CN114306620A - Human serum albumin nano-drug based on metabolism check point and preparation method and application thereof - Google Patents
Human serum albumin nano-drug based on metabolism check point and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a human serum albumin nano-drugs (IDNPs) based on metabolic check points and a preparation method and application thereof. The prodrug molecule IDOi-DON obtained by coupling indoleamine-2,3-dioxygenase inhibitor (IDOi) and metabolic reprogramming agent DON through chemical bond amidation reaction can be subjected to enzymolysis and fracture in a tumor microenvironment, and is coated with natural biocompatible serum albumin, so that the stability of the drug can be improved, and the toxic and side effects of the drug are reducedThe enrichment of tumor part can inhibit IDO activity of tumor cells, inhibit Treg cell proliferation and activate CD8 through the synergistic effect of the two medicines+The T cell function synergistically enhances the anti-tumor immunotherapy effect.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to a human serum albumin nano-drug based on a metabolism check point and a preparation method and application thereof.
Background
Tumor immunotherapy kills or inhibits tumor growth by stimulating the body's anti-tumor immune response, a revolutionary breakthrough in the tumor field in recent years. Still, an important problem is that immune tolerance formed by a tumor microenvironment cannot be overcome, and the immune microenvironment formed by vigorous tumor metabolism inhibits the metabolism of immune cells, so that the tumor cells cannot be effectively identified and killed. Therefore, aiming at the metabolic target of tumor immunosuppression, the functional metabolic examination regulator for inducing controllable immune response is developed, and the preparation method has important significance for cancer immunotherapy.
Tumor immunotherapy achieves inhibition of tumor progression by activating immune cells to recognize and eliminate tumor cells. The tumor immunosuppressive microenvironment is a key factor causing tumor growth and metastasis, and various immunosuppressive cells in the tumor microenvironment can inhibit antitumor immune responses, such as interleukin-10 (IL-10), indoleamine 2,3-dioxygenase (IDO), and the like. Immunosuppressive factors in the tumor microenvironment severely inhibit activation, migration and differentiation of immune cells, limiting the responsiveness of anti-tumor immunotherapy. The IDO inhibitors such as 1-MT, NLG919 and the like can inhibit the degradation of tryptophan, twist dendritic cells to inhibit T cells, activate the T cells to generate immune response and finally inhibit or eliminate tumors. However, due to the complexity of the immunosuppressive microenvironment, it is difficult to reactivate T cells by inhibiting only one immunosuppressive pathway, and IDOi shows weak antitumor effect and significant drug side effects in clinical studies. Therefore, there is a need to design agents or molecules that target multiple targets simultaneously to improve the immunotherapeutic effect.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a human serum albumin nano-drug based on a metabolic checkpoint and a preparation method and application thereof. Coupling indoleamine-2,3-dioxygenase inhibitor (IDoi) and metabolic reprogramming agent DON through chemical bond amidation reaction to obtain prodrug molecule IDoi-DON, and coating natural biocompatible human serum albumin outside to obtain the human serum albumin nano-drug based on the metabolic checkpoint.
The invention provides a human serum albumin nano-drug based on a metabolism check point, which comprises a prodrug molecule inner core and human serum albumin coated outside, wherein the prodrug molecule is a coupling molecule of indoleamine-2,3-dioxygenase inhibitor and a metabolism reprogramming agent, and the coupling mode is amido bond coupling.
Further, the metabolic reprogramming agent is a glutamine antagonist.
Further, the metabolic reprogramming agent is any one selected from 6-diazo-5-oxo-norleucine, 5-diazo-4-oxo-L-norvaline, L- (alpha-S, 5S) -alpha-amino-3-chloro-4, 5-dihydro-5-isoxazoleacetic acid and azaserine, and the connection principle is the same.
Further, the indoleamine-2,3-dioxygenase inhibitor is an immune activator.
Further, the immune activator is 1-methyltryptophan.
Further, the human serum albumin coats the inner core of the prodrug molecule through hydrophobic interaction.
Further, the structure of the prodrug molecule inner core is shown as the formula (I):
wherein n is any one of 0, 1, 2,3, 4,5, 6 and 7; r1And R2Selected from H or alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl; r3Is H or can be adjacent to R3The nitrogen forms an amide bond, a carbamate bond, a phosphoramidate bond, a phosphorodiamidite bond; r3' is selected from H or C1-C8 alkyl, substituted C1-C8 alkyl, or R3And R3Together,' form a ring structure comprising-C (═ O) -G-C (═ O) -, where G is selected from the group consisting of: C1-C8 alkylene, C1-C8 heteroalkylene, C5-C8 cycloalkylene, C6-C12 arylene, C5-C12 heteroarylene, divalent C4-C10 heterocycle, each of which may be optionally substituted.
