPH or CPT-ss-HPPH that are added in 1mM DMSO simultaneously make final concentration of 10 μM.The solution obtained
It is irradiated 7 minutes with 671 nanometer lasers that intensity is 100mW.Per minute, by UV3100PC spectrophotometers (VWR,
Radnor, PA) recording solution UV-Vis absorbances.Acquired results are analyzed by Origin 8.
As shown in Figure 6, it has been found that with the increase of irradiation time, the absorbance of HPPH and CPT-ss-HPPH constantly subtract
It is small.It is worth noting that, compared with HPPH, the reduction of the absorbance of CPT-ss-HPPH faster, shows that CPT-ss-HPPH can
Generate more ROS (Fig. 6 a-c).In addition, the absorbance loss of HPPH or CPT-ss-HPPH shows that they are swashing at 662nm
Unstable (Fig. 6 a-b) during light irradiation.The quantitative analysis that loss by measuring absorbance at 662nm carries out is shown, is approached
90% HPPH degradations, in contrast, CPT-ss-HPPH is less than 20% (Fig. 6 d-e).These results indicate that by by CPT with
HPPH links together to form prodrug, and the photostability and ROS generative capacities of HPPH are improved.
Embodiment 4：The preparation of medicament nano particle
Nano particle is obtained by nanoprecipitation method.In brief, by CPT-ss-HPPH, CPT-cc-HPPH and HPPH with
The concentration of 100 μ g/mL is dissolved in tetrahydrofuran (THF), and adds different amounts of polymer P EG45-b-PLA28Respectively with it is upper
Several drugs are stated to be sufficiently mixed.Mixture solution is precipitated to from THF in water under stiring, and is steamed overnight in draught cupboard
Hair removes THF.Later, mixture is centrifuged 3 minutes to remove any possible sediment with 3000RPM.Existed by UV-Vis
660nm measures medicament contg.It is encapsulated about CPT, dissolves CPT and PEG using dimethyl sulfoxide (DMSO) (DMSO)45-b-PLA28, then
It is deposited in water dropwise under stiring.Due to the high aggregation in centrifugal process, cannot be centrifuged.Pass through high performance liquid chromatography
Method (HPLC) measures the amount of CPT.Drugloading rate (DL) and encapsulation rate (EE) are calculated according to following equation.
DL (%)=quality/carrier of entrapped drug and quality × 100% of entrapped drug
Weight × 100% of the weight of EE (%)=entrapped drug/initial intake drug
Result of study is as shown in table 1.Due to the intrinsic aroma properties of CPT, in aqueous solution and most of organic solvents all
With limited solubility.Dimethyl sulfoxide (DMSO) (DMSO) is the CPT uniquely usual vehicles with good solubility.In 4% medicine
Under object/polymer feed ratio, number of the nano particle with 430 ± 86nm obtained by the nanoprecipitation from DMSO to water flows
Body dynamics diameter (table 1, serial number 1).However, the nano particle obtained is unstable, and precipitated within several hours.As right
Than when the feed rate ratio of HPPH is 10%, load efficiency (LE) is up to 98% and drug loading (DL) is up to 8.9%
(table 1, serial number 2).When CPT is connect with HPPH, the hydrophilic hydroxy group of CPT and hydrophilic carboxylic acid's base of HPPH are separately converted to dredge
Aqueous carbonic ester and ester group cause the whole hydrophobicity of CPT-ss-HPPH to enhance.The hydrophobicity increase of CPT-ss-HPPH can
More effectively it is encapsulated as that there is than corresponding monomer higher LE and DL nano-carrier.The result shows that when drug/polymer ratio
Example from 0.2 increase to 1.5 when, the DL of CPT-ss-HPPH increases to from 16% close to 60%, and quantitative LE is 96% (table 1, serial number
3-6).Under 0.2 charge ratio, the hydrodynamic diameter of the nano particle obtained is 34 ± 9nm (Figure 10), has neutrality
Zeta potential (Figure 11).Transmission electron microscope (TEM) shows that these nano particles are spherical, and average-size is 35 ± 5nm (figures
7).It is found that similar Size Distribution when drug input ratio is 0.4.Under higher charge ratio, their diameter increases to
About 50nm (table 1, serial number 5-6).As expected, control CPT-cc-HPPH is under 0.2 drug/polymer feed rate ratio
Also there is high LE (table 1, serial number 7).Due to the simplicity and validity of CPT-ss-HPPH, excellent LE and DL grind conversion
Studying carefully has very high attraction.
Table 1
| Serial number | Drug | Ratioa | Diameter (nm)b | LE (%) | DL (%) |
| 1 | CPT | 0.04 | 430±86 | NAc | NAc |
| 2 | HPPH | 0.1 | 35±9 | 98 | 8.9 |
| 3 | CPT-ss-HPPH | 0.2 | 34±9 | 97 | 16 |
| 4 | CPT-ss-HPPH | 0.4 | 33±8 | 97 | 28 |
| 5 | CPT-ss-HPPH | 1.0 | 46±11 | 98 | 49 |
| 6d | CPT-ss-HPPH | 1.5 | 52±13 | 97 | 59 |
| 7 | CPT-cc-HPPH | 0.2 | 35±9 | 97 | 16 |
aThe charge proportion of drug/PEG-b-PLA；bThe digital averaging hydrodynamic diameter measured by DLS；cIt is unstable,
It is not purified；dVisible sediment is observed after 3 days.
Embodiment 5：Drug release
The drug release for being released through dialysis and measuring nano-particle of drug.In brief, phosphate buffered saline (PBS) is used
(PBS) drug concentration of nano-particle to 3.5 μM of the dilution comprising drug.1.0mL is transferred to semi-transparent membrane tube (MWCO：
In 50kDa), and dialyse 4 days in the 25mLPBS for containing or not contain 10mM glutathione (GSH) at 37 DEG C respectively.In difference
Time point, measure dialyzate in remaining drug.CPT-ss-HPPH, CPT-cc-HPPH and HPPH are measured by UV-Vis,
CPT is measured by HPLC.The sample solution of measurement is added back in dialyzate after measurement.Totally three groups of parallel tests.
Micellar drug is mainly diffusion controlled process from the release in macromolecule micelle preparation, this depends on several factors,
Intrinsic property including carrier and drug, distribution and release environment of the drug on carrier.In fact, one of micellar preparation
Common disadvantage is drug initial burst, this is typically drug-induced by the weak binding on nano grain surface.Ours
In CPT-ss-HPPH, lower water-soluble and increased molecular weight may cause drug releasing rate to be less than any list
Body drug.In addition, indeed it is contemplated that GSH can promote the drug release of CPT-ss-HPPH by cutting disulfide bond.Therefore, Wo Men
Had studied at 37 DEG C CPT NPs, HPPH NPs, CPT-cc-HPPH NPs and CPT-ss-HPPH in PBS drug release (
In the case of with and without 10mM GSH) (Fig. 8).As expected, CPT and HPPH from the release in nano particle very
Soon, most of micellar preparations are similar to, release half-life period is only 4.6 hours and 5.6 hours respectively.In contrast, CPT-ss-
HPPH and CPT-cc-HPPH shows that almost Zero order release, wherein drug are almost discharged with constant rate of speed.Their elimination half life values quilt
It is determined as 51 hours (CPT-ss-HPPH) and 46 hours (CPT-cc-HPPH), this is about 10 times than any monomer.For drug
For delivering, slow Zero order release is in demand, because it can reduce the release of the premature drug during blood circulation,
And therefore improve side effect.As described above, another important feature of CPT-ss-HPPH is the disulfide bond of its GSH sensitivities, permit
Perhaps it HPPH and is discharged by two step cascade reactions without the CPT of modification.Our observation confirms this feature, i.e. 10mM
GSH causes drug release faster really, release half-life period only 24 hours (Fig. 8).There are GSH, release partly declines
Significant reduction (24 hours to 51 hours) will allow effective pharmacological activation and release after changing in the cell phase, thus allow CPT-
Ss-HPPH effectively kills cell.These drug release studies show that the CPT-ss-HPPH nano particles of our GSH sensitivities are
One response medicine release system well controlled.