The invention also provides a preparation method of the human serum albumin nano-drug based on the metabolic checkpoint, which comprises the following steps:
(1) synthesis of 9-fluorenylmethoxycarbonyl protected (indoleamine-2,3-dioxygenase inhibitor) -metabolic reprogramming agents;
(2) removal of the 9-fluorenylmethoxy carbonyl group to produce a (indoleamine-2,3-dioxygenase inhibitor) -metabolic reprogramming agent prodrug molecule;
(3) preparation of (indoleamine-2,3-dioxygenase inhibitor) -metabolism reprogramming agent human serum albumin nano-drug.
Further, in the step (1), the isopropyl-metabolic reprogramming agent and the 9-fluorenylmethoxycarbonyl-indoleamine-2, 3-dioxygenase inhibitor are coupled to generate the 9-fluorenylmethoxycarbonyl-protected indoleamine-2,3-dioxygenase inhibitor-metabolic reprogramming agent in the presence of benzotriazole-N, N, N ', N' -tetramethyluronium hexafluorophosphate and diisopropylethylamine.
Further, in the step (2), the product obtained in the step (1) reacts overnight under the action of piperidine to remove the carbonyl group of 9-fluorenylmethoxy group, so as to obtain the prodrug molecule.
Further, in the step (3), (1) the prodrug molecule obtained in the step (2) is dissolved in an organic solvent, and under the continuous action of external force, the prodrug molecule solution is slowly dripped into the aqueous solution of the human serum albumin to induce the self-assembly of the human serum albumin molecules, and the human serum albumin nano-drug based on the metabolic checkpoint is obtained through purification and filtration.
The invention also provides application of the human serum albumin nano-drug based on the metabolic checkpoint in tumor immunotherapy.
In summary, compared with the prior art, the invention achieves the following technical effects:
(1) the prodrug molecule is prepared by chemically coupling the IDOi and the metabolic reprogramming agent, and can be broken into two acting drug molecules by enzymolysis in a tumor microenvironment, so that tumor cells are subjected to space-time synergistic action, and the tumor growth is inhibited.
(2) The invention utilizes the hydrophobic drug-induced human serum albumin molecule self-assembly technology to prepare the IDoi-DON albumin nano-drugs (IDNTPs), can improve the stability of the drugs, reduce the toxic and side effects of the drugs, improve the enrichment of tumor parts, improve the tumor treatment efficiency, have good biological safety, and overcome the toxicity challenge of IDoi and DON in clinical application.
(3) The IDOi inhibits Treg cell proliferation and DON activates CD8+The T cells synergistically enhance the anti-tumor immune response and effectively improve the immunotherapy effect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 shows the hydrogen nuclear magnetic resonance spectrum of IDOi-DON.
FIG. 2 is a mass spectrum of IDoi-DON.
FIG. 3 is a particle size distribution and transmission electron microscope image of IDNPs.
FIG. 4 is a growth inhibition evaluation of SPCA1 lung cancer cells by IDNPs.
Detailed Description
In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, shall fall within the scope of protection of the present invention.
Tryptophane metabolites mediated by Indoleamine-2,3-dioxygenase (IDO) are important physiological substances for anti-tumor immunosuppression, and therefore IDO is an important metabolic target (metabolic checkpoint) for tumor immunotherapy. IDO has the capability of catalyzing the degradation of tryptophan, and kynurenine which is a metabolite mediated by IDO can promote the generation of new blood vessels and inhibit the function of immune cells, thereby escaping the monitoring and removing capability of an organism on tumor cells. IDO is highly expressed in tumors and tumor metastatic lymph nodes and is involved in immune tolerance mechanisms of tumors. IDO inhibitors (IDO inhibitors, IDO) represented by 1-methyltryptophan (1-MT) can inhibit the growth of tumor cells by enhancing the anti-tumor immune response. However, in preclinical studies, IDOi only exhibits moderate in vivo anti-tumor activity, and co-administration is limited by complex pharmacokinetics and drug-drug interactions, and therefore there is a need to design one molecule that can target multiple metabolic checkpoints, overcoming these inherent challenges.