Embodiment 6：Vitro cytotoxicity measures
Human colon cancer cell HCT116 is layered on the 96 of McCoy's 5a culture mediums (10% fetal calf serum and 1% penicillin)
In orifice plate.Cell is contained into 5%CO at 37 DEG C2Humid atmosphere in incubation.By drug or it is loaded with medicine using cell culture medium
The nano particle of object is diluted to predetermined concentration.100 μ L are added per hole has the cell culture medium of different specific drug concentration.Pass through
100 μ L culture mediums are added and generate negative control.After 24 hours, HPPH, CPT-ss- are contained with 10mW irradiations with 671nm laser
The sample well of HPPH or CPT-cc-HPPH 1 minute.After irradiation, these cells are incubated 24 hours again.For not laser treatment
Cell is directly incubated 48 hours by sample.After incubation, with the 3- (4,5- dimethylthiazole -2- bases) -2,5- containing 0.5mg/mL
100 μ L culture mediums of diphenyltetrazoliumbromide (MTT) are replaced culture medium and are therewith incubated 2 hours.It removes containing unreacted
After the culture medium of MTT, the blue first crystals crystallizations on the left side are dissolved in 100 μ LDMSO, in BioTek Synergy H4
Absorbance is measured with wavelength 570nm in hybrid readings instrument.The absorbance measured subtracts blank control, and is based on not locating control
The Relative Absorbance of the cell of reason calculates cell viability.Calculating and the statistics of IC50 values are carried out using GraphPad Prism 5
Analysis.
We measure free CPT, HPPH and their 1 first:The cytotoxicity of 1 mixture.The IC of HPPH and CPT50
Respectively 92nM and 100nM, and the IC combined50Drop to 23nM (Fig. 9).Medication combined index (CI) is calculated as 0.49, this table
There is high concertedness between bright CPT and HPPH.Powerful synergistic effect, which may be their different cell killing mechanism and CPT, to be made
Cell is obtained easily by the synthesis result of the low oxygen level caused by laser irradiation HPPH.Due to relatively slow release, CPT-ss-
Vitro cytotoxicity (ICs 50 of the HPPH NPs in no laser irradiation：4.9 μM) show obtain lower than any monomer medicine
It is more, and non-response property CPT-cc-HPPH NPs show low-down vitro cytotoxicity (IC50>100 μM), show disulfide bond
Key effect in activating CPT-ss-HPPH therapeutic efficiencies.Laser irradiation with the cell of CPT-ss-HPPH NPs processing is aobvious
Write enhancing cytotoxicity, IC50It is reduced to 1.4 μM from 4.9 μM.If we assume that the cell of the induced with laser of CPT-ss-HPPH
Toxicity is close to CPT-cc-HPPH (IC50：34 μM), then the CI of CPT and HPPH will be 0.33 in CPT-ss-HPPH.
Embodiment 7：Cell in vitro absorbs
We are external using fluorescence microscope research HPPH NPs, CPT-ss-HPPH NPs and CPT-cc-HPPH NPs'
Cellular uptake.5 μM each preparation and HCT116 cells are incubated with 24 hours, and used at 37 DEG C before fluorescence imaging
Hoechst33342 (Life Technologies, Carlsbad, CA) is dyed 0.5 hour in cell culture couveuse.Then
Before imaging cell is washed with Dulbecco's PBS three times.The channels selection DAPI are used to detect Hoesse33342 dyeing
Nucleus selects the channels CY7.5 to collect the fluorescence from HPPH, CPT-ss-HPPH and CPT-cc-HPPH.As a result such as Figure 12 institutes
Show, CPT-ss-HPPH NPs and CPT-cc-HPPH NPs are effectively internalized by cell as HPPH NP.
Embodiment 8：Radioactivity [64Cu] label HPPH NPs and CPT-ss-HPPH NPs and they in HCT116 tumours
Internal PET imaging
CPT-ss-HPPH and HPPH are dissolved in 1.0mg/mL in DMSO respectively.Then, 0.1mL is incubated at 50 DEG C, and
With 0.1mL in 0.4M sodium acetates (pH=5.6) buffer solution [64CuOAc] mixing.Mixture solution is further incubated for 5-7
Minute, 0.2mL PEG are then added45-b-PLA28(10mg/mL in DMSO).After futher stirring 30 minutes, 0.2mL water is added
Enter into mixture, is then purified using PBS as eluent by PD-10 columns.Total label program flower including purifying
About 1 hour is taken.Before intravenous injection, radiolabeled HPPH NPs and CPT- β-HPPH NPs and PEG45-b-PLA28
Stock solution mixing, it is about 10mg/mL to make the ultimate density that polymer obtains, and radioactivity is 120-150 μ Ci (4.44-
5.55MBq)/100μL。
By being subcutaneously injected 5 × 106Suspension of the HCT116 cells in PBS (100 μ L) prepares HCT116 tumor bearing nude mices
(7 week old, female).When tumor size reaches 500mm3When, mouse is imaged for PET.Before tracer injection, different fluorine is used
Alkane/O 2 (2%v/v) anesthetized mice.Above-mentioned PEG45-b-PLA28With64The drug of Cu labels is quiet after mixing in PBS (100 μ L)
The mouse (every mouse 4.44-5.55MBq/120-150 μ Ci) of arteries and veins injecting anesthetic.Specified time point after injection,
Mouse is scanned on Inveon DPET scanners.PET image is rebuild using 3D order subset expectation-maximization algorithms to decline without correcting
Subtract or scatters.ASI Pro VMTM softwares are used for image analysis.Area-of-interest (ROI) is drawn using IRW (Siemens).Injection
Put to death above-mentioned mouse within 48 hours afterwards.It collects organ and weighs weight in wet base.γ-counter (Wallac Wizard 1480,
PerkinElmer the organ and series of standards solution of collection are measured on)64Cu radioactivity.The radioactivity of organ is converted in terms of
Calculate the percentage (%ID/g) of per gram of tissue injection dosage.
As shown in figure 13, it compared with HPPH, is dramatically increased in the CPT-ss-HPPH that corresponding time point is accumulated in tumour.
Quantization by the PET image of decay correction show the tumor accumulation of CPT-ss-HPPH reach within 24 hours 6.0 after injection ±
0.6%ID/g and it was maintained for up to 6.1 ± 0.8%ID/g at 48 hours, and HPPH is at 48 hours only 2.8 ± 0.8%
ID/g (Figure 14).In vitro bio distribution based on the organ progress γ countings to excision confirms that CPT-ss-HPPH is significant and is higher than
The tumor accumulation (Figure 15) of HPPH.Since the nano-carrier size of HPPH and CPT-ss-HPPH is identical, CPT-ss-HPPH is opposite
The too early of CPT-ss-HPPH during blood circulation is attributed in the significant higher accumulation of HPPH to discharge less, thus in tumour
Middle reservation is higher.It should be noted that the slow too early release of CPT-ss-HPPH is it is also contemplated that complete after reducing intravenous administration
Body toxicity.
Embodiment 9：Interior therapeutic HCT116 tumor-bearing mices
By being subcutaneously injected 5 × 106Suspension of the HCT116 cells in PBS (100 μ L) prepares HCT116 tumor bearing nude mices
(7 week old, female).The 10th day (about 60mm after tumour is in tumor inoculation3) when, mouse starts with by being injected intravenously 100 μ
L drugs or PBS, every 4 days primary 2 (CPT dosage in total：3.0mg/kg, HPPH：5.5mg/kg, CPT-ss-HPPH：9.9mg/
Kg, CPT-cc-HPPH：9.6mg/kg).2 days after per injection, with the 671nm laser irradiations tumour 10 minutes of 200mW.Every 2
Its monitoring gross tumor volume and mouse weight.When the tumour of any size close to 2 centimetres or when mouse weight lose more than 20% when,
Mouse is set to be euthanized.The volume of each tumour is calculated separately using following formula：
Volume=(length * width 2)/2
Use GraphPad Prism 5 (La Jolla, CA) analysis result.