Metabolic reprogramming is also a hallmark of malignancy, cancer progression is dependent on metabolic reprogramming, and inhibition of certain metabolic pathways in tumors is also an important approach to cancer treatment. Glutamine metabolism is also closely related to tumor growth and survival, glutamine generates glutamic acid under the action of glutaminase, then the glutamic acid is converted into alpha-ketoglutaric acid, enters tricarboxylic acid (TCA) cycle, and is used for synthetic generationThank you play an important role. A structural analog of glutamine, 6-diazo-5-oxo-L-norleucine (DON), can inhibit the activity of enzyme with glutamine as substrate very broadly and effectively, and DON blocks glutamine metabolism, can inhibit the oxidative phosphorylation of tumor cells, and can promote CD8+Proliferation and activation of T cells, in turn, act synergistically with IDOi to ensure the development of a strong, sustained immune response. However, oxidative phosphorylation metabolic pathway is elevated in T cells and can activate CD8+T cells, enhancing anti-tumor immunotherapy. However, DON is limited in its use by terminating in phase II clinical trials due to its toxicity. The simple combined drug can face the complex pharmacokinetics problem and can not avoid the side effect of the drug, so the chemically coupled IDOi and DON is a feasible method which can target two immune metabolic pathways, inhibit the metabolism of tumor cells, enhance the activity of T cells and weaken the side effect of the drug, and is expected to comprehensively improve the inhibition of the tumor growth.
The preparation of nanoparticles by coating Human Serum Albumin (HSA) can increase the bioavailability of the drug, and increase the enrichment of the nano-drug in tumor sites by utilizing the biocompatibility of HSA and the passive targeting of the Enhanced Permeability and Retention (EPR) effect of solid tumors, and can overcome the challenge of clinical toxicity.
The rapid development of nanotechnology has led to a dramatic development in the targeted therapy and in vivo diagnosis of tumors. The nanometer material is used as a medicine carrier, can improve the stability of the medicine, reduce the toxic and side effect of the medicine, ensure that the medicine is accurately released at a tumor part or achieves the effects of slow release and controlled release, and overcome the defects of low bioavailability, serious adverse reaction and the like of the traditional medicine.
Based on the reasons, the invention designs a protein nano-drug encapsulating metabolic reprogramming agent-immune activator prodrug, and the mechanism of the anti-tumor effect of the protein nano-drug is as follows: the inner core of the nano-drug with the anti-tumor effect is an IDOi-DON prodrug molecule with double metabolic targets. The prodrug molecule IDOi-DON is obtained by amidation coupling and deprotection of IDOi and DON. By chemical coupling of IDoi and metabolic reprogrammingThe prodrug molecule prepared by DON can be broken in the tumor microenvironment in response to enzyme, natural biocompatible serum albumin is coated outside the prodrug molecule, and the functional protein nano-drug can be enriched at the tumor part, inhibit the oxidative phosphorylation of tumor cells, inhibit the proliferation of Treg cells and activate CD8 at the same time+T cells, enhancing the anti-tumor immunotherapy effect.
The nano prodrug based on the metabolism check point is a double-drug-coupled prodrug molecule and is a prodrug molecule (IDOi-DON) coupled by a metabolism reprogramming agent-immune activating agent.
The metabolic reprogramming agent-immune activator coupled prodrug molecule (IDOi-DON) has the structure of formula (I).
The metabolic reprogramming agent is a glutamine antagonist.
The metabolic reprogramming agent is any one selected from 6-diazo-5-oxo-norleucine (DON), 5-diazo-4-oxo-L-norvaline (L-DONV), acivicin (acivicin) (L- (alpha-S, 5S) -alpha-amino-3-chloro-4, 5-dihydro-5-isoxazoleacetic acid) and azaserine.
The immune activator is indoleamine-2,3-dioxygenase inhibitor (IDOi) 1-methyltryptophan.