Since PET is imaged the tumor accumulation highest of CPT-ss-HPPH after display is injected 48 hours, we are every 4 days to small
Two doses of mouse intravenous administration, and after per injection 2 days using 671nm laser irradiations tumour 10 minutes, in 200 milliwatts/flat
Square centimetre.As shown in figure 16, it with the tumour fast-growth of the mouse of PBS or PBS+ laser treatments, but is handled with laser irradiation
CPT-ss-HPPH NPs have been reduced significantly gross tumor volume.On the contrary, CPT NPs and HPPH NPs show limited tumour life
It is long to inhibit.In addition, CPT-cc-HPPH clearly demonstrates that GSH can relative to the apparent poor tumor suppression of CPT-ss-HPPH
The necessity of the disulfide bond of cutting.The above results consistently show that the mouse with CPT-ss-HPPH NPs processing shows than it
His best survival rate of processing group (Figure 17).
In short, we devise a kind of completely new GSH response multi-functional prodrugs of heterodimer, it can effectively be loaded into polymerization
In object nano-carrier, and two kinds of cooperative drugs are delivered to tumour jointly, so as to cause advantageous oncotherapy.It is of the present invention
The multi-functional prodrug of heterodimer can be prepared by simply reacting, and with high load capability and quantitative load efficiency packet
In enclosed biocompatibility PEG-b-PLA.It is worth noting that, receiving of being formed of preferred prodrug CPT-ss-HPPH in the present invention
Rice grain shows to discharge slow 10 times than the nanoparticulate drug for loading monomer medicine accordingly；Disulfide bond in its structure for
On-demand drug release and the effective cytotoxicity of holding are essential.In addition, photoluminescent property and HPPH can directly be put
The ability of penetrating property label allows us, and by fluorescence microscope, its cell in vitro absorbs drug, and passes through PET imaging research
Its internal drug imaging, it was demonstrated that the nano particle of load C PT-ss-HPPH significantly improves the drug delivery to tumour.Most
Afterwards, CPT-ss-HPPH nano particles of the invention show that the tumour of collaboration is controlled by combination chemotherapy and photodynamic therapy
Treat effect.In general, our multi-functional pro-drugs are that drug delivery opens new approach.
Claims (10)
1. a kind of multi-functional prodrug of heterodimer based on camptothecin, it is loaded with camptothecin and 2- (1- hexyloxy second simultaneously
Base) -2- goes the dilute Ji-Jiao Tuo of second to delay chlorophyllin-a (HPPH), and it is the camptothecine that camptothecin is formed with HPPH by linking group
Class-HPPH prodrugs, shown in structural formula such as formula (I)：
Wherein,
R is-CH2CH2,-SS- orIn one kind；R1 is camptothecin group.
2. multi-functional prodrug described in claim 1, shown in structure such as following formula (II)
3. the method for preparing multi-functional prodrug described in claim 1, specifically includes following steps：
1) in organic solvent, using on triphosgene activation camptothecine or derivatives thereof lactonic ring in the presence of acylation catalyst
Hydroxyl adds bis- thiodiethanols of excessive 2,2'- or 1, and 6- hexylene glycols reaction or 2, (propane -2,2- diyl is double by 2'-
(sulphur)) diethanol, obtain intermediate 1；
2) intermediate 1 and HPPH that step 1) obtains are mixed in organic solvent, the ester bond generated by coupling reagent reaction,
Obtain the part camptothecin-HPPH prodrugs.