The preparation method of the double-drug conjugate prodrug molecule comprises the following steps: coupling 1-methyltryptophan (1-MT) protected by Fmoc group and isopropyl-DON through amido bond, and separating the obtained coupled molecule from the Fmoc group through hydrolysis reaction to obtain the double-drug coupled prodrug molecule, which specifically comprises the following steps:
(1) isopropyl-DON and 9-fluorenylmethoxycarbonyl-1-methyltryptophan are coupled in the presence of benzotriazole-N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HBTU) and diisopropylethylamine to generate Fmoc-protected IDOi-DON;
(2) carrying out overnight reaction on the Fmoc-protected IDOi-DON obtained in the step (1) under the action of piperidine to remove an Fmoc protecting group to obtain a target product IDOi-DON;
the invention also provides a preparation method of human serum albumin nano-drugs (IDNPs) based on the thank eximer checkpoint-immunoactivator prodrug, and the IDOi-DON hydrophobicity is utilized to induce the self-assembly of Human Serum Albumin (HSA) molecules to prepare the functional protein nano-drugs.
According to the preparation method of the protein nano-drug, the obtained IDOi-DON molecules have hydrophobicity, and can induce albumin molecules to self-assemble under the action of external force, so that the IDNTPs functional protein nano-drug is prepared. The method comprises the following steps:
(1) dissolving IDOi-DON molecules obtained by chemical synthesis in a small amount of organic solvent, slowly dropping IDOi-DON solution into HSA aqueous solution under the continuous action of external force such as ultrasound, stirring and vortex oscillation, and inducing albumin molecule self-assembly. The organic solvent includes a volatile organic solvent and a non-volatile organic solvent.
(2) And purifying the nanoparticle solution obtained in the first step, removing free drug molecules to obtain a functional protein nanoparticle solution, and filtering the functional protein nanoparticle solution through a filter membrane of 220nm to obtain the IDNPs functional protein nanoparticle solution. The purification means comprises dialysis, ultrafiltration and centrifugation.
The invention also provides a verification experiment of albumin nano-drugs (IDNPs) of IDoi-DON in vitro for anti-tumor treatment, namely CD19 in vitro+In the SPCA1 tumor model, tumor cells and IDNTPs nanoparticles are incubated for 72 hours, and under the action of glutathione in the tumor cells, the IDNTPs are decomposed into IDoi and DON to kill the tumor cells cooperatively.
The indoleamine-2,3-dioxygenase inhibitor in the following examples is 1-methyltryptophan (1-MT) as an example, and the metabolic reprogramming agent is 6-diazo-5-oxo-L-norleucine (DON) as an example, to show the preparation method of the human serum albumin nano-drugs (IDNPs) based on the metabolic checkpoint, and since the indoleamine-2,3-dioxygenase inhibitors have similar structures and the metabolic reprogramming agents also have similar structures, those skilled in the art can deduce that other indoleamine-2,3-dioxygenase inhibitors and metabolic reprogramming agents can be used to prepare the nano-drugs according to the method in the following examples.
EXAMPLE 19 Synthesis of fluorenylmethoxycarbonyl protected IDOi-DON
The preparation method comprises the following steps:
880mg of 9-fluorenylmethoxycarbonyl-1-methyltryptophan (2.06mmol, 1.1 equiv.) and 854mg of HBTU (2.25mmol, 1.2 equiv.) were weighed out and dissolved in 14mL of dry N, N-Dimethylformamide (DMF), and 980. mu.L of a solution of diisopropylethylamine (727mg, 5.63mmol, 3 equiv.) and isopropyl-DON (400mg, 1.88mmol, 1 equiv.) in dry DMF were added in this order and stirred under an inert gas atmosphere for 4 hours.
DMF was removed by rotary evaporation, 100mL of Dichloromethane (DCM) was added, and the resulting organic solutions were successively washed with 100mL of saturated sodium bicarbonate (NaHCO)3) The solution, water, 1mol/L hydrochloric acid (HCl), water and a saturated sodium chloride (NaCl) solution were washed, and dried over anhydrous magnesium sulfate. After DCM column chromatography (separation solvent: dichloromethane/ethyl acetate 5/1) was evaporated, the product was isolated and purified as a pale yellow solid, i.e. 9-fluorenylmethoxycarbonyl-IDOi-DON, the synthetic route is shown below.
EXAMPLE 2 preparation of IDOi-DON by removal of the 9-fluorenylmethoxy-carbonyl group
9-Fluorovinylmethoxycarbonyl-IDOi-DON (900mg, 2.07mmol, 1 equiv.) was weighed out and dissolved in 10mL of dry DCM, piperidine (514. mu.L, 5.17mmol, 2.5 equiv.) was slowly added dropwise, and stirring was continued at room temperature for 4 hours.
The organic solvent was removed by rotary evaporation and vacuum pump, and purified by column chromatography (separation solvent: chloroform/methanol: 30/1) to give IDOi-DON as a yellow product, as shown in the following scheme.
The obtained IDOi-DON was characterized, and its hydrogen nuclear magnetic resonance spectrum is shown in FIG. 1, in which the peak at-3.7 ppm belongs to-CH on the indole ring3The peak at-1.3 ppm is attributed to the two-CHs of the isopropyl group3. The mass spectrum of IDOi-DON is shown in FIG. 2, where 414m/z is assigned to the protonated peak of the produced IDOi-DON.
Example 3 preparation of human serum albumin nano-drugs (IDNPs) of IDOi-DON specifically includes the following steps:
(1) dissolving the chemically synthesized IDOi-DON molecules in a small amount of organic solvent, and subjecting an aqueous solution of Human Serum Albumin (HSA) to sonication with a VCX130 sonicator for 5min while dropwise adding the IDOi-DON solution to induce self-assembly of albumin molecules.
(2) Transferring the solution obtained in the step (1) into a 10kDa ultrafiltration tube, centrifuging, washing with deionized water for multiple times to remove free drug molecules, and filtering the functional protein nanoparticle solution with a filter membrane of 220nm to obtain the functional protein nano-drug solution of IDNPs.
In the embodiment, the nano particles are characterized and identified, as shown in fig. 3, the IDNPs transmission electron microscope shows a complete spherical structure, and the surface particle size of the dynamic light scattering detection result is about 100nm, which accords with the electron microscope result.
Example 4 killing of tumor cells by IDNPs
Will be 1 × 106Cultured SPCA1 cells were plated in 96-well plates at 37 ℃ in 5% CO2The cells were incubated for 24 hours under the conditions of (1) and (2) mM Glutathione (GSH) and different concentrations of IDNTPs were added in groups, and after further incubation for 72 hours, cell proliferation was detected using a CCK-8 kit. As shown in FIG. 4, under the action of GSH, the nano-drugs of IDNTPs show growth inhibition on tumor cells, and the inhibition efficiency is in direct proportion to the concentration of IDoi-DON in the IDNTPs.
The invention discloses a human serum albumin nano-drugs (IDNPs) based on metabolic check points and a preparation method and application thereof. The prodrug molecule IDOi-DON obtained by coupling indoleamine-2,3-dioxygenase inhibitor (IDOi) and metabolic reprogramming agent DON through chemical bond amidation reaction can be subjected to enzymolysis and breakage in a tumor microenvironment, natural biocompatible serum albumin is coated outside the prodrug molecule IDOi-DON, the stability of the drug can be improved, the toxic and side effects of the drug are reduced, the prodrug molecule IDOi-DON is enriched at a tumor part, and the effects of inhibiting IDO activity of tumor cells, inhibiting Treg cell proliferation and activating CD8 can be achieved through the synergistic effect of the two drugs+The T cell function synergistically enhances the anti-tumor immunotherapy effect.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (12)
1. The human serum albumin nano-drug based on the metabolic checkpoint is characterized by comprising a prodrug molecule inner core and human serum albumin coated outside, wherein the prodrug molecule is a coupling molecule of indoleamine-2,3-dioxygenase inhibitor and metabolic reprogramming agent, and the coupling mode is amido bond coupling.
2. The metabolic checkpoint-based human serum albumin nanopharmaceutical according to claim 1, wherein the metabolic reprogramming agent is a glutamine antagonist.
3. A human serum albumin nano-drug based on metabolic checkpoint as claimed in claim 2, wherein the metabolic reprogramming agent is selected from any one of 6-diazo-5-oxo-norleucine, 5-diazo-4-oxo-L-norvaline, L- (α -S,5S) - α -amino-3-chloro-4, 5-dihydro-5-isoxazoleacetic acid, azaserine.
4. The metabolic checkpoint-based human serum albumin nanopharmaceutical according to claim 1, wherein the indoleamine-2,3-dioxygenase inhibitor is an immunoactivator.
5. The metabolic checkpoint-based human serum albumin nanopharmaceutical according to claim 4, wherein the immune activator is 1-methyltryptophan.
6. The metabolic checkpoint-based human serum albumin nanopharmaceutical according to claim 1, wherein the human serum albumin encapsulates the prodrug molecular core by hydrophobic interactions.