4. the method described in claim 3, it is characterised in that：Organic solvent described in step 1) is selected from dichloromethane, chloroform, four
Any one in hydrogen furans, 1,4- dioxane or dimethylformamide；Acylation catalyst described in step 1) is selected from N- second
Base-N '-[(3- dimethylaminos) propyl] carbodiimide hydrochloride (EDC), N, N'- dicyclohexylcarbodiimides (DCC) it is arbitrary
It is a kind of；Organic solvent described in step 2) is selected from dichloromethane, chloroform, tetrahydrofuran, 1,4- dioxane or dimethyl formyl
Any one in amine；Coupling reagent described in step 2) is selected from 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne diimmonium salt
Hydrochlorate (EDC.HCL), N, N- dicyclohexyl carbonic acid diimines (DCC) or 1H- benzotriazole -1- base oxygen tripyrrole alkyl hexafluoro phosphorus
Any one in hydrochlorate (PyBOP).
5. multi-functional prodrug described in claim 1 is in pharmaceutically acceptable preparation.
6. the preparation described in claim 5, it is characterised in that：The preparation is nanoparticle formulations, the nano particle
Outside is excipients and/or pharmaceutical carrier, and happiness described in claim 1 is wrapped up inside the excipients and/or pharmaceutical carrier
Set bases-HPPH prodrugs.
7. the preparation described in claim 6, it is characterised in that：The excipients and/or pharmaceutical carrier are amphipathy macromolecule；
It is preferred that polyethylene-b-polylactic acid (polyethylene glycol-b-polylactide, PEG-b-PLA), polyethylene glycol-
B- polycaprolactones (polyethylene glycol-b-polycaprolactone, PEG-b-PCL) or the poly- phosphorus of polyethylene glycol-b-
Any one in ester (polyethylene glycol-b-polyphosphoester, PEG-b-PPE).
8. application of the multi-functional prodrug described in claim 1 in preparing cancer treatment drugs.
9. application according to any one of claims 8, it is characterised in that：Using amphipathy macromolecule to described in claim 1 multi-functional
Prodrug carries out physically encapsulation, and nano particle is prepared.
10. the application described in claim 9, it is characterised in that：By camptothecin-HPPH prodrugs described in claim 1 and two
Parent's property macromolecule is dissolved in organic solvent together, then instill stirring or ultrasound aqueous solution in, then vapor away organic
Nano particle is made in solvent；The amphipathy macromolecule appointing in PEG-b-PLA, PEG-b-PCL or PEG-b-PPE
Meaning is a kind of.
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| WO2021005583A1 (en) | 2019-07-11 | 2021-01-14 | Sun Pharma Advanced Research Company Ltd. | Camptothecin derivatives with a disulfide moiety and a piperazine moiety |
| CN111358948A (en) * | 2020-03-31 | 2020-07-03 | 东南大学 | Camptothecin-berberine/indocyanine green nano-drug, preparation method and application |
| CN112047952A (en) * | 2020-08-28 | 2020-12-08 | 四川大学 | Camptothecin-photosensitizer prodrug and preparation method and application thereof |
| CN112047952B (en) * | 2020-08-28 | 2022-04-29 | 四川大学 | Camptothecin-photosensitizer prodrug and preparation method and application thereof |
| CN112263566A (en) * | 2020-09-24 | 2021-01-26 | 中国药科大学 | Albumin-binding type anoxic-oxidation dual-responsiveness composite nanoparticle, preparation method and application |
| CN112263566B (en) * | 2020-09-24 | 2022-06-24 | 中国药科大学 | Albumin-binding type anoxic-oxidation dual-responsiveness composite nanoparticle, preparation method and application |
| CN112321615A (en) * | 2020-10-30 | 2021-02-05 | 华中科技大学 | Camptothecin-based dimer compound, and preparation and application thereof |
| CN112321615B (en) * | 2020-10-30 | 2021-11-09 | 华中科技大学 | Camptothecin-based dimer compound, and preparation and application thereof |
| WO2022088679A1 (en) * | 2020-10-30 | 2022-05-05 | 华中科技大学 | Method for removing tumor stem cells, anti-cancer drug, drug delivery system, and use thereof |
| CN112472683A (en) * | 2020-11-11 | 2021-03-12 | 深圳大学 | Nano diagnosis and treatment agent and preparation method and application thereof |
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