7. The metabolic checkpoint-based human serum albumin nanopharmaceutical according to claim 1, wherein the prodrug molecular core has the structure of formula (I):
wherein n is any one of 0, 1, 2,3, 4,5, 6 and 7; r1And R2Selected from H or alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl; r3Is H or can be adjacent to R3The nitrogen forms an amide bond, a carbamate bond, a phosphoramidate bond, a phosphorodiamidite bond; r3' is selected from H or C1-C8 alkyl, substituted C1-C8 alkyl, or R3And R3Together,' form a ring structure comprising-C (═ O) -G-C (═ O) -, where G is selected from the group consisting of: C1-C8 alkylene, C1-C8 heteroalkylene, C5-C8 cycloalkylene, C6-C12 arylene, C5-C12 heteroarylene, divalent C4-C10 heterocycle, each of which may be optionally substituted.
8. The method for preparing a human serum albumin nano-drug based on a metabolic checkpoint as claimed in claim 1, comprising the steps of:
(1) synthesis of 9-fluorenylmethoxycarbonyl protected (indoleamine-2,3-dioxygenase inhibitor) -metabolic reprogramming agents;
(2) removal of the 9-fluorenylmethoxy carbonyl group to produce a (indoleamine-2,3-dioxygenase inhibitor) -metabolic reprogramming agent prodrug molecule;
(3) preparation of (indoleamine-2,3-dioxygenase inhibitor) -metabolism reprogramming agent human serum albumin nano-drug.
9. The preparation method according to claim 8, wherein in the step (1), the isopropyl-metabolic reprogramming agent and the 9-fluorenylmethoxycarbonyl-indoleamine-2, 3-dioxygenase inhibitor are coupled to generate the 9-fluorenylmethoxycarbonyl-protected indoleamine-2,3-dioxygenase inhibitor-metabolic reprogramming agent in the presence of benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate and diisopropylethylamine.
10. The preparation method of claim 8, wherein in the step (2), the product obtained in the step (1) is reacted overnight under the action of piperidine to remove the carbonyl group of 9-fluorenylmethoxy group, so as to obtain the prodrug molecule.
11. The preparation method of claim 8, wherein in the step (3) (1), the prodrug molecule obtained in the step (2) is dissolved in an organic solvent, and the prodrug molecule solution is slowly dripped into an aqueous solution of human serum albumin under the continuous action of external force to induce self-assembly of human serum albumin molecules, and the human serum albumin nano-drug based on the metabolic checkpoint is obtained through purification and filtration.
12. Use of the metabolism checkpoint based human serum albumin nano-drug according to any one of claims 1 to 7 in tumor immunotherapy.
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| CN110585131A (en) * | 2019-09-20 | 2019-12-20 | 宁夏医科大学 | Chemotherapy drug co-loaded 1-methyltryptophan immune prodrug micelle, preparation method and application thereof |
| CN112920092A (en) * | 2015-07-31 | 2021-06-08 | 约翰霍普金斯大学 | Prodrugs of glutamine analogs |
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| EP3923934A4 (en) * | 2019-02-11 | 2022-11-16 | Dracen Pharmaceuticals, Inc. | Method of preparing a don prodrug from l-pyroglutamic acid |
| WO2022232565A1 (en) * | 2021-04-29 | 2022-11-03 | The Johns Hopkins University | Prodrugs of 6-diazo-5-oxo-l-norleucine |
| CN114306620A (en) * | 2021-12-07 | 2022-04-12 | 深圳先进技术研究院 | Human serum albumin nano-drug based on metabolism check point and preparation method and application thereof |
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| CN112920092A (en) * | 2015-07-31 | 2021-06-08 | 约翰霍普金斯大学 | Prodrugs of glutamine analogs |
| CN108295046A (en) * | 2016-12-30 | 2018-07-20 | 中国科学院深圳先进技术研究院 | The preparation method and albumin nanoparticle obtained of a kind of albumin nanoparticle and application |
| CN110585131A (en) * | 2019-09-20 | 2019-12-20 | 宁夏医科大学 | Chemotherapy drug co-loaded 1-methyltryptophan immune prodrug micelle, preparation method and application thereof |
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| WO2023104021A1 (en) * | 2021-12-07 | 2023-06-15 | 深圳先进技术研究院 | Metabolic checkpoint-based human serum albumin nano-drug, and preparation method therefor and application thereof |
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